Requirements for stimulation of T cell responses against virus-infected cells: nature of ectromelia virus-infected cells capable of stimulating cytotoxic T cells in a secondary response in vitro.

The nature of infected stimulator cells in the in vitro secondary cytotoxic T cell response to ectromelia infection was investigated. It was found that macrophages were better stimulator cells than spleen cells. B cells (Ig-positive cells) were superior to T cells (Ig-negative cells) both on a relative proportion and on a cell-to-cell basis. Concanavalin A and lipopolysaccharide-stimulated lymphocytes were also effective stimulator cells but appeared to be slightly inferior to spleen cells. Spleen cells depleted of Ia-positive cells were markedly inferior to normal spleen cells as stimulators. It was also found that primary and secondary cytotoxic T cells were largely Ia-negative. These findings are discussed in relation to the likely events during T cell responses to infection in vivo.     

Induction of cell surface expression of HLA antigens by human IFN-gamma encoded by recombinant vaccinia virus

A recombinant vaccinia virus (VV) encoding human IFN-gamma (VV-huIFN-gamma) was constructed and its effects on MHC Ag expression in human and murine cells in vitro analyzed by flow cytometry. At high multiplicities of infection (5 pfu/cell) the IFN-gamma expressed by vaccinia was not able to overcome the profound decrease of MHC concentration, normally associated with VV infection, in any of the cells tested. However, at successively decreasing multiplicities of infection, a gradual increase in MHC class I concentration above control levels was observed in human 143B cells but not in murine L929 cells, thus indicating that the species specificity of IFN-gamma is preserved in VV-huIFN-gamma-infected cells. We infer from these data that the IFN-gamma secreted by infected 143B cells is able to exert an MHC upregulating effect on uninfected cells in the vicinity. Antiviral activity of the IFN-gamma expressed by the virus was also assessed. Pretreatment for 24 to 48 h of human 143B cells with IFN-gamma containing supernatants had a significant antiviral effect comparable to rhuIFN-gamma. However, when added 1 h after virus infection, antiviral activity was much less evident. Also, the IFN-gamma secreted by infected 143B cells in monolayers infected at low multiplicity did not efficiently inhibit spread of infection to other cells in the vicinity.     

The appearance of surface H-2 antigens on fetal and postnatal murine hepatocytes in vivo and in vitro: A comparative study of MHC control

We have collected data on the kinetics of in vivo development of H-2^b antigens in fetal and postnatal hepatocytes from days 15–89 postcoitum (pc) in C57BL/10 mice using a sensitive immunoferritin labeling method combined with electron microscopy. We compared these data with data on the kinetic development of H-2^b antigens on fetal hepatocytes cultured from days 15, 16, and 17 pc onwards. Using the same techniques, we also compared the surface concentrations of H-2 antigens on the hepatocytes of pregnant and age-matched male and virgin female mice at day 110 pc. We found that the surface concentration of H-2 antigens on fetal and postnatal hepatocytes increased with time but that the birth event was associated with an increase in H-2 antigen expression from 60–80 ferritin grains per meter (F/m) to 190 F/m followed by a decrease to 115 F/m within 72 h of birth. The concentration of antigens on postnatal hepatocytes increased gradually and reached a maximum of 640 F/m on day 41 pc. By day 89 pc, however, this level had decreased 433 F/m, which was not significantly different from that found in day 110 pc males and virgin females. In contrast to previous findings, we found that hepatocytes cultured from days 15, 16, and 17 pc exhibited a large increase in H-2 surface antigen concentration within 48 h from 60–80 F/m to 1500–2200 F/m. After day 2, the H-2 concentration decreased and by days 14–17 in culture it was not significantly different from day 110 pc male and virgin female levels (485 F/m). Lastly, we found that the H-2 concentration on hepatocytes from pregnant mice was increased significantly (725 F/m) compared with the concentration of hepatocytes from day 110 pc males or virgin females. We postulate that the control of cell-surface major histocompatibility complex antigen concentration is achieved by a combination of intra and extracellular mechanisms and we discuss how this might be of benefit to the animal in vivo.