13_14 Immunology Dr. Leitenberg 1/10/14 4:00 5:00 Notetaker #7 ntigen !

!e"ognition #$li%e& 'rom ( ntigen re"ognition &li%e&)o* re+i&e% ,,t-. $li%e /7 0)i& ne1t ,art o' t)e le"ture i& an im,ortant ,art. n% t)at2& kin% o' *)y I &to,,e% )ere3 I *ante% u& all to be 're&) to get to t)i& im,ortant ,art. 4e"au&e *)at I2m going to talk about no* i& going to &et t)e &tage 'or *)at *e2re going to talk about in a "ou,le *eek&. n% I *ant you guy& to t)ink about *)at *e2re going to talk about no* i& in or%er to kin% o' ,re,are your&el' 'or *)at *e2re going to talk about in a "ou,le o' *eek&. $o I2m going to lea+e you *it) an un&ati&'ie% 'eeling. 5e2re 'ir&t going to %e'ine &ome term& again. $o *e alrea%y %e'ine% an antigen. n antigen i& 6u&t &omet)ing t)at bin%& to an antigen re"e,tor. 4ut no* I *ant to talk about an immunogen. $o t)e 6argon *ay t)at ,eo,le t)ink o' an immunogen i& t)at an immunogen i& an antigen3 but it2& an antigen t)at &timulate& an immune re&,on&e. $o i' you t)ink about t)at an antigen "an bin% to an antigen re"e,tor3 but t)ey2re not all going to in%u"e an immune re&,on&e. $o 6u&t t)e 'a"t o' bin%ing i& not o'ten enoug). It nee%& to be in t)e rig)t *ay to allo* t)e lym,)o"yte to generate an immune re&,on&e. $li%e /7 $o *)at t)at mean& i& t)at all immunogen& are antigen&3 but not all antigen& are immunogen&. n% &o one kin% o' e1am,le t)at *e u&e to talk about t)i& are +ery &mall mole"ule& like %rug metabolite&3 metal like ni"kel3 "arbo)y%rate&3 li,i%&. 0)ey2re u&ually "ra,,y at generating an immune re&,on&e. 8+en t)oug) t)ey "an be re"ogni9e% by immune re"e,tor&3 e&,e"ially t)e 4 "ell re"e,tor3 a& I tol% you t)e 4 "ell re"e,tor "an re"ogni9e ,retty mu") e+eryt)ing. 4ut t)e&e are all antigen& but t)ey2re not immunogeni"3 t)ey %on2t generate an immune re&,on&e. n% one *or% t)at *e u&e to re'er to t)e&e u&ually +ery &mall mole"ule& a& Haptens. :ne *ay o' making t)e ;a,ten immunogeni" i& to "ou,le it to a larger mole"ule3 u&ually a ,rotein. $o t)e ;a,ten by it&el' i& a little &mall mole"ule i& antigeni" but not immunogeni"3 t)en *)en you "ou,le it to a larger ,rotein it be"ome& immunogeni". n% *e re'er to t)at "ou,ling t)ing3 *)en you "ou,le a &mall mole"ule ;a,ten to a larger mole"ule a& a ;a,ten<=arrier "on6ugate. $li%e /> $o t)i& i& a kin% o' e1,eriment about )o* to %e'ine *)et)er t)ing& are immunogeni" *)en you immuni9e a rabbit. 5)en you immuni9e t)e rabbit *it) t)i& ;a,ten3 in t)i& "a&e i& a &mall mole"ule "alle% %initro,)enol3 are you going to make ;a,ten &,e"i'i" antibo%ie&? No3 ;a,ten& are "ra,,y at being immunogeni". I' you immuni9e a rabbit *it) a large ,rotein3 in t)i& "a&e 4$ 3 are you going to get ;a,ten &,e"i'i" antibo%ie&? No be"au&e t)ere2& no ;a,ten. $o i' you immuni9e3 an% you "ou,le t)e ;a,ten on to, o' t)e "arrier ,rotein3 you )a+e a ;a,ten "arrier "on6ugate an% you immuni9e t)e rabbit3 are you going to get ;a,ten &,e"i'i" antibo%ie&? @e&. $li%e 30

$o t)i& i& "lini"ally im,ortant in a +ariety o' *ay&. :ne *ay t)at *e2ll talk about in later le"ture&3 more t)an on"e3 i& t)at t)ere are "ertain +a""ine& t)at take a%+antage o' t)i& ,rin"i,le. $o i' you *ant to make an antibo%y re&,on&e to a ,oly&a"")ari%e e,ito,e t)at2& on t)e &ur'a"e o' ;aemo,)ilu& in'luen9a ty,e b #;ib. *)i") i& a ba"teria3 6u&t gi+ing t)e ,oly&a"")ari%e by it&el' %oe&n2t *ork &o *ell. 4ut i' you "ou,le t)at ,oly&a"")ari%e to a ,rotein "arrier you get a goo% immune re&,on&e to t)e ,oly&a"")ari%e an% it *ill ,rote"t you again&t getting &i"k 'rom ;ib. 5e al&o u&e ;a,ten &,e"i'i" antibo%ie& in a *)ole +ariety o' t)ing& t)at are u&e% in "lini"al laboratorie& to mea&ure t)ing& like %rug metabolite& to ,regnan"y te&t& to &ee i' you )a+e beta<;=A. 0)o&e te&t are ma%e by largely taking a%+antage o' t)i& ,)enomenon. $li%e 31 0)e ot)er ,la"e *)ere t)i& "ome& u, i& in "ertain ty,e& o' allergi" re&,on&e&. $o many o' you kno* t)at "ertain ,er"entage& o' ,atient& are allergi" to &ome antibioti"&. B&ually t)e&e %rug& are ;a,ten<like3 t)ey2re &mall mole"ule& t)ey2re not +ery immunogeni" by t)em&el+e&. 4ut t)ey be"ome immunogeni" *)en t)ey )ook on to a "om,le1 *it) "ell& in your bo%y or ,rotein& a&&o"iate% *it) "ell& in your bo%y or ,rotein& in your &erum. 0)at2& ba&i"ally )o* t)e Ceni"illin allergy *ork&. 0)e Ceni"illin ")emi"al "an lat") onto ,rotein& maybe on t)e &ur'a"e o' !4=& or &erum ,rotein& an% t)at "an in%u"e an immune re&,on&e. Slide 32 $o t)e ba&i" ,oint t)at *e learne%3 t)e 'ir&t ,art o' t)i& you guy& *ill kno*. $o 0rue or Dal&e ;a,ten& are large ,rotein&3 &u") a& 4$ ? Dal&e. =arrier& are &mall mole"ule& &u") a& Ceni"illin? l&o Dal&e. 4ut t)e )ar% ,art o' t)i& i& *)at i& t)e biology be)in% t)i& ,)enomenon? 5)y i& it t)at *)en you "ou,le a ;a,ten onto a ,rotein3 it be"ome& immunogeni"? 5)y i& it ne"e&&ary to immuni9e *it) t)e ;a,ten =arrier "on6ugate in or%er to generate antibo%ie& to t)e ;a,ten? n% t)e an&*er to t)i& Eue&tion i& really a "entral t)ing about )o* t)e immune re&,on&e *ork&. $o I *ant you guy& to ,u99le o+er t)i&3 t)ere *ill be "lue& later on in t)i& le"ture3 but *e2re going to %o it in %etail in a later le"ture t)e *eek a'ter ne1t. $li%e 33 $o any Eue&tion& about anyt)ing *e2+e %one &o 'ar? $o ba&i"ally *)at *e2+e "o+ere% i& t)e 4 "ell re"e,tor3 e,ito,e &tru"ture3 an% t)e &,e"i'i"ity. $o *)at I *ant to %o no* i& talk about t)e 0 "ell antigen re"e,tor3 *)i") i& +ery &imilar to t)e 4 "ell antigen re"e,tor. $li%e 34 $o )ere2& t)e T cell receptor rig)t )ere3 it2& a )etero%imer. Fo&t 0 "ell& )a+e an al,)a ")ain an% a beta ")ain. 0)e "artoon &)o*& it like one arm o' t)e antibo%y mole"ule. $o t)e 0 "ell re"e,tor i& monovalent. It )a& one antigen-binding site. 5)erea& t)e 4 "ell re"e,tor i& bi+alent an% )a& t*o antigen bin%ing &ite&.

$li%e 35 :+erall t)e ba&i" ,rin"i,le i& t)e &ame. @ou )a+e t)i& )etero%imer an% t)e +ariable ,ortion u, )ere i& ma%e u, o' a "ombination o' bot) t)e al,)a ")ain an% t)e beta ")ain an% t)i& i& *)at "on'er& antigen &,e"i'i"ity. n% t)en %o*n )ere i& t)e "on&tant region t)at i& le&& +ariable 'rom "ell to "ell. $li%e 3G 0)ere2& two main kinds of T cell antigen receptors. bout >5H t)at "an be 'oun% in t)e bo%y )a+e t)i& al,)a<beta 0 "ell re"e,tor3 &o a )etero%imer o' one al,)a an% one beta ")ain. :t)er "ell& in t)e bo%y )a+e *)at i& "alle% t)e gamma<%elta 0 "ell re"e,tor. n% *e2ll talk more about "ell& t)at e1,re&& t)i& gamma<%elta re"e,tor in a later le"ture but *)at2& &)o*n )ere i& t)at t)i& kin% o' mark& "ell&. =ell& t)at e1,re&& t)i& re"e,tor ten% to )a+e more innate<like 'un"tion. n% t)ey re"ogni9e antigen3 t)ey kin% o' %i&obey t)e rule& I2m about to tell you in a 'e* minute&. $li%e 37 I tol% you t)at t)e B cell receptor recognizes shapes3 re"ogni9e& "on'ormational e,ito,e&. I' you look at *)at e,ito,e& t)ere are on ;en 8gg Ly&o9yme ,rotein you noti"e t)at t)e 4 "ell re"e,tor& "an re"ogni9e &omet)ing o+er )ere3 a %i''erent on may re"ogni9e &omet)ing o+er )ere3 an% a t)ir% one may re"ogni9e &omet)ing o+er )ere. 4ut i' you ma, t)e e,ito,e& t)at 0 "ell& re"ogni9e3 it look& %i''erent. 0)e T cell receptors seem to be recognizing stretches of peptides t)at are ma%e u, o'ten internally in t)e ,rotein &tru"ture. $o in&tea% o' re"ogni9ing &)a,e& an% 'ol%& on t)e out&i%e mole"ule. 0)e 0 "ell re"e,tor re"ogni9e& &)ort &tret")e& o' ,e,ti%e& t)at &ometime& are not ob+iou&ly e1,o&e% to t)e out&i%e en+ironment. $li%e 37 $o t)i& i& kin% o' a "entral t)ing t)at ,eo,le noti"e% a long time ago. 0)e 0 "ell& an% 4 "ell& rea"te% %i''erently to %enature% +&. nati+e ,rotein&. $o i' you take a nati+e ,rotein an% you get a 4 "ell re&,on&e3 but it *oul%n2t ne"e&&arily a"ti+e t)e 0 "ell& t)at )a% been e1,o&e% to t)at ,rotein. 4ut i' you boile% t)at ,rotein t)en it *oul% re+eal e,ito,e& t)at t)e 0 "ell "oul% re"ogni9e. $o *)en you boil it you %enature it. 4ut i' you boile% ,rotein& you o'ten lo&t t)e ability o' 4 "ell& to re"ogni9e. 0)e ot)er t)ing noti"e% *a& t)at antibo%ie& "oul% be generate% again&t non<,rotein antigen& like ,oly&a"")ari%e&3 but non<,rotein& antigen& *oul% not eli"it 0 "ell re&,on&e&. B&ually it nee%e% ,rotein&. $li%e 3> $o t)i& e+i%en"e kin% o' ga+e ri&e to t)i& i%ea t)at 0 "ell& re"ogni9e a 'orm o' antigen t)at i& %i''erent 'rom *)at 4 "ell& re"ogni9e. $li%e 40 0)i& i& a 'amou& e1,eriment. $o i' you took 0 "ell& an% mi1e% t)em *it) antigen3 not)ing )a,,ene%. 4ut i' you took 0 "ell& an% mi1e% t)em *it) antigen an% t)en a%%e% anot)er "ell kno*n a& an antigen<,re&enting "ell3 t)en you )a% 0 "ell a"ti+ation. $o t)i& i& kin% o' e+i%en"e t)at you nee%e% &ome ot)er &te, in or%er to 'a"ilitate 0 "ell re"ognition o' an antigen. $li%e 41 n% t)at2& be"au&e t)e *ay 0 "ell& re"ogni9e antigen i& t)at t)ey %on2t re"ogni9e t)e *)ole antigen. 0)ey re"ogni9e little ,ie"e&3 little &tret")e& o' ,e,ti%e& *it)in t)at ,rotein

antigen. n% t)o&e &tret")e& o' ,e,ti%e& are ,re&ente% by t)i& &mall ,rotein t)at I intro%u"e% earlier in t)e le"ture "alle% Ma or Histocompatibilit! "omple# molecules $MH"%. n% &o o+er )ere on t)e rig)t i& a "artoon o' one o' t)e&e F;= mole"ule& an% it2& 6u&t a ,rotein t)at2& e1,re&&e% on "ell&. $o it2& an integral membrane ,rotein. n% it )a& t)i& little room )ere t)at "an "ontain ,e,ti%e&. $o *)at t)e 0 "ell re"e,tor re"ogni9e& i& an F;= mole"ule ,lu& t)e ,e,ti%e. $o t)at2& blo*n u, in t)i& "artoon. 0)e 0 "ell& i& abo+e3 t)e F;= mole"ule3 t)e antigen ,re&enting mole"ule belo*. n% *)at you &ee i& t)at t)e F;= mole"ule )a& t)i& bro*n t)ing )ere3 an% t)at2& t)e ,e,ti%e. n% t)e 0 "ell re"e,tor3 *)at t)e 0 "ell re"e,tor i& really re"ogni9ing3 t)e e,ito,e it2& re"ogni9ing i& bot) t)e ,e,ti%e an% t)e F;= mole"ule at t)e &ame time. $li%e 4/ $o one *ay o' %e&"ribing t)i& ,)enomenon i& t)e T cell receptor is MH" restricted. It2& re&tri"te% in t)at it only intera"t& *it) ,e,ti%e& t)at are ,re&ente% by t)e F;=. n% a& you2ll &ee in later le"ture&3 it& &,e"i'i"ity i& re&tri"te% by t)e ty,e o' F;= t)at i& e1,re&&ing t)at ,e,ti%e. $o it& &,e"i'i"ity i& %etermine% by bot) t)e ,e,ti%e an% t)e F;= mole"ule. $o t)i& i& +ery %i''erent 'rom 4 "ell re"e,tor3 *)i") re"ogni9e& "on'ormational e,ito,e&3 it %oe&n2t "are about ot)er "ell&3 %oe&n2t "are about t)e F;=. $o t)e 0 "ell re"e,tor re"ogni9e& a ,e,ti%e ,lu& F;=. $li%e 43 $o t)i& ,)enomenon t)at 0 "ell re"e,tor& re"ogni9e antigen a& ,ro"e&&e% &)ort ,e,ti%e& in a&&o"iation *it) F;= mole"ule& e1,lain& a lot o' &tu''. It e1,lain& *)y 0 "ell& re"ogni9e ,rotein& but no ,oly&a"")ari%e&. It al&o e1,lain& *)y %enature% ,rotein& "an be re"ogni9e% by 0 "ell& be"au&e t)ey %on2t "are about t)e tertiary "on'ormation o' t)e ,rotein&3 t)ey "are about t)e little ,ie"e& o' t)e ,rotein& t)at are tra,,e% in&i%e t)e antigen<,re&enting "ell. n% it al&o e1,lain& t)i& nee% 'or ,)y&i"al intera"tion o' t)e 0 "ell *it) t)i& ot)er antigen<,re&enting "ell. n% t)e F;= re&tri"te% &,e"i'i"ity o' t)e re&,on&e. n% &o *e2ll talk more about *)y t)i& i& im,ortant in regular immune re&,on&e& again in later le"ture&. $li%e 44 $o )ere2& t)e "artoon again3 anot)er "artoon t)at I &)o* a lot. $o )ere2& an antigen< ,re&enting "ell. It may take u, ,rotein& 'orm t)e mi"roen+ironment3 ,ro"e&& t)em into ,e,ti%e&3 an% %i&,lay t)em on t)eir "ell &ur'a"e by t)e&e F;= mole"ule& &o t)e 0 "ell t)en "an engage *it) t)e ,e,ti%e an% t)e F;= mole"ule. 0)at2& really )o* 0 "ell re"e,tor re"ognition *ork&. Iue&tion& about t)at? $li%e 45 $o t)i& i& a "on'u&ing t)ing t)at I2m going to +iolate t)e rule& no*. :ne o' t)e t)ing& t)at I 6u&t tol% you about )o* t)i& *ork&. 4ut t)en t)ere are &,e"ial e1"e,tion& to t)e rule. n% one &,e"ial e1"e,tion t)at i& "lini"ally im,ortant i& &omet)ing "alle% &u,erantigen&. Superantigens are antigen& t)at "an a"ti+ate 0 "ell& *it)out being ,re&ente% in t)at *ay *it) t)e F;= ,e,ti%e<bin%ing route. 5)at normally )a,,en& i& t)e 0 "ell re"ogni9e& t)e F;= ,e,ti%e "om,le1. 4ut t)ere2& "ertain antigen& t)at are able to bin% kin% o' to t)e out&i%e ,art o' t)e F;= mole"ule an% t)e out&i%e ,art o' t)e 0 "ell re"e,tor an% 'a"ilitate intera"tion *it)out ,ro"e&&ing. It 'a"ilitate& engagement bet*een t)e 0 "ell an% t)e antigen<,re&enting "ell an% allo*& t)e 0 "ell to get &ignale% an% a"ti+ate%. It really by,a&&e& t)e 'ine &,e"i'i"ity o' t)e 0 "ell antigen re"e,tor. $o t)e&e &u,erantigen& are &u,er be"au&e t)ey bin% to relati+ely "on&er+e% ,art&3 ,art& t)at aren2t +ariable o' t)e 0 "ell antigen re"e,tor. $o in&tea% o' t)e ty,i"al antigen a"ting on one in a )un%re% "ell& in

your bo%y3 t)e&e &u,erantigen& may a"t on 10H o' t)e "ell& in our bo%y. $o i' you imagine *)at t)e "on&eEuen"e& o' t)at *oul% be3 t)ey are ,retty &e+ere. $o *)at t)at mean& i& t)at *)en you are e1,o&e% to a &u,erantigen3 you a"ti+ate a *)ole me&& o' lym,)o"yte& all at on"e3 really Eui"k3 really &trongly. n% t)ey make lot& o' "ytokine&3 an% t)ey get &u,er &i"k. n% t)ere are "ertain +iru&e& an% mo&tly ba"teria t)at make to1in& t)at are &u,erantigen&. n% t)at2& *)y t)e to1in& are to1i"3 be"au&e t)ey a"ti+ate a lot o' t)e&e 0 "ell& all at on"e. 0)ey make t)e 0 "ell& make lot& o' "ytokine&. $o e1am,le& o' t)at are t)ing& t)at "au&e 'oo% ,oi&oning3 or to1i" &)o"k &yn%rome. 0)ere2& anot)er ba"terial in'e"tion t)at in+ol+e& t)e ,ro%u"tion o' t)o&e &u,erantigen&. 5)at t)at mean& i& t)at your getting a"ti+ation o' lot& o' "ell& really Eui"k. n% it i& yet anot)er e1am,le about e+en t)oug) t)e immune &y&tem i& im,ortant in ,rote"ting u& again&t in'e"tiou& %i&ea&e3 a lot o' t)e time& *)en *e &ee ,eo,le get &i"k i& be"au&e t)e immune &y&tem i& "au&ing )uge %amage by a""i%ent. $o t)ere nee%& to be regulatory me")ani&m& t)at kee, t)e immune &y&tem in ")e"k. n% *e2ll "ome ba"k to t)at. $li%e 4G $o let& talk 6u&t a tiny bit about F;= mole"ule&3 t)i& i& a ,re+ie* 'or ne1t *eek. 0)ere2& two main classes of MH" molecules3 F;= "la&& 1 an% F;= "la&& /. 0)e nomen"lature 'or t)i& i& "on'u&ing but )uman&2 F;= mole"ule& are "alle% ;uman Leuko"yte ntigen&. $o *)en you )ear ,eo,le talk about ;L antigen&& H'( is MH" molecules) $li%e 47<4> Skipped) *ead on !our own Slide +, Let2& %o a little Eui9. 0rue or Dal&e &ingle ,rotein may eli"it a re&,on&e 'rom &e+eral %i''erent "lone& o' lym,)o"yte&? 0rue. &ingle ,rotein may )a+e multi,le %i''erent e,ito,e& t)at "an be re"ogni9e% by %i''erent 0 "ell& *it) %i''erent antigen re"e,tor&. 0 "ell e,ito,e& ten% to be linear ,e,ti%e& t)at bin% to t)e F;=? 0rue. 4 "ell e,ito,e& ten% to be non&eEuential an% are 'orme% by tertiary "on'ormation 0rue. ll antigen& "an in%u"e an immune re&,on&e an% are immunogeni" Dal&e. $li%e 50 $o let2& talk about t)e mole"ular biology o' antigen re"e,tor ,rotein %e+elo,ment. $o t)i& i& t)e ,roblem t)at *e )a% be'ore. 0)ere are million& o' lym,)o"yte& *it) %i''erent antigen re"e,tor& but *e only )a+e about 303000 gene&. 4ut )o* "an t)at be "au&e *e all learne% t)at one gene one ,rotein?

$li%e 51 0)e rea&on 'or t)at i& t)i& ,)enomenon t)at2& &,e"ial3 only )a,,en& in lym,)o"yte& "alle% somatic recombination. During lym,)o"yte %e+elo,ment3 t)e lym,)o"yte& kin% o' ")ange t)eir geneti" &tru"ture one at a time. $o t)ere2& %i''erent region& *it)in t)e antigen re"e,tor gene t)at are o'ten re'erre% to a& mini gene&. 0)ere2& %i''erent "la&&e& o' mini gene&. $o t)i& i& a "artoon &tru"ture o' t)e immunoglobulin )ea+y ")ain gene lo"u&. n% *)at you &ee i& t)at t)ere2& t)e&e - region mini genes& . region mini genes& and / region mini genes. J 'or +ariable3 D 'or %i+er&ity3 an% K 'or 6oining. $o you2+e got t)i& immunoglobulin )ea+y ")ain gene& )a,,ening in 4 "ell& in 4 "ell %e+elo,ment. $o *)at )a,,en& %uring 4 "ell %e+elo,ment3 t)e in%i+i%ual 4 "ell "lone &ay&3 (I2m going to u&e t)i& J region mini gene3 maybe t)i& D region mini gene3 an% I2m going to u&e t)i& K region.L n% it2& going to &,li"e out t)e inter+ening DN an% 6oin t)e&e mini gene& toget)er. $o you )a+e t)i& ,re"ur&or gene t)at i& really big3 an% you )a+e %uring %e+elo,ment you )a+e &,li"ing out o' gene &egment& in bet*een to 6oin %i''erent mini gene &egment& toget)er in a +ariable arrangement. n% t)at +ariation allo*& you to generate million& an% million& o' %i''erent antigen re"e,tor&. $li%e 5/ n% &o one *ay *e u&e to %e&"ribe t)i& i& *e2re %e+elo,ing a %i+er&e antigen re"e,tor re,ertoire generate% by &ite<&,e"i'i" rearrangement o' mini gene &egment&. $o 'or t)e immunoglobulin mini gene &egment& you )a+e J2& an% D2& an% K2&. !emember I tol% you at t)e beginning t)at t)i& %e+elo,ment o' an initial re"e,tor re,ertoire i& antigen in%e,en%ent. 0)i& i& )a,,ening ran%omly3 you %on2t kno* *)at you2re going to get in'e"te% *it) in t)e 'uture. $li%e 53 $o )ere2& again t)e &tru"ture o' antigen re"e,tor gene lo"i. $o remember t)at t)e 4 "ell re"e,tor i& ma%e u, o' bot) t)e )ea+y an% lig)t ")ain. It turn& out t)at there0s two different kinds of light chains& a kappa light chain and a lambda light chain3 %i''erent gene& an% %i''erent ")romo&ome&. Dor bot) o' t)em t)oug)3 t)ey bot) )a+e t)e&e +ariable region mini gene& an% K region mini gene&. n% t)e )ea+y ")ain )a& J3 D3 an% K region&. $li%e 54 $o *)at )a,,en& in t)e DN in t)e 4 "ell %uring %e+elo,ment i& t)e %e+elo,ing 4 "ell *ill ,i"k one o' t)e&e J2&3 one o' t)e&e D2&3 one o' t)e&e K2&3 &,li"e out t)e inter+ening DN an% to*ar%& t)e en% o' t)i& %e+elo,mental ,ro"e&& it *ill ")ange it2& &tru"ture to ,ut t)e J2& an% D2& an% K2& go toget)er all in a ro*. $in"e t)ere2& multi,le %i''erent J2&3 D2&3 an% K2& to ")oo&e 'rom3 *)en you multi,ly t)at all out it en%& u, in a 'air amount o' +ariability. $o again t)i& ,ro"e&& i& "alle% $omati" !e"ombination. 0)e *or% &omati" i& im,ortant )ere. It2& &omati" be"au&e you2re ")anging t)e DN &tru"ture o' &omati" "ell&. 0)e&e are not egg "ell& or &,erm "ell&3 t)ey2re not germ "ell&. @ou2re ")anging t)e geneti" &tru"ture o' a normal "ell. 0)at %oe&n2t )a,,en any*)ere el&e. It2& a really &,e"ial t)ing 'or lym,)o"yte&. $li%e 55 Ku&t to really )ammer t)i& in3 t)i& i& )a,,ening at t)e DN le+el. @ou all learne% t)at DN i& tran&"ribe% to !N an% !N i& tran&late% to ,rotein&. n% t)ere i& t)i& t)ing t)at )a,,en& "alle% alternati+e !N &,li"ing3 *)ere you )a+e %i''erent e1on& o' !N 2& &,li"e% toget)er an% you "an make %i''erent ,rotein& t)at *ay. 4ut t)at2& not *)at t)i& i&. 0)i& i& a"tual ")ange& in DN . $o t)e ")ange& in DN o""ur& in t)e&e %e+elo,ing

lym,)o"yte& are in)erite% *)en it "lonally e1,an%&. $o t)e lym,)o"yte )a& a re"e,tor t)at i& %e'ine% by it& DN 3 *)en it ,roli'erate& it make& more o' it&el'3 t)at DN i& "arrie% on to it& %aug)ter "ell&. $li%e 5G $o t)i& i& 6u&t t)e &ame "artoon I &)o*e% you be'ore. @ou get &,li"ing out o' inter+ening DN 3 &o t)i& i& all at t)e DN le+el. $o all *e2+e talke% about &o 'ar3 t)e &tu'' t)at2& )a,,ening at t)e ,art o' t)e gene t)at en"o%e& 'or t)e +ariable region o' t)e ,rotein. 0)i& i& t)e "o%ing antigen re"e,tor &,e"i'i"ity o' t)e immunoglobulin mole"ule&. @ou noti"e t)e blue e1on )ere labeled ") This is the constant region that0s not involved in this process. $o t)e "on&tant region only "ome& into ,lay in t)e !N ,ro"e&&ing ,)a&e o' t)e ,ro"e&& *)ere you get maturation o' t)e !N tran&"ri,t to make t)e mature me&&enger !N t)at2& ultimately tran&late% into ,rotein. $o I kin%a like t)i& "artoon be"au&e it2& ni"ely "olor "oate%. @ou )a+e t)e J2& an% t)e K2& an% t)e "on&tant region an% you "an &ee *)ere t)e line& o' t)e "artoon are a,,roa")ing. @ou "an &ee t)at t)e re%3 green3 an% yello* ,art are all )ere at t)e antigen bin%ing ,ortion o' t)e "ell. $li%e 57 $o t)ere i& a )uge amount o' *ork t)at goe& into trying to un%er&tan% )o* t)i& ,ro"e&& *ork&. n% I2m not going to talk about it in a )uge amount o' %etail. 4ut I am going to &)o* you a little mo+ie t)at *ill go t)roug) t)e +ariou& &te,& t)at are in+ol+e% in &,li"ing t)e&e %i''erent mini gene &egment&. $li%e 57 4e'ore I %o t)at I 6u&t *ant to &ay one ot)er *or%. $o one *ay t)at t)e&e %i''erent mini gene& &egment& 'in% ea") ot)er %uring t)i& &,li"ing ,ro"e&& i& be"au&e t)ere are t)e&e &eEuen"e& "alle% re"ombination &ignal &eEuen"e& t)at 'lank t)e %i''erent mini gene&. n% t)ey 'lank t)em in a re,etiti+e &tru"ture t)at you %on2t nee% to memori9e. $li%e 5> Movie $li%e G0<G1 $o %on2t 'reak out3 I2m going to tell you *)at you nee% to kno*. 0)ere2& &ome im,ortant ,art&. $o t)i& i& 6u&t *)at *a& &)o*n in t)e mo+ie3 t)e %i''erent &te,& o' t)e rearrangement ,ro"e&&. $o let me go t)roug) t)e ,art& o' t)i& t)at I t)ink are im,ortant an% *)y I t)ink t)ey2re im,ortant. So one protein !ou0re going to know what it is are these *(1 proteins& the recombinase activating proteins& *(1 2 and *(1 2. 0)ey2re ab&olutely e&&ential 'or t)e ,ro"e&& to o""ur3 only e1,re&&e% in lym,)o"yte&. n% t)at2& *)y it )a,,en& in lym,)o"yte&. n% t)ey2re only e1,re&&e% %uring lym,)o"yte %e+elo,ment. n% *e2ll talk more about t)e regulation o' t)e e1,re&&ion o' t)e&e ! A ,rotein& *)en *e talk about %e+elo,ment ne1t *eek. $o t)e&e ,rotein& kin% o' %o *)at t)e mo+ie &ay&3 t)ey ni"k t)e DN an% 'a"ilitate 'ol%ing an% "lea+ing an% t)en t)ere are DN re,air t)at o""ur& to ,ut t)e %i''erent ,ie"e& o' t)e gene toget)er. :ne o' t)e im,ortant t)ing& about t)e mo+ie t)at *e 6u&t &a* t)at you mig)t )a+e mi&&e% *a& it talke% about t)e enz!me TdT )o* it a%%& nu"leoti%e& to t)e &i%e& t)at *ill 6oin. I' you t)ink about t)at it2& kin% o' &"ary rig)t?

$o you )a+e t)e&e %i''erent mini gene &egment& t)at are rearrange% in an or%er t)at allo*& you to )a+e a lot o' %i''erent antigen re"e,tor &,e"i'i"ity. 0)at only ,artially e1,lain& )o* you )a+e million& o' %i''erent re"e,tor&. )uge ,art o' t)e %i+er&ity i& be"au&e o' t)e a"ti+ity o' t)e&e 0%0 en9yme& ")e*ing ba"k or a%%ing nu"leoti%e& at t)e 6oining &ite. n% &o it a%%& +ariable number& o' nu"leoti%e& at t)at &ite. $o you get a )uge amount o' %i+er&ity be"au&e o' t)at ,ro"e&&. 0)at2& an im,ortant ,art o' )o* antigen re"e,tor %i+er&ity "ome& about. Do not memorize all the DNA repair enzymes3 t)ey2re not &,e"ial 'or immunologi&t& be"au&e t)ey2re in all "ell&3 you mig)t learn in 4io")emi&try but I2m not going to tea") it to you. I *on2t te&t you on it. :ne t)ing t)at I %o *ant to ,oint out t)at2& going to "ome ba"k in later le"ture& i& *)ere+er t)ere i& t)i& little loo, o' DN t)at *a& a by,ro%u"t o' t)i& ,ro"e&&3 t)at loo, i& not +ariable. 0)e +ariable ,art i& "o%e% by %i''erent mini gene& an% %i''erent nu"leoti%e& t)at are a%%e% %uring t)e 6oining ,ro"e&&. 4ut t)at loo, )a,,en& e+ery &ingle time an% it )ang& out in t)e&e "ell& 'or a 'e* %ay& or *eek& a'ter t)i& )a& )a,,ene%. n% a'ter t)ey go out into t)e ,eri,)ery. $o one *ay t)at ,eo,le "an "ount )o* many ne* "ell& are being ma%e3 ne* 0 "ell& or ne* 4 "ell&3 i& by looking 'or )o* many "ell& )a+e t)i& little loo, o' DN . 0)i& i& a"tually "lini"ally u&e%3 belie+e it or not3 be"au&e you may kno* t)i& alrea%y but *)ene+er &omeone i& born t)ey get a )eel &ti"k. n% t)at ,ie"e o' bloo% goe& to a ,ubli" )ealt) lab an% it i& &"reene% 'or a *)ole bun") o' %i''erent geneti" %i&ea&e. n% one o' t)e&e %i&ea&e& t)at i& &"reene% 'or i& $e+ere =ombine% Immuno%e'i"ien"y $yn%rome. 4e"au&e you *ant to 'in% t)e&e ki%& *it) t)i& &yn%rome early an% 'i1 it. 0)e &ooner you 'in% t)em be'ore t)ey get in'e"te% an% %ie3 t)e better. $o more an% more ,ubli" )ealt) lab& aroun% t)e "ountry are u&ing t)i& te")nology to i%enti'y *)et)er or not t)ere are ne* "ell& or not. $o i' you %on2t 'in% t)e&e little loo,&3 t)at2& a &ign t)at t)ey2re not making 0 "ell& or 4 "ell& an% t)ey nee% to be e+aluate% 'urt)er. 0)e ot)er *ay t)at I talk *it) you about t)i&3 an% *ill be re,eate% in later le"ture3 an% you may not be able to an&*er t)i& no* but you *ill by later ne1t *eek. I' you )a+e a mutation *)ere you )a+e &omeone born t)at %oe&n2t e1,re&& one o' t)e ! A ,rotein&3 *)at %o you t)ink t)e re&ult *oul% be? 0)ey *oul%n2t make 0 "ell& or 4 "ell&M t)ey2% )a+e a $e+ere =ombine% Immuno%e'i"ien"y $yn%rome. 0)ey *oul% )a+e a +ery early blo"k to t)eir immune &y&tem. $o one o' t)e "au&e& o' $e+ere =ombine% Immuno%e'i"ien"y $yn%rome3 not one o' t)e "ommon one&3 but one o' t)e "au&e&. $li%e G/ $o you nee% to remember *)at t)e ! A ,rotein& are3 t)ey are e1,re&&e% only in %e+elo,ing lym,)o"yte& an% t)ey2re e&&ential 'or t)i& gene rearrangement ,ro"e&&. 0%0 i& anot)er ,rotein t)at i& only e1,re&&e% in %e+elo,ing lym,)o"yte&. It2& not ab&olutely e&&ential 'or gene rearrangement ,ro"e&& to o""ur3 but it really a%%& to t)e %i+er&ity. 0)e&e ot)er ,rotein& t)at *ere in t)e mo+ie like DN ,rotein kina&e3 et" are not &,e"ial to immunologi&t& an% are in+ol+e% in normal DN re,air. $li%e G3 I &)o*e% you t)i& &li%e be'ore3 but no* t)at you2+e &een t)e geneti"& o' t)i&3 you "an &ee t)e %i''erent region& *)ere t)e&e mini gene& are &,li"e% toget)er. @ou *oul% imagine t)at t)e&e are t)e ,art& t)at "o%e 'or t)e mo&t +ariable ,ortion o' t)e ,rotein an% t)at2& true. 0)e mo&t +ariable region i& t)e =D!3 region3 an% t)at i& 6u1ta,o&e% by t)e

JDK &egment. $li%e G4 $o t)i& i& *)at *e2ll re'er to a& &omati" re"ombination mat)3 an% I2m not going to go t)roug) all t)i& but it& 6u&t telling you )o* many %i''erent J region )ea+y ")ain gene&3 D region gene&3 K region gene&3 an% *)at you %o i& multi,ly it all toget)er an% t)en you a%% it to t)e +ariability o' ka,,a lamb%a at t)e en% an% a%% 6un"tional %i+er&ity3 you get a lot o' %i''erent ,otential antigen re"e,tor&. $o t)at2& *)y *e )a+e &o many %i''erent lym,)o"yte& *it) %i''erent re"e,tor& to re"ogni9e t)ing& t)at mig)t not e+en be in nature. $li%e G5 $o t)i& i& 6u&t a &ummary o' *)at *e 6u&t &ai%. $o t)e antigen re"e,tor i& lym,)oi% &,e"i'i". $o it )a,,en& in 0 "ell& an% 4 "ell& not in NN "ell&3 NN "ell& %on2t )a+e t)e&e re"e,tor&. 0)e&e re"e,tor& are only on 0 an% 4 "ell lym,)o"yte&. n% t)i& only o""ur& %uring lym,)o"yte %e+elo,ment3 *e2ll talk about t)at ne1t *eek. It reEuire& t)e ! A ,rotein& an% "orrelate& *it) 0%0 e1,re&&ion. $li%e GG :ne o' t)e rea&on& t)i& i& &tu%ie% a lot i& t)at %uring t)i& ,ro"e&& &ometime& t)e "ell& make mi&take&. $o imagine *)at2& going on )ere i& t)e&e "ell& are re6oining %i''erent DN ,ie"e& toget)er 'rom %i''erent ,art& o' t)e ")romo&ome or maybe 'rom ot)er ")romo&ome&. n% a lot o' time& ,eo,le *it) a"ute leukemia& or lym,)oma&3 &o malignan"ie& o' t)e lym,)oi% &y&tem )a+e tran&lo"ation& t)at bring in on"ogene& into t)i& &ite *)ere antigen rearrangement i& o""urring an% *)ere t)e DN i& a"tually being a"ti+ely tran&"ribe%. $o t)i& i& 6u&t an e1am,le o' one o' t)o&e t)ing&. $o t)e immunoglobulin )ea+y ")ain gene lo"u& i& on ")romo&ome 14 an% t)e F@= on"ogene i& on ")romo&ome 73 an% you2ll )ear ,eo,le re'er to lym,)oma& t)at )a+e an 7<14 tran&lo"ation. n% *)at t)at mean& i& you take t)i& ,ie"e o' DN 3 mo+e% it onto t)i& ")romo&ome &o it& un%er "ontrol o' t)e immunoglobulin en)an"er& an% ,romote& &o you get a"ti+e an% %i&regulation tran&"ri,tion o' t)e F@= ,rotein an% t)i& re&ult& in gro*t) %i&regulation an% t)i& in"rea&e& t)e ri&k o' lym,)oma generation. $o t)i& i& &omet)ing t)at i& re,etiti+e t)at )a,,en& a lot in a"ute leukemia& an% lym,)oma& *it) %i''erent on"ogene&. F@= i& a "ommon one3 you2ll learn a lot about t)i& ne1t year. Jerti"al integration. $li%e G7 $o I2+e &ai% all t)i&3 a"ute lym,)oma& an% leukemia& o'ten re,re&ent in t)e arre&te% &tage o' early lym,)oi% %e+elo,ment *)en t)e antigen re"e,tor rearrangement i& going on. n% it2& t)oug)t t)at t)i& make& "ell& +ulnerable to t)e&e tran&lo"ation e+ent& an% on"ogeni" mutation&. $li%e G7 $o *)at *e2+e talke% about to%ay i& really 6u&t t)i&. 4 "ell re"e,tor &tru"ture3 0 "ell re"e,tor &tru"ture3 4 "ell re"e,tor re"ognition3 an% 0 "ell re"e,tor re"ognition. n% you guy& "an t)ink about t)i& your&el+e&3 but you &)oul% be able to li&t 3 "ommon 'eature& o' t)e 4 "ell re"e,tor an% 0 "ell re"e,tor. n% you &)oul% be able to li&t / t)ing& t)at are %i''erent about 4 an% 0 "ell re"e,tor antigen re"ognition. $li%e G> 5e2ll %o t)i& at t)e beginning o' ne1t le"ture.