V Hronovský’s scientific contributions

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Publications (24)


Use of hyperimmune mouse ascitic fluids for arbovirus differentiation by indirect immunofluorescence and conventional serology
  • Article

October 1981

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17 Reads

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5 Citations

Acta Virologica

V Hronovský

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R Benda

The cross-reactivity of 22 arbovirus species (alphaviruses; tick- and mosquito-borne flaviviruses; and selected bunyaviruses) was tested with monovalent immune mouse ascitic fluids by indirect immunofluorescence (IIF) in comparison with classical serological reactions (virus neutralization -- VN; haemagglutination inhibition -- HI; and complement fixation -- CF -- reactions). Known relationships within the virus groups studied were confirmed. As to the differentiation limits, the VN test was followed by IIF. Evaluation of the ratio of heterologous to homologous antibody activities showed that with the exception of antigenically closely related arboviruses (Western equine encephalomyelitis -- Sindbis; Japanese encephalitis -- Murray Valley encephalitis; dengue viruses; California -- Tahyna), most arboviruses within the antigenic complexes tested could be relatively reliably differentiated by IIF.


CV-1 monkey kidney cell line: A highly susceptible substrate for diagnosis and study of arboviruses

April 1978

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98 Reads

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10 Citations

Acta Virologica

The CV-1 cell line, derived from Cercopithecus aethiops monkey kidneys, acquired in the course of serial passaging an epitheloid morphology and heteroploid karyotype. As compared with Vero and GMK cell lines, the CV-1 line proved to be more susceptible to a broad spectrum of arboviruses belonging to the families Togaviridae and Bunyaviridae. Most of them reached high titres and were markedly cytopathic in CV-1 cells so that all tests based on the cytopathic effect could be carried out. The CV-1 cell line can thus be recommended as an extra-ordinarily suitable substrate for the diagnosis and study of arboviruses.


Cultivation of Tuleniy virus (strain Murman) in vitro

February 1978

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4 Reads

Journal of Hygiene, Epidemiology, Microbiology, and Immunology

In vitro culturability of Murman strain of Tuleniy flavivirus isolated recently in the northern regions of the USSR was studied. Stable PS pig kidney line was found suitable as a primary sensitive cell substrate for the isolation, proliferation and serial propagation of the virus. The pronounced pathogenicity of the virus to PS cells permits the testing of its infective activity comparable with i.c. titrations on mice, VNT in vitro and the plaquing technique. PS line is suitable for the demonstration and identification of the virus antigen and/or for the study of reproduction of the virus on cellular level using the technique of immunofluorescence.



[Practical modifications of media for tissue cultures (author's transl)]

February 1976

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4 Reads

Ceskoslovenská Epidemiologie, Mikrobiologie, Imunologie

A description is given of the technique of preparing pulverized autoclaveable media for tissue cultures in growth and maintenance modifications as well as in those with buffer action in untreated atmosphere. Details are also given of the technique of radiation sterilization of the serum and other non autoclaveable ingredients necessary for in vitro cell cultivation. These technological procedures make it possible to obtain media with exceptionally long expiration times with absolute sterility in all tissue media components. A significant methodological contribution was observed particularly in the virological uses of the respective modifications of media for tissue cultures.


Experiences with the adaptation of Crimean hemorrhagic fever virus to the CV 1 monkey cell line

August 1975

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4 Reads

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4 Citations

Acta Virologica

Of several cell lines tested, the CV-1 monkey cell line proved to be the most suitable system for propagation of Crimean hemorrhagic fever (CHF) virus. Two CHF strains could be adapted to this cell line by serial passaging. The viral progeny from these passages was specifically cytocidal and produced plaques in CV-1 cells, which could be used in serological diagnosis of CHF. After adaptation of CHF virus to CV-1 cells, the rate of virus synthesis was more rapid, but the yield of virus produced by these cells did not substantially increase. Data reported on cultivation of CHF-Congo viruses are discussed in comparison with the present findings.


A Modified Plaque Method for Arboviruses on Plastic Panels

May 1975

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9 Reads

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8 Citations

Acta Virologica

Autoclavable culture media containing an increased (10---15-fold)concentration of succinate buffer permit a comparitively long-term cultivation of cells in free gas exchange with the atmosphere. Based on them, an economical technique of plaque titration of arboviruses on plastic panels with methylcellulose overlay was developed. With seven arboviruses of three different groups and three cell lines (CV-1, PS and PK), the method proved sufficiently sensitive as compared with titrations in mice or tube cell cultures and suitable for plaque reduction tests.


Cultivation of the fixed rabies virus strain HEP Flury in vitro. I. Adaptation of the HEP Flury strain to the human diploid cell line HEL

April 1973

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8 Reads

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5 Citations

Journal of Hygiene, Epidemiology, Microbiology, and Immunology

The authors verified the finding that adaptation of the fixed rabies strain HEP Flury to a line of diploid human embryonic lung cells (HEL) requires prolonged use of the cell-mixing transfer technique (the transfer of mixed suspensions of infected and normal cells). With their modification, the virus was fully adapted from about the 80th-85th passage, when maximum intensity and stabilization of the infection process in the cultures was attained. Once it had been adapted, the virus could be further propagated by transferring infective medium to HEL suspensions or cultures. The CPE, which was observed from the 25th adaptation passage, was characterized by slow progression and did not acquire a focal character until about the 50th passage. The relationship between development of a CPE and the appearance of large inclusions and polymorphous antigen masses in the cytoplasm of infected cells was confirmed.



The dynamics of the formation of virus-neutralizing, complement-fixing and immunofluorescent antibodies to the rabies virus in guinea pigs

February 1972

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2 Reads

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1 Citation

Folia Microbiologica

The aim of the present work was to establish the dynamics of the formation of virus-neutralizing (VN), complement-fixing (CF) and immunofluorescent (IF) antibodies in guinea pig antisera or fractions (IgM, IgG, 7S γ-2 7S γ-1 and F-fraction) obtained by gel filtration on a G-200 Sephadex column, or by chromatography on DEAE cellulose. It was shown that (I) there exists a correlation between the development and titres of VN and IF antibodies. This correlation was observed in both whole serum and its fractions IgG, 7S γ-2 and 7S γ-1 during the whole experimental period. (2) The formation of CF antibodies, followed a different pattern in comparison with VN and IF antibodies. (3) VN, IF and CF antibodies were found to be bound to the IgG fraction, and most of the activity was found in subfraction 7S γ-2. The subfraction 7S γ-1 possessed approximately one half of the activity of fraction 7S γ-2 as regards VN and IF antibodies, and almost no activity of CF antibodies. (4) Both VN and CF antibodies, present in the IgM fraction, reached the maximum in the second blood sample, i.e. 14 days after the first dose of virulent virus. In the further course of the antibody response in guinea pigs, the curve of VN, IF and CF antibodies showed a reversed trend in the whole serum rather than in the IgM fraction.


Citations (10)


... Virus growth and purification. Confluent monolayers of CV-1 cells in 1750 cm 2 plastic roller bottles (Falcon; Becton-Dickinson) were infected at a multiplicity of 0-1 in 10 ml of Plaisner's modified Eagle's medium (Plaisner et al., 1974) supplemented with 2 % FCS and antibiotics as above (maintenance medium). After 60 min adsorption, 150 ml maintenance medium was added and incubation continued at 37 °C. ...

Reference:

Structural and Cell-associated Proteins of Lassa Virus
Tissue culture media with a buffering capacity in a non-conditioned gas phase
  • Citing Article

Acta Virologica

... TBEV has been studied in cell cultures in vitro [281]. CPE and distinct plaques of TBEV are produced in porcine embryonal kidney cell lines PK, PS and SPEV [282][283][284] and in CV-1 monkey cells [285][286][287], which are highly applicable for VNT and PRNT. ...

[Experience with the use of low-viscosity methyl cellulose for plaquing of some arboviruses (author's transl)]
  • Citing Article
  • April 1977

Ceskoslovenská Epidemiologie, Mikrobiologie, Imunologie

... vrrus in,CC ofvariOlis originslwere becOming more numerous (Abelseth 1964,Htbllovsky et .. al .. ,1966~, and others). In:'theco\Jr,se of these adaptations,the mainfea,tures, that were followed: included changes in pathogenic properties for laboratory animals (R ulka et. al. 19.74), the occurrence of a cytopathic effect (CPE), neural pathogenicity (Hronovsky et. al. 1968), and the effects of rabies infection on the host cell. Great attention is also paid to production,of large ,quantities of the virus for the purpose of both human and veterinary use. ,'-' " , This, Study was aimed .at : the kinetics of the replication of a street sU-ainof rabies. yirus !>:f local provenience in. the coune ,of it!! adapt ...

Propagation of street rabies virus in primary cultures of dog kidney cells
  • Citing Article
  • April 1966

Acta Virologica

... e vrrus in,CC ofvariOlis originslwere becOming more numerous (Abelseth 1964,Htbllovsky et .. al .. ,1966~, andothers). In:'theco\Jr,se of these adaptations,the mainfea,tures, that were followed: included changes in pathogenic properties for laboratory animals (R ulka et. al. 19.74), the occurrence of a cytopathic effect (CPE), neural pathogenicity (Hronovsky et. al. 1968), and the effects of rabies infection on the host cell. Great attention is also paid to production,of large ,quantities of the virus for the purpose of both human and veterinary use. ...

Process of adaptation of street rabies virus to dog kidney cell primary cultures
  • Citing Article
  • June 1968

Acta Virologica

... Previous studies investigating intranasal or aerosol infection used the fixed RABV strain CVS and EBLV-2 with different results regarding pathogenicity [55][56][57][58]. But although these results seemed to sometimes contradict each other, it has to be noted that the studies were all designed differently. ...

Development of inhalation rabies infection in suckling guinea pigs
  • Citing Article
  • June 1969

Acta Virologica

... Previous studies investigating intranasal or aerosol infection used the fixed RABV strain CVS and EBLV-2 with different results regarding pathogenicity [55][56][57][58]. But although these results seemed to sometimes contradict each other, it has to be noted that the studies were all designed differently. ...

Experimental inhalation infection of laboratory rodents with rabies virus
  • Citing Article
  • June 1969

Acta Virologica

... Rabies virus has been cultivated in vitro with primary hamster kidney [8,23,33], pig kidney [1], dog kidney [15,20], fetal calf kidney [7], green monkey kidney [41], dog salivary gland [14,15], bat brown fat [2], chick embryo (CE) [24,28,39,41,44], human diploid [18,21,41], mouse embryo [41], rat ganglion [29,30], mouse ganglion [29,30], mouse ependymoma [3] and reptilian cells [6,32] as well as in established cell lines of baby hamster kidney [4,5,18], hamster embryo [16,40], green monkey kidney [42], dog kidney [41], rabbit reticuloendothelium [18,19,26], mouse ependymoma [5], pig Fallopian tube [9] and cold-blooded vertebrate origins [11,37,40]. ...

Cultivation of the fixed rabies virus strain HEP Flury in vitro. I. Adaptation of the HEP Flury strain to the human diploid cell line HEL
  • Citing Article
  • April 1973

Journal of Hygiene, Epidemiology, Microbiology, and Immunology

... Acute TBE can be diagnosed through the detection of IgM antibodies using indirect IFA [296][297][298]. The preparation of hyperimmune mouse ascitic fluids and IFA were described as a useful tool for the identification and serodiagnostics of TBEV [299,300]. For rapid serodiagnosis, ELISA with purified TBEV antigen from infected mouse brains can be used [301,302]. ...

Use of hyperimmune mouse ascitic fluids for arbovirus differentiation by indirect immunofluorescence and conventional serology
  • Citing Article
  • October 1981

Acta Virologica