Plaque Formation in Agar by Single Antibody-Producing Cells

Gilden, Raymond V.; Tokuda, Sei

doi:10.1126/science.140.3565.405-a

Cited by 19 publications

(14 citation statements)

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“…Our results are consistent with earlier findings that after repeated antigenic stimulation antlsera form unusually stable complexes with the corresponding antigens (5)(6)(7)(8)(9). Although a number of nonspecific factors may influence the interaction of antibodies with antigens, the stability of these complexes is probably largely the result of an increase in the affinity of the antibody-binding sites for the various antigenic determinants.…”

Section: Discussionsupporting

confidence: 92%

“…For example, several weeks (or even years) after the first injection, a second exposure to the immunogen often evokes an unusually vigorous response: the serum antibody concentration typically increases rapidly to reach and remain at a level higher than that attained after the primary stimulus (1)(2)(3)(4). In addition, the antibodies produced in the secondary response often form more stable complexes with antigen than those formed at a comparable time after the first injection (5)(6)(7)(8)(9).…”

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confidence: 99%

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The Relative Affinity of Antibodies Synthesized in the Secondary Response

Steiner

Eisen

1967

The Journal of Experimental Medicine

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The immune response to repeated antigenic stimtflation differs in a number of respects from the response to the initial exposure to immunogen. For example, several weeks (or even years) after the first injection, a second exposure to the immunogen often evokes an unusually vigorous response: the serum antibody concentration typically increases rapidly to reach and remain at a level higher than that attained after the primary stimulus (1-4). In addition, the antibodies produced in the secondary response often form more stable complexes with antigen than those formed at a comparable time after the first injection (5-9).Recent studies of antibodies isolated from serum at various intervals after immunization with 2,4-dinitrophenylated proteins have shown that the affinity of these antibodies for simple dinitrophenyl haptens (expressed as the average intrinsic association constant) is greater the longer the period between immunization and bleeding, and the smaller the dose of immunogen (10). These properties of the serum antibodies are the direct result of a sequential change in the nature of the antibodies synthesized by lymph node cells (11). It seemed possible, therefore, that the affinity of the antibodies formed at any time after a single injection of antigen is inversely related to the amount of antigen remaining in the animal. Accordingly, we wished to determine the effect on antibody affinity of a second injection of antigen, given when the animal is already forming high affinity antibody in response to the initial stimulus.In experiments reported here, it is demonstrated that the typical response to a second injection of antigen is a burst of synthesis of antibodies with much greater a /~i t y for the dinitrophenyl group than those formed at a comparable time after the primary immunization with the same dose of immunogen. Indeed, the earliest antibodies formed after restimulation are already high in affinity.

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Section: Discussionsupporting

confidence: 92%

mentioning

confidence: 99%

The Relative Affinity of Antibodies Synthesized in the Secondary Response

Steiner

Eisen

1967

The Journal of Experimental Medicine

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“…Two explanations for such an event are at hand: (a) two antigens share common determinants which are recognized by the immunized animal, and (b) cross-reactivity is a function of cross-stimulation of two antibody populations in the antiserum, one specific for one and the other for the related antigen. This is in essence the phenomenon of the anamnestic response as it was observed for various strains of influenza viruses (1-3), the DNP and TNP ligands (4), sulfanilate-metanilate haptenic groups (5), and some serum proteins (8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20).…”

Section: Figmentioning

confidence: 93%

Cross-Stimulation of Monoclonal Antibodies in Anamnestic Responses

Cramer¹,

Braun²

1973

The Journal of Experimental Medicine

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Immunization of BALB/c mice with Group A streptococcal vaccines leads to the induction of high levels of monoclonal antibody populations. Subsequent immunization of these mice with Group A-variant streptococcal vaccines induces a significant level of monoclonal antibody of A-variant antigen specificity revealed by labeled group A-variant polysaccharide. During these course Av and Av' immunizations, the monoclonal Group A-specific antibodies were also restimulated to levels usually higher than the variant-specific antibodies. With two exceptions, these homogeneous antibody populations were not cross-reactive in vitro with the related antigen. Such cross-stimulation of monoclonal antibodies was interpreted as a function of particular membrane properties of the Ig receptor-bearing memory cells which, for restimulation, would only require the structurally closely related antigen which serves as a backbone to the original antigen, and not necessarily the exact fit of the homologous immunodominant group.

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“…This can be explained by the fact that, even after repeated immunization, only a portion of an animal's B-cell repertoire becomes reactive to the antigen; a larger portion remains unreactive and presumably naïve (Story et al, 2008). Thus, the ability of the immune system of a hyperimmunized animal to elicit humoral response to a second antigen (Gilden and Tokuda, 1963;Pleten, 2007) could be reproduced in the phage display system. These results demonstrate that a sufficiently large immune library can be a ready source of antibodies, not only against the immunizing antigen but also against many other non-immunizing antigens.…”

Section: Resultsmentioning

confidence: 99%