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Biomedical subjects

V Vonka

Publications and source records attributed to V Vonka.

227 records · Page 13Linked to original sources

Thermoinactivation of human cytomegalovirus.

Vonka, Vladimir (Baylor University College of Medicine, Houston, Tex.), and Matilda Benyesh-Melnick. Thermoinactivation of human cytomegalovirus. J. Bacteriol. 91:221-226. 1966.-The inactivation at 4 and 37 C of several strains of human cytomegalovirus was studied. The preliminary findings that freshly harvested cytomegalovirus was inactivated more rapidly at 4 C than at higher temperatures was confirmed. Intracellular virus still within infected cells was found to be more stable at 4 C than virus released by sonic treatment just before incubation at 4 C. The composition of the diluent played an important role. In tris(hydroxymethyl)-aminomethane buffer, virus was unstable at both 4 and 37 C, with the rate of inactivation faster at 4 than at 37 C. Similar results were obtained when bicarbonate-phosphate buffer or Eagle's medium when bicarbonate was used as virus diluent. Calf serum stabilized the virus at 37 C, but not at 4 C. The deletion of bicarbonate from Eagle's medium had a stabilizing effect at both temperatures. An even greater stabilizing effect at both 4 and 37 C was obtained when distilled water was used as virus diluent. Inactivation rates varied from one strain to the next at 4 C but not at 37 C. Differences were found also with virus progeny derived from a single strain, but harvested at different stages during virus multiplication. Virus harvested early was more labile at 4 than at 37 C, whereas the late virus was more labile at the higher temperature. Intracellular and extracellular virus preparations were inactivated at the same rates at either 4 or 37 C.

Culture Media↗

Correlation between infectivity and physical virus particles in human cytomegalovirus.

Benyesh-Melnick, Matilda (Baylor University College of Medicine, Houston, Tex.), Fern Probstmeyer, Robert McCombs, Jean P. Brunschwig, and Vladimir Vonka. Correlation between infectivity and physical virus particles in human cyto-megalovirus. J. Bacteriol. 92:1555-1561. 1966.-Infectivity titers [measured as plaque-forming units (PFU)] and particle counts by the sedimentation pseudo-replication technique were determined for crude, unpurified, intracellular preparations of two different strains of human cytomegalovirus. Unlike the high particle-infectivity ratio of 10(6) to 10(8) previously reported for these viruses, the number of total particles per PFU ranged from 160 to 490 with strain AD-169 and from 176 to 1,050 for strain C-87. Interpretation of particle-PFU ratios of intracellular cytomegalovirus in terms of particle morphology is not conclusive at this time. The number of enveloped forms found varied between 0 and 34% of the total particles counted. However, the true proportion is probably greater, because envelopes were found to be destroyed by the enzyme treatment used in preparing the specimens for examination in the electron microscope. The number of full particles found ranged between 4 and 31% of the total particles counted. The particle per PFU ratio of extracellular virus was found to be three- to fivefold lower than that of intracellular virus.

Cytomegalovirus↗

DNA vaccine against oncogenic hamster cells transformed by HPV16 E6/E7 oncogenes and the activated ras oncogene.

The capability of DNA to elicit anti-tumour immunity was studied using human papillomavirus type 16 (HPV16)-transformed Syrian hamster cells denoted K3/II. These cells had been derived after cotransfection of primary kidney cell cultures with p16HHMo plasmid containing E6/E7 oncogenes of HPV16 and pEJ6.6 plasmid containing the activated human H-ras oncogene; they express both the HPV16 and activated H-ras genes. As a DNA vaccine, the p16HHMo plasmid was used. Three doses of the plasmid (either 100 microg or 10-15 microg per dose) were administered intramuscularly at 3-week intervals. The animals were challenged with four different doses (10(3)-10(6) per animal) of K3/II cells 10 days after the last plasmid injection. In one experiment the lower dose of plasmid DNA was also given in a mixture with the cationic lipid DOTAP. In another experiment, the pEJ6.6 plasmid (100 microg per dose) was used either alone or in combination with p16HHMo. In all experiments animals inoculated with the same doses of pBR322 plasmid served as controls. A moderate protective effect was observed in animals inoculated with the 100-microg doses of p16HHMo, but not in those inoculated with 10-15 microg of the same plasmid, whether given with or without DOTAP. A protective effect was also observed after administration of the pEJ6. 6 plasmid. At the time of challenge a portion of the p16HHMo-immunized, but not the pBR322-treated, animals possessed antibodies reactive in ELISA with peptides derived from the N-terminal portion of HPV16 E7 protein and with one peptide derived from E6 protein, while two other E6 peptides exhibited non-specific reactivity.

Animals↗

Chemoimmunotherapy of cancer: potentiated effectiveness of granulocyte-macrophage colony-stimulating factor and ifosfamide derivative CBM-4A.

The effectiveness of combined chemoimmunotherapy with ifosfamide derivative CBM-4A and granulocyte-macrophage colony-stimulating factor (GM-CSF) was investigated in two experimental tumor models, 3MC-induced MHC class I+ sarcoma Mc12 and HPV16 E6/E7 oncogene-induced MHC class I- carcinoma MK16, transplanted in syngeneic mice. Treatment of Mc12 and MK16 tumor-bearing mice with GM-CSF or CBM-4A alone produced moderate anti-tumor effects. However, when the tumor-bearing mice were first treated i.p. with a single dose of CBM-4A (150 mg/kg) and three days later peritumorally with five daily doses of GM-CSF (100 ng/day), substantially stronger tumor-inhibitory effects were observed. The results indicate that in both, MHC class I+ and MHC class I- tumors, the combined chemoimmunotherapy can inhibit tumor progression more effectively than GM-CSF therapy or chemotherapy alone, and they suggest that GM-CSF should be considered as adjuvant to chemotherapy in clinical trials with HPV 16-associated neoplasms.

Animals↗

[Mechanism of penetration of human type 3 parainfluenza virus into monkey kidney cells].

The mechanism of human parainfluenza type 3 virus penetration into monkey kidney cells was studied by morphological and biochemical methods. The results of electron microscopic studies permit a conclusion that the virus penetrates into the cells by the mechanism of receptor endocytosis. Analysis of subvirus structures in cytosole revealed two types of particles: nucleocapsids and structures of a larger size and lower buoyant density containing, in addition to NP protein, matrix (M) protein. It is presumed that nucleocapsid is released from the endocytic vacuole into the cytosole in association with M protein which is gradually eliminated from the nucleocapsid surface.

Animals↗

Small-scale field trial with neuraminidase vaccine.

An inactivated whole-virus vaccine was prepared from the influenza virus recombinant H0N2 and administered to two groups of subjects. A total of 1,200 subjects were vaccinated; comparable groups of subjects served as controls. From a portion of the vaccinees sera were obtained prior to vaccination and three to four weeks afterwards. Serological tests revealed development of or increase in the antibody against H0 in a great majority of the vaccinated subjects and against N2 in slightly more than half of them. Antibody response to H3 antigen was only rarely encountered. Approximately three months after vaccination an influenza epidemic caused by Victoria-like (H3N2) viruses broke out in Czechoslovakia. Numerous influenza cases occurred in the two populations followed. Morbidity was significantly lower among the vaccinated than among the control subjects, indicating a protective effect of the neuraminidase vaccine under field conditions.

Adolescent↗

Antibody response in humans after administration of whole-virion and split vaccine prepared from two different influenza A/swine viruses.

Previous findings with sera of several different animal species had indicated a close antigenic similarity between the X53 recombinant (possessing both surface antigens of A/Swine/New Jersey/76 virus) and the A/Swine/Ann Arbor/31 (AA) virus (1). Since the latter virus grows much better in chick embryos than the former, we examined the possibility of using the Ann Arbor virus instead of X53 for inactivated vaccine production. The present paper describes our findings in sera of subjects immunized with either whole-virus or split vaccine prepared from AA virus.

Adolescent↗

Antibody formation in laboratory animals after parenteral application of A/swine influenza virus.

Antigenicity of swine influenza viruses, Ann Arbor/31 and recombinant X-53, was compared on the different animal species. The serological investigations of animals infected with live or immunized with the inactivated virus, demonstrated a close antigenic relationship between the H and N antigens of both viruses. For inactivated vaccine preparation it was recommended, on the basis of our results, to use Ann Arbor virus instead of virus X-53.

Animals↗

Properties of Syrian hamster cells transformed by human papillomavirus type 16.

Adult Syrian hamster kidney cells were transfected with a mixture of plasmids containing human papillomavirus type 16 (HPV16) E6/E7 open reading frames (ORFs), activated Ha-ras gene and neomycin resistance gene. From these cultures two lines were isolated which were oncogenic for newborn and 5-day-old but not for 3-week-old hamsters. Sublines oncogenic for 3-7-week-old hamsters were derived from tumours formed in animals inoculated within 5 days of birth. The cells contained HPV 16 DNA in an integrated form and HPV16 transcripts. The transcript patterns in low and high oncogenicity sublines were different. Very few tumour-bearing animals possessed antibodies reactive with E6- and E7-derived synthetic peptides. On the other hand a majority of these animals gave a positive reaction in the lymphoproliferation assay with either E6 and E7 peptides or extracts from the transformed cells.

Animals↗