Staining of surface antigen on migrating lymphocytes in the chemotaxis membrane. I: Complement C5 attracts helper/inducer T cell much more than suppressor/cytotoxic T cell and NK cell.

Abstract

We have measured various chemotactic activities for neutrophils (1-3), lymphocytes (4), macrophages (5) and eosinophils (6-11) by the modified Boyden chamber method. In the conventional method, it is necessary to purify the indicator cells in upper chamber because it is impossible to distinguish inflammatory cells on the chemotactic membrane. For example, when we estimate T lymphocyte chemotaxis, we collected nylon-wool nonadherent peripheral blood lymphocytes (PBL) and confirmed their purity by sheep erythrocytes rosette formation assay. Pohajdak et al (12) evaluated NK cell chemotaxis by using large granular lymphocytes (LGL) for indicator cells. LGL fractionation was confirmed by monoclonal antibodies, HNK-1, OKT11, OKT3, OKT4, OKT8, OKM1, and MO2. When we arranged the study to evaluate chemotaxis of lymphocyte subsets which were divided by surface antigens, the conventional method needed a large quantity of PBL for the indicator cells. Moreover, one or two lymphocyte subsets had to be obtained from an identical PBL donor. This was time consuming. So, we tried to identify migrating lymphocytes in the chemotaxis membrane by their surface antigens. In this communication, we report that the staining of surface antigens on migrating lymphocytes in the chemotaxis membrane is effective and useful for evaluation of lymphocyte subset chemotaxis. It prompted us to report the new findings about modification of lymphocyte subset chemotaxis by C5a.