为了研究雏鸡注射不同浓度刺五加多糖(ASPS)后脾脏中T、B淋巴细胞定位分布的变化情况,将60只1日龄雏鸡随机分为3组:对照组、低剂量组和高剂量组,每组20只,10日龄起对低、高剂量组雏鸡分别皮下注射免疫50和200 mg/m L的ASPS 0.2 m L,对照组雏鸡以相同方式注射等体积的生理盐水,连续注射4 d。于末次免疫后第7天和第14天,从每组中分别随机选取5只雏鸡,取其脾脏制作冰冻切片,采用免疫组织化学方法检测CD3+T淋巴细胞和Bu-1+B淋巴细胞的定位分布。结果显示,与对照组比较,低、高剂量组在注射后的7 d和14 d ASPS均能增加脾脏中CD3+T淋巴细胞和Bu-1+B淋巴细胞的数量,由CD3+T淋巴细胞构成的脾脏特征性结构-动脉周围淋巴鞘(PALS)的面积也显著增加,红髓中CD3+T淋巴细胞也明显增多,分布范围逐渐增加;同时,由Bu-1+B淋巴细胞构成的椭球周围淋巴鞘(EALS)的面积也明显增加,红髓和白髓中的浆细胞数量显著增多,并在注射后14 d时出现生发中心。与对照组比较,高剂量组和注射后14 d时上述变化趋势更为明显。结果表明,ASPS能够增加脾脏中T、B淋巴细胞的数量,并且能够影响其在PALS、EALS等脾脏特征性结构中的定位分布,从而从组织学角度进一步证明ASPS对鸡免疫功能具有显著的增强作用。
The aim of this study was to investigate the effect of different concentration ASPS on distribution of T and B lymphocytes in chicken spleen. The chicks were divided into control group,low- dose group and high- dose group. There were 20 chicks in each group. The chicks in low- dose group and high- dose group respectively subcutaneous injected 0. 2 m L ASPS with the dose of 50 and 200 mg / m L,The control group were administered saline and treated as the same way as the fore groups. The same operation was repeated 4 d. Five chicken spleen were removed from each group on the day 4 and 14 after the last day of immunization. The localization of T and B lymphocytes in chicken spleen were detected by Immunohistochemistry staining. The result showed the number of CD3+T lymphocytes and Bu- 1+B lymphocytes in the spleen were increased in both low dose group and high dose group compared with the control group after injection for 7 d and 14 d. The area of periarterial lymphatic sheath which was composed of CD3+T lymphocytes was also increased,meanwhile CD3+T lymphocytes in the red pulp also increased obviously,and the distribution range gradually increased. At the same time,the area of periellipsoid lymphatic sheath which is composed of Bu- 1+B lymphocytes was also increased,and the number of plasma cells in red pulp and white pulp was significantly increased,and even the germinal center were appeared in 14 d after injection. Compared with the control group,the change tendency of high dose group and 14 d after injection were more obvious. In conclusion,ASPS can increase the number of T and B lymphocytes in spleen,and can affect their localization in the characteristic structures of spleen like periarterial lymphatic sheath and periellipsoid lymphatic sheath,and further prove that ASPS have significant enhancement effect on immune function of chicken from the histological angle.