Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
 31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The C-type particles observed by electron microscopy in PK-15 cells were demonstrated to have biochemical and biophysical properties associated with the oncornavirus group: density of 1:16 in a sucrose gradient, 70S RNA, and the RNA-dependent DNA polymerase. The group-specific interspecies antigen, gs-3, was not present. Evidence of a latent infection with a porcine parvovirus was also obtained.
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PMID:Antigenic and biochemical characterization of the C-type particle of the stable porcine kidney cell line PK-15. 412 28

The nonstructural protein-2 (NS2) of canine parvovirus (CPV) is produced from the left-hand open reading frame of the viral genome and contains 87 amino-terminal amino acids in common with nonstructural protein 1 (NS1) joined to 78 amino acids from an alternative open reading frame. In the minute virus of mice parvovirus NS2 plays a role in controlling capsid protein assembly and translation in a host-specific manner. The predicted NS2 of CPV is divergent from the proteins of the rodent parvoviruses, and the protein and its functions have not been described. We characterized the large and the small splices of CPV using reverse transcriptase-PCR, NS2 was identified using anti-peptide antibodies against the predicted C-terminal sequence and also by expressing the protein from a plasmid vector. The protein could be detected at low levels in the nucleus and the cytoplasm of a proportion of CPV-infected cells, as well as in cells transfected with the expression plasmid. Virus genomes were prepared with mutations in the splice donor or acceptor sites of the NS2-specific intron or with three different termination codons in the NS2-unique exon. Both splice donor and acceptor mutations resulted in the use of previously cryptic splice sites, and the virus containing the splice donor mutation replicated inefficiently. However, the other four mutant viruses were all viable and replicated efficiently in cat and dog cells, and two mutant viruses that were tested appeared to assemble their capsids in the same manner as did the wildtype. After inoculation of dogs an NS2 mutant virus with a termination codon in the NS2-unique exon replicated to titers similar to those seen for wildtype CPV in several tissues examined.
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PMID:Nonstructural protein-2 and the replication of canine parvovirus. 945 1

Infection with human parvovirus B19 causes fifth disease, acute and chronic red cell aplasia, fetal hydrops, arthropathy, and other disorders. Antiviral antibodies limit B19 infection in vivo; however, the identification of serologic markers of protection has been hampered by the lack of a quantitative assay for parvovirus neutralization. A novel in vitro test for parvovirus neutralization has been developed using reverse transcriptase-polymerase chain reaction to detect viral transcripts in a B19-permissive cell line. Parvovirus neutralizing activity was measured in sera from naturally infected individuals, and common features of sera with high neutralizing capacity were identified as protection correlates. Sera that suppressed B19 replication in vitro demonstrated IgG reactivity with capsid proteins VP1 and VP2, but no linear relationship between antibody titer and neutralizing capacity was observed. Sera from experimental animals and human volunteers immunized with a virus-like particle vaccine candidate exhibited B19 neutralizing titers equal to or greater than those observed in natural infections.
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PMID:Quantitative analysis of neutralizing immune responses to human parvovirus B19 using a novel reverse transcriptase-polymerase chain reaction-based assay. 995 68

Fecal samples were examined for viruses participated in gastrointestinal disorders of cats, especially focusing on feline coronavirus (FCoV) by a reverse transcriptase-polymerase chain reaction assay. It was found that a primary viral pathogen was feline panleukopenia parvovirus (FPLV; 28.5% of the positive rate) and the secondary was FCoV (10.7%). Commonly reported clinical signs of cats of which feces were FCoV-positive were vomiting, diarrhea and dehydration with an exception of one serious case with concurrent FPLV infection.
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PMID:Feline coronavirus participation in diarrhea of cats. 1053 17

It has long been recognized that skin may be a particularly good target for pharmacologic gene therapy and as a platform for the secretion of systemically distributed molecules. Adeno-associated virus type 2 (AAV) is a useful vector for skin gene therapy because skin is the natural host tissue for AAV, in which it functions as an autonomous parvovirus. We demonstrate here that recombinant (r) AAV vectors carrying the granulocyte-macrophage colony-stimulating factor (GM-CSF), human papillomavirus E6, or green fluorescent protein (GFP) transgene could transduce primary human keratinocytes in ex vivo culture. We further demonstrate that these transduced cells could be used to form a transgene-positive recombinant skin (r-skin), using the organotypic epithelial raft culture system. Transduction of keratinocytes was demonstrated by reverse transcriptase-polymerase chain reaction (RT-PCR) for RNA expression, enzyme-linked immunosorbent assay (ELISA) for product secretion, intracellular staining for protein expression, vector-chromosomal junction PCR and Southern blot analysis of proviral sequences, in situ immunohistochemistry analysis of protein expression, and ultraviolet light fluorescence for GFP expression. AAV/GM-CSF/Neo-infected keratinocyte/raft skins secreted GM-CSF at levels as high as 25 ng/cm(2) of skin and maintained expression to 60 days postinfection. These data support the utility and efficiency of AAV-based gene delivery to produce genetically altered keratinocytes and r-skin.
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PMID:Generation of recombinant skin in vitro by adeno-associated virus type 2 vector transduction. 1568 79

Although IgE is implicated in viral immunity, its role in parvovirus B19 immunity and its relationship to other immunological states has not been studied. Total serum immunoglobulin levels, IgG and IgE anti-parvovirus B19, blood lymphocyte numbers, and epsilon and cytokine specific mRNA were determined in pediatric patients with normal serum IgA levels (IgA+) and selective IgA deficiency (IgA-) on days 0 (initial diagnosis) and 14, and 3 years after recovery (nephelometry, Western blot test, flow cytometry, reverse transcriptase-polymerase chain reaction). We found that both patients had serum IgM, IgG, IgE, and IgA levels within normal ranges on day 0 to 3 years, excluding IgG(1) and IgA in the IgA- patient, which were elevated and negative, respectively, and IgE in the IgA+ patient, which was elevated (>100 IU/ml). The serum IgA+ and IgA- patients made IgE (and IgG) anti-parvovirus B19 at all time points. Excluding CD8(+)CD60+ T cells, determinations of T, B, and NK lymphocyte subsets always were within normal ranges. In both patients, CD8(+)CD60+ T-cell numbers were within normal ranges on day 0, but dramatically increased on day 14 (more than fivefold). At 3 years, they had returned to normal in the IgA+ patient, but remained high in the IgA- patient. On day 0 to 3 years, peripheral blood mononuclear cells of both patients expressed epsilon- and interferon (IFN)-alpha-specific mRNA. On day 0, the IgA+ patient expressed interleukin (IL)-4 and IL-10, but not IL-2, IFN-gamma, or IL-6 mRNA; the IgA- patient expressed IL-6 and IL-10 mRNA, but not IL-4, IL-2, or IFN-gamma mRNA. At 3 years, the IgA+ patient expressed mRNA for all cytokines, but the IgA- patient did not express mRNA for any of these cytokines. Our results suggest that IgE is important in parvovirus B19 immunity, and that IFN-alpha and CD8(+)CD60+ T cells may regulate IgE memory responses and isotype switching.
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PMID:IgE, CD8(+)CD60+ T cells and IFN-alpha in human immunity to parvovirus B19 in selective IgA deficiency. 1638 44

Parvovirus B19 is a single-stranded DNA virus exhibiting affinity for a variety of cell types including endothelial cells. The basis of the affinity is globoside expression, the receptor for B19. B19 endothelial cell parasitism accelerates endothelial cell apoptosis, potentially critical in inducing neoantigenicity and secondary antibody formation. A 38-year-old previously healthy male developed intestinal and cutaneous infarction, elevated creatinine phosphokinase (CPK) levels and progressive cytopenias. The basis of his clinical presentation was unclear until a skin biopsy suggested a potential role for virally mediated endothelial cell injury. Hematoxylin and eosin, immunohistochemical, immunofluorescent and reverse transcriptase in situ polymerase chain reaction studies were conducted on skin biopsies to assess for viral triggers. B19 viremia and localization of B19 RNA transcripts to vascular endothelium were uncovered. A diagnosis of catastrophic endothelial cell injury attributable to B19 infection was rendered. The patient showed significant improvement with intravenous immunoglobulin (IVIG). Parvovirus B19 is a ubiquitous virus that in the majority of affected patients remains asymptomatic. Nevertheless, because of its ability to infect endothelium it can cause severe multiorgan endothelial cell injury.
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PMID:Parvovirus B19-associated catastrophic endothelialitis with a Degos-like presentation. 1853 56

We developed a reverse transcriptase polymerase chain reaction (RT-PCR) method for the detection of duck hepatitis virus type 1 (DHV-1) in the tissues of infected and clinically affected ducks and in chick and duck embryos. We found the assay to be effective in detecting the virus in China, where it is being used in studies on the epidemiology of the disease. We applied this simple and rapid diagnostic method to the detection of DHV isolates grown in chick and duck embryos and in tissues obtained from infected birds. The assay also proved useful for the differentiation of DVH from the duck plague virus (DPV), muscovy parvovirus (MPV), gosling parvovirus (GPV), avian influenza virus (AIV/H5N1), Pasteurella multocida (PA/5:A), Riemerella anatipestifer (RA/serotype 1), and Salmonella enteritidis (SE). The limit of the sensitivity of this method for the detection of DHV-1 RNA was 3 pg/10 microl. As compared to ELISA and virus isolation, the rate of agreement for the detection of experimentally infected livers was 100%; moreover, the RT-PCR method was also capable of detecting DHV-1 RNA from the livers that had been infected and stored at -20 degrees C for 22 years; in contrast, ELISA and virus isolation method could only detect DHV-1 from the livers that had been infected and stored at -20 degrees C for 13 and 11 years, respectively. The rate of positivity in 185 clinically suspected diseased livers subjected to detection by RT-PCR, ELISA, and virus isolation was 89.2%, 69.2%, and 55.7%, respectively. These results indicated that the RT-PCR approach is rapid, sensitive, and reliable for the detection and differentiation of DHV-1 from the other clinical samples and suspected isolates.
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PMID:Development and application of a reverse transcriptase polymerase chain reaction to detect Chinese isolates of duck hepatitis virus type 1. 1947 29

We developed a reverse transcriptase polymerase chain reaction (RT-PCR) method for the detection of duck hepatitis virus type 1 (DHV-1) in the tissues of infected and clinically affected ducks and in chick and duck embryos. We found the assay to be effective in detecting the virus in China, where it is being used in studies on the epidemiology of the disease. We applied this simple and rapid diagnostic method to the detection of DHV isolates grown in chick and duck embryos and in tissues obtained from infected birds. The assay also proved useful for the differentiation of DVH from the duck plague virus (DPV), muscovy parvovirus (MPV), gosling parvovirus (GPV), avian influenza virus (AIV/H5N1), Pasteurella multocida (PA/5:A), Riemerella anatipestifer (RA/serotype 1), and Salmonella enteritidis (SE). The limit of the sensitivity of this method for the detection of DHV-1 RNA was 3 pg/10 microl. As compared to Dot-ELISA and virus isolation, the rate of agreement for the detection of experimentally infected livers was 100%; moreover, the RT-PCR method was also capable of detecting DHV-1 RNA from the livers that had been infected and stored at -20 degrees C for 22 years; in contrast, Dot-ELISA and virus isolation method could only detect DHV-1 from the livers that had been infected and stored at -20 degrees C for 13 and 11 years, respectively. The rate of positivity in 185 clinically suspected diseased livers subjected to detection by RT-PCR, Dot-ELISA, and virus isolation was 89.2%, 69.2%, and 55.7%, respectively. These results indicated that the RT-PCR approach is rapid, sensitive, and reliable for the detection and differentiation of DHV-1 from the other clinical samples and suspected isolates.
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PMID:Development and application of a reverse transcriptase polymerase chain reaction to detect Chinese isolates of duck hepatitis virus type 1. 1870 44

To evaluate the effects of recombinant porcine interleukin-18 (rpIL-18) on the replication of viruses in host cells and proliferation of lymphocytes, porcine IL-18 (pIL-18) isolated from a domestic big-white porcine breed found in the Henan province (HN) was cloned using a reverse transcriptase-PCR. The cloned HN pIL-18 contained an ORF of 579 base pairs encoding a 192-amino-acid precursor protein. The amino acid sequence of HN pIL-18 was compared with all the other pIL-18 amino acid sequences and varied by at least one amino acid to the consensus of all the others available. HN pIL-18 mature protein gene was inserted into a prokaryotic vector pGEX-4T-1 and expressed in Escherichia coli BL21. The expression of glutathione-S-transferase-pIL18 fusion protein was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis. The rpIL-18 induced in vitro proliferation of concanavalin-A-stimulated porcine splenocytes, as revealed by the MTT assay. We studied the antiviral activities of the rpIL-18 on the replication of porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), and porcine parvovirus (PPV) cultured in two homologous cell lines. The results suggested that rpIL-18 can stimulate the proliferation of lymphocytes and inhibit viral pathogens infecting the porcine population.
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PMID:Cloning, in vitro expression, and bioactivity of interleukin-18 isolated from a domestic porcine breed found in Henan. 1973 42


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