[Antiviral and immunomodulating activity of polyprenyl phosphates in viral infections].

Expressed antiviral activity of Fortepren (FP) and Gamapren (GP), polyprenyl phosphate (PPP)-contaning agents, was demonstrated in experiments on mice infected with the human herpes simplex virus, type 1 (HSV1) or the vernal encephalitis virus (VEV). Since both the viral infections are of great social significance, the PPP-containing agents should be considered prospective for the medical practice. The experimental data suggested that both the drugs considerably inhibited the VEV infectiousness in the susceptible cell culture. The quantity of protein E, the main immunogen of VEV, in the culture fluid of the VEV infected cells was shown to be markedly lowered under the effect of FP and GP. It was demonstrated for the first time that FP and GP significantly inhibited evolution of the VEV protein E in the cell culture J-96. The experiments with the infectious rhinotracheitis virus (IRTV) of the corned cattle revealed that FP and GP greatly retarded the HSV1 development in the susceptible cell culture. One of the mechanisms of the antiviral action of the PPP-containing agents was likely the effect on the evolution of the virus proteins in the cells. The impact of FP on production of some key cytokines (CT) was studied on mice with experimental vernal encephalitis (IFN-gamma, IL-4 and IL-12). The content of the above mentioned CT in blood of the mice was determined by the IFA test. Under the normal conditions and in the mice infected with VEV, production of IL-12 and IFN-gamma was shown to be stimulated during the first 3-5 days after the FP administration, whereas in the animals not exposed to FP there was observed stimulation of the IL-4 production during the first 3 days after the contamination, followed by increased production of IL-12 and IFN-gamma.

[Capacity of fusicoccin for inducing the production of early interferon].

The response of human and animal cells to the action of fusicoccyne (FC), a fungal metabolite with phytohormonal properties, was evaluated. As shown by in vitro studies, FC had the capacity to induce the production of early interferon (IFN) in the blood serum of non-inbred white mice and to enhance the natural cytotoxic activity of human lymphocytes. In vitro experiments also revealed that the action of FC inhibited the metabolism of actively proliferating monocytic leukemia cells J-96 and human ovarian carcinoma cells CaOv, as well as mouse fibroblasts L-929. The common character of the mechanism of action of FC and IFN, having well-known antiproliferative and immunomodulating activity, is discussed.

[The capacity of fusicoccin for inducing the synthesis of early interferon].

The response of human and animal cells to the action of fusicoccin (FC), a fungal metabolite with phytohormonal properties, was evaluated. The capacity of FC for inducing the synthesis of early interferon (IFN), supplied into the blood serum of common white mice, and for enhancing the natural cytotoxic activity of human lymphocytes in vitro was established. The metabolism of actively proliferating monocytic leukemia cells J-96 and human ovarian carcinoma cells CaOv, as well as mouse fibroblasts L-929, was found to be inhibited under the in vitro action of FC. The common character of the mechanisms of action of FC and IFN having antiproliferative and immunomodulating activity is discussed.

Morphological and histochemical properties of human embryonic cells transformed by Rous and polyoma viruses.

It is shown that human embryonic cells transformed by Rous sarcoma virus (stable cell line 23) and those transformed by polyoma virus (stable cell line P-2) are morphologically distinguished from the normal human embryonic cells. The mitotic activity of P-2 cells was 51% and the mitotic activity of 23 cells was 48%. While the mitosis activity of human embryo fibroblast was 28%. The duration of the mitosis of P-2 cells was 20 hours and that of 23 cells was 18 hr. The duration of the mitotic cycle of human embryo fibroblast was 18 hr. The G1 periods lasted 6 hours for both the cell lines; the S period of P-2 cells lasted 8 hr and the S period of 23 cells was 6 hr. Both the cell lines had a high content of RNA, DNA, protein bound SH-groups, and a high activity of acid phosphatase, acid RNAase and glucose-6-phosphatase. The content of glycogen, and acidic mucopolysaccharides, the activity of NADPH-tetrazolium reductase, succinic dehydrogenase of both the lines were the same as in normal human cells.

Comparative cytogenetic study of a continuous human cell line and its subline susceptible and resistant, respectively, to coxsackie B3 virus.

Cytogenetic characteristics of the J-96 human cell line and its J-41 subline, highly susceptible and resistant, respectively, to coxsackie B3 virus, were compared. The J-41 subline, as compared to the original J-96 line, had fewer chromosomes in the modal class cells (54-57 and 58-62, respectively) mostly at the expense of normal chromosomes. In most J-41 cells chromosome 21 was eliminated and the number of homologues of chromosomes 2, 9, 11, and 12 was reduced. The percentage of marker chromosomes in the J-41 subline (31.3) was slightly higher than in the J-96 line (24.3). The relationship between differences in the karyotypes and properties of the cultures such as resistance to coxsackie B3 virus, capacity to produce virus-induced interferon and to acquire an antiviral state after treatment with interferon were discussed.

[Identification of genes induced to chondrocytes by nitric oxide]. / Poisk i identifikatsiia genov, indutsiruemykh v khondrotsitakh pod deistviem okisi azota.

Nitric oxide (NO) acts as a short-lived paracrine factor and selectively activates transcription of certain genes. The spectrum of inducible genes was studied in primary chondrocytes. A cDNA library was obtained by subtraction hybridization with RNAs isolated from rabbit chondrocytes before and after treatment with nitrosoglutathione, an NO-generating agent. Some of the cloned cDNAs were homologous to known mammalian genes and human EST. NO-dependent transcriptional activation was demonstrated for the stromelysin 1 and cyclooxygenase 2 genes and, for the first time, for mcl1 coding for an apoptosis suppressor.

[Induction of interferon-specific enzymes in cultures of cells sensitive and resistant to interferon]. / Induktsiia interferonspetsificheskikh fermentov v kul'turakh, chuvstvitel'nykh i rezistentnykh k interferonu.

Induction of 2'-5'-oligoadenylatesynthetase (2-5A synthetase) by interferons and theophylline by means of activation of cAMP-system in interferon susceptible and resistant cell lines were studied. In interferon resistant cell lines the basal activity of 2-5A synthetase exceeded the level of the same enzyme in interferon susceptible cell lines. Activity of 2-5A synthetase is increased in interferon susceptible cell lines by interferon treatment, but the activity of the enzyme is not altered in interferon resistant cell lines. Among the studied cell lines the induction of 2-5A synthetase by theophylline was possible only in L929 cell line. The common mechanism for the absence of 2-5A synthetase induction by interferon and theophylline in interferon resistant cells is discussed.

[Interferon-specific cellular receptors in cultured human cells, differing in sensitivity to alpha-interferon]. / Interferon-spetsificheskie kletochnye retseptory v kul'turakh chelovecheskikh kletok, razlichaiushchikhsiia po chuvstvitel'nosti k alpha-interferonu.

The interferon-specific cellular receptors in human cells cultures differing in sensitivity to alpha-interferon have been studied. The J-41 cells resistant to alpha-interferon are practically devoid of receptors highly specific to alpha-interferon. The coefficient of equilibrium and the number of receptors analyzed after Scatchard for J-96 culture of cells are 15.6 x 10(11) M-1 and 210 +/- 90, respectively. Evidently, resistance of J-41 cells to alpha-interferon is connected with the absence of interferon receptors and, as a consequence, inability to interferon-receptor interaction.

[Simultaneous identification of nucleolus organizer regions and differential chromosome staining in continuous human cell cultures]. / Odnovremennoe vyiavlenie raionov iadryshkovogo organizatora i differentsial'naia okraska khromosom v perevivaemykh kul'turakh chelovecheskikh kletok.

The application of NORs and R-bands for simultaneous staining of nucleolar organizer regions and banding chromosomes of stable human line is proposed. This technique was applied to studies on the human stable cell line J-96.

[Morphological and cytochemical characteristics of human cells transformed and made malignant by Rous and polyoma viruses]. / Morfologicheskaia i tsitokhimicheskaia kharakteristika kletok cheloveka, transformirovannykh i malignizirovannykh virusami Rausa i poliomy

In cells of human embryo skin--muscle tissue transformed by the Rouse sarcoma virus (23rd cell line) and polyoma virus (P-2 cell line), the mitotic activity was 48 0/00 for 23rd line, 51 0/00 for P-2 line as against 28 0/00 in the control cells. The transformed cells possessed greater amounts of RNA and DNA and protein--bound SH-groups, different forms of glycogen deposits, as well as higher acid phosphatase enzyme activities; there was practically no difference in acid mucopolysaccharide content or NAD-H2-diaphorase and succinate dehydrogenase activities.

[Formation of heterosymplasts in human reticular cell cultures]. / Obrazovanie geterosimplastov v kul'turakh retikuliarnykh kletok cheloveka.

The cell fusion has been studied in human reticular cell cultures J-96 and J-41 treated with the Sendai virus or with polyethylene glycol 1000 and 6000. The J-96 cells have a high alkaline phosphatase activity, in J-41 cells the enzyme is not detectable. No heterogenous alkaline phosphatase activity was seen in the protoplasm of symplasts 18 hours after virus cell fusion. It has been shown with polyethylene glycol treatment that during the fusion of cells J-96 and J-41 the enzyme activity was spreading over the symplast protoplasm.

[Heterochromatinic segments in chromosomes of cultured human cells, sensitive and resistant to Coxsackie B viruses]. / Geterokhromaticheskie raiony v khromosomakh kul'tur chelovecheskikh kletok, chuvstvitel'nykh i rezistentnykh k virusam Koksaki B.

Comparative karyological studies of C-heterochromatin have been made on line J-96 of human cells, which are susceptible to enteroviruses, and on cell line J-41 derived from this culture and possessing highly specific resistance to Coxsackie B viruses. It was shown that the development of specific resistance to Coxsackie B viruses was accompanied by the loss of one of the chromosomes of pairs 1 and 9, and by the dissapearance of two marker chromosomes. There appeared new marker chromosomes with additional C-heterochromatain regions. The data obtained are discussed with respect to a possible interrelationship between these chromosomal alterations and the specific resistance to Coxsakie B viruses.

[Relationship between interferon priming effect and functional activity of human blood cells]. / Zavisimost' praimiruiushchego éffekta interferona ot funktsional'noi aktivnosti kletok krovi cheloveka.

Relationship between the priming effects of interferon (IFN)-alpha and -gamma and the standard IFN production by peripheral blood leukocytes was evaluated in 25 volunteers (13 men and 12 women, mean age 35.3 years). Priming of IFN-alpha and IFN-gamma production positively correlated with the standard IFN-alpha and IFN-gamma synthesis, respectively. On the other hand, primed production of IFN-alpha negatively correlated with that of IFN-gamma. Possible role of types I and II IFN interactions in the maintenance of physiological regulatory balance is discussed.

[Production of J-96 cell culture chronically infected with herpes simplex virus and study of some of its properties]. / Poluchenie khronicheski infitsirovannoi virusom obychnogo gerpesa kul'tury kletok J-96 i izuchenie ee nekotorykh svoistv

Comparative effectiveness of various methods for production of chronic infection with herpes simplex virus (HSV) in J-96 cell culture: inclusion into the culture fluid of the infected cultures of immune serum to herpes virus, immune serum to HSV-infected J-96 cells and frequent changes of the medium, is described. The advantage of the second method has been found. The chronically infected culture was resistant to superinfection with the homologous virus. In passages, when the chronically infected culture produced no active infectious virus, the portion of cells containing the virus antigen was 0.8%. In this culture interferon was found in a low titer. Morphological and histochemical studies also indicated some changes in chronically infected cultures as compared with the controls, namely, an increase in the number of spindle-shaped elements, thickening of the nuclear membrane, an increase in the DNA concentration. The culture has been designated JH and is in the 220th passage now.

[Antiproliferative action of swine leukocyte interferon and of an inhibitor of interferon action]. / Antiproliferativnoe deistvie svinogo leikotsitarnogo interferona i ingibitora deistviia interferona.

The results of the study of the antiproliferative effect of swine leukocyte interferon and of an inhibitor of interferon effect in experimental mice with transplanted Krebs-2 ascitic carcinoma cells are presented. The interferon inhibitor exerted antiproliferative effect similar to that of native swine interferon. Combined use of swine interferon and interferon inhibitor did not lead to summation of the antiproliferative effect of these preparations.

[Production of inhibitors of interferon action by cell lines sensitive and insensitive to the antiviral action of interferon]. / Produktsiia ingibitorov deistviia interferona kletochnymi liniiami, chuvstvitel'nymi i nechuvstvitel'nymi k protivovirusnomu deistviiu interferona.

The capacity of a number of human and murine cell cultures to produce, upon virus induction of interferon synthesis, low molecular admixtures inhibiting interferon antiviral activity was studied. The cells resistant to interferon were found to be 8-16 times as active producers of these admixtures as sensitive cells. A possible role of the production of these admixtures in functional, i. e. not depending upon the interferon receptor gene, fluctuations of interferon inducibility and sensitivity to it is discussed.

[Relationship of the characteristics of the chromosome set in human cell cultures to the production and antiviral action of interferon]. / Sviaz' osobennostei khromosomnogo nabora kul'tur kletok cheloveka s produktsiei i antivirusnym deistviem interferona.

The role of chromosomes 2, 5, 16, and 21 in production and effect of human interferon was checked in human diploid cells, human heteroploid cells J-96 and clone J-41 thereof. The J-41 cells were found to have a lower number of chromosomes 2 as compared to the other cells under study; J-41 cells produce less interferon than the other cells. Most J-41 cells lack chromosome 21. Unlike the other two cell cultures, these cells do not produce antivirus state after treatment with interferon. The number of chromosomes 16 is larger in J-96 cells than in diploid ones, and they are less sensitive to interferon than diploid cells. The experimental results confirm the importance of chromosome 2 for coding for interferon production, other chromosomes taking part in the regulation of this process. The gene of sensitivity to interferon is localized in chromosome 21, the regulator gene coding the production of repressor of sensitivity to interferon is in chromosome 16.

[Karyotype change in human cell cultures in the process of the reversion of Coxsackie B virus sensitivity]. / Izmenenie kariotipa kul'tur chelovecheskikh kletok v protsesse reversii chuvstvitel'nosti k virusam Koksaki B.

The study was done on a subline of cells reverting to sensitivity to Coxsackie B3 virus after treatment of J-41 cells resistant to this virus with a homogenate prepared from the sensitive J-96 cell culture. Cytogenetic examinations of this cell subline showed its karyological characteristics to approach those of the sensitive J-96 culture. The modal number of chromosomes and the number of chromosomes 2, 9, 11, 12, and 21 were completely restored and marker chromosomes typical of the sensitive culture appeared. In the reverted subline there was almost no marker chromosomes peculiar for the resistant J-41 culture. In addition, a decrease in the number of chromosomes 1 and 19 replicas as compared with J-96 and J-41 culture cells and the presence of marker chromosomes not found in the original cultures indicate that this subline has its own distinctive characteristics.

[Optimal conditions for demonstrating the antimitotic action of interferon]. / Optimal'nye usloviia proiavleniia antimitoticheskogo deistviia interferona.

The effect of human leukocyte interferon (HLI) on three human cell cultures of different origin was studied. HLI inhibited the mitotic activity (MA) of human ovary carcinoma cell culture and to a lower extent that of a diploid cell culture. At the same time HLI preparation exerted a nonspecific stimulating effect on MA of the primary human embryo cell culture. The optimal conditions for the determination of the antimitotic effect of interferon on cells of tumor origin include: a seed dose of 5 X 10(4) cells/ml, addition of interferon to the culture 24 hours after cell seeding; the maximum effect of interferon is observed at 24 hours after addition to the culture.

[Cellular sensitivity to the effect of interferon during different forms of viral pathology]. / Kletochnaia chuvstvitel'nost' k deistviiu interferona pri razlichnykh formakh virusnoi patologii.

A new approach to evaluation of interferon (IFN) status of patients with acute respiratory viral infections, herpes, and urogenital infections has been developed. It consists in measurement of IFN-alpha and IFN-gamma production by leukocytes primed with IFN-alpha and IFN-gamma, respectively, and subsequent identification of groups of patients whose lymphocytes differ by their in vitro response to IFN priming: sensitive to IFN-alpha and IFN-gamma; sensitive to IFN-alpha but not to IFN-gamma; sensitive to IFN-gamma but not to IFN-alpha; not sensitive to IFN-alpha and IFN-gamma. Such analysis helps understand the mechanisms of IFN system deficiency in infectious diseases and promotes more effective IFN therapy due to selection of patients whose cells retain in vitro sensitivity to a certain IFN type.