Uso de células de Aedes albopictus C6/36 na propagação e classificação de arbovírus das famílias Togaviridae, Flaviviridae, Bunyaviridae e Rhabdoviridae

Figueiredo, Luiz Tadeu Moraes

doi:10.1590/S0037-86821990000100003

Resumos

Colônias de células de mosquito Aedes albopictus C6/36 foram infectadas com 23 arbovirus, sendo 19 destes existentes no Brasil, pertencentes às famílias Togaviridae, Flaviviridae, Bunyaviridae e Rhabdoviridae. A Replicação virai foi detectada por imunofluorescência indireta com todos os vírus estudados enquanto que o efeito citopático foi observado durante a infecção por alguns destes. No teste de imunofluorescência indireta utilizou-se fluidos ascíticos imunes de camundongos, específicos para os vírus estudados. A replicação virai caracterizada por grande produção de antígeno recomenda a utilização de células C6/36 na propagação e em tentativas de isolamento desses arbovirus. A técnica de imunofluorescência ofereceu importantes subsídios na classificação e identificação de vírus que replicam nestas células.

Arbovirus; Culturas de células de mosquito; Imunofluorescência Indireta; Isolamento de vírus

C6/36 Aedes albopictus cells were infected w ith Brazilian arbovirus from the families Togaviridae, Flaviviridae, Bunyaviridae and Rhabdoviridae. Replication was obtained with all the studied viruses and cytopathic effect was observed with some. Viral antigen was assayed in C6/36 cell cultures for antigen was assayed in C6/36 cells by an indirect immunofluorescence test using specific mouse immune ascitic fluid. Antigen production was detected in C6/36 cells infected with all the studied viruses. The author recommends the inoculation of C6/36 cell cultures for isolation of virus from the four studied families. The immunofluorescence technique is an important tool for classification and identification of virus growing in C6/36 cells.

Arbovirus; Mosquito tissue culture cells; Indirect imunofluorescence; Virus isolation

ARTIGOS

Uso de células de Aedes albopictus C6/36 na propagação e classificação de arbovírus das famílias Togaviridae, Flaviviridae, Bunyaviridae e Rhabdoviridae

Luiz Tadeu Moraes Figueiredo

^{Endereço para correspondência} Endereço para correspondência: Dr. Luiz Tadeu Moraes Figueiredo. Departamento de Clínica Médica. Facuidade de Medicina de Ribeirão Preto/USP. 14049 Ribeirão Preto, SP, Brasil.

RESUMO

Colônias de células de mosquito Aedes albopictus C6/36 foram infectadas com 23 arbovirus, sendo 19 destes existentes no Brasil, pertencentes às famílias Togaviridae, Flaviviridae, Bunyaviridae e Rhabdoviridae. A Replicação virai foi detectada por imunofluorescência indireta com todos os vírus estudados enquanto que o efeito citopático foi observado durante a infecção por alguns destes. No teste de imunofluorescência indireta utilizou-se fluidos ascíticos imunes de camundongos, específicos para os vírus estudados. A replicação virai caracterizada por grande produção de antígeno recomenda a utilização de células C6/36 na propagação e em tentativas de isolamento desses arbovirus. A técnica de imunofluorescência ofereceu importantes subsídios na classificação e identificação de vírus que replicam nestas células.

Palavras-chave: Arbovirus. Culturas de células de mosquito. Imunofluorescência Indireta. Isolamento de vírus.

ABSTRACT

C6/36 Aedes albopictus cells were infected w ith Brazilian arbovirus from the families Togaviridae, Flaviviridae, Bunyaviridae and Rhabdoviridae. Replication was obtained with all the studied viruses and cytopathic effect was observed with some. Viral antigen was assayed in C6/36 cell cultures for antigen was assayed in C6/36 cells by an indirect immunofluorescence test using specific mouse immune ascitic fluid. Antigen production was detected in C6/36 cells infected with all the studied viruses. The author recommends the inoculation of C6/36 cell cultures for isolation of virus from the four studied families. The immunofluorescence technique is an important tool for classification and identification of virus growing in C6/36 cells.

Keywords: Arbovirus. Mosquito tissue culture cells. Indirect imunofluorescence. Virus isolation.

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Full text available only in PDF format.

Recebido para publicação em 03/08/89.

Trabalho efetuado no Departamento de Virologia do Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brasil. Financiado em parte pelo CNPq (302509/87-9/BM/FV) e pela SUCAM.

1. Brandt WE, Buescher EL, Hetrick FM. Production and characterization of arbovirus antibody in mouse ascitic fluid. The American Journal of Tropical Medicine and Hygiene 16:339-347, 1967.
2. Brown DT, Condreay LD. Replication of alphaviruses in mosquito cells. In: Schlesinger S, Schlesinger MJ. The Togaviridae and Flaviviridae, 1 st edition, Plenum, New York, p. 171-203, 1986.
3. Causey CE, Causey OR Development of arbovirus studies in the Amazon Region. In: International Symposium on Tropical Arboviruses and Haemorrhagic Fevers, Belem, 1980. Academia Brasileira de Ciências, Rio de Janeiro, p. 13-20, 1982.
4. DégallierN, Hervé JP, Rosa AP AT, Vasconcelos PFC, Rosa JFST, Sá Filho GC. A ecologia dos arbovirus na Amazônia: pesquisas atuais e perspectivas. Hiléia Médica 8:1-98, 1987.
5. Figueiredo LT Shope RE. An enzyme immunoassay for dengue antibody using infected cultured mosquito cells as antigen. Journal of Virological Methods 17:191-198, 1987.
6. Gardener PS. Immunofluorescence. In: Specter S, Lancz GJ. Manual of Clinical Virology, 1 st edition, Elsevier, New York, p. 95-109, 1986.
7. Henchal EA, Mc Cown JM, Seguin MC, Gentry MK, Brandt WE. Rapid identification of dengue virus isolates by using monoclonal antibodies in an indirect immunofluorescence assay. The American Journal of Tropical Medicine and Hygiene 32:164-169, 1983.
8. Igarashi A. Isolation of a Singh's Aedes albopictus cell clone sensitive to dengue and Chikungunya viruses. Journal of General Virology 40:531-544, 1978.
9. Karabatsos N (ed). International Catalogue of Arbovirus including certain other viruses of vertebrates. San Antonio, Texas, American Society of Tropical Medicine and Hygiene, 1985.
10. Miller BR, De Foliart GR, Yuill TM. Vertical transmission of La Crosse virus (California encephalitis Group): transovarial and filial infection rates in Aedes triseriatus (Diptera: Culicidae). Journal of Medical Entomology 14:437-440, 1977.
11. Pinheiro FP. Situação das arboviroses na Região Amazônica. In: International Simposium on Tropical Arboviruses and Haemorrhagic Fevers, Belém, 1980. Rio de Janeiro,Academia Brasileira de Ciências, p. 27-48, 1982.
12. Rosen L. Further observations on the mechanism of vertical transmission of flaviviruses by Aedes mosquitoes. The American Journal of Tropical Medicine and Hygiene 39:123-126, 1988.
13. Sakurai T, Bundo K, Igarashi A. Detection of Rocio virus-specific polypeptides in virus-infected cells and culture by western blotting method. Tropical Medicine 27:129-140, 1985.
14. Schatzmayr HG, Nogueira RMR, Rosa APAT. An outbreak of dengue virus at Rio de J aneiro. Memórias do Instituto Oswaldo Cruz 81:245-246, 1986.
15. Shope RE, Sather GE. Arboviruses. In: Lennette FH, Schimidt NJ (ed). Diagnostic Procedures for Viral, Rickettsial and Clamydial Infections, 2nd edition, American Public Health Association, Washington, p. 767- 814, 1979.
16. Singh K.R Propagation of arboviruses in Singh's cell lines. I. Growth of arboviruses in Aedes albopictus and A. aegypti cell lines. Current Topics in Microbiology and Immunology 55:127-133, 1971.
17. Tesh RB. A method for the isolation and identification of dengue viruses, using mosquito cell cultures. The American Journal of Tropical Medicine and Hygiene 28:1053-1059, 1979.
18. Theiler M, Downs WG. Virus classification. In: The arthropodbome viruses of vertebrates. An account of The Rockefeller Foundation Virus Program 1951-1970. Yale Univesity, New Haven, p. 95-316, 1973.
19. Westaway EG. Flavivirus replication strategy. Advances in Virus Research 33:45-90, 1987.

Endereço para correspondência:

Dr. Luiz Tadeu Moraes Figueiredo.

Departamento de Clínica Médica.

Facuidade de Medicina de Ribeirão Preto/USP.

14049

Ribeirão Preto, SP, Brasil.

Datas de Publicação

Publicação nesta coleção
27 Maio 2013
Data do Fascículo
Mar 1990

Histórico

Recebido
03 Ago 1989
Aceito
03 Ago 1989

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. Brandt WE, Buescher EL, Hetrick FM. Production and characterization of arbovirus antibody in mouse ascitic fluid. The American Journal of Tropical Medicine and Hygiene 16:339-347, 1967.

[2] 2. Brown DT, Condreay LD. Replication of alphaviruses in mosquito cells. In: Schlesinger S, Schlesinger MJ. The Togaviridae and Flaviviridae, 1 st edition, Plenum, New York, p. 171-203, 1986.

[3] 3. Causey CE, Causey OR Development of arbovirus studies in the Amazon Region. In: International Symposium on Tropical Arboviruses and Haemorrhagic Fevers, Belem, 1980. Academia Brasileira de Ciências, Rio de Janeiro, p. 13-20, 1982.

[4] 4. DégallierN, Hervé JP, Rosa AP AT, Vasconcelos PFC, Rosa JFST, Sá Filho GC. A ecologia dos arbovirus na Amazônia: pesquisas atuais e perspectivas. Hiléia Médica 8:1-98, 1987.

[5] 5. Figueiredo LT Shope RE. An enzyme immunoassay for dengue antibody using infected cultured mosquito cells as antigen. Journal of Virological Methods 17:191-198, 1987.

[6] 6. Gardener PS. Immunofluorescence. In: Specter S, Lancz GJ. Manual of Clinical Virology, 1 st edition, Elsevier, New York, p. 95-109, 1986.

[7] 7. Henchal EA, Mc Cown JM, Seguin MC, Gentry MK, Brandt WE. Rapid identification of dengue virus isolates by using monoclonal antibodies in an indirect immunofluorescence assay. The American Journal of Tropical Medicine and Hygiene 32:164-169, 1983.

[8] 8. Igarashi A. Isolation of a Singh's Aedes albopictus cell clone sensitive to dengue and Chikungunya viruses. Journal of General Virology 40:531-544, 1978.

[9] 9. Karabatsos N (ed). International Catalogue of Arbovirus including certain other viruses of vertebrates. San Antonio, Texas, American Society of Tropical Medicine and Hygiene, 1985.

[10] 10. Miller BR, De Foliart GR, Yuill TM. Vertical transmission of La Crosse virus (California encephalitis Group): transovarial and filial infection rates in Aedes triseriatus (Diptera: Culicidae). Journal of Medical Entomology 14:437-440, 1977.

[11] 11. Pinheiro FP. Situação das arboviroses na Região Amazônica. In: International Simposium on Tropical Arboviruses and Haemorrhagic Fevers, Belém, 1980. Rio de Janeiro,Academia Brasileira de Ciências, p. 27-48, 1982.

[12] 12. Rosen L. Further observations on the mechanism of vertical transmission of flaviviruses by Aedes mosquitoes. The American Journal of Tropical Medicine and Hygiene 39:123-126, 1988.

[13] 13. Sakurai T, Bundo K, Igarashi A. Detection of Rocio virus-specific polypeptides in virus-infected cells and culture by western blotting method. Tropical Medicine 27:129-140, 1985.

[14] 14. Schatzmayr HG, Nogueira RMR, Rosa APAT. An outbreak of dengue virus at Rio de J aneiro. Memórias do Instituto Oswaldo Cruz 81:245-246, 1986.

[15] 15. Shope RE, Sather GE. Arboviruses. In: Lennette FH, Schimidt NJ (ed). Diagnostic Procedures for Viral, Rickettsial and Clamydial Infections, 2nd edition, American Public Health Association, Washington, p. 767- 814, 1979.

[16] 16. Singh K.R Propagation of arboviruses in Singh's cell lines. I. Growth of arboviruses in Aedes albopictus and A. aegypti cell lines. Current Topics in Microbiology and Immunology 55:127-133, 1971.

[17] 17. Tesh RB. A method for the isolation and identification of dengue viruses, using mosquito cell cultures. The American Journal of Tropical Medicine and Hygiene 28:1053-1059, 1979.

[18] 18. Theiler M, Downs WG. Virus classification. In: The arthropodbome viruses of vertebrates. An account of The Rockefeller Foundation Virus Program 1951-1970. Yale Univesity, New Haven, p. 95-316, 1973.

[19] 19. Westaway EG. Flavivirus replication strategy. Advances in Virus Research 33:45-90, 1987.