Michael L. Diegel

Co-ligation of the Antigen and Fc Receptors Gives Rise to the Selective Modulation of Intracellular Signaling in B Cells REGULATION OF THE ASSOCIATION OF PHOSPHATIDYLINOSITOL 3-KINASE AND INOSITOL 5′-PHOSPHATASE WITH THE ANTIGEN RECEPTOR COMPLEX

Cross-linking of the Fc receptor (FcR) to surface immunoglobulin (sIg) on B cells inhibits the influx of extracellular calcium and abrogates the proliferative signal. The mechanism by which this occurs is not well understood. In this report we show that co-cross-linking the FcR to the antigen receptor gives rise to very selective modulation of signal transduction in B cells. Co-cross-linking sIg and the FcR enhanced the phosphorylation of the FcR, the adapter protein, Shc, and the inositol 5′-phosphatase Ship. Furthermore, phosphorylation of the FcR induced its association with Ship. Cross-linking of the FcR and sIg decreased the tyrosine phosphorylation of CD19, which led to a reduction in the association of phosphatidylinositol 3-kinase. In addition, the phosphorylation of several other proteins of 73, 39, and 34 kDa was reduced. Activation of the cells with either F(ab′)2 or intact anti-IgG induced very similar changes in levels of tyrosine phosphorylation of most other proteins, and no differences in the activation of several protein kinases were observed. These results indicate that the inhibitory signal that is transmitted through the FcR is not mediated by a global shutdown of tyrosine phosphorylation but is, rather, a selective mechanism involving localized changes in the interactions of adapter proteins and the enzymes Ship and phosphatidylinositol 3-kinase with the antigen receptor complex.

In vivo administration of 15-deoxyspergulin inhibits antigen-presenting cell stimulation of T cells and NF-κB activation

15-Deoxyspergulin (DSG), a synthetic derivative of spergulin, was initially characterized for its antibiotic and antitumor effects. Recent studies have described the immunosuppressive properties of this molecule, but its mechanism of action is not clearly understood. In the study reported here, mice were treated in vivo with DSG prior to the measurement of IL-2 production and proliferation in an in vitro antigen presentation assay. At suboptimal antigen concentrations, elicited peritoneal macrophages or percoll isolated B cells from DSG-treated mice showed a 50-96% reduction in their ability to present chicken ovalbumin (cOva), cOva peptide, or superantigen (SAg) to MHC class II-matched antigen-specific primary T cells. No significant changes could be found in the cell surface expression of CD80, CD86, MHC I, MHC II, CD18, CD11b, CD40, CD25, and CD54 in antigen-presenting cells (APC) from control or DSG-treated animals. Activation with SAg of macrophages or splenocytes from DSG-treated mice revealed that there was a significant reduction in nuclear NF-kappaB levels compared to cells from untreated animals. Additionally, analysis of cytokines showed that production of TNF-alpha and IL-1beta was inhibited in cultures where macrophages from DSG-mice were used to present cOva to T cells. These results indicate that the effects of DSG in mice are not simply due to altered antigen processing or from any marked changes in cell surface antigen expression. Rather, the immunosuppression may arise from alterations in the release of one or more soluble factor from DSG-treated APC, which prevents effective antigen presentation and T cell activation.