Preparative nonlytic separation of Lyt2+ and Lyt2− T lymphocytes, functional analyses of the separated cells and demonstration of synergy in graft‐vs.‐host reaction of Lyt2+ and Lyt2− cells

作者：Michael G. Mage, Bonnie J. Mathieson, Susan O. Sharrow, Louise McHugh, Ulrich Hämmerling, Colette Kanellopoulos‐Langevin, Donald J. Brideau, Charles A. Thomas · 发表于：European Journal of Immunology · 年份：1981 · DOI：10.1002/eji.1830110312 · 被引用次数：84 · 研究领域：Immunotherapy and Immune Responses、T-cell and B-cell Immunology、Immune Cell Function and Interaction

A convenient, preparative scale, nonlytic separation of mouse T lymphocytes into Lyt2.2+ and Lyt2.2- populations is reported. Immunoglobulin-negative (Ig-) spleen cells, Ig- lymph node cells, and peanut lectin-unagglutinated (PNA-) thymocytes were incubated under sterile conditions at 0 degree C with monoclonal mouse antibody to the Lyt2.2 T cell differentiation antigen. The antibody-treated cells were washed and placed in polystyrene tissue culture dishes that had been precoated with antibody to mouse Ig. Nonadherent populations were depleted to Lyt2.2+ cells and were essentially devoid of cytotoxic T lymphocyte precursors (CTLp), but contained helper activity for in vivo T-dependent IgM, IgG and IgA antibody formation. Adherent cell populations were enriched for Lyt2.2+ cells and for CTLp. The graft-vs.-host activity of the separated, adherent (Lyt2.2+) and nonadherent (Lyt2.2-) cells in the Simonsen spleen assay in neonatal (C57BL/6 x BALB/c)F1 mice was less than of unfractionated cells, but the activity of remixed Lyt2.2+ plus Lyt2.2- cells was higher than the sum of the contributions of these cells tested separately, and equal to that of the unfractionated cells. PNA- thymocytes were also separated into Ly2.2+ and Lyt2.2- populations by fluorescence-activated cell sorting. Nonlytic separation allows the recovery of the Lyt1+2+ population, which is lost in cytotoxic elimination experiments. Under the conditions described for the plate separation, the purity of the separat...