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Section 4Section 4
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• An enormous family of related, but nAn enormous family of related, but nonidentical glycoproteinsonidentical glycoproteins
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Expression of IgExpression of Ig
• membrane Ig, mIg : expressen on the smembrane Ig, mIg : expressen on the surfaces of resting B lymphocytes, as reurfaces of resting B lymphocytes, as receptorceptor
• secreted Ig, sIg : as antibody(Ab)secreted Ig, sIg : as antibody(Ab)
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Major functions of IgMajor functions of Ig
• Binding AgBinding Ag• Secondary biologic activitiesSecondary biologic activities
– OpsoninsOpsonins– Activate the complement cascadeActivate the complement cascade– Cross the placental barrierCross the placental barrier
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Chapter 1Chapter 1 organization of organization of immunoglobulinsimmunoglobulins
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Molecular structure of IgMolecular structure of Ig
• An enormous family of related, but nonideAn enormous family of related, but nonidentical glycoproteinsntical glycoproteins
• Ab are bifuctional molecules: Ab are bifuctional molecules:
bind specifically to Agbind specifically to Ag
secondary phenomenasecondary phenomena
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The four-chain basic unitThe four-chain basic unit
Symmetrical structureSymmetrical structureTwo Heavy (H) chains, about twice aTwo Heavy (H) chains, about twice a
s large as the L chain s large as the L chain Two Light (L) chains, the smallerTwo Light (L) chains, the smallerCovalent interchain disulfide bridgeCovalent interchain disulfide bridge
ss
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(Ag-biding site)
L chain
H chain(,,,,)
V region
C region
HH22LL22
hinge regiohinge regionn
2 regions2 regions
κ/λ (2:1)
Monomer of Ig
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domainsdomains
Both H and L chains contain folded gloBoth H and L chains contain folded globular domainsbular domains
Each domains has a single intrachain Each domains has a single intrachain disulfide bonddisulfide bond
L chains always contain two domains L chains always contain two domains H chains contain either 4 or 5 domainsH chains contain either 4 or 5 domains
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Immunoglobulin variable Immunoglobulin variable regions and constant regionsregions and constant regions
variable region (V region)variable region (V region)
The sequences of the N-The sequences of the N-terminal terminal
domain in a heavy- or light-chain domain in a heavy- or light-chain
polypeptide are variablepolypeptide are variable– VVHH
– VVL L
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constant region (C region)constant region (C region)
• The sequences of the other The sequences of the other domains remain relatively domains remain relatively constantconstant– CCHH
– CCL L
• L-chain polypeptides contain L-chain polypeptides contain only a Conly a CL L domaindomain
• H-chain polypeptides contain 3 H-chain polypeptides contain 3 or more Cor more CH H domainsdomains
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(二)可变区和恒定区
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The VThe VHH is juxtaposed with the V is juxtaposed with the VL L to fto form an Ag-binding siteorm an Ag-binding site
Each Each monomer of Ig contains two monomer of Ig contains two AAg-binding sitesg-binding sites
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Hinge regionHinge region
Located between the CH1 and CH2 oLocated between the CH1 and CH2 of the H chains in most Igf the H chains in most Ig
Permits flexibility between the two Permits flexibility between the two Fab arms of AbFab arms of Ab
Allows the two arms to open and cloAllows the two arms to open and close to accommodate binding to two se to accommodate binding to two epitopesepitopes
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Flexibility of immunoglobulins
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Hypervariable regionsHypervariable regions
The V regions contain two regions:The V regions contain two regions:
Framework regions----relatively invarFramework regions----relatively invariantiant
Hypervariable regions----extremely vHypervariable regions----extremely variant ariant Complementarity-determining rComplementarity-determining regions (CDRs)egions (CDRs)
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Complementarity-determining regionComplementarity-determining regions (CDRs)s (CDRs)
Each VH and VL contains 3 CDRsEach VH and VL contains 3 CDRs
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Structure of the Structure of the Variable DomainVariable Domain
Relation to AgRelation to Ag--bindingbinding
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Antigen-binding Can Take Many Forms – Best Method
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J chainJ chain IgM and IgA generally exist as polymIgM and IgA generally exist as polym
ersers
Joint Joint monomers to form polymers
n(Hn(H22LL22)) n(monomer)n(monomer)
J chainJ chain (H(H22LL22)n)n JJ
polymerpolymer
J chain and secretary J chain and secretary componentcomponent
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Secretory componentSecretory component
• Associated only with IgA and is founAssociated only with IgA and is found almost exclusively in body secretid almost exclusively in body secretionsons
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Enzymatic digestion products of IgEnzymatic digestion products of Ig
Be most susceptible to cleavage near Be most susceptible to cleavage near the hinge regionthe hinge region• Enzyme papainEnzyme papain• Enzyme pepsinEnzyme pepsin
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Enzyme papainEnzyme papain
Splits an Ig into 3 fragments of Splits an Ig into 3 fragments of roughly similar sizeroughly similar size
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Fab regionFab region: each Fab is monovalent with r: each Fab is monovalent with respect to Ag-binding activityespect to Ag-binding activity
Fc regionFc region: : be recognized by the Fc receptors found be recognized by the Fc receptors found
on many types of cellson many types of cellsDetermine most of the secondary biologiDetermine most of the secondary biologi
c properties of Igc properties of Ig
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Enzyme pepsinEnzyme pepsin
Splits an Ig into:Splits an Ig into:A single large fragment ----F(ab)’2 A single large fragment ----F(ab)’2
fragmentfragmentPFc’PFc’
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F(ab)’2 fragment:F(ab)’2 fragment: • roughly corresponds to 2 disulfidelinked roughly corresponds to 2 disulfidelinked
Fab fragmentsFab fragments• Has divalent Ag-binding activityHas divalent Ag-binding activity
PFc’:PFc’:• Extensively degradedExtensively degraded• Usually does not survive as an intact fragUsually does not survive as an intact frag
mentment
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Hydrolytic fragment of IgHydrolytic fragment of Ig
2 Fab + 1Fc1 F(ab’)2 +pFc’
(papain)(pepsin)
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Chapter 2Chapter 2 Ig classes and Ig classes and subclassessubclasses
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Ig classes, subclasses and typesIg classes, subclasses and types
• Classes and subclasses are Classes and subclasses are determined on the basis of CHdetermined on the basis of CH
• According to CH sequences and According to CH sequences and their physical and biologic their physical and biologic propertiesproperties
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ClassesClasses
• Humans express five different classHumans express five different classes of Ig heavy chainses of Ig heavy chains
• All of the heavy chains in any given IAll of the heavy chains in any given Ig are identicalg are identical
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The classes of heavy chainsThe classes of heavy chains
γ μ α δ εγ μ α δ ε
The classes of Ig containing these CHThe classes of Ig containing these CH
IgG IgM IgA IgD IgEIgG IgM IgA IgD IgE
The The μ and ε contain 5 CHμ and ε contain 5 CH
The γ, α and δ contain 4 CHThe γ, α and δ contain 4 CH
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Subclasses or subtypesSubclasses or subtypes
• IgG1, IgG2, IgG3, IgG4IgG1, IgG2, IgG3, IgG4
• IgA1, IgA2IgA1, IgA2
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All light chains can be classified into 2 All light chains can be classified into 2 types according to CL sequencetypes according to CL sequence• Kappa (κ)Kappa (κ)• lambda (λ)lambda (λ)
No known functional differences exisNo known functional differences exist between two typst between two typs
An Ab molecule has either 2 κ or 2 λ liAn Ab molecule has either 2 κ or 2 λ light chains, but never one of eachght chains, but never one of each
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Isotypes, allotypes and idiotypes Isotypes, allotypes and idiotypes of Igof Ig• Ig are proteinsIg are proteins• Ig can be immunogenicIg can be immunogenic
– Isotypic----isotypeIsotypic----isotype– Allotypic----allotypeAllotypic----allotype– Idiotypic----idiotypeIdiotypic----idiotype
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isotypesisotypesThe antigenic features of a class of IThe antigenic features of a class of I
g Hg HThe genes for isotypic variantThe genes for isotypic variant
s are present in all healthy s are present in all healthy membersmembers
isotypeisotype
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allotypesallotypesVary among individuals within Vary among individuals within
a species and are under a species and are under genetic controlgenetic control
allotypesallotypes
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idiotypesidiotypes
• The unique V-region amino acid seqThe unique V-region amino acid sequences of the homogeneous Ig moleuences of the homogeneous Ig molecules produced by a single cules produced by a single B cell cloneB cell clone
idiotypeidiotype
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Membrane and secreted IgMembrane and secreted Ig• membrane Ig, mIg : expressen on thmembrane Ig, mIg : expressen on th
e surfaces of resting B lymphocytes, e surfaces of resting B lymphocytes, as receptoras receptor
• secreted Ig, sIg : secreted Ig, sIg :
as antibody(Ab)as antibody(Ab)
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Chapter 3Chapter 3 biological activities biological activities
of Igof Ig
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Activities of variable regionsActivities of variable regions
Bind to Ag in vivo or in vitro---resistant to infection by:
Opsonizing (coating) organismsBind to viruses, prevent invationNeutralize toxins of microorganisms
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返回
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Neutralization By Antitoxin Antibodies
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Neutralization By Antiviral Antibodies
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Bacterial ‘Neutralization’ By Ab
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Activities of constant regionsActivities of constant regions
• Complement fixationComplement fixation• OpsonizationOpsonization• ADCCADCC
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Complement fixationComplement fixation
Ag-bound IgM and IgG can initiate tAg-bound IgM and IgG can initiate the classical pathway of complemenhe classical pathway of complement activationt activation
Activated complement mediated lysActivated complement mediated lysis of the pathogen (or other cell)is of the pathogen (or other cell)
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Opsonization Opsonization IgG is an opsonizing Ab Fc confers the opsonizing propertyMany phagocytic cells bear receptor
s of the Fc region of IgG• Macrophages• Polymorphonuclear phagocytes• phagocyte
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调理作用OpsonizationOpsonization
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ADCCADCC
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FcRIII CD16
Antibody Marks Target Cells For NK Cell Attack (ADCC)
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Chapter 4Chapter 4 biological properties anbiological properties an
d functions of Ig classesd functions of Ig classes
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IgGIgG
About 75% of the total serum Ig in normaAbout 75% of the total serum Ig in normal adultsl adults
The most abundant Ab produced during sThe most abundant Ab produced during secondary IRecondary IR
The only class of Ig that can cross the plaThe only class of Ig that can cross the placentacenta
Be capable of fixing serum complementBe capable of fixing serum complement Bind the Fc receptorsBind the Fc receptors
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IgAIgA• In secretions: SIgA ---- dimerIn secretions: SIgA ---- dimer
• On B cell surfaces On B cell surfaces monomermonomer• In the bloodIn the blood
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The predominant Ig produced by B cThe predominant Ig produced by B cells in submucosal lymphoid tissuesells in submucosal lymphoid tissues
The most abundant Ab class found iThe most abundant Ab class found in secretions (milk, bronchial secretin secretions (milk, bronchial secretions, intestinal mucus)ons, intestinal mucus)
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IgMIgM
Pentamer in serumPentamer in serumPredominate in early primary immunPredominate in early primary immun
e responsese responsesThe most efficient complement fixing The most efficient complement fixing
IgIgExpressed on the surfaces of B cells, Expressed on the surfaces of B cells,
particularly naïve B cellsparticularly naïve B cells
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Can be synthesized during the late eCan be synthesized during the late embryo stagembryo stage
The first Ig produced in primary immThe first Ig produced in primary immune responsesune responses
Natural blood group antibodyNatural blood group antibody
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IgDIgD
The physiologic function of IgD is unThe physiologic function of IgD is unknownknown
Rarely secreted in significant amouRarely secreted in significant amounts in bloodnts in blood
Commonly found on the surfaces of Commonly found on the surfaces of B cellsB cells
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mIgM mIgD
成熟 B 细胞 活化 B 细胞未成熟 B 细胞
mIgD can bind Ag and transmit signmIgD can bind Ag and transmit signals to the cell interiorals to the cell interior
When B cells containing mIgD becoWhen B cells containing mIgD become activated, mIgD expression ceasme activated, mIgD expression ceaseses
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IgEIgERepresents only a minute fraction oRepresents only a minute fraction o
f all serum Abf all serum AbHas central involvement in allergic Has central involvement in allergic
disordersdisorders In monomeric formIn monomeric form
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FcRI
IgE Antibody Binds To Mast Cells & Basophils To Arm Them For Mediator Release
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Chapter 5Chapter 5
Ab technologyAb technology
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Poly-clonal abtibody, PcAb The antiserum from an animal The antiserum from an animal immunized against an immunogen is aimmunized against an immunogen is a complex mixture of structurally diverse complex mixture of structurally diverse Abs recognizing multiple epitopes on Abs recognizing multiple epitopes on the immunogen, and its composition the immunogen, and its composition fluctuates unpredictably over the life of fluctuates unpredictably over the life of the animal the animal
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带有多个表位
抗原
针对不同表位的 B 细胞
克隆增殖
多种抗体的混合物
(多克隆抗体)
刺激小鼠
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Mono-clonal antibody,McAbMono-clonal antibody,McAb
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