Institute of Traditional Chinese Veterinary Medicine, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China.
Int J Biol Macromol. 2013 Aug;59:178-83. doi: 10.1016/j.ijbiomac.2013.04.007. Epub 2013 Apr 12.
Cordyceps militaris polysaccharide (CMP) was prepared by water decoction and ethanol precipitation. The fractional CMP40 and CMP50 were extracted from the CMP solution by stepwise precipitation with ethanol at 40% and 50% of working concentration, respectively. The immune-enhancing activities of two polysaccharides were researched. In vitro experiment, the effects of two polysaccharides on chicken peripheral lymphocyte proliferation were determined by MTT assay. The result displayed that two polysaccharides could significantly stimulate lymphocyte proliferation, the action of CMP40 was significantly or numerically stronger than those of CMP50. In vivo experiment, 320 14-day-old chickens were averagely divided into eight groups. The chickens, except blank control (BC) group, were vaccinated with Newcastle disease vaccine, repeated vaccination at 28 days old. At the same time of the first vaccination, the chickens in three CMP40 fraction groups and three CMP50 fraction groups were injected respectively with the polysaccharide at low, medium and high concentrations, in vaccination control (VC) and BC group, with equal volume of physiological saline, once a day for three successive days. On days 7, 14, 21, 28, 35 and 42 after the first vaccination, the lymphocyte proliferation, serum antibody titre and interferon-gamma and interleukin-4 were measured. The results showed that CMP40 and CMP50 at suitable dose could significantly promote lymphocyte proliferation, enhance serum antibody titre, and improve serum interferon-gamma and interleukin-4 concentrations. It indicated that CMP40 fraction and CMP50 fraction could significantly improve the immune efficacy of Newcastle disease vaccine, and would be as the candidate of a new-type immune adjuvant.
蛹虫草多糖（CMP）采用水提醇沉法制备。将 CMP 溶液分别用 40%和 50%工作浓度的乙醇分步沉淀，提取出分级 CMP40 和 CMP50。研究了两种多糖的免疫增强活性。体外实验，采用 MTT 法测定两种多糖对鸡外周血淋巴细胞增殖的影响。结果表明，两种多糖均能显著刺激淋巴细胞增殖，CMP40 的作用明显或数值上强于 CMP50。体内实验，将 320 只 14 日龄鸡平均分为 8 组。除空白对照组（BC）外，其余鸡均接种新城疫疫苗，28 日龄重复接种。首次免疫时，三个 CMP40 级分组和三个 CMP50 级分组的鸡分别注射低、中、高浓度的多糖，在免疫对照组（VC）和 BC 组注射等量生理盐水，连续 3 天，每天 1 次。首次免疫后第 7、14、21、28、35 和 42 天，测定淋巴细胞增殖、血清抗体效价以及干扰素-γ和白细胞介素-4。结果表明，适宜剂量的 CMP40 和 CMP50 可显著促进淋巴细胞增殖，提高血清抗体效价，改善血清干扰素-γ和白细胞介素-4浓度。这表明 CMP40 级分和 CMP50 级分可显著提高新城疫疫苗的免疫效果，有望成为新型免疫佐剂的候选物。
