In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS
Descripción del Articulo
The aim of the study was to determine the relative in vitro ARNm expression levels of IL-2 and IL-10 in peripheral blood leukocytes by real time RT-PCR and relative quantification using the 2-ΔΔCt method in presence of clostridial antigens and GAPDH as an endogenous control. Whole blood was collecte...
| Autores: | , , |
|---|---|
| Formato: | artículo |
| Fecha de Publicación: | 2014 |
| Institución: | Universidad Nacional Mayor de San Marcos |
| Repositorio: | Revistas - Universidad Nacional Mayor de San Marcos |
| Lenguaje: | español |
| OAI Identifier: | oai:ojs.csi.unmsm:article/10121 |
| Enlace del recurso: | https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/10121 |
| Nivel de acceso: | acceso abierto |
| Materia: | interleukin-2 interleukin-10 Clostridium perfringens real time RT-PCR relative quantification interleucina 2 interleucina 10 RT-PCR tiempo real cuantificación relativa |
| id |
REVUNMSM_fd2cbe7ba96437edcfb3cc265f341077 |
|---|---|
| oai_identifier_str |
oai:ojs.csi.unmsm:article/10121 |
| network_acronym_str |
REVUNMSM |
| network_name_str |
Revistas - Universidad Nacional Mayor de San Marcos |
| repository_id_str |
|
| spelling |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENSEXPRESIÓN in vitro DE LAS INTERLEUCINAS 2 Y 10 DE LINFOCITOS DE ALPACAS (Vicugna pacos) EN PRESENCIA DE ANTÍGENOS CLOSTRIDIALESWatanabe W., RaquelManchego S., AlbertoRivera G., Hermelindainterleukin-2interleukin-10Clostridium perfringensreal time RT-PCRrelative quantificationinterleucina 2interleucina 10Clostridium perfringensRT-PCR tiempo realcuantificación relativaThe aim of the study was to determine the relative in vitro ARNm expression levels of IL-2 and IL-10 in peripheral blood leukocytes by real time RT-PCR and relative quantification using the 2-ΔΔCt method in presence of clostridial antigens and GAPDH as an endogenous control. Whole blood was collected from 10 adult alpacas. Samples were centrifuged to obtain leukocytes using the Ficoll reagent, purified with ammonium chloride and cultured in 24-well plates at a concentration of 500 000 live cells/ml minimal essential medium (MEM) with clostridial extract suspensions at concentrations of 400, 16, 0.8 and 0.2 µg/ml. Subsequently, the RT-qPCR test was performed. IL-2 kinetic expression patterns were inversely proportional to the clostridial antigen dose in the three incubation times (p>0.05), unlike IL-10 which its kinetic expression patterns were variable, reaching its maximum at 12 h (p<0.05) while showing a similar trend like IL-2 in the 16 µg/ml clostridial antigen dose. IL-10 expression exceeded 40-fold the control expression respect to IL-2. A strong polarization towards the Th2 subtype was shown.El objetivo del presente estudio fue determinar, mediante la técnica de RT-PCR Tiem- po Real y cuantificación relativa según el método 2-ΔΔCt, los niveles relativos de expresión in vitro de ARN mensajero de IL-2 e IL-10 en leucocitos circulantes de alpacas en presencia de antígenos clostridiales, empleándose como control endógeno a GAPDH. Se colectó sangre entera de 10 alpacas adultas. Las muestras fueron centrifugadas para obtener la fracción leucocitaria en gradiente de Ficoll, colectándose la capa flogística. Se hizo un pool de leucocitos que fueron purificados con cloruro de amonio. Se centrifugó y la concentración se ajustó a 500 000 células vivas/ml de medio mínimo esencial (MEM). Fueron cultivados en placas para cultivo celular de 24 pocillos y enfrentados a suspensiones de extracto clostridial a las concentraciones de 400, 16, 0.8 y 0.2 µg/ml por 1, 12 y 24 h. Posteriormente, se realizó la RT-qPCR. La cinética de la expresión de IL-2 mostró una tendencia no significativa inversamente proporcional a la dosis de antígeno clostridial en los tres tiempos de incubación, a diferencia de IL-10 cuya expresión fue variable, alcan- zando el máximo a las 12 h de incubación (p<0.05) y mostrando un patrón similar al de IL-2 en la dosis de 16 µg/ml de antígeno clostridial. La expresión de IL-10 superó hasta en 40 veces lo expresado por el calibrador respecto a IL-2, evidenciándose una marcada respuesta hacia la polarización del subtipo Th2.Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria2014-09-15info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/1012110.15381/rivep.v25i3.10121Revista de Investigaciones Veterinarias del Perú; Vol. 25 Núm. 3 (2014); 419-429Revista de Investigaciones Veterinarias del Perú; Vol. 25 No. 3 (2014); 419-4291682-34191609-9117reponame:Revistas - Universidad Nacional Mayor de San Marcosinstname:Universidad Nacional Mayor de San Marcosinstacron:UNMSMspahttps://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/10121/8861Derechos de autor 2014 Raquel Watanabe W., Alberto Manchego S., Hermelinda Rivera G.https://creativecommons.org/licenses/by-nc-sa/4.0info:eu-repo/semantics/openAccessoai:ojs.csi.unmsm:article/101212020-03-29T18:08:50Z |
| dc.title.none.fl_str_mv |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS EXPRESIÓN in vitro DE LAS INTERLEUCINAS 2 Y 10 DE LINFOCITOS DE ALPACAS (Vicugna pacos) EN PRESENCIA DE ANTÍGENOS CLOSTRIDIALES |
| title |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS |
| spellingShingle |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS Watanabe W., Raquel interleukin-2 interleukin-10 Clostridium perfringens real time RT-PCR relative quantification interleucina 2 interleucina 10 Clostridium perfringens RT-PCR tiempo real cuantificación relativa |
| title_short |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS |
| title_full |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS |
| title_fullStr |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS |
| title_full_unstemmed |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS |
| title_sort |
In vitro EXPRESSION OF INTERLEUKIN-2 AND -10 FROM ALPACA (Vicugna pacos) LEUKOCYTES IN PRESENCE OF CLOSTRIDIAL ANTIGENS |
| dc.creator.none.fl_str_mv |
Watanabe W., Raquel Manchego S., Alberto Rivera G., Hermelinda |
| author |
Watanabe W., Raquel |
| author_facet |
Watanabe W., Raquel Manchego S., Alberto Rivera G., Hermelinda |
| author_role |
author |
| author2 |
Manchego S., Alberto Rivera G., Hermelinda |
| author2_role |
author author |
| dc.subject.none.fl_str_mv |
interleukin-2 interleukin-10 Clostridium perfringens real time RT-PCR relative quantification interleucina 2 interleucina 10 Clostridium perfringens RT-PCR tiempo real cuantificación relativa |
| topic |
interleukin-2 interleukin-10 Clostridium perfringens real time RT-PCR relative quantification interleucina 2 interleucina 10 Clostridium perfringens RT-PCR tiempo real cuantificación relativa |
| description |
The aim of the study was to determine the relative in vitro ARNm expression levels of IL-2 and IL-10 in peripheral blood leukocytes by real time RT-PCR and relative quantification using the 2-ΔΔCt method in presence of clostridial antigens and GAPDH as an endogenous control. Whole blood was collected from 10 adult alpacas. Samples were centrifuged to obtain leukocytes using the Ficoll reagent, purified with ammonium chloride and cultured in 24-well plates at a concentration of 500 000 live cells/ml minimal essential medium (MEM) with clostridial extract suspensions at concentrations of 400, 16, 0.8 and 0.2 µg/ml. Subsequently, the RT-qPCR test was performed. IL-2 kinetic expression patterns were inversely proportional to the clostridial antigen dose in the three incubation times (p>0.05), unlike IL-10 which its kinetic expression patterns were variable, reaching its maximum at 12 h (p<0.05) while showing a similar trend like IL-2 in the 16 µg/ml clostridial antigen dose. IL-10 expression exceeded 40-fold the control expression respect to IL-2. A strong polarization towards the Th2 subtype was shown. |
| publishDate |
2014 |
| dc.date.none.fl_str_mv |
2014-09-15 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/10121 10.15381/rivep.v25i3.10121 |
| url |
https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/10121 |
| identifier_str_mv |
10.15381/rivep.v25i3.10121 |
| dc.language.none.fl_str_mv |
spa |
| language |
spa |
| dc.relation.none.fl_str_mv |
https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/10121/8861 |
| dc.rights.none.fl_str_mv |
Derechos de autor 2014 Raquel Watanabe W., Alberto Manchego S., Hermelinda Rivera G. https://creativecommons.org/licenses/by-nc-sa/4.0 info:eu-repo/semantics/openAccess |
| rights_invalid_str_mv |
Derechos de autor 2014 Raquel Watanabe W., Alberto Manchego S., Hermelinda Rivera G. https://creativecommons.org/licenses/by-nc-sa/4.0 |
| eu_rights_str_mv |
openAccess |
| dc.format.none.fl_str_mv |
application/pdf |
| dc.publisher.none.fl_str_mv |
Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria |
| publisher.none.fl_str_mv |
Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria |
| dc.source.none.fl_str_mv |
Revista de Investigaciones Veterinarias del Perú; Vol. 25 Núm. 3 (2014); 419-429 Revista de Investigaciones Veterinarias del Perú; Vol. 25 No. 3 (2014); 419-429 1682-3419 1609-9117 reponame:Revistas - Universidad Nacional Mayor de San Marcos instname:Universidad Nacional Mayor de San Marcos instacron:UNMSM |
| instname_str |
Universidad Nacional Mayor de San Marcos |
| instacron_str |
UNMSM |
| institution |
UNMSM |
| reponame_str |
Revistas - Universidad Nacional Mayor de San Marcos |
| collection |
Revistas - Universidad Nacional Mayor de San Marcos |
| repository.name.fl_str_mv |
|
| repository.mail.fl_str_mv |
|
| _version_ |
1795238225730600960 |
| score |
13.914502 |
Nota importante:
La información contenida en este registro es de entera responsabilidad de la institución que gestiona el repositorio institucional donde esta contenido este documento o set de datos. El CONCYTEC no se hace responsable por los contenidos (publicaciones y/o datos) accesibles a través del Repositorio Nacional Digital de Ciencia, Tecnología e Innovación de Acceso Abierto (ALICIA).
La información contenida en este registro es de entera responsabilidad de la institución que gestiona el repositorio institucional donde esta contenido este documento o set de datos. El CONCYTEC no se hace responsable por los contenidos (publicaciones y/o datos) accesibles a través del Repositorio Nacional Digital de Ciencia, Tecnología e Innovación de Acceso Abierto (ALICIA).
