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M Feldmann

Publications and source records attributed to M Feldmann.

At least 361 records · Page 20Linked to original sources

Nature of macrophage-T cell interaction in secondary helper cell generation in vitro. Genetic restriction of macrophage-T cell interaction, which determines T-B genetic restriction.

To investigate the histocompatibility requirements for the macrophage-T cell interaction in the secondary antibody response, splenic T cells from antigen (carrier)-primed F1 hybrid mice were restimulated in vitro with carrier-pulsed F1, parental or allogeneic macrophages. Surviving T cell were cocultured with hapten-primed F1 or parental "B cells" and restimulated with the appropriate hapten-carrier conjugate. The IgG antibody-forming cell response was then measured using a plaque assay. Mapping of the genetic restriction was performed by use of different strain combinations. T helper cells could be restimulated in the presence of macrophages only provided they shared the I-A subregion of the major histocompatibility complex with the F1 T cells frm F1 hybrids restimulated with parental or I-A-identical macrophages were shown to only cooperate with parental B cells of the same I-A haplotype as the macrophages used for restimulation. The defect was at the level of the macrophage, as addition of macrophages of the I-A haplotype used for the restimulation culture reconstituted ability of F1 helper cells to cooperate with the I-A-nonidentical B cells.

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Antigen-specific helper factor reacts with antibodies to human beta 2 microglobulin.

Antigen-specific helper factor was induced in vitro from lymphoid cells of monkeys and mice by using an antigen derived from Streptococcus mutans. Helper activity was removed from supernatants of monkey cells by affinity chromatography on Sepharose 4B insolubilized antibodies specific for human beta 2-microglobulin (H beta 2M) prepared in chicken, rabbit and rat, and an insolubilized monoclonal mouse anti-H beta 2M antibody-bound monkey helper factor activity. However, guinea pig antibody to human beta 2M was inactive. In parallel studies, the pattern of absorption of mouse helper factor (HF) was different from that of the monkey in that insolubilized guinea pig anti-H beta 2M bound helper factor, whereas rabbit and monoclonal anti-H beta 2M failed to do so. Although these findings were not compatible with an intact beta 2M chain being present in helper factor, they may imply a cross-reactivity of beta 2M with a "constant region" of helper factor that may share common sequences with beta 2M. This may suggest that factor genes have evolved from the same ancestral genes as beta 2M.

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T-cell hybridomas producing hapten-specific suppressor factors.

We have made several T-T hybridomas which secrete soluble factors capable of suppressing an in vitro antibody response to nitrophenol (NP), but not other unrelated antigens. These factors bind specifically to NP, and express determinants coded for in the I-J region of the mouse major histocompatibility complex. No determinants that cross-react with the constant regions of mouse immunoglobulins are present on the factors. Three sub-clones originating from the same initial culture well of hybridoma cells secrete factors which carry I-J determinants of different haplotypes. One clone expresses I-J determinants derived from the suppressor cell parent, another expresses I-J determinants derived from the tumour cell parent, and a third expresses both. This correlates exactly with I-J determinants expressed on the cell membrane, and suggests the participation of at least two genes in the determination of suppressor-factor structure.

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Induction of human antigen-specific suppressor factors in vitro.

Based on methods used for the in vitro induction of antigen-specific suppressor cells in the mouse, we have cultured Ficoll-Isopaque-separated human blood cells with high dose of antigen (100 microgram/ml) in Marbrook culture vessels for 4 days. The resulting cells, when further recultured for 24 hr with a low dose of antigen (1 microgram/ml), released into the supernatant material, termed 'suppressor factor', which inhibited, in an antigen-specific manner, the antibody response of mouse spleen cells in culture. The suppressor factor was analysed using immunoabsorbents, and was bound to and eluted from specific antigen, concanavalin A and lentil lectin, anti-human Ia antibodies, and anti-mouse suppressor factor antibodies, but was not bound to antibodies against human IgG.

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Genetics of human T cell-monocyte interaction in helper cell induction.

The generation of human antigen-specific helper cells from unprimed peripheral blood lymphoid cells in tissue culture requires the presence of the appropriate number of blood monocytes. The role of the HLA complex in this cell interaction was investigated, by using HLA-type donors, and it was found that the monocytes had to share at least one HLA-DR specificity with the T cell donor. These experiments suggested that the genes controlling the macrophage-T cell interaction are closely associated or in linkage disequilibrium with the HLA-DR region.

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Dendritic cells induce T cell proliferation to synthetic antigens under Ir gene control.

Dendritic cells prepared by a modification of the method of Steinman and Cohn are I-A+ and FcR-. They are extremely potent at activating not only allogeneic T cell proliferation but also antigen-specific syngeneic T cell proliferation. Dendritic cells from nonresponder strains are unable to present antigens to responder X nonresponder T cells, suggesting that they may be a site of Ir gene product expression.

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Antigen specific T cell factors.

Antigen specific helper and suppressor factors have a similar structure, with two major sections, a 'variable region', determining antigen specificity which is likely to be controlled by Immunoglobulin VH genes, with which it shares idiotype and framework determinants. Specific factors also have a 'constant region' which does not vary between strains and minimally between species or with the antigenic specificity of the factors, which are defined by rabbit anti-helper or anti-suppressor antisera. This region determines the biological function of the molecule. Anti-Ia antisera react with factors, but the nature and function of Ia molecules on T cell factors is still unclear. The model of specific factor structure, with C and V regions resembles that of immunoglobulin, and it is thus possible that the C region of factors, like the V region is Ig linked. Because there are multiple T cells, helping and suppressing antibody responses specifically, it seems improbable that all of these cells could interact directly with rare antigen-specific B cells. Thus we propose that macrophage presenting cells are the key to the integration of signals for immune induction and regulation for T and B cells. Since Ir genes have been identified in the macrophage presenting cells interacting with both T and B cells, this suggests that macrophage Ia antigens are of importance in the integration of triggering signals for the lymphoid pool.

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Nature of T-cell macrophage interaction in helper-cell induction in vitro. II. Two stages of T-helper-cell differentiation analyzed in irradiation and allophenic chimeras.

The genetic restriction in the T-cell-macrophage-like cell interaction in helper cell induction was investigated with allophenic and irradiation chimeras of various types. Using T cells from P leads to F1 chimeras, there was a restriction of cooperation with the parental haplotype accessory cells, unless the chimeric mice were repopulated with macrophages of the opposite haplotype before priming. T cells from primed or unprimed F1 leads to P chimeras only cooperated with recipient type accessory cells. These observations led to the hypothesis that there are two stages in the genesis of immunocompetence of T helper cells, one dependent on the thymus, and the other on peripheral macrophage-like cells. Purified T cells from P1 + P2 leads to F1 irradiation chimeras behaved in an unexpected manner in the unprimed state, preferring to cooperate with their own haplotype macrophages. This self preference was lost after antigen priming in vivo and was not noted in allophenic chimeras. This loss of self preference was restricted to the haplotypes represented in the chimeras, and did not extend to third party haplotypes. While these in vitro induced helper cells from chimeric mice show clear genetic restrictions at the T-cell macrophage-like cell interaction, there was no evidence for a matching T-B genetic restriction.

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Serological analysis of antigen-specific helper factors specific for poly-L(Tyr, Glu)-poly-DLAla--poly-LLys [(T, G)-A--L] and L Glu60-LAla30-LTyr10 (GAT).

In vitro prepared antigen-specific helper factors reactive to the synthetic polypeptide antigens poly-L(Tyr, Glu)-poly-DLAla--poly-LLys [(T, G)-A--L] or LGlu60-LAla30-LTyr10 (GAT) and bearing Ia determinants were analyzed serologically to determine the nature of the Ia determinants they expressed. I subregion-specific mouse anti-Ia antisera were used, and showed that (T, G)-A--L-specific helper factor (HF) contains I-A subregion-controlled determinants, whereas GAT-specific HF carries I-J subregion-controlled antigens. This unexptected finding was confirmed in both the H-2k and H-2 b haplotypes, using a variety of anti-I-J antisera. Rabbit anti-Ia antisera also reacted with both HF which raised the possibility that the Ia determinants on HF may be carbohydrate in nature. The fact that HF has a low molecular weight and yet contains Ia determinants, antigen-binding capacity and idiotypic markers is compatible with this interpretation.

Adsorption↗

Comparison of antigen-specific I-region-associated cell interaction factors.

Two basic types of factors reacting with anti-I region (anti-Ia) antisera are compared, those derived from macrophage-like antigen presenting cells and others derived from T-lymphocytes, of either the suppressor or helper type. Despite the common property of reacting with anti-Ia antisera, the two sets of factors differ by many criteria. Macrophages, upon culture with antigen, release complexes of Ia antigen and a fragment of the original immunogen. This material is only produced by responder macrophages and thus appears to be a soluble Ir gene product. The genetic restriction of the T-macrophage interaction was investigated in chimeras, and it was found that the host environment as well as the donor genotype was of importance in determining restrictions, which were thus not really directed to "self." There was no evidence for intrinsic T-cell Ir genes, as nonresponder stem cells developed into responder T-cells in a (responder X nonresponder) F1 environment. However, these cells only responded in the presence of responder macrophages. Specific T-cell factors are different in nature. These all react with anti-Ia antisera, but the nature or function of the T-cell Ia is unknown. The basic structure involves a VARIAble region" responsible for antigen binding which, as it reacts with anti-idiotype antisera and anti-variable region framework antisera is an immunoglobulin variable region. There is also a "constant region," defined by its biological properties as well as by specific rabbit antisera. This two-region nature of specific factors is reminiscent of immunoglobulin structure and it is a reasonable hypothesis that the constant region is linked to the Ig cluster of genes.

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Is genetically related macrophage factor (GRF) a soluble immune response (Ir) gene product?

The possibility that the antigen-presenting "macrophages" interacting with helper cells either directly or via the intermediary action of a soluble factor consisting of Ia antigen and a fragment of immunogen, termed GRG (genetically related factor), are a site of Ir gene action was investigated by using the synthetic polypeptide antigen (T,G)-A--L. It was found that T cells from (responder x nonresponder) F1 mice were stimulated by responder "macrophages" or GRF derived from these cells but not by the nonresponder macrophages of GRF from these cells. This suggests that the defect in helper cell induction in nonresponders is at the level of the presenting cell and that the macrophage factor GRF is a soluble Ir gene product. This conclusion was supported by the observation that there was normal presenting cell and GRF function in nonresponders, mouse strains such as CBA that yield helper cells and helper factor with (T,G)-A--L and have defects elsehwere.

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Suppressor cell induction in vitro. VI. Production of suppressor factors to synthetic polypeptides GAT and (T,G)-A--L from cells of responder and nonresponder mice.

The capacity of responder and nonresponder strains of mice to generate suppressor cells and factors to two antigens under MHC linked Ir gene control was investigated. Eight different H-2 types (H-2b,d,f,k,p,q,r,s) as well as seven independently derived strains (B10, BALB/c, CBA/Ca, A/St, DBA/2, P/J, SJL) were tested, and all yielded suppressor factor (SF) to (T,G)-A--L and GAT. This indicated that the genetic control of SF production was different from that of helper cell induction. Unlike previous reports of GAT suppressor extracts that GAT-specific supressor factors acted equally on both responder and nonresponder strains. As reported earlier with in vitro induced protein- (KLH) specific suppressor factors, GAT and (T,G)-A--L specific suppressor factors failed to show any genetic restriction in their function. The implications of these results for the general mechanism of Ir gene control are discussed.

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