Mechanism of formation of univalent fragments of rabbit antibody

Nisonoff, Alfred; Wissler, F.C.; Woernley, D.L.

doi:10.1016/0006-291x(59)90046-4

Cited by 55 publications

(15 citation statements)

References 4 publications

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“…Of particular interest in these studies is the demonstration of the existence of at least two distinct antigenic fragments. While comparison of these with native y-globulin suggests neither loss nor release of antigenic groups present in the native molecule, preliminary studies, comparing them to fragments produced by treatment with pepsin and mercaptoethanol as described by Nisonoff and associates (8), suggest the possible loss of a few antigenic groups present in the native protein following digestion with papain.…”

Section: Discussionmentioning

confidence: 82%

“…However, the observations that a number of closely related smaller fragments can be prepared from rabbit and human y-globulins by sulfhydryl reagents combined with denaturing agents (30), papain (4,5) or pepsin (8) suggest that the common mechanism may be the reduction of disulfide bonds by the sulfhydryl reagent, and that the enzyme or denaturing agent may make these accessible by unfolding the molecule or by breaking off a fragment. While the close similarity of the antigenic properties of fractions A and C when tested with rabbit antisera raises the possibility that they are identical, such a view does not appear likely, since they differ in some of their chemical properties, in electrophoretic mobility, and in antigenicity when tested with antisera produced in different species (31 units.…”

Section: Discussionmentioning

confidence: 99%

“…The exact nature of these products was not examined further. In a number of experiments, 3 mg pepsin per 100 mg 'y-globulin at pH 4 was used, alone or in conjuunction with subsequent treatment, with 0.1 M mercaptoethanol as described by Nisonoff, Wissler and Woernley (8).…”

Section: Methodsmentioning

confidence: 99%

“…More recently Porter (4,5) has produced split products of rabbit y-globulin by treatment with papain and cysteine, two of which retained antibody activity as measured by their ability to inhibit precipitation of the original antiserum with homologous antigen, while the third appeared to retain the greatest antigenic specificity. Karush (6) and Nisonoff, Woernley and Wissler (7,8) working with antihapten antibodies have clearly shown that these fragments are univalent. Putnam, Hsiao and Tan (9), employing similar techniques, have been able to split human y-globulin and myeloma proteins into two major chromatographically separable fractions with sedimentation coefficients of approximately 3.5S.…”

mentioning

confidence: 99%

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Structural Units of Human 7s Gamma Globulin *

Franklin¹,

Prelli²

1960

J. Clin. Invest.

184

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Section: Discussionmentioning

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Structural Units of Human 7s Gamma Globulin *

Franklin¹,

Prelli²

1960

J. Clin. Invest.

184

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“…The globulins so precipitated were dissolved in 5 ml of saline and dialyzed against BBS in the cold and the IgG2 fraction was isolated by preparative agar block electrophoresis in barbital buffer (pH 8.6,/im-0.1), as described in a previous study (7). The F(ab')2 fragments were then prepared by the method of Nisonoff (15) with slight modifications. Briefly, 20 mg of IgG2 were incubated with 0.6 mg of pepsin (Sigma Chemical Co., St. Louis, No.)…”

Section: Preparation Of Antiseramentioning

confidence: 99%

Mechanisms of regulation of cell-mediated immunity. III. The characterization of azobenzenearsonate-specific suppressor T-cell-derived-suppressor factors.

Greene¹,

Bach²,

Benacerraf³

1979

The Journal of Experimental Medicine

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Delayed type hypersensitivity to the hapten azobenzenearsonate (ABA) can be induced and suppressed by the administration of hapten-coupled syngeneic spleen cells by the appropriate route. Suppressor T cells stimulated by the intravenous administration of ABA-coupled spleen cells have been shown to produce a discrete subcellular factor(s) which is capable of suppressing delayed type hypersensitivity to azobenzenearsonate in the mouse. Such suppressor factors may be produced by the mechanical disruption of suppressor cells or by placing such suppressor cells in culture for 24 h. The suppressor factor(s) (SF) derived from ABA-specific suppressor cells exhibit biological specificity for the suppression of ABA delayed type hypersensitivity (DTH), but not trinitro-phenyl DTH, as well as the capacity to bind to ABA immunoadsorbents. Passage of suppressor factor(s) over reverse immunoadsorbents utilizing a rabbit anti-mouse F(ab')2 antiserum demonstrated that the antigen-specific T-cell derived SF does not bear conventional immunoglobulin markers. The suppressor factor(s) are not immunoglobulin molecules was further demonstrated by the inability of anti-ABA antibodies to suppress ABA DTH. Gel filtration of ABA suppressor factor(s) showed that the majority of the suppressive activity was present in a fraction with molecular weight ranging between 6.8 x 10(4) and 3.3 x 10(4) daltons. We also analyzed for the presence of determinants encoded by the H-2 major histocompatibility complex (MHC) and found that immunoadsorbents prepared utilizing antisera capable of interacting with gene products of the whole or selected gene regions of H-2 MHC, i.e., B10.D2 anti-B10.A and B10 anti-B10.A immunoadsorbents, retained the suppressive activity of ABA-SF. Elution of such columns with glycine HCl buffers (pH 2.8) permitted recovery of specific suppressive activity. Taken collectively such data supports the notion that suppressor T-cell-derived ABA suppressor factors have antigen-binding specificity as well as determinants controlled by the K end of the H-2 MHC. The distribution of strains capable of making SF has also been analyzed. The relationship of the antigen-binding specificity to VH gene products is discussed in this and the companion paper.

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Human thymus antigen: Characterization and its expression on human leukemias

et al. 1981

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Specific anti-human thymus xenoantiserum (ATS) was utilized for characterizing a human thymus antigen (HTA) preferentially expressed on human thymocytes. Binding of ATS with different cell types was studied by immunofluorescence and immunoperoxidase techniques, as well as by radioimmunoprecipitation (RIP) followed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). ATS reacted selectively with the majority (87%) of human thymocytes among normal lymphoid cell populations tested. In the thymus, HTA was expressed largely on cortical thymocytes but not on those cells located in the thymic medulla. When 20 cases of different types of lymphatic leukemias were studied with ATS, HTA was found on acute lymphoblastic leukemia cells rosetting with sheep erythrocytes (T-ALL) but not on other lymphatic leukemia cases, including chronic lymphocytic leukemia cells rosetting with sheep red blood cells (T-CLL). SDS-PAGE analysis of immunoprecipitates formed between ATS and radiolabeled cell-surface components of human thymocytes demonstrated that ATS bound a 48,000-molecular-weight component that could be adsorbed to Sepharose 4B coupled with Les culinaris hemagglutinin and could be labeled by periodate-tritiated borohydride, indicating that HTA is a sialoglycoprotein. Similar antigen molecules were also precipitated by ATS from T-ALL cells but not from other lymphatic leukemia cells. Five T-ALL-derived cell lines were tested with ATS by immunofluorescence and by RIP and SDS-PAGE, but only two cell lines (MOLT-3 and P12/Ich) possessed HTA molecules on their cell membrane.

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Mechanism of formation of univalent fragments of rabbit antibody

Cited by 55 publications

References 4 publications

Structural Units of Human 7s Gamma Globulin *

Structural Units of Human 7s Gamma Globulin *

Mechanisms of regulation of cell-mediated immunity. III. The characterization of azobenzenearsonate-specific suppressor T-cell-derived-suppressor factors.

Human thymus antigen: Characterization and its expression on human leukemias

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