ABSTRACT
In this study we investigated the co-stimulatory signaling capacity of diverse proteins of the extracellular matrix (ECM) for murine resting CD4+ T cells and Th1 clone cells, activated by immobilized anti-CD3 mAb. ECM proteins used in various concentrations had no effect on IL-2 production or proliferation of highly purified CD4+ T cell populations. When the preparation of CD4+ T cells contained contaminating accessory cells, IL-2 secretion and proliferation was enhanced in the presence of co-immobilized collagens or fibronectin. However, the level of proliferation attainable by added irradiated splenocytes was not reached. Using Th1 cell clone M4, enhanced production of IL-2 in the presence of immobilized ECM proteins was observed. At a submitogenic anti-CD3 mAb dose, proliferation of M4 T cells was augmented by the ECM proteins in a concentration range that optimally induced IL-2 production. IL-2R p55 was up-regulated on M4 T cells by collagen type IV and fibronectin to the same level that was induced by exogenously added IL-2, whereas added accessory cells induced a higher level of IL-2R p55 expression. Likewise, in dot-blot analysis a comparable quantity of IL-2R p55- and p75-specific transcripts was induced by collagen type IV or fibronectin and by IL-2, which was lower than that induced by antigen-presenting cells. Our data suggest that the enhanced proliferation of M4 T cells induced by ECM proteins is not the consequence of direct up-regulation of IL-2R, but appears to be due indirectly to elevated secretion of IL-2. At an optimal anti-CD3 mAb dose the collagens inhibited M4 T cell proliferation. Diminished cell surface expression of IL-2R p55 following stimulation with anti-CD3 mAb plus collagen type IV compared with anti-CD3 mAb alone was observed and may be responsible for growth inhibition.
Subject(s)
CD4-Positive T-Lymphocytes/drug effects , CD4-Positive T-Lymphocytes/metabolism , Extracellular Matrix Proteins/pharmacology , Lymphocyte Activation/drug effects , Lymphokines/metabolism , Th1 Cells/drug effects , Th1 Cells/metabolism , Animals , Clone Cells , Integrin beta1/biosynthesis , Lymphokines/drug effects , Mice , Mice, Inbred BALB C , Mice, Inbred C3H , Mice, Inbred C57BL , Receptors, Interleukin-2/biosynthesis , Receptors, Interleukin-2/geneticsABSTRACT
The murine T clone cells BK-BI-2.6.O4.1 (BI/O4.1) synthesize and express MHC class II molecules constitutively. BI/O4.1 cells are able to present various protein antigens to antigen-specific CD4 + T cells. However, a 10-fold higher concentration of antigen is needed to activate specific T cells to lymphokine secretion by BI/O4.1 cells in comparison with spleen cells or with the more homogeneous population of bone marrow-derived macrophages (BMMph). The authors tested whether the reduced antigen presentation potential of BI/O4.1 cells was augmented by transferrin-mediated uptake of the model antigen ovalbumin (OVA) coupled to human ferric transferrin. It was shown that 240-fold less OVA was sufficient to induce proliferation of an OVA-specific T-cell clone when the conjugate and not native OVA was used. The presence of ferric TF in the cultures competitively inhibited this effect of the conjugate. A similar shift in the dose-response curve to lower doses of antigen was induced by the conjugate when B lymphoma cells were used as antigen-presenting cells. BMMph and P388D1 cells processed and presented the conjugate with similar efficiency as native OVA, although both cell types exposed transferrin receptors. These data suggest that the reduced antigen presentation potential of BI/O4.1 T clone cells is due to the inefficient uptake of OVA by pinocytosis and delivery into the processing compartment.
