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Pulmonary Research Group, Department of Medicine, University of Alberta, Edmonton, Alberta, Canada
CD8, a marker largely restricted to subsets of T lymphocytes and NK
cells, was detected on freshly isolated rat peritoneal mast cells
(PMC). Using flow cytometry, Percoll-enriched rat PMC (
98% purity)
were positive for the hinge region of CD8
(67.5 ± 9.5%; Ab
OX8) and CD8ß (27.8 ± 2.3%; Ab 341). CD8+ PMC
consisted of two populations, CD8+ (22.5%) and
CD8+ß+ (15.9%). Interestingly, G28, an Ab
that identifies the IgV-like region of CD8 on T lymphocytes, did not
bind PMC, suggesting that PMC CD8 is distinct from that on T
lymphocytes. Moreover, a similar pattern of Ab positivity for CD8 was
observed on a rat mast cell line, RBL 2H3. The presence of CD8
immunoreactivity on rat PMC was further confirmed by confocal
microscopy. In situ reverse-transcription PCR and reverse-transcription
PCR analysis demonstrated that PMC contained mRNA transcripts encoding
CD8. In functional studies of CD8 on PMC, both TNF- and nitric
oxide production were induced by OX8 (CD8) and 341 Ab (CD8ß) in a
dose-dependent manner. However, neither OX8 nor 341 induced histamine
secretion from PMC. Ag-induced secretion of TNF-, nitric oxide, and
histamine was not affected by OX8 or 341 Abs, suggesting that there are
distinct signaling mechanisms mediated by CD8 and Fc
RI. These
results indicate that rat PMC express functional CD8 molecules that may
be distinct from those of T lymphocytes. The difference suggests there
is a ligand other than MHC class I for mast cell
CD8.
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