Affinity Maturation of the Primary Response by V Gene Diversification

Immune responses developing in irradiated environment are profoundly altered. The memory anti-arsonate response of A/J mice is dominated by a major clonotype encoded by a single gene segment combination called CRIA. In irradiated and autoreconstituted A/J mice, the level of anti-ARS antibodies upon secondary immunization is normal but devoid of CRIA antibodies. The affinity maturation process and the somatic mutation frequency are reduced. Isotype switching and development of germinal centers (GC) are delayed.The primary antibody response of C57BL/6 mice to the hapten (4-hydroxy-3-nitrophenyl) acetyl (NP)-Keyhole Limpet Hemocyanin (KLH) is dominated by antibodies encoded by a family of closely related VH genes associated with the expression of the λ1 light chain.We investigated the anti-NP primary response in irradiated and autoreconstituted C57BL/6 mice. We observed some splenic alterations as previously described in the irradiated A/J model. Germinal center reaction is delayed although the extrafollicular foci appearance is unchanged. Irradiated C57BL/6 mice are able to mount a primary anti-NP response dominated by λ1 positive antibodies but fail to produce high affinity NP-binding IgGl antibodies. Following a second antigenic challenge, irradiated mice develop enlarged GC and foci. Furthermore, higher affinity NP-binding IgG1 antibodies are detected.

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In Situ Studies of the Primary Immune Response to (4-Hydroxy-3-Nitrophenyl)Acetyl. V. Affinity Maturation Develops in Two Stages of Clonal Selection

Journal of Experimental Medicine ◽

10.1084/jem.187.6.885 ◽

1998 ◽

Vol 187 (6) ◽

pp. 885-895 ◽

Cited By ~ 243

Author(s):

Yoshimasa Takahashi ◽

Pinaki R. Dutta ◽

Douglas M. Cerasoli ◽

Garnett Kelsoe

Keyword(s):

Immune Response ◽

Clonal Selection ◽

Cd40 Ligand ◽

Primary Response ◽

Affinity Maturation ◽

Primary Immune Response ◽

Cellular Interactions ◽

Serum Antibodies ◽

High Affinity ◽

Selection Of

To examine the role of germinal centers (GCs) in the generation and selection of high affinity antibody-forming cells (AFCs), we have analyzed the average affinity of (4-hydroxy-3-nitrophenyl)acetyl (NP)-specific AFCs and serum antibodies both during and after the GC phase of the immune response. In addition, the genetics of NP-binding AFCs were followed to monitor the generation and selection of high affinity AFCs at the clonal level. NP-binding AFCs gradually accumulate in bone marrow (BM) after immunization and BM becomes the predominant locale of specific AFCs in the late primary response. Although the average affinity of NP-specific BM AFCs rapidly increased while GCs were present (GC phase), the affinity of both BM AFCs and serum antibodies continued to increase even after GCs waned (post-GC phase). Affinity maturation in the post-GC phase was also reflected in a shift in the distribution of somatic mutations as well as in the CDR3 sequences of BM AFC antibody heavy chain genes. Disruption of GCs by injection of antibody specific for CD154 (CD40 ligand) decreased the average affinity of subsequent BM AFCs, suggesting that GCs generate the precursors of high affinity BM AFCs; inhibition of CD154-dependent cellular interactions after the GC reaction was complete had no effect on high affinity BM AFCs. Interestingly, limited affinity maturation in the BM AFC compartment still occurs during the late primary response even after treatment with anti-CD154 antibody. Thus, GCs are necessary for the generation of high affinity AFC precursors but are not the only sites for the affinity-driven clonal selection responsible for the maturation of humoral immune responses.

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A multinuclear NMR study of the affinity maturation of anti-NP mouse monoclonal antibodies: Comparison of antibody combining sites of primary response antibody N1G9 and secondary response antibody 3B62

Biochemistry ◽

10.1021/bi00213a028 ◽

1993 ◽

Vol 32 (50) ◽

pp. 13961-13968 ◽

Cited By ~ 6

Author(s):

Tomonori Nakayama ◽

Yoji Arata ◽

Ichio Shimada

Keyword(s):

Monoclonal Antibodies ◽

Primary Response ◽

Affinity Maturation ◽

Secondary Response ◽

Multinuclear Nmr ◽

Nmr Study

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Variant proteins stimulate more IgM+ GC B-cells revealing a mechanism of cross-reactive recognition by antibody memory

eLife ◽

10.7554/elife.26832 ◽

2018 ◽

Vol 7 ◽

Cited By ~ 7

Author(s):

Bronwen R Burton ◽

Richard K Tennant ◽

John Love ◽

Richard W Titball ◽

David C Wraith ◽

...

Keyword(s):

B Cells ◽

Serum Antibody ◽

Primary Response ◽

Affinity Maturation ◽

Memory B Cells ◽

Virus Envelope ◽

B Cell Memory ◽

The Hierarchical Structure ◽

V Genes ◽

Variant Protein

Vaccines induce memory B-cells that provide high affinity secondary antibody responses to identical antigens. Memory B-cells can also re-instigate affinity maturation, but how this happens against antigenic variants is poorly understood despite its potential impact on driving broadly protective immunity against pathogens such as Influenza and Dengue. We immunised mice sequentially with identical or variant Dengue-virus envelope proteins and analysed antibody and germinal-centre (GC) responses. Variant protein boosts induced GCs with a higher proportion of IgM+ B cells. The most variant protein re-stimulated GCs with the highest proportion of IgM+ cells with the most diverse, least mutated V-genes and with a slower but efficient serum antibody response. Recombinant antibodies from GC B-cells showed a higher affinity for the variant antigen than antibodies from a primary response, confirming a memory origin. This reveals a new process of antibody memory, that IgM memory cells with fewer mutations participate in secondary responses to variant antigens, demonstrating how the hierarchical structure of B-cell memory is used and indicating the potential and limits of cross-reactive antibody based immunity.

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Memory in the B–cell compartment: antibody affinity maturation

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2000.0573 ◽

2000 ◽

Vol 355 (1395) ◽

pp. 357-360 ◽

Cited By ~ 46

Author(s):

Michael S. Neuberger ◽

Michael R. Ehrenstein ◽

Cristina Rada ◽

Julian Sale ◽

Facundo D. Batista ◽

...

Keyword(s):

B Cell ◽

Somatic Hypermutation ◽

Primary Response ◽

Affinity Maturation ◽

Immunoglobulin M ◽

Receptor Interaction ◽

Immunoglobulin Gene ◽

Cell Compartment ◽

B Cell Maturation ◽

Genetic Modifications

In the humoral arm of the immune system, the memory response is not only more quickly elicited and of greater magnitude than the primary response, but it is also different in quality. In the recall response to antigen, the antibodies produced are of higher affinity and of different isotype (typically immunoglobulin G rather than immunoglobulin M). This maturation rests on the antigen dependence of B–cell maturation and is effected by programmed genetic modifications of the immunoglobulin gene loci. Here we consider how the B–cell response to antigen depends on the affinity of the antigen–receptor interaction. We also compare and draw parallels between the two processes, which underpin the generation of secondaryresponse antibodies: V gene somatic hypermutation and immunoglobulin heavy–chain class switching.

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Highly mutated memory cells drive an inefficient secondary antibody response to a variant protein

10.1101/438184 ◽

2018 ◽

Cited By ~ 1

Author(s):

Richard K Tennant ◽

Barbara Holzer ◽

John Love ◽

Elma Tchilian ◽

Harry N White

Keyword(s):

B Cells ◽

Antibody Response ◽

Primary Response ◽

Affinity Maturation ◽

Memory B Cells ◽

Secondary Antibody ◽

Cell Compartment ◽

Related Proteins ◽

Serum Response ◽

Variant Protein

AbstractA powerful vaccine against mutable viruses might induce memory antibodies that either strongly bound antigenic variants or that could rapidly undergo secondary affinity maturation to achieve this. We have recently shown after secondary immunization of mice with a widely variant protein (Burton et al. 2018) that IgM+ memory B-cells with few mutations supported an efficient secondary germinal centre (GC) and serum response, superior to a primary response to the same protein. Here, boosting with more closely related proteins produced a GC response dominated by highly mutated B-cells that failed, not efficiently improving serum avidity even in the presence of extra adjuvant, and that was worse than a primary response. This supports a hypothesis that over certain antigenic differences, a cross reactive, mutated, memory B-cell compartment can be an impediment to affinity maturation.

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Possible contribution of DNase γ to immunoglobulin V gene diversification

Immunology Letters ◽

10.1016/j.imlet.2009.05.005 ◽

2009 ◽

Vol 125 (1) ◽

pp. 22-30 ◽

Cited By ~ 3

Author(s):

Noriaki Okamoto ◽

Mariko Okamoto ◽

Shinsuke Araki ◽

Hiroshi Arakawa ◽

Ryushin Mizuta ◽

...

Keyword(s):

Dnase Γ ◽

Gene Diversification ◽

V Gene

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Immunoglobulin D (IgD)-deficient mice reveal an auxiliary receptor function for IgD in antigen-mediated recruitment of B cells.

Journal of Experimental Medicine ◽

10.1084/jem.177.1.45 ◽

1993 ◽

Vol 177 (1) ◽

pp. 45-55 ◽

Cited By ~ 161

Author(s):

J Roes ◽

K Rajewsky

Keyword(s):

T Cell ◽

B Cells ◽

B Cell ◽

Primary Response ◽

Affinity Maturation ◽

Resting Cells ◽

Wild Type ◽

Normal Numbers ◽

Immunoglobulin D ◽

Deficient Mice

To assess the role of immunoglobulin D (IgD) in vivo we generated IgD-deficient mice by gene targeting and studied B cell development and function in the absence of IgD expression. In the mutant animals, conventional and CD5-positive (B1) B cells are present in normal numbers, and the expression of the surface markers CD22 and CD23 in the compartment of conventional B cells indicates acquisition of a mature phenotype. As in wild-type animals, most of the peripheral B cells are resting cells. The IgD-deficient mice respond well to T cell-independent and -dependent antigens. However, in heterozygous mutant animals, B cells expressing the wild type IgH locus are overrepresented in the peripheral B cell pool, and T cell-dependent IgG1 responses are further dominated by B cells expressing the wild-type allele. Similarly, in homozygous mutant (IgD-deficient) animals, affinity maturation is delayed in the early primary response compared to control animals, although the mutants are capable of generating high affinity B cell memory. Thus, rather than being involved in major regulatory processes as had been suggested, IgD seems to function as an antigen receptor optimized for efficient recruitment of B cells into antigen-driven responses. The IgD-mediated acceleration of affinity maturation in the early phase of the T cell-dependent primary response may confer to the animal a critical advantage in the defense against pathogens.

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