Malays. Appl. Biol. (2005) 34 (1): 1-7 EFFECTS OF CHITOSAN ON LYMPHOPROLIFERATION SEH, C.C., NADARAJAH, K.*, AHMAD, I.E. and ZAINAL-ABIDIN, A.H. School of Bioscience and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia 43600 Bangi ABSTRACT This study was carried out to determine the effect of fungal chitosan on the proliferation of murine lymphocytes in comparison with crustacean chitosan. Fungal chitosan (FC) was extracted from the mycelia of Absidia butleri DR whereas the crustacean chitosan (CC) is of shrimp origin. Intraperitoneal administrations of FC at 50 mg/kg or 250 mg/kg body weight for 1 week or 3 weeks showed no significant effects on the proliferative responses of lymphocytes to Con A or LPS stimulation. In contrast, administration of CC at 50 mg/kg and 250 mg/kg for 1 week and 50 mg/ kg for 3 weeks significantly enhanced proliferation of lymphocytes stimulated with LPS. Neither FC nor CC was found to affect proliferation of lymphocytes in vitro. The proliferative response was suppressed when 100 ug/ml FC was co incubated with Con A but not with LPS. Coincubation of Con A with CC at 50 ug/ml and 100 |ig/ml resulted in significant suppression of lymphoproliferation. CC at 100 jig/ml was also found to exhibit significant suppression in the proliferative response to LPS. ABSTRAK Kajian ini dilakukan untuk menentukan kesan kitosan kulat ke atas proliferasi limfosit mencit dalam perbandingan dengan kitosan krustasia. Kitosan kulat (FC) diekstrak daripada dinding sel Absidia butleri DR manakala asal kitosan krustasia (CC) dari udang. Kitosan kulat pada kepekatan 50 mg/kg atau 250 mg/kg berat badan didapati tidak memberikan sebarang kesan ke atas proliferasi limfosit aruhan Kon A atau LPS pada mencit yang diberi perlakuan selama 1 minggu ataupun 3 minggu berbanding mencit kawalan. Sebaliknya, mencit yang diperlakukan CC pada kepekatan 50 mg/kg dan 250 mg/kg berat badan selama seminggu mempamerkan peningkatan signifikan dalam respons proliferasi terhadap LPS. Akan tetapi, cuma perlakuan 50 mg/kg CC selama 3 minggu sahaja didapati memberikan peningkatan signifikan ke atas proliferasi sel limpa terhadap mitogen yang sama. Hasil kajian turul menunjukkan bahawa FC dan CC tidak mengaruh proliferasi sel limpa in vitro. Apabila Kon A dieram bersama FC pada kepekatan 100 ug/ml, penekanan proliferasi limfosit berlaku secara signifikan tetapi proliferasi sel aruhan LPS tidak dipengaruhi. Kepekatan CC yang berbeza tidak memberi kesan ke atas proliferasi sel limpa apabila dierarr bersama dengan Kon A kecuali pada kepekatan 50 ug/ml dan 100 ug/ml di mana penekanan proliferasi sel berlaku Kepekatan 100 ug/ml turut menekan proliferasi sel aruhan LPS dengan signifikan. Key words: Chitosan, lymphocyte proliferation, MTT assay, immune toxicity REFERENCES Ben-Josef, A.M., Cutright, J.L., Manavathu, E.K. & Sobel, J.D. 2003. CAN-296-P is effective against cutaneous candidiasis in guinea pigs. International Journal of Antimicrobial Agents 22: 168-171. Chung, L.Y., Schmidt, R.J., Hamlyn, P.P., Sagar, B.F., Andrews, A.M. & Turner, T.D. 1994. Biocompatibility ofpotential wound management products: fungal mycelia as a source of chitin/chitosan and their effect on the proliferation of human F1000 fibroblasts in culture. Journal of Biomedical Materials Research 28: 463-469. Koide, S.S. 1998. Chitin-chitosan: Properties, benefits and risks. Nutrition Research, 18: 1091-1101. Luster, M.I., Dean, J.H. & Moore, J.A. 1982. Evaluation of immune functions in toxicology. In: Principles and methods of toxicology. Hayes, A.W. (Ed). Raven Press, New York. Mossman, T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65: 55-63 Muzzarelli, R.A.A. & Rochetti, R. 1985. Determination of the degree of acetylation of chitosans by first derivative ultraviolet spectrophotometry. Carbohydrate Polymers 5: 461-472. Shibata, Y., Foster, L.A., Metzger, W.J. & Myrvik, Q.N. 1997. Alveolar macrophage priming by intravenous administration of chitin particles, polymers of N-acetyl-D-glucosamine, in mice. Infection and Immunity 65: 1734-1741. Su, C.H., Sun, C.S., Juan, S.W., Ho, H.O., Hu,C.H. & Sheu, M.T. 1999. Development of fungal mycelia as skin substitudes: effects on wound healing and fibroblast. Biomaterials, 20: 61-68. White, S.A., Farina, P.R. & Fulton, I. 1979. Production and isolation of chitosan from Mucor rouxii. Applied Environmental Microbiology, 38: 323-328.
