Distortion of memory Vδ2 γδ T cells contributes to immune dysfunction in chronic HIV infection

γδT cells play important roles in innate immunity as the first-line of defense against infectious diseases. Human immunodeficiency virus （HIV） infection disrupts the balance between Vδ1 T cells and Vδ2 T cells and causes dysfunction among γδ T cells. However, the biological mechanisms and clinical consequences of this disruption require further investigation. In this study, we performed a comprehensive analysis of phenotype and function of memory γδ T cells in cohorts of Chinese individuals with HIV infection. We found a dynamic change in memory Vδ2 γδ T cells, skewed toward an activated and terminally differentiated effector memory phenotype TEMRA Vδ2 γδT cell, which may account for the dysfunction of Vδ2 γδT cells in HIV disease. In addition, we found that IL-17-producing γδ T cells were significantly increased in HIV-infected patients with fast disease progression and positively correlated with HLA-DR＋ γδ T cells and CD38＋HLA-DR＋ γδ T cells. This suggests the IL-17 signaling pathway is involved in γδ T-cell activation and HIV pathogenesis. Our findings provide novel insights into the role of Vδ2 T cells during HIV pathogenesis and represent a sound basis on which to consider immune therapies with these cells.

肿瘤相关性贫血与SARS-CoV-2特异性记忆CD8^(+)T细胞丰度的相关性分析

目的 探讨肿瘤相关性贫血对外周血中新型冠状病毒特异性记忆CD8^(+)T细胞丰度的影响。方法 纳入2023年6-7月在火箭军特色医学中心住院治疗的HLA-A*11∶01阳性中晚期实体瘤患者34例。根据血红蛋白浓度将患者分为无贫血组(HGB≥120 g/L,n=14)、轻度贫血组(120 g/L> HGB≥90 g/L,n=14)与中重度贫血组(HGB<90 g/L,n=6)。分离外周血单个核细胞,利用新冠病毒抗原表位刺激并扩增记忆CD8^(+)T细胞,使用ELISPOT筛选阳性表位,构建Tetramer。利用扩增后细胞检测Tetramer+细胞比例、数量及ELISPOT斑点数,用以反映新冠病毒特异性记忆CD8^(+)T细胞丰度。分析患者血红蛋白浓度与Tetramer+细胞比例、数量及ELISPOT斑点数的相关性。分析不同贫血组Tetramer+细胞比例、数量及ELISPOT斑点数的差异。结果 34例患者中男性25例,女性9例;年龄36~78岁,中位年龄57岁。患者扩增后Tetramer+细胞比例、数量及ELISPOT斑点数与血红蛋白浓度均呈正相关关系(P<0.05);与无贫血组相比,中重度贫血组患者Tetramer+细胞比例、数量及ELISPOT斑点数均明显降低(P<0.05)。结论 肿瘤患者外周血新冠病毒特异性记忆CD8^(+)T细胞的丰度与血红蛋白浓度正相关,提示肿瘤相关性贫血可能通过降低外周血中记忆CD8^(+)T细胞的丰度导致患者病毒易感性增高。

免疫检查点T细胞激活抑制物免疫球蛋白可变区结构域、人内源性逆转录病毒-H长端重复相关蛋白2在子宫内膜癌中的表达及临床意义

目的分析免疫检查点T细胞激活抑制物免疫球蛋白可变区结构域(VISTA)、人内源性逆转录病毒-H长端重复相关蛋白2(HHLA2)在子宫内膜癌(EC)中的表达及临床意义。方法选取2017年9月至2019年10月唐山市妇幼保健院收治的120例的EC患者作为研究对象,收集其的EC组织和癌旁正常组织。检测EC组织和癌旁正常组织VISTA、HHLA2的表达;采用Spearman分析免疫检查点VISTA、HHLA2的相关性;Kaplan-Meier分析VISTA、HHLA2与预后的关系;Cox回归分析影响EC患者预后的危险因素。结果EC组织中VISTA、HHLA2阳性表达率显著高于正常癌旁组织(P<0.05)。Spearman相关性结果显示EC组织中VISTA与HHLA2表达呈正相关性(r=0.587,P<0.05)。EC组织中VISTA、HHLA2表达与临床病理特征有关(P<0.05)。Kaplan-Meier曲线结果显示VISTA阳性表达患者3年生存率低于阴性表达患者(χ^(2)=11.864,P<0.05),HHLA2阳性表达患者3年生存率低于阴性表达患者(χ^(2)=4.975,P<0.05)。Cox回归分析结果显示VISTA和HHLA2是影响EC患者预后的危险因素(P<0.05)。结论免疫检查点VISTA、HHLA2在EC中高表达,其是影响EC预后的危险因素,二者可能是有价值的预后标志物。

健康个体中SARS-CoV-2反应性记忆T细胞免疫研究

目的 了解健康个体中的SARS-CoV-2反应性CD4+T细胞的记忆表型和功能。方法 使用SARS-CoV-2来源多肽刺激来自参与者的PBMC,通过胞内染色和流式细胞术检测SARS-CoV-2反应性记忆T细胞,并进行记忆表型分析,通过CBA检测SARS-CoV-2来源多肽刺激后的细胞因子分泌水平,初步评估SARS-CoV-2反应性记忆T细胞的功能。结果 40%(6/15)健康个体可检测到SARS-CoV-2反应性CD4+记忆T细胞,记忆表型分析显示其主要由中央记忆型T细胞组成(82.2%),其他记忆细胞占17.8%。与阴性对照相比,SARS-CoV-2来源多肽刺激后,IL-10分泌水平的下降具有统计学意义(P<0.05),IFNγ、TNFα、IL-2和IL-4分泌水平的差异不具有统计学意义。结论 健康个体中存在SARS-CoV-2反应性CD4+记忆T细胞。

Comprehensive analysis of endoplasmic reticulum stress-related mechanisms in type 2 diabetes mellitus

BACKGROUND The endoplasmic reticulum(ER)is closely related to a wide range of cellular functions and is a key component to maintain and restore metabolic health.Type 2 diabetes mellitus(T2DM)is a serious threat to human health,but the ER stress(ERS)-related mechanisms in T2DM have not been fully elucidated.AIM To identify potential ERS-related mechanisms and crucial biomarkers in T2DM.METHODS We conducted gene set enrichment analysis(GSEA)and gene set variation analysis(GSVA)in myoblast and myotube form GSE166502,and obtained the differentially expressed genes(DEGs).After intersecting with ERS-related genes,we obtained ERS-related DEGs.Finally,functional analyses,immune infiltration,and several networks were established.RESULTS Through GSEA and GSVA,we identified several metabolic and immune-related pathways.We obtained 227 ERS-related DEGs and constructed several important networks that help to understand the mechanisms and treatment of T2DM.Finally,memory CD4^(+)T cells accounted for the largest proportion of immune cells.CONCLUSION This study revealed ERS-related mechanisms in T2DM,which might contribute to new ideas and insights into the mechanisms and treatment of T2DM.

COVID-19患者体内SARS-CoV-2特异性的记忆性CD8^(+)T细胞和抗体水平的变化

目的研究SARS-CoV-2病人恢复期的病毒特异性的记忆性T细胞和抗体水平的变化。方法通过SARS-CoV-2特异性抗原肽刺激COVID-19病人来源PBMC,流式细胞术检测不同疾病严重程度COVID-19患者SARS-CoV-2特异性的记忆性CD8^(+)T细胞的数目以及功能,ELISA检测不同疾病严重程度COVID-19患者血清中S1、S2特异性的IgG水平。结果与中、重症COVID-19患者相比,无症状或轻症的COVID-19患者体内拥有数目更多,质量更好的SARS-CoV-2特异性的记忆性CD8^(+)T细胞;中、重症SARS-CoV-2感染患者血清中病毒特异性IgG水平随着时间延长而降低,维持时间较短。结论无症状和轻症COVID-19患者体内拥有质量更好的病毒特异性的记忆性CD8^(+)T细胞。

CD3ζ链引入YRHQ基序可增强靶向HER2的CAR-T细胞的抗肿瘤活性

目的:探讨在靶向HER2的CAR的CD3ζ链胞内区引入YRHQ基序对CAR-T细胞的特异性杀伤活性及免疫记忆形成的影响。方法:通过DNA合成获得包含靶向HER2的编码抗原受体H28ζ或H28ζ(YRHQ)的DNA片段,通过慢病毒载体将不同CAR的DNA片段分别转导健康人外周血T细胞,制备靶向HER2的H28ζ-CAR-T及H28ζ(YRHQ)-CAR-T细胞。扩增过程中对不同CAR-T细胞进行计数,FCM检测CAR的表达率。将CAR-T细胞分别与HER2阳性的SKOV3、MDA-MB-453或HER2阴性的MCF-7细胞共培养,LDH释放法检测其杀伤活性,ELISA法检测IL-2、IFN-γ和颗粒酶B的水平,WB法检测STAT3磷酸化水平及免疫检查点分子TIM-3和PD-1的表达,通过FCM检测CCR7、CD45RO的表达,分析CAR-T细胞的表型。结果:H28ζ-CAR-T和H28ζ(YRHQ)-CAR-T细胞扩增能力较好,体外培养7 d时扩增4~5倍。H28ζ-CAR和H28ζ(YRHQ)-CAR表达率分别为(33.3±2.85)%和(28.30±3.2)%。H28ξ(YRHQ)-CAR-T细胞的杀伤活性较H28ζ-CAR-T细胞更高(P<0.05)。经HER2抗原刺激后,与T细胞或H28z-CAR-T细胞比较,H28ξ(YRHQ)-CAR-T细胞的STAT3磷酸化水平较H28ξ-CAR-T细胞明显升高(P<0.01);而两者间PD-1和TIM-3的表达无明显差异。未经抗原刺激的CAR-T细胞CCR7和CD45RO表达与正常T细胞比较差异无统计学意义(均P>0.05),与SKOV3细胞共培养后,与T细胞或H28z-CAR-T细胞比较,H28ξ(YRHQ)-CAR-T细胞中T_(EM)细胞比例明显增加、T_(CM)细胞比例明显减少(均P<0.05)。结论:在CD3胞内区引入YRHQ基序可在一定程度上提高CAR-T细胞的杀伤潜力。

Antigen-presenting effects of effector memory Vγ9Vδ2 T cells in rheumatoid arthritis

Rheumatoid arthritis is an autoimmune disease that primarily affects the limbs, but the pathogenic mechanism remains unclear. 78 T cells, a T-cell subpopulation, are characterized by multiple biological functions and associated with a variety of diseases. This study investigated the antigen-presenting effects of γδ2 cells and their relationship with rheumatoid arthritis development. We found that Vγ9Vδ2 T cells （the predominant subtype of γδ T cells in peripheral blood） were activated by isopentenyl pyrophosphate to continuously proliferate and differentiate into effector memory cells. The effector memory Vγ9Vδ2 T cells exhibited phenotypic characteristics of specific antigen-presenting cells, including high HLA-DR and CD80/86 expression. These Vγ9Vδ2 T cells could present soluble antigens and synthetic peptides to CD4＋ T cells. Vγ9Vδ2 T cells with different phenotypes showed different cytokine secretion patterns. Effector memoryVγ9Vδ2 T cells simultaneously secreted not only interferon （IFN）-γbut also IL-17. The peripheral blood and joint synovial fluid from RA patients contained numerous heterogeneous γδ T cells that were predominantly effector memory Vγ9Vδ2 T cells with the ability to secrete inflammatory factors. We also found that γδ T cells had a similar antigen-presenting capability to B cells. These results suggest that during the development of rheumatoid arthritis, 78 T cells can aggravate immune dysfunction and produce abnormal immune damage by secreting cytokines and inducing inflammatory cells to participate in synergistic inflammatory responses. Furthermore, γδ T cells can behave similarly to B cells to present viral peptides and autoantigen peptides to CD4＋ T cells, thus sustaining CD4＋ T-cell activation.

TRAF2 regulates T cell immunity by maintaining a Tpl2-ERK survival signaling axis in effector and memory CD8 T cells

Generation and maintenance of antigen-specific effector and memory T cells are central events in immune responses against infections.We show that TNF receptor-associated factor 2(TRAF2)maintains a survival signaling axis in effector and memory CD8 T cells required for immune responses against infections.This signaling axis involves activation of Tpl2 and its downstream kinase ERK by NF-κB-inducing kinase(NIK)and degradation of the proapoptotic factor Bim.NIK mediates Tpl2 activation by stimulating the phosphorylation and degradation of the Tpl2 inhibitor p105.Interestingly,while NIK is required for Tpl2-ERK signaling under normal conditions,uncontrolled NIK activation due to loss of its negative regulator,TRAF2,causes constitutive degradation of p105 and Tpl2,leading to severe defects in ERK activation and effector/memory CD8 T cell survival.Thus,TRAF2 controls a previously unappreciated signaling axis mediating effector/memory CD8 T cell survival and protective immunity.

Mucosal COVID-19 vaccines:Risks,benefits and control of the pandemic

Based on mucosal immunization to promote both mucosal and systemic immune responses,next-generation coronavirus disease 2019(COVID-19)vaccines would be administered intranasally or orally.The goal of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)vaccines is to provide adequate immune protection and avoid severe disease and death.Mucosal vaccine candidates for COVID-19 including vector vaccines,recombinant subunit vaccines and live attenuated vaccines are under development.Furthermore,subunit protein vaccines and virus-vectored vaccines have made substantial progress in preclinical and clinical settings,resulting in SARS-CoV-2 intranasal vaccines based on the previously successfully used nasal vaccines.Additional to their ability to trigger stable,protective immune responses at the sites of pathogenic infection,the development of‘specific’mucosal vaccines targeting coronavirus antigens could be an excellent option for preventing future pandemics.However,their efficacy and safety should be confirmed.

Human Vγ9Vδ2-T cells efficiently kill influenzavirus-infected lung alveolar epithelial cells

γδ-T cells play an indispensable role in host defense against different viruses, including influenza A virus. However, whether these cells have cytotoxic activity against influenza virus-infected lung alveolar epithelial cells and subsequently contribute to virus clearance remains unknown. Using influenza virus-infected A549 cells, human lung alveolar epithelial cells, we investigated the cytotoxic activity of aminobisphosphonate pamidronate （PAM）-expanded human Vγ9Vδ2-T cells and their underlying mechanisms. We found that PAM could selectively activate and expand human Vγ9Vδ2-T cells. PAM-expanded human Vγ9Vδ2-T cells efficiently killed influenza virus-infected lung alveolar epithelial cells and inhibited virus replication. The cytotoxic activity of PAM-expanded Vγ9Vδ2-T cells was dependent on cell-to-cell contact and required NKG2D activation. Perforin-granzyme B, tumor-necrosis factor-related apoptosis-inducing ligand （TRAIL） and Fas-Fas ligand （FasL） pathways were involved in their cytotoxicity. Our study suggests that targeting γδ2-T cells by PAM can potentially offer an alternative option for the treatment of influenza virus.

Contact-dependent delivery of IL-2 by dendritic cells to CD4 T cells in the contraction phase promotes their long-term survival

Common y chain cytokines are important for immune memory formation.Among them,the role of IL-2 remains to be fully explored.It has been suggested that this cytokine is critically needed in the late phase of primary CD4 T cell activation.Lack of IL-2 at this stage sets for a diminished recall response in subsequent challenges.However,as IL-2 peak production is over at this point,the source and the exact mechanism that promotes its production remain elusive.We report here that resting,previously antigen-stimulated CD4 T cells maintain a minimalist response to dendritic cells after their peak activation in vitro.This subtle activation event may be induced by DCs without overt presence of antigen and appears to be stronger if IL-2 comes from the same dendritic cells.This encounter reactivates a miniature IL-2 production and leads a gene expression profile change in these previously activated CD4 T cells.The CD4 T cells so experienced show enhanced reactivation intensity upon secondary challenges later on.Although mostly relying on in vitro evidence,our work may implicate a subtle programing for CD4 T cell survival after primary activation in vivo.

Self-activation of Vγ9Vδ2 T cells by exogenous phosphoantigens involves TCR and butyrophilins

The high cytotoxic activity of Vγ9Vδ2 T lymphocytes against tumor cells makes them useful candidates in anticancer therapies.However,the molecular mechanism of their activation by phosphoantigens(PAgs)is not completely known.Many studies have depicted the mechanism of Vγ9Vδ2 T-cell activation by PAg-sensed accessory cells,such as immune presenting cells or tumor cells.In this study,we demonstrated that pure resting Vγ9Vδ2 T lymphocytes can self-activate through exogenous PAgs,involving their TCR and the butyrophilins BTN3A1 and BTN2A1.This is the first time that these three molecules,concurrently expressed at the plasma membrane of Vγ9Vδ2 T cells,have been shown to be involved together on the same and unique T cell during PAg activation.Moreover,the use of probucol to stimulate the inhibition of this self-activation prompted us to propose that ABCA-1 could be implicated in the transfer of exogenous PAgs inside Vγ9Vδ2 T cells before activating them through membrane clusters formed byγ9TCR,BTN3A1 and BTN2A1.The self-activation of Vγ9Vδ2 T cells,which leads to self-killing,can therefore participate in the failure ofγδT cell-based therapies with exogenous PAgs and should be taken into account.

T1DM患者外周血记忆性T细胞表面PD-1表达水平与其疾病发生发展的初步研究

目的:通过检测1型糖尿病(type 1 diabetes mellitus,T1DM)、2型糖尿病(type 2 diabetes mellitus,T2DM)患者以及健康对照外周血T细胞亚群免疫负调控分子程序性死亡蛋白1(programmed cell death protein 1,PD-1)m RNA相对表达水平和记忆性T细胞(memory T cells,Tm)表面PD-1表达情况,从分子生物学、细胞免疫学等多种角度研究记忆性T细胞PD-1表达异常与以胰岛β细胞破坏为主要机制的T1DM发生发展的相关性。方法:分离T1DM、T2DM患者以及健康对照组外周血单个核细胞(peripheral blood mononuclear cells,PBMCs);从PBMCs中分离出CD4^+T细胞和CD8^+T细胞,使用荧光定量PCR分别检测其细胞内PD-1 m RNA相对表达水平;将PBMCs分别标记荧光抗体CD4-FITC、CD45RO-PE、CCR7-APC、CD8-FITC、PD-1-Per Cp,应用流式细胞术分别检测CD4^+CD45RO^+CCR7^+Tcm细胞、CD4^+CD45RO^+CCR7-Tem细胞、CD8^+CD45RO^+CCR7^+Tcm细胞、CD8^+CD45RO^+CCR7^-Tem细胞表面免疫负调控分子PD-1的表达水平。结果:(1)T1DM组患者外周血中CD4^+T细胞内PD-1 m RNA相对表达水平低于T2DM组和正常对照组,差异有统计学意义;(2)T1DM组患者外周血中CD8^+T细胞内PD-1 m RNA相对表达水平未见明显异常;(3)T1DM组患者外周血中CD4^+CD45RO^+CCR7-Tem细胞亚群与CD4^+CD45RO^+CCR7^+Tcm细胞亚群PD-1表达水平均显著低于正常对照组和T2DM组患者,差异有统计学意义;(4)T1DM组患者外周血中CD8^+CD45RO^+CCR7-Tem细胞亚群与CD8^+CD45RO^+CCR7^+Tcm细胞亚群PD-1表达水平均无明显异常。结论:胰岛β细胞可通过细胞表面PD-L1与CD4^+Tm细胞表面PD-1结合从而负调节后者细胞免疫作用,因此当CD4^+Tm细胞PD-1表达异常而失去负调控作用时,细胞效应则会进一步增强,最终可能通过破坏胰岛β细胞而导致T1DM的发生发展。

Functions of NKG2D in CD8^(+) T cells: an opportunity for immunotherapy

Natural killer group 2 member D(NKG2D)is a type II transmembrane receptor.NKG2D is present on NK cells in both mice and humans,whereas it is constitutively expressed on CD8+T cells in humans but only expressed upon T-cell activation in mice.NKG2D is a promiscuous receptor that recognizes stress-induced surface ligands.In NK cells,NKG2D signaling is sufficient to unleash the killing response;in CD8+T cells,this requires concurrent activation of the T-cell receptor(TCR).In this case,the function of NKG2D is to authenticate the recognition of a stressed target and enhance TCR signaling.CD28 has been established as an archetype provider of costimulation during T-cell priming.It has become apparent,however,that signals from other costimulatory receptors,such as NKG2D,are required for optimal T-cell function outside the priming phase.This review will focus on the similarities and differences between NKG2D and CD28;less well-described characteristics of NKG2D,such as the potential role of NKG2D in CD8+T-cell memory formation,cancer immunity and autoimmunity;and the opportunities for targeting NKG2D in immunotherapy.

γc家族细胞因子对体外培养T细胞表型的影响

目的:探讨γc家族细胞因子对体外培养T细胞表型的影响,为过继免疫治疗体外细胞制备提供实验依据。方法:采用健康人外周静脉血单个核细胞(PBMC),应用尼龙毛柱法、CD3磁珠阳性分选法、CD3磁珠阴性分选法和自然沉降法等4种方法分选T细胞,比较各种方法获取的T细胞纯度、回收率和细胞增殖。用CD3/CD28免疫磁珠激活CD3^+T细胞,γc家族细胞因子分为IL-2组和IL-7、IL-15、IL-21混合组,比较两组T细胞扩增倍数和表型的差异。结果:CD3磁珠阴性分选的T细胞纯度显著高于尼龙毛柱分选、CD3磁珠阳性分选、自然沉降分选[(94.06±1.07)%vs(86.74±1.06)%、(89.61±1.40)%、(88.48±1.86)%,P<0.05],自然沉降分选的T细胞回收率显著高于其余3种分选法[(60.29±1.53)%vs(45.03±2.79)%、(20.15±3.41)%、(42.98±2.82)%,P<0.05),综合比较效果以自然沉降法为最优选择。IL-2组T细胞扩增倍数显著高于混合组[(262.6±143.2)vs(73.0±25.8)倍,P<0.05]。混合组的早期记忆T细胞占总T细胞的比例、T_(scm)和T_(scm-like)、T_(cm)所占比例均显著高于IL-2组[(55.6±1.82)%vs(39.6±1.52)%,(16.6±1.82)%vs(9.8±1.30)%,(39.0±1.58)%vs(29.2±1.79)%;均P<0.05]。结论:自然沉降法从PBMC中分选T细胞具有低成本、高纯度、高回收率的优势。IL-7、IL-15和IL-21混合培养有利于早期记忆T细胞的生成,为肿瘤过继免疫治疗的体外细胞制备提供了实验依据。

Anti-viral memory T cell responses in the absence of IgG production in a COVID-19 convalescent individual

Cellular and humoral immunity are both important in host defense against severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).Although defects in SARS-CoV-2-specific T cell immunity have been found in patients with severe lung pathology,it is still largely unclear whether virus-specific T cells are sufficient for host protection.Here,we found that in a previously characterized cohort of convalescent subjects,one individual,though lacking detectable anti-viral neutralizing IgG antibodies,showed virus-specific T cell responses,both in CD4^(+)and CD8^(+)T cells.SARS-CoV-2-specific T cells in this and other individuals are maintained for up to 10 months.This study thus further supports a critical role of T cells in host defense against SARS-CoV-2,offering new insights into the design and evaluation of COVID-19 vaccines.

Distinct Pattern of Human Vδ/51 γδ/5 T Cells Recognizing MICA

γδ T cells represent one unique recognition pattern, the limited recognition, which distinguishes from the specific recognition for αβ T cells and pattern recognition for macrophages. Vδ1 γδT cell is the major subset of human γδT cells, which predominates in mucosal tissue including the intestinal epithelia. Presently, a few antigens that human Vδ1TCR can recognize have been identified. Among them, MHC class I chain-related molecules A （MICA） have been studied most intensively. Besides Vδ1TCR, MICA is also the ligand of NKG2D, a C-type lectin-like activating immunoreceptor. In human, only Vδ1 cells can simultaneously express both types of receptors of MICA while NK cells, αβ T cells and other subsets of γδT cells likewise express NKG2D. Although the precise mechanisms are still enigmatic, this distinct pattern of Vδ1 cells recognizing MICA predicts unique biological significance of Vδ1 cells in immune defense. Recent years, some progresses have been made in this issue. In this review we summarize the related reports and put forward some novel views based on our group＇s studies.

Vγ9Vδ2-T lymphocytes have impaired antiviral function in small-for-gestational-age and preterm neonates

Preterm and small-for-gestational-age （SGA） neonates are vulnerable groups that are susceptible to various microbial infections. Vγ9Vδ2-T cells are critical components of the host immune system and have been demonstrated to play an important role in the defense against viral infection in adults. However, the characteristics of Vγ9Vδ2-T cells in children, especially the preterm and SGA populations, are poorly understood. Here, we examined the frequency and antiviral function of Vγ9Vδ2-T cells in neonates, including preterm, SGA and full-term babies. When compared to adults, neonates had a significantly lower percentage of Vγ9Vδ2-T cells in the blood. Upon influenza virus stimulation, neonatalVγ9Vδ2-T cells, especially from preterm and SGA babies, showed markedly decreased and delayed antiviral cytokine responses than those of adults. In addition, the antiviral responses of neonatal Vγ9Vδ2-T cells were positively correlated with gestational age and birth weight. Finally, a weaker expansion ofVγ9Vδ2-T cells by isopentenyl pyrophosphate （IPP） was shown in neonates than the expansion in adults. Our data suggest that the depressed antiviral activity and decreased frequency of Vγ9Vδ2-T cells may likely account for the high susceptibility to microbial infection in neonates, particularly in preterm and SGA babies. Improving Vγ9Vδ2-T -cell function of neonates may provide a new way to defend against virus infection.

慢性乙型肝炎患者记忆性CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15Rβ表达和抗病毒治疗相关性研究

目的:探究慢性乙型肝炎患者外周血单个核细胞(PBMC)中记忆性CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15受体β亚基(IL-2/IL-15Rβ)表达及其与细胞增殖和分泌功能相关性,以及慢性活动性乙型肝炎(CAHB)患者治疗前后乙型肝炎病毒(HBV)感染相关指标、丙氨酸转氨酶(ALT)和CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15Rβ表达变化。方法:选取2019年3月至2020年12月皖南医学院附属第一医院感染科慢性乙型肝炎住院及门诊患者184例,根据纳入和排除标准,CAHB组纳入68例,无症状HBV携带组纳入47例。同时选取同时期30例健康体检者纳入健康对照组。CAHB组患者接受核苷(酸)类药物抗病毒治疗并随访(8例失访),分别比较剩余30例乙型肝炎e抗原(HBeAg)阳性和30例HBeAg阴性患者治疗前后HBV感染相关指标、ALT和CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15Rβ表达变化。多组间正态分布计量资料比较采用单因素方差分析;两组间正态分布计量资料比较采用配对样本t检验;两组间非正态分布计量资料比较采用Mann-Whitney U秩和检验。采用Pearson相关系数评估CAHB患者PBMC CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15Rβ表达比例和平均荧光强度(MFI)与CD3^(+)T淋巴细胞中CD8^(+)CD45RO^(+)T淋巴细胞比例相关性。P<0.05为差异有统计学意义。结果:CAHB组患者PBMC CD3^(+)T淋巴细胞中CD8^(+)CD45RO^(+)T淋巴细胞比例为(8.6±3.7)%,高于无症状HBV携带组[(5.7±2.5)%]和健康对照组[(5.5±1.5)%],差异均有统计学意义(P均<0.05);CAHB组PBMC CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15Rβ表达比例为(6.8±4.7)%,高于无症状HBV携带组[(4.7±2.8)%]和健康对照组[(4.3±2.2)%],差异均有统计学意义(P均<0.05);CAHB组PBMC CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15RβMFI为(243±168),高于无症状HBV携带组(160±91)和健康对照组(160±63),差异均有统计学意义(P均<0.05)。CAHB组PBMC CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15Rβ表达比例及MFI与CD3^(+)T淋巴细胞中CD8^(+)CD45RO^(+)T淋巴细胞比例均呈正相关(r=0.33和0.28,P均<0.05)。经抗-CD3+super-2刺激,CAHB组PBMC CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞增殖比例为(43.7±16.0)%,高于无症状HBV携带组[(29.1±9.4)%]和健康对照组[(26.8±9.6)%],差异均有统计学意义(P均<0.05);阻断IL-2/IL-15Rβ表达后,CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞增殖水平下降[(11.2±6.3)%],低于CAHB组,差异有统计学意义(P<0.05)。CAHB组CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞经抗-CD3+super-2刺激后分泌干扰素-γ(IFN-γ)、IL-2和肿瘤坏死因子-α(TNF-α)的CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞比例分别为(13.8±5.4)%、(14.0±4.3)%和(12.3±4.6)%,高于无症状HBV携带组[(8.4±2.6)%、(9.4±3.2)%和(6.8±3.3)%]和健康对照组[(6.9±2.7)%、(9.9±3.0)%和(7.7±3.8)%],差异均有统计学意义(P均<0.05);阻断IL-2/IL-15Rβ表达后,分泌IFN-γ[(2.4±1.6)%],IL-2[(4.1±1.9)%]和TNF-α[(4.1±1.8)%]的CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞比例均下降,低于CAHB组,差异均有统计学意义(P均<0.05)。30例HBeAg阳性CAHB患者治疗前HBeAg、ALT、PBMC CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15Rβ表达比例和MFI分别为521.4(68.9,1339.0)COI、292(160,528)U/L、(6.4±3.2)%和(239±136),高于治疗后水平[3.5(1.5,17.5)COI、20(14,31)U/L、(4.1±2.4)%和(134±58)],差异均有统计学意义(Z=5.337和6.403,t=3.229和3.892,P均<0.05)。30例HBeAg阴性CAHB患者抗病毒治疗前乙型肝炎表面抗原(HBsAg)、ALT、PBMC CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞IL-2/IL-15Rβ表达比例和MFI分别为(5310±2851)COI、(328±207)U/L、(7.1±5.8)%和(252±110),高于治疗后水平[(3811±2495)COI、(33±14)U/L、(4.6±2.9)%和(154±73)],差异均有统计学意义(t=2.167、5.595、2.116和2.383,P均<0.05)。结论:IL-2/IL-15Rβ表达水平与CAHB患者记忆性CD3^(+)CD8^(+)CD45RO^(+)T淋巴细胞数目、增殖和分泌功能密切相关。