Leukocyte integrin-dependent and antibody-independent cytotoxicity of macrophage against allografts

Abstract

Macrophages (Mφs), but not T cells, infiltrating into the rejection site of either i.p. allografted Meth A (H-2(d)) fibrosarcoma cells in C57BL/6 (B6) (H-2b) mice or BALB/c (H-2(d)) skin onto B6 mice are cytotoxic against allografts with H-2(d) specificity. To determine the mechanisms of specific killing of allografts by allograft-induced Mφ (AIM), we raised ≃5,000 rat monoclonal antibodies (mAbs) against AIM and selected three of them (R1-73, R2-40 and R1-34), each of which inhibited cytotoxic activity against allografts in a dose-dependent manner. The antigens recognized by R1-73, R2- 40 and R1-34 mAbs were defined by immunoprecipitation and Western blot analyses as CD11(a), CD18 and CD11(b), respectively; and the allografts expressed CD54, a ligand of CD11(a), or CD11(b), suggesting leukocyte integrin-dependent killing. Although Ab-dependent cellular cytotoxicity has been recognized as a mechanism of specific killing by Mφs, the infiltration of AIM into the rejection site of allografts far (≃6 days) preceded the appearance of serum IgG Ab specific for the allograft. AIM exhibiting full cytotoxic activity against allografts was also induced in the transplantation site of Fcγ receptor knockout [(B6x129) F1] mice as well as B10.D2 (H-2 compatible with allograft) and B6-xid (X-linked immunodeficiency with B cell- specific defect) strains of mice. In the latter two strains of mice, the levels of serum IgG Ab to the allograft were negligible. Moreover, the cytotoxic activity of AIM against allografts was not affected by pretreatment of the cells with anti-mouse IgG serum, suggesting Ab-independent cytotoxicity.