This report confirms and extends the recent work of Pauwels et al. on a "reclustering" assay (a simple microtitration plate method) for the determination of human immuno-deficiency virus (HIV) infection of MT-4 cells. MT-4 cells, which are highly susceptible to and permissive for HIV, typically grow in clusters. In the absence of virus these cell aggregates, after dissociation by pipetting, reform into clusters within 2 to 3 hours. Growth of HIV results in an inhibition of reclustering, with an end-point some 4-5 days after initiation of infection. In cultures inoculated with 5 to 8 TCID(50) of HIV, only 2-4% of the cells remain viable after 4 days. Correspondingly, HIV antigens can be detected by immunofluorescence in more than 90% of the cells remaining in the culture. The sensitivity of the "reclustering" assay is only slightly less than that of the immunofluorescence test. A colorimetric assay is also described that employs a tetrazolium salt (designated as MTT) to measure the cytolytic effect of various dilutions of HIV; comparable virus titres were obtained. This reclustering assay now appears to offer the simplest method for titration of prototype HIV in virus stocks and when used in drug evaluation tests and for measurement of HIV neutralizing antibodies.

Cell Aggregation↗

PubMed1987

High levels of cytomegalovirus antibody in patients requiring vascular surgery for atherosclerosis.

157 caucasian male patients undergoing vascular surgery for atherosclerosis and a matched control group of patients with high cholesterol levels were screened for antibodies to cytomegalovirus (CMV) and herpes simplex virus type 1 (HSV1) and type 2 (HSV2), indicative of persistent infection. The prevalence of CMV antibodies was higher in the surgical group than in the control group (90% and 74%, respectively), and a significantly greater percentage (p less than 0.001) of surgical cases than controls had high titres of CMV antibodies (57% and 26%, respectively). Small but not significant differences in antibodies to HSV1 were observed, and there were no differences in HSV2 antibody titres. For each virus there was no correlation between antibody titre and blood levels of total cholesterol or triglycerides. It is suggested that periodically activated virus may have a role in the pathogenesis of atherosclerosis.

Adult↗

PubMed1987

Factors influencing the assay of Inoue-Melnick virus.

We describe factors influencing the assay of Inoue-Melnick virus (IMV) and practical comments for its assay. MRC-5 cells at a population doubling level (PDL) of more than 32 were more sensitive than younger cells for demonstrating cytopathic effect (CPE) of the virus. The PDL of MRC-5 cells also influenced the virus yield, particularly when the amount of viral inoculum was small. Cells at younger PDL produced less virus. The appearance of CPE and the virus yield were inhibited by increasing the concentration of NaHCO3 to 0.14% in the maintenance medium. The release of the virus into the medium was also inhibited as NaHCO3 was increased.

Bicarbonates↗

PubMed1987

Fusion as a mediator of cytolysis in mixtures of uninfected CD4+ lymphocytes and cells infected by human immunodeficiency virus.

We describe an unusual type of cytopathology in which uninfected CD4+ (helper/inducer) cells (cells expressing the human leukocyte antigen CD4) interact with cells persistently infected with the human immunodeficiency virus (HIV). Prior antigenic stimulation was not required, since CD4+ cells taken either from healthy persons without anti-HIV antibodies or from individuals with anti-HIV antibodies were capable of inducing cytolysis. Neither CD8+ (suppressor/cytotoxic) nor CD16+ (natural killer) cells mediated the reaction. Light microscopic and autoradiographic studies revealed that, prior to cytolysis, multinucleated giant cells were formed from fusions between HIV-infected cells and large numbers of uninfected CD4+ lymphocytes. These data may explain the paradox that exists in vivo in which a dramatic depletion of CD4+ lymphocytes occurs in the presence of a small number of HIV-infected CD4+ cells. These new insights into the pathogenesis of acquired immunodeficiency syndrome (AIDS) may lead to future therapeutic strategies.

Acquired Immunodeficiency Syndrome↗

PubMed1987

Nucleic acid sequences of cytomegalovirus in cells cultured from human arterial tissue.

Explore the source record for details and available documents.

Aorta, Abdominal↗

PubMed1987

Human immunodeficiency virus in brain biopsies of patients with AIDS and progressive encephalopathy.

Human immunodeficiency virus (HIV) has been identified in patients with AIDS in their circulating and tissue lymphocytes and in their monocyte-macrophages, an arm of the cell-mediated immune system. We report our findings of HIV in brain biopsies. Virus was detected in oligodendroglial and astroglial cells. Virus was also present in the brain capillaries, both in lumens and at the endothelial gaps. Virus was also found in extracellular spaces. Budding forms of the developing virus and bar-shaped nucleoids of mature virus, the diagnostic hallmark of the HIV retrovirus, were identified in brain tissue of five of seven patients with AIDS studied. We believe that HIV may enter the the brain via the vascular capillaries (through the endothelial gaps), bind to brain cells containing T4 receptors, enter the cells, and replicate. There follows a reactive hyperplasia and degeneration of oligodendroglial and astroglial cells. The oligodendroglial cells are the myelin-forming cells, and thus, the result is myelin degeneration.

Acquired Immunodeficiency Syndrome↗

PubMed1987

Antibody survey for Inoue-Melnick virus infections in Osaka and Houston.

Neutralizing antibodies to the Inoue-Melnick virus were detected in healthy adults in Osaka and in Houston. For adults, the prevalence rate for the cross-reacting type 3 virus was 13.8 per cent in Osaka and 29.1 per cent in Houston. Antibodies were also found in immune serum globulin prepared from American adults, but not in sera of Osaka children under 10 years of age. The Inoue-Melnick virus isolates so far obtained in Houston could be classified into three antigenic types, but in Japan all have been type 1. Type 3 antibody tests detect all homotypic infections plus about 82 per cent of type 1 infections. Thus, the actual prevalence rates for type 1 infections in Japan are somewhat higher than those found with the type 3 assay, namely, about 14 per cent for young adults and 24 per cent for those over 50 years of age. Further studies are needed to elucidate the natural history of this virus infection.

Adolescent↗

PubMed1987

Detection of antibody to group B adult diarrhea rotaviruses in humans.

Group B rotaviruses have been responsible for annual epidemics of severe diarrhea affecting both adults and children in China. We developed a specific and sensitive enzyme-linked immunosorbent blocking assay to detect antibody to group B rotaviruses that will be useful to assess the role of group B rotavirus infections as a cause of human gastroenteritis. We tested 219 human sera and 18 immunoglobulin pools collected from eight countries for antibodies to both group A and group B rotaviruses. Overall, a low proportion (10 of 237 or 4.2%) of sera contained antibody to group B rotaviruses. Antibody to group B rotavirus was detected in only 1 of 155 serum samples from healthy or hospitalized individuals in the United States, including patients with the chronic inflammatory bowel diseases Crohn's disease and ulcerative colitis. No antibody was detected in 15 serum samples from Australia and from an outbreak of gastroenteritis on a cruise ship or in nine immunoglobulin pools from Japan and the United Kingdom. Antibody to group B rotaviruses was detected in 8 convalescent-(but not acute-)phase serum samples from Chinese patients with group B gastroenteritis, in five immunoglobulin pools from China, in 1 of 6 serum samples from Chinese students in the United States, and in 1 each of 10 serum samples from Kenya, 20 from Thailand, and 15 from Canada. In contrast, most of these samples (226 of 237 or 95.4%) had antibody to group A rotaviruses. These results indicate that human infection with group B rotavirus has not been widespread in areas outside China. Seroconversion observed between the acute-and convalescent-phase serum samples from China also suggests that infections with this virus are primary infections. Continued surveillance for this new group of rotaviruses should determine whether the many susceptible people become infected of whether other factors influence the severe pathogenicity of human infections with these viruses in China.

Adolescent↗

PubMed1987

Integrated state of subgenomic fragments of hepatitis B virus DNA in hepatocellular carcinoma from mainland China.

Hepatocellular carcinoma (HCC) samples from mainland China were examined for the presence and state of hepatitis B virus (HBV) DNA sequences. HBV DNA was detected by dot-blot hybridization in 13 of 17 cases of HCC from the Shanghai area and in three of six samples from Hangzhou. The HCC cases from Shanghai were then analyzed in more detail. Fifteen of the 17 patients had serologic evidence of past or present infection with HBV (with inadequate information available for the other two), and the 13 HCC samples positive for HBV DNA all came from serologically positive patients. Southern blot analysis showed that the HBV DNA sequences were always integrated in the HCC high-molecular-weight DNA; only one or two viral copies were present per tumor cell, and no common integration site was evident. Hybridization analyses using subgenomic probes of HBV DNA revealed that the tumors seldom retained an entire HBV genome. HBV S-region sequences were always present, X-region sequences were usually represented, and C-region sequences were rarely detectable in virus-positive tumors. A fragment within the HBV DNA X-region, between nucleotides 1441 and 1526, was found to hybridize nonspecifically with cellular DNA; reported sequence data indicated that this fragment would contain approximately 70% guanine + cytosine. Histologic sections were prepared from some of the frozen tissue specimens and stained by an indirect immunoperoxidase technique for hepatitis B surface antigen (HBsAg). Only 1 of 10 HBV DNA-positive samples contained HBsAg in the cytoplasm of tumor cells, although abundant HBsAg was present in adjacent normal cells in all 10 cases. There were no significant differences in histology between HCC that contained HBV DNA sequences and those that were virus negative. These data support the premise that HBV represents a major etiologic factor in the development of HCC in the Shanghai area of China, although the molecular basis of viral involvement remains obscure.

Adult↗

PubMed1986

Comparison of risk factors for cervical cancer in different populations.

The incidence of cervical cancer has been found to vary between populations. Risk factors of cervical cancer include early age at first marriage, multiple marriages and antibodies to herpes simplex virus type 2 (HSV-2). The interrelatedness of these risk factors was examined by comparing data collected from 428 cancer cases and 947 control women selected from 6 populations having standardized cervical cancer incidence rates varying from 9.3 to 85.1 per 100,000. Logistic regression analysis revealed that multiple marriages, early age at first marriage or pregnancy and HSV-2 antibodies were all associated with significant risk when all 3 factors were entered into the model. Cervical cancer incidence rates were best predicted by the occurrence of HSV-2 antibodies among control women. To further assess the relation between cervical cancer rates and HSV-2 antibody, 2,306 additional sera representing an 0.8% random sample of females over 9 years of age residing in the Republic of Panama were assayed for antibodies to the virus, and the occurrence of antibodies was correlated with invasive cervical cancer rates specific to each Province. Data from both the random sample and the other study populations yielded a linear relation between the occurrence of HSV-2 antibodies and the incidence of cervical cancer. An exception was found for women living in Herrera Province, Republic of Panama, who had a higher cancer rate than predicted by HSV-2 antibody occurrence. The data suggested that infection with HSV-2 is a co-variable of venereal factors, although a role for the virus in the genesis of a certain proportion of cervical cancers is not excluded.

Adult↗

PubMed1986

Response to a hepatitis B polypeptide vaccine in micelle form in a young adult population.

Polypeptide micelles with relative molecular weights of 25,000 (p25) and 30,000 (gp30) daltons were prepared from native 22-nm hepatitis B surface antigen (HBsAg) particles. This p25/gp30 complex was alum-adsorbed, and three dosage levels (20 micrograms, 4 micrograms, and 0.8 micrograms) were administered at 0, 1, and 6 months to 51 human volunteers. Local and systemic reactions were clinically insignificant, and all vaccinees seroconverted, regardless of dose. As anticipated, antibody responses diminished as the dosage was reduced. Seroconversion rates and geometric mean antibody levels for the 20 micrograms dosage group were significantly better than those observed with a commercial vaccine and were comparable to those achieved after immunization with 40 micrograms of the intact 22-nm particles used to prepare the polypeptides. By 2 weeks, an anti-HBs response was elicited in 80% of the group receiving 20 micrograms of the polypeptide vaccine. This rapid response to immunization may be particularly beneficial for postexposure prophylaxis where the early development of immunity is advantageous.

Adult↗

PubMed1986

Thermostabilization of Inoue-Melnick virus by salt.

Inoue-Melnick virus (IMV) was stabilized to thermal inactivation at 50 degrees C by 1 M Na2SO4 but not by 1 M MgCl2, 1 M MgSO4, 2 M NaCl, and 2 M KCl. The latter salts enhanced the inactivation over that in distilled water. Na2SO4 stabilized IMV at 1 M concentration but enhanced inactivation at 0.1 M. Three types of IMV were stabilized by 1 M Na2SO4 at 50 degrees C to almost the same degree. The pattern of the thermostabilization of IMV by salts was similar to that of herpes simplex virus; however, the degree of its stabilization was lower with prolonged heating.

Herpesviridae↗