Summary. In the present study, we demonstrated that a close relationship exists between hepatitis C virus (HCV) infection of peripheral blood mononuclear cells (PBMCs) and cell-surface Fas expression in patients with hepatitis C, and showed the possibility of PBMCs apoptosis via a Fasmediated system. The expression of Fas on PBMCs was found by flowcytometric analysis to be significantly increased in these patients. In addition, the treatment of patients' PBMCs with anti-Fas antibody induced cell death, with nuclear condensation and fragmentation and cellular DNA fragmentation. These data indicate that the patients' PBMCs expressed a large amount of functional Fas on the cell surface and were susceptible to stimulation against Fas, causing apoptotic cell death. We then quantified the serumsoluble Fas ligand (sFasL), which was known to bind to Fas and induce the apoptotic signals into the sensitized cells.The patients' serum sFasL levels were significantly higher than those of normal subjects and showed a good negative correlation with their PBMC number. To demonstrate the correlation between Fas expression and HCV infection, nested reverse transcriptase polymerase chain reaction (RT-PCR) was performed to detect HCV RNA. Interestingly, HCV RNA was preferentially detected from Fas-positive cells but not from Fas-negative cells, which had been isolated from PBMCs by magnetic beads. These results suggest that HCV infection of PBMCs might induce Fas expression and additional stimulation such as sFasL might induce apoptosis in these Fas-expressing cells. These mechanisms, in addition to hypersplenism, may explain the decrease in the number of PBMCs observed in patients with chronic hepatitis C.
Electrophoresis revealed two cases of malignant lymphoma that each contained three M-proteins (IgM lambda.lgG kappa.lgG lambda and IgM lambda.IgM kappa.lgG kappa) in the sera. To determine cellular origin of each M-protein, atypical lymphoid and plasmacytoid cells of both cases were examined by electron microscopy. Atypical lymphoid and plasmacytoid cells possessed rough endoplasmic reticula (RERs) in varying degrees, as seen by conventional electron microscopy, and showed double-stainability for plural antibodies against immunoglobulins following double stainings of immunoelectron microscopy using immunogold staining. Rabbit antibodies against human IgM, lgG, free kappa-light chain and free lambda-light chain were used for the immunoelectron microscopic staining. By the double staining method, plural immunoglobulins, IgM/IgG, IgM/free kappa, IgM/free lambda, IgG/free kappa, IgG/free lambda and free kappa/free lambda, were simultaneously detected in varying degrees in the Golgi area, RERs, and dense bodies of lymphoid and plasmacytoid cells. In conclusion, this study directly exhibited, through electron microscopy, that plural immunoglobulins were synthesized at the same time in a single cell, and that the process of immunoglobulin synthesis in the lymphoid and plasmacytoid cells was different from that in a normal B-cell.
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