Definite spontaneous cell-mediated cytotoxicity and Hnk-1 cells in the human large intestine

Abstract

Spontaneous cell-mediated cytotoxicity (SCMC) and the marker of natural killer (NK) cells mediating SCMC of the human large intestine were studied. Lamina proprial lymphoid cells (LPL) were isolated by sequential dithiothreitol-EDTA-collagenase treatment of the gut specimen. SCMC was measured by the chromium release method. Target cells included P4788 in monolayer, a cell line derived from colon cancer, Chang cells in monolayer, and K562 in suspension. Target cells in monolayer including colon cancer cell line were chosen because they were thought to be more appropriate to assess SCMC for lymphoid cells in the solid organ. While lower compared to cytotoxicities (CT) by peripheral blood lymphoid cells (PBL), define CT were observed in LPL against all three targets. NK cells marker was studied both on LPL by an indirect fluorescent antibody method and on the gut tissue by indirect immunoperoxidase staining using anti HNK-1 monoclonal antibody which defines virtually all NK cells. HNK-1 positive (HNK-1 +) cells were identified in both methods. HNK-1 + cells were observed in the epithelium, lamina propria, and lymph follicle with or without germinal centers. These results clearly demonstrated the presence of SCMC and HNK-1 + cells in the human large bowel.

NK activities and HNK-1+ cells in familial polyposis coli.

“…Similar numbers of HNK-1+ cells were observed among the epithelium, the lamina propria and the lymph follicles in FPC compared to those of normal large bowel mucosa (Chiba et al 1986). The germinal center in the lymph follicle in FPC also had numerous HNK-l+ cells as normal large bowel.…”

“…Numbers of positive cells (HNK-1+ cells) were expressed as cells/mm2 in the lamina propria, as cells per 1,000 epithelial cells in the epithelium, and as cells per thousand lymphoid cells in the lymph follicle. HNK-1+ cells in FPC were compared to our previous results of HNK-1+ cells in sixteen normal large bowel tissues (Chiba et al 1986). …”

“…Cytotoxicity against K562 cells, Chang liver cells, and P4788 cells were assayed as described before (Chiba et al 1986). The methods are described briefly.…”

“…P4788 cells were derived from pleural effusions due to metastasis from colon cancer ( Indirect immunofluorescence antibody method NK cells in PBL were identified with HNK-1 monoclonal antibody (kindly supplied by Dr. T. Abo, Tohoku University) by an indirect immunofluorescence antibody method as described before (Chiba et al 1986). …”

“…Tissue fixation, tissue mounting, and immunoperoxidase staining were performed as described before (Chiba et al 1986). Briefly, tissue pieces were fixed with periodate lysine-4% paraformaldehyde.…”