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Expression of perforin and membrane-bound lymphotoxin (tumor necrosis
factor-beta) in virus-specific CD4+ human cytotoxic T-cell clones
M Yasukawa, Y Yakushijin, H Hasegawa, M Miyake, Y Hitsumoto, S Kimura, N Takeuchi and S Fujita
First Department of Internal Medicine, Ehime University School of Medicine,
Japan.
In an attempt to clarify the mechanisms of cytotoxicity mediated by CD4+
cytotoxic T lymphocytes (CTL), the expression of perforin and
membrane-bound lymphotoxin (LT) (tumor necrosis factor-beta) in herpes
simplex virus (HSV)-specific CD4+ human cytotoxic and noncytotoxic T- cell
clones was examined. Three HSV-specific CD4+ human CTL clones that showed
HLA-DR-restricted cytotoxicity and proliferative response were established.
The cytotoxicity of these clones in 5-hour 51Cr release assays was found to
be mediated by the directional target cell lysis and not by the release of
cytotoxic soluble factors, ie, "innocent bystander" killing. Northern blot
analysis showed that messenger RNAs for perforin and LT, which were both
considered to be important mediators for cytotoxicity of CD8+ CTL and
natural killer cells, were abundantly expressed in HSV-specific CD4+ CTL
clones. Expression of perforin in the cytoplasm of CD4+ CTL clones was also
detected by immunohistochemical staining using a monoclonal antibody
against perforin. In addition, LT bound to the cell surface of CD4+ CTL
clones was detected by flow cytometry. In contrast, little or no expression
of perforin and LT was detected in three HSV-specific CD4+ noncytotoxic T-
cell clones. Although the cytotoxicity mediated by lymphokine-activated
killer cells was partly inhibited by addition of anti-LT antibody, it did
not show any effect on the cytotoxicity of HSV-specific CD4+ CTL clones. In
addition, it was found that cytotoxicity mediated by these CD4+ CTL clones
was Ca2(+)-dependent. These data thus suggest that perforin and
membrane-bound LT are both expressed in HSV-specific CD4+ CTL, although
perforin might be the more important mediator in short- term culture.
Volume 81,
Issue 6,
pp. 1527-1534,
03/15/1993
Copyright © 1993 by The American Society of Hematology
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