PD-1-mediated CD4<sup>+</sup> T cell exhaustion exacerbates gut microbiota translocation in mouse model of sepsis

QING Bin; KONG Xinxin; YE Dongfan; WANG Chuangye; ZHANG Jian; WANG Bin; HUANG Xiaoou; WANG Nanbo; QIAN Hang; XU Zhi

doi:10.16016/j.2097-0927.202507039

您当前的位置：

首页 >

文章列表页 >

PD-1-mediated CD4⁺ T cell exhaustion exacerbates gut microbiota translocation in mouse model of sepsis

Military Medicine | 更新时间：2025-10-13

- PD-1-mediated CD4⁺ T cell exhaustion exacerbates gut microbiota translocation in mouse model of sepsis
- Journal of Army Medical University Vol. 47, Issue 19, Pages: 2302-2314(2025)
- 作者机构：
  
  1.陆军军医大学（第三军医大学）第二附属医院呼吸与危重症医学科，重大呼吸疾病精准诊疗与防控重庆市重点实验室，呼吸疾病研究重庆市高校重点实验室，重庆
  2.渝粤病理科学研究中心，重庆
- 作者简介：
- 基金信息：
  
  the Fund for Priority Support Targets of Army Medical University(2019R025);the Basic Research Project of Army Medical University(2020D048);the Joint Innovation and Development Fund of Natural Science Foundation of Chongqing(CSTB 2023NSCQ-LZX0165)
- DOI：10.16016/j.2097-0927.202507039
  CLC： R378.2;R392.32;R631.2
- Received：12 July 2025，
  
  Revised：2025-09-12，
  
  Accepted：12 September 2025，
  
  Published：15 October 2025
- 稿件说明：
移动端阅览
卿斌，孔鑫鑫，叶东樊，等.PD-1介导的CD4⁺T细胞耗竭加剧脓毒症小鼠的肠道菌群异位［J］.陆军军医大学学报，2025，47（19）：2302-2314.DOI：10.16016/j.2097-0927.202507039.

QING B，KONG X X，YE D F，et al.PD-1-mediated CD4⁺ T cell exhaustion exacerbates gut microbiota translocation in mouse model of sepsis［J］.J Army Med Univ，2025，47（19）：2302-2314.DOI：10.16016/j.2097-0927.202507039.
卿斌，孔鑫鑫，叶东樊，等.PD-1介导的CD4⁺T细胞耗竭加剧脓毒症小鼠的肠道菌群异位［J］.陆军军医大学学报，2025，47（19）：2302-2314.DOI：10.16016/j.2097-0927.202507039. DOI：

QING B，KONG X X，YE D F，et al.PD-1-mediated CD4⁺ T cell exhaustion exacerbates gut microbiota translocation in mouse model of sepsis［J］.J Army Med Univ，2025，47（19）：2302-2314.DOI：10.16016/j.2097-0927.202507039. DOI：

摘要

目的

探究脓毒症中免疫耗竭的特征并分析其与肠道菌群异位的相关性。

方法

采用随机数字表法将130只小鼠分为盲肠穿刺结扎（cecal ligation and puncture，CLP）组（

=100）和假手术（sham）组（

=30），通过CLP建立小鼠脓毒症模型；流式细胞术检测分析小鼠外周血CD4

T、CD8

T和PD-1

T细胞亚群比例变化；心、肝、肾等脏器匀浆涂板检测细菌定植量变化；HE染色分析免疫器官的病理改变；免疫组化染色检测免疫器官内CD4、CD8和PD-1阳性细胞表达及定位，使用Image J软件对阳性细胞数变化进行定量。

结果

HE染色提示各级免疫器官随病程进展出现不同程度病理损伤；与Sham组小鼠相比，CLP组小鼠各脏器及外周血细菌定植量均显著增高（

0.05），以肝脏感染最为严重；CLP组小鼠在术后第1、3、5天时外周血CD4

T淋巴细胞比例较Sham组分别下降56%、70.57%和87.42%（

0.001），CD8

T淋巴细胞比例只在第5 天时下降48.33%（

0.001），而表达PD-1的CD4

T细胞亚群比例分别升高为Sham组的673.08倍、423.08倍和600倍（

0.001）；免疫组化结果显示，CLP术后胸腺、脾脏和肠系膜淋巴结中CD4阳性细胞比例在早期最高时分别上升为Sham组的7.65倍、2.66倍和3.7倍（

0.001），后期最低时分别较早期下降82.8%（

0.001）、41.9%（

0.01）和60.15%（

0.001）；CD8阳性细胞比例在早期升高后下降不显著；PD-1阳性细胞占比持续升高，最高时分别上升为Sham组的6.24倍、13.9倍和20.96倍（

0.001），且其表达区域与CD4阳性细胞大致相同。

结论

脓毒症时炎症反应可引起免疫器官严重损伤，CD4

T淋巴细胞持续耗竭，机体预防感染的能力下降，可能是造成肠

道菌群异位加重，导致全身感染的重要原因。

Abstract

Objective

To investigate the characteristics of immune exhaustion in sepsis and analyze its association with gut microbiota translocation.

Methods

A total of 130 mice were randomly divided into a cecal ligation and puncture (CLP) group (

=100) and a Sham group (

=30) Mouse model of sepsis was established with CLP procedure. Flow cytometry was used to analyze the proportions of peripheral blood CD4

T and CD8

T cells and programmed cell death protein 1 (PD-1) positive T cell subsets in mice. Bacterial colonization in organs such as the heart

liver and kidneys was quantified by plating homogenates of the organs. Pathological changes in immune organs were observed with HE staining. The expression and localization of CD4⁺

CD8⁺

and PD-1⁺cells in immune organs were detected with immunohistochemical staining

and Image J software was employed for subsequent quantification of the number of the positive cells.

Results

HE staining demonstrated that immune organs exhibited varying degrees of pathological damages with disease progression. Compared with the Sham mice

the CLP mice exhibited significantly increased bacterial colonization in parenchymal organs and peripheral blood (

0.05)

notably in the liver

which showed the most severe infection. In the CLP group

the proportion of CD4

T lymphocytes in peripheral blood at days 1

and 5 postoperatively was decreased by 56%

70.57%

and 87.42%

respectively

when compared with the Sham group (

0.001). The proportion of CD8

T lymphocytes was decreased by 48.33% relative to the Sham group only at day 5 (

0.001). In contrast

the proportion of CD4

T cell subsets expressing PD-1 was increased to 673.08

423.08

and 600 times that of the Sham g

roup

respectively

at the same postoperative time points (

0.001). Immunohistochemical results showed that

in the CLP group

the proportion of CD4

T cells in the thymus

spleen

and mesenteric lymph nodes was increased to 7.65

2.66

and 3.7 times that of the Sham group

respectively

at the early-stage peak (

0.001)

and then these proportions were decreased by 82.8% (

0.001)

41.9% (

0.01)

and 60.15% (

0.001)

respectively

at the late-stage trough when compared with the early-stage peak in the corresponding organs. The proportion of CD8

positive cells was increased in the early stage and then decreased insignificantly

while the proportion of PD-1

positive cells was increased continuously

and reached 6.24

13.9

and 20.96 times that of the Sham group at the peak in the thymus

spleen

and mesenteric lymph nodes respectively (

0.001)

with their expression regions showing a rough overlap with those of CD4

cells.

Conclusion

During sepsis

the inflammatory response can cause severe damage to immune organs and persistent exhaustion of CD4⁺T lymphocytes

leading to declined defenses against infection

which may be the main causes for exacerbated gut microbiota translocation and then systemic infection.

关键词

Keywords

references

YANG S ， GUO J ， KONG Z ， et al . Causal effects of gut microbiota on sepsis and sepsis-related death： insights from genome-wide Mendelian randomization， single-cell RNA， bulk RNA sequencing， and network pharmacology ［J］. J Transl Med ， 2024 ， 22 （ 1 ）： 10 . DOI： 10.1186/s12967-023-04835-8 http://dx.doi.org/10.1186/s12967-023-04835-8 .

ROSERO O ， KOVÁCS T ， ONODY P ， et al . Bacterial translocation： gap in the shield ［J］. Orv Hetil ， 2014 ， 155 （ 8 ）： 304 - 312 . DOI： 10.1556/OH.2014.29836 http://dx.doi.org/10.1556/OH.2014.29836 .

WOZNIAK H ， BECKMANN T S ， FRÖHLICH L ， et al . The central and biodynamic role of gut microbiota in critically ill patients ［J］. Crit Care ， 2022 ， 26 （ 1 ）： 250 . DOI： 10.1186/s13054-022-04127-5 http://dx.doi.org/10.1186/s13054-022-04127-5 .

CHEN Y F ， SUN K Y ， QI Y ， et al . L-valine derived from the gut microbiota protects sepsis-induced intestinal injury and negatively correlates with the severity of sepsis ［J］. Front Immunol ， 2024 ， 15 ： 1424332 . DOI： 10.3389/fimmu.2024. 1424332 http://dx.doi.org/10.3389/fimmu.2024.1424332 .

VAISHNAVI C . Translocation of gut flora and its role in sepsis ［J］. Indian J Med Microbiol ， 2013 ， 31 （ 4 ）： 334 - 342 . DOI： 10.4103/0255-0857.118870 http://dx.doi.org/10.4103/0255-0857.118870 .

LI C X ， LIU Y ， ZHANG Y Z ， et al . Astragalus polysaccharide： a review of its immunomodulatory effect ［J］. Arch Pharm Res ， 2022 ， 45 （ 6 ）： 367 - 389 . DOI： 10.1007/s12272-022-01393-3 http://dx.doi.org/10.1007/s12272-022-01393-3 .

PFORTMUELLER C A ， MEISEL C ， FUX M ， et al . Assessment of immune organ dysfunction in critical illness： utility of innate immune response markers ［J］. Intensive Care Med Exp ， 2017 ， 5 （ 1 ）： 49 . DOI： 10.1186/s40635-017-0163-0 http://dx.doi.org/10.1186/s40635-017-0163-0 .

NIU M W ， CHEN P . Crosstalk between gut microbiota and sepsis ［J］. Burns Trauma ， 2021 ， 9 ： tkab036 . DOI： 10.1093/burnst/tkab036 http://dx.doi.org/10.1093/burnst/tkab036 .

ZHANG X Y ， ZHANG Y J ， YUAN S Y ， et al . The potential immunological mechanisms of sepsis ［J］. Front Immunol ， 2024 ， 15 ： 1434688 . DOI： 10.3389/fimmu.2024.1434688 http://dx.doi.org/10.3389/fimmu.2024.1434688 .

PEPPER M ， JENKINS M K . Origins of CD4 + effector and central memory T cells ［J］. Nat Immunol ， 2011 ， 12 （ 6 ）： 467 - 471 . DOI： 10.1038/ni.2038 http://dx.doi.org/10.1038/ni.2038 .

FIFE B T ， BLUESTONE J A . Control of peripheral T-cell tolerance and autoimmunity via the CTLA-4 and PD-1 pathways ［J］. Immunol Rev ， 2008 ， 224 ： 166 - 182 . DOI： 10.1111/j.1600-065X.2008.00662.x http://dx.doi.org/10.1111/j.1600-065X.2008.00662.x .

HOTCHKISS R S ， MONNERET G ， PAYEN D . Immunosuppression in sepsis： a novel understanding of the disorder and a new therapeutic approach ［J］. Lancet Infect Dis ， 2013 ， 13 （ 3 ）： 260 - 268 . DOI： 10.1016/S1473-3099（13）70001-X http://dx.doi.org/10.1016/S1473-3099（13）70001-X .

RITTIRSCH D ， HUBER-LANG M S ， FLIERL M A ， et al . Immunodesign of experimental sepsis by cecal ligation and puncture ［J］. Nat Protoc ， 2008 ， 4 （ 1 ）： 31 - 36 . DOI： 10.1038/nprot.2008.214 http://dx.doi.org/10.1038/nprot.2008.214 .

BOOMER J S ， TO K ， CHANG K C ， et al . Immunosuppression in patients who die of sepsis and multiple organ failure ［J］. JAMA ， 2011 ， 306 （ 23 ）： 2594 - 2605 . DOI： 10.1001/jama.2011.1829 http://dx.doi.org/10.1001/jama.2011.1829 .

SINGER M ， DEUTSCHMAN C S ， SEYMOUR C W ， et al . The third international consensus definitions for sepsis and septic shock （sepsis-3）［J］. JAMA ， 2016 ， 315 （ 8 ）： 801 - 810 . DOI： 10.1001/jama.2016.0287 http://dx.doi.org/10.1001/jama.2016.0287 .

XIE J F ， WANG H L ， KANG Y ， et al . The epidemiology of sepsis in Chinese ICUs： a national cross-sectional survey ［J］. Crit Care Med ， 2020 ， 48 （ 3 ）： e209 - e218 . DOI： 10.1097/CCM.0000000000004155 http://dx.doi.org/10.1097/CCM.0000000000004155 .

KAMADA N ， SEO S U ， CHEN G Y ， et al . Role of the gut microbiota in immunity and inflammatory disease ［J］. Nat Rev Immunol ， 2013 ， 13 （ 5 ）： 321 - 335 . DOI： 10.1038/nri3430 http://dx.doi.org/10.1038/nri3430 .

BASU S . Neonatal sepsis： the gut connection ［J］. Eur J Clin Microbiol Infect Dis ， 2015 ， 34 （ 2 ）： 215 - 222 . DOI： 10.1007/s10096-014-2232-6 http://dx.doi.org/10.1007/s10096-014-2232-6 .

PROTZER U ， MAINI M K ， KNOLLE P A . Living in the liver： hepatic infections ［J］. Nat Rev Immunol ， 2012 ， 12 （ 3 ）： 201 - 213 . DOI： 10.1038/nri3169 http://dx.doi.org/10.1038/nri3169 .

SUN J ， ZHANG J X ， WANG X F ， et al . Gut-liver crosstalk in sepsis-induced liver injury ［J］. Crit Care ， 2020 ， 24 （ 1 ）： 614 . DOI： 10.1186/s13054-020-03327-1 http://dx.doi.org/10.1186/s13054-020-03327-1 .

VINCENT J L ， ANGUS D C ， ARTIGAS A ， et al . Effects of drotrecogin Alfa （activated） on organ dysfunction in the PROWESS trial ［J］. Crit Care Med ， 2003 ， 31 （ 3 ）： 834 - 840 . DOI： 10.1097/01.CCM.0000051515.56179.E1 http://dx.doi.org/10.1097/01.CCM.0000051515.56179.E1 .

ASSIMAKOPOULOS S F ， TRIANTOS C ， THOMOPOULOS K ， et al . Gut-origin sepsis in the critically ill patient： pathophysiology and treatment ［J］. Infection ， 2018 ， 46 （ 6 ）： 751 - 760 . DOI： 10.1007/s15010-018-1178-5 http://dx.doi.org/10.1007/s15010-018-1178-5 .

SHIVE C L ， KOWAL C M ， DESOTELLE A F ， et al . Endotoxemia associated with liver disease correlates with systemic inflammation and T cell exhaustion in hepatitis C virus infection ［J］. Cells ， 2023 ， 12 （ 16 ）： 2034 . DOI： 10.3390/cells12162034 http://dx.doi.org/10.3390/cells12162034 .

CHEN J X ， CHEN R Y ， HUANG S X ， et al . Atezolizumab alleviates the immunosuppression induced by PD-L1-positive neutrophils and improves the survival of mice during sepsis ［J］. Mol Med Rep ， 2021 ， 23 （ 2 ）： 144 . DOI： 10.3892/mmr.2020.11783 http://dx.doi.org/10.3892/mmr.2020.11783 .

THAPA P ， FARBER D L . The role of the Thymus in the immune response ［J］. Thorac Surg Clin ， 2019 ， 29 （ 2 ）： 123 - 131 . DOI： 10.1016/j.thorsurg.2018.12.001 http://dx.doi.org/10.1016/j.thorsurg.2018.12.001 .

KUCHLER L ， SHA L K ， GIEGERICH A K ， et al . Elevated intrathymic sphingosine-1-phosphate promotes Thymus involution during sepsis ［J］. Mol Immunol ， 2017 ， 90 ： 255 - 263 . DOI： 10.1016/j.molimm.2017.08.011 http://dx.doi.org/10.1016/j.molimm.2017.08.011 .

CESTA M F . Normal structure， function， and histology of the spleen ［J］. Toxicol Pathol ， 2006 ， 34 （ 5 ）： 455 - 465 . DOI： 10.1080/01926230600867743 http://dx.doi.org/10.1080/01926230600867743 .

CRANE G M ， LIU Y C ， CHADBURN A . Spleen： Development， anatomy and reactive lymphoid proliferations ［J］. Semin Diagn Pathol ， 2021 ， 38 （ 2 ）： 112 - 124 . DOI： 10.1053/j.semdp.2020.06.003 http://dx.doi.org/10.1053/j.semdp.2020.06.003 .

SENTHIL M ， WATKINS A ， BARLOS D ， et al . Intravenous injection of trauma-hemorrhagic shock mesenteric lymph causes lung injury that is dependent upon activation of the inducible nitric oxide synthase pathway ［J］. Ann Surg ， 2007 ， 246 （ 5 ）： 822 - 830 . DOI： 10.1097/SLA.0b013e3180caa3af http://dx.doi.org/10.1097/SLA.0b013e3180caa3af .

RUIZ S ， VARDON-BOUNES F ， MERLET-DUPUY V ， et al . Sepsis modeling in mice： ligation length is a major severity factor in cecal ligation and puncture ［J］. Intensive Care Med Exp ， 2016 ， 4 （ 1 ）： 22 . DOI： 10.1186/s40635-016-0096-z http://dx.doi.org/10.1186/s40635-016-0096-z .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

ATF3 regulates macrophage autophagy and inflammatory responses by suppressing ATG5 and ATG16L1 expression

Preparation of Trop2-targeted chimeric antigen receptor T cells and study of their exhaustion status in vitro

Protective effect of remimazolam on intestinal barrier function in septic mice

Dexmedetomidine pretreatment enhances protective effect efficacy of pericytes agaist acute lung injury in septic mice

Related Author

WANG Yujie

QIU Hongmei

YANG Ting

SHI Xinhui

YANG Xiong

JIANG Qingsong

LIU Xin

LI Xiaoli

Related Institution

Clinical Research Center， First Affiliated Hospital， Army Medical University （Third Military Medical University）

Faculty of Pharmacology， College of Pharmacy， Chongqing Medical University

Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology

Dezhou Health Commission

College of Life Sciences， Beijing University of Chinese Medicine

⁰

PD-1-mediated CD4+ T cell exhaustion exacerbates gut microbiota translocation in mouse model of sepsis

DOI：10.16016/j.2097-0927.202507039

摘要

Abstract

关键词

Keywords

references

PD-1-mediated CD4⁺ T cell exhaustion exacerbates gut microbiota translocation in mouse model of sepsis