[The study of experimental and natural rotavirus infection in mice using a heterologous enzyme immunoassay test-system]. / Izuchenie éksperimental'noi i estestvennoi rotavirusnoi infektsii myshei s pomoshch'iu geterologichnykh test-sistem immunofermentnogo analiza.

Enzyme immunoassay test systems constructed on the basis of heterologous calf and monkey rotaviruses were shown to be applicable for diagnosis and study of the pathogenesis of experimental and natural rotavirus infection of mice.

[Development of test systems of immunoenzyme analysis for the rapid diagnosis of rotavirus infection]. / Razrabotka test-sistem immunofermentnogo analiza dlia bystroi diagnostiki rotavirusnoi infektsii.

A test system of enzyme immunoassay (EIA) consisting of simian rotavirus SA-11 and rabbit antiserum has been developed for the detection of rotavirus antigen. Direct EIA was used for tests on stool specimens from 289 children varying in ages from 10 days to 12 years suffering from acute enteric infections and 56 normal children. The antigen was detected in 22.1% and 3.5%, respectively, in patients predominantly in the first 3 days of the disease and most frequently in cases running as gastroenteritis. The results are discussed with reference to the possibility of the development of rotavirus carrier state and mixed virus-bacteria infections.

[Suppression of rotavirus SA-11 reproduction by protease inhibitors in cell culture]. / Podavlenie reproduktsii rotavirusa SA-11 ingibitorami proteaz v kul'ture kletok.

The effect of proteases inhibitors, epsilon-amino-caproic acid and gordox, on reproduction of rotavirus SA-11 in MA-104 cells was studied by enzyme immunoassay. These inhibitors were shown to exert an inhibiting effect on rotavirus reproduction.

[Scanning microscopy of hamster cells transformed by herpes simplex virus type 2]. / Rastrovaia mikroskopiia kletok khomiachka, transformirovannykh virusom prostogo gerpesa tipa 2.

Scanning microscopy was used to examine the features of morphology and attachment to a solid substrate of a transformed and tumor lines of hamster cells. These cell lines differed from normal hamster fibroblasts by changes in the mode of attachment and the degree of flattening on the solid substrate, relief of the cell surface and pattern of intercellular interactions. The observed morphological changes correlated with the degree of cell transformation.

[Mechanism of the cellular transformation of newborn hamsters inoculated with herpes simplex type 2 virus]. / Izuchenie mekhanizma transformatsii kletok novorozhdennogo khomiachka, inokulirovannykh virusom prostogo gerpesa tipa 2.

Combined virological, antigenic, electron microscopic, and molecular biologic study of the role of an oncovirus, herpes simplex type 2 virus (HSV-2), in the mechanisms of transformation of HSV-2-infected hamster cells was carried out. No expression of genetic information of the oncovirus could be detected. Cell transformation was shown to be associated with persistence and realization of HSV-2 genetic information in the transformed cells.

[Morphological, cytogenetic and proliferative characteristics of Syrian hamster HTC-2 and HTC-1 cells and of the HTCT tumor cell line transformed by herpes simplex type 2 virus]. / Morfologicheskie, tsitogeneticheskie i proliferativnye osobennosti transformirovannykh virusom prostogo gerpesa tipa 2 kletok siriiskogo khomiachka KhTK-2, KhTK-1 i linii opukholevykh kletok KhTKO.

Morphological, caryological, cytoproliferative, and isoenzyme studies of Syrian hamster cells HTC-2 and HTC-1 transformed by herpes simplex virus type 2 and a HTCT line of tumor cells were carried out. The hamster origin of these cell lines was established by chromosomal analysis and electrophoretic mobility of lactate dehydrogenase and glucose-6-phosphate-dehydrogenase isoenzymes. The transformed and tumor cell lines differ from normal cells by altered morphology, an aneuploid chromosome set and increased proliferative activity. Electron microscopic examination of transformed cells revealed increased amounts of filamentous and tubular structures in the cytoplasm. No retroviruses were found.

[Malignant transformation of newborn hamster cells by herpes simplex virus type 2]. / Zlokachestvennaia transformatsiia kletok novorozhdennykh khomiakov virusom prostogo gerpesa tipa 2.

The oncogenic potentials of HSV-2 inactivated by ultraviolet rays in newborn hamster cell culture were studied. Virus-induced morphological and malignant transformation of cells was accompanied by synthesis of virus-specific antigen, changes in morphology, formation of colonies in semi-liquid agar, and tumorigenicity of cells. The animals bearing tumors induced by transformed cells showed humoral and cellular immune responses to the virus-specific antigen. Lines of transformed and tumor cells were obtained and established in passages.

[Cellular immune response detected in blast transformation test with lymphocytes from rabbits immunized with nucleocapsid of herpes simplex type 2 virus]. / Kletochnyi immunnyi otvet, vyiavliaemyi v reaktsii blasttransformatsii limfotsitov krolikov, immunizirovannykh nukleokapsidom virusa prostogo gerpesa tipa 2.

A preparation of herpes simplex virus type 2 (HSV-2) nucleocapsid (NC) was obtained after treatment of intracellular virus with detergents followed by centrifugation in linear sucrose density gradient. Electron microscopic analysis revealed polymorphism of HSV-2 NC population. Three main types of capsid structures were found: empty capsids, NC containing various amounts of nucleoid material, and NC with complete nucleoid. Morphological heterogeneity correlated with heterogenous sedimentation profile of the total NC preparation of labeled virus represented by light, intermediate, and heavy structures. Specific blasttranformation tests with lymphocytes from rabbits immunized with HSV-2 NC demonstrated differences in the primary and secondary cellular immune responses. The secondary cellular immune response was characterized by a more rapid and strong increase in BTT values than the primary immune response.

[Etiological role of the herpes simplex virus in causing cervical cancer]. / Izuchenie étiologicheskoi roli virusa obychnogo gerpesa v vozniknoveniia raka sheiki matki.

The results of the composite virological and immunological examinations of 46 patients with cervical carcinoma (CC) attest to the association of herpes simplex virus (HSV) with neoplastic processes. HSV strains were isolated from the tumour tissue as well as from the cervical canal secretions and blood of 4 patients. A higher level of virus-neutralizing and complement-fixing antibodies to HSV was found in sera from CC patients as compared with the control group of normal women. Antibody to HSV type 2 was found in 38.8% of sera from the patients and 12.6% in the control group. Lymphocytes from CC patients induced blasttransformation reaction to HSV twice as frequently as those from healthy subjects. The virus-specific antigen was found in tumour cells of 36% CC patients in immunofluorescent examinations of smears from the cervix. The presented results are related to the determination of the etiological role of HSV in cervical carcinoma.

[Modelling of viral-chemical carcinogenesis in chronic infection in mice caused by the herpes simplex type 2 virus]. / Modelirovanie virusno-khimicheskogo kantserogeneza v usloviiakh khronicheskoi infektsii, vyzvannoi virusom prostogo gerpesa tipa 2.

The influence of chronic herpetic infection of white mice caused by herpes simplex virus type 2 (HSV-2) on the blastomogenic effect of a polycyclic hydro-carbon, 20-methylcholanthrene (20-MCA), was studied. Solid tumors developed in 20-MCA-treated mice on the side of the carcinogen administration within 81--89 days. The simultaneous administration of 70 intracerebral LD50 of HSV-2 subcutaneously into a hind leg pad and 0.1 mg 20-MCA subcutaneously into the inguinal area was accompanied by a synergistic action of the two factors. The number of tumors in the animals treated with HSV-2 in combination with 20-MCA at all intervals of observation was 1.5--2.5 times greater than in mice treated with 20-MCA alone. Mice weighing 10--12 g were more sensitive to the synergistic effect of HSV-2 and 20-MCA, whereas animals weighing 25--30 g were more sensitive to the carcinogenic effect of 20-MCA alone. No tumors developed in mice infected with HSV-2 alone by 167 days (the observation period). Mice weighing 10--12 g were found to be more sensitive to the fatal effect of HSV-2 and HSV-2 plus 20-MCA, particularly in the first 2--3 weeks after inoculation. Six months after inoculation with HSV-2 the virus was isolated from posterior root ganglia of the sacral and lumbar spinal cord by cocultivation of the ganglia with human embryo skin-muscle tissue cells.

[Comparative study of the effectiveness of the purification and concentration of herpes simplex virus types 1 and 2]. / Sravnitel'noe izuchenie éffektivnosti ochistki i kontsentratsii virusa prostogo gerpesa tipov 1 i 2.

The results of comparative studies on concentration and purification of herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2) by ficol density gradient centrifugation are presented. A two-phase distribution of extracellular HSV was established in phycol density gradient centrifugation: in zones with density of 1.110-1.114 and 1.088-1.085 g/ml. The effectiveness of purification of HSV preparations recovered from the corresponding gradient zones was determined by electron microscopy and quantitation of the contaminating cellular (radioactive) proteins in virus purification from a mixture of the culture fluid from infected cultures and the culture fluid from uninfected labeled human embryo skin-muscle tissue cultures (HESM) and a mixture of unlabeled extracellular HSV and a homogenate of labeled uninfected HESM cultures. In HSV purification from the virus-containing culture fluid the amount of cellular proteins was shown to decrease 500-fold in 150-fold virus concentration. In purification of extracellular HSV from the mixture with cell homogenate the amount of cellular proteins decreased 70- and 100-fold for HSV-2 and HSV-1, respectively. The infectious virus yield in phycol gradient centrifugation of a precipitate obtained by the addition of polyethylene glycol-6000 to the culture fluid for HSV-1 was 34.7% (in titrations in HESM cultures) and 38.4% (by intracerebral inoculation of mice weighing 5-6 g), and for HSV-2 20.2% and 26.3%, respectively.

[Comparative study of the oncornavirus A and D proteins of human J-96 cells by the methods of isoelectric focusing and polyacrylamide gel electrophoresis]. / Sravnitel'noe izuchenie belkov onkornavirusov A i D v perevivaemykh kletok cheloveka J-96 metodami izoélektricheskogo fokusirovaniia i élektroforeza v poliakrilamidnom gele.

Three peaks of 14C-radioactivity with buoyant densities of 1.23--1.24, 1.26 and 1.29 g/ml were detected in a cytoplasmic extract of J-96 cells upon equilibrium centrifugation in sucrose gradient. Electron microscopy of the 1.23--1.24 g/ml buoyant density fraction revealed particles 60--80 nm in diameter showing morphology characteristic of oncornavirus A. Isoelectric focusing in polyacrylamide gel showed polypeptides of extracellular D virus and oncornavirus A to differ in isofocusing points (pI). Proteins of extracellular D virus were localized in zones with pH 3.7, 4.0, 4.4, 4.7, 5.6, 6.5, 8.1, 9.45, and 10.0; polypeptide of intracytoplasmic oncornavirus A had the following isofocusing points: 4.0, 4.9, 6.7, 7.3, 9.0, 9.45 and over 10.0. Electrophoresis of polypeptides of D virus and intracellular oncornavirus A revealed differences in the molecular weights of the components. No proteins with molecular weights of 10,000, 12,000, 15,000, and 27,000 dalton characteristic of the extracellular D virus were found in oncornavirus A virions. The analysis of protein patterns obtained in parallel experiments of isoelectric focusing and polyacrylamide gel electrophoresis suggests that oncornaviruses A and D of J-96 cells differ in the characteristics (pI and molecular weight) of the structural polypeptide components.

[Protein metabolism in human embryo skin-muscle tissue cells infected with oncornavirus D from J-96 cells]. / Metabolizm belkov v kletkakh kozhno-myshechnoi tkani émbriona cheloveka, infitsirovannykh onkornavirusom D iz kletok J-96.

Structural polypeptides of oncornavirus D produced by J-96 cells and the effect of oncornavirus infection on protein metabolism of human embryo skin-muscle tissue cells (HESM) were studied. Electrophoresis showed the number of polypeptides detectable in a preparation of virus produced by infected HESM cells to depend on the number of centrifugations in sucrose gradient in the process of virus purification. A preparation of a singly purified virus was found to contain 13 polypeptide components: p115, p110, p100, p87, p78, p69, p57, p45, p36, p27, p15, p12, and p10. In oncornavirus D from J-96 cells purified three times in sucrose gradient 2 major polypeptides were found with molecular weights 69,000 and 27,000 daltons which had about 80% of the total radioactivity, and minor polypeptides p15, p12, and p10. Comparison of electrophoregrams of proteins of HEMS cells infected with oncornavirus D and uninfected cells showed oncornavirus infection to be accompanied by an increased synthesis of some polypeptides (p115, p87, p69) and a decreased amount of others (p78, p36, p17-p12). Besides, in infected HESM cells two proteins were found the molecular weights of which coincided with those of oncornavirus polypeptides p69 and p27. The p69 protein component was detected in the membrane fraction of a continuous line of transformed J-96 cells labeled with 3H-glucosamine indicating its complex glycoprotein nature. The results suggest that the observed changes in metabolism of cell proteins in oncornavirus infection of HESM cells do not result in transformation of these cells or are insufficient for its occurrence.

[Effect of exogenous interferon on the course of latent oncornavirus infection of J-96 cells]. / Vliianie ékzogennogo interferona na techenie latentnoi onkornavirusnoi infektsii kletok J-96.

The effect of exogenous leukocyte interferon on the course of chronic oncornavirus infection of lymphoblastoid J-96 cells chronically producing type B virus was studied. By means of radio-isotope analysis and electron microscope examinations it was shown that upon long-term passage of the cells (18-34 passages) in the presence of interferon (10 units/ml) the process of virion formation in the cells was inhibited 2.0-4.4-fold and virus budding to a lower extent (1.6-2.7-fold). Interferon exerted no inhibiting effect on the formation of intracytoplasmic virions of type A. The employment of KC-test showed the oncornavirus produced by J-96 cells in the presence of interferon to have retained its biological activity, being able to induce synthesis of the indicator KC cells. Examination of the cell membranes by electrophoresis in polyacryl amide gel showed that interferon contributed to accumulation of glycoproteins with high molecular weights (115 000, 100 000 and 68 ooo daltons) in this cell fraction. Simultaneously the experimental cells were found to have 2-3-fold increased amount of inter-species group-specific antigen of Mason-Pfizer virus. The mechanism of action of interferon on multiplication of J-96 cell oncornavirus is discussed.

[Assessment of the sensitivity of immunoprecipitation for the serodiagnosis of influenzal-bacterial pneumonias]. / Otsenka chuvstvitel'nosti immunopretsipitatsii dlia serodiagnostiki grippozno-bakterial'nykh pnevmonii.

A comparative examination of 226 paired sera from patients with pneumonia was carried out by CFT, HI, neuraminidase activity inhibition (NI) and double immunodiffusion. A correlation of the results of agar gel precipitation and the CFT and HI tests was observed. Convalescent sera contained antibody to influenza virus ribonucleoprotein frequently, less so to its neuraminidase or hemagglutinin. The precipitation test was shown to be highly sensitive, easy to perform, and therefore should be used in examinations of sera from patients with influenza-bacterial pneumonia.

[Analysis of influenza virus reproduction under conditions of chronic infection]. / Analiz reproduktsii virusa grippa v usloviiakh khronicheskoi infektsii

The results of comparative studies of influenza virus, original and produced in KM cells under conditions of chronic infection (KM WSN) are presented. The WSN KM virus was shown to band in the density range of 1.18 to 1.23 g/ml, with maximum at a.215 g/ml. The electrophoretic analysis of the standard virus RNA revealed mostly heavy fragments. The analysis of a total RNA preparation from the virus population produced by chronically infected cells revealed both heavy and light RNA fragments. The total synthesis of cellular DNA and RNA was slightly inhibited by KM WSN. A mechanism of persistence is suggested associated with continuous production of defective non-infectious particles.

[Comparative antigenic analysis of J-96 cell oncornavirus and Mason-Pfizer virus from a spontaneous monkey mammary tumor]. / Sravnitel'nyi antigennyi analiz onkornavirusov kletok J-96 i virusa Mézon-Pfaizera iz spontannoi opukholi molochnoi zhelezy obez'ian

A comparative study of the antigenic structure of oncornavirus of J-96 cells and Mason-Pfizer virus (M-PVM) isolated from cells of M. rhesus spontaneous mammary tumour was carried out using immunodiffusion in agar, immune autoradiography and indirect immunofluorescence procedures. Immune rabbit serum to disrupted J-96 virus detected by agar immunodiffusion in preparations or purified virus three soluble antigens one of which was identical to group-specific M-PVM antigen.

[Further study of J-96 transplantable cell oncornavirus]. / Dal'neishee izuchenie onkornavirusa perevivaemykh kletok J-96

The symplast-forming capacity of oncornavirus of continuous J-96 cells was studied by KS-test. Syncytia were formed upon combined cultivation of KS cells with J-96 cells and cells of human embryo skin-muscle tissue infected with J-96 virus as well as upon direct inoculation of KS cells with oncornavirus J-96. Formation of syncytia of KS cells correlated with the amount of J-96 cell oncornavirus. The possibility of using KS-test for titration of J-96 virus and antibody to it has been demonstrated.