Effects of cryopreservation of immune responses. XI. Heightened secretion of tumor necrosis factor-alpha by frozen human peripheral blood mononuclear cells

摘要

In view of the wide usage of frozen PBMCs as stem cell support following high-dose chemo- and/or radiotherapy, and the pleiotropic activities of tumor necrosis factor-alpha (TNF-alpha), the influences of freezing and radiation on LPS-induced TNF-alpha production by human peripheral blood mononuclear cells (PBMCs) were studied. Frozen PBMCs secreted significantly larger quantities of TNF-alpha than fresh cells. Blocking of endogenous IL-10 by neutralization with anti-IL-10 monoclonal antibody resulted in further augmented and prolonged secretion of TNF-alpha by both the fresh and frozen cells. In contrast, addition of exogenous IL-10 to LPS-stimulated cultures inhibited TNF-alpha secretion. In vitro irradiation had an inconsistent effect on TNF-alpha production by the fresh PBMCs. Taken together, these results suggest that the endogenously hypersecreted TNF-alpha is indirectly responsible for the previously reported elevated IL-1-, IL-6-, and IL-10-secreting capabilities of frozen PBMCs. They also indicate that the TNF-alpha induced IL-10 and then down-regulates the monocytes from further TNF-alpha secretion. Considering the vital role played by TNF-alpha in antimicrobial and antitumor activities, in the immune system, and in the pathogenesis of many acute and chronic diseases, the abilities of frozen cells to produce large quantities of TNF-alpha in response to infectious agents could have profound impact on patients receiving such frozen PBMCs as stem cell support following myeloablative therapies.