عنوان مقاله [English]
There are many guidelines for hemolysin production, but for production with higher titers, standardization of procedures is essential. In this study, for the first time in the country, the hemolysin was produced by whole sheep blood intradermal injection to laboratory rabbits and was compared with the intravenous injection of 2% washed sheep red blood cells routine method. The produced hemolysins were titerd by kolmer method. In whole blood intradermal injection method, the rabbit anti-sheep hemolysin titer was at least 1:9600 whereas in 2% washed cells intravenous injection method, obtained 1:1200. The obtained results of the three replicates were taken on two methods listed were exactly the same as before. In order to increasing the products stability, they was lyophilized and were kept in -20oC. For survey in maintained products stability time, once every six months for 2 years, both the products, were tittered. To evaluate the in vitro activity, they were used in serological tests. In this survey, the hemolysin produced by whole sheep blood intradermal injection, in all cases, and even after 2 years of production, there is no reduction in titer and it was very favorable in serologic tests, but the product was prepared by intravenous injection, showed sharp decline in titer and it was not an acceptable result in serologic tests. According to the results, the whole blood intradermal method occording to this protocol can be replaced to routingly 2% washed cells intravenous injection for production of anti-sheep hemolysin with high titer, stability and effect.
1-Archetti, I., Tittarelli, C., Cerioli, M., Brivio, R., Grilli, G. and Lavazza, A.(2008) Serum chemistry and hematology values in commercial rabbits: preliminary data from industrial farms in northern Italy, 9th World Rabbit Congress, Verona, Italy.
2-Berglund, K. (1956) Hemolysin response in the rat after a single intraperitoneal injection of sheep erythrocytes. Acta Pathology and Microbiology of Scandinavy 38(3): 220-230.
3-Bratcher, R.L. (1979) High responder rabbits to SRBC, A familiar incidence, The Journal of Immunology, 122 (1) 49-53.
4-Darter, L.A. (1953) Procedure for production of anti-sheep hemolysin. Journal of Laboratory Clinical Medicine 41: 653-654.
5- Ethics of research involving animals, (2005) Published by Nuffield Council on Bioethics, 28 Bedford Square London, WC1B 3JS.
6-Fish, R.R.E., Brown, M.J., Danneman, P.J. and Karas, A.Z. (2008) Anesthesia and Analgesia in Laboratory Animals, Second Edition , American College of Laboratory Animal Medicine.
7-Harkness, J.E., Turner,P.V.and Woude,S,V. (2010) Harkness and Wagner's Biology and Medicine of Rabbits and Rodents, 5th revised edition, Iowa State University Press.
8-Henry, J.B. (2001) Clinical diagnosis and management by laboratory methods, 20th ed. USA Press, 908.
9-Kohn, H.I. (1951) Effect of x rays upon hemolysin production in the rat. Journal of Immunology 66(5): 525-533.
10-Koyama, J. and Nashimoto, H. (1959) Immunological studies on rabbit hemolysin to sheep red cells. I. The electrophoretic separation of rabbit antisera to sheep red cells. Japanese Journal of Experimental Medicine 29: 551-559.
11- Koyama, J. and Nashimoto, H. (1959) Immunological studies on rabbit hemolysin to sheep red cells. II. The separation and purification of the hemolysin with an anion exchanger: diethylaminoethyl cellulose. Japanese Journal of Experimental Medicine 29: 561-570.
12- Manning, P.J., Ringler, D.H. and Newcomer, C. E. (1994) The Biology of the Laboratory Rabbit, 2nd edition, Academic Press, Orlando, Florida.
13- Mark A. Suckow,M.A., Schroeder, V. and. Douglas, F.A. (2010) The Laboratory Rabbit, Second Edition, CRC Press.
14-Mukani, W.O., Nyangao, J.N.M., Kiamani, J.K., Omuse, J.K. (1995) Studies on the haemolytic complement of the dromedary camel (Camelus dromedaries), I. Classical pathway haemolytic activity in serum, Veterinary Immunilogy and Immunopathology, 46: 339.
15-Nakamura, R. M., Burek, C.L., Cook, L., Folds, J.D. (1998) Clinical diagnostic immunology: Protocols in quality assurance and standardization, USA Press, 94-95.
16-Oyama, J., Nashimoto, Y. and Masuyoshi, T. (1961) Immunological studies on the anti-sheep-erythrocyte hemolysin of the rabbit. Nippon Saikingaku Zasshi 16: 641-642.
17-Palmer, D.F. (1980) Complement fixation test, Manual of clinical immunology (2nd edition), 35-47.
18-Pike, R.M. and Schulze, M. L. (1960) Agglutination in rheumatoid arthritis serum of sheep cells sensitized with hemolysin and infectious mononucleosis agglutinins. Journal of Immunology 85: 523-529.
19-Rowan, A.N. (1995) Animals in research,Cardiovascular Research, 29(4):583.
20-Smith, F. and Ruth, H.J. (1955) Hemolysin production in irradiated mice given spleen or bone-marrow homogenate. Proceeding of Society in Experimental Biological Medicine 90(1): 187-191.
21- Suckow,M.A., Stevens, K.A.and Wilson, R.P. (2012) The Laboratory Rabbit, Guinea Pig, Hamster, and Other Rodents, American College of Laboratory Animal Medicine, Academic Press.
22-Taliaferro, W.H. and Taliaferro, L.G. (1950) The dynamics of hemolysin formation in intact and splenectomized rabbits. Journal of Infectious Diseases 87: 37-62.
23-Taliaferro, W.H. and Talmage, D.W. (1956) Antibodies in the rabbit with different rates of metabolic decay. Journal of Infectious Diseases 99: 21-33.
24-Talmage, D.W., Freter, G.G. and Taliaferro, W.H. (1956). The effect of whole body x radiation on the natural sheep cell hemolysin of the rabbit. Journal of Infectious Diseases 99(3): 241-245.
25 - UFAW Handbook on the Care and Management of Laboratory and Other Research Animals, (2010) 8th edition,Wiley-Blackwell.