[M-29 human diploid cells line--a substrate for the production of antiviral vaccines].

AIM: Study of morphologic and karyologic characteristics of 5 russian human diploid cell lines (HDC). MATERIALS AND METHODS: 5 HDC lines and HDC strain MRC-5 were studied; RK-13 and Vero continuous cell lines were used; viruses: rubella (RA27/3), measles (L-16), epidemic parotitis (L-3). Cytogenetic analysis of HDC was performed by using DAPI differential staining method. RESULTS: M-29 line has characteristics that are similar to those of MRC-5 diploid cell strain. M-29 cell culture is not contaminated with foreign viruses, mycoplasmata, does not have oncogenic potency. CONCLUSION: M-29 line has high virus-productive properties for accumulation of measles, rubella and epidemic parotitis vaccine viruses and may be recommended as a substrate for the production of antiviral vaccines.

[The antiviral action of recombinant forms of the human T-lymphocyte CD4 receptor]. / Protivovirusnoe deistvie rekombinantnykh form CD4-retseptora T-limfotsita cheloveka.

A recombinant protein containing the first 179 N-terminus amino acids of human T-lymphocyte CD4-receptor was synthesized in E. coli cells. This recombinant protein was shown to interact with OKT4A and Leu3a monoclonal antibodies competing with HIV gp120 glycoprotein for binding with the native CD4 receptor. Experiments in vitro in human T-lymphocyte cultures showed that the recombinant CD4-protein in concentrations of 1 to 10 micrograms/ml inhibited the virus-induced syncytium formation, HIV replication in cell culture, synthesis of HIV reverse transcriptase and other virus-specific proteins, that is, behaved as a HIV inhibitor.

[The isolation and characteristics of monoclonal antibodies to recombinant proteins of HIV-1 and HIV-2--the env and gag gene products]. / Poluchenie i kharakteristika monoklonal'nykh antitel k rekombinantnym belkam VICh-1 i VICh-2--produktam genov env i gag.

Ten hybridomas secreting monoclonal antibodies (Mab) against recombinant HIV-1 and HIV-2 antigens were produced (3 Mab anti-gag protein, 2 anti-env1, and 5 anti-env2). In the immunoblotting assay all the anti-gag Mabs reacted with HIV capsid protein p24, whereas one of them reacted also with p55 protein and with 7 other polypeptides. Another anti-gag Mab cross-reacted with the antigen of subpopulation of human peripheral blood lymphocytes. The third one interacted with the antigen of both HIV-1 and HIV-2. All the 10 Mabs interacted with natural HIV antigens and can be used for identification and differentiation of HIV-1 and HIV-2.

[Characteristics of Namalwa cells--a substrate for production of human interferon]. / Kharakteristika kletok Namalwa--substrata dlia polucheniia chelovecheskogo interferona.

The properties of lymphoid Namalwa cell line propagated at the USSR Academy of Medical Sciences Research Institute of Viral Preparations for interferon production are described. The scanning and transmissive electron microscopy studies of the cells showed their morphological stability and the absence of microbial contamination. The 46-48-chromosome cells comprised 85% of the population, hypodiploid cells (44-45 chromosomes), 9%, tetraploid and hypertetraploid cells, 3%. Spontaneous aberrations were detected in 3% of the chromosome. Inoculation of the cells into unsuppressed laboratory animals (rabbits, guinea pigs, adult or suckling mice) or chick embryos did not cause the development of any pathological process. Namalwa cells were shown to produce interferon after multiple (up to 4 times) induction with Newcastle disease virus.

[Isolation of purified and concentrated human interferon from Namalwa cells]. / Poluchenie ochishchennogo i kontsentrirovannogo chelovecheskogo interferona iz kletok liniii Namal'va.

The interferon produced in the cultured Namalwa cells was purified and concentrated according to the method of K. Cantell and S. Hirvonen developed for purification of leukocyte interferon. A preparation concentrated 500-fold had the antiviral activity of 10(6) IU/ml at a specific activity of 1-3 X 10(6) per 1 mg of protein. The amount of protein in the preparation was approximately 0.6 mg/ml, that of cell DNA less than 100 ng/ml. The experiments demonstrated that the virus inducer of interferon and possible contaminant viruses were removed on purification. The preparation had no oncogenic potency on inoculation of newborn Syrian hamsters, was harmless by intraperitoneal inoculation of mice and on multiple inoculations on rabbit eye cornea, was not toxic in cell cultures.

[ELISA immunoenzyme detection of the antigens of interferon and protein contaminants]. / Vyiavlenie antigenov interferona i belkovykh primesei v ego preparatakh s pomoshch'iu immunofermentnogo testa ELISA.

The possibility of specific detection of interferon antigens by ELISA was demonstrated on a model of partially purified and ultrafiltration-concentrated preparations of mouse fibroblast interferon. ELISA allows differentiation of specific interferon antigens and the main protein contaminants of its preparations: bovine serum albumin and virus-inducer proteins.

[Study of LPV oncornavirus reproduction in mixed cell culture]. / Izuchenie reproduktsii onkornavirusa LPV v smeshannoi kul'ture kletok

Electron microscopy and mathematical analysis were used to determine the intensity of LPV oncornavirus production by a single cell in co-cultivated human diploid cells and cells of the continuous T-9 line. Maximum production of intracytoplasmic particles of A type was observed at 48 hours of cultivation, and extracellular virions at 96 hours. Mature virions of B type were more numerous than mature virions of C type in the mixed culture. On the whole the amolnt of mature virions was greater than that of immature ones, and the amount of intracytoplasmic A type particles was considerably greater than that of extracellular particles. By the total amount of production of all virus-specific structures and the two-wave pattern of virus production this mixed culture passaged at a ratio of HDC to T-9 cells 2000 : 1 resembled a culture of T-9 line. Thus, this study indicates that the infected HDC culture at later intervals of cultivation (96 hours) is a more active "producer" of LPV oncornavirus than the main line of T-9 cells or mixed HDC--T-9 culture.

[Clone cells obtained from continuous cultures and differing in their sensitivity to tick-borne encephalitis viruses]. / Issledovanie klonov kletok, poluchennykh iz perevivaemykh kul'tur, otlichaiushchikhsia po chuvstvitel'nosti k virusu kleshchevogo éntsefalita

Studies on cloning of continuous HEp-2 and SPEV cell lines were carried out. The sensitivity of the resulting clones to tick-borne encephalitis virus was determined and the clone lines were shown to be heterologous in their sensitivity to TBE virus by means of the immunofluorescence and virological methods. Among 47 clones of HEp-2 line virus reproduction was observed in 30 clones, no reproduction was demonstrated in 17 clones. The maximum number of cells simultaneously synthesizing virus antigen did not exceed 35% of the population which was conformity with the results obtained in the study of the original HEp-2 cell line. Twenty-nine clones derived from the continuous SPEV cell line were examined. Reproduction of the virus was observed in all of them. However, according to the maximum number of cells involved in the process of antigen synthesis, all the clones could be divided into two groups which differed also by the dynamics of cell involvement in antigen synthesis. The results of the study of clones derived from chronically TBE-infected Hep-2-Soph cell line are presented. In 13 out of 15 clones, the infectious virus and antigen synthesis were demonstrated which suggested that the majority of cells of the parental HEp-2-Soph line had been infected with TBE virus.