Atlarda DUVAXYN® IE-T Aşısı ile Yapılan Güvenlik Testinden Sonraki İmmunolojik ve Yangısal Reaksiyonlar

Öz

-

Anahtar Kelimeler

• Sitokinler, at, interleukin, qRT-PCR, aşı.

Immunologic and Inflammatory Reactions after Safety Trial in Horses with DUVAXYNIE®-T Vaccine

Öz

Aşı güvenlik testlerinde arasıra beklenmeyen bir ateş ortaya çıkmaktadır. Bu çalışmanın amacı, bu ateş cevabının, inaktive edilmiş virus kompenetleriylemi ilişkili ya da bu cevaba adjuvantın neden olup olmadığını araştırmaktır. On adet immune edilmemiş tay rastgele Grup 1 ve Grup 2 ye ayrılmıştır. Grup 1’in atları (n=5) inaktive edilmiş influenza–tetanus aşısı ile (Duvaxyn®IE-T) intramuskuler olarak aşılanmıştır. Grup 2’nin atları ise carbomer ve aluminyumhidroksid adjuvantı içeren 'Yapmacık' (Sham) aşı ile aşılanmıştır. Rektal ısı test süresi boyunca ölçülmüştür. Atlardan İlk ve ikinci aşılama sonrası 0, 4 ve 12 saat ve ayrıca ikinci aşılamadan 240 saat sonra kan örnekleri, atlardan alınmıştır. Protein şeritleri SDS-PAGE metodu ile analize edilmiştir. Test süresi boyunca, protein şeritlerindeki hareketlilik ılımlı seviyede kalmıştır. Periferal beyaz kan hücrelerindeki (PWBC) sitokinlerin mRNA ekspresyonu quantitative TaqMan®Real-Time RT-PCR (qRT-PCR) tekniği ile ölçülmüştür. Sonuçta, ateşin sebebi adjuvant değildir fakat aşı testi boyunca görülen ateş cevabı equine influenza viruslarının antijenleri ya da tetanoz toksoidi tarafından oluşturulmuş olabilir. PWBC deki sitokin ekspresyonu, aşılamadan sonra açıkca yükselmiştir. TNF-α mRNA ekspresyonu ikinci aşılamadan sonra birinci aşılamadan sonrakine göre daha güçlü indukte edilmiş görülmektedir

Anahtar Kelimeler

• Sitokinler, at, interleukin, qRT-PCR, aşı.

Kaynakça

1. Ainsworth DM, Appleton JA, Eicker SW, Luce R, Flaminio MJ, Antczak DF, 2003. The effect of c o n c e n t r a t i o n s interferon-gamma and interleukin-4 in equine pulmonary and peripheral blood mononuclear cells. Vet Immunol Immunopathol, 91: 61- 71. on mRNA o f
2. i n t e r l e u k i n - 1 2 ,
3. Bustın SA, 2000. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol, 25: 169-193.
4. Chamber TM, 1994. New approaches to control of equine influenza. Equine infectious diseases VII: Proceedings of the Seventh International Conference, June 8-11, Tokyo, Japan pp.1994-305.
5. Colahan PT, Kollias-Baker C, Leutenegger CM, Jones JH, 2002. Does training affect mRNA transciption for cytokine production in circulating leucocytes. Equine Vet J Suppl, 34: 154-158.
6. Dallman MJ, Montgomery RA, Larsen CP, Wanders A, Wells AF, 1991. Cytokine gene expression: Analysis using northern blotting, polymerase chain reaction, polymerase chain reaction and in situ hybridization. Immunol Rev, 119: 163-179.
7. European Pharmacopoeia, 1998. Monograph on Equine Influenza Vaccines, 3rd adn.
8. Gibson UE, Heid CA, Williams PM, 1996. A novel method for real time quantitative RT- PCR. Genome Res, 6: 995-1001.

9. Giguère S and Prescott JF, 1999. Quantitation of equine cytokine m RNA expression by reverse transcription-competitive polymerase chain reaction. Vet Immunol Immunopathol, 67:1-15.
10. Giguère S, Viel L, Lee E, MacKay RJ, Hernan- dez J, Franchini M, 2002. Cytokine induction in pulmonary airways of horses with heaves and effect of therapy with inhaled fluticasone propionate. Vet. Immunol Immunopathol, 85: 147-58.
11. Heid CA, Stevens J, Livak KJ, Williams PM, 1996. Real time quantitative PCR. Genome Res, 6: 986– 994.
12. Kabilan L, Andersson G, Lolli F, Ekre HP, Olsson T, Troye-Blomberg M, 1990. Detection of intracellular expression and secretion of interferon-gamma at the single-cell level after activation of human T-cells with tetanus toxoid in vitro. Europ J Immunol, 20: 1085-1089.
13. Karge WH, Schaefer EJ, Ordovas JM, 1998. Quantification of mRNA by polymerase chain reaction (PCR) using an internal standard and a nonradioactive detection method. Methods Mol Biol, 110: 43-61.
14. Kruse N, Pette M, Toyka K, Rieckmann P.1997. Quantification of cytokine mRNA expression by RT PCR in samples of previously frozen blood. J Immunol Methods, 210: 195–203
15. Laemmli UK 1970. Cleavage of structural proteins during theassembly of the head of bacteriophage T4. Nature, 227:680–685.
16. Leutenegger C M, Mislin CN, Sigrist B, Ehengruber MU, Hofmann-Lehmann R, Lutz H, 1999. Quantitative real-time PCR for the measurement of feline cytokine mRNA..Vet Immunol. Immunopathol, 71: 291–93.
17. Mackay RJ, Lester GD, 1992. Induction of the acute-phase cytokine, hepatocyte stimulating factor/interleukin-6, in the circulation of horses treated with ebdotoxin. Am J Vet Res, 53, 1285-1289.
18. Mena A, Ioannou PX, Kessel VA, Hurk SVDL- VD, Popowych Y, Babiuk AL, Godson LD, 2002 Th1/Th2 biasing effects of vaccination in cattle as determined by real-time PCR. J Immunol Methods, 263: 11 – 21.
19. Minke JM, Audonnet JC, Jessett DM, Fischer L, Guigal PM, Coupier H, Pardo MC, Taylor J, Tartaglia J, Mumford JA, 2000. Canarypox as vector for influenza and EHV-1 genes: challenges and rewards, 2nd International Veterinary Vaccines and Diagnostics Conference, Oxford, p.36.
20. Morris DD, Moore JN, 1991. Tumor necrosis factor activity in serum from neonatal foals with presumed septicaemia. J Am Vet Med Assoc, 199: 1584-1588.
21. Mumford JA, Wilson H, Hannant D, Jessett DM, 1994. Antigenicity and immunogenicity of equine influenza vaccines containing a Carbomer adjuvant. Epiemiol Infect, 112: 421-437.
22. Mosmann TR, Sad S, 1996. The expanding universe of T-cell subsets: Th1, Th2 and more. Immunol Today, 17: 138-146.
23. Newton JR, Park AW, Wood JLN, 2004. Maximizing the benefits of vaccination against Equine Influenza. In: P.Lekeux (ed) Equine Respiarory Disease. Publsher: International Veterinary Information Service (www.ivis.org), Ithaca, New York, USA.
24. Overbergh L, Valckx D, Waer M, Mathieu C, 1999. Quantification of murine cytokine mRNAs using real time quantitative reverse transcriptase PCR. Cytokine, 11, 4: 305–312.
25. Pedersen LG, Castelruiz Y, Jacobsen S, Aasted B, 2002. Identification of monoclonal antibodies that cross-react with cytokines from different animal species. Vet Immunol and Immunopathol, 88: 111–122.
26. Seethanathan P, Buttoms GD, Schafer K, 1990. Characterization of release of tumor necrosis factor, interleukin-1 and superoxide anion from equine white blood cells in response to endotoxin. Am J Vet Res, 51: 1221-1225.
27. Stordeur P, Zhoua L, Bylb B, Broheta F, Burnyc W, Grootec DD, Polld VDT, Goldmana M, 2003. Immune monitoring in whole blood using real-time PCR. J Immunol Methods, 276: 69– 77.
28. Studdert MJ. (1996): Virus infections of Equines. Studdert MJ, Ed. Virus Infections of Vertebrates. Elsevier Science, Amsterdam, pp. 9-37.
29. Tremblay GM, Ferland C, Lapointe JM, Vrins A, Lavoie JP, Cormier Y, 1993. Effect of stabling on bronchoalveolar cells obtained from normal and COPD horses. Equine Vet J, 25: 194–197.
30. Van den Hoven R, Duvigneaub JCR, Hartl T, Riedelberger K, Teinfalt M, Gemeiner M, 2004. The Expression of Messenger RNA for Tumor Necrosis Factor-α; Interleukins 1β, 6, 8, and 10; Tissue Growth Factor-β; and Interferon-γ in Bronchoalveolar Lavage Fluid Cells From Horses Challenged With and Treated With
31. Zhang Z, Carriero N, Gerstein M, 2004. Comparative pseudogenes in the mouse and human genomes. Trends Gen, 20: 62-67.