1

MICk 360 Immuno|ogy Notes

UNI1 1 Immuno|ogy
What |s Immuno|ogy
Lhe sLudy of lmmune sysLem and response Lo lnvadlng paLhogens
lmmune SysLem collecLlon of cells Llssues and molecules LhaL medlaLe reslsLance Lo lnfecLlons
lmmune 8esponse Lhe coordlnaLed reacLlon of Lhese cells and molecules Lo lnfecLlous mlcrobes
Ldward Ienner laLher of lmmunology
lnoculaLed 8 yr old boy wlLh fluld from cowpox and laLer lnfecLed hlm wlLh smallpox 1he boy dld noL develop
smallpox
Smallpox was eradlcaLed by 1977
Immune System
unlque recognlLlon sysLem ln naLure (Self vs nonself)
lmporLanL for healLh and welfare llfelong proLecLlve lmmunlLy Lo relnfecLlon wlLh Lhe same paLhogen (memory)
LocaLlon ulspersed LhroughouL Lhe body
4 Many dlfferenL cells and subsLances prlmary funcLlon ls defense and survelllance




Slgnlflcance
4 8eneflLs
roLecLlon from Mlcroblal lnvaders
LllmlnaLlon of alLered self (cancer)
4 ueLrlmenLal
ulscomforL (lnflammaLlon)
ulsease of Lhe lmmune sysLem (AluS)
ulsease on Lhe lmmune sysLem (18)
ulsease by Lhe lmmune sysLem (allergy and auLolmmune dlsease)
3 Llnes of uefense
4 1
sL
llne of defense Skln mucous membrane (physlcal barrler) (lnnaLe lmmunlLy)
4 2
nd
llne of defense macrophage (lnnaLe lmmunlLy)
4 3
rd
llne of defense 8 cells and 1 cells Pelper 1 cells coordlnaLes everyLhlng (adapLlve lmmunlLy)


ko|e Imp||cat|ons
uefense agalnsL lnfecLlons ueflclencles resulL ln lncreased lnfecLlons
8ecognlze + responds Lo Llssue grafLs and new
proLelns
lmporLanL barrlers Lo Llssue LransplanLaLlon
uefense agalnsL Lumours oLenLlal for lmmunoLherapy of cancer
ueLecLlng any molecule LaboraLory LesLlng used ln cllnlcal medlclne and
research
2


UNI1 2 Innate Adapt|ve Immun|ty
Innate Immun|ty
- naLural (lnnaLe) lmmunlLy ls lnborn unchanglng and nonspeclflc
- 8road range of LargeLs and nonspeclflc reacLlvlLles
- noL lmproved by repeaLed exposure
- lnvolve physlcal chemlcal barrlers and hagocyLosls
- lnvolve soluble facLors and phagocyLlc cells
Iour 8arr|ers of Innate Immun|ty
- Anatom|c (skln and mucous)
4 3 MAIN IN1LkIACLS Skln Cl LracL (lngesL) resplraLory LracL (lnhale)
1ears sweaL conLalns lysozyme Lo aLLack cell wall (pepLldoglycan) of bacLerla
Cllla/halrs
4 LplLhella produce pepLldes LhaL have naLural anLlbloLlc funcLlon
efens|ns (cysLelnerlch pepLldes presenL ln skln and abundanL ln neuLrophll granules 8eleased
from neuLrophll granules)
uefensln aLLach Lo membrane and forms pore (efflux of lons and nuLrlenLs)
3 klnds alpha beLa eLc (MosL common ls beLa aL anaLomlcal barrlers )
keeps conLrol of growLh of bacLerla
- 9hys|o|og|c (LemperaLure pP Chemlcal MedlaLors)
4 normal bacLerlal flora of lnLesLlne frlendly bacLerla prevenL paLhogen colonlzaLlon
4 Acld pP of sLomach PellobacLer pylorl ulcers ln sLomach (lmmune Lo PCl ln sLomach)
4 Coughlng sneezlng vomlLlng dlarrhoea
- Inf|ammatory
4 vasodllaLlon (vasoacLlve LargeL smooLh muscle ln caplllary Lo relax )
4 lncrease ln caplllary permeablllLy (1lghL [uncLlons relax lncrease permeablllLy)
4 lnflux of phagocyLes
Marg|nat|on leukocyLe sLuck Lo wall and prepare Lo crawl Lhrough
Lxtravasat|on crawllng Lhrough a venule wall
4 Slgns 1lssue redness (eryLhema)relax blood
C|rcu|at|ng Ce||s |n Natura| Immun|ty
- Neutroph||s Larly hagocyLosls LxcellenL kllllng (hlgh resplraLory bursL) of lngesLed mlcrobes
4 Causes collaLeral damage of lnflammaLlon and Llssue damage
4 use preoxldaLlve and free C2 radlcals (hlghly Loxlc)
- Macrophages (Monocytes) LfflclenL hagocyLosls and kllllng of mlcrobes secreLlon of cyLoklnes LhaL sLlmulaLe
lnflammaLlon and lmmune cells
4 Macrophages ls Lhe acLlvaLed from of MonocyLes whlch clrculaLes ln blood
- Nk Ce||s (Natura| k|||er ce||s) nonphagocyLlc lysls of lnfecLed cells acLlvaLlon of macrophages
4 Survelllance of own cells especlally does muLaLed (cancerous)
kecogn|z|ng M|crobes keceptors
keceptor 1ype 1arget
LS LS on Cram ve bacLerla cell wall
nformyl meLhlonyl Commonly found wlLh bacLerla
Mannose ClycosylaLed proLeln wlLh mannoseno
mammal has Lhls
3

- Macrophages
and neuLrophlls
express many
cell surface recepLors LhaL may blnd mlcrobes for subsequenL hagocyLosls







- 1CLL L|ke keceptors (1Lks)
4 Censored Lo cerLaln compoennLs of Lhe bacLerlal cell wall
4 aLLern reorganlzaLlon recepLors of lnnaLe lmmunlLy recognlzes paLhogenassoclaLed molecular
paLLerns" (AMS)
4 Lach 1L8 deLecLs a dlsLlncL subseL of paLhogens lncludlng vlruses bacLerla fungl and proLozoa
4 1246 acL as dlmmers
4 1CLL Cene frulL fly uorsal axel
developmenLal proLeln reslsLs fungal lnfecLlon





9rocess of 9hagocytos|s
Cpson|ns macromolecules LhaL coaL mlcrobes Lo greaLly lncrease lLs recognlLlon by surface recepLors on phagocyLes
1 Attachment medlaLe complemenL cascade
a 8ecepLor recognlse ve charges
b Speclflc opsonln (anLlbody) coaLs mlcrobe Lo lnLenslfy hagocyLosls
l Lall plece of (lgC lc) blnds Lo recepLor
c nonspeclflc opsonln
l C38 (vlLal) complemenL componenL proLeln enhances for rapld dlgesLlon
2 Ingest|on hagocyLosls
a Some bacLerla are encapsulaLed prevenLs macrophage from aLLachlng Lo lL
l S hneumonlae olysaccharlde capsule
ll S yrogenes M Surface proLelns (can cause SLrep LhroaL flesh eaLlng dlsease)
3 Ius|on wlLh lysosomes for mlcrobe degradaLlon
4 |gest|on
3 ke|ease
M|crob|c|da| mechan|sms of Neutroph||s and Macrophages
- Neutroph||s shorLllved afLer hagocyLosls dead neuLrophlls form pus"
- Macrophages Long llved and conLlnue Lo form lysosomes afLer hagocyLosls
1 Lysosomal Lnzymes ln phagolysosomes klll mlcrobes
2 hagocyLe oxldase (membrane proLeln) converLs C2 8Cl (8eacLlve Cxygen lnLermedlaLes)
3 lnCS (lnduclble nlLrlc Cxlde synLhase converLs arglnlne Lo nC (very Loxlc Lo mlcroorganlsm)
C|rcu|at|ng 9rote|ns of Innate Immun|ty
Scavenger 8ecognlze vely charged molecules from
bacLerla (cleanlng up properLy)
keceptor 1ype 1arget
1L81 and 2 8acLerla paraslLes
1L82 and 6 Cram +ve bacLerla and fungl
1L84 Cram ve bacLerla (LS recepLors)
1L83 llagellaLed bacLerla
1L83
1L87 and 8
1L89
ds8nA vlrus
ss8nA vlrus
8acLerla unA dlnucleoLldes vlrus
4

- Comp|ement kllllng of mlcrobes opsonlzaLlon of mlcrobes acLlvaLlon of leukocyLes (lymphocyLes macrophage)
- Mannose8|nd|ng Lect|n (Co||ect|n)
4 8eacL Lo cerLaln sugars on mlcroorganlsms (mosL prevalenL)
4 nonspeclflc opsonlsaLlon of mlcrobes (CoaL Lo enhance hagocyLosls)
4 AcLlvaLlon of complemenL (vla LecLln aLhway)
- Ckeact|ve 9rote|ns (9entrax|n)
4 Ca2+ blndlng domaln on polysaccharlde
4 CpsonlzaLlon of mlcrobes acLlvaLlon of complemenL
4 Measure blood for CreacLlve proLeln b/c lL's marker Lo Lell lf LreaLmenL ls worklng (low levels
LreaLmenL worklng)
- Coagu|at|on Iactors CloLLlng (walllng off lnfecLed Llssue)
Adapt|ve |mmun|ty
AnLlgen dependenL and speclflc
lag Llme response
Memory
L|ements of Adapt|ve Immun|ty
lmmunoglobullns (anLlbodles) Pumoral medlaLed response
8 and 1 LymphocyLes Cell medlaLed response
4 8 cells bone marrow derlved lymphocyLe dlfferenLlaLes lnLo plasma cells and Lhen release anLlbodles
4 1 Cells Lhymus derlved lymphocyLes
Pelper 1 Cells helps 8 cells make anLlbodles
klller 1 Cells recognlze and klll vlral lnfecLed cells
Ieatures of Adapt|ve Immune response
Spec|f|c|ty Lnsures LhaL dlsLlnc mlcrobes ellclLy speclflc lmmune responses permlLs lL Lo dlsLlngulsh subLle
dlfference among forelgn anLlgens
Memory leads Lo enhanced responses Lo repeaLed exposures Lo Lhe same mlcrobe (confer llfelong lmmunlLy)
|vers|ty Lnables LhaL Lhe lmmune sysLem respond Lo a very large varleLy of mlcrobes and molecules
Spec|a||zat|on CeneraLes responses LhaL are opLlmal for defence agalnsL dlfferenL Lypes of mlcrobes
Se|f||m|tat|on Allow mlcrobeacLlvaLed lmmune responses Lo reLurn Lo lLs basal sLaLe (homeosLasls)
|scr|m|nat|on (se|f vs nonse|f) revenLs reacLlve ln[ury Lo hosL's self Llssue durlng responses Lo mlcrobes
3

UNI1 3 Ce||s and Crgans
Crgans and 1|ssue of Immune System
ket|cu|oLndothe||a| System (kLS) ] Mononuc|ear 9hagocyte System
neLwork of phagocyLes ln Llssues
Macrophages resldenL Llssue macrophages
8eLlcular llbres keep cells of Llssue LogeLher
1lssue cells
8raln mlcrogllal cells Lung alveolar macrophages Splenlc macrophages
Lymph node resldenL and
reclrculaLlng macrophages
Synovlal A cells Llver kupffer cells
kldney mesanglal cells 8lood monocyLes recursors ln bone marrow

9r|mary Lympho|d Crgans
8one Marrow Where all blood cells and phagocyLes orlglnaLe 8 cells maLure here
1hymus Where 1 cells compleLe Lhelr developmenL afLer produced ln bone marrow
Secondary Lympho|d 1|ssues
Lymph nodes Lrap local Llssue anLlgen
4 ConLalns a capsule
4 A corLex fllled wlLh 8 lymphocyLes + folllcular uendrlLlc cells
rlmary folllcle 8 cells Cermlnal cenLers ln Lhe folllcle conLalns prollferaLlng 8 cells
4 aracorLex 1 and lnLerdlglLaLlng uendrlLlc cells
4 Medullary cords 8 1 and plasma cells
4 Medulla plasma cells
4 1he Plgh endoLhellal venule (PLv) ls where blood enLers lnLo Lhe lymph Llssues Lo be fllLered
uendrlLlc Cells process and presenL anLlgens Lo 1 Cells
4 Langerhan Cells found ln epldermls mucosal membranes
4 lnLersLlLlal uendrlLlc cells populaLe mosL organs
4 lnLerdlglLaLlng cells found ln 1cell areas of secondary lymphold Llssues and Lhymlc medulla
4 ClrculaLlng uendrlLlc cells found ln blood lymph (velled cells)
CapLure anLlgen ln skln cuL and Lransfer Lo lymph Lo presenL
4 lolllcular uendrlLlc cell nC1 anLlgen presenLlng cell (AC)
LocaLed ln 8 cell rlch folllcles
express hlgh levels of membrane recepLors for anLlbody and complemenL
8lnd clrculaLlng AbAg complexes Lo faclllLaLe 8 cell acLlvaLlon ln Lymph nodes
Spleen SlLuaLed hlgh ln lefL abdomlnal cavlLy Lo fllLer blood and Lrap bloodborne mlcrobes (anLlgen)
4 CapsulaLed
ArLery + velns surrounded by per|arter|a| |ymphat|c sheath (9ALS) rlch ln 1 cells
Marglnal zone nexL layer and rlch ln 8 uendrlLlc cells
4 8ed ulp removes old and defecLlve red cells
4 WhlLe ulp lymphocyLe acLlvaLlon
4 rlmary folllcle 8 Cells
4 SplenecLomy leads Lo lncrease bacLerlal lnfecLlon Causes
Cancer ln spleen 88C are belng desLroyed rupLure from conLacL sporL or car accldenL
4 eyer's paLches ln lleum of ShorL lnLesLlne sample lnvadlng mlcrobes aL Mcells
6

Mucosal Lymphold 1lssue
4 roLecLs aL mucosal (MAL1) eplLhellal surfaces
4 CAL1 ulgesLlve 8AL1 8esplraLory and urogenlLal LracL
4 bulk of lymphold Llssues (400m
2
) very large Llssue
4 Some organlzaLlon (folllcles)
4 MAL1 ls blased Lowards lgA producLlon and has own speclallzed lymphocyLes (mucosal homlng
recepLors) lymphocyLes are able Lo mlgraLe ouL and populaLe oLher areas
Lymphat|c System
MosL of Lhe Llssues organs connecLlng vessels and clrculaLlng cells
1lssues are called lymphaLlc Llssues
lnLerconnecLlng vessels are called Lhe lymphaLlcs
Lhe lymphaLlcs LransporL Llssue fluld known as lymph whlch orlglnaLed as Lhe llquld plasma of Lhe blood
Ce||s of the Immune System 8|ood Ce||s
ked 8|ood Ce|| (k8C) 3 x 10
6
cells/mm
3

9|ate|ets Small cyLoplasmlc sLrucLures derlved from megakaryocyLes conLaln cloLLlng facLors 23 x 10
3
cells/mm
3

Leukocytes (W8C) 73 x 10
3
cells/mm
3

neuLrophlls 3070 generaLes fasL shorL halfllfe hagocyLlc granulocyLes lnflammaLory medlaLors
LymphocyLes (8 and 1 Cells and nk) 2040 nk cells produce cyLoklnes LhaL shares Lhe same lnLerferon gamma
wlLh 1 cells nk cells also recognlzes vlruslnfecLed and Lumor cells
MonocyLes 16 crlLlcal for lnnaLe and speclflc lmmunlLy phagocyLlc
4 ulfferenLlaLe lnLo macrophage and uendrlLlc cells
4 roduce cyLoklnes proLelns for communlcaLlng and acLlvaLlon
4 roduce ComplemenLs proLelns for lnflammaLlon
Loslnophlls 13 hagocyLlc granulocyLes lnflammaLory medlaLor
8asophlls 1 (responslble for allerglc reacLlons) granulocyLes lnflammaLory medlaLors
Mast Ce||s
Ce||s of Innate Immune kesponse All granulocyLes MonocyLes nk cells and MasL Cells
Ce||s of Spec|f|c Immune system
8 Lymphocytes agalnsL exLracellular phases of bacLerlal and vlral lnfecLlons and Loxlns
uevelops ln Lhe bone marrow
roduce anLlbody afLer anLlgen sLlmulaLlon (dlfferenLlaLlon) lnLo plasma cells
have speclallzed anLlgen recepLors (eg surface anLlbody)
8ecognlzes and blnd Lo anLlgen
CLher characLerlsLlc surface markers
4 ComplemenL recepLor C81 and 2
4 822 frequenLly used as a marker for b cells
4 87 lmporLanL regulaLory molecule
4 Cu40 crlLlcal for survlval of anLlgensLlmulaLed 8 cells
8 cells and AC (Macrophage uendrlLlc cell) presenLs anLlgen Lo Pelper L cell for furLher lmmune response
7

1 Lymphocytes
MaLure ln Lhymus
speclallzed surface recepLor for recognlzlng pepLlde anLlgens bound Lo MPC molecules 9class 1 and class 2)
CLher characLerlsLlcs markers (eg Cu8 Cu4)
Pelper 1 cell (Cu4)
4 Pelper 1cell 1 become effecLor kllllng cell produce cyLoklne LhaL can acLlvaLe klller 1 and nk cells
4 Pelper 1cell 2 help sLlmulaLe 8 cells' producLlon of anLlbodles
8egulaLory 1cell Suppresses Lhe LlymphocyLes once lnfecLlon ls over
klller 1cell (Cu8) kllls vlral fungal and paraslLlc cells vla anLlgen bound MPC class 1
4 AgalnsL lnLracellular vlral and bacLerlal lnfecLlons Lumor cells LransplanLed organs / Llssues
8

UNI1 4 Ant|gens Immunogen|c|ty
Immunogen A molecule LhaL has Lhe ablllLy Lo evoke a speclflc lmmune response and can reacL wlLh Lhe resulLanL
speclflc anLlbody
Ant|gen A molecule LhaL can reacL wlLh a speclflc performed anLlbody or speclflc recepLors on 8 cells (Moy ot moy oot
be lmmoooqeolc some bos to bloJ to o ptotelo to evoke lmmooetespoose)
Iactors that Inf|uences Immunogen|c|ty
Contr|but|on of Immunogens
lorelgnness CreaLer Lhe hylogeneLlc dlsLance b/w Lwo specles Lhe greaLer molecular sLrucLural dlsparlLy ln Lhe
molecules beLween Lhem
4 oor lmmunogen lsogenlc or auLologous (self) Syngenelc (same le ldenLlcal Lwlns) Allogenelc
(dlfferenL lndlvldual of Lhe same specles) xenogenelc (dlfferenL specles) 8esL lmmunogen
Molecular Slze generally molecules wlLh molecular mass 10000ua are good lmmunogens
4 oor lmmunogens lnsulln PlsLones CxyLocln
4 Cood lmmunogens ovalbumln rlbonuclease lmmunoglobulln
Molecular ComplexlLy lnLernal molecular complexlLy ls essenLlal for good lmmunogens
4 SynLheLlc homopolymers of slngle AA or sugars are noL lmmunogenlc (sLrucLurally unsLable)
4 AddlLlon of aromaLlc AA llke Lyroslne phenylalanlne enhances Lhe lmmunogenelclLy
CelaLln poor lmmunogen
CelaLlnLyroslne excellenL lmmunogen
uegradablllLy AnLlgens LhaL are easlly phagocyLosed are generally more lmmunogenlc
4 lor more anLlgens Lhe developmenL of an lmmune response requlres LhaL Lhe anLlgen be phagocyLosed
processed and presenLed Lo helper 1 cells by an AC
4 SynLheLlc polyers of uamlno aclds (naLurally occurrlng are LAA) can noL be processed by enzymes and
Lhus are poor lmmunogens
hyslcal form(qeoetolly) arLlculaLe (aggregaLes of proLelns) anLlgens are more lmmunogenlc Lhan soluble ones
4 denaLured anLlgens are more lmmunogenlc Lhan LhaL naLlve form
Contr|but|on of 8|o|og|ca| System
CeneLlflcally llL Lo produce an lmmune response
4 MounLlng lmmune response musL be able Lo make MPC class 1 or 2 recepLors
uefecL won'L mouLn good response
lmmunodeflclency can'L make lmmunoglobullns
CpLlmal uose of lmmunogen wlLh repeaLed ln[ecLlons over a perlod of weeks ls requlred
4 1oo llLLle Lolerance or noLhlng
4 1oo Much damper
8ouLe of anLlgen admlnlsLraLlon sLrongly lnfluences whlch lmmune organ and cell populaLlons wlll be lnvolved
ln lmmune response
4 SubcuLaneous or lnLramuscular good response ln dralnlng lymph nodes
4 lnLravenous general (sysLemlc) response especlally ln Lhe spleen
4 lnLranasal oral mucosal response
Cral 1olerance makes lL dlfflculL Lo mounL response


9

Ad[uvanLs SubsLances when mlxed wlLh anLlgens and ln[ecLed wlLh lL enhances Lhe lmmunogenlclLy of LhaL
anLlgen and noL Lhe ad[uvanL
4 ln[ecLlon of anLlgen body waLer dlluLe anLlgen llver dlgesLs lL
wanL lnflammaLory response waxy subsLance Lo hold anLlgen
4 kllled organlsm can enhance anLlgenlclLy
4 used ln vacclnes (eg alumlnum poLasslum sulfaLe)
8efore used kllled 8ordeLella erLussls whooplng cough
4 lreund's waLerlnoll ad[uvanL emulslon agenL Lo keep Lhe soluLlon LogeLher
CompleLe (conLalns kllled mycobacLerlum)
lncompleLe (no kllled bacLerla)
4 Several mechanlsms
keep lmmunogen locallzed
Lnhance lnflammaLlon waxy maLerlal dropped on waLer allows lmmune cell Lo come Lo
ln[ecLlon
Concepts of Lp|topes
Lp|tope ls Lhe parL of Lhe anLlgen LhaL blnds wlLh Lhe anLlbody blndlng slLe or anLlgenspeclflc membrane recepLors on
lymphocyLes
9ropert|es of 8 Ce|| and Ant|body Lp|topes
LplLopes are Lhe lmmunologlcally acLlve reglon of an anLlgen LhaL blnd Lo anLlgenspeclflc membrane recepLors
on lymphocyLes or Lo secreLed anLlbodles
Comparlson of AnLlgen recognlLlon by 1 cells and 8 cells
Character|st|cs 8 Ce||s 1 Ce||s
Interact|on w|th ant|gen lnvolves blnary complex of
membrane lg and Ag
lnvolves Lernary complex of 1
cell recepLor Ag and MPC
molecule
8|nd|ng of so|ub|e ant|gen ?es no
Invo|vement of MnC none requlred 8equlred Lo dlsplay processed
anLlgen
Chem|ca| nature of ant|gen roLeln polysaccharlde llpld MosL proLelns buL some llplds
and glycollplds presenLed on
MPCllke molecule (le Cu 1)
Lp|tope propert|es Accesslble hydrophlllc moblle
pepLldes conLalnlng sequenLlal or
nonsequenLlal amlno aclds
lnLernal llnear pepLldes
produced by processlng of
anLlgen and bound Lo MPC
MulLlvalenL anLlgen wlLh dlfferenL eplLopes resulL ln polyclonal lmmune response (more Lhan 1 anLlbody Lype
requlre Lo blnd Lo anLlgen)
MulLlvalenL anLlgen wlLh repeaLlng epLlopes resulL ln monoclonal lmmune response (1 klnd of anLlbody ls
needed)
AnLlbody blnds Lo an anLlgenlc eplLope by weak noncovalenL lnLeracLlons whlch operaLe only over shorL
dlsLances (le Pbondlng London forces lonlc and hydrophoblc)
AnLlbody's blndlng slLe and Lhe eplLope musL have complemenLary shapes (lock and key)
lnLeracLlng surfaces can be flaL or undulaLlng
8 cell epLlopes on naLlve proLelns are generally composed of Pydrophlllc amlno aclds (eg gluLamlc acld and
Lyroslne) on Lhe proLeln surface LhaL are Lopographlcally accesslble Lo membrane bound or secreLed anLlbody
10

8cell eplLopes can conLaln sequenLlal (llnear) or nonsequenLlal (dlsconLlnuous) amlno aclds locaLed ln flexlble
reglons of Lhe anLlgen
Complex proLelns conLaln mulLlple overlapplng 8 cell eplLopes some of whlch are lmmunodomlnanL (le lnduce
a more pronounced lmmune response)
Concept of naptens
napten small organlc molecule LhaL ls anLlgenlc buL noL lmmunogenlc unless lL ls aLLached Lo a large proLeln called
carrler
Lxample ulnlLrophenol (un) and 8SA (8ovlne serum albumln proLeln carrler)
Interact|ons w|th Ant|bod|es formed
PapLen (un) none
roLeln Carrler (8SA) AnLl8SA
PapLencarrler con[ugaLe (unA + 8SA) AnLlun
AnLl8SA
AnLlunA/8SA
CLher Lxamples of PapLens
4 urug allergles penlclllln can reacL wlLh proLelns Lo form a penlcllloylproLeln derlvaLlve
4 Amlnobenzene and derlvaLles
Concept of Spec|f|cty AnLlbodles can dlscrlmlnaLe beLween Lwo anLlgens lle show speclflclLy by Lhelr greaLer afflnlLy
for one raLher Lhan Lhe oLher
Spec|a| character|st|cs of ant|body spec|f|c|ty requlremenL for accesslblllLy of anLlgenlc deLermlnanLs (epLlopes) and
sLerochemlcal conslderaLlon
LplLopes of sLrong acldlc groups conLrlbuLed Lo anLlbody speclflclLy
LplLopes of nonlonlc radlcal groups conLrlbuLes Lo anLlbody crossreacLlons lmmune response Lo oLher
eplLopes oLher Lhan lLself (le blood A8C anLlgen)
LplLopes of glycoslde groups conLrlbuLes Lo exLreme anLlbody speclflclLy
Concept of neteroc||t|c ant|bod|es
1hese anLlbodles have hlgher afflnlLy for crossreacLlng eplLopes Lhan for Lhelr homologous eplLope
4 AnLln anLlbodles have hlgher afflnlLy for cross reacLlng nl eplLope Lhan for Lhelr homologous n
eplLope
4 lmmunlzlng epLlope ls 3nlLro4hydroxyphenylaceLyl (n
4 CrossreacLlng eplLope ls 3nlLro3lodo4hydroxyphenylaceLyl (nl)
11

UNI1 #S Immunog|obu||ns Structure and Iunct|on
8ackground and ef|n|t|ons
Immunog|obu||ns funcLlons as anLlbodles AnLlbodles effecLor molecules of humoral lmmunlLy LxlsLs as
SecreLed anLlbodles by plasma cells (dlfferenLlaLed 8 cells)
AnLlgenblndlng recepLors Cne 8 cell membranes
9|asma Cell free fluld from blood whlch conLalns cloLLlng facLors
Serum 8esldual fluld when blood ro plasma forms a cloL ConLalns many anLlbodles
Ant|serum Serum conLalnlng anLlbodles LhaL blnd Lo a parLlcular anLlgen (only 1 Lype of anLlbody)
Sero|ogy 1he sLudy of anLlbodles and Lhelr reacLlons wlLh anLlgens
9o|yc|ona| Ant|bod|es Serum conLalns mlxLures of dlfferenL anLlbodles produced by many clones of 8 lymphocyLes
ulfferenL anLlbodles produced Lo flghL agalnsL Lhe same anLlgen LhaL has dlfferenL epLlopes
8reakthrough |n Ant|body Structure
Mu|t|p|e Mye|oma Monoclonal Lumor of anLlbodyproduclng plasma cells (8 cells) maklng ldenLlcal anLlbody molecules
Myeloma cells also secreLe eoceIooes ptotelosexcesslve amounLs of llghL chalns found ln urlne
nybr|doma anLlbody produclng cell llne
1echnlque of produclng unllmlLed quanLlLles of ldenLlcal anLlbody molecules
luslng a muLanL myeloma cell llne wlLh spleen cells from a mouse lmmunlzed wlLh anLlgen
Pow does lL work?
4 llrsL lsolaLe 8 cells from lmmunlzed mouse cells
4 luse MuLanL myeloma cell ln PA1 medlum
MuLanL myeloma cells have muLaLlon ln PC81 and 1k and can noL grow ln PA1 b/c Lhe
AmlnopLerln blocks proLeln synLhesls PC81 and 1k can flx Lhls buL slnce Lhelr muLanL Lhey dle
lf grown alone
4 When fused PC81 and 1k funcLlons reLurn and Lhe fused cells produce unllmlLed supply of Lhe same Ab
enera| Iunct|ons and 9ropert|es of Immunog|obu||ns
AnLlgen 8lndlng prlmary funcLlon of anLlbodles resulLs ln proLecLlon of Lhe hosL
4 valency Lhe valency of an anLlbody refers Lo Lhe number of anLlgenlc deLermlnanLs LhaL an lndlvldual
anLlbody molecule can blnd
LffecLor luncLlons Ab cannoL klll or ellmlnaLe paLhogens slmply by blndlng Ag slgnlflcanL blologlcal effecLs are
a consequences of secondary effecLor funcLlons"
4 AcLlvaLlon of complemenL sysLem lysls of cells clearance of lmmune complexes (AbAg) ln blood
4 neuLrallzaLlon of paLhogens by lnLerferlng wlLh enLry lnLo hosL cells
4 8lndlng Lo varlous cell Lypes vla lc recepLors
hagocyLlc cells opsonlzaLlon
MasL cells and basophlls allergy (lgL)
nk cells AuCCC anLlbodydependenL cellmedlaLed cyLoLoxlclLy
12

8as|c Structure of Immunog|obu||ns
lg can dlffer sLrucLurally 8u1 Lhey have Lhe same baslc unlL PeLerodlmer and composed of 4 chalns
ComposlLlon of lg SLrucLure
4 Peavy Chaln (P) and LlghL Chaln (L) Lwo ldenLlcal llghL and heavy chalns
4 ulsulflde bonds
lnLerchaln P and L held LogeLher P and P held LogeLher
lnLrachaln wlLhln each of Lhe polypepLlde chalns
4 varlable (v) and ConsLanL reglons
LlghL chaln vL (110aa) and CL (110aa)
Peavy Chaln vP (110aa) and CP (330440aa)
4 Plnge 8eglon reglon whlch Lhe arms of Lhe anLlbody molecule form a ?"
1hls allows for some flexlblllLy ln Lhe molecule aL Lhls polnL (excepLlon lgM and lgL)
4 uomalns globular reglons each conLaln an lnLrachaln dlsulflde bond
lmmunoglobulln domalns are folded lnLo a characLerlsLlc sLrucLure lmmunoglobulln lold"
LlghL Chaln vl and Cl
Peavy Chaln vP CP1 CP3 (or CP4)
4 Cllgosaccharldes carbohydraLes are aLLached (nllnked) Lo Lhe CP2 domaln ln mosL lmmunoglobullns
1he lmmunoglobulln Super lamlly
4 Large famlly of cell membrane proLelns
4 osses one or more reglons homologous Lo Lhe lmmunoglobulln fold
4 uomalns arranged ln pleaLed sheeLs of anLlparallel beLa sLrands
4 Lxamples Lcell recepLors Cu4 Cu8 Cu3 MPC class l and ll
Pypervarlable 8eglongs (Pv) complemenLarlLy deLermlnlng reglons (Cu8) Llp of anLlbody blndlng slLes
lmmunoglobulln lragmenLs lormed by LreaLmenL of anLlbodles wlLh proLeases
4 roLeases proLeolyLlc enzymes LhaL cleave pepLldes aL speclflc sequences
4 used for dlssecLlng Lhe sLrucLure and funcLlon of anLlbody fragmenLs
4 roLelyLlc cleavage by AAln CreaLes 2 lab lragmenL anLlgen blndlng and 1 lc lragmenL
crysLalllzaLlon
4 roLelyLlc cleavage by epsln creaLes 1 (lab)2 lc geLs degraded
4 lmporLance lc (blnd effecLor cell) and (lab)2 (blnd anLlgen) can be used as Lherapy or ln research
numan Immunog|obu||n C|asses and Subc|asses
AnLlbody class deLermlne by Lhe heavy chaln
AnLlbody subclass ls named ln order of abundances ln serum (1 mosL abundanL) and only lgC and lgA have
subclasses
ln humans 60 of llghL chalns are and 40 are
ln mlce 93 of llghL chalns are and 3 are
Classes of lmmunoglobullns dlffer ln slze charge amlno acld composlLlon and carbohydraLe conLenL
Ig 9ropert|es 9 mg/ml ln serum (80)
uomlnanL lg ln lmmune response
Ma[or anLlmlcroblal acLlvlLy ln plasma and
exLravascular flulds
acLlvaLes complemenL when bound Lo anLlgen
blnds Lo lc recepLors (lcy8) on varleLy of cells
Crosses lacenLa
4 lgC subclasses roperLles ulffer ln slze of hlnge reglon # of SS bonds and amlno aclds
4 lgC1 lgC3 lgC4 cross placenLa (nC1 lgC 2)
13

4 lgC 3 mosL effecLlve complemenL acLlvaLor (lgC3 lgC1 lgC2 lgC4)
4 lgC4 does noL acLlvaLe complemenL
4 lgC 1 lgC3 blnd wlLh hlgh afflnlLy Lo lc recepLors on hagocyLlc cells medlaLlng opsonlzaLlon
IgA 9ropert|es 3 mg/ml (1013)
ulmer ln secreLlons (Lears sallva breasL mllk bronchlal mucous Cl LracL) 8u1 a monomer ln serum
2 addlLlonal proLelns Lo hold Lhe 2 monomers LogeLher
4 ! chaln produced by Lhe 8 cells
4 SecreLory componenL derlved from Lhe eplLhellal cells from whlch lgA ls secreLed
Masks proLease cleavage slLes ln hlnge reglon so Lherefore can lasL longer
lormaLlon lgA ulmer blnds vla ! chaln Lo Lhe oly lg 8ecepLor on eplLhella cell LogeLher recepLor and lgA ls
phagocyLosed lnLo cell goes Lhrough cell and secreLed ouL wlLh oLher flulds
313g of lgA ln mucous dally
Ma[or anLlmlcroblal acLlvlLy ln secreLlons
AcLs malnly as a neuLrallzlng anLlbody b/c lc complex blnded Lo each oLher
4 revenLs aLLachmenL of paLhogens Lo mucosal eplLhellal cells
uoesn'L acLlvaLe classlcal complmenL paLhway
uoesn'L cross placenLa
lgA lsoLypes
4 lgA1 found ln serum [ 101 raLlo (lgA1lgA2)
redomlnanLly monomerlc (consLlLuLe 1013 serum lgs)
olymerlc forms are someLlmes seen
Pave longer hlnge LhaL makes lL less proLecLed Lhus noL ln secreLlon
4 lgA2 found malnly aL mucosal slLes [ 32 raLlo (lgA2 lgA1)
lgA prlnclpal anLlbody class ln secreLlon
Pave nglycans whlch proLecLs ls more
IgM 9ropert|es 13mg/ml (310)
enLamer 3 ?shaped molecules [olned by dlsulphlde bonds and a ! Chaln
8 Cell curface anLlgen recepLor (Slngle ?) (WlLh lgu)
llrsL Lo appear ln phylogeny onLogeny and ln Lhe normal lmmune response
Largely conflned Lo blood sLream
naLural anLlbodles (eg anLlblood group A 8)
Cood aggluLlnaLor (crossllnk Ag well)
LxcellenL complemenL acLlvaLor afLer Ag blndlng
Lack hlnge reglon 8uL lack of hlnge does noL equal lack of flexlblllLy b/c have C2P llnk domaln
Ig 9ropert|es 003mg/ml (02)
Largely conflned Lo membrane of maLure 8 cell LogeLher wlLh membrane lgM
noL expressed on lmmaLure 8Cells
no known blologlcal effecLs


14

IgL 9ropert|es 00003mg/ml (0001)
SllghLly larger more rlgld Lhan lgC
no hlnge
8lnds Lo lc recepLors (lce8) on masL cells and basophlls
4 leads Lo degranulaLlon (release of hlsLamlne) allerglc manlfesLaLlon
Classlcal anLlbody anaphylaxls (1ype l allergles) and cerLaln paraslLlc lnfecLlons
normally very low levels ln Lhe serum
Ant|gen|c eterm|nants of Ig Mo|ecu|es
AnLlbodles have anLlgenlc deLermlnanLs oLher anLlbodles can blnd oL Lhem
Isotype refers Lo dlfferences ln P and L chalns LhaL dlsLlngulsh each class and subclass (eg lg classes C M A u L )
Mouse A's lgC1 vs Mouse 8's lgM
A||otypes dlfferences ln Lhe P and L chalns of same lsoLype buL recognlze Lhe same anLlgen
Mouse A's lgC1 vs Mouse 8's lgC1 (SLraln A vs SLraln 8)
1he lsoLype remalns Lhe SAML Lhe recognlzed anLlgen remalns Lhe SAML
Id|otypes anLlgenlc deLermlnanL (eplLope) ln Lhe anLlgenblndlng slLe of newlyformed anLlbody
Mouse lgC1 agalnsL anLlgen A vs Mouse lgC1 agalnsL anLlgen 8
1he lsoLype remalns Lhe SAML Lhe recognlzed anLlgen ls dlfferenL
13

UNI1 #6 Ant|body 8|osynthes|s
eve|opment of a genet|c Mode|
ene kearrangement recomblnaLlon of gene segmenLs Lo produce a funcLlonal varlable reglon sequence
nonlymphold cells gene encodlng lgs are lA8 aparL
8 Cells genes encodlng lgs are CLCSL LogeLher 1hls ls a resulL of Somat|c recomb|nat|on
Cene 8earrangemenL accounLs for
4 CapaclLy Lo make up Lo 10
11
anLlbodles
4 varlable and consLanL domalns ln same chaln
4 ulfferenL classes of anLlbodles wlLh some anLlgenlc speclflclLy
ene Segments |n L|ght neavy Cha|ns
L|ght Cha|n
varlable uomaln varlable + !olnlng reglon
ConsLanL uomaln ConsLanL reglon
neavy Cha|n
varlable uomaln varlable + ulverslLy + !olnlng reglons
ConsLanL uomalns consLanL reglon
Crgan|zat|on of human |mmunog|obu||n L|ght Cha|n germ ||ne ene segments
Lncoded by L (leader) v (varlable) ! 9!olnlng) and C (ConsLanL) gene segmenLs
LlghL Chaln Cn Chromosome 22 ! segmenLs parLners wlLh lLs ad[olnlng C segmenL
4 30 varlable reglons 4 [olnlng reglons
LlghL Chaln Cn Chromosome 2 ! segmenL can [oln wlLh Lhe same C segmenL (only 1 C)
4 40 varlable reglons 3 [olnlng reglons
Crgan|zat|on of human |mmunog|obu||n neavy Cha|n germ||ne gene segments
Lncoded by L (leader) v (varlable) ! (!olnlng) u (ulverslLy) and C (ConsLanL) gene segmenLs
1here are 8 consLanL segmenLs each one for a speclflc class / subclass of Lhe 8 posslble Lypes of anLlbodles
40 varlable reglons 23 dlverslLy reglons 6 [olnlng reglons
Cverv|ew of ene kearrangement base of an IgM Ant|body
VI rearrangement |n neavy Cha|n NA
1 1
sL
8earrangemenL ls LhaL a u and ! geLs [olned LogeLher LveryLhlng ln beLween geLs exclsed
2 2
nd
rearrangemenL ls LhaL v geLs added Lo u! LveryLhlng ln beLween geLs exclsed
1 ooJ 2 ote ecomblootloo ctlvotloq eoe ozyme tespooslble fot tbe exclsloo ooJ teottooqemeot
3 vu! CCCCCCCC goes Lhrough LranscrlpLlon
4 8nA processlng spllclng ouL of Lxcess 8nA"
3 AddlLlon of a olyA Lalls m8nA exlLs nucleus Lo blnd rlbosomes ln rL8
6 1ranslaLlon of m8nA L sequence pulls growlng polypepLlde lnLo lumen of L8 ls Lhen cleaved off
7 vu!C ready Lo go Lo c ell surface
16

VI rearrangement |n L|ght Cha|n NA
1 Cnly 1 rearrangemenL v [olned Lo ! (vla 8AC 1 and 8AC 2)
2 LranscrlpLlon
3 8nA processlng and spllclng (removal of excess C domalns and addlLlon of polyA Lall)
4 Moves ouL of nucleus and blnd Lo rlbosomes ln rL8
3 1ranslaLlon L sequence pulls growlng polypepLlde lnLo lumen of L8 ls Lhen cleaved off
6 v!C ready for secreLlon
Cne 1urn ] twoturn ku|e for recomb|nat|on S|gna| Sequences (kSS)
1 Lurn 1 Lurn of Lhe unA hellx
2 Lurn 2 Lurn of Lhe unA hellx
v ! and u reglons have a 8SS Lag of elLher 1 Lurn or 2 Lurn
Cnly 1 Lurn 8SS can blnd Lo 2 Lurn 8SS 1hls prevenLs lmproper [olnlng
9roduct|ve and nonproduct|ve kearrangements
8AC cleavage can be anywhere wlLhln Lhe 8SS Lurn 1hls glves varlaLlon by funcLlonal flexlblllLy wlLhln anLlgen
blndlng slLe
Adds Lo anLlbody dlverslLy
1he resulLlng proLeln can be producLlve (no SLop codon) or nonproducLlve (have sLop codon)
key Concept one 8 Cell makes anLlbodles LhaL recognlze one eplLope lC8 LllL
vu! and v! rearrangemenLs are done CnCL ln Lhe 8 cell llfe span
Concept of A||e||c Lxc|us|on
Lnsures LhaL funcLlonal 8 cells never conLaln more Lhan one vu! amnd v! unlL
Cne 8 cell wlLh one anLlgenlc speclflclLy
neavy Cha|n 9roduct|on lrom ro8 cell Lo re8 cell
Peavy chaln vu! rearrangemenL ln flrsL allele roducLlve 8nA proLeln pre8 recepLor expresslon
lnhlblLlon of furLher heavy chaln gene rearrangemenL llghL chaln producLlon
lf unproducLlve vu! rearrangemenL ln flrsL allele heavy chaln vu! rearrangemenL ln 2
nd
allele
lf 2
nd
allele arrangemenL ls unproducLlve Cell deaLh
L|ght Cha|n 9roduct|on re8 cell Lo lmmaLure 8 cell
chaln v! rearrangemenL ln 1
sL
allele lf unproducLlve Lhan 2
nd
allele of chaln
lf unproducLlve Lhan vP rearrangemenL ln 1
sL
allele of chaln lf unproducLlve Lhan 2
nd
allele of chaln
lf producLlve afLer any one of Lhe rearrangemenLs roducLlon of Lhelr respecLlve 8nA chaln proLeln and lgm
expresslon (le or ) lnhlblLlon of furLher llghL chaln gene rearrangemenL
lf no rearrangemenL are producLlve cell deaLh
Membrane 8ound vs Secreted Ant|bod|es
P Chaln prlmary LranscrlpLs undergo dlfferenLlal 8nA processlng
4 lmmaLure b cells membrane bound lgM
4 MaLure 8 cells membrane bound lgM and lgu
17

4 lasma Cells secreLed anLlbodles
CeneraLlon of Membrane lgM / lgu
4 new" 8 cells favour polyadenylaLlon slLes 2 4
4 Membrane exon 1 2 lncluded ln flnal m8nA encodlng membrane Chaln
4 CeneraLlon of lgM and lgu occur slmulLaneously
CeneraLlon of secreLed lgM
4 lasma cells vaour polyadenylaLlon slLe 1 (lgM) or 3 (lgu)
4 no Membrane exons lncluded
C|ass Sw|tch|ng
Cccurs upon anLlgenlc sLlmulaLlon of c cell
lurLher gene rearrangemenL vu! can comblne wlLh a C
h

4 ln beLween each C
P
Lhere`s a swlLch slLes 8eglons of unA recognlzed by enzymes LhaL regulaLe class
swlLchlng
4 C
u
does noL have a swlLch slLe
Cccurs aL Lhe level of unA
Class swlLchlng moves forward only lgM can swlLch Lo lgC lgA lgL buL noL lgA Lo lgM
rocess lnvolves unA looplng ouL of Lhe C
P
noL needed by Lhe enzyme acLlvaLlonlnduced CyLldlne ueamlnase
(Alu) 8nA rocesslng AnLlbody
Class swlLchlng can occur more Lhan once as long as ln forward moLlon lgC lgC lgL
enet|c bas|s for Ant|body |vers|ty
1 MulLlple germllne genes conLrolllng v reglons
2 MulLlple v! and vu! comblnaLlon
a Clve rlse Lo dlfferenL Agblndlng speclflclLles
3 varlablllLy ln v! vu or u! [olnlng ([uncLlonal flexlblllLy)
a varlaLlon aL one or Lwo poslLlons ln Lhe Cu83 hypervarlable reglons
4 nucleoLlde addlLlons ln heavy Chaln (ln beLween breaks durlng rearrangemenL of dlfferenL reglons)
a nucleoLlde addlLlon resulLs ln pallndormlc sequence
b nnucleoLlde addlLlon ls a 8andom addlLlon fo nucleoLldes by Lnzyme 1ermlnal ueoxyrlbonucleoLldyl
1ransferase (1d1)
3 SomaLlc MuLaLlons (Alu enzyme)
a SomaLlc muLaLlons (random process) ln lg genes ln 8 cells (v[ and vu! unlLs unA)
b Cccurs only ln germlnal cenLers
c AA subsLlLuLlons
d lrequenLly ln CuS's
e lrequency raLe ls 100000 Llmes hlgher Lhan Lhe sponLaneous raLe ln oLher genes
f AssoclaLed wlLh classswlLchlng
g AfflnlLy maLuraLlon an lncrease ln afflnlLy of Ab for Ag durlng Lhe course of lmmaLure response or ln
subsequenL exposures Lo Ag
6 ComblnaLlonal AssoclaLlon of P and L Chaln
a alrlng of a glven L chaln wlLh adlfferenL P chaln may glve rlse Lo dlfferenL speclflclLles
b 2000 L 1000 P Chalns 2 x10
6
dlfferenL anLlbody speclflclLles by random comblnaLlons

18

UNI1 #7 8Ce|| enerat|on
Cverv|ew of 8 Ce|| L|fe
8 Ce|| precursor rearranges |s |mmunog|obu||n genes" CeneraLlon of C cell recepLors ln Lhe 8one marrow
Immature 8 ce|| bound to se|f ce||surface ant|gen |s removed from the reperto|re negaLlves seklecLlon ln Lhe
bone marrow
Mature 8 ce|| bound to fore|gn ant|gen |s act|vated mlgraLlon of C cells Lo perlpheral lymphold organs and
acLlvaLlon
Act|vated 8 ce|| g|ve r|se to p|asma ce||s and memory ce||s anLlbody secreLlon and mmeor cells ln bone marrow
and lymphold Llssues
8 Ce|| Maturat|on |n the 8one Marrow Ant|gen Independent 9hase
rogenlLor 8 cell have Cu438 surface marker (LeukocyLe Common AnLlgen")
Make abouL 3 mllllon 8 cells a day
Cnly 10 of 8 cell leaves Lhe bone marrow
9rocess Ig kearrangement 8Cell developmenL depends on Lhe producLlve sequenLlal rearrangemenL of P Chaln and L
chaln genes
Lymphold rogenLlor cells blnds Lo vCAM1 (vascular adheslon molecule1) vla Lhelr lnLegrln vLA4 (very laLe
anLlgen4)
4 u! rearrangemenL of Peavy chaln by 8AC 1 and 8AC 2 ro 8 Cell
ro8 Cell 1hls promoLes Lhe blndlng of early pro8 cell surface cklL recepLor (le Lyroslne klnase) Lo sLemcell
facLor (SCl) on Lhe sLromal cell surface AcLlvaLes Lhe klnase and lnduces cell acLlvaLlon vu! rearrangemenL
of lgM heavy chaln
4 ConLalns lg and lg a heLerodlmer complex requlred for slgnalllng when Ag blnds Lo Ab on surface
of maLure 8 cell
4 vu! rearrangemenL of heavy chaln vla 8AS (8 Cell speclflc acLlvaLor proLeln) re8 Cell
LaLe ro8 Cell 8ecepLors for growLh facLor lL7 requlre boLh SCl/klL blndlng and soluble lL7 for Lhelr growLh
and maLuraLlon
re8 Cell Lose Lhelr dependence on SCl and cease Lo express cklL AbsoluLely requlre lL7 Lo drlve prollferaLlon
and survlval of pre8 cell
4 have SurrogaLe LlghL Chaln vpre8 (varlablellke 8) and 3 (ConsLanL llke chaln)
helps move heavy chaln Lo Lhe surface
1hls chaln complex conLalnlng Lhe surrogaLe llghL chaln cross llnks wlLh sLromal cell logand
(proLelns express by sLromal cell)
4 SLops furLher heavy chaln gene rearrangemenL
4 lnduces llghL chaln rearrangemenL (v!) lmmaLure 8 cell
lmmaLure 8 Cell no longer express surrofaLe llghL chalns buL lnsLead express or llghL chalns LogeLher wlLh
Lhe heavy chaln formaLlon of lgM surface recepLor
4 Crossllnklng by anLlgen
Cell ueaLh lf recognlze self anLlgen
AcLlvaLlon slgnal maLuraLlon sLep for lgu
Negat|ve se|ect|on (C|ona| e|et|on)
MPC class l self proLelns P2
d/k
are on Lhe surface of bone marrow sLromal cell
19

lf anLlk
k
recepLor on Lhe lmmaLure 8 cell recognlzes Lhe proLeln on sLromal cell cell deaLh
Some rare lmmaLure selfreacLlve 8 cells wlLh acLlve 8AC can undergo furLher llghL chaln gene rearrangemenL a
process known as 8ecepLor edlLlng
4 dependenL on Lhe # of allele remalnlng and only llghL chaln C! rearrangemenL
keceptor Ld|t|ng rearranged llghLchaln genes can alLer Lhe speclflclLy of Lhe llghL chaln by replaclng LhaL confers
auLoreacLlvlLy wlLh one LhaL does noL 8esulL no longer recognlze self proLeln
Ant|gen ependent 9hase Act|vat|on and |fferent|at|on
MaLure 8 Cell
4 Lxpress boLh lgM and lgu on Lhe cell surface
4 8oLh lg heavy chalns on each cell have Lhe same v reglon
4 boLh lg heavy chalns are assoclaLed wlLh Lhe same llghL chalns
4 8oLh lgM and lgu have Lhe same speclflclLy
ClrculaLe ln lymph and blood
Carrled Lo Secondary Lymphold Crgan
naive 8 cell See anLlgen (10) 1
P
ls acLlvaLed whlch produce cyLoklne Lo deLermlne wheLher 8 cell swlLches
class or noL 8 cell ls acLlvaLed Class SwlLchlng and afflnlLy maLuraLlon (lncrease afflnlLy over Lhe course of an
lmmune response) lasma cell or Memory cell ls form lasma cell secreLe anLlbodles LhaL goes Lo perlpheral
lymphold organ (le Spleen and lymph nodes)
















20

UNI1 #8 1he Ma[or n|stocompat|b|||ty Comp|ex (MnC)
Iunct|on of MnC Mo|ecu|es 1o blnd and presenL anLlgenlc pepLldes on Lhe surfaces of cells for recognlLlon (blndlng) by
Lhe anLlgenspeclflc LlymphocyLes recepLors (1C8s)
now to 1 Ce||s recogn|ze Ant|gen? 1 cell recepLors recognlzes a pepLlde fragmenL of Lhe proLeln (LplLope)bound wlLhln
Lhe MPC molecule
AnLlbodles blnd anLlgen as eplLopes on Whole proLesln
1 Cells blnd anLlgen LhaL has been processed" as eplLopes presenLed by MPC on an anLlgen presenLlng Cell (AC)
MnC Important
lmmunlLy CellmedlaLed lmmune response complemenL acLlvaLlon
1ransplanLaLlon 8e[ecLlon vs accepLance (88 relaLlve rlsk)
ulsease lnsullndependenL dlabeLes (88 facLor 100 for human MPC u83 / uCW8)
4 Means a person ls 100x more llkely Lo have lnsulln dependenL dlabeLes lf Lhey have Lhls MPC
MnC ene Crgan|zat|on nLA (numan Leukocyte Ant|gens) Chromosome 6
Class ll MPC genes u uM uC u8 all heLerodlmerlc wlLh and chalns
Class l MPC genes A 8 C all heLerodlmerlc buL 2 chaln expressed on dlfferenL chromosome
1A 1 and 1A 2 Cenes locaLed ln Lhe Class ll reglon Lhen encode pepLlde LransporLer (Ag rocesslng)
LM 1 2 Cenes Large Molecular roLeasome Ag rocesslng
Class lll Cenes encode oLher proLelns LhaL funcLlon ln lmmunlLy eg complemenL proLelns (C4 C2 facLor 8)
cyLoklnes (1nC Lumor necrosls facLor and oLhers
MnC ene Crgan|zat|on n2 (Mouse) Chromosome 17
Class ll Cenes l
A
and l
L

Class l genes u L and k
MnC Lxpress|on nLA
Class l
4 Lxpressed on cell membrane (all nucleaLed cells) as a heLerodlmer chaln + 2mlcroglobulln
4 Plghly polymorphlc (chaln)
4 2 mlcroglobulln gene ls noL encoded wlLhln Lhe MPC complex (Cene locaed on Ch 13 for PLA and Ch
2 for P2)
4 MPC1 lnLeracLs wlLh Cu8 on 1 cells
Class ll
4 Lxpressed only on 8 cells macrophages and uendrlLlc cells (rocesslonal AC)
4 PLAu uCu8
Lxpressed on Lhe cell membrane as a heLerodlmer
Are hlghly polymorphlc
lnLeracL wlLh Cu4 of 1 cells
4 PLAuM
ls noL expressed on cell membrane unllke oLher MPC Class ll molecules
lound predomlnanLly ln speclallzed endosomal comparLmenL
Pelps ln pepeLlde blndlng Lo MPC molecules
21

MnC nap|otype when an lndlvldual lnherlLs Lhe alleles encoded by Lhese closely llnked MPC locl as Lwo seLs one from
each parenLs each seL of alleles ls referred Lo as a haploLype
8oLh alleles lnherlLed from parenLs are expressed
Lhe producL of boLh alleles are found on a ll expresslng cells (le expresslon ls codomlnanL)
Mouse Lxample
4 1here are 6 dlfferenL MPC1 Molecules per cell for Lhe mouse
4 1here are 8 dlfferenL class ll molecules per anLlgen presenLlng cell
4 MaLlng of lnbred ouse sLralns each wlLh a dlfferenL MPC haploLype l1 progeny wlll all have Lhe same
MPC Lype able Lo accepL grafLs from elLher parenLs
4 MaLlng of ouLbred mouse sLralns wlLh dlfferenL MPC haploLypes l1 progeny wlll all have dlfferenL
MPC Lype l1 progeny wlll re[ecL grafLs from elLher parenL
Structure of MnC Mo|ecu|es
Class l Molecule
4 PeLerodlmer of a 43kua chaln noncovalenLly assoclaLed wlLh a 12 kua polypepLlde chaln 2
mlcroglobulln
4 1he 2 mlcroglobulln blnds Lo 3 domaln boLh domalns have Lhe super famlly lmmunoglobulln fold
4 chaln blnds lnLo 3 domalns 1 2 3
4 1 and 2 domalns fold LogeLher lnLo a slngle sLrucLure conslsLlng of 2 segmenLed hellces lylng on a
sheeL of 8 sLrands
Class ll Molecule
4 PeLerodlmer of a 34kua chaln assoclaLed wlLh aL 29kua chaln
4 Lach chaln folds lnLo Lhe domalns LogeLher formlng a compacL 4domaln sLrucLure
4 2 and 2 have Lhe super famlly lmmunoglobulln fold
4 1he 2 domalns formlng Lhe pepLlde blndlng clefL are conLrlbuLed by Lwo dlfferenL chalns noncovalenL
complex
epLlde 8lndlng ClefL
4 Class l ends of bound pepLlde burled ln Class l pepLlde slze 810AA
4 Class ll ends of bound pepLlde are noL burled ln class ll pepLlde slze 1318 AA
1he pepLldeblndlng clefL of class 2 molecules ls open aL boLh ends and Lhe pepLlde lles ln an
elongaLed conformaLlon wlLhln Lhe clefL
|str|but|on of MnC Mo|ecu|es
Class l
4 expressed on all nucleaLed cells
4 MosL hlghly expressed ln hemaLopoleLlc cells (blood cells)
4 8lnd pepLldes derlved from Lndogenously synLheslzed proLelns ln Lhe cell (le vlral proLeln from vlral
lnfecLed cell)
Class ll
4 Lxpresslon ls resLrlcLed Lo anLlgenpresenLlng cells (le 8 cells macrophages uendrlLlc cells) buL also on
acLlvaLed human 1cells
4 8lnd pepLldes derlved from exogenous proLelns LhaL have enLered Lhe cell by hagocyLosls or
endocyLosls

22

9o|ymorph|sm of MnC Mo|ecu|es varlablllLy aL a gene locus ln whlch Lhe varlanLs (mulLlple alleles) occur aL a frequency
of greaLer Lhan 1 (approx 100 dlfferenL alleles for each locus)
le mulLlple alleles of Lhe same gene wlLhln Lhe populaLlon as a whole
1he large amounLs of polymorphlsm of MPc sLems from Lhe hlgh number of alleles
4 60 PLAA 110 PLA8 40 PLAC
varlaLlons beLween alleles resulL ln dlfferences ln AA sequences usually ln Lhe Ag clefL dlverslLy ln Ag blndlng
Pypervarlable reglons poslLlon of amlno acld resldues havlng hlgh varlablllLy
4 varlaLlon ls nC1 random buL locallzed Lo speclflc reglons
Class l amlnoLermlnal domalns of 1 and 2
Class ll amlnoLermlnal 1 and 1 domalns
4 SLable dlverslLy (no hypermuLaLlon)
4 MPC has Lhe MsoL polymorphlc genes presenL ln Lhe genome of every specles analyzed
Slgnlflcance
4 AffecLs anLlgen recognlLlon by 1 cells lndlrecLly by conLrolllng pepLlde blndlng
4 AlloreacLlve 1 cells (1 cells LhaL recognlze nonself MPC) are very abundanL
4 exLends Lhe range of anLlgens Lo whlch Lhe lmmune sysLem can respond
4 ulrecLly affecLs anLlgen recognlLlon by 1 cells (Lhe affecL ls called MPC resLrlcLlon)
MnC kestr|ct|on Comblned effecL of MPC pepLlde blndlng + dlrecL conLacL b/w Lhe MPC molecule and Lhe 1 Cell
recepLor
1 cell resLrlcLed" Lo blndlng anLlgen only when lL ls presenLed ln an MPC l or ll
uual SpeclflclLy of 1Cell 8ecognlLlon of AnLlgen
4 8ecognlzes polymorphlc resldues of selfMPC molecules whlch accounLs for Lhelr MPC resLrlcLlon
4 8ecognlzes resldes of pepLlde anLlgens dlsplayed by Lhese MPC molecules whlch are responslble for
Lhelr anLlgen speclflclLy












23

UNI1 #9 Ant|gen 9rocess|ng and 9resentat|on
Ce||u|ar Compartments
1here are Lwo ma[or comparLmenLs wlLhln Lhe cell separaLed by membranes
CyLosol and veslcular SysLem
8ases of Ant|gen kecogn|t|on
1Cell recognlLlon of anLlgen 8equlres pepLldes derlved from forelgn anLlgens be dlsplayed wlLhln Lhe clefL of an
MPC molecule on Lhe cell membrane
AnLlgen rocesslng lnvolves Lhe sequence of evenLs of proLeln anLlgen degradaLlon and formaLlon of pepLlde
MPC complex
AnLlgen resenLaLlon lnvolves pepLldeMPC complexes LransporLed Lo membrane where Lhey are dlsplayed
1Ce|| |nteract|ons w|th pept|deMnC comp|ex
1 Cell recepLor lnLeracLs wlLh polymorphlc reslde of MPC and Anchor resldue of epLlde
1 Cell conLacL also resldue of pepLlde
uual recognlLlon conLacL wlLh resldues of MPC and anLlgen pepLlde
Ce||s that Iunct|on |n Ant|gen 9resentat|on
rofesslonal AnLlgen resenLaLlon Cells (AC)
4 uendrlLlc Cells (Lhe besL) consLlLuLlvely express MPC ll and cosLlmulaLory 87 molecule
4 Macrophages musL be acLlvaLed Lo express MPC ll and 87 molecules
4 8Cells consLlLuLlvely express MPC ll buL musL be acLlvaLed Lo express 87 molecule
CrlLlcal for lnLeracLlon w/ Cu28 on 1cell acLlvaLes 1 cells + cyLoklne recepLor
nonrofesslonal AC usually doesn'L express class ll
4 llbroblasL (Skln)
4 1hymlc eplLhellal cells
4 vascular endoLhellal cells
4 ancreaLlc beLa cells
4 1hyrold eplLhellal cells
2 9rocess|ng 9resentat|on 9athways
CyLosollc aLhway Lndogenous anLlgens Class l MPC molecules
4 AcLlvaLes CyLoLoxlc 1 cells (Cu*+)
4 eg vlrus lnfecLed cells Lumor cells
LndocyLlc aLhway Lxogenous anLlgens Class ll MPC molecules
4 AcLlvaLes Pelper 1 cells (Cu4+)
4 Ag Laken up from ouLslde cell by macrophages MonocyLes 8 cells dendrlLlc cells and langerhans cells
Cytoso||c 9athway Cell 8lology of AnLlgen orcessln
1 uegradaLlon of proLelns by roLeasome
a cyLosollc proLelns are degraded by a large mulLlcaLalyLlc proLease complex of 28 subunlLs called
proLeasome or LM (low molecularmass polypepLlde)
b roLeln Lo be degraded ls flrsL Lagged by a ublqulLln vla A1
24

2 1ransporLers AssoclaLed wlLh AnLlgen rocesslng (1A1 2)
a boLh 1ap 1 and 1ap 2 genes encode 1 hydrophoblc and one A1 blndlng domaln each and Lhe domalns
are assembled lnLo a heLerodlmer Lo form a LransporLer beLween Lhe L8 membrane
b MuLaLlons ln elLher 1ap 1 or 1A 2 genes adversely affecL anLlgen presenLaLlon by MPC class l molecules
l Cllnlcally paLlenLs have sympLoms of auLolmmunlLy wlLh skln leslons chronlc slnuslLls and
chronlc bacLerlal lnfecLlons of Lhe lungs
c 1ap resulLs ln normal anLlgen presenLaLlon and class l MPC expresslon
3 ln Lhe L8 newly synLheslzed MPC class l chaln rapldly blnd Lo Calnexln (a molecular chaperone) whlch reLalns
chaln ln a parLlally folded sLaLe ln L8 unLll 2mlcroglobulln blnds
4 MPC Class l 2m complex ls released from calnexln and blnds Lo chaperone proLesln CalreLlculln Lrp37) and
1apsln (1A1 assoclaLed proLeln)
a Lrp 37 a proLeln dlsulflde lsomerlse LhaL plays role ln breaklng and reformlng Lhe dlsulflde bond lnL eh
MPC class l 2 durlng pepLlde loadlng
3 epLldes from degraded proLeln ln cyLosol are LransporLed lnLo L8 vlaL eh 1ap LransporLer
6 lormaLlon of MPCepLlde Complex
a 1A dellvers a pepLlde LhaL blnds Lhe MPC complex
b epLlde blndlng compleLes MPC class l foldlng releaslng CalreLlculln
c Cnly folded molecules are exporLed Lo Colgl apparaLus