Yüksek İnfektif Titre ve 146S FMD Virus Partikülü Eldesi İçin Uygun BHK-21 Hücre Kültürü Hatlarının Saptanması
Abstract
Şap Hastalığı (FMD), çift tırnaklı hayvanların en bulaşıcı hastalıklarından biri olup duyarlı hayvanlarda ciddi ekonomik kayba neden olma konusunda büyük bir potansiyele sahiptir. FMD hastalığına karşı mücadelede inaktif FMD aşıları yaygın olarak kullanılmaktadır. Bu çalışma, FMD aşı üretiminde fazla miktarda 146S FMD (O, A ve Asia-1) virüs partikülü içeren virüs süspansiyonu elde etmek için uygun BHK-21 hücre hatlarının tespiti amacıyla yapıldı. Bu amaçla şu anda FMD aşı üretiminde kullanılan BHK-21 An30 hücre kültürüne ek olarak, BHK-21 An73 hücre hatları 20 kez pasajlandı. Her 5 pasajda bir, FMD virüsünün O, A ve Asia-1 serotiplerinin enfektif titresi ve 146S virüs partikül miktarları ölçüldü. Ayrıca, her iki hücre hattının 20. pasaj seviyelerinde karyotip analizi yapıldı ve hücrelerin morfolojik özelliklerinde ve kromozom yapılarında önemli bir değişiklik olmadığı saptandı. FMD virüsünün O, A ve Asia-1 serotiplerinin BHK-21 An73 hücre hattında elde edilen enfektif titre ve 146S virüs partikül miktarlarının, BHK-21 An30 hücre hattında saptanan değerlerden yüksek olduğu belirlendi. FMD virüsünün O, A ve Asia-1 serotiplerinin 146S virüs partikül miktarlarındaki artışların istatistiksel olarak anlamlı olduğu saptandı (p <0.05). Sonuç olarak, FMD virusuna karşı en duyarlı BHK-21 hücre hatlarının önceden belirlenmesi ile aşı üretim prosesinde yüksek titrede FMD 146S virus partikülü elde edilebilecektir. Aşı üretim kapasitesinin artmasından dolayı üreticiye ekonomik olarak önemli katkı sağlayacağı kanaatine varıldı.
Keywords
Supporting Institution
Tarımsal Araştırmalar ve politikalar Genel müdürlüğü (TAGEM)
Project Number
TAGEM/HSGYAD/16/A02/P02/78
References
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- Referans 3. S, Zhang Z, Donaldson AI, Garland AJM. 2003: The pathogenesis and diagnosis of foot and mouth disease. J Comp Path, 129, 1-36.
- Referans 4. Ali SM, Ismail AH, Soliman EM, Hanaa AM, 2013: Studies on growth kinetics of the FMDV serotype SAT-2 Egyptian strain in cell culture. J Vet Adv, 3(2), 92-97.
- Referans 5. Archettı I, Amadorı M, Donn A, Salt J, Lodettı E, 1995: Detection of foot-and-mouth disease virus-infected cattle by assessment of antibody response in oropharyngeal fluids. J Clin Microbiol, 33(1), 79-84.
- Referans 6. Baer A, Kehn-Hall K, 2014: Viral concentration determination through plaque assays. J Vis Exp, 93, 1-10.
- Referans 7. Bartelling SJ, 2002: Development and performance of inactivated vaccines against foot and mouth disease. Rev Sci Tech Off Int Epiz, 21, 577-588.
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- Referans 9. Berg K, Mohr J, 1963: Genetics of the Lp system. Acta Genet, 13(4), 349-360.
- Referans 10. Callens M, De Clercq K, 1997: Differentiation of the seven serotypes of foot-and-mouth disease virus by reverse transcriptase polymerase chain reaction. J Virol Methods, 67, 35-44.
- Referans 11. Chapman WG, Stewart DL, 1962: Growth of a cloned strain of hamster kidney cells in suspended cultures and their susceptibility to the virus of foot-and-mouth disease. Nature. 195, 1163.
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- Referans 13 Çokçalışkan C, Türkoğlu T, Sareyyüpoğlu B, Uzunlu E, Babak A., et al, 2016: Qs-21 enhances the early antibody response to oil adjuvant foot and mouth disease vaccine in cattle. Clin Exp Vac Res, 5,138-147.
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- Referans 15. Freshney RI, 2010: A manual of basic technique and specialized application. Wiley-Blackwell, A John Wiley&Sons, Inc. Pub.
- Referans 16. Grubman MJ, Baxt B, 2004: Foot and mouth disease. Clin Microbiol Rev, 17(2), 465-493.
- Referans 17. Gunetti M, Castiglia S, Rustichelli D, Mareschi K, Sanavio F et al, 2012: Validation of analytical methods in GMP: the dis¬posable fast read 102W device, an alternative practical ap¬proach for cell counting. J Trans Med, 31,10-112.
- Referans 18. Harmsen MM, Fitjen HPD, Westra DF, Coco-Martin JM, 2011: Effect of thiomersal on dissociation of intact (146s) foot-and-mouth disease virions into 12s particles as assessed by novel elisas specific for either 146s or 12s particles. Vaccine, 29, 2682-2690.
- Referans 19. Jamal SM, Belsham GJ, 2013: Foot-and-mouth disease: past, present and future. Vet Res, 44, 116.
- Referans 20. Jerome P, Howard LB, 1964: Production and purification of milligram amounts of foot-and-mouth disease virus from baby hamster kidney cell cultures. J Appl Microbiol, 12 (4), 368-373.
- Referans 21. Nawaz Z, Siddique AB, Zahoor MA, Bilal Aslam, B, Zahoor MK et al, 2019: Detectıon of foot and mouth dısease vırus sheddıng in mılk of apparently healthy buffaloes and cattle of Punjab, Pakistan. Buffalo Bull, 38(2), 255-261.
- Referans 22. OIE, 2017: OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Terrestrial Manual: foot and mouth disease: Chapter 3.1.8, (May), 433-464.
- Referans 23. Özbilge BB., Gülyaz V, Taşçene N, Yılmaz Ş, Gültekin Y et al, 2020. Determination of the effects of enrofloxacin, linco-spectinand florfenicol antibiotics on BHK-21 cell culture and FMD 146s virus particles-ınfective titers. Etlik Vet Mikrobiyol Derg, 31(1), 7-19.
- Referans 24. Paton DJ, Taylor G, 2011: Developing vaccines against foot-and-mouth disease and some other exotic viral diseases of livestock. Phil Trans R Soc B, 366(1579), 2774–2781.
- Referans 25. Rahman SU, Rabbani M, Sahidullah K, Muhammed Z, 2007: Studies on in vitro culture characteristics of adherent baby hamster kidney-21(BHK-21) cell line. Int J Agri Biol, 9(6), 821-826.
- Referans 26. Rweyemamu MM, Umehara O, Giorci W, Medeiros R, Lucca DN et al, 1989: Effect of formaldehyde and binary ethyleneimine (BEI) on the integrity of foot and mouth disease virus capsid. Rev Sci Tech Off Int Epiz, 8(3), 747-764.
- Referans 27. Sareyyüpoğlu B, Gülyaz V, Cokçalışkan C, Ünal Y, Çökülgen T, et al, 2019: Effect of fmd vaccination schedule of dams on the level and duration of maternally derived antibodies. Vet Immunol Immunopathol, Vol, 217.
- Referans 28. Shirai J, Chatchawanchonteera A, Sinsuwongwat W, Makarasen P, Sugimura T, 1990: Estimation of 146s particles in foot and mouth disease virus (FMDV) vaccine by using computer analysing system. Jpn J Vet Sci, 52(3), 621-630.
- Referans 29. Xiao Y, Chen HY, Wang Y, Yin B, Lv C et al, 2016: Large-scale production of foot-and-mouth disease virus (serotype Asia-1) VLP vaccine in escherichia coli and protection potency evaluation in cattle. BMC Biotechnology, 16: 56.
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Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles
Abstract
Foot and mouth disease (FMD) is one the most contagious diseases in cloven-hoofed animals and has a great potential for causing severe economic loss in susceptible animals. In the fight against FMD disease, inactive FMD vaccines are widely used. This study was carried out for the detection of suitable BHK-21 cell lines in order to get virus suspensions containing excess amounts of 146S FMD virus serotypes O, A and Asia-1, for FMD vaccine production. For this purpose, in addition to the BHK-21 An30 cell culture currently used in FMD vaccine production, BHK-21 An73 cell lines were passaged 20 times. In every 5 passages, infective titers and 146S virus particle quantities of FMD virus serotypes O, A and Asia-1 were measured. Additionally, karyotype analysis was performed on both cells at the 20th passage levels and the tests revealed that there was no significant change in the morphological features and chromosomal structures of the cells.
It was determined that the infective titer and 146S virus particle quantities of serotypes O, A and Asia-1 of FMD virus were very high in the BHK-21 An73 cell line than they are available in the BHK-21 An30 cell line. The increases of 146S virus particle quantities of serotypes O, A and Asia-1 of FMD virus were statistically significant (p <0.05).
As a result, high titer FMD 146S virus particles can be obtained in the vaccine production process by pre-determining the BHK-21 cell lines that are most sensitive to the FMD virus. Due to the increase in vaccine production capacity, it was concluded that it would make an important economic contribution to the producer.
Keywords
Project Number
TAGEM/HSGYAD/16/A02/P02/78
References
- Referans 1. Abbas F, Khan FA, Ahmad F, Hussain A, Ahmad M et al, 2011: Production of foot and mouth disease virus vaccine (O Type) on BHK-21. cell line, Iğdır Univ J Ins Sci Tech, 1(2), 155-159.
- Referans 2. S, Samuel AR, 2000: Identification of antigenic epitopes on the foot and mouth disease virus isolate O1/Manisa/Turkey/1969 using monoclonal antibodies. Rev Sci Tech, 19, 744-753.
- Referans 3. S, Zhang Z, Donaldson AI, Garland AJM. 2003: The pathogenesis and diagnosis of foot and mouth disease. J Comp Path, 129, 1-36.
- Referans 4. Ali SM, Ismail AH, Soliman EM, Hanaa AM, 2013: Studies on growth kinetics of the FMDV serotype SAT-2 Egyptian strain in cell culture. J Vet Adv, 3(2), 92-97.
- Referans 5. Archettı I, Amadorı M, Donn A, Salt J, Lodettı E, 1995: Detection of foot-and-mouth disease virus-infected cattle by assessment of antibody response in oropharyngeal fluids. J Clin Microbiol, 33(1), 79-84.
- Referans 6. Baer A, Kehn-Hall K, 2014: Viral concentration determination through plaque assays. J Vis Exp, 93, 1-10.
- Referans 7. Bartelling SJ, 2002: Development and performance of inactivated vaccines against foot and mouth disease. Rev Sci Tech Off Int Epiz, 21, 577-588.
- Referans 8. Bartelling SJ, Vreeswijk J, 1991: Developments in foot-and-mouth disease vaccines. Vaccine, 9(2), 5-88.
- Referans 9. Berg K, Mohr J, 1963: Genetics of the Lp system. Acta Genet, 13(4), 349-360.
- Referans 10. Callens M, De Clercq K, 1997: Differentiation of the seven serotypes of foot-and-mouth disease virus by reverse transcriptase polymerase chain reaction. J Virol Methods, 67, 35-44.
- Referans 11. Chapman WG, Stewart DL, 1962: Growth of a cloned strain of hamster kidney cells in suspended cultures and their susceptibility to the virus of foot-and-mouth disease. Nature. 195, 1163.
- Referans 12. Czelleng F, Zsitvay K, Egyhazi ZS, Baranyi M, Fazekas A, 1987: Comparative analysis of BHK-21 cell lines of virüs strains of foot-and-mounth disease. Arch Exper Vet Med, 41(6), 701-796.
- Referans 13 Çokçalışkan C, Türkoğlu T, Sareyyüpoğlu B, Uzunlu E, Babak A., et al, 2016: Qs-21 enhances the early antibody response to oil adjuvant foot and mouth disease vaccine in cattle. Clin Exp Vac Res, 5,138-147.
- Referans 14. Fransis BMJ, Fry CM, Rowlands DJ, Brown F, Bitte JL et al, 1987: Immunological priming with synthetic peptides of foot-and-mouth disease. Immunology, 61(1), 1-6.
- Referans 15. Freshney RI, 2010: A manual of basic technique and specialized application. Wiley-Blackwell, A John Wiley&Sons, Inc. Pub.
- Referans 16. Grubman MJ, Baxt B, 2004: Foot and mouth disease. Clin Microbiol Rev, 17(2), 465-493.
- Referans 17. Gunetti M, Castiglia S, Rustichelli D, Mareschi K, Sanavio F et al, 2012: Validation of analytical methods in GMP: the dis¬posable fast read 102W device, an alternative practical ap¬proach for cell counting. J Trans Med, 31,10-112.
- Referans 18. Harmsen MM, Fitjen HPD, Westra DF, Coco-Martin JM, 2011: Effect of thiomersal on dissociation of intact (146s) foot-and-mouth disease virions into 12s particles as assessed by novel elisas specific for either 146s or 12s particles. Vaccine, 29, 2682-2690.
- Referans 19. Jamal SM, Belsham GJ, 2013: Foot-and-mouth disease: past, present and future. Vet Res, 44, 116.
- Referans 20. Jerome P, Howard LB, 1964: Production and purification of milligram amounts of foot-and-mouth disease virus from baby hamster kidney cell cultures. J Appl Microbiol, 12 (4), 368-373.
- Referans 21. Nawaz Z, Siddique AB, Zahoor MA, Bilal Aslam, B, Zahoor MK et al, 2019: Detectıon of foot and mouth dısease vırus sheddıng in mılk of apparently healthy buffaloes and cattle of Punjab, Pakistan. Buffalo Bull, 38(2), 255-261.
- Referans 22. OIE, 2017: OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Terrestrial Manual: foot and mouth disease: Chapter 3.1.8, (May), 433-464.
- Referans 23. Özbilge BB., Gülyaz V, Taşçene N, Yılmaz Ş, Gültekin Y et al, 2020. Determination of the effects of enrofloxacin, linco-spectinand florfenicol antibiotics on BHK-21 cell culture and FMD 146s virus particles-ınfective titers. Etlik Vet Mikrobiyol Derg, 31(1), 7-19.
- Referans 24. Paton DJ, Taylor G, 2011: Developing vaccines against foot-and-mouth disease and some other exotic viral diseases of livestock. Phil Trans R Soc B, 366(1579), 2774–2781.
- Referans 25. Rahman SU, Rabbani M, Sahidullah K, Muhammed Z, 2007: Studies on in vitro culture characteristics of adherent baby hamster kidney-21(BHK-21) cell line. Int J Agri Biol, 9(6), 821-826.
- Referans 26. Rweyemamu MM, Umehara O, Giorci W, Medeiros R, Lucca DN et al, 1989: Effect of formaldehyde and binary ethyleneimine (BEI) on the integrity of foot and mouth disease virus capsid. Rev Sci Tech Off Int Epiz, 8(3), 747-764.
- Referans 27. Sareyyüpoğlu B, Gülyaz V, Cokçalışkan C, Ünal Y, Çökülgen T, et al, 2019: Effect of fmd vaccination schedule of dams on the level and duration of maternally derived antibodies. Vet Immunol Immunopathol, Vol, 217.
- Referans 28. Shirai J, Chatchawanchonteera A, Sinsuwongwat W, Makarasen P, Sugimura T, 1990: Estimation of 146s particles in foot and mouth disease virus (FMDV) vaccine by using computer analysing system. Jpn J Vet Sci, 52(3), 621-630.
- Referans 29. Xiao Y, Chen HY, Wang Y, Yin B, Lv C et al, 2016: Large-scale production of foot-and-mouth disease virus (serotype Asia-1) VLP vaccine in escherichia coli and protection potency evaluation in cattle. BMC Biotechnology, 16: 56.
- Referans 30. Van Wezel AL, 1967: Growth of cell-strains and primary cells on microcarriers in homogeneous culture. Nature, 216; 64.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Journal Section | Research |
| Authors | |
| Project Number | TAGEM/HSGYAD/16/A02/P02/78 |
| Publication Date | June 22, 2021 |
| Submission Date | October 13, 2020 |
| Acceptance Date | February 23, 2021 |
| Published in Issue | Year 2021 Volume: 10 Issue: 1 |
