Volume 5, Issue 2 (May 2017)
Res Mol Med (RMM) 2017, 5(2): 3-8 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Norian R, Afzal ahangaran N, Varshovi H R, Azadmehr A. Evaluation of Cell-mediated Immune Response in PBMCs of Calves Vaccinated by Capri Pox Vaccines Using ELISA and Real-time RT-PCR. Res Mol Med (RMM) 2017; 5 (2) :3-8
URL: http://rmm.mazums.ac.ir/article-1-238-en.html
URL: http://rmm.mazums.ac.ir/article-1-238-en.html
1- Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran
2- Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran ,n.a.ahangran@gmail.com
3- Department of Animal Viral Vaccine, Razi Vaccine and Serum Research Institute, Karaj, Iran
4- Department of Immunology, Babol University of Medical Sciences, Babol, Iran.
2- Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran ,
3- Department of Animal Viral Vaccine, Razi Vaccine and Serum Research Institute, Karaj, Iran
4- Department of Immunology, Babol University of Medical Sciences, Babol, Iran.
Abstract: (8628 Views)
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-g 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 GTP-vaccinated 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-g 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-g 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.
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-g 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 GTP-vaccinated 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-g 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-g 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.
References
1. Tulman ER, Afonso CL, Lu Z, Zsak L, Sur JH, Sandybaev NT, et al. The Genomes of Sheeppox and Goatpox Viruses. J Virol. 2002; 76(12):6054-61. PMID: 12021338
2. Kitching RP. Vaccines for lumpy skin disease, sheep pox and goat pox. Dev Biol (Basel). 2003; 114:161-7. PMID: 14677686
3. Kitching RP, Hammond JM, Taylor WP. A single vaccine for the control of capripox infection in sheep and goats. Res vet sci. 1987; 42(1):53-60. PMID: 3029848
4. Davies FG, Otema C. Relationships of capripox viruses found in Kenya with two Middle Eastern strains and some orthopox viruses. Res vet sci. 1981; 31(2):253-5. PMID: 7034101
5. Barnard AL, Arriens A, Cox S, Barnett P, Kristensen B, Summerfield A, et al. Immune response characteristics following emergency vaccination of pigs against foot-and-mouth disease. Vaccine. 2005; 23(8):1037-47. PMID: 15620477
6. Eble PL, de Bruin MG, Bouma A, van Hemert-Kluitenberg F, Dekker A. Comparison of immune responses after intra-typic heterologous and homologous vaccination against foot-and-mouth disease virus infection in pigs. Vaccine. 2006; 24(9):1274-81. PMID: 16289709
7. Varshovi HR, Keyvanfar H, Aghaiypour K, Pourbakhsh SA, Shooshtari AH, Aghaebrahimian M. Capripoxvirus identification by PCR based on P32 gene. Archives of Razi Institute. 2009; 64(No.1):19-25.
8. OIE. Lumpy skin diseas; Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. paris: World Organization for Animal Health; 2010. p. 1-13.
9. Reed LJ, Muench H. A simple method of estimating fifty per cent endpoints. Am J Epidemiol. 1938; 27(3):493-7.
10. Chrysikopoulos CV, Manariotis ID, Syngouna VI. Virus inactivation by high frequency ultrasound in combination with visible light. Colloids Surf B Biointerfaces. 2013; 107:174-9. PMID: 23500728
11. Delirezh N, Norian R, Azadmehr A. Changes in some pro-and anti-inflammatory cytokines produced by bovine peripheral blood mononuclear cells following foot and mouth disease vaccination. Arch Razi Institute. 2016; 71(3):199-207.
12. Kondo T, Sugiura T, Kamada M, Imagawa H. Colorimetric Assay of Equine Peripheral Lymphocyte Blastogenesis Using MTT. J Equine Sci. 1996; 7(3):63-6.
13. Norian R, Delirezh N, Azadmehr A. Evaluation of proliferation and cytokines production by mitogen-stimulated bovine peripheral blood mononuclear cells. Vet Res Forum. 2015; 6(4):265-71. PMID: 26973760
14. Katial RK, Sachanandani D, Pinney C, Lieberman MM. Cytokine production in cell culture by peripheral blood mononuclear cells from immunocompetent hosts. Clin Diagn Lab immunol. 1998; 5(1):78-81. PMID: 9455885
15. Norian R, Delirezh N, Azadmehr A. Evaluation of proliferation and cytokines production by mitogen-stimulated bovine peripheral blood mononuclear cells. Vet Res Forum. 2015; 6(4):265-71. PMID: 26973760
16. Dar PA, Hajam IA, Suryanarayana VS, Kishore S, Kondabattula G. Kinetics of cytokine expression in bovine PBMCs and whole blood after in vitro stimulation with foot-and-mouth disease virus (FMDV) antigen. Cytokine. 2015; 72(1):58-62. PMID: 25574771
17. Konnai S, Usui T, Ohashi K, Onuma M. The rapid quantitative analysis of bovine cytokine genes by real-time RT-PCR. Vet Microbiol. 2003; 94(4):283-94. PMID: 12829382
18. Zaros LG, Bricarello PA, Amarante AFT, Coutinho LL. Quantification of bovine cytokine gene expression using real-time RT-PCR methodology. Genet Mol Bio. 2007; 30:575-9.
19. Blanco E, Garcia-Briones M, Sanz-Parra A, Gomes P, De Oliveira E, Valero Ml, Et Al. Identification of T-Cell Epitopes in Nonstructural Proteins of Foot-and-Mouth Disease Virus. J Virol. 2001; 75(7):3164-74. PMID: 11238843
20. Saiz JC, Rodriguez A, Gonzalez M, Alonso F, Sobrino F. Heterotypic lymphoproliferative response in pigs vaccinated with foot-and-mouth disease virus. Involvement of isolated capsid proteins. J Gen Virol. 1992; 73 (Pt 10):2601-07. PMID: 1328475
21. Mohamed G, Abdelwahab HAK, Abdelmoneim M, Moustafa Mohamed A, Saad AA. Evaluation of Humoral and Cell-mediated Immunity of Lumpy Skin Disease Vaccine Prepared from Local strainin calves and Its Related to Maternal Immunity. J Am Sci. 2016; 21(10).
22. Heba A, Khafagy MGA, Abdelmoneim M, Mustafa Mohamed A, Saad AA. Preparation and field evaluationof live attenuated sheep pox vaccine for protection of calves against lumpy skin disease. Ben Vet Med J. 2016; 31(2):1-7.
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC-By-NC), which permits use, distribution, and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
Contact Information
