摘要
无菌采取犬抗凝血,分离淋巴细胞并制备成不同浓度。应用正交试验,探讨不同浓度淋巴细胞、植物血凝素P(PHA-P)、犊牛血清(FCS)对淋巴细胞增殖的影响。在此基础上,研究不同浓度犬、猪转移因子(transferfactor,TF)对犬淋巴细胞增殖的影响,增殖的结果用MTT法测定。结果表明,当犬淋巴细胞浓度为5×106个/ml,PHA-P为12.5μg/ml,FCS为10%时,淋巴细胞增殖效果最佳。在最佳培养条件下,犬、猪TF浓度在0.195￣25mg/ml时,对淋巴细胞转化增殖均有促进作用,但犬、猪TF单独或与PHA-P共同作用时对淋巴细胞增殖的最佳刺激浓度均为1.56mg/ml,且犬、猪TF单独作用于淋巴细胞的效应明显优于与PHA-P的协同作用。结果证实,同源或异源TF对犬淋巴细胞增殖有良好的刺激效果,同时研究结果为异源TF在临床上应用于犬病毒性疾病的防治提供了实验依据。
Pick-off blood with anticoagulant aseptically and the lymphocytes were isolated. Effects of FCS,PBMC,PHA-P of different concentration on promote lymphocytes proliferation were tested by orthogonal test. On the base of the best level combination, different concentration canine or pig TF was investigated to detect its effect on the activation of lymphocytes by MTT essay. The result of experiment demonstrate that the best levels combination of canine lymphocyte, PHA-P, calf blood serum were 2.5×106 unit/ml,6.25μg/ml,10% respectively. Under this optimum cultural condition, the lymphocytes remarkable proliferated between 0.195~25mg/ml of canine and pig TF,but the best stimulate concentration of the canine and pig TF only or it with PHA-P is 1.56mg/ml. Furthermore, it is confirmed that lymphocytes developed by the canine and pig TF single was much better than they cooperated with PHA-P. The results confirm that the same or difficult race TF's effect was obvious and provides a basis for further application on the difficult race TF preventing and curing canine's virus disease.
出处
《中国农学通报》
CSCD
2005年第6期26-28,62,共4页
Chinese Agricultural Science Bulletin
基金
陕西省农业分子生物学重点实验室项目(2002.09)
公安部科技局科研项目(2004.06)资助
