Kinetics of cytokine expression in bovine PBMCs and whole blood after in vitro stimulation with foot-and-mouth disease virus (FMDV) antigen

Dar, Pervaiz Ahmad; Hajam, Irshad Ahmed; Suryanarayana, Velavurthy S.; Kishore, Subodh; Kondabattula, Ganesh

doi:10.1016/j.cyto.2014.12.011

Cited by 14 publications

(18 citation statements)

References 19 publications

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“…The amount of MTT formazan produced during the incubation was measured by an ELISA reader (Bio-Tek ELx800) at a test wavelength of 550 nm and a reference wavelength of 630 nm. The results were calculated on the optical density and expressed as a stimulation index (SI) (11,15 The amplification reactions were carried out in a final volume of 20 µl as previously described (16,17). The thermal profile used for the reactions of cytokines was to heat samples for 1 min at 95 °C as the initial denaturation program; followed by 50 concurrent cycles involving 94 °C for 1 sec as denaturation program; 54 °C for 5 sec for IL-4 and 52 °C for 5 sec for IFN- and 56 °C for 20 sec for GAPDH as annealing program; 72 °C for 10 sec as extension program; and finally cooling to 40 °C as the cooling program.…”

Section: Virus Preparationmentioning

confidence: 99%

“…The thermal profile used for the reactions of cytokines was to heat samples for 1 min at 95 °C as the initial denaturation program; followed by 50 concurrent cycles involving 94 °C for 1 sec as denaturation program; 54 °C for 5 sec for IL-4 and 52 °C for 5 sec for IFN- and 56 °C for 20 sec for GAPDH as annealing program; 72 °C for 10 sec as extension program; and finally cooling to 40 °C as the cooling program. Gene expression was determined as fold increase given by the 2 -CT formula (16)(17)(18).…”

Section: Virus Preparationmentioning

confidence: 99%

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Evaluation of Cell-mediated Immune Response in PBMCs of Calves Vaccinated by Capri Pox Vaccines Using ELISA and Real-time RT-PCR

et al. 2017

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Background: The analysis of antigen-specific cytokine expression has been considered to evaluate the immune responses and vaccines efficacy in recent years. The aim of this study was to compare the cell-mediated immune response characteristics of two Capri pox virus (CaPV) vaccines against lumpy skin disease in cattle. Materials and Methods: Two Capri pox virus vaccines were administered to dairy cows of two farms and followed up to 5 weeks post vaccination. These vaccines were live attenuated Goat pox virus (GTP) Gorgan strain (n=20) and Sheep pox virus (SPP) Romanian strain (n=20). Cell-mediated immune response of vaccinated calves was evaluated using in vitro lymphocyte proliferation and IFN- and IL-4 release assay after stimulation with recall vaccine strains, and in vivo cytokine expression in PBMCs by real-time PCR. Results: Lymphocyte proliferation in GTP-and SPP-vaccinated groups began to increase till reached to its peak at third week post vaccination and then decreased in the weeks thereafter. Stimulation index in stimulated PBMCs in GTPvaccinated calves was higher than SPP-vaccinated calves in all weeks, which indicated higher levels of immunogenicity produced by the GTP-vaccine in cattle. Also, in both vaccinated groups the peak release of IFN- and IL-4 proteins in cultured PBMCs in response to recall antigen was detected at week 3 post vaccination. Although the mean of the cytokine release in GTP-vaccinated calves was higher than SPP-vaccinated calves in all weeks of experiment, a significant difference was only observed at week 3 post vaccination (P<0.05). In contrast, the IFN- mRNA expression in PBMCs of vaccinated groups was induced early, peaked at week 1 post vaccination and decreased in the weeks thereafter, and this rate was higher in GTP-vaccinated calves compared with SPP-vaccinated calves in all weeks, but the significant difference was only found at week 3 post vaccination (P<0.05). However, the IL-4 mRNA expression showed delayed induction and peaked at week 3, and unlike the SPP group, it remained at this level in GTP group, until the end of experiment. Also this rate of expression in GTP-vaccinated calves was higher than SPP-vaccinated calves in all weeks and had a significant increase at week 5 post vaccination (P<0.05). Conclusion:The findings show that due to induction of high level cell-mediated immune response in live attenuated GTP vaccine compared to SPP vaccine, GTP vaccine has a good immunogenic response, and therefore can be a better choice for vaccination against lumpy skin disease.

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Section: Virus Preparationmentioning

confidence: 99%

Section: Virus Preparationmentioning

confidence: 99%

Evaluation of Cell-mediated Immune Response in PBMCs of Calves Vaccinated by Capri Pox Vaccines Using ELISA and Real-time RT-PCR

et al. 2017

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“…Upon PBMCs activation, coordinated secretion of various cytokines control proliferation, differentiation, and function of cells and are crucial for regulation of immune response (Corry et al 1996;Fan et al 1998;Dar et al 2015). Our results depict significant increase in all the cytokines genes' expression assayed except IL-2, within 1 to 3 h of PHA-P stimulation.…”

Section: Discussionmentioning

confidence: 62%

Dynamics of cytokine gene expression in peripheral blood mononuclear cells of indigenous and exotic breeds of pigs in India

Kirthika

Ali

Behera

et al. 2017

Animal Science Journal

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To incorporate immune competence traits in swine breeding programs, association between immune responsiveness and susceptibility to specific infectious diseases must be established. In order to understand the differences in immune competence between indigenous (Zovawk) and exotic (Large White Yorkshire: LWY) pigs reared in India, we carried out a time course expression analysis of immune-regulating key cytokine genes (interleukin (IL)-2, interferon (IFN)-γ, IL-4 and IL-10) in the phytohemagglutinin-P stimulated peripheral blood mononuclear cells (PBMCs). The IL-2 transcript levels in PBMCs increased several thousand-fold when compared to unstimulated cells in both the breeds, albeit the response in that of Zovawk was remarkably higher. Higher and earlier IFN-γ and IL-4 expression levels in Zovawk pigs suggest that both T 1 and T 2 immune responsiveness of this indigenous breed affords better preparedness for danger signals. Moreover, the low expression levels of IL-10 depict a regulated adaptive immune responsiveness. Remarkable difference between the two breeds of the pigs is evident showing a clear advantage of the Zovawk over LWY in terms of a shorter lag period of adaptive immune response. These findings provide a lead for understanding the genetic differences with respect to immune competence levels of indigenous pigs compared to exotic counterparts.

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“…En un estudio sobre estimulación citocínica de PBMC bovinas frente a un antígeno de una vacuna inactivada del virus de la fiebre aftosa se evaluó el perfil citocínico de IL-2, IL-4 e IFN-γ, observándose altos niveles de expresión a las seis horas en IL-2 e IFN-γ, en tanto que IL-4 llegó a su máxima expresión a las 24 horas de la inducción, encontrando que su perfil citocínico tiene una tendencia al perfil Th1 (Dar et al, 2015). Esto difiere a los hallazgos encontrados en este estudio donde la mayoría de picos de expresión se obtienen a las 48 horas.…”

Section: Discussionunclassified

Evaluación de la inmunogenicidad de una proteína recombinante de una Pasteurella multocida aislada de alpacas con neumonía

Maximiliano

Maturrano

Luna

et al. 2018

Rev. investig. vet. Perú

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El objetivo del estudio fue evaluar la actividad inmunogénica in vitro de una proteína P6-like recombinante procedente de un cultivo de Pasteurella multocida aislada de cuadros de neumonía en crías de alpacas. Se desafiaron cultivos de células mononucleares periféricas sanguíneas (PBMC) con una proteína recombinante P6-like a una concentración de 10 ng por muestra, desde las 3 hasta las 72 horas. Se extrajo ARN totales para realizar la prueba de RT-PCR en tiempo real para observar los niveles de expresión de citoquinas de la respuesta inmune Th1 (TNF-á, IFN-g e IL-2) y Th2 (IL-10 e IL-4) en los tiempos establecidos. Se encontró que tanto las citoquinas con perfil Th1 como Th2 expresan un mayor número de veces con respecto a PBMC no expuestos a la proteína recombinante, siendo las 24 y 48 horas los momentos de mayor expresión. Asimismo, se encontró una aparente tendencia hacia el perfil Th2, pero en niveles que no influyen en la expresión de citoquinas del otro perfil.

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Kinetics of cytokine expression in bovine PBMCs and whole blood after in vitro stimulation with foot-and-mouth disease virus (FMDV) antigen

Cited by 14 publications

References 19 publications

Evaluation of Cell-mediated Immune Response in PBMCs of Calves Vaccinated by Capri Pox Vaccines Using ELISA and Real-time RT-PCR

Evaluation of Cell-mediated Immune Response in PBMCs of Calves Vaccinated by Capri Pox Vaccines Using ELISA and Real-time RT-PCR

Dynamics of cytokine gene expression in peripheral blood mononuclear cells of indigenous and exotic breeds of pigs in India

Evaluación de la inmunogenicidad de una proteína recombinante de una Pasteurella multocida aislada de alpacas con neumonía

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