The Identification of Murine Peyer’s Patch T Cell-Derived Factors which Enhance IgA Responses
The humoral immune response to ingested antigens is dominated by antibody of the IgA isotype. In the mouse as in other higher mammals the major inductive site of secretory IgA responses is the Peyer’s patch (1). Work in this and other laboratories has indicated that regulatory T cells (2) and accessory cells (3) within the PP microenvironment play an important role in controlling the isotype specificity of mucosal immune responses. We have previously shown that treatment of whole PP cell populations, derived by enzymatic disaggregation, with the oxidative mitogen sodium periodate results in the formation of cell clusters enriched in L3T4+ T cells and dendritic cells (DC) (4). Co-culture of these cluster cells with either PP B cells or bone marrow mononuclear cells enhances polyclonal IgA production(5). Furthermore, the IgA inductive effect of the cluster cells is also found in 7 day culture supernatants of isolated clusters, indicating that the effect is mediated by soluble factors. T cell hybridomas have been produced from cluster T cells and the culture supernatants screened for IgA enhancing activity on whole PP cultures. Culture supernatants which showed IgA enhancing activity were tested for well characterized lymphokines including IL-2, IL-4 and IL-5. Attempts were also made to generate DC hybridomas by fusion of DC enriched cluster populations with a HAT sensitive subclone of the macrophage line P388D1. These initial fusions however, produced further T cell hybridomas as demonstrated by expression of surface Thy 1.2 antigen and the production of multiple T cell lymphokines. Culture supernatants from several of these hybridomas also showed potent IgA enhancing activity when added to whole PP cell cultures or LPS stimulated spleen B cell cultures. In all cases, hybridoma supernatants exhibiting IgA enhancing activity were positive for IL-4 and IL-5. To further investigate the role of these lymphokines in IgA synthesis, LPS stimulated spleen B cells and nonstimulated PP B cells were cultured with recombinant IL-4 and/or IL-5 to see if the IgA enhancing activity was associated with these molecules.
KeywordsBone Marrow Mononuclear Cell Sodium Periodate Cell Differentiation Factor Enzymatic Disaggregation P388D1 Cell Line
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