Production of auto-anti-idiotypic antibody during the normal immune response to TNP-ficoll. II. Hapten-reversible inhibition of anti-TNP plaque-forming cells by immune serum as an assay for auto-anti-idiotypic antibody

Abstract

In the accompanying paper (1), we have shown that after day 4 of the immune response of AKR/J mice to 2,4,6-trinitrophenyl-lys-Ficoll (TNP-F) 1, the addition of free hapten to a plaque-forming-cell (PFC) assay increased the number of observed splenic anti-trinitrophenol (TNP) PFC. Immune spleen cells, taken 7 d after immunization, transferred this property of the immune response to normal recipients; spleen cells from such recipients, assayed 4 d after cell transfer and TNP-F injection, manifested an exaggerated form of this phenomenon. It was hypothesized that the increase in PFC was the result of the displacement, by hapten, of auto-anti-idiotypic antibodies that were synthesized during the course of the normal immune response. If this hypothesis were correct, it should be possible to obtain auto-anti-idiotypic antibodies by hapten elution from appropriate immune spleen cells. Results in the accompanying paper (1) also suggested that the putative auto-anti-idiotypic-antibody response was involved in the downward regulation of the immune response of AKR/ J mice to TNP-F. One might, therefore, expect to find auto-anti-idiotypic antibody in the serum of AKR/J mice immediately after the abrupt decrease in the number of detectable splenic PFC: i.e., 7 d after antigen injection.In the present paper, evidence is presented to support these hypotheses in that hapten-reversible inhibition of PFC in vitro can be demonstrated with hapten eluates from immune cells and with immune serum. The factor responsible for inhibition has immunoglobulin-like determinants, lacks anti-TNP-antibody activity, and is absorbable by an AKR/J anti-TNP-antibody immunoadsorbent. Hapten-reversible inhibition of PFC represents a simple in vitro assay for anti-idiotypic antibody. In this

Strain Differences in Production of Direct Plaque‐Forming Cells in Response to DNP‐Ficoll, a Thymus‐Independent Antigen, in Mice

“…The distinctive characteristics of this response to the thymus-independent antigens in mice are the restriction of the antibody response to IgM and IgG 2 classes (13), the response by ontogenetically late-appearing B cells lacking complement receptors (6) and expressed surface IgD receptors (9), the discovery of X-linked unresponsiveness in an inbred strain of mice (l), and the regulation of antibody production by anti-idiotypic antibodies in vivo (3,12).…”

“…The distinctive characteristics of this response to the thymus-independent antigens in mice are the restriction of the antibody response to IgM and IgG 2 classes (13), the response by ontogenetically late-appearing B cells lacking complement receptors (6) and expressed surface IgD receptors (9), the discovery of X-linked unresponsiveness in an inbred strain of mice (l), and the regulation of antibody production by anti-idiotypic antibodies in vivo (3,12).In the course of analyzing the response of various strains of mice to DNPFicoll, we realized that anti-DNP or anti-TNP plaque-forming cell (PFC) response is strain specific. However, these strain differences were observed only in in vivo experiments; they were not demonstrable in vitro.…”

Cellular interactions in immune regulation. Hapten-specific suppression by non-T cells and T cell-mediated reversal of suppression

“…The anti-idiotype antibody might bind the receptors of the assay B cells and inhibit antibody secretion. Precedents exist in the literature for both mechanisms (1,2,18). A soluble specific suppressor factor was detected in the supernate from immune spleen cells cultured in the absence of antigen.…”

Age-Related Strain Differences in the Development of Auto-Anti-Idiotypic Antibody Regulation in the Splenic and Mucosal-Associated Lymphoid Systems

“…Other studies [11] have described an assay for anti-idiotypic antibody in immune sera, the basis of this assay being the hapten-(TNP-EACA-)reversible inhibition of idiotype-producing PFC. Results in table III show that pools of immune sera from high hapten-angmentable PFC-producing mice (previously immunized with TNP-BGG in CFA 14 days before) caused a hapten-revers ible block of plaque formation in spleen cells from either TNP-dextran or TNP-BGG im mune C57BL/6J mice.…”

“…This suppressive effect of auto-anti-idiotypic antibody was at tributed possibly to its simultaneous binding of surface Ig and Fc receptors on the surface of the B cell, thus inhibiting antibody secre tion [11], In previous studies, we have shown that the decline in the number of plaque forming cells (PFC), loss of high affinity PFC, and the increase in hapten-augmentable PFC with age in the spleen of C57BL/6J male mice may be due to auto-anti-idiotypic anti body regulation [28], In contrast, mucosal immunity in old C57BL/6J mice, as mea sured in PFC responses of the mediastinal (BLN) and mesenteric (MLN) lymph nodes, was not reduced [31]; further, there was no loss of high affinity antibody-secreting cells [30], and there was no appreciable appear ance of anti-idiotype-blocked, hapten-aug mentable PFC [29] in these areas. These find ings support the hypothesis that the systemic and mucosal B cell repertoires do not un dergo parallel changes with age [33].…”