不同日龄仔猪外周血T淋巴细胞的检测与分析

doi:10.3969/j.issn.1004-1524.2017.08.05

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (8): 1253-1260.DOI: 10.3969/j.issn.1004-1524.2017.08.05

不同日龄仔猪外周血T淋巴细胞的检测与分析

邹璐^{1, 2}, 张景艳², 张凯², 王磊², 张康², 王旭荣², 王学智², 余四九¹, 杨志强^{2, *}, 李建喜^{2, *}

1.甘肃农业大学动物医学院, 甘肃兰州 730050；
2.中国农业科学院兰州畜牧与兽药研究所,甘肃省中兽药工程技术研究中心,甘肃兰州 730050

收稿日期:2016-12-30 出版日期:2017-08-20 发布日期:2017-09-06
通讯作者: 杨志强, E-mail: zqycaas@126.com;李建喜,E-mail: lijianxigs@126.com
作者简介:邹璐(1990-),男,河南潢川人,硕士研究生,主要从事中兽药的研究。E-mail: 1406832500@qq.com
基金资助:
公益性行业专项(201303040-15); 国家自然科学基金面上项目(31472233); 甘肃省科技支撑计划(1604NKCA069-03)

Analysis of T lymphocytes in peripheral blood of piglets in different growth stages

ZOU Lu^{1, 2}, ZHANG Jingyan², ZHANG Kai², WANG Lei², ZHANG Kang², WANG Xurong², WANG Xuezhi², YU Sijiu¹, YANG Zhiqiang^{2, *}, LI Jianxi^{2, *}

1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730050, China;
2. Engineering & Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Science of CAAS , Lanzhou 730050, China;

Received:2016-12-30 Online:2017-08-20 Published:2017-09-06

摘要/Abstract

摘要： 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(P<0.05),公仔猪至35 d时CD3⁺ T淋巴细胞比例显著增加(P<0.05)。随着日龄的增长,公母仔猪CD4⁺ T淋巴细胞亚群占外周血单个核细胞的比例分别由(10.29±2.71)% 和(10.3±2.17)%上升至(22.2±5.87)%和(23.4±6.84)%。公母仔猪CD4⁺ T淋巴细胞的比例逐渐升高,且各个时间点公母仔猪结果无明显差异(P>0.05)。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。

关键词: 仔猪, T淋巴细胞亚群, 流式细胞术, 细胞圈定

Abstract: 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(P<0.05). On the 35th day, the percentage of CD3⁺ T cells showed a significant increase (P<0.05). With the increase of age, the proportion of CD4⁺ T lymphocyte subsets in peripheral blood mononuclear cells of male and female piglets increased from (10.29±2.71)% to( 22.2±5.87)% and from (10.3±2.17)% to (23.4±6.84)%, respectively. The proportions of CD4⁺ T lymphocytes in male and female piglets were gradually increasing, and there was no significant difference between male and female piglets at different time points(P>0.05). 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.

Key words: piglets, T lymphocyte subsets, flow cytometry, cell delineation

中图分类号:

S811.2

邹璐, 张景艳, 张凯, 王磊, 张康, 王旭荣, 王学智, 余四九, 杨志强, 李建喜. 不同日龄仔猪外周血T淋巴细胞的检测与分析[J]. 浙江农业学报, 2017, 29(8): 1253-1260.

ZOU Lu, ZHANG Jingyan, ZHANG Kai, WANG Lei, ZHANG Kang, WANG Xurong, WANG Xuezhi, YU Sijiu, YANG Zhiqiang, LI Jianxi. Analysis of T lymphocytes in peripheral blood of piglets in different growth stages[J]. , 2017, 29(8): 1253-1260.

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不同日龄仔猪外周血T淋巴细胞的检测与分析

Analysis of T lymphocytes in peripheral blood of piglets in different growth stages

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