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Biomedical subjects

A Sabó

Publications and source records attributed to A Sabó.

At least 19 recordsLinked to original sources

Antibody responses to the herpes simplex virus type 2 glycoprotein G in sera of human immunodeficiency virus-infected patients in Slovakia.

Thirty sera of human immunodeficiency virus-positive (HIV+) and 37 sera of HIV-negative (HIV-) individuals in Slovakia were tested for the presence of antibodies to herpes simplex virus type 2 (HSV-2) glycoprotein G (gG). A notable difference between the prevalence of HSV-2-specific antibodies in HIV+ and that in HIV- individuals was found (37% vs. 11%) confirming and extending previous reports that HSV-2 infection is an important risk factor for HIV transmission. Efforts toward the detection of HSV-2 infection and its therapy by anti-HSV drugs should be considered an important factor in decreasing the risk of contracting and spreading of HIV in Slovakia.

Animals↗

Herpes simplex virus type 1 envelope subunit vaccine not only protects against lethal virus challenge, but also may restrict latency and virus reactivation.

A subunit vaccine containing the main antigenic components of herpes simplex virus 1 (HSV-1) was tested in Balb/c mice and albino rabbits. The mice were completely protected against challenge with 10 LD50 of the highly pathogenic SC16 strain given by intraperitoneal (ip) route when immunized with 1000 antigen units (ELISA) corresponding to 110 micrograms of protein. The animals were protected against lethal disease when immunized with 1-33 micrograms of protein per dose. Immunization of rabbits with 3000 antigen units prior to inoculation of strain Kupka into right scarified cornea limited the establishment of latency in the trigeminal ganglia. Both the number of animals in which latency had been established as well as the number of homolateral sensory ganglion cells which had become virus carriers were reduced. However, the effect of immunization was less striking at preventing HSV reactivation in rabbits vaccinated after infection. When shedding of reactivated HSV was elicited by repeated epinephrine iontophoresis to cornea, there was no quantitative difference between the immunized and mock-immunized groups, only the period between stimulation and the onset of virus shedding was prolonged in immunized rabbits (from 3.6 to 5.6 days, p < 0.05). But if the corneas were stimulated by a single iontophoresis procedure, the duration of virus shedding was significantly reduced from 5.6 days in the mock-immunized rabbits to 1.7 days in the immunized ones (p < 0.025). In the latter experiment, the total number of positive swabs during 14 days of the post-stimulation period was higher in the mock-immunized animals (31 of 171, 18.1%) than in the immunized ones (12 of 162, 7.4%; p < 0.025).

Animals↗

[The immunogenicity of a vaccine against enzootic abortion in sheep].

Immunogenicity of vaccine against enzootic abortion of ewes (EPO) was evaluated in sheep and laboratory white mice. The vaccine contained purified and formalin-inactivated corpuscules of Chlamydia psittaci. Experiment was performed on ten sheep, Slovak Merino breed, which were negative before vaccination in serological assays and blastic transformation of lymphocytes (BTL) tests. The animals were immunized subcutaneously with 2 ml of vaccine which contained 100 micrograms (sheep No. 1-4) and 20 micrograms (sheep No. 5-8) corpuscules of C. psittaci. Control group (No. 9-10) received the same volume of physiological saline. The second dose of vaccine was given one month after the first dose (No. 1-2--100 micrograms each, and No. 5-6--20 micrograms each). Blood for serological evaluation and BTL test was taken before vaccination and 1, 3, and 6 months after vaccination. The sheep which were given the second dose of vaccine were also evaluated two weeks thereafter. Antibody response in complement fixation reaction (CFR) and enzyme-linked immunosorbent assay (ELISA) was compared with the results of BTL. While the antibody response evaluated by CFR was sporadically positive after administration of the higher dose of vaccine only, in ELISA all sera were positive except one lower dose of vaccine (Tab. I). In all post-vaccination intervals positivity was confirmed by BTL test irrespective of the size and number of vaccine dose (Tab. II). Immunization of mice with one dose (100 micrograms) of vaccine significantly but not completely reduced multiplication of C. psittaci in the lungs of mice (Tab. III).

Abortion, Septic↗

Non-viraemic transmission of tick-borne encephalitis virus: a mechanism for arbovirus survival in nature.

The vectors of arthropod-borne viruses (arboviruses) become infected by feeding on the viraemic blood of an infected animal. This theory is based on transmission studies involving artificial infection of vertebrate hosts by syringe inoculation. To reproduce natural conditions of virus transmission, infected and uninfected vectors (ticks) of tick-borne encephalitis virus, the most important arbovirus in Europe, were allowed to feed together on uninfected wild vertebrate hosts. The greatest numbers of infected ticks were obtained from susceptible host species that had undetectable or very low levels of viraemia. The results suggest that 'nonviremic transmission' is an important mechanism for the survival of certain arboviruses in nature.

Animals↗

Herpes simplex virus type 2 and pseudorabies virus associated growth factors and their role in the latency in vitro.

A putative herpes simplex virus type 2 (HSV-2) growth factor (HSGF-2) was detected in a crude extract from virus infected mouse embryo cells. This factor, similar to previously described pseudorabies virus (PRV) associated growth factor (PRGF) was shown to have ability to morphologically transform non-transformed cells and to repress the transformed phenotype of transformed cells. Both activities could be neutralized with two, out of seven monoclonal antibodies directed against glycoprotein B of HSV-2. Both PRGF and HSGF-2 were detected in human embryo lung cells latently infected with PRV or HSV-2 either at 41 degrees C, or in the presence of phosphonoacetic acid. Human alpha-2 interferon, when present in medium of latently infected cells enhanced the production of both HSGF and PRGF. On the contrary, when latently infected cells were treated with 5-azacytidine the synthesis of both PRGF and HSGF-2 was completely blocked and the virus reactivated from latency replicated to higher titers than in non-treated cells. The role of PRGF and HSGF-2 in the establishment, maintenance and reactivation of latency, as well as in cellular transformation is discussed.

Animals↗

Transforming activity of crude extract of pseudorabies virus-transformed cells.

Crude extract of pseudorabies virus (PRV)-transformed human (H-PR-1) cells induced transformation in human embryonic lung (HEL) cells. When the extract was removed, the acquired cell morphology remained unchanged, but the saturation density of cells was decreased. The transforming effect of the extract was neutralized with anti-PRV IgG.

Cell Transformation, Viral↗

Analysis of reactivation of latent pseudorabies virus infection in tonsils and Gasserian ganglia of pigs.

Multiplication of pseudorabies virus (PRV) strain TOP and establishment of latency were followed in the tonsils and Gasserian ganglia (GG) of 8-week-old pigs after oral infection. Already within 24 hr, the titre of PRV in tonsils reached 10(2) TCID50/g tissue, its presence in the oropharynx being detectable by swabbing. Virus multiplication in tonsils culminated on days 3-6 p.i. (10(6)-10(6.5) TCID50/g) and continued till day 11 post-infection (p.i.), when its titre was 10(1.5) TCID50/g the swabs being still positive. The presence of the virus in GG was first proved at 48 hr p.i., but no virus could be found there by titration on day 11. The virus titre in the GG had essentially followed that in tonsils, however, it was substantially lower during the whole experiment. After primoinfection no shedding of PRV was detected by oropharyngeal swabs taken at 3-5 day intervals during the period of 360 days. Nevertheless, explantation of tonsils and GG, performed between days 60-360 p.i., revealed the presence of PRV latency in 41.1 per cent of animals. The total activation rate in GG cultures was 71.5 per cent, while in the cultures of tonsils 28.5 per cent only. Cocultivation of cell suspensions obtained by trypsinization of GG and tonsillar tissue with chick embryo cells (CEC) did not result in detection of the latent virus though latency was confirmed by explantation of the same tissue samples. However, already 24 hr in culture were sufficient for activation of latency as judged by trypsinization of the explanted tonsillar and GG fragments and their inoculation onto CEC. Estimation of the number of cells carrying PRV in the GG showed that activation occurred in one out of 10(4)-10(5) ganglion cells.

Animals↗

Vesicular stomatitis virus phenotypically mixed with retroviruses: an efficient detection method.

Two methods of assaying vesicular stomatitis virus (VSV) particles phenotypically mixed with retrovirus-coded antigens were compared. Each of them detected phenotypically mixed particles with different minimum proportion of surface glycoprotein molecules of the donor virus, and consequently also profoundly different proportions of VSV virions containing retrovirus antigens. Only a low proportion (10(-4) of VSV virions grown in XMuLV-infected rabbit SIRC cells behaved as pseudotypes, resistant to anti-VSV serum and neutralized by anti-XMuLV serum. VSV produced in mouse L cells did not contain significant titre of pseudotype particles in the neutralization test. However, when immunoprecipitation was used with corresponding antibody and Staphylococcus aureus cells, almost 100% of the VSV virions produced in L cells and in XMuLV-preinfected SIRC cells were found to contain MuLV-related antigen molecules.

Animals↗

The effect of antibody on latent pseudorabies virus infection in vitro.

Cytosine arabinoside (Ara C) inhibited the synthesis of pseudorabies virus when Vero cells were infected at a multiplicity of infection of 0.0001-0.05 PFU per cell. On removal of Ara C, infectious virus reappeared after a latent period of 3-5 days. The activation of latent virus was not influenced by elevating the temperature to 40 degrees C at the time of Ara C removal but it was prevented by antiviral antibody. When antiviral IgG was added into the culture fluid of cells either during the incubation with Ara C, or after removal of the inhibitor, the number of infectious centres was reduced to about 10%. The role of antiviral IgG in the maintenance of latency is discussed.

Animals↗

Latent pseudorabies virus infection established at supraoptimal temperature.

Little or no infectious virus was recovered from BHK-21 cells adapted to 40 degrees C, when they were infected with an attenuated strain of pseudorabies virus (PRV) at low multiplicity of infection (MOI) and subsequently kept at 40 degrees C. By passaging the infected cells at 40 degrees C, infectious virus disappeared within 2-3 passages. When the cells were shifted down to 37 degrees C, activation of virus growth occurred after a latent period of 48-72 hr. Infected cells kept at 40 degrees C were as sensitive to superinfection with the virulent PRV strain TOP as the control cells. The ability of PRV strains to enter into the described latent state was related with the degree of their virulence. When cells were treated with 5-bromo-2-deoxyuridine (BUdR) at 40 degrees C for 24 hr and then shifted to 37 degrees C, a slight increase in the number of infectious centres was observed and the latent period was also prolonged.

Animals↗

Activation of latent herpesvirus hominis in explants of rabbit trigeminal ganglia: the influence of immune serum.

More than fifty albino rabbits were inoculated into the right scarified cornea with 10(7) PFU of the Kupka strain of human herpes virus type 1 (HHV-1). At intervals ranging from 4--280 days post infection (p.i.), both gasserian ganglia, both trigeminal nerve trunks and pieces from brain stem and from both corneas were explanted. Activation of the latent HHV-1 was found mainly in the homolateral ganglion tissue, but also in explants originating from the opposite ganglia. Within 24--72 hours, prior to the release of virus into the medium, one infectious unit of HHV was recovered from 10(4)--10(5) cells of the ganglion explant. In addition, a few neurons and satellite cells revealed the presence of virus-specific antigens when the explants were examined by immunofluorescence in serial sections. If the gangia were explanted in the presence of immune serum, the virus recovery rate was at least twice lower as compared to the virus activation in explants kept in the absence of immune serum.

Animals↗

Susceptibility of various cell lines to virulent an attenuated strains of pseudorabies virus at supraoptimal temperature.

Replication of virulent and attenuated strains of pseudorabies virus (PRV) at supraoptimal temperature was studied in rabbit lung (ZP), pig kidney (PS), and BHK-21 cells adapted and non-adapted to 40 degrees C and L, HeLa and human amniotic (Am) cells non-adapted to 40 degrees C. The temperature of 40 degreeC did not influence the type cytopathic effect (CPE) in either adapted or non-adapted cells. According to the susceptibility at 40 degrees C the cells could be divided into three groups: cells permissive for all PRV strains (adapted and non-adapted ZP and non-adapted BHK-21 and PS); cells non-permissive for all PRV strains (L, HeLa and Am); and cells permissive for virulent and non-permissive for attenuated PRV strains (adapted BHK-21 and PS). The virus titres in permissive cells at 40 degrees C did not differ from those obtained at 37 degrees C. The attenuated strain CK-PRV X was found to be a tsmutant of PRV with host range character.

Cell Line↗

The effect of supraoptimal temperature on the formation of pseudorabies virus particles.

Replication of virulent and attenuated strains of pseudorabies virus (PRV) at supraoptimal temperature was inhibited in L cells non-adapted to 40 degrees C. Electron microscopy revealed the prevalence of hollow core particles without nucleoid; both the nuclei and cytoplasm of L cells were markedly altered at supraoptimal temperature. In BHK-21 cells adapted to 40 degrees C only replication of attenuated strains was inhibited whereas the virulent ones reached high titres at this temperature. The particles produced at 40 degrees C by attenuated strains were non-infectious but morphologically normal. The ability of attenuated PRV strains to reproduce in adapted BHK-21 cells at 40 degrees C was in correlation with the degree of their attenuation. Changes in the nuclei and cytoplasm of adapted BHK-21 cells at supraoptimal temperature were not as marked as in L cells.

Animals↗

Susceptibility of various cell lines to virulent and attenuated strains of pseudorabies virus.

Various cell lines were infected with virulent and attenuated strains of pseudorabies virus (PRV). According to the type of the cytopathic effect (CPE), the cells could be divided into 3 groups: cells in which all strains formed syncytia; cells in which all strains caused rounding of cells; and cells in which virulent strains caused syncytium formation while attenuated ones rounding of cells. Neither the form (fibroblastoid or epithelial) nor the origin of cells influenced the type of the CPE. L cells and some cell lines derived from human tissue proved to be less sensitive to PRV than other cells. In certain human cells (e.g. HeLa and D6) virulent PRV strains propagated better than attenuated ones.

Cell Line↗

Latent pseudorabies virus infection in pigs.

Latent infection with the TOP strain of pseudorabies virus (PRV) was established in 6 weeks old piglets. Infectious virus was found in oropharyngeal swabs till day 10 post infection (p.i.); later on, attempts to detect the virus remained unsuccessful. However, PRV could be isolated by explantation of tonsils, cervical lymph nodes, nasal mucosa and gasserian ganglia. These tissues were removed between 160 and 181 days p.i., during cultivation, infectious PRV was released into the culture fluid from the 3rd to the 11th day of the explantation. PRV antigen was seen by immunofluorescence only in explants coming from gasserian ganglia. It was localized in both neurons and satellite cells. In piglets given hydrocortisone before explantation, the number of virus-producing explants was not enhanced as compared to that of virus-producing explants from untreated animals.

Animals↗

Experimental latent herpesvirus infection in rabbits, mice and hamsters: ultrastructure of the virus activation in explanted gasseric ganglia.

The frequency of latent infection as established in trigeminal ganglia of rabbits, mice and hamsters with human herpesvirus type 1 (HVH) was compared using two different virus strains. Explantation proved to be effective in reisolation of HVH from ganglion tissue, which did not yield infectious virus at time of its removal. After healing of acute keratitis, the latent infection in homolateral gasseric ganglia of rabbits was detected at a relatively high frequency (60-80 per cent) up to 120 days post infection (p.i.) in case of both virus strains. The activation rate was a little lower in hamsters. After inoculation of suckling and young mice with a sublethal dose of HVH by oral and nasal routes, approximately 40-100 per cent of the animals had virus in their gasseric ganglia during the acute period; 30-60 days later only 10-25 per cent had virus in the latent form. Immunofluorescent and electron microscopic examination of the explanted ganglion tissue showed the presence of HVH in neurons, neuronal satellites and Schwann cells. The nuclei of noneural cells contained numerous crystalline arrays. The possibility that pseudounipolar neurons of the regional sensoric ganglion are not the exclusive site of HVH latency is discussed.

Animals↗