1
00:00:06,397 --> 00:00:07,698
>> GOOD AFTERNOON.

2
00:00:07,698 --> 00:00:12,737
THANKS FOR COMING IN PERSON. 

3
00:00:12,737 --> 00:00:17,642
IT'S A PLEASURE TO INTRODUCE

4
00:00:17,642 --> 00:00:19,977
JAKOB, UNDERGRADUATE AT

5
00:00:19,977 --> 00:00:23,881
DARTMOUTH, UC-BERKELEY FOR

6
00:00:23,881 --> 00:00:34,425
GRADUATE STUDIES, DEVELOPED THE

7
00:00:36,227 --> 00:00:41,866
SYSTEM DELIVERING FLAGELLAN INTO

8
00:00:41,866 --> 00:00:44,435
THE CYTOSOL AND SHOWING THEY ARE

9
00:00:44,435 --> 00:00:49,540
AN IMPORTANT OUTPIT OF

10
00:00:49,540 --> 00:00:50,608
ACTIVATION.

11
00:00:50,608 --> 00:00:54,879
HE WENT TO UCSF, POSTDOC WORK

12
00:00:54,879 --> 00:00:57,415
WITH RICH LOCKSLY, PUBLISHED

13
00:00:57,415 --> 00:01:02,653
WHAT'S A SEMINAL STUDY

14
00:01:02,653 --> 00:01:03,854
IDENTIFYING MURKY CHEMOSENSORY

15
00:01:03,854 --> 00:01:05,256
EPITHELIAL CELLS AS DOMINANT

16
00:01:05,256 --> 00:01:07,592
SOURCE OF INTERLEUKIN 25 THAT

17
00:01:07,592 --> 00:01:09,794
ACTIVATES TYPE 2 INNATE LYMPHOID

18
00:01:09,794 --> 00:01:12,029
CELLS IN THE INTESTINE, THE

19
00:01:12,029 --> 00:01:13,898
FIRST LINK BETWEEN INTESTINAL

20
00:01:13,898 --> 00:01:16,233
SPECIFIC CELL TYPE AND

21
00:01:16,233 --> 00:01:17,301
ACTIVATION OF TYPE 2

22
00:01:17,301 --> 00:01:17,868
INFLAMMATION.

23
00:01:17,868 --> 00:01:18,936
FROM THERE HE STARTED

24
00:01:18,936 --> 00:01:20,838
INDEPENDENT LAB AT UNIVERSITY OF

25
00:01:20,838 --> 00:01:24,208
WASHINGTON IN SEATTLE,

26
00:01:24,208 --> 00:01:26,077
CONTINUING TO STUDY INITIATORS

27
00:01:26,077 --> 00:01:27,545
OF TYPE 2 INFLAMMATION,

28
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INDEPENDENT GROUP MADE ONE OF

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00:01:28,813 --> 00:01:29,413
THE -- ANOTHER IMPORTANT

30
00:01:29,413 --> 00:01:31,616
CONTRIBUTION TO TRY TO

31
00:01:31,616 --> 00:01:33,851
UNDERSTAND HOW THE TUFTS CELLS

32
00:01:33,851 --> 00:01:37,788
SENSE EXOGENOUS STIMULI AND

33
00:01:37,788 --> 00:01:41,125
EXTENDING FROM OUR CANONICAL

34
00:01:41,125 --> 00:01:45,863
RECOGNITION PARADIGM THE SHOW

35
00:01:45,863 --> 00:01:48,633
METABOLITE IS A TRIGGER FOR CELL

36
00:01:48,633 --> 00:01:49,834
ACTIVATION THAT TRIGGERS TYPE 2

37
00:01:49,834 --> 00:01:51,836
INFLAMMATION, CONTINUING TO

38
00:01:51,836 --> 00:01:53,704
STUDY IMPORTANT DETAILS IN THIS

39
00:01:53,704 --> 00:01:55,773
BIOLOGY, REALLY EMERGED AS THE

40
00:01:55,773 --> 00:01:57,508
KEY FIGURE IN OUR UNDERSTANDING

41
00:01:57,508 --> 00:01:58,976
OF TUFT CELL TYPE 2 INTERFACE,

42
00:01:58,976 --> 00:02:04,949
WE'RE IN FOR AN INTERESTING

43
00:02:04,949 --> 00:02:06,150
TALK.

44
00:02:06,150 --> 00:02:07,184
WELCOME, JAKOB.

45
00:02:07,184 --> 00:02:07,451
[APPLAUSE]

46
00:02:07,451 --> 00:02:07,852
>> OKAY.

47
00:02:07,852 --> 00:02:09,754
THANK YOU VERY MUCH FOR THE

48
00:02:09,754 --> 00:02:10,554
INTRODUCTION.

49
00:02:10,554 --> 00:02:11,355
THANK YOU FOR COMING.

50
00:02:11,355 --> 00:02:13,958
THIS IS MY FIRST TIME ON THE NIH

51
00:02:13,958 --> 00:02:15,059
CAMPUS, REALLY EXCITING.

52
00:02:15,059 --> 00:02:17,028
IT'S BEEN A FUN DAY.

53
00:02:17,028 --> 00:02:19,563
AND FEELS LIKE COMING TO THE

54
00:02:19,563 --> 00:02:20,398
MOTHER SHIP.

55
00:02:20,398 --> 00:02:25,436
SO I'M PLEASED TO BE HERE.

56
00:02:25,436 --> 00:02:28,205
I'M GOING TO TALK PRETTY MUCH

57
00:02:28,205 --> 00:02:29,306
ENTIRELY ABOUT TUFT CELLS, WE

58
00:02:29,306 --> 00:02:30,608
HAVE PROJECTS DEVELOPING IN THE

59
00:02:30,608 --> 00:02:34,111
LAB NOW THAT ARE HEADING IN NEW

60
00:02:34,111 --> 00:02:37,014
DIRECTIONS, AND REALLY LET ME

61
00:02:37,014 --> 00:02:38,582
JUST SAY INITIALLY THAT THE

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00:02:38,582 --> 00:02:41,018
THEME OF THE LAB AND UNIFYING

63
00:02:41,018 --> 00:02:43,954
INTEREST IS TYPE 2 IMMUNITY, THE

64
00:02:43,954 --> 00:02:46,023
IMMUNE RESPONSE GENERATED BY

65
00:02:46,023 --> 00:02:52,997
PARASITIC WORMS AND ALLERGYINS.

66
00:02:52,997 --> 00:02:55,666
THERE HAVE BEEN DECADES OF

67
00:02:55,666 --> 00:02:57,068
SEMINAL STUDIES TO UNDERSTAND

68
00:02:57,068 --> 00:02:58,302
TYPE 2 IMMUNITY.

69
00:02:58,302 --> 00:03:01,172
ANYBODY WHO SUFFERS FROM

70
00:03:01,172 --> 00:03:02,473
ALLERGIES KNOWS CLINICAL

71
00:03:02,473 --> 00:03:04,775
MANIFESTATIONS AND WE KNOW A LOT

72
00:03:04,775 --> 00:03:11,115
OF CELLS DRIVING THAT, ADAPTIVE

73
00:03:11,115 --> 00:03:12,783
CELLS, MORE RECENTLY THERE WAS

74
00:03:12,783 --> 00:03:15,019
ADDITION OF THESE INNATE

75
00:03:15,019 --> 00:03:15,786
LYMPHOID CELLS DISTINGUISHED BY

76
00:03:15,786 --> 00:03:18,022
BEING RESIDENT IN THE TISSUE AND

77
00:03:18,022 --> 00:03:19,757
THEREFORE READY TO RESPOND

78
00:03:19,757 --> 00:03:20,357
ANYTIME.

79
00:03:20,357 --> 00:03:22,059
AND WE KNOW THE DOMINANT

80
00:03:22,059 --> 00:03:23,627
CYTOKINES THAT ARE DRIVING ALL

81
00:03:23,627 --> 00:03:25,463
OF THIS.

82
00:03:25,463 --> 00:03:26,997
BUT WHERE A LOT OF LACK OF

83
00:03:26,997 --> 00:03:29,233
UNDERSTANDING REMAINS STILL IS

84
00:03:29,233 --> 00:03:36,373
HOW YOU GET FROM THESE STIMULI,

85
00:03:36,373 --> 00:03:39,009
DIVERSE SET OF STIMULI, TO

86
00:03:39,009 --> 00:03:40,144
ACTIVATING CELLS IN THE

87
00:03:40,144 --> 00:03:40,544
UNDERLYING TISSUE.

88
00:03:40,544 --> 00:03:43,814
A LOT OF TIMES THESE STIMULI ARE

89
00:03:43,814 --> 00:03:45,716
NOT INVASIVE, HOW DO YOU

90
00:03:45,716 --> 00:03:47,418
TRANSMIT SIGNAL FROM THE OUTSIDE

91
00:03:47,418 --> 00:03:47,985
WORLD TO INSIDE WORLD?

92
00:03:47,985 --> 00:03:51,322
THE BARRIER OF COURSE IS A KEY

93
00:03:51,322 --> 00:03:53,824
PART, EPITHELIUM, AND WHAT WE'VE

94
00:03:53,824 --> 00:03:56,127
FOCUSED ON IS THIS ROLE OF

95
00:03:56,127 --> 00:03:59,463
EPITHELIAL TUFT CELLS IN THIS

96
00:03:59,463 --> 00:03:59,697
PROCESS.

97
00:03:59,697 --> 00:04:01,065
SO, LET ME SAY -- TELL WHERE YOU

98
00:04:01,065 --> 00:04:01,932
WE'RE GOING TODAY.

99
00:04:01,932 --> 00:04:03,567
FIRST THING I THINK AT THIS

100
00:04:03,567 --> 00:04:07,104
POINT PEOPLE HAVE MAYBE HEARD OF

101
00:04:07,104 --> 00:04:08,839
TUFT CELLS BUT THEY ARE STILL --

102
00:04:08,839 --> 00:04:09,874
I FIND PEOPLE DON'T NECESSARILY

103
00:04:09,874 --> 00:04:11,342
KNOW THE HISTORY THAT WELL.

104
00:04:11,342 --> 00:04:13,644
WE'LL TAKE TIME TO TELL YOU THE

105
00:04:13,644 --> 00:04:15,079
BACKGROUND, BOTH OF OTHER

106
00:04:15,079 --> 00:04:16,480
PEOPLE'S WORK AND WHERE WE SORT

107
00:04:16,480 --> 00:04:18,883
OF HAVE TAKEN OVER AND WHAT WE

108
00:04:18,883 --> 00:04:19,350
FOUND.

109
00:04:19,350 --> 00:04:21,685
AND THEN I'LL TELL YOU ONE

110
00:04:21,685 --> 00:04:23,087
RECENTLY PUBLISHED STORY THAT

111
00:04:23,087 --> 00:04:25,156
HIGHLIGHTS THE WAY TUFT CELLS

112
00:04:25,156 --> 00:04:27,124
INTERACT WITH THE EPITHELIUM TO

113
00:04:27,124 --> 00:04:28,225
REGULATE ITS PHYSIOLOGY AND HOW

114
00:04:28,225 --> 00:04:30,494
THAT LINKS WITH TYPE 2 IMMUNITY.

115
00:04:30,494 --> 00:04:33,397
AND THEN UNPUBLISHED STUDY WHERE

116
00:04:33,397 --> 00:04:34,465
WE'RE SORT OF EXTENDING THE TIME

117
00:04:34,465 --> 00:04:36,400
SCALE IN WHICH WE THINK ABOUT

118
00:04:36,400 --> 00:04:37,868
THIS, TO THINK ABOUT CHRONIC AND

119
00:04:37,868 --> 00:04:40,337
ADAPT 

120
00:04:40,337 --> 00:04:43,207
ADAPTIVE RESPONSES AND ROLE OF

121
00:04:43,207 --> 00:04:46,811
TUFT CELLS ESPECIALLY

122
00:04:46,811 --> 00:04:48,112
INTERSECTING WITH Th2s.

123
00:04:48,112 --> 00:04:49,780
TUFT CELLS HAVE BEEN KNOWN ABOUT

124
00:04:49,780 --> 00:04:52,416
FOR MORE THAN 60 YEARS BECAUSE

125
00:04:52,416 --> 00:04:54,185
WHEN PEOPLE STARTED TAKING

126
00:04:54,185 --> 00:04:56,554
ELECTRON MICROSCOPY IMAGES, THEY

127
00:04:56,554 --> 00:04:58,689
IMMEDIATELY SAW THAT EVERY ONCE

128
00:04:58,689 --> 00:05:02,026
IN A WHILE, RARELY, THEY WOULD

129
00:05:02,026 --> 00:05:03,828
FIND A CELL THAT LOOKED

130
00:05:03,828 --> 00:05:06,130
DIFFERENT, DEFINED BY A TUFT AT

131
00:05:06,130 --> 00:05:06,797
THE TOP.

132
00:05:06,797 --> 00:05:08,799
SCANNING E.M. SHOWS I THE WAY

133
00:05:08,799 --> 00:05:11,035
THE TUFT OF THE TUFT CELL

134
00:05:11,035 --> 00:05:12,102
CONVERGES UP OUT OF THE

135
00:05:12,102 --> 00:05:13,470
EPITHELIAL BARRIER AND REACHES

136
00:05:13,470 --> 00:05:15,940
INTO THE LUMEN OF THE TISSUE

137
00:05:15,940 --> 00:05:16,340
THAT IT'S IN.

138
00:05:16,340 --> 00:05:18,275
AND SO THERE WAS REALLY

139
00:05:18,275 --> 00:05:19,476
BEAUTIFUL EARLY E.M. WORK THAT

140
00:05:19,476 --> 00:05:21,478
LOOKED AT THE CELL BIOLOGY OF

141
00:05:21,478 --> 00:05:24,415
THESE THINGS, A LOT OF

142
00:05:24,415 --> 00:05:25,049
PHENOTYPING, JUST IDENTIFYING

143
00:05:25,049 --> 00:05:25,850
WHERE THEY WERE.

144
00:05:25,850 --> 00:05:26,917
THEY FOUND THEY WERE RARE CELLS

145
00:05:26,917 --> 00:05:30,187
AND THEY WERE IN LOTS OF MUCOSAL

146
00:05:30,187 --> 00:05:32,790
BARRIERS, UP AND DOWN THE

147
00:05:32,790 --> 00:05:36,327
INTESTINAL TRACT, PARTS OF

148
00:05:36,327 --> 00:05:37,528
AIRWAY, TRACKA, NASAL PASSAGE

149
00:05:37,528 --> 00:05:39,530
BUT NOT THE LUNG.

150
00:05:39,530 --> 00:05:40,698
INTERESTINGLY IN THE THYMUS.

151
00:05:40,698 --> 00:05:42,132
THE THING LACKING WAS ANY

152
00:05:42,132 --> 00:05:44,301
UNDERSTANDING OF FUNCTION.

153
00:05:44,301 --> 00:05:46,103
THIS WAS JUST NOTICING THERE ARE

154
00:05:46,103 --> 00:05:47,004
UNUSUAL CELLS.

155
00:05:47,004 --> 00:05:48,772
AND THAT STARTED TO CHANGE IN

156
00:05:48,772 --> 00:05:50,274
THE LATE '90s, EARLY 2000s,

157
00:05:50,274 --> 00:05:52,710
REALLY BECAUSE OF PEOPLE WHO

158
00:05:52,710 --> 00:05:53,277
WERE STUDYING TASTE

159
00:05:53,277 --> 00:05:54,211
TRANSDUCTION, AND THEY REALIZE

160
00:05:54,211 --> 00:05:56,480
THE THE TOOLS THEY WERE BUILDING

161
00:05:56,480 --> 00:05:58,249
WERE LIKE THE ANTIBODIES AND

162
00:05:58,249 --> 00:06:00,584
REPORTER MICE WERE ALSO MARKING

163
00:06:00,584 --> 00:06:01,952
THESE RARE CELLS IN THE

164
00:06:01,952 --> 00:06:04,922
INTESTINE, AND IN OTHER PLACES,

165
00:06:04,922 --> 00:06:05,756
THESE TUFT CELLS.

166
00:06:05,756 --> 00:06:08,759
AND SO ONCE WE HAD THE FIRST

167
00:06:08,759 --> 00:06:13,831
TRANSCRIPTOME FOR TUFT IT BECAME

168
00:06:13,831 --> 00:06:14,965
CLEAR THE SHARED PROGRAM BETWEEN

169
00:06:14,965 --> 00:06:16,800
TYPE 2 TASTE RECEPTOR CELLS AND

170
00:06:16,800 --> 00:06:18,702
BETWEEN THE TUFT CELLS, AND THIS

171
00:06:18,702 --> 00:06:20,971
IS PARTICULARLY TRUE FOR KIND OF

172
00:06:20,971 --> 00:06:22,873
THE INTERNAL WORKINGS OF THE

173
00:06:22,873 --> 00:06:24,642
CELL AND TRANSDUCTION PATHWAYS.

174
00:06:24,642 --> 00:06:30,748
SO THEY BOTH SHARE THESE

175
00:06:30,748 --> 00:06:32,917
SPECIALIZED SIGNAL THROUGH PLC

176
00:06:32,917 --> 00:06:35,486
BETA2, MOST IMPORTANTLY THIS

177
00:06:35,486 --> 00:06:38,689
TRPM 5 CHANNEL, CALCIUM GATED

178
00:06:38,689 --> 00:06:39,957
SODIUM CHANNEL THAT DEPOLARIZES

179
00:06:39,957 --> 00:06:41,258
CELLS, HOW TASTE SIGNALING

180
00:06:41,258 --> 00:06:42,927
THROUGH RECEPTORS IS CONVERTED

181
00:06:42,927 --> 00:06:45,129
INTO A NEURONAL SIGNAL DOWN

182
00:06:45,129 --> 00:06:45,329
BELOW.

183
00:06:45,329 --> 00:06:48,632
SO TUFT CELLS HAVE THESE THINGS.

184
00:06:48,632 --> 00:06:51,435
TURNS OUT ALSO THERE'S THAT

185
00:06:51,435 --> 00:06:53,237
MASTER TRANSCRIPTION FACTOR FOR

186
00:06:53,237 --> 00:06:55,439
THE TYPE 2 LINEAGE AND FOR TUFT

187
00:06:55,439 --> 00:06:57,942
CELLS SO MICE LACKING THIS DON'T

188
00:06:57,942 --> 00:06:59,443
HAVE ANY TUFT CELLS.

189
00:06:59,443 --> 00:07:01,679
THE FIRST KIND OF UNDERSTANDINGS

190
00:07:01,679 --> 00:07:02,947
OF FUNCTION EMENTIONED SOON

191
00:07:02,947 --> 00:07:04,715
AFTER THAT, AND THIS WAS WORK

192
00:07:04,715 --> 00:07:06,583
DONE IN THE AIRWAYS, RECOGNIZING

193
00:07:06,583 --> 00:07:10,521
TUFT CELLS COULD BE ACTIVATED BY

194
00:07:10,521 --> 00:07:12,356
SOME BACTERIA DERIVED LIGANDS

195
00:07:12,356 --> 00:07:18,796
LIKE SENSING MOLECULES, FOR

196
00:07:18,796 --> 00:07:20,064
EXAMPLE, THROUGH CANONICAL TASTE

197
00:07:20,064 --> 00:07:21,265
RECEPTORS, THE FOCUS THAT HE

198
00:07:21,265 --> 00:07:27,938
COULD TALK TO NEURONS IN THE

199
00:07:27,938 --> 00:07:29,273
AIRWAYS AND INDUCE NEUROGENIC

200
00:07:29,273 --> 00:07:31,375
INFLAMMATION AND STUFF LIKE

201
00:07:31,375 --> 00:07:34,578
THAT. 

202
00:07:34,578 --> 00:07:35,179
WHAT REMAINS MYSTERIOUS WHAT

203
00:07:35,179 --> 00:07:36,146
THEY WERE DOING IN THE

204
00:07:36,146 --> 00:07:37,815
INTESTINE.

205
00:07:37,815 --> 00:07:44,021
I   STUMBLED INTO THIS AS A

206
00:07:44,021 --> 00:07:46,390
POSTDOC, WE KNOW IL-25 WAS

207
00:07:46,390 --> 00:07:49,693
IMPORTANT FOR TYPE 2 IMMUNE

208
00:07:49,693 --> 00:07:51,528
RESPONSES, ESPECIALLY IN THE

209
00:07:51,528 --> 00:07:56,734
INTESTINE, DIDN'T KNOW WHERE

210
00:07:56,734 --> 00:08:00,304
CYTOKINE WAS COMING FROM, MADE A

211
00:08:00,304 --> 00:08:01,105
REPORTER MOUSE, INSERTED

212
00:08:01,105 --> 00:08:02,706
DOWNSTREAM AND LOOKED FOR RED

213
00:08:02,706 --> 00:08:04,441
CELLS, FOUND THEM IN BARRIER

214
00:08:04,441 --> 00:08:08,579
SITES, RARE, GREEN IS MARKING

215
00:08:08,579 --> 00:08:11,248
THE EPCAM BARRIER IN THIS CASE

216
00:08:11,248 --> 00:08:12,349
OF THE DUODENUM, MAYBE ONE IN

217
00:08:12,349 --> 00:08:15,953
100 OR FEWER OF THE CELLS ARE

218
00:08:15,953 --> 00:08:16,520
IL-25 PRODUCERS.

219
00:08:16,520 --> 00:08:18,822
AND SO WHAT WE FIGURED OUT AT

220
00:08:18,822 --> 00:08:19,990
THE TIME, THOSE WERE ALL TUFT

221
00:08:19,990 --> 00:08:24,061
CELLS, EVERY ONE OF THESE IL-25

222
00:08:24,061 --> 00:08:24,428
POSITIVE CELLS.

223
00:08:24,428 --> 00:08:25,496
SO FAR WE'VE NEVER SEEN ANY

224
00:08:25,496 --> 00:08:27,931
OTHER CELL TURN RED WITH THIS

225
00:08:27,931 --> 00:08:28,565
REPORTER.

226
00:08:28,565 --> 00:08:30,100
AND CONVERSELY EVERY TUFT CELL

227
00:08:30,100 --> 00:08:34,171
AS IT WAS BORN AS PART OF THE

228
00:08:34,171 --> 00:08:38,442
PROGRAM EXPRESSED IL-25 AT LEAST

229
00:08:38,442 --> 00:08:38,909
TRANSCRIPTIONALLY.

230
00:08:38,909 --> 00:08:41,812
WE THINK MORE HAPPENS LATER.

231
00:08:41,812 --> 00:08:42,079
OKAY.

232
00:08:42,079 --> 00:08:43,580
SO, THE OTHER THING THOUGH THAT

233
00:08:43,580 --> 00:08:46,750
HAPPENS IS THAT THE NUMBERS OF

234
00:08:46,750 --> 00:08:48,152
TUFT CELLS IS ASTONISHLY DYNAMIC

235
00:08:48,152 --> 00:08:49,586
ESPECIALLY IN SMALL INTESTINE,

236
00:08:49,586 --> 00:08:51,255
UNIQUELY IN THE SMALL INTESTINE

237
00:08:51,255 --> 00:08:51,588
REALLY.

238
00:08:51,588 --> 00:08:56,627
ON THE LEFT HERE ARE TWO -- YOU

239
00:08:56,627 --> 00:08:57,795
WITH SEE TWO ENTIRE VILLI, ONLY

240
00:08:57,795 --> 00:09:00,998
THREE TUFT CELLS MARKED WITH

241
00:09:00,998 --> 00:09:02,733
THIS ANTIBODY FOR DCLK 1, SEVEN

242
00:09:02,733 --> 00:09:04,601
DAYS AFTER PUTTING A WORM IN THE

243
00:09:04,601 --> 00:09:06,570
MOUSE HAVE YOU DOZENS OF TUFT

244
00:09:06,570 --> 00:09:11,375
CELLS IN THE SINGLE VILLUS,

245
00:09:11,375 --> 00:09:12,476
MASSIVE HYPERPLASIA EMERGES

246
00:09:12,476 --> 00:09:15,446
BECAUSE OF THE DYNAMICS OF THE

247
00:09:15,446 --> 00:09:16,046
INTESTINAL EPITHELIUM.

248
00:09:16,046 --> 00:09:19,049
SO THIS IS WHAT IT LOOKED LIKE

249
00:09:19,049 --> 00:09:20,818
SCANNING E.M., LARGE VILLI IN

250
00:09:20,818 --> 00:09:22,986
THE SMALL INTESTINE, AND SMALLER

251
00:09:22,986 --> 00:09:23,921
CRYPTS DOWN BELOW.

252
00:09:23,921 --> 00:09:25,155
THE STEM CELLS RESIDE AT THE

253
00:09:25,155 --> 00:09:27,958
BASE OF THE CRYPT WITH PANNA

254
00:09:27,958 --> 00:09:30,461
CELLS, GIVE RISE TO CELLS THAT

255
00:09:30,461 --> 00:09:33,397
DIFFERENTIATE AS THEY MOVE UP,

256
00:09:33,397 --> 00:09:36,200
ADOPT INTERNAL FATES IN THE

257
00:09:36,200 --> 00:09:38,869
VILLUS, EVENTUALLY LOST.

258
00:09:38,869 --> 00:09:40,504
MOST ABSORBED AT ENTEROCYTES,

259
00:09:40,504 --> 00:09:43,373
LESS FREQUENT LINEAGES LIKE THE

260
00:09:43,373 --> 00:09:44,608
GOBLET CELLS AND HORMONE CELLS,

261
00:09:44,608 --> 00:09:47,344
AND TUFT CELLS AS WELL.

262
00:09:47,344 --> 00:09:50,714
AND THE KEY THING IS THAT THESE

263
00:09:50,714 --> 00:09:51,815
CELLS ARE SHORT-LIVED.

264
00:09:51,815 --> 00:09:53,150
FOUR TO FIVE DAYS, REPLACE

265
00:09:53,150 --> 00:09:54,818
BASICALLY THE ENTIRE LINING OF

266
00:09:54,818 --> 00:09:57,054
THE EPITHELIUM, TRUE IN RODENTS

267
00:09:57,054 --> 00:09:57,421
AND HUMANS.

268
00:09:57,421 --> 00:09:59,256
AND SO IF YOU CAN CHANGE THE

269
00:09:59,256 --> 00:10:02,359
OUTPUT FROM THIS STEM CELL

270
00:10:02,359 --> 00:10:03,560
COMPARTMENT, YOU CAN COMPLETELY

271
00:10:03,560 --> 00:10:05,829
CHANGE COMPOSITION AND FUNCTION

272
00:10:05,829 --> 00:10:07,464
OF EPITHELIUM WITHIN DAYS.

273
00:10:07,464 --> 00:10:10,534
SO THIS IS WHAT WE DESCRIBED

274
00:10:10,534 --> 00:10:11,135
TOGETHER WITH PHILIP'S LAB IN

275
00:10:11,135 --> 00:10:12,636
FRANCE AT THE TIME.

276
00:10:12,636 --> 00:10:15,072
WE CALLED IT THE TUFT IL-2

277
00:10:15,072 --> 00:10:15,806
CIRCUIT, WHAT'S HAPPENING IS

278
00:10:15,806 --> 00:10:16,907
THAT WHEN THERE ARE WORMS

279
00:10:16,907 --> 00:10:19,543
PRESENT IN THE LUMEN OF THE

280
00:10:19,543 --> 00:10:21,311
SMALL INTESTINE TUFT CELLS

281
00:10:21,311 --> 00:10:23,413
SOMEHOW RESPOND TO THEM.

282
00:10:23,413 --> 00:10:26,483
RELEASE IL-25 AND THAT ACTS ON

283
00:10:26,483 --> 00:10:34,992
THESE ILC 2s TO MAKE CANONICAL

284
00:10:34,992 --> 00:10:37,494
TYPE 2 CYTOKINES.

285
00:10:37,494 --> 00:10:38,162
THE 13 SIGNALS BACK TO SOMEWHERE

286
00:10:38,162 --> 00:10:38,829
IN THE CRYPT.

287
00:10:38,829 --> 00:10:42,032
WE DON'T FLOW IF IT'S STEM CELLS

288
00:10:42,032 --> 00:10:43,667
OR SOME TRANSIT AMPLIFYING ZONE,

289
00:10:43,667 --> 00:10:44,635
CELLS HAVE NOT FULLY DECIDED IF

290
00:10:44,635 --> 00:10:46,703
THEY ARE GOING TO BE A TUFT

291
00:10:46,703 --> 00:10:49,039
CELL, IT CAN EXPAND THIS TUFT

292
00:10:49,039 --> 00:10:51,542
CELL LINEAGE AND SO ONCE YOU'VE

293
00:10:51,542 --> 00:10:54,478
TURNED ON IL-15 YOU GET TUFT

294
00:10:54,478 --> 00:10:59,416
CELL HYPERPLASIA, A FEED-FORWARD

295
00:10:59,416 --> 00:11:00,918
TUFT IL-CIRCUIT.

296
00:11:00,918 --> 00:11:04,154
IT FELT LIKE THERE ARE MISSING

297
00:11:04,154 --> 00:11:06,623
LINKS, SIT AT THE BARRIER,

298
00:11:06,623 --> 00:11:09,493
EQUIPPED WITH THINGS TO TALK TO

299
00:11:09,493 --> 00:11:10,427
THE UNDERLYING TISSUE.

300
00:11:10,427 --> 00:11:14,097
RIGHT AT THAT SAME TIME MICHAEL

301
00:11:14,097 --> 00:11:15,432
AND WENDY'S LAB ARRIVED AT A

302
00:11:15,432 --> 00:11:18,702
SIMILAR FINDING FROM A DIFFERENT

303
00:11:18,702 --> 00:11:19,736
APPROACH, MICROBIOME LAB,

304
00:11:19,736 --> 00:11:22,306
REALIZED NUMBERS OF ACTIVITY

305
00:11:22,306 --> 00:11:25,342
CELLS WERE DIFFERENT IN IN-HOUSE

306
00:11:25,342 --> 00:11:29,780
MICE AND JAK MICE AND TRACED

307
00:11:29,780 --> 00:11:34,284
BACK TO TRICHOMONAS, COMMON IN

308
00:11:34,284 --> 00:11:35,652
VIVARIUMS, AND ACTIVATE THE SAME

309
00:11:35,652 --> 00:11:36,720
CIRCUIT DESCRIBED BEAUTIFULLY.

310
00:11:36,720 --> 00:11:39,289
AND SO MICE LACKING ANY

311
00:11:39,289 --> 00:11:40,190
COMPONENT OF THE CIRCUIT, EITHER

312
00:11:40,190 --> 00:11:43,327
DON'T CLEAR WORMS OR DON'T SENSE

313
00:11:43,327 --> 00:11:43,627
THE PROTUS.

314
00:11:43,627 --> 00:11:48,832
IF YOU GET RID OF TUFT CELLS OR

315
00:11:48,832 --> 00:11:51,535
IL-25, FOR EXAMPLE.

316
00:11:51,535 --> 00:11:54,438
SO, WE'VE BEEN TRYING TO FILL IN

317
00:11:54,438 --> 00:11:56,206
THESE GAPS AND UNDERSTAND HOW IS

318
00:11:56,206 --> 00:12:00,244
THE SENSING WORKING, HOW ARE

319
00:12:00,244 --> 00:12:01,511
PROTEUS AND WORMS DETECTION.

320
00:12:01,511 --> 00:12:03,880
THE WORM DETECTION THAT WE DO

321
00:12:03,880 --> 00:12:06,016
NOT UNDERSTAND YET REMAINS A

322
00:12:06,016 --> 00:12:07,484
BIGGEST QUESTION, AND WHAT ELSE

323
00:12:07,484 --> 00:12:11,355
ARE THEY DOING BESIDES MAKING

324
00:12:11,355 --> 00:12:16,226
IL-25 AND HOW DOES SIGNALING AND

325
00:12:16,226 --> 00:12:16,994
DIFFERENTIATION WORK.

326
00:12:16,994 --> 00:12:20,864
ONE OF OUR FIRST IN SIGHTS WAS

327
00:12:20,864 --> 00:12:23,834
SENSING OF SUCCINATE BY THE

328
00:12:23,834 --> 00:12:25,669
FIRST TWO GRAD STUDENTS IN THE

329
00:12:25,669 --> 00:12:28,538
LAB, TUFT CELLS ARE EQUIPPED

330
00:12:28,538 --> 00:12:31,174
WITH SUCCINATE RECEPTORS THAT

331
00:12:31,174 --> 00:12:32,075
SENSES EXTRACELLULAR SUCCINATE,

332
00:12:32,075 --> 00:12:34,177
AND THAT CAN INDUCE CALCIUM FLUX

333
00:12:34,177 --> 00:12:36,647
THAT WILL OPEN TRPM 5, AND SO IT

334
00:12:36,647 --> 00:12:37,848
TURNS OUT IF YOU PUT SUCCINATE

335
00:12:37,848 --> 00:12:39,883
IN THE DRINKING WATER OF THE

336
00:12:39,883 --> 00:12:42,286
MICE, THAT SINGLE MOLECULE IS

337
00:12:42,286 --> 00:12:47,457
SUSPICIOUS TO  TURN ON THE ENTIE

338
00:12:47,457 --> 00:12:48,458
CIRCUIT, THE MOUSE LOOKS LIKE

339
00:12:48,458 --> 00:12:51,862
IT'S INFECTED WITH A  WORM.

340
00:12:51,862 --> 00:12:55,165
THIS PATHWAY IS REQUIRED FOR

341
00:12:55,165 --> 00:12:56,867
SENSING OF THESE, WHICH MAKE A

342
00:12:56,867 --> 00:12:58,802
LOT OF SUCCINATE UNDER ANAEROBIC

343
00:12:58,802 --> 00:13:00,103
CONDITIONS, DISPENSABLE FOR THE

344
00:13:00,103 --> 00:13:03,774
SENSING OF THE WORMS.

345
00:13:03,774 --> 00:13:04,841
WE DON'T UNDERSTAND THAT

346
00:13:04,841 --> 00:13:05,842
DICHOTOMY REALLY.

347
00:13:05,842 --> 00:13:09,379
NOW OTHERS ALSO HAVE SHOWN THERE

348
00:13:09,379 --> 00:13:12,082
ARE CIRCUMSTANCES OF BACTERIAL

349
00:13:12,082 --> 00:13:14,251
DYSBIOSIS, ELEVATED SUCCINATE IS

350
00:13:14,251 --> 00:13:20,791
SENSED BY TUFT CELLS AND TURNS

351
00:13:20,791 --> 00:13:21,291
ON THIS CIRCUIT.

352
00:13:21,291 --> 00:13:24,561
THIS WAS JACK'S SECOND PROJECT

353
00:13:24,561 --> 00:13:25,796
IN THE LAB.

354
00:13:25,796 --> 00:13:31,101
IN I ADDITION TO MAKING IL-25 ON

355
00:13:31,101 --> 00:13:33,804
DEMAND SYNTHESIZE AND RELEASE

356
00:13:33,804 --> 00:13:34,604
LIPID MEDIATORS, LTC 4

357
00:13:34,604 --> 00:13:39,409
SPECIFICALLY, AND HE SHOWED HOW

358
00:13:39,409 --> 00:13:42,012
THAT COOPERATES WITH IL-25 TO

359
00:13:42,012 --> 00:13:43,380
RAPIDLY INDUCE CYTOKINE

360
00:13:43,380 --> 00:13:44,614
PRODUCTION.

361
00:13:44,614 --> 00:13:45,449
INTERESTINGLY THIS IS REQUIRED

362
00:13:45,449 --> 00:13:48,685
WHEN THERE ARE WORMS,

363
00:13:48,685 --> 00:13:53,924
DISPENSABLE WHEN A PROSIOUS,  P-

364
00:13:53,924 --> 00:13:56,026
PROTEUS, A SPLIT IN THE

365
00:13:56,026 --> 00:13:56,293
PATHWAYS.

366
00:13:56,293 --> 00:13:58,161
THOSE ARE THE GAPS WE'VE FILLED

367
00:13:58,161 --> 00:13:58,962
IN.

368
00:13:58,962 --> 00:14:00,897
YOU MAY WONDER, WELL, THAT'S

369
00:14:00,897 --> 00:14:02,332
NICE IN THESE MICE BUT DOES THIS

370
00:14:02,332 --> 00:14:04,835
HAVE ANYTHING TO DO WITH HUMANS?

371
00:14:04,835 --> 00:14:07,170
I MEAN, THIS IS STILL VERY EARLY

372
00:14:07,170 --> 00:14:09,606
DAYS, UNTIL RECENTLY JUST KNEW

373
00:14:09,606 --> 00:14:11,808
THERE WERE TUFT CELLS IN HUMANS,

374
00:14:11,808 --> 00:14:13,543
GOT SINGLE CELL DATA.

375
00:14:13,543 --> 00:14:16,213
A NUMBER OF DATASETS NOW WHICH

376
00:14:16,213 --> 00:14:17,948
SATISFYINGLY SHOW THAT HUMAN

377
00:14:17,948 --> 00:14:19,616
TUFT CELLS EXPRESS BASICALLY THE

378
00:14:19,616 --> 00:14:21,218
SAME PROGRAMS, FOR EXAMPLE YOU

379
00:14:21,218 --> 00:14:22,619
CAN SEE THE SUCCINATE RECEPTOR

380
00:14:22,619 --> 00:14:24,321
HIGHER IN THE SMALL INTESTINE

381
00:14:24,321 --> 00:14:27,791
THAN COLON, SAME IN MICE.

382
00:14:27,791 --> 00:14:32,329
TRPM 5 IS REALLY HIGH, GNAT 3,

383
00:14:32,329 --> 00:14:35,799
AND THEN COMPONENTS OF THE

384
00:14:35,799 --> 00:14:36,466
SYNTHESIS PATHWAY.

385
00:14:36,466 --> 00:14:38,969
SEEMS TO BE VERY SHARED

386
00:14:38,969 --> 00:14:40,404
TRANSCRIPTIONAL PROGRAMS AND

387
00:14:40,404 --> 00:14:41,905
THEN A REALLY IMPORTANT

388
00:14:41,905 --> 00:14:45,509
CONTRIBUTION JUST LAST YEAR

389
00:14:45,509 --> 00:14:48,578
SHOWING HYPERPLASIA CAN ALSO

390
00:14:48,578 --> 00:14:52,516
OCCUR IN HUMANS, THIS REQUIRED

391
00:14:52,516 --> 00:14:54,584
OPTIMIZING ORGANOID CULTURE

392
00:14:54,584 --> 00:15:01,291
CONDITIONS TO GROW AND

393
00:15:01,291 --> 00:15:03,660
DIFFERENTIATE TUFT CELLS IN

394
00:15:03,660 --> 00:15:05,796
HUMAN ORGANOIDS, YOU GET THIS

395
00:15:05,796 --> 00:15:07,397
NICE HYPERPLASIA OF TUFT CELLS,

396
00:15:07,397 --> 00:15:11,001
SIMILAR MECHANISM IN MICE AND

397
00:15:11,001 --> 00:15:12,335
HUMANS.

398
00:15:12,335 --> 00:15:12,836
OKAY.

399
00:15:12,836 --> 00:15:13,904
SO, THAT'S MY VERY QUICK

400
00:15:13,904 --> 00:15:16,973
BACKGROUND ON TUFT CELLS AND

401
00:15:16,973 --> 00:15:19,543
WHERE WE'VE GOTTEN SO FAR.

402
00:15:19,543 --> 00:15:22,179
I'M GOING TO TALK ABOUT THIS

403
00:15:22,179 --> 00:15:23,046
PROJECT ABOUT EPITHELIAL

404
00:15:23,046 --> 00:15:24,080
PHYSIOLOGY AND TUFT CELLS.

405
00:15:24,080 --> 00:15:28,084
THIS CAME OUT OF A SIMPLE

406
00:15:28,084 --> 00:15:30,253
QUESTION WHICH WAS WHY TUFT CELL

407
00:15:30,253 --> 00:15:30,987
HYPERPLASIA HAPPENS.

408
00:15:30,987 --> 00:15:34,925
THE THINGS WE KNEW THEY HAD THIS

409
00:15:34,925 --> 00:15:36,026
SENTINEL FUNCTION, DETECTING

410
00:15:36,026 --> 00:15:37,828
SOMETHING, EARLY ACTIVATORS.

411
00:15:37,828 --> 00:15:39,062
WHY BOTHER MAKING MORE SENTINELS

412
00:15:39,062 --> 00:15:42,199
IF YOU ALREADY DETECTED WHAT YOU

413
00:15:42,199 --> 00:15:43,433
WANT TO DETECT?

414
00:15:43,433 --> 00:15:45,202
AND WE THINK WE HAVE MAYBE

415
00:15:45,202 --> 00:15:46,303
GETTING TOWARDS SOME

416
00:15:46,303 --> 00:15:48,238
UNDERSTANDING OF THIS FROM WORK

417
00:15:48,238 --> 00:15:54,244
DONE BY ANOTHER GRADUATE

418
00:15:54,244 --> 00:16:03,019
STUDENT, TYLER, BILL UP AND

419
00:16:03,019 --> 00:16:04,087
INTEREST IN ACETYLCHOLINE.

420
00:16:04,087 --> 00:16:07,824
THEY ARE THE ONLY CELLS IN THE

421
00:16:07,824 --> 00:16:09,793
INTESTINE THAT EXPRESS CHAT.

422
00:16:09,793 --> 00:16:13,697
TYLER WANTED TO KNOW WHAT THAT

423
00:16:13,697 --> 00:16:16,633
WAS FOR.

424
00:16:16,633 --> 00:16:18,635
SO, ACETYLCHOLINE IS ASSOCIATED

425
00:16:18,635 --> 00:16:20,237
WITH BEING NEUROTRANSMITTER, AND

426
00:16:20,237 --> 00:16:22,772
SO PEOPLE OF IT MADE BY NEURONS

427
00:16:22,772 --> 00:16:23,907
OR SIGNALING ON NEURONS, AND

428
00:16:23,907 --> 00:16:28,178
CERTAINLY AGAIN IN THE AIR WAIST

429
00:16:28,178 --> 00:16:29,779
THERE -- AIRWAYS THEY TALK TO

430
00:16:29,779 --> 00:16:31,014
NEURONS, COULD BE TRUE IN THE

431
00:16:31,014 --> 00:16:32,516
INTESTINE, BUT NOT REALLY WHAT

432
00:16:32,516 --> 00:16:36,152
WE FOCUSED ON ANY PARTICULAR

433
00:16:36,152 --> 00:16:36,653
PROJECT.

434
00:16:36,653 --> 00:16:37,687
INSTEAD, WE STARTED THINKING

435
00:16:37,687 --> 00:16:39,589
ABOUT ANOTHER IMPORTANT FUNCTION

436
00:16:39,589 --> 00:16:49,032
FOR ACETYLCHOLINE IN THE

437
00:16:49,032 --> 00:16:53,737
INTESTINE, IT DRIVES SECRETION

438
00:16:53,737 --> 00:16:55,038
OF MUCUS, ANTIMICROBIAL

439
00:16:55,038 --> 00:16:58,608
PEPTIDES, FLUID OR CHLORIDE IONS

440
00:16:58,608 --> 00:17:04,314
THAT THEN PULL FLUID OUT OF THE

441
00:17:04,314 --> 00:17:05,115
TISSUE, DEMONSTRATED OR ASSUMED

442
00:17:05,115 --> 00:17:06,850
SOME COMING FROM ENTERIC NEURONS

443
00:17:06,850 --> 00:17:10,420
BUT TYLER WANTED TO ASK WHETHER

444
00:17:10,420 --> 00:17:12,789
THEY WERE CONTEXTS COMING FROM

445
00:17:12,789 --> 00:17:14,357
TUFT CELLS.

446
00:17:14,357 --> 00:17:17,794
HE ADOPTED THIS CLASSIC

447
00:17:17,794 --> 00:17:20,063
PHYSIOLOGY TOOL, USING CHAMBER,

448
00:17:20,063 --> 00:17:22,532
YOU HAVE TWO PHYSIOLOGICALLY

449
00:17:22,532 --> 00:17:24,935
BUFFERED CHAMBERS, INSERT ON

450
00:17:24,935 --> 00:17:25,802
THIS SLIDER HERE, INSERT

451
00:17:25,802 --> 00:17:29,072
EPITHELIUM OF CHOICE, IN THIS

452
00:17:29,072 --> 00:17:31,141
CASE SMALL INTESTINE HE PINNED

453
00:17:31,141 --> 00:17:32,976
UP ON, POLARIZED WITH THE VILLI

454
00:17:32,976 --> 00:17:35,412
ON ONE SIDE, BASAL SIDE ON THE

455
00:17:35,412 --> 00:17:35,745
OTHER.

456
00:17:35,745 --> 00:17:37,814
VOLTAGE CLAMP THIS AND MEASURE

457
00:17:37,814 --> 00:17:40,417
HOW MUCH CURRENT YOU NEED TO

458
00:17:40,417 --> 00:17:41,718
INJECT TO MAINTAIN THE SAME

459
00:17:41,718 --> 00:17:45,221
VOLTAGE, AND THAT CURRENT IS

460
00:17:45,221 --> 00:17:47,090
CALLED THE ISC, INDUCIBLE SHORT

461
00:17:47,090 --> 00:17:50,327
CIRCUIT, PROPORTIONAL TO ION

462
00:17:50,327 --> 00:17:51,394
MOVEMENT ACROSS EPITHELIUM.

463
00:17:51,394 --> 00:17:54,264
YOU'RE MEASURING IN REAL TIME

464
00:17:54,264 --> 00:17:57,801
ION FLUX ACROSS THE EPITHELIUM.

465
00:17:57,801 --> 00:18:01,137
THIS IS WHEN TYLER USES SMALL

466
00:18:01,137 --> 00:18:01,938
INTESTINE, BASELINE ISC,

467
00:18:01,938 --> 00:18:03,139
STIMULATES WITH SUCCINATE WHICH

468
00:18:03,139 --> 00:18:04,407
WE STARTED WITH BECAUSE IT WAS

469
00:18:04,407 --> 00:18:08,345
THE ONLY LIGAND WE HAD, YOU GET

470
00:18:08,345 --> 00:18:11,448
THIS VERY RAPID SPIKE IN THE

471
00:18:11,448 --> 00:18:14,384
ISC, HAPPENING, STARTING IN

472
00:18:14,384 --> 00:18:16,386
SECONDS, LASTS MINUTES, AND THEN

473
00:18:16,386 --> 00:18:18,121
DROPS BACK DOWN.

474
00:18:18,121 --> 00:18:19,489
IT IS KINETICALLY VERY SIMILAR

475
00:18:19,489 --> 00:18:24,127
TO WHAT HAPPENS IF YOU STIMULATE

476
00:18:24,127 --> 00:18:32,836
UP EPITHELIUM WITH ACETYLCHOLINE

477
00:18:32,836 --> 00:18:35,005
MIMIC CASH CHARBACHOL, COMPLEX

478
00:18:35,005 --> 00:18:35,305
WITH SODIUM.

479
00:18:35,305 --> 00:18:38,341
SOME EFFECT OF SODIUM ALONE.

480
00:18:38,341 --> 00:18:40,343
WE SWITCHED TO USING SYNTHETIC

481
00:18:40,343 --> 00:18:44,581
AGONIST OF THE SUCCINATE

482
00:18:44,581 --> 00:18:46,583
RECEPTOR THAT ELIMINATES THIS

483
00:18:46,583 --> 00:18:48,652
CHLORIDE COMPONENT OF THE

484
00:18:48,652 --> 00:18:49,185
RESPONSE.

485
00:18:49,185 --> 00:18:53,356
AND THEN THIS IS PUBLISHED, SO

486
00:18:53,356 --> 00:18:55,291
I'M GOING TO SUMMARIZE QUICKLY,

487
00:18:55,291 --> 00:19:01,264
HE WAS ABLE TO USE KNOCKOUTS TO

488
00:19:01,264 --> 00:19:04,934
SHOW THIS RESPONSE REQUIRED

489
00:19:04,934 --> 00:19:05,502
SUCCINATE RECEPTOR, REQUIRED

490
00:19:05,502 --> 00:19:07,303
TRPM 5 AGAIN, THAT KEY ION

491
00:19:07,303 --> 00:19:09,806
CHANNEL IN THE TUFT CELLS, AND

492
00:19:09,806 --> 00:19:12,742
AGAIN TUFT CELLS ARE THE ONLY

493
00:19:12,742 --> 00:19:17,013
EPITHELIAL CELLS THAT EXPRESS

494
00:19:17,013 --> 00:19:17,347
IT.

495
00:19:17,347 --> 00:19:21,818
THIS IS A KNOCKOUT WITHOUT TUFT

496
00:19:21,818 --> 00:19:23,520
CELLS, OTHERS ARE FINE.

497
00:19:23,520 --> 00:19:30,627
THE TISSUE WAS RESPONSING TO THE

498
00:19:30,627 --> 00:19:31,628
CHARBACHOL.

499
00:19:31,628 --> 00:19:32,395
THIS REQUIRES ACETYLCHOLINE FROM

500
00:19:32,395 --> 00:19:35,231
TUFT CELLS, USING A CRE WITH

501
00:19:35,231 --> 00:19:38,401
CHAT FLUX LINE, YOU LOSE

502
00:19:38,401 --> 00:19:42,338
RESPONSE TO THE LIGAND BUT NOT

503
00:19:42,338 --> 00:19:45,575
TO THE CHARBACHOL.

504
00:19:45,575 --> 00:19:51,414
SO, AGAIN, THERE'S TONS MORE

505
00:19:51,414 --> 00:19:53,650
CHARACTERIZATION TYLER IN.

506
00:19:53,650 --> 00:19:56,419
IN REAL CELL CONNECTED TUFT CELL

507
00:19:56,419 --> 00:19:57,320
FUNCTION, MANIPULATE WERE

508
00:19:57,320 --> 00:19:58,788
INHIBITORS AND KNOCKOUTS.

509
00:19:58,788 --> 00:20:02,325
HE WAS ABLE TO MAP THIS PATHWAY

510
00:20:02,325 --> 00:20:05,628
TO SHOW IT REQUIRED MUSCULAR

511
00:20:05,628 --> 00:20:12,435
ACETYLCHOLINE RECEPTORS,  TO SHW

512
00:20:12,435 --> 00:20:22,112
THIS WAS CHLORIDE AND SUSTAIN

513
00:20:22,112 --> 00:20:23,546
SECRETION YOU NEED UPTAKE, YOU

514
00:20:23,546 --> 00:20:25,181
NEED THE RESPONSE UNIT LINKING

515
00:20:25,181 --> 00:20:27,584
SENSING CAPACITY OF THE TUFT

516
00:20:27,584 --> 00:20:29,853
CELL TO INSTANTANEOUS SORT OF

517
00:20:29,853 --> 00:20:33,757
EFFECTOR FUNCTION OF FLUID

518
00:20:33,757 --> 00:20:35,759
SECRETION, FUNDAMENTAL ASPECT OF

519
00:20:35,759 --> 00:20:37,794
EPITHELIAL BIOLOGY AND

520
00:20:37,794 --> 00:20:39,329
PHYSIOLOGY, MUCOSAL BIOLOGY.

521
00:20:39,329 --> 00:20:40,830
AND IT'S REALLY CONSISTENT WITH

522
00:20:40,830 --> 00:20:42,932
EMERGING THEME THAT TUFT CELLS

523
00:20:42,932 --> 00:20:46,202
ARE USING ACETYLCHOLINE TO

524
00:20:46,202 --> 00:20:46,936
COMMUNICATE WITH NEIGHBORING

525
00:20:46,936 --> 00:20:49,506
EPITHELIAL CELLS, A BEAUTIFUL

526
00:20:49,506 --> 00:20:51,941
SERIES OF PAPERS THAT HAS

527
00:20:51,941 --> 00:20:54,544
SIMILARLY SHOWN THIS IN OTHER

528
00:20:54,544 --> 00:20:55,578
TISSUES LIKE GALLBLADDER AND

529
00:20:55,578 --> 00:20:55,812
AIRWAYS.

530
00:20:55,812 --> 00:20:59,349
BUT OF COURSE I TOLD YOU WE'RE

531
00:20:59,349 --> 00:21:00,884
TYPE 2 IMMUNITY LAB, WANTED TO

532
00:21:00,884 --> 00:21:02,285
KNOW IF THIS HAD ANYTHING TO DO

533
00:21:02,285 --> 00:21:04,621
WITH THE TYPE 2 IMMUNE RESPONSE.

534
00:21:04,621 --> 00:21:06,055
TO ADDRESS THAT LET ME EXPLAIN

535
00:21:06,055 --> 00:21:08,858
OUR CURRENT UNDERSTANDING OF HOW

536
00:21:08,858 --> 00:21:10,493
YOU ELIMINATE WORMS FROM

537
00:21:10,493 --> 00:21:10,794
INTESTINE.

538
00:21:10,794 --> 00:21:11,728
IT'S A TWO-STEP PROCESS WHERE

539
00:21:11,728 --> 00:21:14,063
THE FIRST IS A PRIMING PROCESS

540
00:21:14,063 --> 00:21:17,467
THAT TAKES IN THE CASE OF THIS

541
00:21:17,467 --> 00:21:19,068
HOOKWORM MODEL TAKES ABOUT SIX

542
00:21:19,068 --> 00:21:19,702
DAYS.

543
00:21:19,702 --> 00:21:21,371
AND THIS IS WHERE THE CIRCUIT IS

544
00:21:21,371 --> 00:21:21,871
SO IMPORTANT.

545
00:21:21,871 --> 00:21:24,541
YOU NEED TO SENSE THE WORMS,

546
00:21:24,541 --> 00:21:29,012
MAKE THESE EFFECTORS THAT

547
00:21:29,012 --> 00:21:29,813
ACTIVATE TYPE 2 SIGNALING,

548
00:21:29,813 --> 00:21:34,617
AMPLIFY THROUGH THE FEED FORWARD

549
00:21:34,617 --> 00:21:35,118
CIRCUIT.

550
00:21:35,118 --> 00:21:37,120
THERE'S ACUTE EVENT AROUND DAY

551
00:21:37,120 --> 00:21:39,589
7, CALLED WEEP AND SWEEP.

552
00:21:39,589 --> 00:21:45,862
THE WEEP IS INCREASED FLUID AND

553
00:21:45,862 --> 00:21:46,996
MUCUS PRODUCTION, DETACHES THE

554
00:21:46,996 --> 00:21:49,933
WORM, THE SWEEP IS MORE HYPER

555
00:21:49,933 --> 00:21:51,634
CONTRACTION OF THE SMOOTH MUSCLE

556
00:21:51,634 --> 00:21:53,670
TO EXPEL THEM OUT OF THE

557
00:21:53,670 --> 00:21:54,037
INTESTINE.

558
00:21:54,037 --> 00:21:55,438
WHAT'S NOTABLE THAT BOTH OF

559
00:21:55,438 --> 00:21:58,608
THESE PROCESSES CAN BE DIRECTLY

560
00:21:58,608 --> 00:22:00,743
DRIVEN BY ACETYLCHOLINE, AND

561
00:22:00,743 --> 00:22:03,246
WE'VE KNOWN FOR DECADES THAT

562
00:22:03,246 --> 00:22:07,784
WORMS SECRETE A LOT OF

563
00:22:07,784 --> 00:22:09,319
ACETYLCHOLINE ESTERASES, IF

564
00:22:09,319 --> 00:22:10,286
PATHOGEN IS ANTAGONIZING

565
00:22:10,286 --> 00:22:10,887
SOMETHING THAT'S PROBABLY

566
00:22:10,887 --> 00:22:17,794
IMPORTANT PART OF THE HOST

567
00:22:17,794 --> 00:22:18,094
DEFENSE.

568
00:22:18,094 --> 00:22:20,196
DO YOU NEED TUFT CELL

569
00:22:20,196 --> 00:22:21,798
ACETYLCHOLINE TO CLEAR WORMS OUT

570
00:22:21,798 --> 00:22:22,498
OF THE INTESTINE?

571
00:22:22,498 --> 00:22:27,136
YES, YOU DO.

572
00:22:27,136 --> 00:22:29,372
THIS IS DELETING ONLY FROM TUFT

573
00:22:29,372 --> 00:22:31,474
CELLS, ON DAY 5 THERE'S

574
00:22:31,474 --> 00:22:32,308
EQUIVALENT COLONIZATION, WHETHER

575
00:22:32,308 --> 00:22:34,344
OR NOT YOU HAVE CHAT.

576
00:22:34,344 --> 00:22:38,648
BUT BY DAY 7 AND 9, YOU SEE THAT

577
00:22:38,648 --> 00:22:39,949
THE WILD TYPE FUNCTIONAL MICE

578
00:22:39,949 --> 00:22:43,186
ARE CLEARING BUT CHAT DEFICIENT

579
00:22:43,186 --> 00:22:43,519
ARE DELAYED.

580
00:22:43,519 --> 00:22:43,920
OKAY.

581
00:22:43,920 --> 00:22:45,989
SO, WHAT IS IT THE TUFT CELLS

582
00:22:45,989 --> 00:22:46,522
ARE DOING?

583
00:22:46,522 --> 00:22:48,591
DO THEY HAVE ANY ROLE IN THAT

584
00:22:48,591 --> 00:22:52,095
PRIMING STEP IN THE CIRCUIT

585
00:22:52,095 --> 00:22:52,428
AMPLIFICATION?

586
00:22:52,428 --> 00:22:53,796
AND AGAIN TO SUMMARIZE QUICKLY

587
00:22:53,796 --> 00:22:55,231
THE ANSWER IS NO.

588
00:22:55,231 --> 00:22:56,666
BY ANY MEASURE TYLER COULD LOOK

589
00:22:56,666 --> 00:23:01,137
AT KIND OF THE IMMUNE

590
00:23:01,137 --> 00:23:02,038
ACTIVATION, WHETHER IT'S IL-13

591
00:23:02,038 --> 00:23:07,010
PRODUCTION, ILC 200, -- ILCs 2,

592
00:23:07,010 --> 00:23:12,649
EARLY OR LATER WHEN WE CAN'T

593
00:23:12,649 --> 00:23:14,350
PROCESS THE INTESTINE, DOESN'T

594
00:23:14,350 --> 00:23:16,619
MATTER FOR THE HYPERPLASIA OF

595
00:23:16,619 --> 00:23:20,723
TUFT CELLS, OF GOBLET CELLS,

596
00:23:20,723 --> 00:23:21,891
LENGTHENING OF INTESTINE SO WE

597
00:23:21,891 --> 00:23:23,960
DON'T THINK IT'S INVOLVED IN THE

598
00:23:23,960 --> 00:23:24,294
PRIMING STEP.

599
00:23:24,294 --> 00:23:25,929
THIS IS SUPPORTED BY EXPERIMENT

600
00:23:25,929 --> 00:23:27,964
WHERE TYLER DELAYED THE DELETION

601
00:23:27,964 --> 00:23:29,866
OF ACETYLCHOLINE PRODUCTION FROM

602
00:23:29,866 --> 00:23:31,734
TUFT CELLS UNTIL DAY 5, AND

603
00:23:31,734 --> 00:23:33,803
STILL FOUND DELAY IN CLEARANCE.

604
00:23:33,803 --> 00:23:36,306
THIS PLACES THE FUNCTION OF TUFT

605
00:23:36,306 --> 00:23:39,642
CELLS  ACETYLCHOLINE INTO THE

606
00:23:39,642 --> 00:23:40,944
LAST TWO DAYS OF THIS PARTICULAR

607
00:23:40,944 --> 00:23:41,444
WORM INFECTION.

608
00:23:41,444 --> 00:23:44,180
IS IT WEEP OR SWEEP?

609
00:23:44,180 --> 00:23:48,952
SAND FOR THIS WE WERE LUCKY TO

610
00:23:48,952 --> 00:23:50,320
COLLABORATE WITH TUAN, HERE IN

611
00:23:50,320 --> 00:23:51,955
THE AUDIENCE, ORGAN BATH SYSTEM

612
00:23:51,955 --> 00:23:55,792
SET UP, HE COULD MOUND -- FIRST

613
00:23:55,792 --> 00:23:58,995
EXPAND TUFT CELLS, INJECTING

614
00:23:58,995 --> 00:24:00,129
MICE WITH RECOMBINANT IL-25,

615
00:24:00,129 --> 00:24:01,297
MOUNT INTESTINE IN THE ORGAN

616
00:24:01,297 --> 00:24:05,134
BATH, TRY TO STIMULATE IT.

617
00:24:05,134 --> 00:24:09,305
IN THIS CASE DIRECTLY ACTIVATES

618
00:24:09,305 --> 00:24:10,006
THE TRPM5 CHANNEL.

619
00:24:10,006 --> 00:24:12,742
BUT WHEN THEY DID THAT THERE WAS

620
00:24:12,742 --> 00:24:16,279
NO CHANGE IN THE NUMBER OF PEAKS

621
00:24:16,279 --> 00:24:18,681
OR AMPLITUDE, NO CLEAR EVIDENCE

622
00:24:18,681 --> 00:24:21,517
ACTIVATING TOUGH CELLS ACUTELY

623
00:24:21,517 --> 00:24:23,820
ACTIVATED CONTRACTION.

624
00:24:23,820 --> 00:24:26,789
UNLIKE POSITIVE CONTROL

625
00:24:26,789 --> 00:24:27,323
ACETYLCHOLINE.

626
00:24:27,323 --> 00:24:29,392
TYLER DIDN'T SEE DIFFERENT IN

627
00:24:29,392 --> 00:24:30,159
TRANSIT TIME IN INFECTED MICE

628
00:24:30,159 --> 00:24:37,800
WHETHER OR NOT THERE WAS

629
00:24:37,800 --> 00:24:39,335
ACETYLCHOLINE.

630
00:24:39,335 --> 00:24:40,903
THIS COMPONENT, THE FLUID

631
00:24:40,903 --> 00:24:42,939
SECRETION, INCREASED AS MICE

632
00:24:42,939 --> 00:24:45,475
DEVELOPED HYPERPLASIA FROM DAY 0

633
00:24:45,475 --> 00:24:45,742
TO DAY 7.

634
00:24:45,742 --> 00:24:47,577
AND SO WHAT WE THINK IS THAT

635
00:24:47,577 --> 00:24:48,811
THIS MAY GIVE US SOME

636
00:24:48,811 --> 00:24:51,214
UNDERSTANDING OF AT LEAST SOME

637
00:24:51,214 --> 00:24:54,183
OF THE FUNCTION OF HYPERPLASIA.

638
00:24:54,183 --> 00:24:55,518
ACETYLCHOLINE DOES NOT SIGNAL

639
00:24:55,518 --> 00:24:57,854
FAR ONCE OUTSIDE OF THE CELL,

640
00:24:57,854 --> 00:25:07,363
THERE'S A LOT OF ESTERASES, IN

641
00:25:07,363 --> 00:25:08,431
HOMEOSTATIS, IT IS RARE,

642
00:25:08,431 --> 00:25:10,733
COVERAGE IS SPARSE.

643
00:25:10,733 --> 00:25:13,736
WHAT HYPERPLASIA DOES IS EXPANDS

644
00:25:13,736 --> 00:25:15,138
THE COVERAGE, REDUCES THE PLACES

645
00:25:15,138 --> 00:25:18,408
WHERE THE WORMS CAN HIDE, AND

646
00:25:18,408 --> 00:25:20,009
AMPLIFIES THIS WEEP RESPONSE

647
00:25:20,009 --> 00:25:21,344
WHICH THEN REACHES CRITICAL

648
00:25:21,344 --> 00:25:23,713
LEVEL AROUND AGAIN DAY 7 WHERE

649
00:25:23,713 --> 00:25:25,381
IT CAN CONTRIBUTE TO THE

650
00:25:25,381 --> 00:25:26,149
CLEARANCE.

651
00:25:26,149 --> 00:25:32,255
SO THAT'S HOW WE THINKING ABOUT

652
00:25:32,255 --> 00:25:34,590
SIGNALING TO NEIGHBORING CELLS.

653
00:25:34,590 --> 00:25:36,292
THIS IS THE GRAPHICAL SUMMARY

654
00:25:36,292 --> 00:25:43,666
FROM TYLER'S PAPER, THE  IDEA OF

655
00:25:43,666 --> 00:25:46,102
BASOLATERAL RELEASE, HE SHOWED

656
00:25:46,102 --> 00:25:51,874
IN THE USING CHAMBER DRIVING

657
00:25:51,874 --> 00:25:52,608
APICAL SECRETION, PUBLISHED WITH

658
00:25:52,608 --> 00:25:53,209
COMPLEMENTARY WORK.

659
00:25:53,209 --> 00:25:55,178
THEY DIDN'T FIND ANY ROLE IN

660
00:25:55,178 --> 00:25:57,046
THAT SORT OF INITIAL PRIMING.

661
00:25:57,046 --> 00:25:59,015
BUT THEY FOUND THERE'S ALSO

662
00:25:59,015 --> 00:26:00,850
APICAL SECRETION OF THE

663
00:26:00,850 --> 00:26:02,351
ACETYLCHOLINE, THAT CAN ACT

664
00:26:02,351 --> 00:26:05,021
DIRECTLY ON WORMS TO CAUSE SORT

665
00:26:05,021 --> 00:26:07,957
OF SPASTIC PARALYSIS THAT MAY

666
00:26:07,957 --> 00:26:13,663
FURTHER WEAKEN ABILITY TO

667
00:26:13,663 --> 00:26:15,164
COLONIZE INTESTINE.

668
00:26:15,164 --> 00:26:19,702
TWO WAYS TO AID IN EXPULSION OF

669
00:26:19,702 --> 00:26:21,104
WORMS.

670
00:26:21,104 --> 00:26:22,438
ALL RIGHT.

671
00:26:22,438 --> 00:26:25,808
SO, THAT WAS THE TUFT CELL AND

672
00:26:25,808 --> 00:26:27,009
EPITHELIAL PHYSIOLOGY STORY IS

673
00:26:27,009 --> 00:26:27,477
PUBLISHED.

674
00:26:27,477 --> 00:26:29,011
I'M GOING TO SWITCH GEARS TO

675
00:26:29,011 --> 00:26:30,613
TALK ABOUT UNPUBLISHED STORY

676
00:26:30,613 --> 00:26:35,718
WHICH WAS STARTED BY A WONDERFUL

677
00:26:35,718 --> 00:26:36,586
POSTDOC, ALICE STANBURY,

678
00:26:36,586 --> 00:26:38,187
CO-MENTORED BY MARION PEPPER AND

679
00:26:38,187 --> 00:26:40,490
LEFT A YEAR AGO, TYLER HELPED

680
00:26:40,490 --> 00:26:42,792
MOVE THE PROJECT FORWARD FOR

681
00:26:42,792 --> 00:26:43,159
TIME.

682
00:26:43,159 --> 00:26:45,862
BUT THIS CAME FROM WHEN ALOE

683
00:26:45,862 --> 00:26:47,697
JOINED THE LAB, EVERYTHING WE

684
00:26:47,697 --> 00:26:49,699
HAD DONE WAS ON SHORT TIME

685
00:26:49,699 --> 00:26:50,099
SCALES.

686
00:26:50,099 --> 00:26:51,767
REALLY ABOUT INNATE IMMUNITY,

687
00:26:51,767 --> 00:26:54,504
THINGS LIKE THE ION FLUX IN

688
00:26:54,504 --> 00:26:57,240
SECONDS, AND AT MOST A 7-DAY

689
00:26:57,240 --> 00:26:59,475
WORM INFECTION, DRIVEN MOSTLY BY

690
00:26:59,475 --> 00:27:00,776
INNATE LYMPHOID CELLS.

691
00:27:00,776 --> 00:27:03,679
AND SO TYLER -- ALOE WANTED TO

692
00:27:03,679 --> 00:27:06,315
THINK ABOUT, WELL, WHAT ABOUT IN

693
00:27:06,315 --> 00:27:07,183
CHRONIC INFECTION OR

694
00:27:07,183 --> 00:27:09,285
REINFECTION, WHICH REALLY IS THE

695
00:27:09,285 --> 00:27:13,623
MORE RELEVANT TO HUMAN BIOLOGY.

696
00:27:13,623 --> 00:27:14,690
WE'RE COMMONLY CHRONICALLY

697
00:27:14,690 --> 00:27:16,125
INFECTED WITH HELLMET, IN PARTS

698
00:27:16,125 --> 00:27:17,693
OF THE WORLD WHERE THAT'S

699
00:27:17,693 --> 00:27:19,762
COMMON, AND THERE ARE MANY

700
00:27:19,762 --> 00:27:20,563
DEWORMING PROGRAMS THAT

701
00:27:20,563 --> 00:27:22,465
ELIMINATE THEM TEMPORARILY BUT

702
00:27:22,465 --> 00:27:23,099
THERE'S REINFECTION.

703
00:27:23,099 --> 00:27:25,568
THOSE ARE THE KINDS OF THINGS

704
00:27:25,568 --> 00:27:29,305
THAT HAPPEN NATURALLY IN HUMANS.

705
00:27:29,305 --> 00:27:30,940
SOME QUESTIONS ALOE WANTED TO

706
00:27:30,940 --> 00:27:32,341
ADDRESS IS FIRST OF ALL NOW THAT

707
00:27:32,341 --> 00:27:37,113
WE HAVE A SENSE OF HYPERPLASIA

708
00:27:37,113 --> 00:27:38,414
THAT'S IMPORTANT, AS PART OF

709
00:27:38,414 --> 00:27:40,283
EFFECTOR FUNCTION HOW IS THAT

710
00:27:40,283 --> 00:27:42,385
REGULATED IN THESE CHRONIC AND

711
00:27:42,385 --> 00:27:43,386
SECONDARY INFECTIONS, ALSO WHAT

712
00:27:43,386 --> 00:27:45,821
IS THE RELATIVE CONTRIBUTION OF

713
00:27:45,821 --> 00:27:48,357
THESE INNATE LYMPHOID CELLS AND

714
00:27:48,357 --> 00:27:50,126
Th2 CELLS WHICH HAS BEEN A BIT

715
00:27:50,126 --> 00:27:51,260
OF UNRESOLVED QUESTION IN THE

716
00:27:51,260 --> 00:27:54,864
FIELD BECAUSE OF A LACK OF TOOLS

717
00:27:54,864 --> 00:27:56,832
TO REALLY SPECIFICALLY, WE HAVE

718
00:27:56,832 --> 00:28:00,002
A HARD TIME ELIMINATING ILC2s

719
00:28:00,002 --> 00:28:03,372
WITHOUT IMPACTING AND SEPARATING

720
00:28:03,372 --> 00:28:03,873
Th2s.

721
00:28:03,873 --> 00:28:06,442
THE OTHER THING IS WHETHER

722
00:28:06,442 --> 00:28:08,678
THERE'S ANY INTERACTION BETWEEN

723
00:28:08,678 --> 00:28:11,047
TUFT CELLS AND Th2s, AND

724
00:28:11,047 --> 00:28:13,816
SORT OF -- BECAUSE THE QUESTION

725
00:28:13,816 --> 00:28:14,951
WAS -- LET ME BACK UP.

726
00:28:14,951 --> 00:28:16,319
OUR UNDERSTANDING OF THE

727
00:28:16,319 --> 00:28:20,189
FUNCTION FOR TUFT CELLS IN THESE

728
00:28:20,189 --> 00:28:21,324
INNATE PATHWAYS IS THAT

729
00:28:21,324 --> 00:28:28,231
BASICALLY THEY ARE THE EYES OF

730
00:28:28,231 --> 00:28:29,198
ILC2s, THEY NEED THEM TO TELL

731
00:28:29,198 --> 00:28:31,067
THEM THE WORSE IS THERE.

732
00:28:31,067 --> 00:28:32,101
THEY HAVE AN ANTIGEN RECEPTOR,

733
00:28:32,101 --> 00:28:33,669
ONCE YOU HAVE THE SPECIFIC T

734
00:28:33,669 --> 00:28:36,572
CELLS IS THERE ANY NEED FOR TUFT

735
00:28:36,572 --> 00:28:37,006
CELL ANYMORE?

736
00:28:37,006 --> 00:28:38,140
ALL RIGHT.

737
00:28:38,140 --> 00:28:41,711
IN ORDER TO START TO THINK ABOUT

738
00:28:41,711 --> 00:28:49,652
THIS, ALOE TOOK ADVANTAGE OF

739
00:28:49,652 --> 00:28:53,856
THIS DIFFERENT WORM MODEL, I

740
00:28:53,856 --> 00:28:58,227
LEARNED RECENTLY IT'S ACTUALLY

741
00:28:58,227 --> 00:29:03,432
BACERII, VERY DIFFERENT SPECIES,

742
00:29:03,432 --> 00:29:08,337
POLY GYRUS IS IN WILD MICE BUT

743
00:29:08,337 --> 00:29:11,007
WE'RE WORKING WITH BACERRI.

744
00:29:11,007 --> 00:29:13,342
THIS IS A MOUSE-ADAPTED

745
00:29:13,342 --> 00:29:16,212
PARASITE, INAFFECTED THROUGH

746
00:29:16,212 --> 00:29:17,113
ORAL GAVAGE, ESTABLISHED CHRONIC

747
00:29:17,113 --> 00:29:21,484
INFECT OR CAN IF YOU DON'T CLEAR

748
00:29:21,484 --> 00:29:21,751
IT.

749
00:29:21,751 --> 00:29:26,656
LARVAE ENTER AND BURROW INTO THE

750
00:29:26,656 --> 00:29:28,257
SUBMUCOSA WHERE A GRANULOMA

751
00:29:28,257 --> 00:29:33,796
FORMS AROUND THEM, IMMUNE

752
00:29:33,796 --> 00:29:35,665
GRANULOMA, UNTIL 7 OR 8, BECOME

753
00:29:35,665 --> 00:29:38,968
ADULTS, CRAWL BACK OUT AND INTO

754
00:29:38,968 --> 00:29:41,804
THE LUMEN AND SPEND THEIR LIVES.

755
00:29:41,804 --> 00:29:42,838
IF YOU DON'T MANIPULATE, THEY

756
00:29:42,838 --> 00:29:45,775
ARE THERE FOR WEEKS OR MONTHS,

757
00:29:45,775 --> 00:29:47,743
IN B 6 MICE.

758
00:29:47,743 --> 00:29:54,016
BUT YOU CAN CLEAR THEM WITH

759
00:29:54,016 --> 00:29:55,351
BASIC ANTI-HELLMAN FIX, TWO

760
00:29:55,351 --> 00:29:56,419
WEEKS AFTER INFECTION, WAIT FOR

761
00:29:56,419 --> 00:30:00,056
THINGS TO SETTLE DOWN AND DO A

762
00:30:00,056 --> 00:30:04,593
CHALLENGE INFECTION, 28 DAYS

763
00:30:04,593 --> 00:30:06,062
AFTER CLEARANCE.

764
00:30:06,062 --> 00:30:08,364
THEY START INSIST THERE'S A

765
00:30:08,364 --> 00:30:09,565
BIGGER GRANULOMA, AND THE MODEL

766
00:30:09,565 --> 00:30:18,407
IN THE FIELD IS THIS IS THE SITE

767
00:30:18,407 --> 00:30:20,376
OF RESTRICTION, CAN TRIP AND

768
00:30:20,376 --> 00:30:26,716
KILL WORMS, LIMITED RE' MERGENS

769
00:30:26,716 --> 00:30:27,850
INTO THE LUMEN.

770
00:30:27,850 --> 00:30:32,188
WAIT 14 DAYS, DRUG CLEAR, WAIT

771
00:30:32,188 --> 00:30:33,389
28-35, CHALLENGE WITH SECONDARY,

772
00:30:33,389 --> 00:30:35,024
OFTEN LOOK AT WORMS IN THE

773
00:30:35,024 --> 00:30:38,828
INTESTINE 14 DAYS AFTER THAT.

774
00:30:38,828 --> 00:30:40,563
ONE OTHER IMPORTANT TOOL I WANT

775
00:30:40,563 --> 00:30:44,066
TO MENTION BEFORE SHOWING THE

776
00:30:44,066 --> 00:30:47,903
DATA ARE THESE LCR 1 KNOCKOUT

777
00:30:47,903 --> 00:30:49,105
MICE, IL-2 KNOCKOUTS, LUCKY TO

778
00:30:49,105 --> 00:30:56,212
GET THESE FROM JORGE AT PENN,

779
00:30:56,212 --> 00:30:59,615
THEY STARTED BY DOING WHAT THE

780
00:30:59,615 --> 00:31:01,784
HEADY LAB SPECIALIZES IN 3D

781
00:31:01,784 --> 00:31:03,886
GENOME MAPPING IN INNATE

782
00:31:03,886 --> 00:31:05,187
LYMPHOID CELLS, REALIZED THERE

783
00:31:05,187 --> 00:31:08,257
IS THIS DISTAL ENHANCER WHICH

784
00:31:08,257 --> 00:31:11,961
THEY CALLED LCR1 THAT IS OPEN

785
00:31:11,961 --> 00:31:15,131
AND USED ONLY IN ILC2s, OF THE

786
00:31:15,131 --> 00:31:17,800
ID2 GENE, IMPORTANT FOR A LOT OF

787
00:31:17,800 --> 00:31:19,502
LYMPHOCYTE DEVELOPMENT BUT THIS

788
00:31:19,502 --> 00:31:22,171
DISTAL ENHANCER IS ONLY ACTIVE

789
00:31:22,171 --> 00:31:24,306
IN ILC2s, SO WHEN THEY KNOCKED

790
00:31:24,306 --> 00:31:26,208
OUT THAT ENHANCER THEY FOUND

791
00:31:26,208 --> 00:31:29,011
THEY COULD STILL TURN NAIVE T

792
00:31:29,011 --> 00:31:31,080
CELLS INTO Th2 CELLS FINE, NO

793
00:31:31,080 --> 00:31:32,348
PROBLEMS, NO DEFECTS.

794
00:31:32,348 --> 00:31:34,817
BUT THERE'S A BIG DEFECT AND

795
00:31:34,817 --> 00:31:36,419
LOSS OF ILC2s ACROSS TISSUE,

796
00:31:36,419 --> 00:31:40,423
SO IN THE LUNG, FAT, SKIN,

797
00:31:40,423 --> 00:31:44,827
INTESTINE, HOMEOSTATICALLY, WAY,

798
00:31:44,827 --> 00:31:48,464
WAY FEWER ILC2s, A NUMBER

799
00:31:48,464 --> 00:31:51,367
FUNCTION AS KNOCKOUTS AND LET US

800
00:31:51,367 --> 00:31:56,639
DIE  -- DISSECT FUNCTION A LITTE

801
00:31:56,639 --> 00:31:56,806
BIT.

802
00:31:56,806 --> 00:31:59,775
ALOE STARTED BY MAPPING TUFT

803
00:31:59,775 --> 00:32:01,243
CELL HYPERPLASIA ACROSS TIME IN

804
00:32:01,243 --> 00:32:02,344
THIS MODEL.

805
00:32:02,344 --> 00:32:06,649
SO WE'RE MEASURING THE NUMBER OF

806
00:32:06,649 --> 00:32:09,785
TUFT CELL PER MILLIMETER CRYPT

807
00:32:09,785 --> 00:32:10,019
VILLUS.

808
00:32:10,019 --> 00:32:12,555
THIS IS THE BASELINE IN B 6

809
00:32:12,555 --> 00:32:12,755
MICE.

810
00:32:12,755 --> 00:32:14,590
INITIALLY NOTHING HAPPENS.

811
00:32:14,590 --> 00:32:17,493
THEN THERE IS A SMALL PEAK OF

812
00:32:17,493 --> 00:32:23,599
THE RESPONSE ON DAY 4, AND THEN

813
00:32:23,599 --> 00:32:26,302
FLATTENS OUT, ANOTHER PEAK DAY

814
00:32:26,302 --> 00:32:29,805
10, 12, 14, AND THEN IT FADES

815
00:32:29,805 --> 00:32:31,841
AGAIN LATER.

816
00:32:31,841 --> 00:32:34,643
THIS IS DIFFERENT, THERE'S A

817
00:32:34,643 --> 00:32:35,544
SINGLE PEAK BEFORE CLEARANCE, AS

818
00:32:35,544 --> 00:32:37,713
SOON AS THE WORMS ARE GONE IT

819
00:32:37,713 --> 00:32:38,280
DROPS.

820
00:32:38,280 --> 00:32:40,115
WE THINK THIS IS BASICALLY THE

821
00:32:40,115 --> 00:32:41,784
DAY 4 RESPONSE IS BASICALLY THE

822
00:32:41,784 --> 00:32:43,586
SENSING OF THE WORMS RIGHT WHEN

823
00:32:43,586 --> 00:32:44,553
THEY PENETRATE INITIALLY AND

824
00:32:44,553 --> 00:32:47,022
TAKES A FEW DAYS FOR THE TUFT

825
00:32:47,022 --> 00:32:48,791
CELL EXPANSION TO HAPPEN AND THE

826
00:32:48,791 --> 00:32:50,626
SECOND IS WHEN THEY REEMERGE

827
00:32:50,626 --> 00:32:53,195
INTO THE INTESTINE.

828
00:32:53,195 --> 00:32:54,964
AND THE RULES THAT GOVERN

829
00:32:54,964 --> 00:32:56,432
HYPERPLASIA SHIFT AS YOU MOVE

830
00:32:56,432 --> 00:32:57,766
THROUGH THIS INFECTION.

831
00:32:57,766 --> 00:33:00,336
SO IN THIS -- AT DAY 4 TIME

832
00:33:00,336 --> 00:33:04,206
POINT, EVERYTHING WORKS THE WAY

833
00:33:04,206 --> 00:33:10,312
WE ALREADY KNEW, YOU NEED

834
00:33:10,312 --> 00:33:15,885
ILC2s, PRCM AND TUFT 25,

835
00:33:15,885 --> 00:33:17,553
WITHOUT THAT NOTHING HAPPENS.

836
00:33:17,553 --> 00:33:22,525
YES, THERE IS STILL CONTRIBUTION

837
00:33:22,525 --> 00:33:24,994
FOR ILC2sA TRPM 5 BUT IT'S NOT

838
00:33:24,994 --> 00:33:25,461
COMPLETE.

839
00:33:25,461 --> 00:33:26,896
ALL OF THESE ARE BACK AT

840
00:33:26,896 --> 00:33:30,900
BASELINE, THERE'S NO RESPONSE AT

841
00:33:30,900 --> 00:33:31,100
ALL.

842
00:33:31,100 --> 00:33:33,369
NOW THERE ARE THESE WAYS FOR

843
00:33:33,369 --> 00:33:34,470
GETTING HYPERPLASIA THAT

844
00:33:34,470 --> 00:33:36,338
FUNCTION OUTSIDE OF THIS

845
00:33:36,338 --> 00:33:36,572
CIRCUIT.

846
00:33:36,572 --> 00:33:39,875
THE OTHER THING IS IF WE BLOCK

847
00:33:39,875 --> 00:33:41,777
RECRUITMENT OF NEW CELLS INTO

848
00:33:41,777 --> 00:33:45,848
THE TISSUE BY AT LEAST

849
00:33:45,848 --> 00:33:47,116
LYMPHOCYTES USING FGY 20 TO

850
00:33:47,116 --> 00:33:50,219
TRACK IN LYMPH NODES YOU ALSO

851
00:33:50,219 --> 00:33:51,654
BLUNT THIS TUFT CELL RESPONSE,

852
00:33:51,654 --> 00:33:54,023
STARTING TO SUGGEST IT'S NOW

853
00:33:54,023 --> 00:33:54,990
REGULATED BY RECRUITED CELLS

854
00:33:54,990 --> 00:33:58,827
LIKE T CELLS AND INDEED IF WE

855
00:33:58,827 --> 00:34:00,763
DEPLETE CD4 POSITIVE CELLS THEY

856
00:34:00,763 --> 00:34:03,132
ARE ABSOLUTELY REQUIRED NOW FOR

857
00:34:03,132 --> 00:34:03,699
THIS HYPERPLASIA.

858
00:34:03,699 --> 00:34:06,769
I DIDN'T SHOW BUT THEY ARE

859
00:34:06,769 --> 00:34:08,537
DISPENSABLE FOR THE INITIAL ONE.

860
00:34:08,537 --> 00:34:09,805
THIS IS ADAPTIVE IMMUNITY,

861
00:34:09,805 --> 00:34:10,272
RIGHT?

862
00:34:10,272 --> 00:34:14,043
YOU'RE SWITCHING FROM INNATE

863
00:34:14,043 --> 00:34:18,747
CELLS TO ADAPTIVE CELLS.

864
00:34:18,747 --> 00:34:22,718
IN SECONDARY INFECTION, CLEARED,

865
00:34:22,718 --> 00:34:23,919
WAITED, REINFECTED, SECOND

866
00:34:23,919 --> 00:34:25,721
INFECTION IS MORE RAPID

867
00:34:25,721 --> 00:34:26,488
HYPERPLASIA, GOES UP

868
00:34:26,488 --> 00:34:28,424
IMMEDIATELY, STAYS HIGH

869
00:34:28,424 --> 00:34:30,859
THROUGHOUT THE COURSE OF

870
00:34:30,859 --> 00:34:31,360
INFECTION.

871
00:34:31,360 --> 00:34:34,697
AND SO NOW WE CAN TRACK WHO IS

872
00:34:34,697 --> 00:34:36,098
MAKING IL-13 AT THESE TIME

873
00:34:36,098 --> 00:34:36,565
POINTS.

874
00:34:36,565 --> 00:34:38,601
WE START HERE AT DAY 2 AFTER

875
00:34:38,601 --> 00:34:41,103
PRIMARY INFECTION, THERE'S LOTS

876
00:34:41,103 --> 00:34:42,504
OF IL-13 POSITIVE CELLS, SMART

877
00:34:42,504 --> 00:34:47,376
13 REPORTER FROM THE LOCKSLY

878
00:34:47,376 --> 00:34:48,944
LAB, BUT ALMOST EXCLUSIVELY

879
00:34:48,944 --> 00:34:49,445
ILC2s.

880
00:34:49,445 --> 00:34:51,113
WE CLEAR, LET THE MICE SIT.

881
00:34:51,113 --> 00:34:53,115
EVERYTHING TURNS BACK OFF.

882
00:34:53,115 --> 00:34:54,683
NOBODY MAKING IL-13.

883
00:34:54,683 --> 00:34:56,619
SECONDARY INFECTION YOU GET NICE

884
00:34:56,619 --> 00:35:01,624
IL-13 RESPONSE BUT EVERYTHING IS

885
00:35:01,624 --> 00:35:03,659
FLIPPED, 90 OR MORE PERCENT OF

886
00:35:03,659 --> 00:35:05,794
THE CELLS ARE T CELLS, ABOUT 40

887
00:35:05,794 --> 00:35:10,132
TIMES MORE T CELLS MAKING 13

888
00:35:10,132 --> 00:35:14,470
THAN ILC2s, INNATE CELLS HAVE

889
00:35:14,470 --> 00:35:16,138
BEEN COMPLETELY SWAMPED.

890
00:35:16,138 --> 00:35:18,307
AND THE RULES AGAIN HAVE

891
00:35:18,307 --> 00:35:18,874
CHANGED.

892
00:35:18,874 --> 00:35:21,010
IF WE'RE NOW LOOKING AGAIN AT

893
00:35:21,010 --> 00:35:21,944
THE SECONDARY INFECTION, NOW

894
00:35:21,944 --> 00:35:25,114
THERE'S NO ROLE AT ALL WE CAN

895
00:35:25,114 --> 00:35:28,550
SEE FOR ILC2s, THE KNOCKOUTS,

896
00:35:28,550 --> 00:35:30,986
HYPERPLASIA IS NORMAL.

897
00:35:30,986 --> 00:35:34,156
STILL T CELL DEPEND, NO LONGER

898
00:35:34,156 --> 00:35:36,125
DEPEND ON CIRCULATING CELLS.

899
00:35:36,125 --> 00:35:37,860
IT HAS ESSENTIALLY REESTABLISHED

900
00:35:37,860 --> 00:35:41,330
A TISSUE INTRINSIC CIRCUIT BUT

901
00:35:41,330 --> 00:35:43,399
REPLACED INNATE ILC2s WITH

902
00:35:43,399 --> 00:35:44,967
RESIDENT Th2s.

903
00:35:44,967 --> 00:35:47,970
IN FACT, THIS RESIDENT CIRCUIT

904
00:35:47,970 --> 00:35:49,538
SEEMS TO BE SUFFICIENT TO CLEAR

905
00:35:49,538 --> 00:35:51,106
THE MICE.

906
00:35:51,106 --> 00:35:52,841
HERE WE'RE EITHER BLOCKING CD4

907
00:35:52,841 --> 00:35:58,881
FOR THE  DURATION OF SECONDARY R

908
00:35:58,881 --> 00:35:59,882
BLOCKING CELLS WITH 20.

909
00:35:59,882 --> 00:36:01,350
LET ME WALK YOU THROUGH THIS.

910
00:36:01,350 --> 00:36:05,821
THIS IS A PRIMARY INFECTION.

911
00:36:05,821 --> 00:36:07,790
AND THERE'S NO CLEARANCE OF

912
00:36:07,790 --> 00:36:08,123
WORMS.

913
00:36:08,123 --> 00:36:10,559
WE PUT IN 200, YOU ALWAYS GET

914
00:36:10,559 --> 00:36:12,428
OUT ABOUT 150.

915
00:36:12,428 --> 00:36:13,796
A SECONDARY INFECTION IN

916
00:36:13,796 --> 00:36:14,396
WILDTYPE UNMANIPULATED MOUSE,

917
00:36:14,396 --> 00:36:15,564
THIS IS THE CLEARANCE.

918
00:36:15,564 --> 00:36:18,534
THIS IS THE PROTECTIVE RESPONSE

919
00:36:18,534 --> 00:36:19,968
THAT DEVELOPS.

920
00:36:19,968 --> 00:36:21,236
TOTALLY CD4 DEPENDENT, KNOWN AND

921
00:36:21,236 --> 00:36:21,870
PUBLISHED.

922
00:36:21,870 --> 00:36:23,872
YOU STILL GET REALLY GOOD

923
00:36:23,872 --> 00:36:25,007
CLEARANCE WHEN YOU BLOCK

924
00:36:25,007 --> 00:36:26,875
RECRUITMENT OF OTHER CELLS.

925
00:36:26,875 --> 00:36:30,379
SO SUGGESTING THERE'S REALLY

926
00:36:30,379 --> 00:36:31,647
THIS TISSUE INTRINSIC MECHANISM.

927
00:36:31,647 --> 00:36:31,914
OKAY.

928
00:36:31,914 --> 00:36:33,182
COMING BACK TO THE FIRST

929
00:36:33,182 --> 00:36:35,417
QUESTION THEN HOW IS THE TUFT

930
00:36:35,417 --> 00:36:38,454
CELL HYPERPLASIA REGULATED, WHAT

931
00:36:38,454 --> 00:36:41,623
IS RELEVANT ROLE OF ILC2s AND

932
00:36:41,623 --> 00:36:44,259
Th2s, AS YOU MOVE THROUGH

933
00:36:44,259 --> 00:36:52,735
CHRONIC FRONT PRIMARY  INFECTION

934
00:36:52,735 --> 00:36:54,770
INTO SECONDARY THERE ARE

935
00:36:54,770 --> 00:36:56,605
DIFFERENT RULES, REPLACING THE

936
00:36:56,605 --> 00:36:58,707
CIRCUITS WITH Th2s, NOW IN

937
00:36:58,707 --> 00:37:02,277
THE TISSUE, CRITICAL FOR

938
00:37:02,277 --> 00:37:03,011
REGULATING GRANULOMAS, CAN ALSO

939
00:37:03,011 --> 00:37:07,015
REPLACE THE FUNCTION OF THE

940
00:37:07,015 --> 00:37:09,451
ILC2s FOR DRIVING TUFT CELL

941
00:37:09,451 --> 00:37:09,985
HYPERPLASIA.

942
00:37:09,985 --> 00:37:13,122
THIS LEADS US TO THE SECOND

943
00:37:13,122 --> 00:37:13,789
QUESTION.

944
00:37:13,789 --> 00:37:15,691
WHICH IS, NOW THAT YOU'VE PUT

945
00:37:15,691 --> 00:37:17,960
Th2s IN THE CELL, THEY HAVE

946
00:37:17,960 --> 00:37:19,528
AN ANTIGEN RECEPTOR, THERE'S

947
00:37:19,528 --> 00:37:21,396
LOTS OF ANTIGEN AVAILABLE FROM

948
00:37:21,396 --> 00:37:24,433
THESE WORMS AT LEAST INITIALLY

949
00:37:24,433 --> 00:37:26,435
IN THE TISSUE, IS THERE ANY ROLE

950
00:37:26,435 --> 00:37:29,371
ANYMORE FOR TUFT CELLS?

951
00:37:29,371 --> 00:37:30,739
THESE Th2 CELLS COULD FUNCTION

952
00:37:30,739 --> 00:37:34,710
WITHOUT ANY NEED FOR TUFT CELLS.

953
00:37:34,710 --> 00:37:36,812
BUT TURNS OUT THERE DEFINITELY

954
00:37:36,812 --> 00:37:37,579
IS.

955
00:37:37,579 --> 00:37:38,914
SO, THIS IS USING EITHER THE

956
00:37:38,914 --> 00:37:42,551
MICE THAT DON'T HAVE TUFT CELLS,

957
00:37:42,551 --> 00:37:46,321
THE KNOCKOUTS, OR THE MICE THAT

958
00:37:46,321 --> 00:37:50,626
LACK THE CHEMO SENSING CHANNEL

959
00:37:50,626 --> 00:37:52,494
OR DEPOLARIZING CHANNEL TRPM 5,

960
00:37:52,494 --> 00:37:53,228
PRIMARY FUNCTION DOESN'T MATTER

961
00:37:53,228 --> 00:37:55,130
WHETHER HAVE YOU TUFT CELLS, 14

962
00:37:55,130 --> 00:37:58,167
DAYS AFTER PRIMARY YOU HAVE 150,

963
00:37:58,167 --> 00:37:58,934
130 WORMS.

964
00:37:58,934 --> 00:38:00,903
WILDTYPE YOU CLEAR THAT.

965
00:38:00,903 --> 00:38:03,038
BUT MICE THAT DON'T HAVE

966
00:38:03,038 --> 00:38:04,206
SECONDARY INFECTION BUT DON'T

967
00:38:04,206 --> 00:38:06,341
HAVE TUFT CELLS OR TRPM 5

968
00:38:06,341 --> 00:38:08,310
THERE'S BASICALLY NO CLEARANCE

969
00:38:08,310 --> 00:38:11,747
AT ALL BY 14 DAYS AFTER

970
00:38:11,747 --> 00:38:13,782
SECONDARY INFECTION.

971
00:38:13,782 --> 00:38:15,851
AND THIS CLEARANCE ALSO DEPENDS

972
00:38:15,851 --> 00:38:18,954
ON THE KIND OF TUFT CELL

973
00:38:18,954 --> 00:38:24,827
EFFECTORS AND ARMAMENTS.

974
00:38:24,827 --> 00:38:33,769
IL-25 DEFICIENT MICE DON'T

975
00:38:33,769 --> 00:38:33,969
CLEAR.

976
00:38:33,969 --> 00:38:37,206
THE RECEPTOR HAS A ROLE, ILC2s

977
00:38:37,206 --> 00:38:41,710
HAVE A MINIMAL ROLE IN TERMS OF

978
00:38:41,710 --> 00:38:43,345
CLEARANCE OF SECONDARY

979
00:38:43,345 --> 00:38:43,612
INFECTION.

980
00:38:43,612 --> 00:38:44,646
ONE IMPORTANT CAVEAT WITH ALL OF

981
00:38:44,646 --> 00:38:47,049
THESE EXPERIMENTS IS THAT THESE

982
00:38:47,049 --> 00:38:49,184
ARE ALL MICE THAT ARE GLOBAL

983
00:38:49,184 --> 00:38:50,385
CONSTITUENT KNOCKOUTS THAT LACK

984
00:38:50,385 --> 00:38:52,487
THESE THINGS BOTH DURING PRIMARY

985
00:38:52,487 --> 00:38:55,924
INFECTION AND DURING SECONDARY.

986
00:38:55,924 --> 00:39:01,697
SO, ALOE CROSSED OUR IL-25 FLOX

987
00:39:01,697 --> 00:39:03,732
MICE TO INDUCIBLE VILLAIN CRE TO

988
00:39:03,732 --> 00:39:05,300
LEAVE THESE INTACT THROUGH

989
00:39:05,300 --> 00:39:09,805
PRIMARY AND MEMORY AND JUST

990
00:39:09,805 --> 00:39:11,773
BEFORE REINFECTION ADMINISTER TO

991
00:39:11,773 --> 00:39:13,375
TAMOXIFEN TO DELETE IL-25, ONLY

992
00:39:13,375 --> 00:39:15,077
EXPRESSED IN TUFT CELLS IN THE

993
00:39:15,077 --> 00:39:19,781
EPITHELIUM AND SHE COULD

994
00:39:19,781 --> 00:39:20,749
RECAPITULATE CLEARANCE DEFECT SO

995
00:39:20,749 --> 00:39:22,684
CLEARLY TUFT CELLS ARE DOING IN

996
00:39:22,684 --> 00:39:24,920
14 DAYS OF SECONDARY INFECTION

997
00:39:24,920 --> 00:39:28,690
THAT'S REALLY IMPORTANT FOR

998
00:39:28,690 --> 00:39:31,059
ELIMINATING -- I'M SO SORRY --

999
00:39:31,059 --> 00:39:33,829
ELIMINATING WORMS FROM THE

1000
00:39:33,829 --> 00:39:34,496
INTESTINE.

1001
00:39:34,496 --> 00:39:36,832
THE QUESTION IS WHAT IS IT TUFT

1002
00:39:36,832 --> 00:39:45,841
CELLS ARE DOING TO REGULATE THIS

1003
00:39:45,841 --> 00:39:47,442
BAKERI CLEARANCE IN SECONDARY

1004
00:39:47,442 --> 00:39:49,711
INFECTION. 

1005
00:39:49,711 --> 00:39:50,646
I TOLD BUT ACETYLCHOLINE

1006
00:39:50,646 --> 00:39:51,313
PRODUCTION, THAT COULD BE WHAT

1007
00:39:51,313 --> 00:39:53,148
THEY ARE DOING.

1008
00:39:53,148 --> 00:39:57,486
IT'S PROBABLY PART BUT NOT THE

1009
00:39:57,486 --> 00:40:06,762
ENTIRE  EXPLANATION, TYLER REPET

1010
00:40:06,762 --> 00:40:12,401
THE CELLS, THIS IS A REAL TREND

1011
00:40:12,401 --> 00:40:14,303
TOWARDS DECREASED CLEARANCE IN

1012
00:40:14,303 --> 00:40:15,170
THE DEFICIENT MICE.

1013
00:40:15,170 --> 00:40:17,606
ONE THING I WILL MENTION IS

1014
00:40:17,606 --> 00:40:20,509
WHENEVER WE GIVE MICE TAMOXIFEN

1015
00:40:20,509 --> 00:40:22,711
IT HAS SOME BROAD DAMPENING

1016
00:40:22,711 --> 00:40:24,579
EFFECT ON TYPE 2 IMMUNITY,

1017
00:40:24,579 --> 00:40:27,749
REALLY ANNOYING AND CONFOUNDING.

1018
00:40:27,749 --> 00:40:31,887
WE GIVE ALL OUR MICE TAMOXIFEN,

1019
00:40:31,887 --> 00:40:33,388
YOU'RE SEEING NOT GREAT

1020
00:40:33,388 --> 00:40:35,023
CLEARANCE HERE, WORSE CLEARANCE

1021
00:40:35,023 --> 00:40:37,359
WITHOUT THE CHAT.

1022
00:40:37,359 --> 00:40:39,494
AGAIN, THIS DOESN'T SEEM TO

1023
00:40:39,494 --> 00:40:40,729
EXPLAIN THE REALLY COMPLETE LACK

1024
00:40:40,729 --> 00:40:42,931
OF CLEARANCE WE SEE IN, SAY,

1025
00:40:42,931 --> 00:40:45,667
TUFT CELL KNOCKOUTS.

1026
00:40:45,667 --> 00:40:47,669
ALSO HAS NO EFFECT ON WORM

1027
00:40:47,669 --> 00:40:52,941
FITNESS IN TERMS OF FECUNDITY.

1028
00:40:52,941 --> 00:40:55,110
GIVEN THE CELLS ARE ESSENTIAL TO

1029
00:40:55,110 --> 00:40:59,014
THIS CLEARANCE OF THE SECONDARY

1030
00:40:59,014 --> 00:41:00,949
INFECTION ARE TUFT CELLS AND

1031
00:41:00,949 --> 00:41:03,418
Th2 CELLS TALKING TO EACH

1032
00:41:03,418 --> 00:41:03,685
OTHER?

1033
00:41:03,685 --> 00:41:05,187
AND IN ORDER TO THINK ABOUT THIS

1034
00:41:05,187 --> 00:41:12,761
I WANT TO TALK ABOUT HOW WE

1035
00:41:12,761 --> 00:41:14,429
THOUGHT ABOUT ILC2 ACTIVATION TO

1036
00:41:14,429 --> 00:41:16,631
TAKE OUR CUE FROM T CELLS WHERE

1037
00:41:16,631 --> 00:41:18,934
WE REALLY UNDERSTAND TCR

1038
00:41:18,934 --> 00:41:20,369
SIGNALING, TRANSCRIPTION FACTORS

1039
00:41:20,369 --> 00:41:22,804
IT ACTIVATES AND TARGET

1040
00:41:22,804 --> 00:41:24,206
CYTOKINES AND Th2 EFFECTOR

1041
00:41:24,206 --> 00:41:25,640
CELL, AND THERE'S BEEN BEAUTIFUL

1042
00:41:25,640 --> 00:41:33,081
WORK FROM JOHN O'SHEA AT THE NIH

1043
00:41:33,081 --> 00:41:37,786
FOR EXAMPLE SHOWING THEY ARE

1044
00:41:37,786 --> 00:41:39,021
TRANSCRIPTIONALLY ALMOST THE

1045
00:41:39,021 --> 00:41:40,589
SAME CELL.

1046
00:41:40,589 --> 00:41:42,491
THEY DON'T HAVE A T-CELL

1047
00:41:42,491 --> 00:41:44,393
RECEPTOR, HOW DO THEY ACHIEVE

1048
00:41:44,393 --> 00:41:45,160
THAT?

1049
00:41:45,160 --> 00:41:49,064
WE PROPOSE BY COMBINING IL-25

1050
00:41:49,064 --> 00:41:52,300
FOR NF-kappaB AND AP 1 AND

1051
00:41:52,300 --> 00:41:53,001
LEUKOTRIENES, SYNERGY IS POTENT,

1052
00:41:53,001 --> 00:41:54,436
AND TURNS OUT IN THE INTESTINE

1053
00:41:54,436 --> 00:41:57,205
THE TUFT CELLS CAN PROVIDE BOTH

1054
00:41:57,205 --> 00:41:58,140
OF THOSE.

1055
00:41:58,140 --> 00:42:00,008
NOW, WE'RE TAKING THAT IDEA AND

1056
00:42:00,008 --> 00:42:02,044
PUTTING IT BACK ON THE T CELL

1057
00:42:02,044 --> 00:42:02,711
AND SAYING, OKAY, WELL, COULD

1058
00:42:02,711 --> 00:42:05,013
YOU DO THE SAME THING IN THE T

1059
00:42:05,013 --> 00:42:08,016
CELL AND ELIMINATE -- REDUCE OR

1060
00:42:08,016 --> 00:42:10,452
BYPASS NEED FOR T-CELL RECEPTOR,

1061
00:42:10,452 --> 00:42:12,687
INNATE-LIKE FUNCTION OF AN

1062
00:42:12,687 --> 00:42:13,255
EFFECTOR T CELL.

1063
00:42:13,255 --> 00:42:16,858
AND THIS IS NOT A NEW IDEA.

1064
00:42:16,858 --> 00:42:18,593
WE WERE HEAVILY INSPIRED BY

1065
00:42:18,593 --> 00:42:20,128
FIRST I THINK FOUNDATIONAL WORK

1066
00:42:20,128 --> 00:42:25,200
DONE AGAIN HERE BY BILL PAUL

1067
00:42:25,200 --> 00:42:26,868
SHOWING THAT IL-33 AND OTHER

1068
00:42:26,868 --> 00:42:29,337
CYTOKINES IN THE FAMILY CAN

1069
00:42:29,337 --> 00:42:30,205
ACTIVATE EFFECTOR CELLS.

1070
00:42:30,205 --> 00:42:33,308
WORK DONE BY THE LOCKSLY LAB

1071
00:42:33,308 --> 00:42:34,843
SIMILARLY SHOWING THAT YOU HAVE

1072
00:42:34,843 --> 00:42:36,511
THIS SORT OF CHECKPOINT IN THE

1073
00:42:36,511 --> 00:42:40,749
TISSUE WHERE T CELLS ENTERING

1074
00:42:40,749 --> 00:42:43,452
GET ALARMIN SIGNALS LIKE IL-33

1075
00:42:43,452 --> 00:42:48,523
AND IN FAT AT PIERCE LAB, THEY

1076
00:42:48,523 --> 00:42:51,760
ARE REGULATED BY IL-33.

1077
00:42:51,760 --> 00:42:54,963
BUT WE HAVE FOUND GENERALLY ALSO

1078
00:42:54,963 --> 00:42:55,964
FOR ILC2s IT'S REALLY 25

1079
00:42:55,964 --> 00:42:57,165
THAT'S WAY MORE IMPORTANT THAN

1080
00:42:57,165 --> 00:42:58,800
THE SMALL INTESTINE SO THAT WAS

1081
00:42:58,800 --> 00:43:02,771
WHAT WE WANTED TO THINK ABOUT.

1082
00:43:02,771 --> 00:43:06,141
TAKING A STEP BACK FIRST THOUGH

1083
00:43:06,141 --> 00:43:07,809
WHAT ABOUT THE GENERATION AND

1084
00:43:07,809 --> 00:43:08,844
PRIMING OF THE T CELLS IN THE

1085
00:43:08,844 --> 00:43:09,544
FIRST PLACE?

1086
00:43:09,544 --> 00:43:13,782
IS THERE ANY ROLE FOR TUFT CELLS

1087
00:43:13,782 --> 00:43:15,050
OR FOR ILC2s, YOU KNOW, TO BE

1088
00:43:15,050 --> 00:43:16,651
HONEST I THOUGHT THERE WOULD BE

1089
00:43:16,651 --> 00:43:18,820
A ROLE BECAUSE WE KNOW IN MICE

1090
00:43:18,820 --> 00:43:20,589
THAT DON'T HAVE TUFT CELLS

1091
00:43:20,589 --> 00:43:22,624
TISSUE RESPONSE IS ALMOST

1092
00:43:22,624 --> 00:43:22,858
ABSENT.

1093
00:43:22,858 --> 00:43:24,826
BUT DESPITE THAT WE CANNOT FIND

1094
00:43:24,826 --> 00:43:27,062
ANY DEFECT IN T CELL PRIMING

1095
00:43:27,062 --> 00:43:28,930
LOOKING IN THE LYMPH NODE ON DAY

1096
00:43:28,930 --> 00:43:31,466
7 OF MICE THAT HAVE NO TUFT

1097
00:43:31,466 --> 00:43:34,503
CELLS, THE PERCENT OF Th2s,

1098
00:43:34,503 --> 00:43:36,571
GATA3 POSITIVE, IS THE SAME,

1099
00:43:36,571 --> 00:43:40,008
PRODUCTION OF IL-13 IS

1100
00:43:40,008 --> 00:43:40,675
UNCHANGED.

1101
00:43:40,675 --> 00:43:42,110
AND AGAIN ALSO ADDRESSING THIS

1102
00:43:42,110 --> 00:43:43,678
LONGSTANDING QUESTION ABOUT WHAT

1103
00:43:43,678 --> 00:43:49,084
IS THE ROLE OF ILC2s IN Th2

1104
00:43:49,084 --> 00:43:50,952
PRIMING, AT LEASE USING THIS

1105
00:43:50,952 --> 00:43:57,492
KNOCKOUT NO DEFECT, IN ANYTHING

1106
00:43:57,492 --> 00:43:57,826
THERE'S MORE.

1107
00:43:57,826 --> 00:43:58,426
CONSISTENT WITH THAT WORK WE

1108
00:43:58,426 --> 00:44:01,163
DON'T THINK THIS IS A STORY

1109
00:44:01,163 --> 00:44:02,964
ABOUT THE MAKING OF EFFECTOR T

1110
00:44:02,964 --> 00:44:04,833
CELLS, MIGHT HAVE A ROLE IN

1111
00:44:04,833 --> 00:44:05,133
MAINTENANCE.

1112
00:44:05,133 --> 00:44:07,002
WE HAVEN'T LOOKED AT THAT.

1113
00:44:07,002 --> 00:44:09,404
BUT WE FOCUSED ON THE ACTIVATION

1114
00:44:09,404 --> 00:44:10,539
OF THE EFFECTOR RESPONSE.

1115
00:44:10,539 --> 00:44:13,208
SO NOW IF YOU LOOK AT THOSE --

1116
00:44:13,208 --> 00:44:16,011
IF YOU MAKE MEMORY Th2 CELLS

1117
00:44:16,011 --> 00:44:19,614
OUT OF THESE MICE INFECTED,

1118
00:44:19,614 --> 00:44:20,749
CLEARED, RESTED, AND EVEN THOUGH

1119
00:44:20,749 --> 00:44:23,018
THIS IS A GUT RESTRICTED

1120
00:44:23,018 --> 00:44:24,519
INFECTION YOU FIND THESE CELLS

1121
00:44:24,519 --> 00:44:27,689
IN LOTS OF TISSUES, ANOTHER

1122
00:44:27,689 --> 00:44:28,089
INTERESTING TOPIC.

1123
00:44:28,089 --> 00:44:33,295
AND SIMILAR TO WHAT IS REPORTED

1124
00:44:33,295 --> 00:44:35,030
THERE'S HIGH EXPRESSION, IN THE

1125
00:44:35,030 --> 00:44:37,699
FAT, SOME IN THE LUNG.

1126
00:44:37,699 --> 00:44:41,203
NOT SO MUCH IN THE SMALL

1127
00:44:41,203 --> 00:44:44,105
INTESTINE, LAMINA PROPRIA HERE,

1128
00:44:44,105 --> 00:44:48,210
BUT INSTEAD THEY HAVE IL-25

1129
00:44:48,210 --> 00:44:49,511
RECEPTOR, IL7RB SPECIFICALLY,

1130
00:44:49,511 --> 00:44:53,815
SOME IN THE MESENTERIC LYMPH

1131
00:44:53,815 --> 00:44:55,217
NODES, BUT THE FAT WHICH IS

1132
00:44:55,217 --> 00:44:58,486
WRAPPED AROUND THE INTESTINE,

1133
00:44:58,486 --> 00:44:59,554
THOSE ARE DIFFERENT Th2s,

1134
00:44:59,554 --> 00:45:05,827
EXPRESS A LOT OF IL-33 RECEPTOR

1135
00:45:05,827 --> 00:45:07,195
BUT NOT 25 RECEPTOR.

1136
00:45:07,195 --> 00:45:09,030
WE DON'T HAVE AN ANTIBODY TO

1137
00:45:09,030 --> 00:45:13,602
LOOK AT PROTEIN BUT BY

1138
00:45:13,602 --> 00:45:15,604
SEQUENCING IT SEEMS THEY

1139
00:45:15,604 --> 00:45:18,139
UPREGULATE TO SOME EXTENT.

1140
00:45:18,139 --> 00:45:28,683
SO THE SMALL INTESTINAL PROPRIA

1141
00:45:29,851 --> 00:45:31,052
LAMINA, IF WE TAKE THESE CELLS

1142
00:45:31,052 --> 00:45:36,291
OUT AND PUT THEM IN A DISH AND

1143
00:45:36,291 --> 00:45:40,228
STIMULATE WE CAN DO THAT.

1144
00:45:40,228 --> 00:45:41,696
THIS IS POSITIVE CONTROL,

1145
00:45:41,696 --> 00:45:44,332
REACTIVATE 70% OF THE Th2s,

1146
00:45:44,332 --> 00:45:53,942
AND WHEN YOU COMBINE IL-25 AND

1147
00:45:53,942 --> 00:45:57,045
LEUKOTRINES, AND IN VIVO TOO IF

1148
00:45:57,045 --> 00:45:58,847
WE MAKE MEMORY CELLS THROUGH

1149
00:45:58,847 --> 00:46:01,549
THIS PROTOCOL AND THEN INSTEAD

1150
00:46:01,549 --> 00:46:02,250
OF REINFECTING -- WELL, WE

1151
00:46:02,250 --> 00:46:05,954
INFECT AS A POSITIVE CONTROL OR

1152
00:46:05,954 --> 00:46:10,258
JUST INFECT RECOMBINANT  RECOMB,

1153
00:46:10,258 --> 00:46:11,926
AFTER TWO DAYS MAKING THE AMOUNT

1154
00:46:11,926 --> 00:46:17,032
AFTER EVEN WITH JUST GIVING

1155
00:46:17,032 --> 00:46:18,066
RECOMBINANT CYTOKINE.

1156
00:46:18,066 --> 00:46:20,435
WHAT ABOUT IF WE ASK THE

1157
00:46:20,435 --> 00:46:21,469
OPPOSITE, SUFFICIENCY QUESTION,

1158
00:46:21,469 --> 00:46:24,472
IF WE STIMULATE THEM WITH

1159
00:46:24,472 --> 00:46:26,608
SIGNALS, WE CAN ACTIVE THEM, DO

1160
00:46:26,608 --> 00:46:29,711
YOU NEED TUFT CELLS AND SIGNALS

1161
00:46:29,711 --> 00:46:32,013
TO ACTIVATE Th2s, WE STARTED

1162
00:46:32,013 --> 00:46:34,182
AT EARLY TIME POINT AND EFFECT

1163
00:46:34,182 --> 00:46:36,184
HERE IS MODEST TO NONE.

1164
00:46:36,184 --> 00:46:38,486
SO WHEN THERE'S NO TUFT CELLS

1165
00:46:38,486 --> 00:46:40,922
THERE IS DEFINITELY DEFECT IN

1166
00:46:40,922 --> 00:46:42,123
IL-13 PRODUCTION BY Th2s BUT

1167
00:46:42,123 --> 00:46:45,126
DOESN'T SEEM TO RELY ON IL-25.

1168
00:46:45,126 --> 00:46:46,795
THIS IS VERY MYSTERIOUS.

1169
00:46:46,795 --> 00:46:50,532
WE DON'T REALLY KNOW YET WHAT IT

1170
00:46:50,532 --> 00:46:51,399
IS.

1171
00:46:51,399 --> 00:46:55,236
DOESN'T SEEM TO BE LEUKOTRIENES

1172
00:46:55,236 --> 00:46:56,905
EITHER, AND HYPERPLASIA IS FINE

1173
00:46:56,905 --> 00:46:58,039
WITHOUT IL-25 BUT YOU REMEMBER

1174
00:46:58,039 --> 00:46:59,240
HOW THIS WORM WORKS?

1175
00:46:59,240 --> 00:47:03,912
THIS IS WHEN THE WORM IS IN THE

1176
00:47:03,912 --> 00:47:05,046
TISSUE.

1177
00:47:05,046 --> 00:47:07,382
WE THINK TUFT CELL MONITOR THE

1178
00:47:07,382 --> 00:47:07,716
LUMEN.

1179
00:47:07,716 --> 00:47:13,588
T CELLS CAN GET TONS OF ANTIGEN

1180
00:47:13,588 --> 00:47:17,158
AND MAY NOT RELY ON TUFT CELLS.

1181
00:47:17,158 --> 00:47:18,626
WE WANTED TO REINVESTIGATE

1182
00:47:18,626 --> 00:47:24,165
GRANULOMA AND LUMINAL PHASE OF

1183
00:47:24,165 --> 00:47:26,167
WORM INFECTION SO THERE'S A LOT

1184
00:47:26,167 --> 00:47:26,968
HAPPENING HERE.

1185
00:47:26,968 --> 00:47:29,337
WHAT TYLER DID IN THIS

1186
00:47:29,337 --> 00:47:37,212
EXPERIMENT WAS COMPARED PRIMARY

1187
00:47:37,212 --> 00:47:39,614
GRANULOMA ON DAY 10 AFTER

1188
00:47:39,614 --> 00:47:40,081
PRIMARY AND SECONDARY.

1189
00:47:40,081 --> 00:47:41,649
THERE ARE EQUAL NUMBERS IN

1190
00:47:41,649 --> 00:47:42,584
EITHER CASE.

1191
00:47:42,584 --> 00:47:43,418
PRIMARY INFECTION THEY ARE ALL

1192
00:47:43,418 --> 00:47:44,786
IN LIKE THE FIRST FIVE

1193
00:47:44,786 --> 00:47:46,721
CENTIMETERS OF THE SMALL

1194
00:47:46,721 --> 00:47:48,289
INTESTINE, SECONDARY INFECTION

1195
00:47:48,289 --> 00:47:50,625
THEY SPREAD OUT MORE WHICH IS

1196
00:47:50,625 --> 00:47:51,393
INTERESTING, I HAVE NOTHING MORE

1197
00:47:51,393 --> 00:47:53,495
TO SAY ABOUT THAT THOUGH.

1198
00:47:53,495 --> 00:47:55,497
WE DON'T UNDERSTAND IT.

1199
00:47:55,497 --> 00:47:56,965
AND DEFINITELY THERE ARE WORMS

1200
00:47:56,965 --> 00:47:59,267
TRAPPED IN GRANULOMAS IN THE

1201
00:47:59,267 --> 00:48:00,935
SECONDSARY INFECTION BUT NOT ALL

1202
00:48:00,935 --> 00:48:01,770
OF THEM.

1203
00:48:01,770 --> 00:48:04,372
MAJORITY OF THE WORMS ARE

1204
00:48:04,372 --> 00:48:05,974
ACTUALLY BACK OUT IN THE LUMEN

1205
00:48:05,974 --> 00:48:07,942
AND ESCAPED OUT OF THE

1206
00:48:07,942 --> 00:48:09,110
GRANULOMA, NOT HAPPY WORMS.

1207
00:48:09,110 --> 00:48:10,779
THEY DON'T MAKE ANY EGGS.

1208
00:48:10,779 --> 00:48:15,083
BUT THEY DO GET BACK INTO THE

1209
00:48:15,083 --> 00:48:15,817
LUMEN.

1210
00:48:15,817 --> 00:48:17,252
AND THERE ARE ACTUALLY EQUAL

1211
00:48:17,252 --> 00:48:20,555
NUMBERS OF WORMS AT THIS DAY 10

1212
00:48:20,555 --> 00:48:22,957
TIME POINT ACROSS WILD TYPE AND

1213
00:48:22,957 --> 00:48:24,492
TUFT CELL KNOCKOUT MICE BUT FOUR

1214
00:48:24,492 --> 00:48:26,494
DAYS LATER THAT'S NOT THE CASE.

1215
00:48:26,494 --> 00:48:28,663
WILD TYPE MICE WILL HAVE

1216
00:48:28,663 --> 00:48:30,098
ELIMINATED WORMS, KNOCKOUTS NOT

1217
00:48:30,098 --> 00:48:32,100
AT ALL SUGGESTING THAT THERE'S

1218
00:48:32,100 --> 00:48:32,934
ANOTHER CLEARANCE EVENT THAT HAS

1219
00:48:32,934 --> 00:48:35,036
TO HAPPEN IN THE LAST DAYS AFTER

1220
00:48:35,036 --> 00:48:37,372
THE WORMS COME OUT OF THE

1221
00:48:37,372 --> 00:48:40,742
GRANULOMA AND THIS MIGHT BE WHEN

1222
00:48:40,742 --> 00:48:43,878
THIS TUFT CELL/T CELL CROSSTALK

1223
00:48:43,878 --> 00:48:45,013
COULD BE IMPORTANT.

1224
00:48:45,013 --> 00:48:47,782
INDEED WE FIND AGAIN A PARTIAL

1225
00:48:47,782 --> 00:48:51,052
BUT MORE CONVINCING ROLE FOR

1226
00:48:51,052 --> 00:48:53,054
TUFT CELLS IN REGULATING

1227
00:48:53,054 --> 00:48:54,556
Th2s AT LATE DAY 10 TIME

1228
00:48:54,556 --> 00:48:55,757
POINT AFTER SECONDARY INFECTION.

1229
00:48:55,757 --> 00:48:59,360
THIS IS FIRST LOOKING IN

1230
00:48:59,360 --> 00:49:09,904
MESENTERIC LYMPH NODE, SOMEWHAT

1231
00:49:11,539 --> 00:49:13,975
FEWER Th2s MAKE LESS I L.

1232
00:49:13,975 --> 00:49:16,044
WE WERE ABLE TO GET SECONDARY

1233
00:49:16,044 --> 00:49:16,311
INFECTION.

1234
00:49:16,311 --> 00:49:18,346
FREQUENCY OF THE CELLS IS NOT

1235
00:49:18,346 --> 00:49:19,747
THAT DIFFERENT BUT THE

1236
00:49:19,747 --> 00:49:20,982
PRODUCTION OF IL-13 IS DOWN WHEN

1237
00:49:20,982 --> 00:49:24,652
YOU DON'T HAVE TUFT CELLS OR

1238
00:49:24,652 --> 00:49:26,321
DON'T HAVE IL-25.

1239
00:49:26,321 --> 00:49:28,523
DEFINITELY PARTIAL PHENOTYPE BUT

1240
00:49:28,523 --> 00:49:28,723
REAL.

1241
00:49:28,723 --> 00:49:30,391
AND THEN LASTLY SIMILAR TO WHAT

1242
00:49:30,391 --> 00:49:33,695
WE DID WITH THE ACETYLCHOLINE WE

1243
00:49:33,695 --> 00:49:34,696
TIMED DELETION, LEFT EVERYTHING

1244
00:49:34,696 --> 00:49:37,365
INTACT UNTIL DAY 6 AFTER

1245
00:49:37,365 --> 00:49:39,934
SECONDARY INFECTION, AND YOU

1246
00:49:39,934 --> 00:49:41,135
AGAIN SEE DEFECT SUGGESTING --

1247
00:49:41,135 --> 00:49:42,303
SUPPORTING THE IDEA THERE'S A

1248
00:49:42,303 --> 00:49:44,839
ROLE LATE IN INFECTION FOR THE

1249
00:49:44,839 --> 00:49:47,442
TUFT CELLS.

1250
00:49:47,442 --> 00:49:47,675
OKAY.

1251
00:49:47,675 --> 00:49:50,745
SO SUMMARIZING THEN THIS PART,

1252
00:49:50,745 --> 00:49:56,084
TUFT CELLS AND ILC2s ARE 

1253
00:49:56,084 --> 00:49:58,853
DISPENSABLE FOR PRIMING IN THE

1254
00:49:58,853 --> 00:49:59,153
MODEL.

1255
00:49:59,153 --> 00:50:00,889
IN THE LUMEN THIS SEEMS TO

1256
00:50:00,889 --> 00:50:01,823
REQUIRE TUFT CELLS, WE'RE NOT

1257
00:50:01,823 --> 00:50:05,593
SAYING THERE'S NO ROLE FOR TCR

1258
00:50:05,593 --> 00:50:06,828
BUT THERE'S AN ADDITIVE

1259
00:50:06,828 --> 00:50:09,831
IMPORTANT FUNCTION FOR THESE

1260
00:50:09,831 --> 00:50:14,235
ALARMENS MADE BY THE TUFT CREST.

1261
00:50:14,235 --> 00:50:19,207
ESSENTIALLY IT'S A REWIRING OF

1262
00:50:19,207 --> 00:50:21,709
CIRCUIT, TUFT Th2 CIRCUIT AND

1263
00:50:21,709 --> 00:50:25,079
WE THINK THIS HAS INTERESTING

1264
00:50:25,079 --> 00:50:26,848
IMPLICATIONS, YOU NEED TO

1265
00:50:26,848 --> 00:50:27,615
ESTABLISH TISSUE RESIDENT

1266
00:50:27,615 --> 00:50:31,119
RESPONSES IF YOU WANT A GOOD

1267
00:50:31,119 --> 00:50:37,292
VACCINE, POSSIBLY THROUGH A

1268
00:50:37,292 --> 00:50:39,994
PRIME POLL APPROACH.

1269
00:50:39,994 --> 00:50:41,796
IF THERE'S MISMATCHED ANTIGEN

1270
00:50:41,796 --> 00:50:44,499
TUFT CELLS SEEM TO SENSE

1271
00:50:44,499 --> 00:50:47,635
HELMINTHS, MAYBE THEY COULD DO

1272
00:50:47,635 --> 00:50:48,303
ACTIVATING OF BYSTANDER

1273
00:50:48,303 --> 00:50:50,772
Th2s, DOES THAT HAVE

1274
00:50:50,772 --> 00:50:52,173
CONSEQUENCE IN FOOD ALLERGY?

1275
00:50:52,173 --> 00:50:55,043
LAST THING I WANT TO SAY IS WE

1276
00:50:55,043 --> 00:50:58,413
THINK THIS IS NOT THE WHOLE

1277
00:50:58,413 --> 00:51:03,351
STORY SO WE'VE IDENTIFIED

1278
00:51:03,351 --> 00:51:04,385
ACETYLCHOLINE, EPITHELIUM,

1279
00:51:04,385 --> 00:51:09,324
TALKING TO ILC2s, IL-25

1280
00:51:09,324 --> 00:51:14,329
TALKING, COULD BE A RULE FOR

1281
00:51:14,329 --> 00:51:15,997
LEUKOTRIENES, WE HAVE REASONS TO

1282
00:51:15,997 --> 00:51:17,565
THINK IN SECONDARY INFECTION

1283
00:51:17,565 --> 00:51:19,133
MODEL THERE ARE OTHER TARGETS OF

1284
00:51:19,133 --> 00:51:20,234
TUFT CELL EFFECTORS.

1285
00:51:20,234 --> 00:51:21,502
THERE'S LOTS OF CELLS TO THINK

1286
00:51:21,502 --> 00:51:22,870
ABOUT HERE.

1287
00:51:22,870 --> 00:51:25,440
NEURONS, MYELOID CELLS, B CELLS,

1288
00:51:25,440 --> 00:51:28,042
THOSE ARE THINGS WE'RE JUST

1289
00:51:28,042 --> 00:51:29,744
STARTING TO DO.

1290
00:51:29,744 --> 00:51:30,144
OKAY.

1291
00:51:30,144 --> 00:51:31,579
SO BIG THANK YOU TO THE LAB.

1292
00:51:31,579 --> 00:51:34,515
IT'S BEEN WONDERFUL TO WORK WITH

1293
00:51:34,515 --> 00:51:35,083
THESE PEOPLE.

1294
00:51:35,083 --> 00:51:36,784
MOST OF THE PEOPLE WHO DID THE

1295
00:51:36,784 --> 00:51:38,386
WORK I SHOWED YOU TODAY HAVE

1296
00:51:38,386 --> 00:51:39,721
MOVED ON.

1297
00:51:39,721 --> 00:51:41,322
WE'VE GOT NEW GRADUATE STUDENTS,

1298
00:51:41,322 --> 00:51:42,824
I'M REALLY EXCITED TO SEE WHAT

1299
00:51:42,824 --> 00:51:45,126
THEY START UP.

1300
00:51:45,126 --> 00:51:46,527
LOTS OF FANTASTIC COLLABORATORS.

1301
00:51:46,527 --> 00:51:47,895
RIGHT, I WILL SAY WE'VE DONE

1302
00:51:47,895 --> 00:51:50,031
REALLY WITH FUNDING THIS YEAR,

1303
00:51:50,031 --> 00:51:52,367
WE HAVE A FUNDED POSTDOC

1304
00:51:52,367 --> 00:51:54,535
POSITION, $79,000 IN SEATTLE

1305
00:51:54,535 --> 00:51:56,504
NOW, I THINK THAT MIGHT BE THE

1306
00:51:56,504 --> 00:51:58,806
HIGHEST IN THE COUNTRY, A REALLY

1307
00:51:58,806 --> 00:52:00,541
COOL PROGRAM THAT WAS STARTED IN

1308
00:52:00,541 --> 00:52:02,010
THE DEPARTMENT THAT PROVIDES --

1309
00:52:02,010 --> 00:52:05,580
YOU CAN CHOOSE ACADEMIC OR

1310
00:52:05,580 --> 00:52:08,416
INDUSTRY TRACK FOR TRAINING AS A

1311
00:52:08,416 --> 00:52:08,650
POSTDOC.

1312
00:52:08,650 --> 00:52:11,619
SO LET ME KNOW IF YOU'RE

1313
00:52:11,619 --> 00:52:12,954
INTERESTED OR KNOW ANYONE, SEND

1314
00:52:12,954 --> 00:52:13,655
ME AN E-MAIL.

1315
00:52:13,655 --> 00:52:14,689
THANKS FOR YOUR ATTENTION.

1316
00:52:14,689 --> 00:52:17,558
HAPPY TO TAKE QUESTIONS. 

1317
00:52:17,558 --> 00:52:21,829
[APPLAUSE]

1318
00:52:21,829 --> 00:52:26,801
 

1319
00:52:26,801 --> 00:52:28,903
>> THAT WAS WONDERFUL, AS
USUAL.

1320
00:52:28,903 --> 00:52:30,538
TWO QUESTIONS.

1321
00:52:30,538 --> 00:52:35,843
THE FIRST QUESTION WAS TO DO

1322
00:52:35,843 --> 00:52:38,446
WITH ACETYLCHOLINE WEEP OR SWEEP

1323
00:52:38,446 --> 00:52:39,113
RESPONSE.

1324
00:52:39,113 --> 00:52:41,082
WONDERING IF IT APPLIES TO THE

1325
00:52:41,082 --> 00:52:45,987
LARGE 

1326
00:52:45,987 --> 00:52:48,189
LARGE AND SMALL INTESTINE, IF

1327
00:52:48,189 --> 00:52:50,091
YOU TRIED TRICURIOUS INFECTION

1328
00:52:50,091 --> 00:52:50,725
WHICH AFFECTS LARGE INTESTINE,

1329
00:52:50,725 --> 00:52:54,896
WHETHER OR NOT YOU GET THIS PART

1330
00:52:54,896 --> 00:52:56,531
OF THE EFFECTOR PATHWAY

1331
00:52:56,531 --> 00:52:57,365
ACTIVATION?

1332
00:52:57,365 --> 00:52:59,267
>> YEAH. 

1333
00:52:59,267 --> 00:53:01,135
WE'VE DONE PRELIMINARY

1334
00:53:01,135 --> 00:53:05,506
EXPERIMENTS WITH MY LAB MATE AND

1335
00:53:05,506 --> 00:53:07,742
NEIGHBOR, AND WORKS ON THIS.

1336
00:53:07,742 --> 00:53:09,143
IT'S BEEN CONFUSING, WE'VE DONE

1337
00:53:09,143 --> 00:53:13,815
IT THREE TIMES IN TUFT CELL

1338
00:53:13,815 --> 00:53:18,953
KNOCKOUTS TWICE, ONCE REQUIRED,

1339
00:53:18,953 --> 00:53:21,689
ONCE NOT.

1340
00:53:21,689 --> 00:53:24,058
IF YOU STIMULATE YOU GET FLUID

1341
00:53:24,058 --> 00:53:26,060
TISSUE RESPONSE TYLER SHOWED IN

1342
00:53:26,060 --> 00:53:27,228
COLONIC TISSUE.

1343
00:53:27,228 --> 00:53:29,063
BUT WHAT DOES NOT HAPPEN IS

1344
00:53:29,063 --> 00:53:30,998
COLON, YOU DO NOT GET

1345
00:53:30,998 --> 00:53:33,367
HYPERPLASIA, TO THE SPEND IT

1346
00:53:33,367 --> 00:53:35,369
CAN'T BE AMPLIFIED THE SAME WAY.

1347
00:53:35,369 --> 00:53:38,806
SO WE'RE NOT QUITE SURE HOW TO

1348
00:53:38,806 --> 00:53:40,408
THINK ABOUT THAT.

1349
00:53:40,408 --> 00:53:40,942
>> OKAY.

1350
00:53:40,942 --> 00:53:43,411
SO THE OTHER QUESTION I HAD WAS

1351
00:53:43,411 --> 00:53:46,581
TO DO WITH SECONDARY INFECTION.

1352
00:53:46,581 --> 00:53:51,152
AND, YOU KNOW, THE DATA YOU SHOW

1353
00:53:51,152 --> 00:53:53,354
ON THE REQUIREMENT FOR CD4 CELLS

1354
00:53:53,354 --> 00:53:57,525
AND INNATE CELLS REALLY LOOKS

1355
00:53:57,525 --> 00:53:59,260
LIKE IL-4 ACTIVATED MACROPHAGES,

1356
00:53:59,260 --> 00:54:06,400
IN THE SENSE THAT, YOU KNOW,

1357
00:54:06,400 --> 00:54:09,804
LIKE EARLY ON IN WOUND HEALING

1358
00:54:09,804 --> 00:54:13,508
YOU HAVE THIS T CELL INDEPENDENT

1359
00:54:13,508 --> 00:54:15,777
ACTIVATION OF MACROPHAGES BUT

1360
00:54:15,777 --> 00:54:19,814
LIKE SUSTAINED CHRONIC INFECTION

1361
00:54:19,814 --> 00:54:21,582
REQUIRES CD4 POSITIVE, MY

1362
00:54:21,582 --> 00:54:24,652
QUESTION HAVE YOU LOOKED AT

1363
00:54:24,652 --> 00:54:25,920
MAYBE MACROPHAGES BEING LIKE

1364
00:54:25,920 --> 00:54:28,923
THIS KIND OF PART OF THIS

1365
00:54:28,923 --> 00:54:31,292
CIRCUIT BETWEEN THE TUFT CELLS

1366
00:54:31,292 --> 00:54:33,394
AND THE Th2 CELLS AND, YOU

1367
00:54:33,394 --> 00:54:39,767
KNOW, JUST IN TERMS OF LIKE

1368
00:54:39,767 --> 00:54:42,870
MAYBE NUMBERS OF THESE ACTIVATED

1369
00:54:42,870 --> 00:54:44,238
MACROPHAGES BECAUSE THEY ARE

1370
00:54:44,238 --> 00:54:46,374
IMPORTANT IN CLEARANCE.

1371
00:54:46,374 --> 00:54:46,808
>> TOTALLY.

1372
00:54:46,808 --> 00:54:47,742
YEAH, BASICALLY THE ANSWER IS

1373
00:54:47,742 --> 00:54:49,811
NO, WE HAVEN'T DONE MUCH.

1374
00:54:49,811 --> 00:54:53,080
ONE THING WE DID WAS USE LYSE M

1375
00:54:53,080 --> 00:54:55,049
CRE TO TAKE THE 25 RECEPTOR OFF

1376
00:54:55,049 --> 00:54:57,318
MYELOID CELLS, JUST AS ONE --

1377
00:54:57,318 --> 00:55:00,755
>> AT THAT POINT I THINK
IT'S

1378
00:55:00,755 --> 00:55:02,089
THE -- WHAT'S ACTIVATING THOSE

1379
00:55:02,089 --> 00:55:04,826
CELLS IS ACTUALLY THE TH 2

1380
00:55:04,826 --> 00:55:05,092
CELLS.

1381
00:55:05,092 --> 00:55:05,993
>> I AGREE.

1382
00:55:05,993 --> 00:55:07,328
WE DIDN'T SEE ANYTHING WHEN WE

1383
00:55:07,328 --> 00:55:09,130
DID THAT.

1384
00:55:09,130 --> 00:55:09,397
>> YEAH.

1385
00:55:09,397 --> 00:55:12,433
>> THE OTHER THINGLY ADMIT
WE'VE

1386
00:55:12,433 --> 00:55:13,868
HAD TOTAL IL-13 BLINDERS, WE

1387
00:55:13,868 --> 00:55:14,836
LOVE TO USE REPORTERS, THAT

1388
00:55:14,836 --> 00:55:16,671
MEANS WE'VE GOT TO CROSS THEM

1389
00:55:16,671 --> 00:55:18,973
INTO ALL THESE THINGS.

1390
00:55:18,973 --> 00:55:20,541
WE'VE DONE EVERYTHING WITH 13

1391
00:55:20,541 --> 00:55:20,975
REPORTERS.

1392
00:55:20,975 --> 00:55:24,378
WE'RE NOW GEARING UP TO LOOK AND

1393
00:55:24,378 --> 00:55:25,179
SEE, IT WOULD BE REALLY

1394
00:55:25,179 --> 00:55:27,615
INTERESTING TO ME IF THERE'S A

1395
00:55:27,615 --> 00:55:29,750
BIGGER DEPENDENCE ON THE TUFT

1396
00:55:29,750 --> 00:55:30,952
CELLS FOR THE IL-4 PRODUCTION

1397
00:55:30,952 --> 00:55:32,520
THAN 13 PRODUCTION BY THE

1398
00:55:32,520 --> 00:55:36,891
Th2s WHICH THEY DO UNIQUELY,

1399
00:55:36,891 --> 00:55:39,493
ILC2s DON'T DO THAT.

1400
00:55:39,493 --> 00:55:45,199
IT MIGHT BE TRUE BECAUSE AGAIN

1401
00:55:45,199 --> 00:55:51,505
THIS LIKE M FAT SIGNAL FROM THE

1402
00:55:51,505 --> 00:55:51,973
LEUKOTRIENES.

1403
00:55:51,973 --> 00:55:52,173
TBD.

1404
00:55:52,173 --> 00:55:55,109
DO WE GO OVER HERE?

1405
00:55:55,109 --> 00:55:57,278
>>  A QUESTION ABOUT THE

1406
00:55:57,278 --> 00:55:59,013
ACETYLCHOLINE EFFECT, IN SOME

1407
00:55:59,013 --> 00:56:00,681
CASES YOU HAVE EFFECTS IN TUFT

1408
00:56:00,681 --> 00:56:03,618
CELLS, OTHER ONES YOU DON'T.

1409
00:56:03,618 --> 00:56:06,621
BUT ARE THERE OTHER SOURCES OF

1410
00:56:06,621 --> 00:56:09,390
ACETYLCHOLINE AND WHAT ABOUT

1411
00:56:09,390 --> 00:56:19,834
NERVOUS TISSUE, CAN YOU DO

1412
00:56:24,972 --> 00:56:25,172
VAGOTOMY.

1413
00:56:25,172 --> 00:56:27,508
>> WE DID A CRUDE ABLATION
OF

1414
00:56:27,508 --> 00:56:30,578
SENSORY NEURONS, THAT DID NOT

1415
00:56:30,578 --> 00:56:32,680
ALTER CLEARANCE.

1416
00:56:32,680 --> 00:56:35,049
>> IS THERE ROLE FOR

1417
00:56:35,049 --> 00:56:35,716
ACETYLCHOLINE IN DRIVING

1418
00:56:35,716 --> 00:56:40,922
DIFFERENT T-CELL RESPONSE, EARLY

1419
00:56:40,922 --> 00:56:42,423
OR LATE?

1420
00:56:42,423 --> 00:56:44,592
MACROPHAGES DO EXPRESS --

1421
00:56:44,592 --> 00:56:45,192
>> THIS NON-NEURONAL

1422
00:56:45,192 --> 00:56:50,331
ACETYLCHOLINE IS LIKE A HUGE NEW

1423
00:56:50,331 --> 00:56:52,099
FIELD, T CELLS, MACROPHAGES.

1424
00:56:52,099 --> 00:56:55,903
WE'VE NOT SEEN A ROLE FOR TUFT

1425
00:56:55,903 --> 00:56:56,871
CELL ACETYLCHOLINE BUT NEURONAL

1426
00:56:56,871 --> 00:56:57,638
CERTAINLY THAT'S POSSIBLE.

1427
00:56:57,638 --> 00:56:59,407
>> CAN YOU JUST BLOCK?

1428
00:56:59,407 --> 00:57:05,079
WHAT IF YOU BLOCK DURING A LATE

1429
00:57:05,079 --> 00:57:07,081
PHASE, FOR EXAMPLE?

1430
00:57:07,081 --> 00:57:12,620
>>  WITH LIKE AN ATROPINE?

1431
00:57:12,620 --> 00:57:13,454
>> WHY.

1432
00:57:13,454 --> 00:57:18,893
>> THE MICE ARE NOT HAPPY.

1433
00:57:18,893 --> 00:57:21,095
>> THANK YOU.

1434
00:57:21,095 --> 00:57:21,996
>> G-PROTEIN COUPLED
RECEPTOR

1435
00:57:21,996 --> 00:57:24,765
PATHWAY IS THAT INVOLVED IN

1436
00:57:24,765 --> 00:57:25,800
SENSING THE WORMS?

1437
00:57:25,800 --> 00:57:32,773
>>  WELL, SUCCINATE
RECEPTOR

1438
00:57:32,773 --> 00:57:34,508
PATHWAY IS NOT.

1439
00:57:34,508 --> 00:57:41,549
LIKE A G NET 3 KNOCKOUT MOUSE,

1440
00:57:41,549 --> 00:57:44,151
THAT --

1441
00:57:44,151 --> 00:57:44,952
>> GUSTISIN.

1442
00:57:44,952 --> 00:57:46,620
>> YES, NO SENSING,
TOTALLY

1443
00:57:46,620 --> 00:57:47,888
NORMAL CLEANS IN SENSING OF THE

1444
00:57:47,888 --> 00:57:48,356
WORMS.

1445
00:57:48,356 --> 00:57:51,559
>> THAT MOUSE DID HAVE A
GUT

1446
00:57:51,559 --> 00:57:54,628
PHENOTYPE IF I REMEMBER RIGHTLY,

1447
00:57:54,628 --> 00:58:00,267
HYPERINFLAMMATORY IN DSS MODEL.

1448
00:58:00,267 --> 00:58:02,236
DIDN'T MAKE IL-13 OR 5, THE

1449
00:58:02,236 --> 00:58:05,406
KNOCKOUT, YEAH. 

1450
00:58:05,406 --> 00:58:07,375
I COULD BE MISREMEMBERING BUT

1451
00:58:07,375 --> 00:58:09,977
I'M PRETTY SURE IT DID HAVE GUT

1452
00:58:09,977 --> 00:58:10,478
DEFECT.

1453
00:58:10,478 --> 00:58:12,747
>> I THINK THERE ARE
MULTIPLE G

1454
00:58:12,747 --> 00:58:14,482
NET KNOCKOUTS, ONE LINE HAD

1455
00:58:14,482 --> 00:58:15,416
SOMETHING, THE OTHER DIDN'T, IF

1456
00:58:15,416 --> 00:58:15,850
I REMEMBER.

1457
00:58:15,850 --> 00:58:17,284
I DON'T KNOW.

1458
00:58:17,284 --> 00:58:20,688
THIS IS LIKE TALKING TO PEOPLE

1459
00:58:20,688 --> 00:58:22,890
AT LIKE MONELLE WHO WORKED ON

1460
00:58:22,890 --> 00:58:23,624
THESE THINGS.

1461
00:58:23,624 --> 00:58:24,358
MEMORY, I THINK THEY DECIDED

1462
00:58:24,358 --> 00:58:27,795
THAT WAS NOT.

1463
00:58:27,795 --> 00:58:30,197
>> IS THERE MANY ORPHAN
GPCRs,

1464
00:58:30,197 --> 00:58:32,967
THIS WAS OUR APPROACH TO LOOK

1465
00:58:32,967 --> 00:58:36,070
JUST ON TUFT CELLS AND NOT

1466
00:58:36,070 --> 00:58:36,637
OTHERS.

1467
00:58:36,637 --> 00:58:37,738
FIRST NON-ORPHAN, LIKE TASTE

1468
00:58:37,738 --> 00:58:41,509
RECEPTORS WE DON'T SEE ON

1469
00:58:41,509 --> 00:58:42,576
INTESTINAL TUFT CELLS, THEY ARE

1470
00:58:42,576 --> 00:58:45,413
IN THE AIRWAY FOR SURE.

1471
00:58:45,413 --> 00:58:48,816
THERE'S ONE OR TWO, MOST

1472
00:58:48,816 --> 00:58:52,253
INTERESTING GPRC 5C, MADE A

1473
00:58:52,253 --> 00:58:53,888
KNOCKOUT, NO PHENOTYPE.

1474
00:58:53,888 --> 00:58:57,792
YEAH, WE'RE STILL SEARCHING.

1475
00:58:57,792 --> 00:59:00,795
>> THANKS.

1476
00:59:00,795 --> 00:59:01,796
 

1477
00:59:01,796 --> 00:59:03,531
>> GREAT TALK.

1478
00:59:03,531 --> 00:59:05,800
THINKING ABOUT AMPLIFICATION

1479
00:59:05,800 --> 00:59:07,902
CIRCUIT, ILCs OR Th2s, I

1480
00:59:07,902 --> 00:59:11,072
LIKE THE IDEA OF THE

1481
00:59:11,072 --> 00:59:12,139
ACETYLCHOLINE AS LIMITED

1482
00:59:12,139 --> 00:59:14,375
DIFFUSION RATE, DO YOU HAVE ANY

1483
00:59:14,375 --> 00:59:16,777
SENSE IS THERE A DISADVANTAGE TO

1484
00:59:16,777 --> 00:59:18,679
HAVING PARASITE INDUCED LEVELS

1485
00:59:18,679 --> 00:59:21,182
OF TUFT CELLS AT HOMEOSTASIS IN

1486
00:59:21,182 --> 00:59:22,383
THE ABSENCE OF TYPE 2 INITIATOR

1487
00:59:22,383 --> 00:59:23,951
MEANING IF YOU HAVE TOO MANY

1488
00:59:23,951 --> 00:59:30,024
AROUND DOES THAT INTERFERE WITH

1489
00:59:30,024 --> 00:59:30,825
NORMAL ABSORPTIVE FUNCTION?

1490
00:59:30,825 --> 00:59:31,292
>>  ZERO CONSEQUENCE.

1491
00:59:31,292 --> 00:59:33,060
THIS IS WORK THAT I WAS INVOLVED

1492
00:59:33,060 --> 00:59:36,464
IN THE PROJECT, LIKE BEFORE I

1493
00:59:36,464 --> 00:59:37,531
LEFT THE LOCKSLY LAB, CHRIS

1494
00:59:37,531 --> 00:59:39,366
SNYDER FINISHED IT.

1495
00:59:39,366 --> 00:59:43,704
WE MADE THESE MICE, DELETED A

1496
00:59:43,704 --> 00:59:46,340
20, THIS BROAD INHIBITOR, OUT OF

1497
00:59:46,340 --> 00:59:48,075
ILC2s, IT TOOK THE BRAKES OFF

1498
00:59:48,075 --> 00:59:49,276
THE CIRCUIT.

1499
00:59:49,276 --> 00:59:54,882
THOSE MICE AS SOON AS PROTEUS

1500
00:59:54,882 --> 00:59:56,784
EXPANDED THEY HAVE MASSIVE

1501
00:59:56,784 --> 00:59:59,386
HIGHER PLASIA, ZERO HEALTH

1502
00:59:59,386 --> 01:00:00,221
CONSEQUENCES, NO WEIGHT

1503
01:00:00,221 --> 01:00:03,858
SEQUENCES, PUT THEM IN METABOLIC

1504
01:00:03,858 --> 01:00:07,895
CAGES, NO DIFFERENCES, DID

1505
01:00:07,895 --> 01:00:10,030
CALORIEMETRY TO ABSORB THE SAME.

1506
01:00:10,030 --> 01:00:13,701
INTESTINE WAS 27% LONGER, THE

1507
01:00:13,701 --> 01:00:14,368
ONLY DIFFERENCE.

1508
01:00:14,368 --> 01:00:16,036
THE IDEA WAS THAT BECAUSE ON

1509
01:00:16,036 --> 01:00:26,547
LIKE A PER AREA BASIS THEY HAD

1510
01:00:28,782 --> 01:00:31,085
FEWER ABSORBTIVE DEVELOPS,

1511
01:00:31,085 --> 01:00:32,086
WITHIN DAYS OF ACTIVATING TYPE 2

1512
01:00:32,086 --> 01:00:35,222
IMMUNITY IN THE SMALL INTESTINE

1513
01:00:35,222 --> 01:00:38,225
IT GETS 20% LONGER, REALLY COOL,

1514
01:00:38,225 --> 01:00:39,693
THE MECHANISMS OF THAT ARE NOT

1515
01:00:39,693 --> 01:00:42,763
CLEAR TO ME, YEAH.

1516
01:00:42,763 --> 01:00:45,833
 

1517
01:00:45,833 --> 01:00:46,700
 

1518
01:00:46,700 --> 01:00:48,135
>> HI.

1519
01:00:48,135 --> 01:00:50,804
SO, I WAS WONDERING ABOUT

1520
01:00:50,804 --> 01:00:55,809
THIS -- IN THE SECOND PRIME

1521
01:00:55,809 --> 01:00:57,378
MODEL DESCRIBED LIKE INITIAL --

1522
01:00:57,378 --> 01:00:59,346
SUSTAINED MORE TUFT CELLS, YOU

1523
01:00:59,346 --> 01:01:02,049
SEE MORE TUFT CELLS, DO YOU

1524
01:01:02,049 --> 01:01:03,317
THINK SUSTAINED RESPONSES, THE

1525
01:01:03,317 --> 01:01:05,152
SAME TUFT CELLS THAT ARE

1526
01:01:05,152 --> 01:01:07,321
DIVIDING OR IS IT STEM CELLS?

1527
01:01:07,321 --> 01:01:09,790
BECAUSE YOU KIND OF -- WHEN YOU

1528
01:01:09,790 --> 01:01:11,325
INTRODUCED THIS IDEA YOU

1529
01:01:11,325 --> 01:01:12,826
MENTIONED THIS HAPPENS IN FOUR

1530
01:01:12,826 --> 01:01:15,129
DAYS AND GOES BACK TO NORMAL, DO

1531
01:01:15,129 --> 01:01:16,697
YOU THINK IT'S THE SAME TUFT

1532
01:01:16,697 --> 01:01:19,200
CELLS OR JUST STEM CELL AND TO

1533
01:01:19,200 --> 01:01:23,070
BE HONEST I DON'T KNOW HOW YOU

1534
01:01:23,070 --> 01:01:24,305
DISSECT THAT.

1535
01:01:24,305 --> 01:01:27,374
>> YEAH, EVERYTHING WE'VE
DONE

1536
01:01:27,374 --> 01:01:30,344
TO LOOK AT TUFT CELL LIFESPAN

1537
01:01:30,344 --> 01:01:31,378
SUGGESTS THEY DON'T -- ACTUALLY

1538
01:01:31,378 --> 01:01:35,082
JUST IN THE LAST MONTH WE FOUND

1539
01:01:35,082 --> 01:01:37,284
EVIDENCE THEY LIVE A LITTLE

1540
01:01:37,284 --> 01:01:38,352
LONGER THAN ENTEROCYTES BUT ONLY

1541
01:01:38,352 --> 01:01:39,553
A FEW DAYS, IF YOU DON'T KEEP

1542
01:01:39,553 --> 01:01:41,055
MAKING NEW ONES THEY ARE GOING

1543
01:01:41,055 --> 01:01:43,190
TO BE GONE PRETTY QUICKLY.

1544
01:01:43,190 --> 01:01:48,596
SO THERE MUST BE A CONTINUAL

1545
01:01:48,596 --> 01:01:49,797
ADDITIONAL PRODUCTION, YEAH,

1546
01:01:49,797 --> 01:01:50,097
YEAH.

1547
01:01:50,097 --> 01:01:51,966
>> THANK YOU.

1548
01:01:51,966 --> 01:01:52,433
>> HI.

1549
01:01:52,433 --> 01:01:53,267
GREAT TALK.

1550
01:01:53,267 --> 01:02:00,507
I WAS WONDERING IF YOU SEE ANY

1551
01:02:00,507 --> 01:02:06,814
ROLE FOR -- IN YOUR RESPONSE OF

1552
01:02:06,814 --> 01:02:08,649
THE RESPONSE -- I DON'T KNOW HOW

1553
01:02:08,649 --> 01:02:11,085
MANY BASOPHILS THERE ARE.

1554
01:02:11,085 --> 01:02:13,554
DO THEY ACCUMULATE DURING THE

1555
01:02:13,554 --> 01:02:13,821
INFECTION?

1556
01:02:13,821 --> 01:02:20,094
>> YEAH, WE'VE NOT DONE
ANY

1557
01:02:20,094 --> 01:02:21,061
CHARACTERIZATION OF BASOPHILS IN

1558
01:02:21,061 --> 01:02:21,762
THAT MODEL.

1559
01:02:21,762 --> 01:02:23,130
OTHERS HAVE.

1560
01:02:23,130 --> 01:02:26,867
THE CONCLUSION, THERE'S A ROLE

1561
01:02:26,867 --> 01:02:32,339
FOR BASOPHILS IN THE TRICURUS

1562
01:02:32,339 --> 01:02:34,074
MODEL, IN THE OTHER MODEL NOBODY

1563
01:02:34,074 --> 01:02:39,546
FOUND A ROLE FOR BASOPHILS TO MY

1564
01:02:39,546 --> 01:02:40,214
KNOWLEDGE.

1565
01:02:40,214 --> 01:02:40,681
>> WONDERFUL, JAKOB.

1566
01:02:40,681 --> 01:02:43,584
I LIKE TO ASK BIG QUESTIONS.

1567
01:02:43,584 --> 01:02:48,188
>> I THINK TO THINK ABOUT
THEM.

1568
01:02:48,188 --> 01:02:50,624
>> DID GOD MAKE INTESTINAL
TUFT

1569
01:02:50,624 --> 01:02:54,528
CELLS TO EXPEL HELMINTHS OR IS

1570
01:02:54,528 --> 01:02:56,196
THERE ANOTHER FUNCTION, I WOULD

1571
01:02:56,196 --> 01:02:59,700
PUT MONEY ON ANOTHER HOMEOSTATIC

1572
01:02:59,700 --> 01:02:59,967
FUNCTION.

1573
01:02:59,967 --> 01:03:03,203
SECOND PART, ANN WAS DELVES IN

1574
01:03:03,203 --> 01:03:04,838
THE SAME DIRECTION, IS WHAT ARE

1575
01:03:04,838 --> 01:03:07,541
THEY REALLY SENSING?

1576
01:03:07,541 --> 01:03:11,512
IF IT'S NOT SUCCINATE OR IF IT

1577
01:03:11,512 --> 01:03:13,814
IS HELMINTHS THE ONLY THING THEY

1578
01:03:13,814 --> 01:03:15,149
REALLY SEE WHAT COULD IT

1579
01:03:15,149 --> 01:03:17,885
POSSIBLY BE THEY ARE

1580
01:03:17,885 --> 01:03:18,185
RECOGNIZING?

1581
01:03:18,185 --> 01:03:19,753
>>  OKAY, YOUR BIG
QUESTION I

1582
01:03:19,753 --> 01:03:20,387
REALLY LIKE THAT ONE.

1583
01:03:20,387 --> 01:03:22,256
I'VE THOUGHT ABOUT IT.

1584
01:03:22,256 --> 01:03:23,590
WE'VE THOUGHT ABOUT IT.

1585
01:03:23,590 --> 01:03:24,925
TOTALLY AGREE WITH YOU.

1586
01:03:24,925 --> 01:03:26,894
I DON'T THINK THESE CELLS

1587
01:03:26,894 --> 01:03:29,596
EVOLVED FIRST TO SENSE WORMS.

1588
01:03:29,596 --> 01:03:31,565
AND MY MOTIVATION FOR THAT IS

1589
01:03:31,565 --> 01:03:33,000
GOBLET CELLS.

1590
01:03:33,000 --> 01:03:35,703
GOBLET CELLS ARE CRITICAL FOR

1591
01:03:35,703 --> 01:03:39,807
THE DEFENSE AGAINST HELMINTHS

1592
01:03:39,807 --> 01:03:40,974
BUT I DON'T THINK ANYONE WOULD

1593
01:03:40,974 --> 01:03:43,210
ARGUE THEY EVOLVED FOR THAT.

1594
01:03:43,210 --> 01:03:48,515
YOU NEED TO MAKE MUCUS FOR

1595
01:03:48,515 --> 01:03:49,817
EPITHELIAL MUCOSAL, AND THEY

1596
01:03:49,817 --> 01:03:50,184
WERE RECRUITED.

1597
01:03:50,184 --> 01:03:53,587
MY BIAS IS TO THINK IT'S SIMILAR

1598
01:03:53,587 --> 01:03:56,523
FOR TUFT CELLS, AND IT'S WHY I

1599
01:03:56,523 --> 01:03:57,891
LOVE THE PHYSIOLOGY AND FLUID

1600
01:03:57,891 --> 01:04:04,565
STORY BECAUSE IT DUST LINK TO

1601
01:04:04,565 --> 01:04:05,799
FUNDAMENTAL PHYSIOLOGY OF

1602
01:04:05,799 --> 01:04:06,266
MUCOSAL BARRIERS.

1603
01:04:06,266 --> 01:04:07,935
IN TUFT CELL KNOCKOUTS THERE'S

1604
01:04:07,935 --> 01:04:12,373
NO BIG -- IT'S NOT LIKE THEIR

1605
01:04:12,373 --> 01:04:13,874
INTESTINE TRIES OUT.

1606
01:04:13,874 --> 01:04:14,975
THESE ARE PATHWAYS THAT ARE

1607
01:04:14,975 --> 01:04:16,143
IMPORTANT, A LOT OF BACKUP,

1608
01:04:16,143 --> 01:04:17,845
MAYBE THERE'S SOMETHING LIKE

1609
01:04:17,845 --> 01:04:19,780
THAT.

1610
01:04:19,780 --> 01:04:21,515
BUT I AGREE, I MEAN I THINK,

1611
01:04:21,515 --> 01:04:22,783
YEAH, I AGREE I DON'T THINK

1612
01:04:22,783 --> 01:04:24,318
THAT'S WHAT THEY EVOLVED FOR

1613
01:04:24,318 --> 01:04:25,319
FIRST AND THEY WERE RECRUITED

1614
01:04:25,319 --> 01:04:26,987
INTO THAT BECAUSE THEY TURNED

1615
01:04:26,987 --> 01:04:30,824
OUT TO BE USEFUL. 

1616
01:04:30,824 --> 01:04:31,558
WHAT ARE THEY SENSING?

1617
01:04:31,558 --> 01:04:37,164
>>  THEY PROBABLY WOULDN'T
SEE

1618
01:04:37,164 --> 01:04:37,698
PARASITES ANYWAY?

1619
01:04:37,698 --> 01:04:38,766
>>  TISSUES?

1620
01:04:38,766 --> 01:04:40,634
EFFECTORS FUNCTIONS SEEM TO BE

1621
01:04:40,634 --> 01:04:46,507
CONSERVED BUT SENSING IS 

1622
01:04:46,507 --> 01:04:48,642
DIVERSIFIED, WE DON'T THINK THEY

1623
01:04:48,642 --> 01:04:52,479
SENSE WORMS IN THE AIRWAYS,

1624
01:04:52,479 --> 01:04:53,781
SENSING BACTERIA OR IRRITANTS.

1625
01:04:53,781 --> 01:04:58,252
WHAT ARE THEY SENSING FROM

1626
01:04:58,252 --> 01:04:58,752
WORMS?

1627
01:04:58,752 --> 01:05:00,187
YEAH, THIS HAS BEEN THE

1628
01:05:00,187 --> 01:05:00,454
CHALLENGE.

1629
01:05:00,454 --> 01:05:01,822
WE STILL DON'T KNOW.

1630
01:05:01,822 --> 01:05:03,857
WORMS SECRETE A LOT OF STUFF, A

1631
01:05:03,857 --> 01:05:08,429
LOT OF CANDIDATES, BUT YEAH.

1632
01:05:08,429 --> 01:05:09,797
>> REALLY INTERESTING

1633
01:05:09,797 --> 01:05:10,130
PRESENTATION.

1634
01:05:10,130 --> 01:05:14,768
THE DID MY Ph.D. WITH CHRIS

1635
01:05:14,768 --> 01:05:19,173
ALLEN AT UCSF, I LIKE TO ASK

1636
01:05:19,173 --> 01:05:20,541
SMALL QUESTIONS.

1637
01:05:20,541 --> 01:05:22,609
JUST WANTED TO GIVE YOU SOME

1638
01:05:22,609 --> 01:05:33,120
SPACE TO MAYBE EXPLAIN WHY YOU

1639
01:05:35,756 --> 01:05:35,989
ALLERGIES.

1640
01:05:35,989 --> 01:05:39,927
WHEN YOU MENTIONED FLUID LOSS

1641
01:05:39,927 --> 01:05:41,028
FROM THE CHLORIDE IONS, CHOLERA

1642
01:05:41,028 --> 01:05:43,497
TOXIN WHICH WE USE TO INDUCE

1643
01:05:43,497 --> 01:05:44,465
FOOD ALLERGY IN MICE, WHETHER

1644
01:05:44,465 --> 01:05:47,668
YOU MIGHT HAVE LOOKED AT CHOLERA

1645
01:05:47,668 --> 01:05:50,304
TOXIN INFECTION AND DO TUFT

1646
01:05:50,304 --> 01:05:51,505
CELLS EXPAND AFTER THAT?

1647
01:05:51,505 --> 01:05:54,208
>>  THAT WAS A LOT OF
SMALL

1648
01:05:54,208 --> 01:05:55,175
QUESTIONS.

1649
01:05:55,175 --> 01:05:55,476
[LAUGHTER]

1650
01:05:55,476 --> 01:05:56,543
SUPER INTERESTING.

1651
01:05:56,543 --> 01:05:58,412
YEAH, YOU IDENTIFIED A BIT OF

1652
01:05:58,412 --> 01:06:00,280
NARRATIVE FLAW IN MY

1653
01:06:00,280 --> 01:06:01,482
PRESENTATION THAT I HIGHLIGHT

1654
01:06:01,482 --> 01:06:02,649
ALLERGENS AND DON'T TALK ABOUT

1655
01:06:02,649 --> 01:06:02,916
THEM.

1656
01:06:02,916 --> 01:06:06,053
I DON'T THINK TUFT CELLS SENSE

1657
01:06:06,053 --> 01:06:06,487
ALLERGENS.

1658
01:06:06,487 --> 01:06:09,223
THAT'S NOT HOW I THINK OF IT.

1659
01:06:09,223 --> 01:06:11,358
BUT WHETHER THEY COULD -- IN THE

1660
01:06:11,358 --> 01:06:13,193
WAY THEY CAN TUNE TYPE 2

1661
01:06:13,193 --> 01:06:15,262
IMMUNITY AND DO HAVE A ROLE IN

1662
01:06:15,262 --> 01:06:17,297
FOOD ALLERGY IS WHAT WE'RE

1663
01:06:17,297 --> 01:06:21,134
THINKING ABOUT, WHAT THAT COULD

1664
01:06:21,134 --> 01:06:23,737
BE ANTIGEN INDEPENDENT WAY.

1665
01:06:23,737 --> 01:06:25,672
SO, YEAH, WE'VE TIRED CHOLERA

1666
01:06:25,672 --> 01:06:28,442
TOXIN, DOES NOT EXPAND TUFT

1667
01:06:28,442 --> 01:06:29,343
CELLS, AND THAT PRIMING CAN

1668
01:06:29,343 --> 01:06:32,713
STILL HAPPEN IN THE ABSENCE OF

1669
01:06:32,713 --> 01:06:33,413
TUFT CELLS.

1670
01:06:33,413 --> 01:06:36,984
BUT SOME -- WE HAVE SOME

1671
01:06:36,984 --> 01:06:39,353
PRELIMINARY EVIDENCE THAT IF

1672
01:06:39,353 --> 01:06:40,587
YOU'VE ALREADY ESTABLISHED THE

1673
01:06:40,587 --> 01:06:43,257
ALLERGY, THAT TUFT CELLS CAN

1674
01:06:43,257 --> 01:06:44,224
EXACERBATE IT.

1675
01:06:44,224 --> 01:06:45,425
PERHAPS THROUGH WHAT I'VE

1676
01:06:45,425 --> 01:06:47,027
DESCRIBED THROUGH THIS SIGNALING

1677
01:06:47,027 --> 01:06:51,064
ON THE Th2s OR OTHER

1678
01:06:51,064 --> 01:06:51,832
TARGETS.

1679
01:06:51,832 --> 01:06:52,666
DOES THAT ANSWER EVERYTHING?

1680
01:06:52,666 --> 01:06:53,934
>>  I THINK SO.

1681
01:06:53,934 --> 01:06:56,103
THANK YOU.

1682
01:06:56,103 --> 01:06:58,138
>> ALL RIGHT.

1683
01:06:58,138 --> 01:06:58,539
OKAY. 

1684
01:06:58,539 --> 01:07:04,378
THANK YOU VERY MUCH.

1685
01:07:04,378 --> 01:07:04,711
[APPLAUSE]

1686
01:07:04,711 --> 01:07:07,781
[END OF PROGRAM]

1687
01:07:07,781 --> 01:07:13,820
 

1688
01:07:13,820 --> 01:07:23,864