- Volume 44, Issue 1, 1979

Rotavirus infected monkey kidney cells (LLC-MK2) have been labelled with 35S-methionine in the presence of actinomycin D. The cells have been lysed with SDS and the polypeptides separated by discontinuous polyacrylamide gel electrophoresis. Rotavirus polypeptides began to appear 4 to 5 h p.i.; incorporation was maximum at 8 h, but all the polypeptides were still being made 15 to 18 h p.i. Tissue culture adapted calf rotavirus particles were labelled with 35S-methionine and the polypeptides compared with cell associated rotavirus polypeptides. There were four inner coat, four outer coat and three non-structural polypeptides. Several of the outer coat polypeptides have altered mol. wt. on maturation. The polypeptides of rotavirus from seven species (human, pig, calf, lamb, mouse, foal and rabbit) have been compared and their mol. wt. calculated. The polypeptides fell into the same relative groupings for each virus, but there were variations in the mol. wt. of most comparable polypeptides. The polypeptides of tissue culture adapted and non-adapted calf rotavirus from the same original isolate varied only in one of the non-structural polypeptides.

A comparative study of the expression of retrovirus-associated 8S RNA was made in different mammalian cells. This RNA is found in cultured cells from all mammalian species analysed but its expression varies. An increase of 8S RNA is observed in sarcoma virus-transformed cells as compared to control uninfected cells or cells infected with leukaemia virus. No increased quantities of 8S RNA were detected in cells transformed by SV40 or by methylcholanthrene. These data show that the level of 8S RNA was augmented following transformation by sarcoma viruses.

The role of antiviral antibody in controlling the acute and latent phases of herpes simplex virus (HSV) infection in sensory ganglia of mice was studied in vitro and in vivo. Organ cultures of ganglia inoculated in vitro with HSV produced infectious virus for at least 3 weeks. In the presence of antiviral antibody, the titre of virus was markedly reduced, but the infection was not eliminated. Similarly, passive administration of antibody to HSV-infected immunodeficient (nude) mice reduced the virus titre but did not eliminate the acute phase of the ganglionic infection. Suppression of the cell-mediated immune response in latently infected immunocompetent mice by treatment with cyclophosphamide and/or X-irradiation resulted in reactivation of HSV in up to 70% of the animals. Reactivation was demonstrated by recovering infectious virus in cell-free homogenates of ganglia and eye globes and by finding virus antigens in ganglia by immunofluorescent staining. Reactivation occurred both in vitro and in vivo in the presence of high concentrations of neutralizing antibody. It is concluded that antibody alone is not sufficient to eliminate the acute phase of the ganglionic infection and that cytotoxic agents known to suppress the host’s cellular immune response can reactivate virus in the presence of neutralizing antibody.

Herpes simplex virus type 1 (HSV-1) interfered with the adsorption of subsequently added homotypic but not heterotypic HSV, suggesting that the cellular receptors involved were type-selective. Both infective and u.v.-irradiated virus could block the attachment of virions to cellular surface receptors. The adsorption rate was studied by assaying non-adsorbed infective virus remaining in the fluid medium and cell-associated 3H-thymidine labelled HSV, and HSV mutants assayed in presence of phosphonoformic acid (PFA). The adsorption profiles indicated that GMK AH-1, Vero and SIRC cells all exhibited more HSV type 1-than type 2-selective receptors while HeLa S3 cells displayed more receptors with affinity for type 2 than for type 1. On HEp-2 and human embryonic lung cells HSV type 1- and type 2-selective receptors were about equally represented.

The human lymphoblastoid cell line, Namalwa, can be cultured in serum-free media to cell densities of 3 to 4 × 106 cells per ml. These cultures produce up to 10000 units of interferon per ml when induced with Newcastle disease virus, strain B1. Maximum accumulation of interferon was obtained at approx. 13 h post-induction.

After treatment with Sendai virus, Namalwa cells form large amounts of interferon. RNA extracted from treated whole cells or from their polysomes was injected into Xenopus laevis oocytes and the interferon formed was titrated. The results show that the amount of interferon mRNA was maximal by 9 h after treatment of the cells with Sendai virus and then declined. Sucrose gradient centrifugation of the mRNA gave substantial purification and showed that its size was 12 S.

The antigens from cytolytic infections of HEp2 cells by type I simian foamy virus produced two multicomponent precipitation lines when tested by immunodiffusion with the homologous hyperimmune rabbit antiserum. The antigens obtained from a non-productive infection of MK5 virus in HEp2 cells produced only those precipitation lines which corresponded with the inner lines obtained from the cytolytic infection. Similarly, hyperimmune rabbit antiserum against antigens extracted from the persistent infection lacked the antibody which was responsible for the outer lines of precipitation. Indirect immunofluorescence with acetone-fixed and unfixed cells using the homologous and heterologous sera confirmed the absence of antigens in the persistent infection and showed that an antigen is produced in the persistently infected cells which is either absent or present in very small amounts in cytolytically infected cells. Neutralization experiments and ether treatment suggested that the missing antigens in the persistent infection were the envelope components of foamy virus. It is proposed that the persistent infection has properties in common with some infections by RNA tumour viruses.

Crude human lymphoblastoid interferon has less ribonuclease activity than equivalent primary leukocyte interferon and ribonuclease was eliminated when it was purified. The methods used differed from those that had failed to eliminate similar activity from leukocyte interferon. This result makes it unlikely that exogenous ribonuclease plays a major role in the antiviral action of interferon preparations.

A previously described revertant cell line (K-BALB SR1212), derived as a single cell clone from a clonal line of murine fibroblasts (K-BALB) transformed by a nonproductive infection with the Kirsten strain of murine sarcoma virus, has normal morphology and growth kinetics and, unlike the transformed parent cell line, lacks a sarcoma virus that can be rescued. We report here that this reversion correlates with low to undetectable levels of expression of cellular Ki-MSV-specific RNA and a reduction of proviral sequences in the cell DNA to a level equivalent to that found in the uninfected BALB cells with a normal phenotype. The data indicate that phenotypic reversion has occurred as a consequence of the loss of part or all of the sarcoma provirus, either by chromosomal rearrangement or provirus excision.

H3N2 strains of influenza A isolated from swine in Hong Kong were compared with human strains of H3N2 influenza A variants in reciprocal HI tests using ferret sera. One isolate from swine was indistinguishable from A/Hong Kong/68, one set of viruses isolated in 1976 and 1977 was most related to A/Hong Kong/68 but was not identical to it, two isolates from 1976 were ‘bridging strains’ that cross-reacted equally with the contemporary variants A/Victoria/3/75 and A/Texas/1/77, similarly to a small number of recent human isolates, and two isolates from 1977 were similar to A/Victoria/3/75. These general relationships were supported by neuraminidase inhibition tests. The findings confirm and extend previous results indicating that swine may be a reservoir of old and novel variants of influenza A H3N2 strains related to those that infect man.

Single-stranded polynucleotide preparations [tRNA, poly(rI) plus poly(ho5C)-copolymer] which protect mice against picornavirus infections without inducing interferon, protected mice equally against infection with an interferon-sensitive mutant (IS-1) of Mengo virus and with wild-type virus (IS +). Poly(rI).poly(rC) and mouse macrophage interferon [i.e. serum from mice treated with poly(rI).poly(rC)] protected mice equally against infections with the two viruses, but fibroblast interferon protected better against infection with the interferon-sensitive mutant than with the wild-type virus. These and other results indicate that: Mengo virus has a genetic locus affecting sensitivity to fibroblast but not macrophage interferon; these two types of interferon have different mechanisms of action against Mengo virus infections in mice; Mengo virus genes controlling sensitivity to fibroblast interferon may modulate disease since infection in vivo induces only fibroblast interferon; the antiviral activity of the single-stranded polynucleotides is unlikely to be mediated by induction of either macrophage or fibroblast interferon.

In cultures of Ly cells treated with 10 or 30 reference units/ml of mouse interferon there was a 30 to 200 times reduction in the production of infectious vesicular stomatitis virus (VSV); virus particle production, as measured by VSV particle associated virus RNA, virus protein, and virus transcriptase, was inhibited by, at most, six times. These results suggested that interferon-treated cells produce VSV particles with low infectivity and resemble the findings in interferon-treated cells infected with murine leukaemia viruses.

Various strains of vaccinia, variola, whitepox, monkeypox and cowpox viruses were examined for their capacity to induce a specific early antigen detectable on the surface of infected cells. The Elstree strain of vaccinia, two strains of variola minor and white variants of cowpox and monkeypox viruses lacked the capacity to induce the antigen. Variation of the parent cowpox and monkeypox viruses to white variants was always accompanied by the loss of the antigen-inducing capacity.