Biomedical subjects
Publications and source records attributed to K V Il'in.
A type D retrovirus chronically persisting in HEp-2 cells from human laryngeal carcinoma was analyzed by PCR and sequenced. This virus is most similar to SRV-1 and probably represents one of its subtypes.
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Methods are described that are used for the titration of antinuclear, anticentromere, and anti-Scl-70 antibodies in systemic scleroderma, systemic lupus erythematosus, and rheumatoid arthritis: indirect immunofluorescence with various antigenic substrates (sections of fresh-frozen rat liver and Hep-2 cell culture), counter-current immunoelectrophoresis, isolation of Scl-70 antigen. Use of Hep-2 cells as a substrate for indirect immunofluorescence was found clinically and diagnostically more effective since it permitted the detection of anticentromere antibodies and anti-Scl-70. Nucleolar, mottled, homogeneous, marginal immunofluorescence types were observed when rat liver sections and Hep-2 cells were used for substrates. Anticentromere antibodies and anti-Scl-70 were isolated significantly more frequently in systemic lupus erythematosus or rheumatoid arthritis.
The study of four isolates of chrysanthemum B-virus (CVB) has shown the virus to have a single 40 Kd structural protein able to dissociate under the definite conditions forming the truncated (for 3 and 6 Kd) polypeptides having preserved their whole antigenic determinants. The human plasma is shown to contain antibodies reacting with the structural protein B-BX and, approximately 10 times weaker with the structural protein of another carlavirus, potato M-virus. Interaction of antibodies with CVB is found to take place due to F(ab)2 fragments. The analogous reaction with the proteins of other plant viruses or retroviruses has never been registered. Antibodies reacting with CVB protein are also present in the plasma of green monkey, rabbit, mouse and goat but in lesser quantities than in human plasma. Two possible explanations are proposed for the presented data, either immunization of mammalians by the protein or peptide containing its antigenic determinant, or the accidental coincidence of CVB antigenic determinant with some viral, bacterial or fungal determinant widespread in mammalians.
Screening of antibodies to structural and nonstructural gag gene-coded proteins in humans with lymphadenopathy and AIDS was performed by means of radioimmunoprecipitation (RIP) and western blotting. Pr78gag precursor of gag-coded proteins of type-D retrovirus from Hep-2 cells served as an antigen in RIP tests. Total number of sera (of humans with lymphoadenopathy) under RIP analysis was 18 and one sera of AIDS patient. Six of them reacted with Pr78gag and one out of one AIDS serum. Over 80 sera samples of humans with lymphadenopathy have been tested by means of western blotting with proteins of Mason-Pfizer monkey virus as antigens. Antibodies to p27 (major internal protein of Mason-Pfizer monkey virus) were detected in 12 sera samples of those with lymphadenopathy (dilution 1:100) and in 9 out of 12 sera of AIDS patients (dilution 1:100-1:400). Results obtained make it possible to predict that type-D retroviruses are associated with acquired immunodeficiency syndrome and generalized lymphadenopathy and could play some role in development of this illness in humans.
Sensitivity of protein staining with Serva blue G-250 (Coomassie brilliant blue G-250 analogue) in polyacrylamide gel was determined. It has been shown that protein staining with 0.1% Serva blue G-250 results in the recovery of 80 to 35 ng of single protein, that is almost 10 times higher than reported previously for Coomassie brill. blue G-250 (or R-250) staining. The comparison of the sensitivity of Serva blue G-250 protein staining in PAAG and AgNO3 has shown that AgNO3 staining was approximately 18-30 (but not 100 times, as it had been thought before) times more effective for the majority of proteins under study. Silver staining of some proteins, for instance ribonuclease and a number of retrovirus-specific structural proteins, was of lower efficacy. Thus, to obtain reliable results protein electrophoresis in PAAG should be followed by both staining procedures.
A new gel block for electrophoretic separation is offered. When viewed frontally, the block has a form of a trapezium. During electrophoresis a gradient of voltage is formed in such a block, as a result of which the mobility of macromolecules to be separated markedly decreases. The combination of the block with polyacrylamide gel gradient makes it possible to raise the resolving power of electrophoresis up to 300-400 dalton. The new gel block can be applied successfully to the performance of fine biochemical assays and molecular-biological studies of proteins, glycoproteins and nucleic acids.
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For differentiation of Ilvin-Bykovsky virus (IBV) and monkey Meson-Pfeizer virus (M-PMV) the method of virus neutralization with antibodies against the envelope virus antigen was used. The viruses were cultivated in similar human embryo cells. The results of the virus neutralization were determined by presence or absence of the gs-antigen in the infected cells. The antiserum to M-PMV envelope antigens did not neutralize the IBV antigen. It has been concluded that IBV and M-PMV differ by their envelope antigens and should be regarded as different viruses.
Gs-antigen(s) of the D-type virus isolated from the continuous cells of human carcinoma was detected by the immunodiffusion with an indirect radioimmunoautographic assay in 5 of 7 human breast fibroadenoma. This virus contained gs-antigen common with the MPMV. These data demonstrated the genom of the D-type virus (or of a similar virus) to be integrated with the human fibroadenoma cells genom. Expression of the virus genom takes place at least to the level of the virus gs-antigen synthesis. The D-type virus from the continuous cell cultures of human carcinoma was associated with some human tumours, one of which was fibroadenoma.
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