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Chapter 4 : All about B Cells

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All about B Cells, Page 1 of 2

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Abstract:

This chapter describes the nature of antibodies, their reaction with antigens, how antibodies are produced, and how B cells are activated and differentiate to express different immunoglobulin classes (isotypes). The laboratory techniques used to detect and measure antigen-antibody reactions have been critical tools in the development of immunological knowledge; because of the extreme specificity and sensitivity of antibodies, these techniques have also been widely used in other fields including protein chemistry, molecular biology, forensics, and clinical medicine. B cells express membrane immunoglobulin molecules that can bind to antigen and generate activation signals for B-cell proliferation and differentiation. B-cell receptor (BCR) signaling can be modulated by other cell surface receptors. Two of these are the CR2 receptor for complement, which promotes B-cell activation, and the Fc receptor FcγRIIb, which inhibits signaling. The structure of antibody expressed on the surface membrane of a B lymphocyte differs from that of secreted antibody in that additional amino acids are present on the C terminus. The most unusual feature of immunoglobulin structure is the vast diversity of antibody molecules with different antigen-binding specificities. Unique genetic mechanisms for generating antibody diversity were discovered when the genes encoding homogeneous myeloma proteins were compared with the corresponding sequences in germ line DNA. More recently, monoclonal antibodies have been proposed for the diagnosis and treatment of cancer. Therapeutic approaches involve both antibodies tagged with radioactivity to provide localized radiotherapy and antibodies tagged with various toxins.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.1
Figure 4.1

Demonstration of antibodies in the gamma globulin fraction of serum. If serum is placed under an electric field, the proteins will migrate in the charged gradient. The black line indicates the levels of proteins in the migration pattern after absorption with antigen. The blue-green line indicates the levels before absorption. By this method, the serum antibodies were shown to be the least negatively charged (gamma globulin).

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.2
Figure 4.2

Equilibrium dialysis. A solution of antibody is placed inside a dialysis bag, and the bag is placed in a solution of free hapten. The dialysis bag is chosen to be permeable to the hapten but not to the antibody. Free hapten will dialyze into the bag, where it will be bound by larger antibody molecules. When the system reaches equilibrium, the concentration of hapten inside the bag will be equal to the concentration of free hapten outside the bag, plus the concentration of the hapten bound to the antibody inside the bag.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.3
Figure 4.3

Protease fragments of IgG. IgG molecules are cleaved by proteases to generate fragments that were used to determine the structure of IgG. The intact IgG molecule is indicated in the center. The heavy chain is indicated as a bluegreen bar, and the light chain is indicated as a shorter white bar; in each case the N-terminal V domain is stippled, and the C domains are not. The thin black bars represent the disulfide bonds that link the heavy and light chains. Cleavage by papain produces three fragments: two identical fragments, the Fab fragments, can bind antigen but cannot cross-link the antigen (are univalent); and one, the Fc fragment, contains the C regions of the heavy chain. Cleavage by pepsin produces one fragment and many small fragments. The one large fragment consists of two Fab fragments linked by disulfide bonds because the pepsin cleaves the heavy chains on the C-terminal side of the disulfide bonds linking the two heavy chains. The Fc region is cleaved into small pieces.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.4
Figure 4.4

Schematic view of the four-chain structure of a human IgG molecule. Numbers on the right side indicate actual residues of a myeloma protein EU. Numbers of Fab fragments on the left side are aligned for maximum homology: light chains numbered according to E. A. Kabat (J. Immunol. 125:961, 1980). Immunoglobulin domains are shown by the four “loops” formed by intrachain disulfide bonds (SS). VL and VH, light- and heavy-chain V domains. CH1, CH2, and CH3, domains of the C region of the heavy chain. CL, C region of the light chain. Hinge region in which two heavy chains are linked by disulfide bonds is indicated by the thin colored lines. Hypervariable regions (complementarity-determining regions) are indicated by thicker colored lines. Attachment of carbohydrate is at residue 297. Arrows at residues 107 and 110 denote the transition from V to C regions. Sites of action of papain before the hinge region and of pepsin after the hinge region show why papain produces Fab monomer and pepsin produces F(ab)2 dimers. Locations of heritable allotypic differences (Gm on the heavy chain and Inv on the light chain) are shown.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.5
Figure 4.5

Ribbon diagram of a light chain. This three-dimensional view of the folding of the α carbon chain backbone is a visualization based on X-ray diffraction of immunoglobulin crystals. The V and C domains are rotated 160° with respect to each other. The broad arrows (“ribbons”) show the seven segments of β-pleated sheet structure in each domain, with one face of three strands (blue-green) roughly parallel to another face of four strands. The loops projecting out at the left represent the three complementarity-determining regions (CDR1, CDR2, and CDR3), which, together with three similar loops from the heavy-chain V region, form the antigen-combining site.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.6
Figure 4.6

X-ray crystallographic space-filling three-dimensional model of an IgG molecule. One complete heavy chain is light blue-green, and the other is bluegreen; each light chain is white, and carbohydrate is cross-hatched. The positions of the heavy-chain hinge and the two antigen (Ag) binding sites are labeled.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.7
Figure 4.7

Comparison of immunoglobulin classes in humans. The five isotype classes (IgM, IgD, IgA, IgE, and IgG) contain similar basic subunits composed of two identical heavy chains (known as μ, δ, α, ϵ, and λ) and two identical light chains (either к or λ) attached to each of the heavy chains. There are four human λ subtypes. Disulfide bonds link heavy and light chains, as shown by the thin bluegreen lines, and also link IgA into dimers and IgM into pentamers. IgM pentamers and IgA dimers also contain a 15-kDa J (joining)-chain polypeptide. Two of the five Ig isotypes exert their major function bound to cells through the Fc region of their heavy chains—IgD as a lymphocyte receptor and IgE as an effector molecule on mast cells to which it is bound by the Fcϵ receptor on the mast cell

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.8
Figure 4.8

Variability plot of the V domain of a heavy chain. A parameter reflecting sequence variability at each codon position in the heavy-chain V region shows three clusters of hypervariability. These are known as CDR1, CDR2, and CDR3. The V region of the heavy chain is folded so that the CDRs contact antigen and determine the specificity and, to some extent, the avidity of the binding reaction.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.9
Figure 4.9

Formation of an antigen-binding site (paratope) by folding of the VH regions of the light and heavy chains of an antibody molecule so that the hypervariable regions responsible for binding the epitope are brought together. The numbers refer to amino acid residues. Glycine residues, which are usually present at the positions indicated, are important in chain folding. The hypervariable amino acid regions occur at specific positions in the peptide chain of the VH regions. These “hot spots” lie relatively close together in the antigen-binding site and form a continuous surface capable of providing complementarity with a specific epitope. For a given antibody, not all the hypervariable regions need to be involved with binding of a given antigen.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.10
Figure 4.10

Paratopes, epitopes, idiotypes, and allotypes. The two contact surfaces between antibody and antigen (upper panel) are known as paratope (antigen-binding site) on the antibody and epitope on the antigen. An antibody may be recognized as an antigen by another antibody, and, depending on the part recognized, these are known as anti-idiotypes or antiallotypes. Anti-idiotypes are unique to each antibody and may react with the paratope, with sites adjacent to the paratope or both. Antiallotypes are genetically inherited determinants that are present in different antibodies in the same individual (see below).

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.11
Figure 4.11

The quantitative precipitin reaction. If increasing amounts of antigen are added in separate tubes to constant amounts of antiserum, the amount of precipitate increases to a maximum point and then decreases. This phenomenon results from the nature of the precipitate: a large network of multivalent antigen and divalent antibody forming a lattice structure that becomes insoluble as the mass of the lattice displaces surface ions required to maintain solubility. When either antibody or antigen is in excess, the amount of cross-linking is insufficient to form an insoluble lattice. The concentration ratio of antigen and antibody yielding maximum precipitation is called the “equivalence point.” Determination of the equivalence point by this type of titration can be used to measure the amount of antibody if the antigen concentration is known or to measure the amount of antigen if the antibody concentration is known.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.12
Figure 4.12

Simple gel diffusion, or Oudin, tube. A solution of antigen is placed over agar containing antibody in a tube. A precipitin band will form where the antigen meets the antibody. The precipitin band will move into the agar as more antigen from the solution diffuses into the agar. The distance that the band moves into the agar at any point in time is a function of concentration of the antigen added (higher concentration of antigen results in further diffusion into the agar). By comparison of the distance of migration of bands formed by known concentrations of an antigen with the distance of migration of band produced by solutions containing unknown amounts of the same antigen, an accurate measure of the antigen concentration in the unknown solution can be made.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.13
Figure 4.13

Radial immunodiffusion, a simple method for quantitation using the principle of the equivalence point between antibody and antigen. Antigen is placed in a well in an agar gel containing a known concentration of antibody. As antigen diffuses into the agar, it forms a precipitin ring at the equivalence point. The higher the concentration of the antigen, the larger the precipitin ring will be.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.14
Figure 4.14

Double diffusion in agar (Ouchterlony) technique. If two identical antigen solutions (Ag1) are placed in adjacent agar wells, and an appropriate antiserum (Ab) is placed in an equidistant well, two precipitin bands will form and will fuse (reaction of identity) because the antigen is the same. If two different antigens (Ag1 and Ag2) are placed in adjacent wells, and antibody to both antigens is placed in an equidistant well, the two precipitin lines will cross since the two antigen-antibody precipitations occur independently (reaction of nonidentity). If an antigen with multiple epitopes is placed in one well (Ag1), and a related antigen sharing some, but not all, epitopes is placed in the second well (Ag1 the antibodies directed against the shared epitopes will form a fused precipitation pattern (identity reaction); but antibodies against epitopes of Ag1 missing in Ag1 will not interact with that molecule and will form a “spur,” extending the precipitin line between Ab and Ag1.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.15
Figure 4.15

Immunoelectrophoresis. To visualize the precipitin lines formed between multiple protein antigens and a mixture of antibodies, the antigens are placed in a well cut into an agar gel (Ag) and subjected to a voltage gradient (electrophoresis), which causes the proteins to move into the gel (step I); the final position of each protein (shaded areas) depends on its charge and size. After electrophoresis, the antibody mixture is placed in a linear trough and allowed to diffuse toward the separated protein antigens (step II). An arc-shaped precipitin line is formed at the equivalence point of each antibody-antigen combination.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.16
Figure 4.16

Agglutination of particulate antigens. (A) Divalent antibody forms a lattice with multivalent particulate antigen. (B) Agglutinated particles settle to the bottom of a round-bottomed tube in a diffuse layer, whereas nonagglutinated particles roll down the sides of the tube to form a tight “button” at the bottom. The difference between these patterns is easily seen from below the tube.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.17
Figure 4.17

Radioimmunoassay. Very small amounts of antigen may be accurately measured by the ability of the antigen to compete with radiolabeled antigen binding to antibody. If a mixture of unlabeled and radiolabeled antigen is added to an antibody solution in slight antigen excess, the amount of labeled antigen bound to the antibody is a function of the amount of unlabeled antigen present. On the basis of this competition, a standard curve can be constructed by adding increasing amounts of unlabeled antigen to constant amounts of labeled antigen and antibody. When relatively small amounts of unlabeled antigen are present, all of the radiolabeled antigens will be precipitated. As the amount of unlabeled antigen is increased, less radiolabeled antigen is bound to the antibody. When a standard curve using increasing amounts of known unlabeled antigen has been constructed, the amount of antigen in an unknown solution can be determined by comparing the degree of inhibition of binding of labeled antigen by the unknown antigen solution to that produced by known amounts of the same antigen.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.18
Figure 4.18

Fluorescent antibody techniques for detecting antigens in tissues. In the direct technique, specific antibody is labeled with fluorescent compound and added to tissue sections. The reaction of specific antibody to antigenic sites in the tissue sections is detected by exposing the labeled tissue sections to UV light and visualizing areas that “light up” under a fluorescence microscope. In the indirect technique, unlabeled antibody is incubated with tissue antigen. Fluorescein-labeled antibody to the first antibody is then added. The first antibody provides more antigenic sites for the second antibody to bind than was provided by the tissue antigen, increasing the technique's sensitivity. In the mixed antiglobulin technique, antigens on the first antibody are used to react to binding sites of the second antibody. The second antibody is then labeled by adding fluorescently labeled immunoglobulin of the same species as that of the first antibody. This variation of the technique is especially useful in labeling surface immunoglobulin molecules when the tissue antigen is an immunoglobulin. To identify antibody rather than antigen in tissue sections, a “sandwich” technique is used. Antigen is added to tissue and is bound by the specific antibody in the tissue. Specific fluorescein-labeled antibody to antigen is added and reacts with antigen now bound to the antibody in the tissue.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.19
Figure 4.19

Identification of different cell types in the blood by flow cytometry. (A) In the first passage, preparations of white blood cells that have had red cells removed by lysis are “gated” by forward and side light scattering. Light is scattered by cytoplasmic granules. This identifies those white cells that contain granules (polymorphonuclear leukocytes and monocytes) and separates them from lymphocytes. Each dot represents a cell. The large groups of cells to the right with high side scatter are granulocytes; the group of cells at the top to the left with high forward scatter are monocytes. The cells in the area outlined in blue-green are lymphocytes. (B) The fraction containing the lymphocytes (which do not scatter light) is stained for CD3 (T-cell receptor) and CD4 (T-helper cells). Four quadrants are identified: CD3 CD4, CD3CD4+, CD3+ CD4, and CD3+ CD4+ The first quadrant identifies null cells; the second identifies residual monocytes that are weakly CD4+; the third identifies non-CD4+ T cells, most likely CD8+ T-suppressor cells; and the fourth identifies CD4+ T-helper cells.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.20
Figure 4.20

Fluorescence-activated cell sorting. (1) Fluorescently stained cells are forced out of a small nozzle, forming a stream in which they pass through a laser beam one cell at a time. (2) An optical sensor linked to a computer detects fluorescent characteristics and classifies each cell according to conditions set by the investigator (e.g., light scattering, different fluorescent wave lengths, size). (3) As the stream forms droplets—containing no more than one cell—the computer imparts a positive, negative, or zero charge to each droplet based on the fluorescence wave length emitted by each labeled cell. (4) The charged droplets pass between charged plates, which deflect the droplets into appropriate collection tubes.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.21
Figure 4.21

Immunodetection of proteins on one-dimensional (1D) and two-dimensional (2D) gels. The left panel shows three lanes from a Western blot. The first two lanes show protein bands as detected by a nonspecific protein stain. Lane 3 shows the result of transferring the same set of proteins in lane 2 onto a solid support and then immunostaining with a labeled antibody; a subset of the total proteins is detected (blue-green bands). The right panel shows a similar comparison between proteins detected by nonspecific protein staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (upper panel) and the subset of proteins detected after blotting to a solid support and immunostaining (blue-green spots in lower panel).

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.22
Figure 4.22

Variation of ELISA technology known as a “sandwich” assay, because the antigen is detected sandwiched between two antibody molecules—unlabeled antibody bound to the plate and an enzyme-labeled antibody added after the antigen has reacted with the antibody bound to the plate. After the wells of a microtiter plate are coated with a constant amount of antibody and then “blocked” by absorption of nonspecific proteins, varying known amounts of antigen are added to different wells and allowed to bind to antibody on the plate. An enzyme-linked antibody against the antigen is then added in excess, and unbound antibody is removed by washing. Finally, a solution containing chromogenic substrate is added, and the rate of color production is measured in an automated plate reader; fluorogenic substrates may also be used with fluorescence-detecting plate readers.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.23
Figure 4.23

Immune elimination. A three-stage elimination of diffusible antigen from the bloodstream of previously nonimmunized animals has been observed. Upon intravascular injection of antigen, the blood level of antigen drops rapidly until only about 40% of injected antigen remains in blood. This is due to equilibration of diffusible antigen between intravascular and extravascular fluid compartments (equilibration phase). After this rapid equilibration, antigen is slowly removed by normal metabolic processes (nonimmune catabolism phase) until the onset of antibody production between 7 and 10 days after antigen injection. Appearance of antibody results in rapid elimination of antigen (immune elimination phase) due to formation of antigen-antibody complexes and their removal by the reticuloendothelial system. During the first part of the immune elimination phase, soluble antigen-antibody complexes (formed in antigen excess) may be demonstrated in the blood until there is enough antibody to form insoluble or antibody excess complexes. These soluble complexes are responsible for the lesions of serum sickness (see chapter 10). After antigen is completely removed, free antibody appears in the blood. If antigen is injected into an animal that already has circulating antibody, antigen is removed in one rapid immune elimination phase.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.24
Figure 4.24

V-J rearrangements of κ light-chain genes. In nonlymphoid tissue (germ line) DNA, there are no functional, complete immunoglobulin genes. The two segments that can form a complete κ gene lie in two clusters: the V gene segments and the much smaller cluster of J region segments. A complete functional κ gene is expressed in a lymphocyte after one of the V segments (shown in black) recombines with one of the J regions.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.25
Figure 4.25

Junctional diversity in V gene assembly. The figure shows the DNA sequence of two germ line murine κ elements in the region involved in recombination during V gene assembly: the germ line Vκ41 segment (highlighted in bluegreen) and Jκ1. The next four lines show possible recombination junctions. The first, which was found in the myeloma MOPC41, includes the Vκ41 sequence up to the second nucleotide of codon 95 and then switches to Jκ1 sequence. The next shows the consequences of a recombination junction one nucleotide further downstream; the G→T change in the last nucleotide of codon 95 does not change any encoded amino acids. The last two lines, illustrating two other possible recombination products, are associated with amino acid changes that have been observed in actual murine κ chains. The examples shown maintain the triplet reading frame between V and J, which is required for a functional gene but occurs in only about one-third of recombination events. If the triplet reading frame is not maintained, the rearranged gene will not be expressed, and recombination at the allelic gene will be attempted. If neither allele of the κ-chain gene has a productive rearrangement, rearrangement of λ light-chain gene may be attempted.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Figure 4.26

Recombination signal sequences (RSSs). The conserved 7-mer and 9- mer sequences that target V assembly recombination are shown flanking the elements involved in these rearrangements. The V, J, and D coding regions are depicted as black rectangles. The spacing between the 7-mer and 9-mer is either 12 or 23 bp as indicated. Recombination occurs almost exclusively between pairs of elements with different RSS spacing.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.27
Figure 4.27

Structure of the human λ light-chain locus. Four of the seven Jλ-Cλ regions in the most frequent haplotype are functional (black rectangles), whereas three regions (4, 5, and 6) are pseudogenes (gray rectangles) in almost all individuals. Variant loci containing extra duplications between the second and third λgenes are indicated by the dashed lines. Flanking regions containing λ-related sequences are shown.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.28
Figure 4.28

Splicing of heavy-chain exons in the formation of membrane and secreted immunoglobulin and μ and δ heavy chains. The exons of a rearranged heavy-chain gene are shown, including both μ and δ loci. Sequences that are spliced out at the RNA level are shown as V-shaped lines. In the case of μ heavy chain, all mature mRNAs show removal of the intron separating most of the signal peptide sequence from the V region, the JH-Cμ intron, and the three intra-Cμ domain introns. In μs mRNA, the transcript includes the last two codons of the secreted heavy chain and terminates at a poly(A) addition site (black dot) close downstream.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.29
Figure 4.29

Isotype (Ig class) switch recombination. The human heavy-chain gene locus is spread out over roughly 300 kb and includes two duplication units of γ-γ-ϵ-α genes, of which only the more downstream γ2-γ4-ϵ-α2 unit is shown here. Isotype switching involves DNA deletion events whose endpoints usually fall within the repetitive switch (S) regions that lie 5′ of each C region. In the isotype switch deletion illustrated here, a composite Sμ-Sϵ region is formed by the recombination; as a consequence, the Cϵ gene is moved downstream of VDJ to the position formerly occupied by Cμ. The resulting DNA is a template for transcription of VDJ-Cϵ mRNA that encodes the ϵ heavy chain.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.30
Figure 4.30

Human immunoglobulin heavy-chain gene locus deletions. This map shows the JH regions and the entire C region gene locus. Black rectangles represent C region genes for the nine expressed isotypes (Ig classes): gray rectangles represent two pseudogenes in the locus. The numbers indicate the approximate distances between the C region genes in kilobases. This map order has been deduced from overlapping clones and Southern blotting of large fragments and is confirmed by analysis of natural germ line deletion mutations in humans (shown by the lines underlying the locus).

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.31
Figure 4.31

B-cell differentiation. Shown is the sequence of marker and Ig gene rearrangements during stages of B-cell differentiation. Cells of the B-cell lineage arise from a multipotent hematopoietic stem cell and are first identified by the presence of RAG1,2 and rearrangement of the DJ regions of the μ-chain gene, as well as the early B-cell markers CD19 and B220, and the nonimmunoglobulin components of surface Ig, Igα and Igβ. The pro-B cells share c-Kit, the receptor for stem cell factor, and TdT with more primitive stem cells. In the absence of heavy-chain expression, the pro-B cell expresses segments of the J and C regions of λ surrogate light chains (SLC), which are later expressed on pre-B cells associated with a αμ heavy chain (μSLC). In pre-B cells, rearrangement of the VDJC of the μ-chain gene occurs, and later the VJ segments of the light-chain genes are rearranged. B cells are recognized by the presence of IgM chain on the cell surface, and in the more mature stage in the periphery, by the presence of IgM and IgD. T-cell stimuli (CD40L and cytokines) lead to immunoglobulin class (isotype) switching in germinal centers of peripheral lymphoid tissue (mostly in lymph nodes) and differentiation to plasma cells secreting antibody of only one immunoglobulin class.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Image of Figure 4.32
Figure 4.32

Detection of a V-J rearrangement in a B-cell tumor by Southern blotting. The upper part of the figure shows nonlymphoid (germ line) DNA (left) and resulting Southern blots (right); the lower part of the figure shows the corresponding features of DNA from a myeloma containing a Vκ-Jκ rearrangement. When germ line DNA is digested with a restriction endonuclease (having sites indicated by the arrows), and blots of the resulting DNA fragments are probed for V and C regions, the bands for the V and C regions will be different. After V-J recombination, both V and J lie on a single restriction fragment, here illustrated as 6 kb. With appropriate choice of endonuclease, a given myeloma should always show at least one rearranged band representing the rearranged gene expressed by the myeloma. If the κ allele on the other homologous chromosome has not rearranged, a germ line band may also be visible (blue-green band); alternatively, a second rearranged band may be seen, or there may be no second band at all if the other allele has been deleted.

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
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Tables

Generic image for table
Table 4.1

Biological properties of IgG subclasses

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
Generic image for table
Table 4.2

Properties of Fc receptors

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
Generic image for table
Table 4.3

Some properties of human immunoglobulins a

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
Generic image for table
Table 4.4

Sensitivity of some methods for measuring antibody-antigen reactions a

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
Generic image for table
Table 4.5

A beginner's CD subclassification of lymphoid cells

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4
Generic image for table
Table 4.6

Expression of some CD markers during B-cell differentiation a

Citation: Sell S, Max E. 2001. All about B Cells, p 101-149. In Sell S, Immunology, Immunopathology, and Immunity, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818012.ch4

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