The molecular evolution of the immune response

Manser, Tim; Wysocki, Lawrence J.; Gridley, Thomas; Near, Richard I.; Gefter, Malcolm L.

doi:10.1016/0167-5699(85)90024-6

Cited by 98 publications

(53 citation statements)

References 40 publications

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“…VHIdcRexpressing hybridomas isolated at later times (days 10-21) during the primary Ars-KLH response (Manser, T., unpublished results) and during the secondary response (1) are characterized by restricted gene segment combinational diversity (a V domain encoded by a single combination of gene segments termed the "canonical" combination predominates), extensive somatic mutational alteration, and increased (as compared with early primary antibodies) affinity for Ars (Ka from 105 to 10'/M) . We previously suggested that an antigen-driven process of V gene somatic mutation and clonal selection of B cells expressing high-affinity surface Ig can best explain this maturation of the anti-Ars-KLH response (36) .…”

Section: Resultsmentioning

confidence: 99%

Influence of the macromolecular form of a B cell epitope on the expression of antibody variable and constant region structure.

Fish¹,

Manser²

1987

The Journal of Experimental Medicine

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We investigated the influence of the macromolecular form of an epitope on the structure of antibody variable and constant regions expressed by the B cell population participating in an immune response to that epitope. Hybridomas were constructed from strain A/J mice undergoing either primary or secondary immune responses to p-azophenylarsonate conjugated to Brucella abortus (Ars-Bruc). We determined the sequences of the V genes expressed by hybridomas selected on the basis of expression of a single VH gene segment known to encode a large family of anti-Ars antibodies. These sequences were compared with the sequences of V genes expressed by a previously characterized panel of hybridomas isolated in the same way during the primary and secondary responses of A/J mice to Ars-KLH. The repertoire of Ars-specific V domains expressed among primary and secondary hybridomas elicited with these two forms of Ars were similar, as were the differences between primary and secondary V region somatic mutational alteration and affinity for Ars. In contrast, predominant expression of IgG2 anti-Ars antibodies was elicited in the secondary Ars-Bruc response, whereas secondary anti-Ars antibodies elicited with Ars-KLH are predominantly IgG1. Thus, differences in the macromolecular form of Ars clearly influence the isotypic profile of the anti-Ars response, but the expression, diversification, and selection of V domains elicited with this hapten are not greatly affected by such differences. Our results suggest that while isotype regulation is highly perceptive of the macromolecular form of a B cell epitope, V region regulation is primarily influenced by the molecular structure of that epitope.

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Section: Resultsmentioning

confidence: 99%

Influence of the macromolecular form of a B cell epitope on the expression of antibody variable and constant region structure.

Fish¹,

Manser²

1987

The Journal of Experimental Medicine

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“…While this generalization has been to a large extent developed from the study of VH gene segments (1)(2)(3)(4)(5)(6), the situation is likely the same for light chains although fewer systems have been systematically studied (7,8). Several mechanisms may contribute to generate diversity in the secondary immune response (9)(10)(11)(12)(13)(14).…”

mentioning

confidence: 99%

V kappa and J kappa gene segments of A/J Ars-A antibodies: somatic recombination generates the essential arginine at the junction of the variable and joining regions.

Sanz¹,

Capra²

1987

Proc. Natl. Acad. Sci. U.S.A.

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The germ-line elements for the variable (V) and joining (J) regions of the K chains (VK.ArS, Jh1) that give rise to the productive allele for the A/J mouse light chains of Ars-A monoclonal antibodies (directed against the haptenp-azophenylarsonate) have been cloned and sequenced as well as a rearranged Ars-A light chain gene. Using the VK ,A, gene as a hybridization probe, we provide evidence that the VKo1 family is relatively small and that only one member gives rise to the productive allele in all Ars-A antibodies. An unusual feature of Ars-A light chains is that all contain an arginine at position 96, the V-J junctional position. We had shown by chain recombination that arginine-96 was essential for antigen binding. In the present study, an arginine codon was not found either at the 3' end of the V,,^Ars gene segment or at the 5' end ofJK1. However, an arginine codon (CGG) can easily be generated by recombination between these two germ-line elements. Thus, we document that junctional diversity through intracodonic recombination can be crucial to the antibody binding function of the resulting molecule. In the majority of primary immune responses to different haptens so far studied in detail, the antibodies may be subdivided into a small number offamilies, each of which can usually be shown to result from the somatic diversification of a few germ-line heavy-and/or light-chain variable-region (VH and VL, respectively) gene segments. While this generalization has been to a large extent developed from the study of VH gene segments (1-6), the situation is likely the same for light chains although fewer systems have been systematically studied (7,8). Several mechanisms may contribute to generate diversity in the secondary immune response (9-14).In the A/J strain of mice, the humoral immune response to the hapten p-azophenylarsonate (Ars) coupled to keyhole limpet hemocyanin may be subdivided serologically and by amino acid sequence criteria into three families of monoclonal antibodies termed Ars A, Ars B, and Ars C (15, 16). The Ars-A family comprises all the strain-specific crossreactive idiotype-(CRI) positive molecules (17) and also the CRInegative molecule 91A3 whose light chain has been shown by chain-recombination experiments to be CRI positive in nature (18,19).To date, 10 light chains have been completely sequenced either at the protein (16)(17)(18) or mRNA (20) level from Ars-A hybridomas. With the exception of 91A3, all were derived from CRI-positive molecules. From these data a consensus sequence was generated, and the sequenced Ars-A light chains differ from this consensus by from 1 to 5 amino acids (21). Since the pattern of these substitutions appears to be nearly random, we postulated that all of these expressed genes derive from the same germ-line elements. The 91A3 light chain, the subject of this paper, is the most distant from the consensus sequence, differing in 5 of the 108 positions in its variable (V) region (reviewed in ref. 21).A striking characteristic of Ars-A light chains is ...

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“…A droplet (20 Al) of the protein/PEG solution was equilibrated against a larger volume (typically, 1 ml) of 8.5% PEG 8000/50 mM potassium phosphate, pH 7.5/0.5% NaN3 at room temperature in a sealed dish. Crystals appeared after [5][6][7] …”

mentioning

confidence: 99%

Crystal structure of the antigen-binding fragment of the murine anti-arsonate monoclonal antibody 36-71 at 2.9-A resolution.

Rose

Strong

Margolies

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

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The structure of the antigen-binding fragment (Fab) of an anti-phenylarsonate monoclonal antibody (36-71) bearing a major crossreacting idiotype of A/J mice has been solved and refined to an R factor of 19.3% at a resolution of 2.9 A. An initial electron density map was obtained with phase information from a total of six isomorphous heavy-atom derivatives (from two different compounds) and a molecular replacement solution using the HED10 Fab crystal structure as a model. The structure of the McPC603 Fab was used to provide an initial set of atomic coordinates. The electron density maps are clear and easily interpretable for the entire sequence except for sections from two of the heavy-chain complementarity-determining regions totaling 21 residues.These residues have been left out of the refinement and are not represented in our current model. The antigen-combining site was located by means of a difference Fourier synthesis with one of the heavy-atom derivatives, which contained arsanilic acid. It lies in a small pocket formed by residues from the hypervariable regions of both the heavy and the light chains. Interactions with the hapten from framework residues are also possible.Monoclonal antibodies specific to the hapten p-azophenylarsonate are proving to be useful probes into the structural basis of antigen-antibody and anti-idiotype-idiotype interactions. When A/J strain mice are immunized with this hapten conjugated to a keyhole limpet hemocyanin carrier, the majority of the antibodies produced in response are members of a crossreacting idiotype family (CRIA or IdCR+) (1, 2). The evidence to date indicates that all IdCR+ anti-arsonate antibodies are the result, in the secondary immune response, of the combination of a single set of gene segments encoding variable (V) regions of immunoglobulins (3). Secondary immune-response anti-arsonate antibodies bearing IdCR typically contain somatic mutations that, on average, result in antibodies with higher affinity for the hapten (reviewed in ref. 4). The dominance of IdCR was explained by a relatively high intrinsic affinity of the unique germ-line-encoded V domain expressing IdCR and the ability of that domain to sustain somatic mutations that enhance affinity (5). Alternative explanations of idiotype dominance involve the regulation of antibody production by interactions ("network") of idiotypes with anti-idiotypes (6). The structural features of antiarsonate antibodies that account for hapten specificity and their idiotypic determinants have thus been a subject for intense scrutiny (reviewed in refs. 4 and 7).We have undertaken to crystallize the Fab fragments of some of these antibodies to provide a three-dimensional structural basis for the interpretation of the sequence information. We succeeded in growing crystals of the Fab fragments of two IdCR+ antibodies: 36-65 and 36-71 (both IgG1, K). The most suitable of these two crystal forms, that of the 36-71 Fab, diffracts beyond 2.5-A resolution and is well behaved in x-ray experiments; the three-dimensiona...

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The molecular evolution of the immune response

Cited by 98 publications

References 40 publications

Influence of the macromolecular form of a B cell epitope on the expression of antibody variable and constant region structure.

Influence of the macromolecular form of a B cell epitope on the expression of antibody variable and constant region structure.

V kappa and J kappa gene segments of A/J Ars-A antibodies: somatic recombination generates the essential arginine at the junction of the variable and joining regions.

Crystal structure of the antigen-binding fragment of the murine anti-arsonate monoclonal antibody 36-71 at 2.9-A resolution.

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