AEGY-28 Cell Line of Aedes aegypti (Diptera Culicidae) is Infection Refractory to Dengue 2 and Yellow Fever Virus
Mosquito cell derived cultures are useful tools for arbovirus isolation, identification or characterization. For studying dengue (DENV) and yellow fever viruses (YFV) Aedes albopictus C6/36 or Aedes pseudoscutellaris AP-61 cell lines, are normally used. The Aedes aegypti AEGY-28 cell line was obtained from embryonic tissues and characterized previously by one of us. In order to evaluate its susceptibility to two Flavivirus, AEGY- 28 cells were inoculated with different multiplicity of infection (MOI) with type 2 DENV (COL-789, MOI: 1 and 5) and YFV clinical isolates (V-341, MOI 0,02) then processed at different times post infection (p.i.). Immunostai ning and fluorometric cell-ELISA were carried out to identify and quantify viral antigens. C6/36 and Vero cells were used as positive controls. Unexpectedly, immunoreactivity was not found in inoculated AEGY-28 cells, even in higher MOI or late times p.i., therefore antigen quantification using fluorometric cell-ELISA were not plausible. Reverse transcriptase PCR with specific primers did not detect viral RNA in AEGY-28 inoculated cells. We can conclude that Aedes aegypti AEGY-28 cell line is not susceptible to dengue and yellow fever Flavivirus, a finding possibly related with the lacking of specific molecules at the plasma membrane or absence of cell machinery necessary for viral replication.
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Universidad Nacional de Colombia - Sede Bogotá - Facultad de Ciencias - Departamento de Biología
2007
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oai:www.revistas.unal.edu.co:article-271932014-06-08T04:09:21Z AEGY-28 Cell Line of Aedes aegypti (Diptera Culicidae) is Infection Refractory to Dengue 2 and Yellow Fever Virus Linea celular de Aedes aegypti (Diptera Culicidae) AEGY-28 refractaria a la infección con los virus dengue 2 y fiebre amarilla Castañeda, Nadia Y. Castellanos, Jaime E. Zapata, Angela C. Bello, Felio Flavivirus Cell Line Dengue Virus Yellow Fever Arboviruses Infection Flavivirus Línea Celular Virus Dengue Fiebre Amarilla Arbovirus Infección Mosquito cell derived cultures are useful tools for arbovirus isolation, identification or characterization. For studying dengue (DENV) and yellow fever viruses (YFV) Aedes albopictus C6/36 or Aedes pseudoscutellaris AP-61 cell lines, are normally used. The Aedes aegypti AEGY-28 cell line was obtained from embryonic tissues and characterized previously by one of us. In order to evaluate its susceptibility to two Flavivirus, AEGY- 28 cells were inoculated with different multiplicity of infection (MOI) with type 2 DENV (COL-789, MOI: 1 and 5) and YFV clinical isolates (V-341, MOI 0,02) then processed at different times post infection (p.i.). Immunostai ning and fluorometric cell-ELISA were carried out to identify and quantify viral antigens. C6/36 and Vero cells were used as positive controls. Unexpectedly, immunoreactivity was not found in inoculated AEGY-28 cells, even in higher MOI or late times p.i., therefore antigen quantification using fluorometric cell-ELISA were not plausible. Reverse transcriptase PCR with specific primers did not detect viral RNA in AEGY-28 inoculated cells. We can conclude that Aedes aegypti AEGY-28 cell line is not susceptible to dengue and yellow fever Flavivirus, a finding possibly related with the lacking of specific molecules at the plasma membrane or absence of cell machinery necessary for viral replication. Los cultivos celulares de mosquitos son frecuentemente utilizados para el aislamiento, identificación y caracterización de arbovirus. Para el estudio de los virus dengue (VDEN) y virus de fiebre amarilla (VFA) se emplean, principalmente, la línea celular C6/36 de Aedes albopictus y la línea celular AP-61, obtenida de Aedes pseudoscutellaris. La línea celular de A. aegypti AEGY-28, previamente obtenida a partir de tejidos embrionarios del vector, se utilizó en el presente trabajo para evaluar la susceptibilidad a la infección por VDEN y VFA. Para ello, los cultivos celulares se ensayaron a diferente multiplicidad de infección con los aislados clínicos de virus dengue tipo 2 (COL-789, MOI: 1 y 5) y VFA (V-341, MOI 0,02). Posteriormente se realizó la detección de antígenos virales por la técnica de inmunocitoquímica y su cuantificación por la técnica de Cell-ELISA fluorométrica. Se usaron como controles positivos de infección, tanto células C6/36 como células VERO. Inesperadamente, no se observó inmunoreactividaden las células de A. aegypti infectadas con ambos tipos de virus en ninguno de los MOI o tiempos estudiados. Tampoco se evidenció antígeno por la técnica fluorométrica ni fue posible detectar RNA viral por RT-PCR a partir de células infectadas.Por lo tanto, se puede concluir que la línea celular de A. aegypti no es susceptible a la infección por VDEN ni por VFA. Ello podría estar relacionado con características propias de la membrana celular o de la maquinaria enzimática necesaria para la replicación viral. Universidad Nacional de Colombia - Sede Bogotá - Facultad de Ciencias - Departamento de Biología 2007-07-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://revistas.unal.edu.co/index.php/actabiol/article/view/27193 Acta Biológica Colombiana; Vol. 12 No. 2 (2007); 47-57 Acta Biológica Colombiana; Vol. 12 Núm. 2 (2007); 47-57 1900-1649 0120-548X spa https://revistas.unal.edu.co/index.php/actabiol/article/view/27193/27466 Derechos de autor 2007 Acta Biológica Colombiana https://creativecommons.org/licenses/by/4.0 |
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Castañeda, Nadia Y. Castellanos, Jaime E. Zapata, Angela C. Bello, Felio |
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Castañeda, Nadia Y. Castellanos, Jaime E. Zapata, Angela C. Bello, Felio AEGY-28 Cell Line of Aedes aegypti (Diptera Culicidae) is Infection Refractory to Dengue 2 and Yellow Fever Virus |
author_facet |
Castañeda, Nadia Y. Castellanos, Jaime E. Zapata, Angela C. Bello, Felio |
author_sort |
Castañeda, Nadia Y. |
title |
AEGY-28 Cell Line of Aedes aegypti (Diptera Culicidae) is Infection Refractory to Dengue 2 and Yellow Fever Virus |
title_short |
AEGY-28 Cell Line of Aedes aegypti (Diptera Culicidae) is Infection Refractory to Dengue 2 and Yellow Fever Virus |
title_full |
AEGY-28 Cell Line of Aedes aegypti (Diptera Culicidae) is Infection Refractory to Dengue 2 and Yellow Fever Virus |
title_fullStr |
AEGY-28 Cell Line of Aedes aegypti (Diptera Culicidae) is Infection Refractory to Dengue 2 and Yellow Fever Virus |
title_full_unstemmed |
AEGY-28 Cell Line of Aedes aegypti (Diptera Culicidae) is Infection Refractory to Dengue 2 and Yellow Fever Virus |
title_sort |
aegy-28 cell line of aedes aegypti (diptera culicidae) is infection refractory to dengue 2 and yellow fever virus |
description |
Mosquito cell derived cultures are useful tools for arbovirus isolation, identification or characterization. For studying dengue (DENV) and yellow fever viruses (YFV) Aedes albopictus C6/36 or Aedes pseudoscutellaris AP-61 cell lines, are normally used. The Aedes aegypti AEGY-28 cell line was obtained from embryonic tissues and characterized previously by one of us. In order to evaluate its susceptibility to two Flavivirus, AEGY- 28 cells were inoculated with different multiplicity of infection (MOI) with type 2 DENV (COL-789, MOI: 1 and 5) and YFV clinical isolates (V-341, MOI 0,02) then processed at different times post infection (p.i.). Immunostai ning and fluorometric cell-ELISA were carried out to identify and quantify viral antigens. C6/36 and Vero cells were used as positive controls. Unexpectedly, immunoreactivity was not found in inoculated AEGY-28 cells, even in higher MOI or late times p.i., therefore antigen quantification using fluorometric cell-ELISA were not plausible. Reverse transcriptase PCR with specific primers did not detect viral RNA in AEGY-28 inoculated cells. We can conclude that Aedes aegypti AEGY-28 cell line is not susceptible to dengue and yellow fever Flavivirus, a finding possibly related with the lacking of specific molecules at the plasma membrane or absence of cell machinery necessary for viral replication. |
publisher |
Universidad Nacional de Colombia - Sede Bogotá - Facultad de Ciencias - Departamento de Biología |
publishDate |
2007 |
url |
https://revistas.unal.edu.co/index.php/actabiol/article/view/27193 |
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