Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0002871 (anemia)
 52,094 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A simple method for preparation of highly specific antisera against equine infectious anemia virus proteins p26 and p16 is described. Viral proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The motility of the viral proteins in the gel was compared with standards. Unstained portions of the slab gel were sliced into 5-mm bands, emulsified with Freund's complete adjuvant, and injected into rabbits to produce specific antisera.
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PMID:Simple method for preparation of specific antisera against viral proteins: rabbit antisera against equine infectious anemia virus proteins p26 and p16. 21 92

A 620-bp Bg/II restriction fragment containing the putative protease coding sequence from equine infectious anemia virus (EIAV) proviral DNA was cloned and expressed in E. coli as a Pol precursor protein. In contrast to the 25-kDa fusion protein predicted from the expressed pol sequence, a protein of approximately 10 kDa was generated by apparent autocatalytic processing of the Pol precursor. This mature processed protein was detected in transformed cells using an antisera raised against synthetic peptide from the conserved carboxyl-terminal segment of the predicted EIAV protease coding sequence. Coexpression of this protein with a 35-kDa EIAV Gag-precursor fusion protein resulted in the specific proteolytic processing of the precursor as shown by formation of p26, the major capsid protein of EIAV.
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PMID:Expression of the protease gene of equine infectious anemia virus in Escherichia coli: formation of the mature processed enzyme and specific cleavage of the gag precursor. 131 66

Serological diagnosis of equine infectious anemia is of necessity group-reactive, i.e. based on viral core protein p26, because viral envelope components as well as the host's immune response to them undergo rapid antigenic change. Since 1970 the agar gel-immunodiffusion test ("Coggins-test") has been the diagnostic method of choice. Recently, ELISA tests have been introduced for faster and theoretically more sensitive serodiagnosis, while Western blots have been used to clarify doubtful results obtained in Coggins-tests. A commercial competitive ELISA was found to give practically equivalent results to the Coggins-test. The sensitivity of this market product is intentionally kept marginal in order to avoid false-positive "reactor horses". Another commercial ELISA, non-competitive, gave inconsistent results, creating great turmoil among horse owners when falsely positive. Caution is also indicated when interpreting Western blots. Sera of strongly positive horses gave as many as eleven bands, of medium positives fewer bands, and of the weakest reactors solely the p26 band. Single p26 banding was, however, also encountered in 5% healthy horses, in two of them consistently over time, which are accordingly considered non-specific. In order to be interpreted as positive, a Western blot for this equine lentivirus must band with its core protein plus at least one glycoprotein, similar to the recommended criterion for a positive reading of serum samples from AIDS patients.
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PMID:Equine lentivirus, comparative studies on four serological tests for the diagnosis of equine infectious anaemia. 133 47

Camptothecin (CPT), a topoisomerase I-specific inhibitor, was found in this study to inhibit the replication of equine infectious anemia virus (EIAV) in chronically infected CF2Th cells (designated CF2Th/EIAV). By measuring viral reverse transcriptase activity in the culture medium, we demonstrated that treatment for 1 h with noncytotoxic doses of this drug inhibited production by 32 to 52%, whereas continuous exposure to this drug resulted in an 85 to 92% inhibition. No effect on the viability or growth rate of the cells was detected in any of these treatments. Indirect immunofluorescence analysis of the CPT-treated CF2Th/EIAV cells with anti-p26 capsid protein antibodies showed 60 to 85% reduction in the immunofluorescence-positive cells following drug treatment, and radioimmunoprecipitation analysis of these cells showed a comparable decrease of the pr55gag precursor protein. These data suggest that CPT acts as an anti-EIAV agent to block virus replication in the chronically infected cells.
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PMID:The topoisomerase I inhibitor, camptothecin, inhibits equine infectious anemia virus replication in chronically infected CF2Th cells. 164 21

The uses and limitations of the western blot (WB) and radioimmunoprecipitation assay (RIPA) techniques for study of feline immunodeficiency virus (FIV) and FeLV were evaluated. Western blot analysis was used to detect antigenic relatedness between the 2 lentiviruses. Using a rabbit serum directed against p26 of the equine infectious anemia virus (EIAV) and anti-EIAV horse serum obtained from an infected horse, cross-reactivity with p24 of FIV was revealed. Cat sera obtained late after experimentally induced FIV infection recognized p26 of EIAV, which indicates reciprocal cross-reactivity. For RIPA, FIV was metabolically labeled, and virus-coded proteins were identified, using immunoprecipitation. Polypeptides with apparent molecular mass of about 15, 24, 43, 50, 120, and 160 kilodaltons were detected. An additional polypeptide of 10 kilodaltons was found only by use of WB analysis.
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PMID:Use of western blot and radioimmunoprecipitation for diagnosis of feline leukemia and feline immunodeficiency virus infections. 166 78

Feline immunodeficiency virus (FIV) grown in cat lymphocyte and thymocyte cultures was labelled with L-[35S]methionine or [3H]glucosamine and virus-coded proteins were identified using immunoprecipitation. Polypeptides with apparent Mr values of 15K, 24K, 43K, 50K, 120K and 160K were detected. An additional polypeptide of 10K was detected by Western blot analysis. The two highest Mr species sometimes appeared as one band, of which only the 120K polypeptide was glycosylated. In the presence of tunicamycin gp120 was no longer detectable and a non-glycosylated precursor of 75K was found instead. Pulse-chase experiments suggested that the smaller polypeptides p24 and p15 are cleavage products of both p160 and p50. Western blot analysis using a rabbit serum directed against p26 of equine infectious anaemia virus (EIAV) and an anti-EIAV horse serum from a field case of infection revealed a cross-reactivity with p24 of FIV. Cat sera collected late after experimental FIV infection recognized p26 of EIAV, indicating a reciprocal cross-reactivity.
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PMID:Intracellular proteins of feline immunodeficiency virus and their antigenic relationship with equine infectious anaemia virus proteins. 169 Feb 64

A panel of recombinant trpLE-gag fusion proteins and synthetic peptides was used in Western immunoblot and enzyme-linked immunosorbent assays to identify segments of the major core protein (p26) of equine infectious anemia virus that are antigenic in horses during experimental and natural infections with the virus. The predominant humoral immune response was directed toward a highly immunogenic domain composed of 83 amino acids from the carboxy terminus of p26. The observed immunogenicity of p26 resembled that reported for p24 of human immunodeficiency virus type 1, suggesting the conservation of structural motifs in the lentiviral core proteins which are responsible for their observed immunogenicity during persistent lentivirus infections.
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PMID:Characterization of the antigenic domains of the major core protein (p26) of equine infectious anemia virus. 170 39

Three ponies were inoculated with plasma containing 10(4.8) TCID50 of equine infectious anemia virus (EIAV) and observed for 165 to 440 days. Each pony developed a febrile response within 3 weeks of infection during which a plasma viremia greater than or equal to 10(3.5) TCID50/ml was observed. Analyses of four isolates from sequential febrile episodes in a single pony were conducted by two-dimensional tryptic peptide maps and with monoclonal antibodies in immunoblots. Structural and antigenic alterations were observed in the envelope glycoproteins gp90 and gp45, with greatest variation in gp90. Specific IgG to EIAV gp90, gp45, and p26 of homologous and heterologous isolates was detectable by immunoblots within one month after infection although IgG levels to gp45 at this time were relatively low. The group-specific determinants of gp90 and gp45 were more antigenic than those of p26; however, binding of IgG to these determinants did not correlate with neutralization of EIAV as assayed in fetal equine kidney cells. Neutralizing antibodies were first detectable within two months of infection and only neutralized viruses isolated prior to serum collection. Neutralizing activity of sera collected later in the infection was broadly reactive regardless of the number of clinical episodes the donor had suffered.
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PMID:Equine infectious anemia virus (EIAV) humoral responses of recipient ponies and antigenic variation during persistent infection. 216 60

We describe a staining technique, using Ponceau S in very mild conditions, by which proteins can be visualized on nitrocellulose replicas without being permanently fixed to the membrane itself, thus allowing subsequent procedures such as immunoblotting or preparative elution of the proteins to be performed. This staining technique can detect 250 to 500 ng protein, which is essentially the same sensitivity seen for Coomassie blue staining of proteins on nitrocellulose. The Ponceau S staining technique was used to locate proteins on nitrocellulose replicas for subsequent in situ radioiodination and trypsin digestion, followed by separation of the resultant digests in two-dimensional peptide analysis. Staining proteins with Ponceau S did not interfere with either the radioiodination or trypsin digestion, as indicated by essentially identical peptide patterns being obtained for the internal protein p26 from equine infectious anemia virus, regardless of whether the digests were prepared from polyacrylamide gel slices or nitrocellulose sections. The combination of preparation of radioiodinated tryptic digests on nitrocellulose and subsequent two-dimensional analysis is sensitive enough to detect peptide additions and deletions occurring in the surface antigen gp90 recovered from two antigenically distinct strains of equine infectious anemia virus. Thus these procedures provide a relatively simple, inexpensive, and highly reproducible technique for the analysis of as little as 250 nanograms of protein after separation by electrophoresis in polyacrylamide gels.
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PMID:Reversible staining and peptide mapping of proteins transferred to nitrocellulose after separation by sodium dodecylsulfate-polyacrylamide gel electrophoresis. 242 81

Capsids of equine infectious anemia virus have been isolated as cone-shaped particles 60 x 120 nm in size. Detergent treatment of whole virus followed by two cycles of rate-zonal centrifugation in Ficoll produces these capsids in a yield of approximately 10%. The major protein components are the gag-encoded p11 nucleocapsid protein and p26 capsid protein, which are present in equimolar amounts. Substantial cleavage of p11 to p6 and p4 can be observed under conditions where the viral protease packaged in the capsid is enzymatically active.
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PMID:The preparation and biochemical characterization of intact capsids of equine infectious anemia virus. 254 3


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