1、Immunoglobulin Structure-Function Relationship,免疫球蛋白的结构与功能的关系,Signalling antigen receptors on B cells - bifunctional antigen-binding secreted molecules（B 细胞表面受体和分泌的抗体） Structural conservation and infinite variability - domain structure（结构上不仅保守而且无限可变的）. The Immunoglobulin Gene Superfamily （免疫球蛋白的超家族）
2、 The immunoglobulin fold （免疫球蛋白的折叠） Framework and complementarity determining regions - hypervariable loops （框架结构和可变区） Modes of interactions with antigens （与抗原相互作用的模型） Effector mechanisms and isotype role of the Fc. （Fc 区的作用） Multimeric antibodies and multimerisation Characteristics and properties o
3、f each Ig isotype Ig receptors and their functions,Immunoglobulin Structure-Function Relationship,Cell surface antigen receptor on B cells B 细胞表面受体和分泌的抗体 Allows B cells to sense their antigenic environment Connects extracellular space with intracellular signalling machinery Secreted antibody （抗体）Neu
4、tralisation （中和作用）Arming/recruiting effector cells （激活或者诱导功能细胞）Complement fixation （帮助机体对抗原的清除）,Immunoglobulin Structure-Function Relationship,Immunoglobulins are Bifunctional Proteins,Immunoglobulins must interact with a small number of specialised molecules - （免疫球蛋白必须与特殊分子相互作用）Fc receptors on cell
5、s （细胞表面的Fc受体）Complement proteins （辅助蛋白）Intracellular cell signalling molecules （细胞内信号转导分子） whilst simultaneously recognising an infinite array of antigenic determinants. （同时能够识别无限抗原族）,Structural conservation and a capacity for infinite variability in a single molecule is provided by a DOMAIN structu
6、re. （结构上不仅保守而且无限可变的- 抗体结构域） Ig domains are derived from a single ancestral gene that has duplicated, diversified and been modified to endow an assortment of functional qualities on a common basic structure（Ig结构域源于一个原始基因，复制，多元化，修饰等） Ig domains are not restricted to immunoglobulins （Ig 结构域不仅仅局限于免疫球蛋白）
7、. The most striking characteristic of the Ig domain is a disulphide bond - linked structure of 110 amino acids long（Ig结构域最明显的特点是其双硫键，连接了110个氨基酸）.,Immunoglobulin domains,The genes encoding Ig domains are not restricted to Ig genes. Although first discovered in immunoglobulins, they are found in a sup
8、erfamily of related genes, particularly those encoding proteins crucial to cell-cell interactions and molecular recognition systems. IgSF molecules are found in most cell types and are present across taxonomic boundaries,Ig gene superfamily - IgSF,Antibodies are Proteins that Recognize Specific Anti
9、gens 抗体能够特异性的识别抗原,Epitopes（抗原决定簇 ）: Antigen Regions that Interact with Antibodies,Consequences of Antibody Binding 抗体结合效应,Domains are folded, compact, protease resistant structures,Domain Structure of Immunoglobulins 免疫球蛋白的结构域,Light chain C domains k or l,Heavy chain C domains a, d, e, g, or m,CH3,C
10、H3,CH2,CH3,CH2,CH1,CH3,CH2,CH1,VH1,CH3,CH2,CH1,VH1,VL,CH3,CH2,CH1,VH1,CL,VL,CH3,CH2,CH1,VH1,CL,VL,Hinge,CH3,CH2,CH1,VH1,VL,CL,Elbow,Fv,Flexibility and motion of immunoglobulins,Hinge,Fv,Fb,Fab,CH3,CH2,CH1,VH1,VL,CL,Fc,Elbow,Carbohydrate,The Immunoglobulin Fold,The characteristic structural motif of
11、all Ig domains,Unfolded VL region showing 8 antiparallel b-pleated sheets connected by loops.,The Immunoglobulin Fold,Immunoglobulins must interact with a finite number of specialised molecules - Easily explained by a common Fc region irrespective of specificity - whilst simultaneously recognising a
12、n infinite array of antigenic determinants.,In immunoglobulins, what is the structural basis for the infinite diversity needed to match the antigenic universe?,Immunoglobulins are Bifunctional Proteins,Cytochromes C,Variability of amino acids in related proteins Wu & Kabat 1970,Distinct regions of h
13、igh variability and conservation led to the concept of a FRAMEWORK (FR), on which hypervariable regions were suspended.,Framework and Hypervariable regions,Most hypervariable regions coincided with antigen contact points - the COMPLEMENTARITY DETERMINING REGIONS (CDRs),Hypervariable CDRs are located
14、 on loops at the end of the Fv regions,Space-filling model of (Fab)2, viewed from above, illustrating the surface location of CDR loops,Light chains Green and brown Heavy chains Cyan and blue CDRs Yellow,The framework supports the hypervariable loops The framework forms a compact b barrel/sandwich w
15、ith a hydrophobic core The hypervariable loops join, and are more flexible than, the b strands The sequences of the hypervariable loops are highly variable amongst antibodies of different specificities The variable sequences of the hypervariable loops influences the shape, hydrophobicity and charge
16、at the tip of the antibody Variable amino acid sequence in the hypervariable loops accounts for the diversity of antigens that can be recognised by a repertoire of antibodies,Hypervariable loops and framework: Summary,Antigens vary in size and complexity,Protein: Influenza haemagglutinin,Hapten: 5-(
17、para-nitrophenyl phosphonate)-pentanoic acid.,Antibodies interact with antigens in a variety of ways,Antigen inserts into a pocket in the antibody,Antigen interacts with an extended antibody surface or a groove in the antibody surface,Fv,Flexibility and motion of immunoglobulins,60 weakly neutralisi
18、ng McAb Fab regions,EBV coated with a corona of anti-EBV antibodies,EBV coated with antibodies and activated complement components,Antibody + complement- mediated damage to E. coli,Non-covalent forces in antibody - antigen interactions,Electrostatic forces Attraction between opposite charges Hydroge
19、n bonds Hydrogens shared between electronegative atoms Van der Waals forces Fluctuations in electron clouds around molecules oppositely polarise neighbouring atoms Hydrophobic forces Hydrophobic groups pack together to exclude water (involves Van der Waals forces),Why do antibodies need an Fc region
20、?,Detect antigen Precipitate antigen Block the active sites of toxins or pathogen-associated molecules Block interactions between host and pathogen-associated molecules,The (Fab)2 fragment can -,Inflammatory and effector functions associated with cells Inflammatory and effector functions of compleme
21、nt The trafficking of antigens into the antigen processing pathways,but can not activate,Structure and function of the Fc region,Fc structure is common to all specificities of antibody within an ISOTYPE (although there are allotypes),The structure acts as a receptor for complement proteins and a lig
22、and for cellular binding sites,Monomeric IgM,IgM only exists as a monomer on the surface of B cells,Cm4 contains the transmembrane and cytoplasmic regions. These are removed by RNA splicing to produce secreted IgM,Monomeric IgM has a very low affinity for antigen,Cm3 binds C1q to initiate activation
23、 of the classical complement pathway Cm1 binds C3b to facilitate uptake of opsonised antigens by macrophages Cm4 mediates multimerisation (Cm3 may also be involved),Polymeric IgM,IgM forms pentamers and hexamers,Multimerisation of IgM,1. Two IgM monomers in the ER(Fc regions only shown),2. Cysteines
24、 in the J chain form disulphide bonds with cysteines from each monomer to form a dimer,3. A J chain detaches leaving the dimer disulphide bonded.,4. A J chain captures another IgM monomer and joins it to the dimer.,5. The cycle is repeated twice more,6. The J chain remains attached to the IgM pentam
25、er.,Antigen-induced conformational changes in IgM,Planar or Starfish conformation found in solution. Does not fix complement,Staple or crab conformation of IgM Conformation change induced by binding to antigen. Efficient at fixing complement,IgM facts and figures,Heavy chain: m - Mu Half-life: 5 to
26、10 days % of Ig in serum: 10 Serum level (mgml-1): 0.25 - 3.1 Complement activation: + by classical pathway Interactions with cells: Phagocytes via C3b receptors Epithelial cells via polymeric Ig receptor Transplacental transfer: No Affinity for antigen: Monomeric IgM - low affinity - valency of 2 P
27、entameric IgM - high avidity - valency of 10,IgD facts and figures,IgD is co-expressed with IgM on B cells due to differential RNA splicing Level of expression exceeds IgM on nave B cells IgD plasma cells are found in the nasal mucosa - however the function of IgD in host defence is unknown - knocko
28、ut mice inconclusive Ligation of IgD with antigen can activate, delete or anergise B cells Extended hinge region confers susceptibility to proteolytic degradation,Heavy chain: d - Delta Half-life: 2 to 8 days % of Ig in serum: 0.2 Serum level (mgml-1): 0.03 - 0.4 Complement activation: No Interactio
29、ns with cells: T cells via lectin like IgD receptor Transplacental transfer: No,IgA dimerisation and secretion,IgA is the major isotype of antibody secreted at mucosal sufaces Exists in serum as a monomer, but more usually as a J chain-linked dimer, that is formed in a similar manner to IgM pentamer
30、s.,IgA exists in two subclasses IgA1 is mostly found in serum and made by bone marrow B cells IgA2 is mostly found in mucosal secretions, colostrum and milk and is made by B cells located in the mucosae,Secretory IgA and transcytosis,Stalk of the pIgR is degraded to release IgA containing part of th
31、e pIgR - the secretory component,IgA facts and figures,Heavy chains: a1 or a2 - Alpha 1 or 2 Half-life: IgA1 5 - 7 days IgA2 4 - 6 days Serum levels (mgml-1): IgA1 1.4 - 4.2 IgA2 0.2 - 0.5 % of Ig in serum: IgA1 11 - 14 IgA2 1 - 4 Complement activation: IgA1 - by alternative and lectin pathway IgA2
32、- No Interactions with cells: Epithelial cells by pIgR Phagocytes by IgA receptor Transplacental transfer: No,To reduce vulnerability to microbial proteases the hinge region of IgA2 is truncated, and in IgA1 the hinge is heavily glycosylated. IgA is inefficient at causing inflammation and elicits pr
33、otection by excluding, binding, cross-linking microorganisms and facilitating phagocytosis,IgE facts and figures,IgE appears late in evolution in accordance with its role in protecting against parasite infections Most IgE is absorbed onto the high affinity IgE receptors of effector cells IgE is also
34、 closely linked with allergic diseases,Heavy chain: e - Epsilon Half-life: 1 - 5 days Serum level (mgml-1): 0.0001 - 0.0002 % of Ig in serum: 0.004 Complement activation: No Interactions with cells: Via high affinity IgE receptors expressed by mast cells, eosinophils, basophils and Langerhans cells
35、Via low affinity IgE receptor on B cells and monocytes Transplacental transfer: No,The high affinity IgE receptor (FceRI),The IgE - FceRI interaction is the highest affinity of any Fc receptor with an extremely low dissociation rate. Binding of IgE to FceRI increases the half life of IgE Ce3 of IgE
36、interacts with the a chain of FceRI causing a conformational change.,IgG facts and figures,Heavy chains: g 1 g 2 g3 g4 - Gamma 1 - 4 Half-life: IgG1 21 - 24 days IgG2 21 - 24 days IgG3 7 - 8 days IgG4 21 - 24 days Serum level (mgml-1): IgG1 5 - 12 IgG2 2 - 6 IgG3 0.5 - 1 IgG4 0.2 - 1 % of Ig in seru
37、m: IgG1 45 - 53 IgG2 11 - 15 IgG3 3 - 6 IgG4 1 - 4 Complement activation: IgG1 + IgG2 + IgG3 + IgG4 No Interactions with cells: All subclasses via IgG receptors on macrophages and phagocytes Transplacental transfer: IgG1 + IgG2 + IgG3 + IgG4 +,Fcg receptors,Receptor Cell type Effect of ligation FcgR
38、I Macrophages Neutrophils, Eosinophils, Dendritic cells Uptake, Respiratory burst FcgRIIA Macrophages Neutrophils, Eosinophils, Platelets Langerhans cells Uptake, Granule release FcgRIIB1 B cells, Mast Cells No Uptake, Inhibition of stimulation FcgRIIB2 Macrophages Neutrophils, Eosinophils Uptake, I
39、nhibition of stimulation FcgRIII NK cells, Eosinophils, Macrophages, Neutrophils Mast cells Induction of killing (NK cells),High affinity Fcg receptors from the Ig superfamily:,The neonatal Fcg receptor,The FcgRn is structurally related to MHC class I In cows FcgRn binds maternal IgG in the colostru
40、m at pH 6.5 in the gut. The IgG receptor complex is trancytosed across the gut epithelium and the IgG is released into the foetal blood by the sharp change in pH to 7.4 Some evidence that this may also happen in the human placenta, however the mechanism is not straightforward.,Human FcgRn,Human MHC Class I,
