大鼠感染泡球蚴后外周血及脾脏中滤泡辅助T淋巴细胞的变化

张耀刚; 李建华; 侯静; 孙莉; 田美媛; 江源; 黄登亮; 张涛; 马艳艳; 樊海宁

doi:10.3969/j.issn.1001-5256.2021.07.035

大鼠感染泡球蚴后外周血及脾脏中滤泡辅助T淋巴细胞的变化

DOI: 10.3969/j.issn.1001-5256.2021.07.035

张耀刚^{1, 2},
李建华¹,
侯静¹,
孙莉¹,
田美媛¹,
江源¹,
黄登亮¹,
张涛^{1, 3},
马艳艳^{1, 2, ,},
樊海宁^{1, 2, ,}

1.
青海大学附属医院中心实验室，西宁 810000
2.
青海省包虫病研究重点实验室，西宁 810000
3.
青海大学临床医学院，西宁 810000

基金项目:

青海大学附属医院中青年项目 (ASRF-2018-YB-01);

青海省科技厅重大科技专项 (2016-SF-A5);

国家自然科学基金 (81960129)

利益冲突声明：本研究不存在研究者、伦理委员会成员、受试者监护人以及与公开研究成果有关的利益冲突。

作者贡献声明：马艳艳、樊海宁负责课题设计，拟定写作思路；张耀刚、李建华负责资料分析，撰写论文；侯静、孙莉、田美媛、江源、张涛、黄登亮参与收集数据，修改论文；马艳艳、樊海宁指导撰写文章并最后定稿。

详细信息

通信作者:
马艳艳，mayanyan_research@qhu.edu.cn

樊海宁，fanhaining@medmail.com.cn

中图分类号: R532.32
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- 被引次数: 0
出版历程
- 收稿日期: 2020-12-10
- 录用日期: 2021-01-12
- 出版日期: 2021-07-20

Change in follicular helper T cells in the peripheral blood and spleen of rats after Echinococcus multilocularis infection

Yaogang ZHANG^{1, 2},
Jianhua LI¹,
Jing HOU¹,
Li SUN¹,
Meiyuan TIAN¹,
Yuan JIANG¹,
Dengliang HUANG¹,
Tao ZHANG^{1, 3},
Yanyan MA^{1, 2
, ,},
Haining FAN^{1, 2
, ,}

1.
Central Laboratory of Qinghai University Affiliated Hospital, Xining 810000, China
2.
Qinghai Province Key Laboratory of Hydatid Disease Research, Xining 810000, China
3.
School of Clinical Medicine, Qinghai University, Xining 810000, China

Research funding:

Qinghai University Affiliated Hospital for Young and Middle-aged Project (ASRF-2018-YB-01);

Qinghai Provincial Department of Science and Technology Major Science and Technology Project (2016-SF-A5);

National Natural Science Foundation of China (81960129)

摘要

摘要: 目的分析泡球蚴感染宿主后外周血和脾脏中滤泡辅助T淋巴细胞(Tfh)水平与包虫病进展的关系。方法 20只SD大鼠随机分为正常对照组和模型组，每组10只，模型组开腹直视下在右肝接种约2000个原头节，对照组不做任何处理，3个月后麻醉处死SD大鼠，取外周血和脾脏细胞，以CD4⁺CXCR5⁺PD1⁺为Tfh细胞的标记，使用流式细胞术检测外周血和脾脏中Tfh细胞的水平。使用t检验比较两组之间Tfh细胞的差异。结果大鼠感染泡球蚴3个月后，肝脏可见明显病灶，HE染色显示病灶中可见原头节。泡球蚴感染模型组外周血中CD4⁺CXCR5⁺PD1⁺Tfh细胞在CD4⁺细胞中的平均占比为25.63%±3.47%，正常对照组为11.12%±2.94%，2组比较差异有统计学意义(t=10.230，P＜0.001)，模型组CD4⁺CXCR5⁺PD1⁺Tfh细胞在所有细胞中所占的比例较正常对照组下降显著(0.08%±0.02% vs 0.18%±0.05%, t=5.520，P＜0.001)；在所有细胞中，模型组脾脏中CD4⁺CXCR5⁺PD1⁺Tfh细胞所占的比例(3.00%±0.42%)显著低于正常对照组(5.30%±1.40%)(t=4.769，P＜0.001)。结论外周血中Tfh细胞与包虫病进展密切相关，有望成为反应泡球蚴感染的指标。
- 棘球蚴病, 肝 /
- T淋巴细胞 /
- 大鼠, Sprague-Dawley
Abstract: Objective To investigate the level of follicular helper T (Tfh) cells in the peripheral blood and spleen of the host after Echinococcus multilocularis infection and its association with the progression of echinococcosis. Methods A total of 20 Sprague-Dawley rats were randomly divided into normal control group and model group, with 10 rats in each group. The rats in the model group were inoculated with about 2000 protoscoleces in the right liver under direct-view laparotomy, and those in the control group were not given any treatment. The rats were anesthetized and sacrificed after 3 months to collect peripheral blood and spleen cells, and with CD4⁺CXCR5⁺PD1⁺ as the marker of Tfh cells, flow cytometry was used to measure the level of Tfh cells in peripheral blood and spleen. The t-test was used for comparison of Tfh cells between the two groups. Results After 3 months of Echinococcus multilocularis infection, marked lesions were observed in the liver, and HE staining showed the presence of protoscoleces in the lesions. The proportion of CD4⁺CXCR5⁺PD1⁺Tfh cells in CD4⁺ cells in peripheral blood was 25.63%±3.47% in the model group and 11.12%±2.94% in the normal control group (t=10.230, P < 0.001), a nd the model group had a significantly lower proportion of CD4⁺CXCR5⁺PD1⁺Tfh cells in all cells than the normal control group (0.08%±0.02% vs 0.18%±0.05%, t=5.520, P < 0.001). For the model group, the proportion of CD4⁺CXCR5⁺PD1⁺Tfh cells in all cells in the spleen decreased to 3.00%±0.42%, which was significantly lower than the proportion of 5.30%±1.40% in the normal control group (t=4.769, P < 0.001). Conclusion Tfh cells in peripheral blood are closely associated with the progression of echinococcosis and are expected to become an indicator of Echinococcus multilocularis infection.
- Echinococcosis, Hepatic /
- T-Lymphocytes /
- Rats, Sprague-Dawley

HTML全文

图 1 模型建造情况

注：a, 感染多房棘球蚴3个月后，SD大鼠肝脏病灶情况；b, 病灶HE染色结果(×100)。

下载: 全尺寸图片幻灯片

图 2 AE模型鼠外周血中Tfh细胞的流式细胞检测结果

注：空白组，未标记荧光抗体用于流式细胞术圈门设定界限的组；a、d、g, 外周血中淋巴细胞；b、e、h, CD4⁺细胞；c、f、i，CD4⁺中CXCR5和PD1阳性细胞，其中Q2门表示CD4⁺CXCR5⁺PD1⁺Tfh细胞。

下载: 全尺寸图片幻灯片

图 3 AE模型鼠脾脏中Tfh细胞的流式细胞检测结果

注：a、d、g, 目的细胞群；b、e、h, CD4⁺细胞；c、f、i, CD4⁺中CXCR5和PD1阳性细胞, 其中Q2门表示CD4⁺CXCR5⁺PD1⁺Tfh细胞。

下载: 全尺寸图片幻灯片

表 1 AE模型鼠外周血中Tfh细胞表达情况

组别	动物数(只)	Tfh细胞在CD4⁺细胞中百分比(%)	Tfh细胞在所有细胞中百分比(%)
正常对照组	10	11.12±2.94	0.18±0.05
模型组	10	25.63±3.47	0.08±0.02
t值		10.230	5.520
P值		＜0.001	＜0.001

下载: 导出CSV

表 2 AE模型鼠脾脏中Tfh细胞表达情况

组别	动物数(只)	Tfh细胞在CD4⁺细胞中百分比(%)	Tfh细胞在所有细胞中百分比(%)
正常对照组	10	35.82±4.13	5.30±1.40
模型组	10	33.50±6.40	3.00±0.42
t值		0.974	4.769
P值		0.343	＜0.001

下载: 导出CSV

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