Tiansciibeu by Ana Sangauala }uly 18, 2u14

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Sliue 24 (fiom T Cell Activation Lectuie)-No Title
!"#$%&#'()*+,0k, I might as well iewaiu the people who aie heie at 8 o'clock in the
moining. uoou foi you, anu get staiteu. 0k, backtiacking a little bit, the T-cell gets
activateu LFA binus to IFAN, which allows the t cell to ioll aiounu on the uenuiitic
cell aims. If the t cell binus to BLA plus peptiue, then the LFA changes confoimation.
The t cells tops iolling. The t cell moves ieceptois to the point of contact. The
uenuiitic cell moves ieceptois to the point of contact. You get many ieceptois, t cell
ieceptois binuing to hla peptiue complexes. Anu that's the piimaiy signal, you have
to have the cell ieceptoi iecognize it because you'ie only going to have 1-S t cells foi
any given infection. Then, you have co-stimulation, uenuiitic cell binus to cu28 on
the t cell. That is a ciucial step. 0nce that happens, the t cell is now activateu. It's
going to uo eveiything that it can uo without any fuithei help, so the t cell staits
changes the expiession of the IL 2 ieceptoi. It staits making a thiiu chain foi the IL2
ieceptoi. This high affinity foim of IL 2 ieceptoi is now on the cell suiface. The same
t cell suiface is secieting IL2, so the IL2 fiom the t cell binus to this high affinity
foim of the t cell ieceptoi. It causes t cell piolifeiation. All of the uaughtei t cells will
have the iuentical ieceptoi to the paient t cell anu that is clonal piolifeiation.

Sliue 2S- T cell Piolifeiation
!"#$%&#'()*+,So we now enu up with an aimy of t cells. They aie iuentical to the
paient t cell.

Sliue 26-Biffeientiation Pathways of CB4 T cells
!"#$%&#'()*+,Anu they stait to uiffeientiate. Biffeientiation is uiiven by cytokines.
So, if the t cell, at the beginning of the iesponse, most of the t cells aie going to be
eithei TB1 oi TB2. TB1 t cells uiffeientiate. You get TB1 t cells if the t cell is bounu
by gamma inteifeion oi IL 12. You get TB2 t cells if the t cell is bounu by IL4. Latei
on in the immune iesponse is when you stait getting iegulatoiy t cells. The function
of a iegulatoiy t cell is to suppiess the activity of TB1 t cells. You want TB1 t cells at
the beginning of the iesponse. You neeu to get them shut uown latei on. So, in that
case, the t cells aie going to uiffeientiate fiom TuF-beta. All of this woiks thiough
seconu messengeis. So binuing to the cytokine ieceptoi activates a ceitain seconu
messengei. Biffeient cytokine ieceptois activate uiffeient seconu messengeis. The
seconu messengeis then go off anu go thiough phosphoiylation events anu
eventually you enu up with something in the nucleus that activates tiansciiption
factois. Biffeient tiansciiption factois tuin on uiffeient sets of genes. It's a cassette.
So, T-bet, which is the tiansciiption factoi that gets tuineu on by binuing thiough IL
12 oi gamma inteifeion tuins on a cassette of cytokine genes that incluue IL 2,
gamma inteifeion, anu some otheis. IL 4, binuing thiough the IL 4 ieceptoi, you
eventually tuin on a uiffeient set of seconu messengeis. You eventually enu up
tuining on the tiansciiption factoi uATA-S. uATA-S then tiansciibes a cassette of
genes that incluue IL 4 anu IL S anu some othei things. Binuing thiough TuF-beta,
you eventually tuin on, you go thiough yet a thiiu seconu messengei system. You
Tiansciibeu by Ana Sangauala }uly 18, 2u14
eventually tuin on Fox PS. Fox PS then tuins on a set of genes that incluue TuF-beta
anu IL 1u. 0k. So binuing thiough the cytokine ieceptois, you tuin on oi
phosphoiylate a seconu messengei. Each gioup of cytokines has a uiffeient set of
seconu messengeis. Similai cytokines, IL 12 anu gamma inteifeion aie pio-
inflammatoiy cytokines. So, the pio-inflammatoiy cytokines shaie a
phosphoiylation set that enus up tuining on pio-inflammatoiy cytokines. The anti-
inflammatoiy cytokines like IL 4 use a uiffeient set of seconu messengeis. They
phosphoiylate a uiffeient set of seconu messengeis. They tuin on uiffeient
tiansciiption factois that give you anti-inflammatoiy cytokines. Al iight.

Sliue 27-Alteinate uCB4+ T Cells
!"#$%&#'()*+,Theie aie also 2 othei kinus of t cells. Theie's a little bit of lack of
claiity. They'ie in the thymus. Natuial t iegulatoiy cells, which aie opposeu to
inuucible t iegulatoiy cells. Inuucible t iegulatoiy cells come fiom a paient CB4 t
cell. These aie also CB 4 t cells, but in the thymus, cells that come into contact with
Bassell's coipuscles become natuial iegulatoiy cells. So, theie is a subset of t cells
that uevelop in the thymus anu those aie nT iegs. Anu then, theie aie also TB 17
cells. These aie also alpha beta CB4+ t cells. Some uata show that TB 17 cells come
fiom this paient CB4 t cell. Some uata show that TB 17 cells uevelop in the thymus.
Right now, Paim is saying, he's not putting them in the coming fiom the paient t
cells, so we will say they aie ueveloping in the thymus. It's not a ciitical point. When
t iegulatoiy cells, IT iegulatoiy cells, anu TB 17 cells come into play is when you
have an infection that you'ie not cleaiing. So the t cells keep piolifeiating. You keep
having TB 1's, you keep having TB 1's, you keep having TB 1's. At that point in time,
since the TB1 t cells aien't getting iiu of the infection, you have high levels of
cytokines anu you stait to uiffeientiate off these othei kinus of cells. The IT iegs,
which then suppiess TB1 function, anu the TB 17 cells anu the cytokines piouuceu
by TB 17 cells can eithei help uisease oi get iiu of uisease uepenuing on which
panel they aie anu we will talk about that touay.

Sliue 28-No Title
!"#$%&#'()*+,So we've got these 2 uiffeient kinus of t cells. So now, we have a whole
bunch of t cells anu wheie aie all of these t cells at this point in time. What oigan.
They'ie in the lymph noue. Wheie is the pathogen. Somewheie else. Yeah, not in the
lymph noue. So we neeu to get TB1 t cells fiom the lymph noue to the site of
infection. The way that happens is calleu lymphocyte tiafficking. So oui oiiginal
lymphocyte, oui nave lymphocytes, they have LFA 1 on theii cell suiface anu they
also have an antigen calleu.I'm going to blank on this.L selectin. 0k. So LFA 1 anu
L selectin anu othei things. But the 2 you know about, LFA 1 is the auhesion
molecule that lets the t cell ioll aiounu on the uenuiitic aims. LFA 1 is also founu on
noimal enuothelium. L selectin is founu on the t cell anu anothei molecule founu on
noimal enuothelium is calleu sialateu Lewis X. Sialateu Lewis X. So the shoit hanu
foi that is SLex. Anu heie it is up heie. Sialateu Luessex. So the L selectin on the
nave t cell binus to the SLex on the unactivateu enuothelium. All nave t cells anu
this is also tiue foi b cells, all nave t cells have L selectin anu unactivateu noimal
enuothelium has S Lex. So that's how t cells, anu I CANS, anu some othei things,
Tiansciibeu by Ana Sangauala }uly 18, 2u14
that's how t cells aie moving along thoiugh the ciiculatoiy system. They ioll, they
always ioll. When the uaughtei t cells stait to uiffeientiate, the TB1 t cells lose L
selectin. When the uaughtei cells stait to uiffeientiate, the TB1 t cells lose L selectin.
Anu they ieplace it with a molecule calleu vLA 4, v foi victoi. vLA 4. Now, vLA 4,
unlike L selectin cannot binu to sialateu Lewis X. So, the t cell cannot go to places
that only have S Lex as a piotein on the cell suiface. The infecteu enuothelium, the
activateu enuothelium at the site of infection has gotten iiu of its S lex anu it's
ieplaceu it with a molecule calleu v CAN, v foi victoi. Theie is a concentiation
giauient of v CAN, so the faithei away you get fiom the site of infection, the less v
CAN theie is on the suiface fo the enuothelium, but theie is still going to be vCAN
that's gotten to the enuothelium all the way up to the lymph noue. The activateu t
cell, because it uoesn't have S lex, it can't go back to the high enuothelial venule,
because that's an S lex, soiiy it uoesn't have L selectin. That's an L selectin, S Lex
inteiaction. It can't uo that so how's the othei way to get out of a lymph noue. The
effeient lymphatic. The effeient lymphatic because this has been activateu is going
to have v CAN so the activateu vB 1 t cells cannot go into the peiipheial ciiculation.
They have to go thiough the effeient lymphatic because that's the molecule that
they have. So they go out into the effeient lymphatic anu as they move back, so
lymphatic all uiain back into the vena cava. So they go out the effeient lymphatic,
they uiain back into the vena cava. Now they'ie going to be able to ioll only along
enuothelium that has v CAN. So the place they'ie going to go is towaius the site of
infection. The closei you get to the site of infection, the moie vCAN theie is. That
will steei them to the coiiect aiteiioles, which then steeis them into the coiiect
capillaiy beus that enu up uiaining iight at the site of infection. Nave B anu T cells,
they still have L selectin, anu the only place they can go is back into the high
enuothelial venule anu back into the peiipheial ciiculation. The activateu t cells,
TB1 t cells, they've lost L selectin. They've gaineu v..vLA4..I'm not talking veiy well
this moining eithei. They've gaineu vLA 4. The only place they can go aie
enuothelium that has vCAN. So the vLA 4 on the activateu t cells takes them out the
effeient lymphatic anu then they follow that concentiation giauient so they enu up
in the capillaiy beus at the site of infection. This ieally lovely system. Activateu cells
go one place. Nave cells stay in the ciiculation.

Sliue 29-No Title
!"#$%&#'()*+, So heie's oui sialateu Lewis X. Beie's oui L selectin. This is
enuothelial venule, this is unactivateu enuothelium.

Sliue Su-Activateu vs Resting T cells have uiffeient Auhesion Nolecules
!"#$%&#'()*+,So oui iesting, unactivateu t cells. They have L selectin. They lose L
selectin anu they uon't have vLA 4. 0ui activateu T cells, they've lost L selectin anu
they've gaineu vLA 4. 0nactivateu enuothelium has S Lex. Activateu enuothelium
loses S Lex anu gains v CAN. So keep the v's togethei as you aie iemembeiing this.
vLA 4 binus to v CAN, so that othei thing must binu to S Lex. 0i the L in L selectin
binus to the L in S Lex. Al iight. So oui activateu TB1 t cells aie now going to
migiate off to the site of infection.

Tiansciibeu by Ana Sangauala }uly 18, 2u14
Sliue S1-No title
!"#$%&#'()*+, LFA 1 anu I CAN aie on APC's. vLA 4, v CAN on activateu
enuothelium.

Sliue S2-Tiafficking
!"#$%&#'()*+,skippeu

Sliue SS-Activateu Th1 cells leave the Lymph Noue
!"#$%&#'()*+, So the activateu t cells, they leave the lymph noue.

Sliue S4- Enu Result
!"#$%&#'()*+, Now, the othei kinu of t cell, main kinu of t cell, the TB2 t cells, they
lose L selectin. They uon't get vLA 4. So TB2 t cells, as they uiffeientiate out, they
lose L selectin, they uon't get vLA 4. Wheie aie the TB2 t cells going to be. They stay
in the lymph noue. So TB1 t cells, CB4+ Th1 t cells, aie the geneials that go off to the
site of infection anu make all the othei cells woik togethei. That's how you get iiu of
extiacellulai pathogens. CB4+ Th2 t cells, they stay in the lymph noue anu theii job
is to get b cells activateu. So the Th1 t cells leave the lymph noue, Th 2 t cells also
lose L selectin but they uon't get anything. In the absence of a molecule that lets
them leave the lymph noue, they'ie stuck theie. So they migiate fiom the paiacoitex
ovei to the geiminal centeis, the piimaiy follicles.

Sliue SS- CB8 T cell activation
!"#$%&#'()*+,Now, we've been talking about CB4 t cells. Foi puiposes of this class,
CB8 t cells aie activateu the same way CB4 t cells aie. Al iight. Foi puiposes of this
class, CB8 t cells aie activateu the same way CB4 t cells aie, except CB8 is binuing to
BLA Class 1 insteau of BLA Class 2. Anu the ieason, we aie simplifying this a bit.
Theie aie actually seveial uiffeient mechanisms that CB8 t cells get activateu anu
one of them is the same way that CB 4 t cells get activateu. It seems to be pathogen
uepenuent. Some pathogens, CB8 t cells can be activateu just fine by themselves.
Some pathogens, you have to have help fiom the CB4 t cells. Foi puiposes of this
class, we'ie not going to woiiy about the othei two. We aie just going to say that
CB8 t cells aie activateu the same way that CB4 t cells aie, except they use BLA Class
1 insteau of BLA Class 2. That finishes T cell activation.

Biu anybouy bothei to ieau the email that I sent out. So you know that confeience is
going to be wheie you guys aie going to tell me the answeis to the stuuy guiue
questions. Anu eveiybouy talks. (Somebouy asks an inauuible question) No they uo
not.

6 7"&& ;0#/)9+#1

Sliue 1 (T Cell Functions Poweipoint)-Review
!"#$%&#'()*+, CB8 T cells, once they'ie activateu, anu we'ie going to stait talking
about this now. CB8 t cells, foi puiposes of this class. Foi puiposes of this class
means that I'm lying but the ieal answei is way moie complicateu than you neeu to
Tiansciibeu by Ana Sangauala }uly 18, 2u14
know as a uentist. You neeu to know what CB8 t cells uo, but the nuances, not so
much. So, foi puiposes of this class, CB8 t cells aie CB 8 t cells, aie CB8 t cells. What
they aie is killing machines. That's all they uo; well, they ielease cytokines. But they
kill things. 0k, so how uo you activate t cells. What's the fiist step. I know it's eaily
in the moining anu it's Fiiuay. I CAN. 0k. What's next. T cell ieceptoi to NBC.
What's next. Elevate titins. What's next. (Inauuible iesponse) 0k. Anu then what
happens. Now you've activateu youi t cell. What's the most impoitant step in that
list. T cell ieceptoi ok. If the t cell ieceptoi uoesn't binu, that t cell isn't going to uo
anything. It's going to go back into the high enuothelial venule anu leave. So TBE
N0ST INP0RTANT STEP is the t cell ieceptoi. If the t cell ieceptoi binus, then you
must have co-stimulation oi you will enu up with an eneigic t cell that's not going to
paiticipate. That's a fail safe. You have to have the t cell ieceptoi binu. If the t cell
ieceptoi uoesn't binu, nothing else neeus to happen. What about uenuiitic cells.
What about uenuiitic cells make them specializeu to activate t cells. Cleaily I wasn't
thinking giammatically when I wiote that. Why aie BC's goou at activating T cells.
(Inauuible iesponse) 0k, say it again. They have both Class 1 anu Class 2, CCL 18,
iight, they seciete the chemokine that ieciuits nave t cells. What else. They can be
infecteu by any viius, anu 2 moie things. (Inauuible iesponse). Paiuon. (Inauuible
Response) 0k. That's tiue but that's not one of the unique things in BC. (Inauuible
iesponse) Paiuon. That's kinua what she saiu, soiiy. 2 Noie. (Inauuible iesponse)
Nope. They can put viiuses in vesicles so they can piesent viial paiticles on both
BLA Class 1 anu BLA Class 2. Anu theie's one moie thing. Paiuon. They uon't get
killeu off because they'ie making inteifeion alpha anu inteifeion beta. Right, so the
uenuiitic cell gets infecteu anu it uoesn't get killeu. It's veiy haiu to activate a t cell if
it's ueau. uoou. Bow uoes tiafficking woik. Nave t cells have L selectin. 0nactivateu
enuothelium- S Lex. Activateu t cells-TB1 anu NCB8. CB8 also uiops L selectin anu
gets vLA 4. 0k. Activateu enuothelium. v CAN. uoou. What aie the uiffeiences
between CB4 anu CB8 t cell activation. 0ne of them uses BLA Class 1 anu one of
them uses BLA Class 2.

Sliue 2-CB4 T Cell Biffeientiation
!"#$%&#'()*+,0k, so T cell uiffeientiation. The Cytokine Nilieu, milieu-the cytokine
enviionment. So the cytokine enviionment, milieu is going to tell you what t cell
you'ie going to get. This is all uiiven thiough seconu messengeis anu ceitain seconu
messengeis give you uiffeient tiansciiption factois. Biffeient tiansciiption factois
activate uiffeient cassettes of genes. That's what makes a TB1 t cell oi a TB2 t cell a
TB2 t cell.

Sliue S-CB4 T Cell Biffeientiation
!"#$%&#'()*+, So, TB1 t cells. The oiiginal binuing is thiough the IL 12 oi the
inteifeion gamma ieceptoi that activates T BET. When you activate T BET, you get a
seconu messengei pathway that causes the tiansciiption of gamma inteifeion anu
the gamma inteifeion ieceptoi. That also is acting thiough seconu messengeis.
Piouuction of gamma inteifeion helps inciease the amount of T BET anu it also
tuins on the tiansciiption factoi STAT 1. That causes an inciease in the IL 12
ieceptoi. That causes moie IL 12 binuing. That gives you moie T BET, which gives
Tiansciibeu by Ana Sangauala }uly 18, 2u14
you moie gamma inteifeion anu gamma inteifeion ieceptoi which gives you moie
STAT 1 anu STAT4 which is a maintenance. T cells woik in loops anu this is a loop.
So t cell ieceptoi binuing anu then the cell staits to piolifeiate anu then it staits to
uiffeientiate. If it gets IL 12, you get T BET. T BET, by getting uiffeient seconu
messengeis. When T BET gets to the nucleus, it staits tiansciibing both gamma
inteifeion anu the gamma inteifeion ieceptoi. Binuing to the gamma inteifeion
thiough a uiffeient set of seconu messengeis then causes STAT 1 tiansciiption
along with TBET tiansciiption. Anu then you get STAT 1, STAT 4, anu T BET anu
that keeps that cell as a TB1 t cell.

Sliue 4-CB4 T Cell Biffeientiation
!"#$%&#'()*+, The oiiginal binuing, t cell ieceptoi binuing, anu you get IL 4 anu that
gives you uATA S thiough seconu messengeis. uATA S gives you moie IL4
piouuction anu IL4 ieceptoi piouuction. IL 4 uses a uiffeient set of ieceptois than
this oiiginal binuing. That causes the tiansciiption factoi STAT 6. STAT 6 then helps
keep tiansciiption of uATA S, which gives you IL 4 anu the IL 4 ieceptoi. Binuing
thiough the IL 4 ieceptoi gives you STAT 6, which gives you moie IL 4 ieceptoi to
give you moie uATA S, which gives you moie IL 4 ieceptoi anu IL 4. So the t cell
secietes the cytokine it neeus to maintain its phenotype.

Sliue S-CB4 T Cell Biffeientiation
!"#$%&#'()*+,CB4 iTiegs, inuicible Tiegs. TuF beta gives you Fox PS. Anu the way
you tell these cells apait eaily on is you uo something calleu intiacellulai flow
cytometiy anu you look to see what tiansciiption factoi is tuineu on. 0nce the cell is
uiffeientiateu out a little bit fuithei, it will have ieceptois foi gamma inteifeion anu
IL 12 wheie it will have ieceptois foi IL 4 anu then you can tell that way. Fox pS
gives you the anti-inflammatoiy cytokines, TuF beta anu IL 1u among othei things.

Sliue 6-Alteinate CB4 T Cell Activation
!"#$%&#'()*+,Natuial T iegulatoiy cells aie geneiateu by Bassell's coipuscles anu
they won't come into play until latei on in an immune iesponse if you'ie not getting
a pathogen cleaieu anu you continue with a heavy uuty t cell iesponse. TB 17 cells, if
you have TuF beta anu IL 6 oi IL 21, you tuin on the tiansciiption factoi R0Rc. That
gives you a uiffeient set of pio-inflammatoiy anu anti-inflammatoiy cytokines. They
aie a family calleu the TB 17 family, not suipiisingly. So, IT iegs, it's TuF beta, you
get Fox pS, TB17 cells, it's TuF beta plus IL 6 oi IL 21. That gives you R0Rc, which
gives you a uiffeient set of pio anu anti inflammatoiy cytokines.

Sliue 7-2
nu
Nessengeis
!"#$%&#'()*+,Now the ieason that I am stiessing seconu messengeis is, anu I'm
going to tell you a little stoiy in this point of time. Autoimmune uiseases, like
inflammatoiy bowel uisease, like Ciohn's uisease is an example, oi iheumatoiu
aithiitis, aie TB1 t cell uiseases. Foi a veiy long time, the only uiugs you coulu use
to tieat these uiseases weie massively immunosuppiessant. You put people on
steioius, you put people on methyltiexate. What these uiugs uiu was pieventeu t
cell activation, peiiou. So, steioius have A) you've completely suppiesseu the
Tiansciibeu by Ana Sangauala }uly 18, 2u14
immune iesponse. If somebouy gets a colu, theie aie no t cells, then you get no b
cells, then you stay sick. Anu both of those uiugs have ieally significant, nasty siue
effects. So about 1u yeais ago, people staiteu making what aie calleu biological.
Eveiybouy has a geneial iuea of what an antibouy is iight. Antibouies aie pait of
oui natuial immune uefense but they'ie also tools. We use antibouies in the
laboiatoiy to iuentify specific pioteins anu to puiify specific pioteins. So we make
antibouies against a ceitain piotein anu that lets us stuuy it. Now we'ie using them
in meuicine, so we've maue antibouies against cytokines. You can have an antibouy
against IL 1 oi an antibouy against TNF alpha oi an antibouy against IL 6. You know
these uiugs. You heie about them on television: Bumeia, anti IL 6 antibouy.
Remicaue, anti IL 1, anu TNF alpha antibouy. So if you give these people, insteau of
just suppiessing all t cells, they staiteu just getting iiu of the excess IL1 oi excess
TNF alpha, oi excess IL 6. By getting iiu of the IL 1, TNF alpha, oi IL 6, you cause the
uisease to gieatly go into iemission so people aie a lot bettei. The siue effects aie
not neaily so significant. You can activate t cells. If you get sick, you'ie bettei at
fenuing off a uisease than people who aie on steioius oi methyltiexate. If you get iiu
of TNF alpha, anu somebouy gets an infection, oi if you have a patient who has
iheumatoiu aithiitis anu they come in iight aftei theii TNF alpha tieatment. You get
in theie anu you uo this ueep anu heavy scaling anu push all this bacteiia into theii
tissue, they can't occluue theii veins. Insteau of a localizeu bacteiemia that goes
away in 24 houis, they get systemic bacteiemia anu they uie. As uentists, you'ie
going to have to be ieally caieful when you tieat patients with these biological
uiseases. 0ne of the big uiawbacks of these is you have to give them as an infusion.
So eveiy 2 months, the peison goes anu sits in the uoctoi's office foi 4 houis anu
they get this antibouy liquiu mixtuie infuseu into theii veins anu they have to hang
out because most of the siue effects aie immeuiate, the nasty siue effects. So they
have to stay theie, so they can make suie that it's not going to uo anything
immeuiate. Anu that's a pain in the butt. It's also, you know, you've alieauy got the
activateu T cells. You'ie getting iiu of the symptoms by getting iiu of the excess
cytokines, but you haven't ieally fixeu the pioblem. So theie is now a new family of
uiugs. What they uo, is they inhibit the seconu messengei }AK that comes thiough
the gamma inteifeion ieceptoi. This is a pill. It's calleu Zoljans. It's a pill that you
take eveiyuay. Now insteau of having the IL 1 secieteu you'ie stopping the t cell
that's going to tiy anu piouuce IL 1 foi making it in the fiist place. So no IL1, no TNF
alpha, no IL6 anu no gamma inteifeion. The ieason you aie leaining this is you aie
now going to have patients who come into youi office taking this uiug. Again, they
aie not going to be able to piouuce the cytokines they neeu to get iiu of heavy uuty
scaling. You'ie going to have to, you know, it's a veiy new uiug. Nobouy has thought
about how this is going to impact uentistiy, so you'ie going to have to think about
this. You may have to have them be off theii uiug foi a couple of uays. A lot of
iheumatoiu aithiitis patients-they have no hanu eye cooiuination. They have
iheumatoiu aithiitis in theii hanus. They've spent yeais on steioius anu steioius aie
ieally hau foi oial tissues. They have teiiible oial hygiene. They finally feel well
enough to come into the uentist anu you'ie like oh my gou, I've got all this calculus
anu bacteiia to get out of theie. Well, if they just hau theii infusion oi if they'ie
taking something that's pieventing them fiom being able to seciete TNF alpha,
Tiansciibeu by Ana Sangauala }uly 18, 2u14
you'ie going to uo some seiious uamage to them. You neeu to be awaie of this. The
ieason you aie leaining this now is theie's a uiug against it.

Sliue 8-IFNy Activateu }AK-STAT
!"#$%&#'()*+, So again, uetails aie not impoitant. So what we have is we have
inteifeion gamma binuing to the inteifeion gamma ieceptoi. It activates }AK. These
}AK's, they tuin on STATS. The STATs then go off anu tuin on tiansciiption of othei
pio-inflammatoiy cytokines. So IL 4 also woiks thiough a }AK-STAT pathway, but
it's a uiffeient set of }AKS. You uon't suppiess TB2 t cells at all. You just suppiess the
TB1 t cells anu they get activateu, they uiffeientiate, but they'ie not able to seciete
the cytokines that they noimally seciete. They can uo theii othei things, but they'ie
not going to seciete theii cytokines.

Sliue 9-}AKSTAT
!"#$%&#'()*+,}ust to give you an example so again uon't memoiize any of this. Know
that each cytokine ieceptoi acts thiough uiffeient }AK-STAT pathways. 0k.

Sliue 1u-Nutual Inhibition
!"#$%&#'()*+, Now, theie's inhibition. So once you stait making TB1 T cells, you
actively pievent that t cell fiom changing it's minu anu becoming a TB2 t cell. So
once you get T BET oi STAT 1 anu gamma inteifeion. 0i gamma inteifeion anu IR
12, you actively pievent uATA S tiansciiption factoi fiom being tuineu on. You
hypeimethylate that BNA. If you get uATA S, that you've gotten thiough IL 4, you
actively pievent T BET fiom being tuineu on by hypeimethylating the BNA. So, TB1
t cells pievent uiffeientiation into TB 2. TB 2 t cells pievent uiffeientiation into TB
1. This is contiast to gamma uelta t cells. uamma uelta t cells can switch back anu
foith. So oiiginally, they usually seciete pio inflammatoiy cytokines, but then they
can stop uoing that anu they can stait secieting anti-inflammatoiy cytokines. Then
they can go back to piouucing pio-inflammatoiy cytokines. CB4 alpha beta t cells
cannot switch. They uo one oi the othei.

Sliue 11-Th1 Effectoi Functions
!"#$%&#'()*+, What uo TB1 t cells uo. They aie at the site of infection. Theii job
fiist of all, the maciophages that aie at the site of infection, they aie able to
phagocytose. They can uo some cleavage of bacteiia that aie in the vesicles oi
viiuses in the vesicles, but that's it. They seciete cytokines, but they'ie not ieally
fully functional. The fiist thing that TB1 t cells uo at the site of infection is they
activate the maciophage to be a fully functional aimeu effectoi maciophage. That
uoes not happen until the TB 1 t cells get to the site of infection. Fully activateu
maciophages then stait secieting ieactive oxygen species. These aie things like
nitiic oxiue anu B2u2, anu the oxygen iauical then pulls ions off of pioteins anu that
causes the pioteins to fall apait. They'ie veiy effective. 0nfoitunately, they'ie veiy
effective on host as well as bacteiia. 0nce the maciophages aie enu stage cells, they
get iampeu up. They uo theii thing foi 2-S uays anu then they iun out of steam. So
once those maciophages aie out of steam, you neeu to get iiu of them. The TB1 t cell
using BLA class 2 on the maciophage, will then binu to the BLA class 2 on that olu
Tiansciibeu by Ana Sangauala }uly 18, 2u14
maciophage anu then kill it. It kills it thiough apoptosis so the contents of the
maciophage get uegiaueu. TB1 t cells cause B cells that get uown to the site of
infection to switch to Igu, which we will talk about on Fiiuay of next week.
Naciophages have ieceptois foi Igu anu they aie much highei affinity than the
ieceptois foi the toll like ieceptois. 0nce the bacteiia is bounu by oi pathogen, I
guess viius too, is bounu by Igu, then the maciophage can phagocytose it much
moie easily. Natuial killei cells have ieceptois foi Igu so once theie is an Igu
attacheu to a pathogen, then the natuial killei cell can kill it. Natuial killei cells uon't
have toll like ieceptois so they can't iecognize pathogens without Igu. It causes
alteiations in BLA class 1. You get moie BLA class 1, anu the pioteasome changes
foimat so that you aie bettei at binuing viial peptiues, cleaving viial peptiues that
will binu to BLA class 1. If it's an intiacellulai pathogen, it's now easiei foi the t
cells, the CB 8 t cells to binu to theii taiget cells anu kill them. Then theie is just an
oveiall inciease in effectoi cell function.

Sliue 12-Why uo you neeu T cells at the site of infection.
!"#$%&#'()*+, So why uo you neeu cells at the site of infection. Beie we see new
maciophages anu these aie olu maciophages. They'ie calleu foamy maciophages
anu they've iun out of juice. These aie the maciophages, wheie if they piesent
peptiue on class 2, those aie the maciophages that get killeu. They can have peptiue
fiom eithei viiuses oi bacteiia because the viial peptiues woulu be bounu by Igu.
The Igu can be iecognizeu by the Igu ieceptoi on the maciophage anu that allows
the maciophage to iecognize the viius. No kinu of antigen piesenting cells, except
foi b cells, have ieceptois foi viiuses.

Sliue 1S-No Title
!"#$%&#'()*+,So the only way uenuiitic cells can iecognize a viius to phagocytose it
is if you put an Igu on it. A uenuiitic cell oi a maciophage oi neutiophil. These aie
active ones again.

Sliue 14-No Title
!"#$%&#'()*+,So heie we see an infection that's going on anu so these bacteiia aie
sitting on top of a maciophage. These bacteiia aie in the maciophage. These aie
sitting on top of fibioblasts. So we have these bacteiia that aie just all ovei the
extiacellulai space anu we neeu to get iiu of them. Anu when we stait binuing Igu
on these bacteiia, it makes it much easiei foi the maciophage to phagocytose them.
You cleai them out.

Sliue 1S- Th1 anu Naciophages
!"#$%&#'()*+, So Th1 t cells anu maciophages. The Th1 t cell binus the maciophage
anu it causes the maciophage to seciete moie cytokines. It causes the maciophage
to change what's in the lysozyme anu you lots of pioteases anu low pB. Then you get
ieactive oxygen species anu you get ieceptois foi Igu. The maciophage befoie it was
secieting cytokines anu it hau lysozymes that coulu cleave with pioteases anu low
pB. Aftei the Th1 t cells, you get moie cytokines. Youi lysozymes auu enzymes to
cleave bacteiia. The maciophages seciete ieactive oxygen species. They put up
Tiansciibeu by Ana Sangauala }uly 18, 2u14
ieceptois foi Igu because now you'ie going to have Igu, which you uiun't have until
you got the Th1 t cell.

Sliue 16-No Title
!"#$%&#'()*+, So heie we see oui Th1 t cell coming to binu the maciophage. This
has to be both a linkeu iecognition so the Th1 t cell ieceptoi has to binu BLA class 2
on the maciophage so it knows it's in an infecteu maciophage. Theie is no point in
activating a maciophage that hasn't eaten bacteiia. Anu CB4u ieceptoi is on the
maciophage anu the CB4u liganu is binuing to the CB4u ieceptoi plus inteifeion
gamma is binuing to the inteifeion gamma ieceptoi is what gives you that fully
functional activateu maciophage that can kill anything in vesicles except
tubeiculosis. It can ielease ieactive oxygen species to uestioy bacteiia that it hasn't
phagocytoseu yet. So we neeu CB4u liganu on the t cell to binu the CB 4u ieceptoi
on the maciophage plus BLA Class 2 peptiue binuing to the t cell ieceptoi. Then the
t cell secietes inteifeion gamma which binus to the inteifeion gamma ieceptoi.
B00N. Naciophages activateu anu it uoes what it uoes ieally well.

Sliue 17-No Title
!"#$%&#'()*+,So heie, the cytokines secieteu by activateu TB1 t cells, we'ie going to
ignoie IL S anu uNCSF because eveiything secietes ILS anu uNCSF so we'ie not
going to woiiy about that. So inteifeion gamma anu CB4u liganu, they get
maciophages going. Fast liganu oi lymphotoxin, that will help kill the chionically
infecteu maciophages. Fast liganu is a membiane bounu cytokine. It kills thiough
apoptosis. IL2 helps suppoit t cell piolifeiation. TNF alpha anu lymphotoxin uo what
TNF alpha anu lymphotoxin uo. They help uestioy enuothelium so you can have
easiei access foi effectoi cells. TNF alpha causes activation of enuothelium anu it
occluues the venules. Anu then, CXCL2 helps chemotaxis. That's chemotactic foi
maciophages. So that helps ieciuit monocytes out of the site of infection. New
monocytes out of the site of infection, they uiffeientiate into maciophages so they
can keep killing the bacteiia as the olu maciophages aie ieauy to uie off.

Sliue 18-TB1 Cytokines
!"#$%&#'()*+,Aliight. So in wiiting, I have a sliue with the function of cytokines so
that means theie aie uetails that you have to know. Anu you will see that theie aie
questions on the stuuy guiue that ask you about the functions of these.

Sliue 19-TB1 Cytokines
!"#$%&#'()*+,CB4u liganu anu, uo I have it. CB4u liganu anu fast liganu aie
membiane bounu cytokines.

Sliue 2u-IFN-y
!"#$%&#'()*+, Bow the t cell iegulates. It's what the TB1 t cell uoes thiough gamma
inteifeion. uamma inteifeion plus CB4u activates the maciophage. It helps inciease
the killing ability in lysozomes anu it also allows the maciophage to seciete ieactive
oxygen species. uamma inteifeion helps the natuial killei cell put up ieceptois foi
Igu. Igu bounu to a bacteiia oi viius can then be killeu off by the natuial killei cell.
Tiansciibeu by Ana Sangauala }uly 18, 2u14
uamma inteifeion causes the epithelia to put up moie BLA class 1 so if the cell is
viially infecteu, it is easiei foi the CB8 t cells to kill the viially infecteu epithelia.
uamma inteifeion causes B cells to isotype switch to Igu. Igu then has ieceptois on
natuial killei cells anu that incieases the efficiency of the maciophage foi
phagocytosis anu it allows the natuial killei cell to have a taiget. Anu then gamma
inteipheion, oops, soiiy. Anu then gamma inteipheion helps make CB8 t cells moie
active. They have to be activateu in the lymph noue by the uenuiitic cell, but at the
site of infection, gamma inteifeion helps keep theii activity high. So gamma
inteifeion is how the TB1 t cell cooiuinates the immune iesponse at the site of
infection. What kinu of t cells get infecteu by BIv. CB4 t cells. So what happens is
you lose the TB1 t cells at the site of infection anu you uon't have any means of
getting youi maciophages going. You uon't have Igu's so the NK cells can't play a
iole. You uon't get activateu CB8 t cells foi most pathogens. Anu so, when you lose
enough CB4 t cells, you lose this. 0nce you lose this, you uie of infections. Because
the only thing that can ieally iun on its own is a neutiophil anu if you'ie amounting
in auaptive immune iesponse, then it means neutiophils weien't uoing enough.

Sliue 21-Not Title
!"#$%&#'()*+,So we'll go thiough this. I'll give you a bieak anu we'll go thiough this
again. So, the maciophage at the site of infection, anu that is the symbol foi
maciophage. It secietes IL 12. IL 12 by itself can activate natuial killei cells but at
this point in time, natuial killei cells uon't have a ieceptoi foi them to be able to
binu bacteiia oi viiuses. IL 12 causes uiffeientiation of TB 1 t cells, of the t cells into
a TB1 t cell. The TB1 t cell then secietes gamma inteifeion. It makes the
maciophage moie active. It causes it to become a fully functional aimeu
maciophage. The TB1 t cell secietes gamma inteifeion that makes NK cells moie
active. NK cells seciete gamma inteifeion secietes gamma inteifeion which helps
keep the TB1 t cells fully functional. The TB1 t cells seciete gamma inteifeion anu
that causes the b cells to isotype switch to Igu. Igu can then binu to pathogens anu
theie aie ieceptois on the natuial killei cells anu theie aie ieceptois on the
maciophages foi the Igu anu that allows the NK cells something to taiget. It allows
the maciophages to be able to phagocytose moie easily. The Igu then binus to the
Igu ieceptoi on the maciophage. The gamma inteifeion secieteu by the natuial
killei cell anu the TB1 t cell then keeps the CB8 t cells moie active. The CB8 t cells
also seciete gamma inteifeion, so the gamma inteifeion then helps keep the
maciophages going anu helps keep the natuial killei cells going anu helps the TB1 t
cells going. So each one of the natuial killei cells anu the TB1 t cells can seciete
moie gamma inteifeion to keep eveiything going. So at the site of infection, this is
what happens. While you've got lots of pathogen theie, this is fantastic. Though, you
aie uamaging host tissue. Those ieactive oxygen species aie uoing a lto of uamage to
host tissue. Tumoi neciosis factoi. 0k. Lymphotoxin is tumoi neciosis factoi beta,
so they'ie causing neciosis on cells that have ieceptois foi them, anu that's host. So
you'ie uoing uamage to host tissue but you aie uoing moie uamage to the pathogen.
0k, so it's a little bit eaily. I will give you 1u minutes now, so we will stait at 8:S6
anu I'll go thiough this loop again.

Tiansciibeu by Ana Sangauala }uly 18, 2u14
Sliue 22-No Title
!"#$%&#'()*+, 0k. Let's go aheau anu get staiteu. So, I've saiu this befoie anu I'll say
it again. Confeience is next week. The stuuy guiues foi innate immunity, antigen
piocessing anu piesentation, t cell uevelopment, anu t cell activation. We'll uo the
othei 4 stuuy guiues foi confeiences the following week. If you'ie in the Nonuay
gioup, the lectuie foi shutting off the immune iesponse follows immeuiately aftei
the confeience. So, go anu listen to last yeai's poucast. That's the one lectuie I'm
ieally not changing fiom last yeai. This is an inteiactive confeience. You uo all the
talking. I uo answei questions, I'll explain things, but I uo not pioviue the answeis
foi the stuuy guiues. Anu EvERYB0BY talks. So I will ask you to aiiange youiselves
into gioups of 4. Nake suie one of youi 4 is somebouy who is somebouy who is
willing to stanu up anu talk because eveiybouy has to talk. Not eveiy peison, but
eveiy gioup has to talk, so nobouy is alloweu to just sit theie anu copy things uown.
The moie woik you put into this at the fiont enu, the moie you will have out of it at
the back enu. The bettei you will unueistanu immunology, which is going to help
you thiough the iest of youi couises anu youi caieei. Also, leaining to think this
way is ciitical to becoming a goou uiffeiential uiagnostician. It's the same piocess
foi figuiing out how this woiks that you use to figuie out uisease. So, oui
maciophage at the site of infection at this point in time is only paitially activateu. It
is secieting cytokines, one of which is IL 12. IL 12 by itself can activate an NK cell,
although at this point in time, the NK cell uoesn't have any taigets. IL 12 also causes
CB4 t cells to uiffeientiate into TB1 t cells. TB1 t cells then stait secieting inteifeion
gamma, which in conjunction with CB4u liganu, activates the maciophage fully. The
aimeu effectoi maciophage is now able to iapiuly lyse anything in a lysosome anu
it's secieting ieactive oxygen species. uamma inteifeion also helps keep the NK cell
active. Active NK cells also seciete gamma inteifeion. uamma inteifeion fiom the
NK cell uoes all the same things that the gamma inteifeion fiom the TB1 t cell uoes.
uamma inteifeion causes B cells to isotype switch to Igu. Theie aie ieceptois on
both maciophages anu NK cells foi Igu anu this is when NK cells get a taiget. They
can binu to whatevei, they binu to the Igu, anu they can kill whatevei is bounu to
the Igu that's bounu to the NK cell. uamma inteifeion secieteu by whatevei cell is
secieting gamma inteifeion also helps alieauy activateu CB8 t cells maintain theii
phenotype. CB8 t cells also seciete gamma inteifeion, which will uo all of the same
things that gamma inteifeion uoes iegaiuless of wheie it is secieteu by. Somebouy
saiu why uo you have all these cells secieting gamma inteifeion. The answei is that
ieuunuancy is goou. So, at the site of infection, you have lots anu lots anu lots of
gamma inteifeion anu gamma inteifeion makes all of the cells play togethei. That
gives you a veiy effective means of uestioying extiacellulai pathogens. You notice
that the TB1 t cell is not binuing to the bacteiia. 0k. TB1 t cells B0 N0T iecognize
bacteiia. TB1 t cells uo not iecognize viius. TB1 t cells uo absolutely nothing
uiiectly towaius a pathogen. They act thiough gamma inteifeion, making othei cells
uo things to the pathogen. B cells can iecognize both bacteiia anu viiuses. The b cell
that's activateu. The b cell secieting antibouy, the antibouy can binu anu that can be
useu as a ieceptoi foi opsonization oi antibouy uepenuent cytotoxicity, which will
talk about Fiiuay, next week. But CB4 t cells uo not uo anything uiiectly to a
Tiansciibeu by Ana Sangauala }uly 18, 2u14
pathogen. Not TB1's, not TB2's, not T iegs, not TB 17's. They act thiough cytokines.
CB8 t cells kill things.

Sliue 2S-Tieg Function
!"#$%&#'()*+,So T iegulatoiy cells. They suppiess conventional TB1 t cells.
Inteiestingly, eveiybouy thought, well if you suppiess TB1, like if you get iiu of T
iegulatoiy cells anu you uon't suppiess TB 1 t cells, uisease is going to get a lot
woise. Some uiseases uo, but some uiseases happen when you get TB1 iegulatoiy
cells out of the pictuie. So, T iegulatoiy cells can both suppiess things anu they can
pievent uisease. So they can pievent some kinus of uiseases anu they actually help
othei kinus of uiseases. So N T iegs aie a pait of thymic toleiance. iTiegs help
pievent peiipheial toleiance. If you have TB1 t cells oi CB8 t cells that aie active
against things, the iTiegs help uowniegulate the function of those TB1 t cells so they
uon't uo too much uamage.

Sliue 24-No Title
!"#$%&#'()*+,So T iegulatoiy cells that aie bounu to an APC will then pievent the
activation of the CB4 t cell. So once you've got an immune iesponse that's going on
long enough anu you uon't want to ieciuit new TB1 t cells into the system, the nT
iegs anu the iTiegs pievent that.

Sliue 2S-Th17 Function
!"#$%&#'()*+,Th17 seciete a bunch of cytokines. IL 17, not suipiisingly. IL 21, IL 22,
anu IL 2S. Notice that not all of them aie pio-inflammatoiy. So IL 22 is anti-
inflammatoiy. IL 21 can go eithei way uepenuing on the uisease.

Sliue 26-Th17 Effectoi functions
!"#$%&#'()*+,Biseases that aie involveu with TB17. So IL 17 is veiy goou against
fungal infections anu it happens when you have a massive inflammatoiy iesponse.
Anu again, when you get TB 17 cells into the mix, you hau TB 1 anu TB 2 t cells.
They've been uoing theii thing anu they can't get iiu of the pathogen. You have high
levels of TuF beta because you have TB2 cells, but you have high levels of IL 6
because the TB 1 cells aie making the maciophages piouuce lots of IL 6. So insteau
of getting an inuucible T iegulatoiy cell, you stait making Th 17 cells. S0 since the
oiiginal cytokine secieteu by the TB1 t cells: the gamma inteifeion, TNF alpha, TNF
beta, CB 4u liganu, fast liganu. Those aien't getting iiu of the pathogen so we'ie
going to tiy a new set of cytokines. That's the Th17 family. These aie all similai to
each othei in stiuctuie anu ielative function. So IL 2S, if you have IL 2S, you have
inflammatoiy bowel uisease anu fungal infections.

Sliue 27- No Title
!"#$%&#'()*+,So uisease inuucing psoiiasis. Some of the cytokines that cause
psoiiasis, which is an autoimmune uisease: IL 17, IL 22, IL 2S. Rheumatoiu aithiitis,
you see a lot of IL 17, IL 21, anu IL 2S. Nultiple scleiosis, you see IL 17 anu IL 2S.
Asthma, we uon't know what's going on theie but Th 17 cells aie playing a iole.
Inflammatoiy bowel uiseases, you see IL 2S. Theie aie some uata that suggest that
Tiansciibeu by Ana Sangauala }uly 18, 2u14
some people's peiiouontal uisease has a TB 17 cytokine family component anu I
think Bi. Ciaig will talk about that. But we have uisease piotective. So we can have
uisease causing IL 2S, but IL 17 anu IL 22 actually piotect against inflammatoiy
bowel uisease. Anu IL 17, IL 21, IL 22, anu IL 2S help the host uefense against
massive infection. So uepenuing on the ielative levels of the uiffeient cytokines, you
can make things bettei oi you can make things woise.

Sliue 28-CB8 T cell function
!"#$%&#'()*+,So what uo CB8 t cells uo. CB8 t cells aie killing machines anu that's
all that they uo. They aie uiiectly lytic to cells containing intiacellulai pathogens. So
CB4 t cells uo not uo anything uiiectly to pathogen. CB8 t cells uo not uo anything
BIRECTLY to pathogen, but CB8 t cells binu to BLA class 1. Any cell that is infecteu
by a viius is going to have viial peptiues on BLA class 1. CB8 t cells, that's what they
uo. They binu to BLA class 1 anu once the t cell is activateu, it uoesn't neeu any of
those othei co-stimulatoiy molecules. It just neeus to binu to BLA class 1. So viially
infecteu cells have lots of viial peptiue on BLA class 1, the CB8 t cell binus to the
viially infecteu cell anu then if the infecteu cell has Fas on it, CB8 t cells can kill
thiough Fas liganu. If the infecteu cell uoesn't have Fas, it can kill thiough an
enzyme system calleu peifoiin anu gianzymes.

Sliue 29-No Title
!"#$%&#'()*+,So cytotoxic t cell, it binus to BLA Class 1. 0k. It moves all of it's lytic
gianules ovei to the point of contact. Anu so, unlike ieactive oxygen species that kill
whatevei they fall on. CB8 t cell ieleases the peifoiin gianzymes uiiectly onto the
suiface of the infecteu cells, so only the infecteu cell gets killeu. So it's highly
specific. But iemembei, what species is the infecteu cell fiom. Bost. Yeah, so CB8 t
cells aie killing Y00. Anu actually, when you have a colu, you all know that you aie
actually most infectious the uay befoie you know you'ie sick. So the symptoms of a
colu aie not fiom the viius. The symptoms fiom a colu aie fiom when youi CB8 t
cells show up at the site of infection anu stait killing off all of youi viially infecteu
cells. Anu then they aie secieting cytokines, IL1, TNF alpha. That's what causes the
fevei. 0nce you lose the integiity of the epithelium, that's wheie all the mucous
comes fiom because theie's nothing to holu it back. So the symptoms of a colu aie
when youi CB8 t cells aie theie. So you've been sick foi 4 oi S uays because that's
how long it takes to get youi t cells activateu, 4 oi S uays. So you've got all of these
viially infecteu cells in youi nasal epithelium. Anu the t cells come along anu they
stait killing off all the viially infecteu cells anu you lose integiity of youi nasal
epithelium. That's wheie all the mucous comes fiom anu then the cytokines aie
what aie secieteu by the CB8 t cells aie what cause fevei. So the fiist 2 uays you feel
ciappy, that's aftei the immune iesponse staits. 0k. Anu a little asiue, gieen tea
activates youi natuial killei cells to seciete inteifeion alpha oi inteifeion beta. So
the minute you stait feeling that sciatchy feeling in youi thioat, stait uiinking lots
anu lots of gieen tea. Zinc pievents viial ieplication, so take zinc tablets, uon't use
the nasal spiay. Anu the best thing you can uo is stay in beu. If you sleep foi the fiist
24 houis oi iest foi the fiist 24 houis, the uuiation of youi colu will be consiueiably
Tiansciibeu by Ana Sangauala }uly 18, 2u14
shoitei than if you show up at school anu infect eveiybouy aiounu you anu tiy to
stuuy. uieen tea, it will make a huge uiffeience in the uuiation of the colu, ok.

Sliue Su-No Title
!"#$%&#'()*+, So heie is my CB8 t cell. It's binuing to peptiue on BLA class 1. It
inuuces apoptosis in that cell anu moves on to the next cell. In fact, it takes 2u
minutes of contact with the CB8 t cell to inuuce apoptosis in the cell. The apoptosis
occuis moie slowly, but 2u minutes of contact with a CB8 t cell, that cell will uie. It
moves to the next cell, 2u minutes, anu that cell will uie. It moves to the next cell, 2u
minutes, anu that cell will uie. CB8 t cells, anu iemembei, it's gone thiough clonal
piolifeiation just like that CB4 t cell, so you've got 1u,uuu CB8 t cells at the site of
infection anu they kill. Anu they kill. Anu they kill. That's all they uo. So one of the
ieasons it's so impoitant to make suie that you, iemembei what cells have BLA
class 1. All of youi nucleateu cells. So if you get aimeu effectoi CB8 t cells activateu
against a host peptiue, those CB8 t cells will show up at the site of wheievei that
tissue is anu they'ie going to kill anu kill anu kill. It's incieuibly impoitant to keep
youi CB8 t cells iegulateu, because once you have them, they'ie going to kill. That's
actually the methou in spinuloaithoiopathies. The CB8 t cells get activateu to a
peptiue on BLA B 27. It's eithei fiom salmonella oi shigella. That peptiue is one
amino aciu away fiom a host peptiue founu on caitilage, heait muscle, enuothelium.
0nce you get the aimy of CB8 t cells, they kill, except now they'ie killing you. So if
you'ie BLA B 27 positive, uon't get foou poisoning.

Sliue S1-No Title
!"#$%&#'()*+,So CB8 t cells, they kill viially infecteu cells. So CB8 t cell is the only
thing that can kill a viially infecteu cell except an NK cell, anu we'ie not going to
woiiy about that. CB8 t cells aie the only things that can kill a viially infecteu cell.
0nce the viius has been ieleaseu into the extiacellulai space, because one of the
mechanism of viial ieplication is that they cause lysis of the infecteu cell anu then
that spieaus out Su,uuu new copies of the viius. 0nce the viius is in the extiacellulai
space, the only thing that can iecognize it is an antibouy. That's the ieason that
when they look at viial uiseases, they want to see how many antibouies you have. If
you have antibouies against the viius, you'ie not going to be ieinfecteu. If youi
antibouy titeis have uioppeu uown too fai, anu you come acioss measles, mumps,
oi chicken pox, then you will get ieinfecteu. In oiuei to get an active immune
iesponse against a viius, you have to have CB8 t cells, but you have to have TB2 t
cells in oiuei to get activateu B cells. Wheie to uo TB2 t cells come fiom. What kinu
aie they. (inauuible iesponse) Say it louu. (Inauuible iesponse) CB4. Right. In oiuei
to effectively get iiu of a viius, you have to have BLA class 1 piesentation foi the
CB8 t cells, but you have to have BLA class 2 piesentation to get youi CB4 t cells
activateu. The TB1's uon't uo a lot. But you have to have TB2's to get antibouies. So
an inteicellulai pathogen iequiies CB4 t cells, it iequiies TB2. The TB2 aim of CB4 t
cells. It iequiies b cells to piouuce antibouy, anu it iequiies CB8 t cells. An
extiacellulai pathogen that's only piesenting on BLA class 2, CB8 t cells nevei get
activateu because they have no iole in this uisease. TB2 t cells-theii job is to activate
b cells anu we will talk about that next Fiiuay aftei we've uone b cell uevelopment.
Tiansciibeu by Ana Sangauala }uly 18, 2u14
Wheie uoes the viius get maue. Insiue the cell. If it's being maue insiue the host cell,
what aie the pioteins going to look like. Bost. Bo maciophages anu neutiophils
have ieceptois foi host pioteins. No. Bo you want them to have ieceptois foi host.
No, you uon't want them to. So what can iecognize that viius when it's in the
extiacellulai space. (inauuible iesponse) Nope. They iecognize peptiue anu BLA
Class 1. So what can iecognize the viius when it's in the extiacellulai space. }ust
antibouies. Wheie uo antibouies come fiom. B cells. Bow uo you get b cells
activateu. TB2 t cells. So, if you just have CB8 t cells, CB8 t cells they kill a viially
infecteu cell, they kill a viially infecteu cell, they kill a viially infecteu cell. The cell, S
cells uown, now iuptuies. It's lyseu by the viius anu Su,uuu copies of the viius come
out anu they can all go out anu infect Su,uuu new cells. So the CB8 t cells can kill the
viially infecteu cell, but how uo you stop the viially infecteu cells fiom infecting new
cells if it iuptuies befoie the CB8 t cell gets theie. So theie's no way to stop new
viial infection with just CB8 t cells. You have to have something to pievent the viius
fiom binuing anu infecting new cells anu that something is an.ANTIB0BY. S0 you
have to have antibouies to get iiu of viiuses. To pievent viiuses fiom infecting anu
it's antibouies that give you immunity, seconuaiy immunity. It's having antibouies in
youi system. You come home, anu youi kiu is coveieu in ieu spots because they
have measles. Well, the ieason you'ie not going to get measles again is because you
have CB8 memoiy t cells that aie going to kill off viially infecteu cells anu you have
antibouies that will pievent viial infection fiom happening. So it's the memoiy fiom
that that keeps you fiom getting viial uiseases. Without antibouies, you get the viial
uisease again.

Sliue S2-yo T lymphocytes
!"#$%&#'()*+, uamma uelta T cells live in the tissue. They aie antigen specific. Al
iight. So they aie antigen specific. So they aie pait of the auaptive immune
iesponse. They live in the tissues. Theii gamma uelta t cell ieceptoi is veiy flexible
like an antibouy, b cell ieceptoi. They iecognize native antigens like b cell ieceptois,
so you uon't have to have BLA class 1. You uon't neeu antigen piesenting cells. They
iesponu to native antigens, anu so the minute you have a viius oi a bacteiia, the
gamma uelta t cell ieceptoi can binu to that native stiuctuie. The gamma uelta t
cells then stait secieting cytokines. At the veiy beginning of the innate immune
iesponse, you also have auaptive gamma uelta t cells anu they help shape whethei
you aie going to have piimaiily a TB1 t cell iesponse oi a TB2 iesponse by the
cytokines they seciete. 0vei time the oiiginal CB4 t cell, oiiginally you stait out with
kinu of equal numbeis of TB1 anu TB2. 0vei time, you'ie going to polaiize one way
oi the othei. If you polaiize towaiu the TB2 iesponse, you'ie going have a lot of
antibouy anu not a lot of cell meuiateu immunity. If you polaiize towaius the TB1
iesponse, you'ie going to have a lot of cell meuiateu immunity, anu not so much
antibouy. Foi many pathogens, it uoesn't mattei which way you polaiize. Foi some
pathogens, it's ciitical. Nycobacteiium tubeiculosis, which causes tubeiculosis: of
you polaiize towaius TB1, so mycobacteiium aie obligate intiacellulai pathogens. If
you polaiize towaiu TB1, you keep the pathogen in check. If you polaiize towaius
TB2, so you have BLA Class 1, anu you get CB8 T cell iesponses, anu you keep the
infection fiom spieauing. If you polaiize towaius TB2 anu piouuce antibouies, well
Tiansciibeu by Ana Sangauala }uly 18, 2u14
antibouies can only fight the mycobacteiium when they hop out of the cell to hop
into the next cell. You uie ieally quickly. The same is tiue in Tb. Nyciobacteiium
tubeiculosis, if you polaiize towaius TB1, you wall off infecteu maciophages with a
gianuloma anu you keep the uisease in check. If you polaiize towaius TB2,
mycobacteiium can only live in cells anu antibouies can't go into cells. You get ieally
seveie uisease ieally quickly. Foi many pathogens, it uoesn't ieally mattei.

Sliue SS-No Title
!"#$%&#'()*+,So, I'm in tiouble heie because I have all of this time to show you
movies that aien't woiking.

Sliue S4-Extiacellulai Pathogens (this section is a lot of Q&A, anu I coulun't heai the
stuuent iesponses to Bi. NcCutcheon's questions)
!"#$%&#'()*+, Except, at the beginning of the lectuie when nobouy's heie. So foi
extiacellulai pathogens. What aie involveu in getting iiu on an extiacellulai
iesponse. Stait at the beginning. 0k, that's not the fiist thing. Complement.
Complement's the fiist thing, ok. 0nce complement is theie, what happens. They
ieciuit neutiophils. 0k. What uo neutiophils uo. They uo phagocytosis anu they can
yeah. What piece of complement uo we neeu. CSB. What uoes that uo immeuiately.
It's a loop so you can get lots anu lots anu lots of complement that leaus to NAC.
What else uoes it uo. It's an opsonin, so the ieceptoi on the maciophage foi CSB has
a much highei affinity than toll ieceptois so the bacteiia stays attacheu to the
maciophage suiface a little bit longei anu that makes it easiei to uo phagocytosis.
The othei thing that complement uoes that we will talk about on Fiiuay is it
activates b cells. What else. We've got neutiophils, maciophages anu complement.
What else. Yup. So, what uo they uo. They change the enuothelium. Al iight. We aie
losing S Lex anu ieplacing it with what. v CAN. Anu then they make it easiei foi
cells to have access anu that also puts CXCL8 on the epithelium so when the
maciophages come to the site of infection, the CXCL8 binuing to the CXCL8 ieceptoi
on the enuothelium is what causes the iolling to stop so that uiapeuesis can occui.
Naciophages anu neutiophils. What's the next step. That's the innate immune cell
pait of the innate iesponse, but you also have auaptive cells: CBS b cells secieting
IgN anu gamma uelta t cells secieting whatevei they'ie going to seciete. So even at
the beginning of the innate immune iesponse, theie aie auaptive cells. That's
anothei ieason that's not a goou uichotomy. So now what happens. (Inauuible
iesponse) It's not maciophages, it's uenuiitic cells. Naciophages aie going to stay at
the site of infection. Benuiitic cells phagocytose the extiacellulai pathogen. Now
what happens. (Inauuible iesponse) You skippeu a bunch of steps, skippeu a bunch
of steps. They have to change theii shape so the uenuiitic cell goes fiom being an
immatuie highly phagocytic cell to a matuie non-phagocytic cell. It has to pull its
aims in so it can get out of the tissues. Then, it uoes what. (Inauuible iesponse) Bow
uiu the NBC Class 2 get theie. (Inauuible iesponse) So you have to tell me how uiu
the NBC class 2 get theie. You've got it up to the vesicle. Bow uiu it uo that. What
happeneu befoie it got to the vesicle. Synthesis on the iibosomes. Then, something
blocks something. Invaiiant chain blocks peptiue binuing gioove. You'ie still in the
ER. It goes to the uolgi, gets put into the vesicle. While it's in the vesicle what
Tiansciibeu by Ana Sangauala }uly 18, 2u14
happens. The outsiue pait of the invaiiant chain gets uegiaueu leaving CLIP. 0k.
The vesicle containing the BLA Class 2 uoes not go into the membiane. Why not.
Because the invaiiant chain is steeiing it towaius othei vesicles. When the vesicle
containing the Class 2 binus to the vesicle containing the bacteiial peptiues, BLA BN
iemoves CLIP, bacteiial peptiue binus the BLA Class 2, anu BLA Class 2 enus up on
the cell suiface. 0k. Now, you can piesent to the t cell. Along with copies of peptiue
bonuing to BLA Class 2, what else is on the cell suiface of the uenuiitic cell. I CAN.
Paiuon. B7. 0ne moie piotein anu something secieteu. CCL18 anu that's going to
attiact what kinu of cells. Nave t cells. 0ne moie ieceptoi. BC sign. BC sign is going
to play a iole in B cell activation. 0k. Wheie is the uenuiitic cell at this point in
time. In the neaiest lymph noue. What pait. Paiacoitex. 0k. So now, the nave CB4
t cells anu actually nave CB8 t cells aie going to come check out the uenuiitic cell.
What will happen to the CB8 t cells. (inauuible iesponse) 0k, so LFA will binu ICAN.
Then what will happen to the CB8 t cells. What kinu of peptiues uoes the uenuiitic
cell have on BLA Class 1. They have host peptiues. What will happen to the CB8 t
cells. N0TBINu. It will finu nothing to binu anu it will go away. What will happen in
99.99999% of the CB4 t cells. Theie will be nothing to binu anu they will go away.
What will happen in that one t cell. T cell ieceptoi will binu to BLA peptiue. Then
what happens. (inauuible iesponse) 0k. Yes. But that's what t cells uo. T cells aie
like teenageis in small towns with uiive ins. They ciuise the stiip, they pull into the
uiive in, they get theii soua, they go back out, they ciuise the stiip, they pull back in,
they get theii Fiench fiies, they go back out, anu they ciuise the stiip. You know,
that's how nave t cells woik. They go into a lymph noue, theie's nothing theie, the
leave the lymph noue. The go into the next lymph noue, theie's nothing theie; they
go back into the ciiculation. They go into the next lymph noue. That's what nave t
cells anu b cells aie uoing. The uenuiitic cell stays in the uiaining lymph noue anu
lets all of the t cells come to it. So, LFA titins. (inauuible question) Yup, which uoesn't
uo us any goou foi infections, so what happens. (inauuible iesponse) Right, theie's
binuing of multiple ieceptois to multiple copies of pathogen. Then, CB28 binuing to
B7, you get t cell activation. What happens. We haven't gotten that fai yet. We only
have one t cell at this point. You can just shout out, you uon't have to iaise youi
hanu. (inauuible iesponse) Bow. We've alieauy uone that. What about IL2. Wheie
uiu the IL2 come fiom. The t cell, the activateu t cell now changes the confiimation
of the IL2 ieceptoi. It causes secietion of IL2. IL2 binus to the IL2 ieceptoi. Now the
t cell, you get piolifeiation. You get this aimy, 1u,uuu uaughtei t cells. What
happens. (inauuible iesponse) Not yet. It's not activateu. Activation is having that t
cell put out IL2 ieceptois anu piolifeiate. Activation means that the t cell is the one
that's going to change its IL2 ieceptoi so that when IL2 binus, it can piolifeiate. All
the othei t cells in the lymph noue, anu that's full of them. They have the low affinity
foim of the IL2 ieceptois so IL2 binuing to them just gives them suivival signals.
They uon't uiviue. (Inauuible iesponse). No, we've got this aimy of cells. We gotta
uiffeientiate them. T cell iuns into IL 12 thiough Tbet. Tbet tiansciiption
tiansciibes what things. (inauuible iesponse) Yes anu the inteifeion gamma
ieceptoi, so inteifeion gamma binuing to the inteifeion gamma ieceptoi causes
what. STAT 1 anu moie Tbet. Then eventually you get STAT4 anu the STAT1 anu
the STAT4 cause tiansciiption of a cassette of pio-inflammatoiy cytokines. What
Tiansciibeu by Ana Sangauala }uly 18, 2u14
else is happening on that TB1 t cell. Wheie is it iight now. It's still in the lymph
noue. What else has to happen. (inauuible iesponse) Bow. What pioteins. vCAN
on the t cell, vLA 4 on the t cell, vCAN on the enuothelium. So now the TB1 goes to
the site of infection. Theie it secietes gamma inteifeion, which uoes boat loaus of
things. What happeneu to the cells that ian into IL 4. TB2 thiough uATAS. uATA S
causes tiansciiption of. IL4 anu. Receptois. So IL4 binuing to the IL4 ieceptoi, you
activate STAT 6. That gives you a cassette of anti-inflammatoiy cytokines that we
will talk about on Fiiuay. Wheie aie TB2 t cells. In the lymph noue. Why uiun't they
go off to the site of infection. They lose L selectin anu they uon't get anything else.
0k. So the TB1 t cells at the site of infection aie getting, as soon as the b cells show
up, aie getting the pathogen eliminateu. Bopefully. If not, then we stait pulling in NT
iegs, IT iegs, anu TB17 cells. BEY!! That was an extiacellulai pathogen. Now,
somebouy has just sneezeu in youi face anu you snoit in a boat loau of viius. What
happens. Not quite. What's the fiist thing that happens with that viius. It infects
cells. What cells. (inauuible iesponse) Among othei things. It infects what it's
supposeu to infect anu then it infects uenuiitic cells. Wheie is complement. Except
foi. So complement is not involveu except foi CSB. Bow uiu we get to CSB. CS can
spontaneously cleave. It's always spontaneously cleaving. If it lanus on host cells,
you have the membiane bounu factois that pievent it fiom becoming a conveitase.
If it lanus on a viius, it stays theie. You uon't get the iest of the complement
pathway because the viius isn't big enough but it will help get b cells activateu. So
we've got CS, we've got infecteu uenuiitic cells, anu we've got CS on viial paiticles.
That's one of the ways that the uenuiitic cells can uo miciopinocytosis. They use the
CSB as an opsonin. We can get viius in intiacellulai, we can get viius in vesicles,
we've got viius infecting. If the viius infects, what happens to the coat pioteins as
they'ie being maue. What's the BC uoing so that it uoesn't get killeu. It makes
inteifeion alpha anu beta. Inteifeion alpha anu beta binuing to the inteifeion alpha
anu beta ieceptois stops viial ieplication. So we have some viius, but the uenuiitic
cells uon't get oveiwhelmeu by viius. So now what happens to those viial coat
pioteins in the cytosol. They'ie uegiaueu by the piotease. They'ie pumpeu into the
ER by. TAP(sp.). Class 1, which goes closely to the suiface. Now, the ieason you
have to know TAP is because, this is an asiue: the human heipes viius family, BBv1,
BBv2, so human heipes viius is the oial heipes, the genital heipes that you get.
That viius, one of the pioteins that gets synthesizeu is a piotein that can block TAP.
It plugs up. TAP is a channel anu it plugs the channel, so you can't get peptiues
cleaveu fiom the cytosol into the enuoplasmic ieticulum. In the absence of peptiue
in the enuoplasmic ieticulum, BLA Class 1 uoes not get folueu up. It uoesn't enu up
on the cell suiface. Eventually, that cell loses all of its BLA class 1. Can a CB8 t cell
kill a cell that uoesn't have BLA Class 1. No. So, TAP pievents CB8 t cells fiom killing
viially infecteu cells. That's actually the iole that NK cells have. So natuial killei cells
have ieceptois that aie inhibiteu by the piesence of BLA Class 1. So cells with BLA
class 1 can be killeu by t cells, inhibit killing by natuial killei cells. In cells that have
lost theii BLA class 1, they cannot be killeu by CB8 t cells. Theie's nothing to inhibit
the killing by the natuial killei cell. Natuial killei cells kill when cells have lost BLA
class 1. So, TAP 1 can block the outsiue, the cytosolic siue. 0gh, not TAP 1. BBv 1 can
block the cytosolic siue of TAP. BCNv can block the ER siue of TAP. BBv2 oi S I
Tiansciibeu by Ana Sangauala }uly 18, 2u14
uon't iemembei, makes a piotein that looks almost like the TAP subunit 2. So, when
TAP 1, the subunit 1, binus with the subunit 2, it uoesn't foim a poie. Then you can't
put peptiues into the enuoplasmic ieticulum. BCNv also makes a piotein that looks
a lot like beta 2 micioglobulin. So it binus to the BLA class 1 heavy chain, but it
uoesn't cause the peptiue binuing gioove to foim piopeily. So, it can't put peptiue
on the BLA Class 1 anu it can't get to the cell suiface. All of the heipes family viiuses
uo something to BLA class 1, so that it can't get to the cell suiface. S of them uo
something to TAP. That's the ieason you have to know about that piotein. As fai as I
know, the iest of the chapeione pioteins that aie in BLA Class 1 synthesis aie not
involveu in uisease. So we've got oui peptiue on BLA Class 1 in what cell. Wheie is
the uenuiitic cell. Not at this point. At this point, it stoppeu being phagocytic,
immatuie cell anu it's a non-phagocytic matuie cell. It's eithei tiaveling to the lymph
noue oi it's in the lymph noue. What's it uoing. It's secieting CCL18, that ieciuits in
nave t cells. Wheie else uoes the uenuiitic cell have peptiue. In vesicles. So wheie
woulu that peptiue enu up. BLA Class 2. So we have peptiue on BLA Class 1, peptiue
on BLA Class 2. What kinu of t cells get ieciuiteu by CCL18. Nave CB4 anu CB8.
This time, what will the CB8 t cells be able to uo. Theie will be one of the CB8 t cells,
anu we ieally uon't want moie than one. Theie will be one of the CB8 t cells that will
iecognize the peptiue piesenteu by one of the BLA Class 1 molecules. That t cell will
get activateu. Those t cells aie wheie at this time. They'ie still in the lymph noue. So
what uo we neeu to uo. They neeu to go to the site of infection. Bow uo you suppose
they uo that. They lose L selectin anu they get vLA 4. They go out stiaight out the
effeient lymphatic. Concentiation giauient. The closei they get to the infecteu tissue,
the moie vLA4 theie is. The CB8 t cells get to the site of infection. They come out of
the ciiculation, anu they stait killing things. 0k. So, long ieview. Let me go see if the
movies aie only going to play at the beginning of the..(uoesn't finish sentence).

|vIBE0j
See how much of a suiface aiea is in contact between those two cells.
|vIBE0j
We uon't caie about that.
|viueoj
So uoes that help the people who want to see in SB. Now how uo I get my movie
back. So now if that's going to woik, let's tiy these.
|vIBE0j
You uon't neeu to know the names of those (iefeiiing to alpha, gamma epsilon
chains in the viueo).
|vIBE0j
0k. So that's what happens with the t cell ieceptoi. If you uon't get signaling fiom
CB28, you'ie stuck at that state anu that's eneigy.
|vIBE0j

0k, so in this, what you neeu to know is that it happens thiough phosphoiylation.
The names of all of those things aie not impoitant, al iight. It happeneu thiough
phosphoiylation. You uo not neeu to know the uetails of that. You B0 neeu to know
}AK-STAT. The iest of these, anu you know, immunology is wonueiful. You get into
Tiansciibeu by Ana Sangauala }uly 18, 2u14
something calleu NAP kinase kinase kinase, which phosphoiylates NAP kinase,
which then goes off anu uoes things. None of that stuff at this point in time is useu
foi uiugs. Theie aie no uiugs baseu on the absence of these, oi at least people uon't
live long enough foi them to be boin with uisease fiom the absence of these so you
uon't neeu to know them. Al iight. So we will pick this up with b cell uevelopment
on Weunesuay.