Morphological study of cytotoxicity produced by PSK-induced polymorphonuclear leukocytes (PMNs) and Nocardia rubra cell wall skeleton.

Abstract

The morphologic changes in PMNs induced by an i.p. injection of PSK, a polysaccharide from the mycelia of Coriolus versicolor, and tumor cells undergoing cell death, were evaluated by immunohistochemical staining and electron microscopy. Male C3H/He mice, 8-10 -weeks old, received an i.p. injection of 125 mg/kg of PSK. Their PMNs were obtained 6 h after the PSK injection by peritoneal lavage. N-CWS (Nocardia rubra cell wall skeleton) was added at the start of the chromium release assay using the MM46 mammary carcinoma cell line, which is syngeneic to C3H/He mice, as target cells. During the cytotoxic assay, the cells were fixed at various time points. The MM46 cells expressed ICAM-1 while the PMNs expressed both ICAM-1 and LFA-1 as determined by immunohistochemical staining and immunoelectron microscopy using anti-ICAM-1 and anti-LFA-1 antibodies. PMNs with ruffle-like microvilli adhered to the MM46 tumor cells 30 min after the addition of N-CWS. Immunoelectron microscopic findings suggested that the adhesion molecules were LFA-1 on the PMNs and ICAM-1 on the MM46 tumor cells, but cell fusion between the PMNs and tumor cells was not observed. The MM46 tumor cells gradually lost their microvilli, which showed cell damage, and died 6-7 h after the addition of the N-CWS. This time course of tumor cell death is compatible with the results of the cytotoxic assay. Pretreatment of PMNs by anti-LFA-1 antibody suppressed 1% lysis of MM46 tumor cells from 90% to 10% (p < 0.01). These data suggest that adhesion molecule on the surface of PMNs such as LFA-1 might play an important role on signal transduction of these PMNs cytotoxic function in this experimental system.

• Authors: H Kata, M Inoue, S Mukai, Y Kawahito, T Yoshida, K Asai, S Kimura, A Hashiramoto, Y Yamamura, H Sano, S Sugino, M Kondo
• Journal: Biotherapy (Dordrecht, Netherlands) (Biotherapy) Vol. 9 Issue 4 Pg. 229-39 ( 1996) ISSN: 0921-299X [Print] Netherlands
• PMID: 9012542 (Publication Type: Journal Article)

Chemical References

• Adjuvants, Immunologic
• Cell Wall Skeleton
• Lymphocyte Function-Associated Antigen-1
• Proteoglycans
• Intercellular Adhesion Molecule-1
• polysaccharide-K

Topics

• Adjuvants, Immunologic (pharmacology)
• Animals
• Cell Adhesion
• Cell Wall Skeleton (pharmacology)
• Cytotoxicity, Immunologic
• Immunohistochemistry
• Intercellular Adhesion Molecule-1 (analysis)
• Lymphocyte Function-Associated Antigen-1 (analysis)
• Male
• Mice
• Mice, Inbred C3H
• Microscopy, Electron
• Microscopy, Immunoelectron
• Neutrophils (drug effects, immunology, ultrastructure)
• Nocardia (ultrastructure)
• Proteoglycans (pharmacology)
• Tumor Cells, Cultured