不同日龄仔猪外周血T淋巴细胞的检测与分析

摘要

To establish healthy piglets of peripheral blood lymphocyte subsets in clinical reference range, and analyze the level of immune system in different growth stages of piglets, the blood samples were collected on the 1st day, 7th day, 21st day and 35th day after weaning, respectively, and the mononuclear cells were isolated from the peripheral blood.Three color flow cytometry was used to detect the proportion of primary kinds of T lymphocyte subsets in peripheral blood mononuclear cells and in CD3+ T lymphocyte subsets, then the ratio of CD4+ /CD8+ was calculated.The proportion of CD3+ T lymphocyte subsets for peripheral blood mononuclear cells in male and female piglets were gradually increased from (38.00±10.94)% to (55.43±12.30)% and from (38.17±15.39)% to (51.57±10.60)% , respectively with the increase of age.On the 1st day and 7th day after weaning, the CD3+ T lymphocyte ratio had no significant difference between male and female piglets (P>0.05).On the 21st day after weaning, the CD3+ T lymphocyte ratio showed significant increase in male piglets(P0.05).The proportions of CD8+ T lymphocytes in peripheral blood mononuclear cells at the four time points were(23.80±6.67)%, (17.14±10.64)%, (25.46±12.32)%, (33.33±8.93)% in male piglets and (28.55±6.75)%, (23.62±11.51)%, (37.37±13.18)%, (38.31±15.66)% in female piglets.With the increase of age, the CD8+ T lymphocytes showed a trend of firstly decrease and then increase.The distribution of lymphocyte subsets in weaned piglets showed different changes with the increase of weaning time, and was influenced by gender.The combination of two different cell delineation methods could reflect more accurate changes of the number and proportion of lymphocyte subsets in piglets.%为建立健康仔猪外周血淋巴细胞亚群的临床参考范围,试验分析不同生长阶段仔猪的免疫系统功能水平,分别在健康仔猪断奶后第1、7、21和35天采取外周血,分离外周血单个核细胞,采用三色流式细胞术的方法检测各类主要T淋巴细胞亚群所占外周血单个核细胞的比例,及其占CD3+ T淋巴细胞亚群中的比例,计算CD4+ /CD8+ 的比值.结果表明,随着日龄的增加公母仔猪CD3+ T淋巴细胞亚群占外周血单个核细胞的比例逐渐升高,分别从(38.00±10.94)% 和(38.17±15.39)%上升至(55.43±12.30)%和(51.57±10.60)%.在断奶后1 d和7 d公母仔猪CD3+ T淋巴细胞比例无显著性差异(P>0.05),至断奶后21 d母仔猪CD3+ T淋巴细胞比例显著增加(P0.05).公母仔猪CD8+ T淋巴细胞占外周血单个核细胞的比例在各时间点分别为(23.80±6.67)%、(17.14±10.64)%、(25.46±12.32)%、(33.33±8.93)%和(28.55±6.75)%、(23.62±11.51)%、(37.37±13.18)%、(38.31±15.66)%.随着日龄的增加公母断奶仔猪CD8+ T淋巴细胞呈现先下降后升高的趋势.断奶仔猪各淋巴细胞亚群分布会随着断奶时间的增加而呈现不同的变化,并且存在着性别上的差异.结合两种不同的细胞圈定方法,能够更准确地反映出仔猪体内各淋巴细胞亚群数量和比例的变化.