In vitro study of immune tolerance induced by CTLA4–Ig in bone transplantation: The effect on cell proliferation stimulated by lymphocytes and bone supernatant

Guo Chuanyou, Dai Kerong and Tang Tingting
Dept. of Orthopaedic Surgery, the Ninth People³s Hospital, Shanghai Second Medical University, Shanghai, China

To clarify the effect of CTLA4–Ig on immune rejection of bone grafts, we observed the effect of CTLA4–Ig on lymphocyte proliferation of BALB/C mice stimulated by lymphocytes and bone supernatant of C57BL/6 mice. The splenic lymphocytes and bone supernatant of C57BL/6 mice, as the stimulator cells and stimulator antigens, were cultured in vitro with the splenic lymphocyte of BALB/C mice. At the same time, CTLA4–Ig at a dose of 5, 10 or 20 µg/ml and L6 (as control) at 20 µg/ml were added. Six days later, the incorporation of 3 H-TdR was determined. Results indicated that CTLA4–Ig at a dose of 5, 10 or 20 µg/ml significantly inhibited the cell proliferation stimulated by lymphocytes and bone supernatant of C57BL/6 mice. The effect was non-cytotoxic. L6 showed no significant inhibition of cell proliferation. CTLA4–Ig can efficiently block the proliferation of alloresponsive T cell stimulated by lymphocytes and bone supernatant of C57BL/6 mice. This study provides a basis for further study of CTLA4-induced immune tolerance of bone grafts.

Key words: CTLA4–lg, immune tolerance, lymphocyte, bone supernatants, mouse

 
Journal of Orthopaedic Surgery 2000, 8(1):79–82
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last updated 1 Dec, 2002