생명과학회지 (Journal of Life Science)

  • 제20권2호
  • /
  • Pages.153-160
  • /
  • 2010
  • /
  • 1225-9918(pISSN)
  • /
  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
DOI QR코드

DOI QR Code

면역조직 화학법 및 단백질체 변화 분석을 통한 한우에서 발생한 브루셀라증의 특성

Characterization of Bovine Brucellosis in Korean Native Cattle by Means of Immunohistochemistry and Proteomics

  • Jang, Seong-Jun (College of Veterinary Medicine, Kyungpook National University) ;
  • Do, Sun-Hee (College of Veterinary Medicine, Konkuk University) ;
  • Ki, Mi-Ran (College of Veterinary Medicine, Kyungpook National University) ;
  • Hong, Il-Hwa (College of Veterinary Medicine, Kyungpook National University) ;
  • Park, Jin-Kyu (College of Veterinary Medicine, Kyungpook National University) ;
  • Cho, Yu-Jeong (College of Veterinary Medicine, Kyungpook National University) ;
  • Park, Sang-Joon (College of Veterinary Medicine, Kyungpook National University) ;
  • Kim, Tae-Hwan (College of Veterinary Medicine, Kyungpook National University) ;
  • Kwak, Dong-Mi (College of Veterinary Medicine, Kyungpook National University) ;
  • Jeong, Kyu-Shik (College of Veterinary Medicine, Kyungpook National University)
  • 투고 : 2009.09.25
  • 심사 : 2009.01.25
  • 발행 : 2010.02.28
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

면역조직 화학법 및 단백질체 변화 분석을 통한 한우에서 발생한 브루셀라증의 특성 본 연구는 브루셀라증 감염 소 혈청으로부터 분리한 항 브루셀라 면역글로블린 항체를 이용하여 조직 면역 염색을 통한 브루셀라증 진단의 활용 가능성을 조사하고 병의 발생과 관련한 기능적 진단 마커를 개발하고자 하였다. Rose-Bengal test에 대해 양성 반응을 나타내어 브루셀라증으로 진단된 17개의 케이스와 음성 반응을 나타낸 19개의 대조군 케이스에 대해 조사를 실시하였다. 본 실험실에서 분리한 항 브루셀라 항체를 이용한 면역조직화학적 반응에서 간의 중심 소엽에 위치한 간세포의 세포질, 신장의 사구체 및 관 상피에서 강한 양성 반응을 나타내었다. 감염된 소의 간과 비감염 대조군의 간의 2차원 전기 영동법에 의한 단백질체를 비교 분석한 결과, 발현량이 대조군에 비해 유의적으로 증가한 5개의 단백질 스팟과 반대로 대조군에 비해 발현량이 현저히 감소한 5개의 단백질 스팟을 선별 하였다. 이 중 카탈라아제와 3-hydroxyacyl-CoA dehydrogenase의 발현 증가는 브루셀라증에 의한 장독 쇼크에 의한 산화적 스트레스 증가에 대한 방어적 반응으로 사료 되었다. 결론적으로, 항 브루셀라 면역글로불린 항체는 감염 조직의 감별 진단을 위한 좋은 진단 재료 임과, 더 나아가 단백질체학 분석을 통해 브루셀라증 진단 및 병리 연구를 위한 새로운 마커 단백질을 제시하였다.

This study was conducted to examine the utilization of immunohistochemistry using the bovine anti-brucella immunoglobulin G (IgG) antibody in the diagnosis of brucellosis and to develop a functional biomarker relation for the progress of the disease. Anti-brucella IgG antibody was purified from the affected bovine serum using an affinity chromatography. We performed our investigation on 17 cases of brucellosis and 19 control cases with negative Rose-Bengal test results. Our purified anti-brucella IgG antibody showed a positive immunoreactivity in cytoplasmic hepatocytes of the centrilobular region, and glomeruli and tubular epithelium of the kidney. The protein pattern of the affected liver versus control was analyzed by two-dimensional electrophoresis, showing a different expression pattern of proteins between the two. Five protein spots were up-regulated and another were five down-regulated in the brucellosis liver. Significant upregulaton of catalase and 3-hydroxyacyl-CoA dehydrogenase might be due to a compensatory reaction in response to the endotoxic shock of brucella. In conclusion, the anti-brucella IgG antibody may be a good tool for discriminative diagnosis of the affected tissues and proteomics data suggest new target proteins underlying a possible pathogenic mechanism of brucellosis.

키워드

참고문헌

  1. Adams, L. G. 2002. The pathology of brucellosis reflects the outcome of the battle between the host genome and the Brucella genome. Vet. Microbiol. 90, 553-561. https://doi.org/10.1016/S0378-1135(02)00235-3
  2. AIMS. 2007. Cases of Infectious Livestock Disease. Animal Infectious disease data Management System. (http:// aims.nvrqs.go.kr).
  3. Anderson, T. D., V. P. Meador, and N. F. Cheville. 1986. Pathogenesis of placentitis in the goat inoculated with Brucella abortus. I. Gross and histologic lesions. Vet. Pathol. 23, 219-226.
  4. Cantoni, G. L. and P. J. Vignos Jr. 1954. Enzyme mechanism of creatine synthesis. J. Biol. Chem. 209, 647-659.
  5. Chance, B., H. Sies, and A. Boveris. 1979. Hydroperoxide metabolism in mammalian organs. Physiol. Rev. 59, 527-605.
  6. Enright, F. M., J. V. Walker, G. Jeffers, and B. L. Deyoe. 1984. Cellular and humoral responses of Brucella abortus-infected bovine fetuses. Am. J. Vet. Res. 45, 424-430.
  7. Foster, G., A. P. MacMillan, J. Godfroid, F. Howie, S. Brew, and A. P. Patterson. 2002. A review of Brucella sp. infection of sea mammals with particular emphasis on isolates from Scotland. Vet. Microbiol. 90, 563-580. https://doi.org/10.1016/S0378-1135(02)00236-5
  8. Geetha-Habib, M., R. Noiva, H. A. Kaplan, and W. J. Lennarz. 1988. Glycosylation site binding protein, a component of oligosaccharyl transferase, is highly similar to three other 57 kd luminal proteins of the ER. Cell 54, 1053-1060. https://doi.org/10.1016/0092-8674(88)90120-1
  9. Gingras, A. C., B. Raught, and N. Sonenberg. 1999. eIF4 initiation factors: effectors of mRNA recruitment to ribosomes and regulators of translation. Annu. Rev. Biochem. 68, 913-963. https://doi.org/10.1146/annurev.biochem.68.1.913
  10. Gorham, S. L., F. M. Enright, T. G. Snider, and E. D. Roberts. 1986. Morphologic lesions in Brucella abortus infected ovine fetuses. Vet. Pathol. 23, 331-332.
  11. Gorvel, J. P. and E. Moreno. 2002. Brucella intracelluar life from invasion to intracellular replication. Vet. Microbiol. 90, 281-297. https://doi.org/10.1016/S0378-1135(02)00214-6
  12. Hashimoto, F. and H. Hayashi. 1987. Significance of catalase in peroxisomal fatty acyl-CoA beta-oxidation. Biochim. Biophys. Acta. 921, 142-150. https://doi.org/10.1016/0005-2760(87)90180-9
  13. He, X. Y., H. Schulz, and S. Y. Yang. 1998. Ahuman brain L-3-hydroyacyl-coenzyme A dehydrogenase is identical to anamyloid beta-peptide-binding protein involved in Alzeheimer's disease. J. Biol. Chem. 273, 10741-10746. https://doi.org/10.1074/jbc.273.17.10741
  14. Hershko, A. and A. Ciechanover. 1998. The ubiquitin system. Annu. Rev. Biochem. 67, 425-479. https://doi.org/10.1146/annurev.biochem.67.1.425
  15. Jones, A. L., M. D. Hulett, and C. R. Parish. 2005. Histidinerich glycoprotein: A novel adaptor protein in plasma that modulates the immune, vascular and coagulation systems. Immunol. Cell Biol. 83, 106-118. https://doi.org/10.1111/j.1440-1711.2005.01320.x
  16. KCDC. 2006. 52 week Communicable Disease Weekly Report (CDWR). Korea Center for Disease Control & Prevention. December 24-30. 5, (52). (http://www.cdc.go.kr, http://dis. cdc.go.kr)
  17. Langen, H., D. Roder, J. F. Juranville, and M. Fountoulakis. 1997. Effect of protein application mode and acrylamide concentration on the resolution of protein spots separated by two-dimensional gel electrophoresis. Electrophoresis 18, 2085-2090. https://doi.org/10.1002/elps.1150181135
  18. Maratea, J., D. R. Ewalt, S. Jr. Frasca, J. L. Dunn, S. De Guise, L. Szkudlarek, D. J. St Aubin, and R. A. French. 2003. Evidence of Brucella sp. infection in marine mammals stranded along the coast of southern New England. J. Zoo Wildl. Med. 34, 256-61. https://doi.org/10.1638/02-053
  19. Meador, V. P., B. L. Deyoe, and N. F. Cheville. 1989. Effect of nursing on Brucella abortus infection of mammary glands of goats. Vet. Pathol. 26, 369-375.
  20. Meador, V. P., B. L. Deyoe, and N. F. Cheville. 1989. Pathogenesis of Brucella abortus infection of the mammary gland and supramammary lymph node of the goat. Vet. Pathol. 26, 357-368.
  21. Meador, V. P., D. P. Warner, and B. L. Deyoe. 1988. Distribution of Brucella abortus organisms in calves after conjunctival exposure. Am. J. Vet. Res. 49, 2015-2017.
  22. Miller, I., M. Gemeiner, B. Gesslbauer, A. Kungl, C. Piskernik, S. Haindl, S. Nurnberger, S. Bahrami, H. Redl, and A. V. Kozlov. 2006. Proteome analysis of rat liver mitochondria reveals a possible compensatory response to endotoxic shock. FEBS Lett. 580, 1257-1262. https://doi.org/10.1016/j.febslet.2006.01.040
  23. Nicoletti, P. 2002. A short history of brucellosis. Vet. Microbiol. 90, 5-9. https://doi.org/10.1016/S0378-1135(02)00209-2
  24. Ogawa, H., Y. Ishiguro, and M. Fujioka. 1983. Guanidinoacetate methyltransferase from rat liver: purification, properties, and evidence for the involvement of sulfhydryl groups for activity. Arch. Biochem. Biophys. 226, 265-275. https://doi.org/10.1016/0003-9861(83)90293-X
  25. Roubelakis, M. G., K. I. Pappa, V. Bitsika, D. Zagoura, A. Vlahou, H. A. Papadaki, A. Antsaklis, and N. P. Anagnou. 2007. Molecular and proteomic characterization of human mesenchymal stem cells derived from amniotic fluid: comparison to bone marrow mesenchymal stem cells. Stem Cells Dev. 16, 931-952. https://doi.org/10.1089/scd.2007.0036
  26. Stockler, S., B. Marescau, P. P. De Deyn, J. M. Trijbels, and F. Hanefeld. 1997. Guanidino compounds in guanidinoacetate methyltransferase deficiency, a new inborn error of creatine synthesis. Metabolism 46, 1189-1193. https://doi.org/10.1016/S0026-0495(97)90215-8
  27. Tachibana, M., K. S. Watanabe, S. Kim, Y. Omata, K. Murata, T. Hammond, and M. Watarai. 2006. Antibodies to Brucella spp. in Pacific bottlenose dolphins from the Solomon Islands. J. Wildl. Dis. 42, 412-414. https://doi.org/10.7589/0090-3558-42.2.412
  28. Tagupsky, P. and E. J. Baron. 2005. Laboratory exposure to brucellae and implications for bioterrorism. Emerging infectious diseases. Centers for disease control and prevention. 11,1180-1185. (http://www.cdc.gov/ncidod/eid/vol11no08/ 04-1197.htm) https://doi.org/10.3201/eid1108.041197
  29. Tien, I., D. C. De Vivo, D. E. Hale, J. T. R. Clarke, H. Zinman, R. Laxer, A. Shore, and S. DiMauro. 1991. Shortchain l-3-hydroxyacyl-CoA dehydrogenase deficiency in muscle: a new cause for recurrent myoglobinuria and encephalopathy. Ann. Neurol. 30, 415-419. https://doi.org/10.1002/ana.410300315
  30. Timoney, J. F., J. F. Gillespie, F. W. Scott, and J. E. Barlough. 1988. Hagan and Bruner's Microbiology and Infectious Diseases of Domestic Animals. 8th (eds.), pp. 135-152.
  31. Wakil, S. J., D. E. Green, S. Mii, and H. R. Mahler. 1954. Studies on the fatty acids oxidizing system of animal tissues. VI. beta-hydroxyacyl coenzyme A dehydrogenase. J. Biol. Chem. 207, 631-638.
  32. Wetterau, J. R., K. A. Combs, L. R. McLean, S. N. Spinner, and L. P. Aggerbeck. 1991. Protein disulfide isomerase appears necessary to maintain the catalytically active structure of the microsomal triglyceride transfer protein. Biochemistry 30, 9728-9735. https://doi.org/10.1021/bi00104a023