[Production of the monoclonal antibodies to the rabies virus nucleoprotein].

Five hybridomas secreting monoclonal antibodies (MAbs) for the nucleocapsid protein of the rabies virus were obtained through the fusion of the SP2/0 murine myeloma cells with splenocytes of BALB/c mice immunized with fixed rabies virus (CVS strain). All hybridomas secret MAbs of the IgG class that display different specificity to the nucleocapsids of rabies and rabies-related viruses. MAbs 2ell showed the specificity for the prevalent in Russia rabies viruses that are similar to commercially available anti-rabies conjugate.

[Development of a diagnostic test system on the basis of sandwich ELISA for the detection of avian influenza A virus].

A panel of hybridomas producing monoclonal antibodies (MAbs) to nucleocapsid protein (NP) of avian influenza A virus was obtained. On the basis of 2 MAbs, the authors designed an antigen-bound ELISA (sandwich ELISA), in which NP3 MAbs were used as antigen-bound antibodies and NP MAbs conjugated with horse radish peroxidase as antigen detection antibodies. The specificity of the test system to avian influenza virus was determined. The developed test system was ascertained to specifically detect influenza A virus of all study subtypes and to yield no cross reactions with other tested virus pathogens. The sensitivity of the sandwich ELISA was 30 ng/ml of NP in the urine-treated virus preparations. The assay was tested on experimental H5N1-infected mice. The findings positively correlated with the results of postmortem studies and with the virus isolation method in the chick embryos. The developed test system may be used to detect avian influenza A virus as an alternative or supplement to other diagnostic techniques.

[Preparation and characteristics of monoclonal antibodies to protein E2 (GP55) of classical hog cholera virus].

Twenty-eight hybridomas producing monoclonal antibodies (MAbs) to proteins of classical swine virus (CSFV) were obtained by fusion of AS2/0 murine myeloma cells with splenocytes of BALB/c mice. The recombinant E2 glycoprotein of CSFV and the gradient-purified CSFV strain Shimen were used as an antigen for immunization. Twenty-four hybridomas produced MAbs of class IgG and four hybridomas did MAbs of class IgM. All MAbs were specific for E2 protein of CSFV. Competitive enzyme immunoassay showed that MAbs detected 8 epitopes on protein E2.

[Investigation of antiviral properties of new formulations of Panavir in the treatment of experimental herpes virus infection in mice].

Protective action of Panavir was studied in the treatment of experimental herpes virus infection on albino mice. It was shown, that intravenous and rectal Panavir formulations were able to increase survival of the experimental animals (decreased lethality) infected by HSV-1.

[Comparison of adjuvant activities of glucosaminyl-muramyl dipeptide and of the gene coding for granulocyte-macrophage colony-stimulating factor in DNA immunization against herpes simplex virus].

Adjuvant activities of granulocyte-macrophage colony-stimulating factor (GM-CSF) and synthetic glucosaminyl-muramyl dipeptide (GMDP) were studied in immunization against type 1 herpes simplex virus (HSV1). Gene encoding the gD HSV1 protein (pDNAgD) was used as an immunogen. Gene encoding GM-CSF in pDNAGM-CSF plasmid, which was developed for eukaryotic expression, and GM-DP were used as immune response modulators. GMDP and plasmid DNA with inserted GM-CSF gene enhanced T-cell immune response to HSV1 after a single injection (pDNAGM-CSF) or 24 h before (GMDP) immunization with the gD HSV1 gene. Both adjuvants increased protective effect of DNA-immunization by a virus gene with 63 up to 100% after injection of two genes and up to 96% after the viral gene was inoculated 24 h after GMDP. These high effects indicate that further investigation of anti-HSV1 DNA-based vaccines used with genetic and peptide adjuvant is prospective.

[Effect of tumor necrosis factor DNA on immune response to DNA immunization against herpes simplex virus]. / Vliianie DNK faktora nekroza opukholi na immunnyi otvet pri DNK-immunizatsii protiv virusa prostogo gerpesa.

A study was made of the adjuvant effect of the mouse tumor necrosis factor alpha (mTNF alpha) on DNA immunization against the herpes simplex virus type 1 (HSV1). The HSV1 gD gene (pDNAgD) served as an immunogen; mTNF alpha or its gene cloned in an eukaryotic expression vector (pDNAmTNF) were used to modulate the immune response. Double immunization with pDNAgD led to a sixfold increase in the in vitro T-cell response, a high (1:2000) titer of anti-HSV1 antibodies (including virus-neutralizing antibodies), an increase in IgG2a/IgG1 (suggesting a shift of the immune response to the Th1 type), and no change in CD4/CD8 T-cell ratio. A single injection of mTNF alpha along with inactivated HSV1 allowed a twice higher antibody titer and a fourfold higher T-cell response as compared with immunization with HSV1 alone. Double immunization with both pDNAgD and pDNAmTNF increased the titer of anti-HSV1 antibodies and the T-cell response by factors of 8 and 1.5, respectively, as compared with immunization with pDNAgD alone. However, the protective effect was significantly lower with the two plasmids than with pDNAgD (73 vs. 100%). Thus, DNA immunization with pDNAgD induced both B- and T-cell responses and completely protected mice from a lethal doze of HSV1. The adjuvant properties of mTNF alpha and pDNAmTNF need further investigation.

[Nociceptive reactions during stimulation of immunity in rats with various individual resistance to stress]. / Notsitseptivnye reaktsii pri stimuliatsii immuniteta u krys s razlichnoi individual'noi ustoichivost'iu k stressu.

Various behavioural nociceptive reactions and individual resistance against stress were studied under conditions of stimulation of the immune processes by various techniques. The research problems included a study of influence of the immune stimulation with preparation "Imunofan" upon pain responses depending on individual resistance of animals to a stress, and the obtained results were compared with similar data in natural model of immune activation. To reveal central immune regulation of nociceptive reactions, imunofan was injected into brain ventricles. The work was carried out in 43 "Wistar" adult male rats. Free "open field" behaviour of animals was recorded to define a stress-resistance. Following nociceptive reactions, tail-flick to thermal stimuli; start, escape, jumping and vocalization to electrical skin stimulation, were studied. It was shown that intramuscular injection of imunofan (0.01 ml, 0.005% solution) depressed an active behaviour of animals in open field and reduced pain thresholds. This hyperalgesia was much higher in non-resistant rats in comparison to the resistant ones. Similar results were obtained in natural activation of immunity caused by operative procedure necessary for injection of imunofan into ventricles. Intracerebroventricular injections were accompanied by stronger and more complex changes of pain sensitivity.

[Morphine antibodies in the mechanisms of morphine resistance, morphine tolerance and of the action of an antagonist]. / Antitela k morfinu v mekhanizmakh morfinnoi rezistentnosti, morfinnoi tolerantnosti i deistviia antagonista.

It was found that the absence of the analgesic effect of morphine, as determined by the tail-flick test, in morphine-resistant and morphine-tolerant rats, as well as in naloxane blockade of morphine analgesia in morphine-sensitive rats was attended with a four- to eight-fold increase in morphine antibodies in the plasma, as determined by the ELISA method. It is suggested that a pharmacokinetic factor mediated through immune reactions of morphine antibodies formation is one of the mechanisms of such conditions.

[Enkephalinase mechanisms of tolerance of analgetic action of opioids. Effects of D-phenylalanine and naloxone]. / Enkefalinaznye mekhanizmy tolerantnosti k anal'geticheskomu deistviiu opioidov. Effekty D-fenilalanina i naloksona.

The paper reviews the data available in the literature and the authors own data demonstrating the differences in the levels of endogenous opioids and in the effects of enkephalinase inhibitor and naloxone in morphine-responsive and morphine-resistant and -tolerant animals. In the morphine-tolerant animals, a single administration of enkephalinase inhibitor or some doses of naloxone was found to produce an analgesic effect leading to a short-term analgesic effect of morphine. Chronic administration of naloxone causing a gradual decrease in and subsequent cessation of its analgesic effect leads to recovery of morphine's analgesic effect in the drug-resistant and -tolerant animals. It is suggested that the morphine-resistant animals have a congenital high activity of enkephalinase, while the drug-tolerant ones have its acquired high activity. Naloxone in certain doses with the high activity of enkephalinase act as its inhibitor, but in high doses acts as its opioid antagonist.

[The enkephalinase mechanisms of the resistance and tolerance to the analgesic effect of morphine in rats. Differences in the effects of the action of D-phenylalanine in morphine-sensitive, morphine-tolerant and morphine-resistant rats]. / Enkefalinaznye mekhanizmy rezistentnosti i tolerantnosti k analgeticheskomu éffektu morfina u krys. Razlichiia éffektov deistviia D-fenilalanina u morfinchuvstvitel'nykh, morfintolerantnykh i morfinrezistentnykh krys.

In morphine-sensitive (s.c. 1.5 mg/kg) Wistar rats (60%) i.p. inoculation of 300-600 mg/kg d-Phenylalanine (d-Pha) did not change the nociception (tail-flick test), but in morphine-resistant rats (40%) evoked a dose-dependent analgetic effect. In morphine-sensitive rats (40%) chronic morphine administration induced the tolerance and d-Pha injection evoked analgetic effect. Morphine injection just after d-Pha analgesia was over evoked analgetic effect in morphine-resistant and -tolerant rats. It is suggested that morphine-resistant rats have a congenital and morphine-tolerant rats an acquired high level of enkephalinase activity which blocked the morphine analgetic action.

[The enkephalinase mechanisms of the resistance and tolerance to the analgesic effect of morphine in rats. Differences in the effects of naloxone in morphine-sensitive, morphine-resistant and morphine-tolerant rats]. / Enkefalinaznye mekhanizmy rezistentnosti i tolerantnosti k analgeticheskomu éffektu morfina u krys. Razlichiia éffektov naloksona u morfinchuvstvitel'nykh, morfinrezistentnykh i morfintolerantnykh krys.

In morphine-sensitive (s. c. 1.5 mg/kg) Wistar rats i.p. injection of 0.3 mg/kg naloxone either did not change nociception (tail-flick test) or induced hyperalgesia. In morphine-resistant rats 0.2-0.7 mg/kg naloxone injection induced analgetic effect but 1.0 mg/kg induced hyperalgesia. In morphine-sensitive rats chronic morphine administration induced the tolerance and naloxone injection evoked analgetic effect. Morphine inoculation just after naloxone analgesia was over induced analgetic effect. In morphine-resistant and -tolerant rats chronic naloxone administration induced gradual decrease and subsequent disappearance of its analgetic effect and subsequent morphine injections induced analgetic effect for some days. It is suggested that naloxone in morphine-resistant and -tolerant rats with high level of enkephalinase activity functions as its inhibitor. Chronic naloxone administration evoked progressive inhibition of enkephalinase activity that induced the morphine analgetic effect in morphine-resistant and -tolerant rats.

[Action of an enkephalinase blocker on the effect of acupuncture in acupuncture sensitive and resistant rabbits]. / Vliianie blokatora énkefalinazy na éffekty akupunktury u akupunkturo-chuvstvitel'nykh i rezistentnykh krolikov.

In unanaesthetized acupuncture-sensitive rabbit d-phenylalanine injection didn't change the EP in response to tooth pulp electrostimulation, but prolonged the analgetic effect of auriculo-acupuncture stimulation 15 Hz expressed by decreasing of the amplitude of N1P2 component EP. In acupuncture-resistant rabbit d-phenylalanine injection induced analgetic effect which was enhanced and prolonged by auriculo-acupuncture stimulation. It's suggested that the recovery of pain sensibility after acupuncture analgesia is determined by enkephalinase's mechanism activation which is activated permanently in acupuncture-resistant rabbits.