Host Cell Glycoprotein Research Articles

Human natural killer cell lysis of virus-infected cells. Relationship to expression of the transferrin receptor.

Natural killer (NK) cells lyse tumor and virus-infected cells yet the nature of the target structure they recognize is unknown. A normal host cell glycoprotein, the transferrin receptor (TfR), has been proposed as a target structure on tumor cells. We therefore investigated whether changes in the number or physiological recycling of the TfR, consequent on virus infection, were related to the differential susceptibility of virus-infected cells to NK lysis. There was a direct correlation between TfR expression, susceptibility to NK lysis and ability to act as cold target competitors, for human fibroblasts infected with RNA and DNA viruses (cytomegalovirus, herpes simplex, polio, vaccinia and Semliki Forest virus). The NK lysis of human cytomegalovirus-infected fibroblasts was studied in more detail. NK lysis was increased coincident with human cytomegalovirus early antigen expression and this susceptibility to lysis was associated with increased total and recycling TfR but only a slight increase in surface TfR expression. In addition, susceptibility of uninfected human fibroblasts to NK lysis directly correlated with TfR number. However, we were unable to inhibit NK lysis by either excess iron-saturated Tf or affinity-purified TfR. We conclude that there is a direct correlation between total TfR expression and susceptibility to NK lysis of human virus-infected cells; however, the NK target structure on virus-infected cells is probably not the TfR itself.

Sodium butyrate selectively inhibits host cell glycoprotein synthesis in human fibroblasts infected with cytomegalovirus

Host cell as well as viral DNA synthesis in human fibroblasts infected with human cytomegalovirus was found to be largely resistant even to high concentrations of sodium butyrate. Likewise, production of viral progeny was reduced by 1-2 orders of magnitude but not abolished. On the other hand, the drug allowed (modified) glycosylation only of viral polypeptides whereas that of host proteins was suppressed. Immunofluorescence studies on living cells suggested that butyrate may interfere with processing and intracellular transport of virus-specific surface membrane antigens.

Specific selection of host cell glycoproteins during assembly of murine leukaemia virus and vesicular stomatitis virus: presence of Thy-1 glycoprotein and absence of H-2, Pgp-1 and T-200 glycoproteins on the envelopes of these virus particles.

Using the indirect immunoelectron microscopy technique, it was investigated whether during assembly of murine leukaemia virus (MuLV) and vesicular stomatitis virus (VSV), the glycoproteins (gp) Thy-1, H-2, Pgp-1 and T-200 present on the surface of BW5147 and BuEL4 leukaemia cell lines were incorporated into the virus envelopes. This work was done mainly with monoclonal antibodies against these gps to exclude the presence of antibodies against endogenous MuLV present in conventional mouse antisera. Thy-1 gps were incorporated into MuLV and VSV envelopes. In contrast, H-2, Pgp-1 and T-200 gps were excluded from the budding of both virus particles. To study whether the presence of Thy-1 gps on the viral envelopes is due to some lateral affinity of this molecule with viral gps, the physical association of Thy-1.1 antigens and MuLV antigens was studied with antibody-induced redistribution of both antigens on the BW5147 cell surface. Antibody-induced patching of the viral antigens did not result in co-patching of the Thy-1.1 antigens. In the reciprocal tests no co-redistribution of viral antigens with Thy-1.1 antigens was seen. These studies show that the presence of Thy-1.1 gp on the MuLV envelope cannot be due to a lateral affinity of this molecule with viral gps and that a selection of surface gps takes place during assembly of MuLV and VSV.

The synthesis of glycoproteins in human melanoma cells infected with varicella-zoster virus

The synthesis of glycoproteins has been investigated in cultured human melanoma cells before and after infection with varicella-zoster virus (VZV). The glycosylated polypeptides were labeled with tritiated precursors of the sugar moiety and were analyzed by polyacrylamide gel electrophoresis. When the electrophoretic profiles of uninfected and infected cell lysates were compared, a marked reduction in host cell glycoprotein synthesis was evident during the late phase of viral infection. In addition, at least three infected-cell specific (ICS) glycosylated polypeptides were detected. After immune precipitation of [ 3H]glucosamine-labeled VZV cell extracts with human and rabbit VZV antisera, a total of five ICS glycoproteins were identified and designated gp 45, gp 62, gp 88, gp 98, and gp 118, according to their apparent molecular weights (× 10 3). Four of the five polypeptides (but not gp 88) were visualized after immune precipitation of infected cell extracts labeled with [ 3H]fucose. The ICS glycopeptides corresponded in molecular weight to [ 35S]methionine-labeled polypeptides detected in a virus-enriched fraction obtained by centrifugation of sonically disrupted infected cells in a combination density-viscosity gradient. A prominent ICS high-molecular weight (∼150,000) nonglycosylated polypeptide was identified also in the latter fraction.