An IL-3-induced splenic NC-1.1+ mast cell line mediates natural cytotoxicity independent of TNF-alpha.
Long-term culture of mouse spleen cells in IL-3-conditioned medium induced a stable mast cell line. This mast cell line (MCL) which could not be cloned expressed NC-1.1, a receptor on cells which mediate natural cytotoxicity (NC), and CD32/CD16 but no markers of T cells, B cells, macrophages, or NK cells. The MCL cells were large and granular, with abundant cytoplasm and >90% stained with Alcian blue, a mast cell-specific stain. Probing total RNA with cDNA encoding a mast cell-specific proteinase mMCP-5 identified the approximately 1-kb mMCP-5 transcript which was confirmed at the protein level. MCL cells mediated very high NC against WEHI-164 which increased with time in culture and which was blocked by anti-NC-1.1 but not by anti-TNF-alpha. When incubated with WEHI-164 tumor cells, MCL cells showed transient induction of TNF-alpha mRNA, but no detectable surface protein. Thus long-term culture of murine spleen cells in IL-3 induces mast cells which express the NC-1.1 receptor of cells which mediate NC, and utilize a cytotoxic effector mechanism which is not TNF-alpha.
Shirzadeh
H
H
Discipline of Surgical Science, Faculty of Medicine and Health Sciences, University of Newcastle, New South Wales, Australia.
Clarke
G R
GR
McNeil
H P
HP
Wang
H
H
Burton
R C
RC
Smart
Y C
YC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015415:Biomarkers; D002460:Cell Line; D017077:Culture Media, Conditioned; D003602:Cytotoxicity, Immunologic; D016130:Immunophenotyping; D007376:Interleukin-2 / Q000276:immunology*; D008407:Mast Cells / Q000276:immunology*; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D008808:Mice, Inbred CBA; D013154:Spleen / Q000166:cytology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D017077:Culture Media, Conditioned; D007376:Interleukin-2; D008565:Membrane Proteins; D014409:Tumor Necrosis Factor-alpha