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Biomedical subjects

M B Gardner

Publications and source records attributed to M B Gardner.

At least 91 records · Page 5Linked to original sources

Rapid, easy, and economical screening test for antibodies to human immunodeficiency virus.

A new dot enzyme immunoassay (EIA) with a conserved portion of the envelope protein of the human immunodeficiency virus (HIV) as antigen has been designed for use in areas with few laboratory facilities and by personnel with little laboratory experience. Sera were tested in 263 subjects who had AIDS or AIDS-related complex or were at-risk or not-at-risk of AIDS from the USA, Africa, and Asia/Oceania. The dot EIA was 100% sensitive in the American subjects, and there were only 2 false negatives in the others, both of which were negative by commercial EIA. The test is simple to perform, economical, rapid (30 min), and stable.

Antibodies, Viral↗

Sequence relationships of type D retroviruses which cause simian acquired immunodeficiency syndrome.

Simian acquired immunodeficiency syndrome (SAIDS) in macaque monkeys is caused by type D retroviruses; three independent virus isolates are identified as SRV-1 (SAIDS retrovirus-serotype 1), SRV-2, and MPMV (Mason-Pfizer monkey virus). Virions from these three isolates have serologically related core antigens, but distinct surface proteins. Also, SRV-2 is unique since it apparently induces retroperitoneal fibromatosis in addition to SAIDS. The complete DNA sequence of molecularly cloned SRV-2 is presented and compared to the sequences of SRV-1 and MPMV and to the sequences of other retroviruses and retroviral-related elements in the genomes of eucaryotic cells. SRV-1 and MPMV show fewer than 6% differences in predicted amino acid sequences encoding gag, prt, pol, and the C-terminal env domain; SRV-2 displays about 15-18% differences in these regions when aligned with SRV-1 or MPMV. Greater variation of predicted amino acid sequences is noted in the externally located N-terminal env domains; SRV-1 and MPMV have 83% homology whereas SRV-2 has 58% homology with either SRV-1 or MPMV. Nucleotide sequences of the LTRs of SRV-1 and MPMV are 88% homologous; SRV-2 shows 70% homology with the LTRs of SRV-1 and MPMV. Comparisons of the predicted pol region amino acid sequences of these simian type D retroviruses with the pol gene of a type B retrovirus, mouse mammary tumor virus (MMTV), reveal about 50% homology. A human endogenous element related to the pol region of MMTV shows about 25% homology of amino acids with the pol sequences of SRV-1 or SRV-2. The prt genes of the simian type D retroviruses are similar in size and predicted amino acid sequence with the prt genes of MMTV and the hamster intracisternal type A particle genome. The C-terminal env domains of the avian type C retrovirus reticuloendotheliosis virus (REV) and the type C baboon endogenous virus (BaEV) have 60 and 85% predicted amino acid homology, respectively, with the C-terminal env domains of SRV-1, SRV-2, and MPMV. Within the gag and pol genes of the simian type D retroviruses there are striking homologies with the rat IgE-binding protein gene. Sequence relatedness of these type D retroviruses with type A, type B, and type C retrovirus genomes and with cellular sequences supports the notion that recombinational events contribute to the genesis and variation of retroviruses.(ABSTRACT TRUNCATED AT 400 WORDS)

Acquired Immunodeficiency Syndrome↗

Comparison of indirect immunofluorescence and Western blot for detection of anti-human immunodeficiency virus antibodies.

There was 100% agreement between the results of indirect immunofluorescence (IF) and Western blot testing when these methods were used to detect antibodies to the human immunodeficiency virus in sera from 25 patients with acquired immune deficiency syndrome (AIDS), 20 patients with AIDS-related complex, 186 subjects at high risk for AIDS, and 40 healthy heterosexuals. However, there was only an 88.7% correlation between IF and Western blot results for 728 sera from blood and plasma donor centers that were selected on the basis of screening enzyme immunoassay reactivity. IF tests yielded nine false-negatives and were equivocal, yielding a nonspecific pattern of reactivity for both infected and uninfected cells for 73 of these specimens. The IF and Western blot methods were equal in performance for the detection of anti-human immunodeficiency virus antibodies in the high-risk and unselected low-risk groups, proving to be a practical approach for testing specimens from these subjects. However, the Western blot was the most acceptable method for the validation of specimens from groups at low risk for AIDS that were selected based on enzyme immunoassay reactivity.

AIDS-Related Complex↗

Induction of simian acquired immune deficiency syndrome (SAIDS) with a molecular clone of a type D SAIDS retrovirus.

We have isolated a molecular clone of the full-length integrated provirus of simian acquired immune deficiency syndrome retrovirus serotype 1 (SRV-1) from a fatal case of simian acquired immune deficiency syndrome in a juvenile rhesus macaque. An integrated SRV-1 provirus was cloned, sequenced, and found to contain four large open reading frames encoding gag-precursor protein, protease, polymerase, and envelope. The proviral clone was transfected into D17 canine osteosarcoma cells and found to produce infectious virus. A comparison of the sequences of this clone with a noninfectious clone showed 20 differences, resulting in 10 amino acid changes. Also, a cluster of exchanges, short insertions, and deletions in the 5' leader sequences resulted in extension of the tRNA(Lys) primer-binding site from 14 to 19 nucleotides. Virus isolated from transfected cells was shown to be infectious and pathogenic, resulting in disease that followed the same time course and mortality as disease induced by uncloned, in vitro cultivated virus isolated from diseased animals. These results unequivocally show that a type D retrovirus (SRV-1) causes a fatal immunosuppressive syndrome in rhesus monkeys.

Acquired Immunodeficiency Syndrome↗

Naturally occurring leukaemia viruses in wild mice: how good a model for humans?

This article presents a historical perspective of the wild mouse leukaemia virus (MuLV) model system in light of the prevailing concepts in the early 1970s when this work was carried out. The epidemiology and natural history of MuLV and its associated lymphoma in wild mice were remarkably predictive of later events associated with human T-lymphotrophic virus infection and adult T-cell leukaemia in man. Striking similarities were noted in the exogenous mode of virus transmission, the regional and familial clustering of virus infection in association with specific lymphomas, the low pathogenicity and long latent period, the neurotropism, the virus stability and the nature of viral integration in resultant lymphomas. The wild mouse model was more accurate than the inbred mouse model in pointing the way to human disease. These results illustrate the importance of studying natural history in the 'real world' and the limitations of laboratory artefacts such as those created by inbreeding of laboratory mice.

Animals↗

Natural history of endemic type D retrovirus infection and acquired immune deficiency syndrome in group-housed rhesus monkeys.

A 2.5-year epidemiologic study of a breeding group of rhesus monkeys (Macaca mulatta), which is a focus of endemic simian acquired immunodeficiency syndrome (SAIDS), demonstrated a strong association between the occurrence of SAIDS and infection with a type D retrovirus, SAIDS retrovirus serotype 1 (SRV-1). Of 23 healthy "tracer" juvenile rhesus monkeys, 19 (83%) died with SAIDS within 9 months of introduction into the resident SAIDS-endemic population. In contrast, 21 healthy "sentinel" juvenile rhesus monkeys placed in the same outdoor enclosure but denied physical contact with the SAIDS-affected group by a 10-foot-wide "buffer zone" remained free of SRV-1, SRV-1 antibody, and disease for 2.5 years. The SAIDS-specific mortality rate was significantly higher in juveniles than in adults. In repeated serologic testing, the overall prevalence of SRV-1 antibody ranged from 68 to 85%. Antibody prevalence increased with age. Seroconversion was found to be a poor indicator of infection rate, as approximately 50% of virus-positive juvenile monkeys had no antibody detectable by enzyme-linked immunosorbent assay. Repeated viral isolations from all animals revealed 1) SRV-1 viremia with clinical SAIDS; 2) persistent viremia and viral shedding in apparently healthy animals; 3) transient viremia and clinical recovery; 4) intermittent viremia, suggesting activation of latent infections; and 5) viremia in a 1-day-old infant, suggesting transplacental transmission. The prevalence of SRV-1 antibody in SAIDS-free breeding groups of rhesus monkeys was 4%. The seroprevalence of antibodies against human T-cell leukemia virus type 1 (HTLV-1), human immunodeficiency virus (HIV), and simian immunodeficiency virus (SIV; formerly STLV-III) was uniformly low or absent in both SAIDS-free and SAIDS-affected groups of rhesus monkeys, demonstrating that these retroviruses are not etiologically linked to SAIDS at the California Primate Research Center.

Acquired Immunodeficiency Syndrome↗

Viremia, antigenemia, and serum antibodies in rhesus macaques infected with simian retrovirus type 1 and their relationship to disease course.

The course of simian retrovirus type 1 (SRV-1) infection was studied in 14 experimentally inoculated juvenile rhesus monkeys. Viral transmembrane protein antigenemia and antibodies to whole virus were measured by enzyme linked immunosorbent assay and correlated with the clinical course of disease and virus isolation. Based on these parameters, animals with simian retrovirus type 1-induced disease were divided into three categories: monkeys dying within a few months of fulminating simian acquired immune deficiency syndrome in the face of a high level persistent antigenemia and viremia, and a nondetectable serum antibody response; monkeys that developed a milder form of simian acquired immune deficiency syndrome but remained alive in spite of a chronic low-grade antigenemia and viremia and only a transient initial antibody response; and monkeys that never became ill and that were either transiently or nontransiently viremic and antigenemic. This latter group developed high levels of serum antibodies. The outcome of simian retrovirus type 1-induced disease was similar to that described for feline leukemia virus infection of cats, another retroviral disease of animals. The disease course differed considerably, however, from that reported for retrovirus-induced human acquired immune deficiency syndrome.

Acquired Immunodeficiency Syndrome↗

Seroepidemiologic survey of captive Old-World primates for antibodies to human and simian retroviruses, and isolation of a lentivirus from sooty mangabeys (Cercocebus atys).

Sera from 526 Old-World monkeys and apes, representing 50 species and 20 genera and living in US zoos and vivaria, were screened for antibodies to HTLV-I, HTLV-III/LAV, and simian-AIDS retrovirus, type I (SRV-I). Sera were screened initially by ELISA, and ELISA-positive sera, as well as ELISA-negative sera from cage contacts, were further tested by Western blotting. A large number of false-positive and a small number of false-negative ELISA sera were identified. Although most true positive reactions were directed to a single retrovirus, a number of individuals from 4 species were positive for more than one retrovirus. Specific seroreactivity to HTLV-I was found in 39/526 (7%) animals of 15 species. True positive reactions to SRV-I were found in 21/516 (4%) animals, including talapoins and 2 species of macaques. Specific serologic reactions to HTLV-III/LAV were detected in 23/526 (4%) monkeys. Many of the HTLV-III/LAV seropositive animals were from one mixed-species zoo exhibit, containing sooty mangabeys, mandrills, Kolb's guenons, and talapoins. A type D virus was isolated from the blood of 3/10 SRV-I antibody-positive Tonkeana macaques, but from none of 11 seropositive talapoins. A lentivirus was isolated from the blood of 4/7 HTLV-III/LAV seropositive sooty mangabeys, but not from seropositive talapoins in the same exhibit or from 2 seropositive colobus from another zoo. The sooty mangabey lentivirus produced generalized lymphadenopathy, leukopenia, and decreased levels of T4 lymphocytes in 2 experimentally infected rhesus macaques.

Animals↗

Distribution of type D retrovirus sequences in tissues of macaques with simian acquired immune deficiency and retroperitoneal fibromatosis.

Horizontally acquired SAIDS retrovirus type 2 (SRV-2), a type D retrovirus related to the Mason-Pfizer monkey virus, has been associated with the simian acquired immunodeficiency syndrome (SAIDS) including retroperitoneal fibromatosis (RF) in several macaque species at two primate research centers. Virus specific gene sequences are present in lymphoid and RF tissues but not in muscle tissue of diseased macaques or in any tissues of uninfected normal monkeys. Serologic and restriction endonuclease mapping techniques have defined unique SRV-2 strains in the Celebes (SRV-2C) and rhesus (SRV-2R) macaques at the Oregon Regional Primate Center, SRV-2 is related to both MPMV and SAIDS type 1 retroviruses and it has no detectable molecular homology with the human AIDS retroviruses.

Acquired Immunodeficiency Syndrome↗

Nucleotide sequence of SRV-1, a type D simian acquired immune deficiency syndrome retrovirus.

Simian acquired immune deficiency syndrome (SAIDS) in the macaque genus of monkeys at the California Primate Research Center is apparently caused by infection by a type D retrovirus. The complete nucleotide sequence (8173 base pairs) of a molecular clone of the prototype SAIDS virus isolate, SRV-1, reveals a typical retrovirus structure with long terminal repeats (346 base pairs) and open reading frames for the gag (663 codons), pol (867 codons), and env (605 codons) genes. SRV-1 also has a separate open reading frame of 314 codons between the gag and pol genes that defines the viral protease gene (prt) and a short open reading frame of unknown significance downstream from the env gene. The SRV-1 protease region shows a high degree of homology to its counterpart in the hamster intracisternal A-type particle genome; both these protease genes are about twice as long as the analogous region of other retroviruses. SRV-1 has no notable similarity in either genetic organization or sequence to the human AIDS retroviruses.

Acquired Immunodeficiency Syndrome↗

Recombinant DNA approaches to feline leukemia virus immunization.

We have utilized 2 recombinant DNA strategies for immunization against FeLV in cats: (a) modified live virus was attenuated by mutation and recombination, and (b) an immunogen, consisting of subunit envelope protein, was prepared in genetically engineered yeast. Results indicated that the genetically manipulated live virus preparations were not protective against FeLV challenge because they were either not attenuated in virulence or were not sufficiently antigenic. Immunization with yeast-synthesized FeLV envelope protein followed by the modified live virus gave protective immunity in cats under experimental conditions. Future immunization attempts will concentrate on enhancing the immunogenic potency of the yeast- synthesized FeLV envelope protein.

Animals↗

The causes of false-positives encountered during the screening of old-world primates for antibodies to human and simian retroviruses by ELISA.

Sera from 526 old-world primates representing 50 different species were screened by ELISA for antibodies to human T-lymphotropic viruses I and III, and simian retrovirus type 1 (SRV-1). About one-fourth of the sera were positive by ELISA. There was a tendency, however, for the same sera to be positive for all three human and simian retroviruses. Only about one in five of the ELISA antibody-positive sera were confirmed to be positive by Western blotting. False-positive ELISA antibody tests were particularly common among sera from mandrills, crab-eating macaques, lion-tailed macaques, African green monkeys, and DeBrazza's and moustached guenons. Sera that were falsely positive in ELISA antibody tests to the three human and simian retroviruses were found to contain antibodies that reacted at comparable intensity with feline leukemia, infectious peritonitis and panleukopenia viruses. The false anti-viral activity of these sera was found to be due to antibodies that reacted with non-viral proteins that were copurified with all five virus preparations. These proteins were present in normal cat and human cells used to grow the various viruses and in gelatin. The implications of nonspecific cell-protein antibodies in primate sera were discussed in the light of this and previous seroepidemiologic studies of man and old-world monkeys.

Animals↗

Human alpha- and beta-interferon but not gamma- suppress the in vitro replication of LAV, HTLV-III, and ARV-2.

The effect of human interferons (IFNs) (alpha, beta, and gamma) on the in vitro replication of AIDS viruses (LAV, HTLV-III, and ARV-2) in human peripheral blood lymphocytes was investigated. At the time of peak virus production, IFN-alpha preparations (leukocyte, Namalwa, alpha 1, and alpha 2) at 100 U/ml, suppressed LAV, HTLV-III, and ARV-2 replication as measured by reverse transcriptase (RT) activity by greater than 50%. This suppression was dose dependent and high dosages (500 U/ml) of IFN-alpha resulted in almost complete suppression of RT activities (77-99%). A low dose (100 U/ml) of IFN-beta suppressed all three AIDS viruses by 75%. In contrast, human IFN-gamma at a dose range from 100 U/ml to 500 U/ml had no significant effect on the production of infectious viruses. These results indicate that only IFN-alpha and -beta are effective against LAV, HTLV-III, and ARV-2 replication. A continuous supply of IFN appeared to be essential for the constant suppression of RT activity. In fact, upon termination of single IFN treatment, enhanced virus production resulted.

Acquired Immunodeficiency Syndrome↗

Evaluation of three commercial screening tests for AIDS virus antibodies.

Three commercial enzyme-linked immunosorbent assays (ELISA) for acquired immune deficiency syndrome (AIDS) virus antibodies were evaluated using serum that had been characterized by an ELISA and western blot procedure developed at the University of California at Davis (UCD). Each of the commercial tests was more specific than the UCD ELISA, but the UCD ELISA was more sensitive in the detection of sera that lacked reactivity by western blot to the envelope glycoprotein (gp-41). The HTLV-III Bio-EnzaBead (Litton Bionetics, Charleston, SC) was less sensitive and specific than the Abbott HTLV-III EIA (Abbott Laboratories, North Chicago, IL) or the Virgo HTLV-III ELISA (Electro-Nucleonics Inc., Columbia, MD). Overall, 22.9% (57 of 250) and 51.0% of sera that were repeatedly (X2) positive by commercial screening kits tested at blood donor centers and clinical laboratories, respectively, were confirmed by western blot. These results indicate that the screening assays for AIDS virus antibodies are not equal in performance and that positive screening test results must be confirmed by a more specific test like western blot before results are released.

Acquired Immunodeficiency Syndrome↗

Transfer, by selective breeding, of the pathogenic Mtv-2 endogenous provirus from the GR strain to a wild mouse line free of endogenous and exogenous mouse mammary tumor virus.

The GR laboratory mouse strain has five endogenous proviral copies of the mouse mammary tumor virus (MMTV). One of these, Mtv-2, is unique because it causes mammary carcinomas in virtually 100% of breeding GR females prior to 1 year of age. To facilitate studies of this locus in particular, and mammary tumorigenesis in general, we genetically tailored a new mouse line, WXG-2, which bears Mtv-2 as its only endogenous MMTV provirus. The WXG-2 line was constructed by making hybrids between the GR strain and a wild mouse line free of both endogenous and exogenous MMTV, backcrossing to the MMTV-free line, and fixing the Mtv-2 locus in a population with the desired genotype. Mammary tumors were observed in 5 of the 20 hybrid females carrying the endogenous Mtv-2 provirus. The WXG-2 line represents a new model system for studying MMTV-induced mammary tumorigenesis in the absence of multiple endogenous proviruses.

Animals↗