地塞米松对系统性红斑狼疮患者外周血单个核细胞中细胞因子分泌及T细胞亚群表达的影响

目的 探讨地塞米松对系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMCs)中白细胞介素(IL)-17、干扰素(IFN)-γ分泌水平和Th17、Tc17、Th1、Tc1等T细胞亚群表达的影响.方法 SLE患者和健康对照者的PBMCs分空白孔、佛波酯(PMA)/离子霉素(Ionomycin)孔、PMA/Ionomycin+地塞米松(DEX)孔行体外培养,运用四色流式细胞术检测PBMCs表达Th17、Tc17、Th1、Tc1等T细胞亚群的百分比,采用酶联免疫吸附法测定SLE患者和健康对照者血浆及PBMCs培养上清中IL-17、IFN-γ的表达水平.结果 SLE组患者血浆IL-17[138.98(84.82～187.04)ng/L]、IFN-γ[21.92(15.95～27.09)ng/L]含量均高于正常对照组[57.21(47.78～72.12)ng/L,13.43(7.04～17.37)ng/L].无PMA刺激条件下,SLE患者PBMCs培养上清中细胞因子水平、PBMCs中各T细胞亚群的百分比与正常对照组相比差异均无统计学意义(P均＞0.05);加入PMA刺激后,SLE患者PBMCs上清中IL-17的水平[(26.43±10.04)ng/L]和外周血Th17[(2.49±1.49)%]、Tc1[(44.89±16.43)%]细胞的比例均显著高于正常对照组[(18.06±5.42)ng/L,(1.47±0.73)%,(31.41±9.05)%)(P均＜0.05),SLE患者Th1、Tc17细胞百分比与正常对照组相比,差异均无统计学意义(P均＞0.05);地塞米松能明显抑制活化状态下的PBMCs分泌IL-17的水平[(16.72±6.09)ng/L](P＜0.01),且显著下调Th17(1. 34±0.76)%、Tc1(34.62±17.25)%细胞百分比(P均＜0.05),而地塞米松对IL-17的抑制作用更强.结论 SLE患者体内T细胞亚群及其相应细胞因子的表达水平存在明显异常,地塞米松能干扰SLE患者体内细胞因子网络失衡的免疫病理过程,且对IL-17有明显的抑制作用,为临床使用糖皮质激素治疗SLE提供新的理论依据和实验室基础.     

免疫性血小板减少症患者外周血中滤泡辅助性T细胞和IL-21的水平及临床意义

目的探讨滤泡辅助性T细胞(Tfh细胞)和IL-21在免疫性血小板减少症(ITP)中的作用。方法采集15例ITP患者及20例健康对照者外周血,用流式细胞术检测外周血单个核细胞(PBMC)中CXCR5+CD4+Tfh细胞的百分率,ELISA法检测血清IL-21水平;用实时荧光定量PCR检测PBMC中IL-21 mRNA的表达。结果 ITP组PBMC中CXCR5+CD4+Tfh细胞比例(17.55±1.015)%显著高于健康人对照组(10.19±0.343)%,差异有统计学意义(t=7.657,P0.01);ITP组血清中IL-21水平(100.7±3.71)pg/mL显著高于健康人对照组(64.6±0.81)pg/mL,差异有统计学意义(t=10.84,P0.01),且两者之间存在明显相关性(r=0.689 3,P0.01)。ITP患者IL-21 mRNA的相对表达量4.319±0.767明显高于健康人对照组1.125±0.052,差异有统计学意义(t=4.813,P0.01)。结论 ITP患者外周血Tfh细胞比例、IL-21水平及IL-21 mRNA表达均增高,可能在ITP的发病机制中发挥重要作用。     

白细胞介素-7增强脓毒性心肌病患者辅助性T细胞9活性

目的观察白细胞介素-7(interleukin-7, IL-7)、辅助性T细胞9(T helper cell 9, Th9)、杀伤性T细胞9(cytotoxic T cell 9, Tc9)在脓毒性心肌病患者中的水平, 评估外源性IL-7对脓毒性心肌病患者Th9细胞的调控。方法采用横断面研究, 纳入2018年6月至2022年1月在郑州市第七人民医院就诊的脓毒性心肌病患者、脓毒症患者和对照者, 分离血浆和外周血单个核细胞(peripheral blood mononuclear cells, PBMCs), 重组IL-7刺激PBMCs, 酶联免疫吸附试验检测IL-7、IL-9、可溶型CD127水平, 流式细胞术检测T细胞中CD127表达和Th9、Tc9细胞比例, 实时定量PCR法检测富含嘌呤的核酸结合蛋白1(purine-rich nucleic acid binding protein 1, PU.1)和转录因子叉头框蛋白O1(forkhead box protein O1, Foxo1)mRNA表达。重组IL-7刺激脓毒性心肌病患者分选的Th9细胞后与自体CD8+T细胞、人脐静脉内皮...     

滤泡辅助性T细胞在急性肝衰竭小鼠模型中的变化及意义

目的:探讨滤泡辅助性T(T follicular helper,Tfh)细胞及相关细胞因子白细胞介素(interleukin,IL)-21在急性肝衰竭(acute liver failure,ALF)小鼠模型中的变化及意义。方法:将20只雌性C57BL/6小鼠随机分成2组,分别予脂多糖/D-氨基半乳糖或磷酸盐缓冲液(phosphate buffer solution,PBS)腹腔注射,6 h后采集血液、肝脏和脾脏组织标本。采用苏木精-伊红染色法分析肝脏病变情况;流式细胞术分析小鼠脾脏单个核细胞中Tfh细胞的频数;酶联免疫吸附试验分析血清中IL-21和肿瘤坏死因子α(tumor necrosis factorα,TNF-α)的水平。结果:ALF小鼠肝脏肿大而呈暗红色,肝细胞大量坏死和炎性细胞浸润,其血清中TNF-α及IL-21的水平均较正常对照组显著增高(P<0.01,P<0.05);而单个核细胞中CD4+CXCR5+、CD4+PD-1+、CD4+CXCR5+PD-1+T细胞频数亦较正常对照组显著升高(P<0.01)。结论:ALF小鼠脾脏单个核细胞中Tfh细胞频数显著增加,血清IL-21水平显著升高,推测Tfh细胞在ALF的发病过程中起重要作用,本研究结果可能为ALF的治疗提供新思路。     

传染性单核细胞增多症患儿外周血Th17细胞表达的研究

目的 探讨传染性单核细胞增多症(IM)患儿外周血Th17细胞、IL-17、IL-6水平的变化及临床意义.方法 分离IM患儿和健康者外周血单个核细胞(PBMC),免疫磁珠阴选CD4+T淋巴细胞,然后加佛波酯/离子霉素(PMA/Ion),经固定,透膜处理进行细胞内染色,流式细胞术检测CD4+T淋巴细胞内IL-17、IL-6水平.结果 IM患儿急性期IL-17、IL-6表达水平显著高于恢复期(P＜0.01),两者又显著高于健康对照组(P＜0.01).结论 IM患儿外周血中存在异常增高的Th17细胞及其分泌的细胞因子,且其水平与病情活动相关,是判断IM患儿免疫状态和预后的指标之一.     

乙型肝炎患者e抗原转换与外周血γδT和Tc17的关系

目的探讨乙型病毒性肝炎患者e抗原(HBeAg)转换与外周血γδT、白细胞介素(IL)-17、γ干扰素(IFN-γ)等的相关性及临床意义。方法随机选取2017年10月至2018年4月在该院就诊的乙型肝炎患者90例作为研究组,选取同期30例健康人作为对照组。然后在研究组内根据HBeAg表达与否进一步分为3个亚组:HBeAg+/Ab-组(n=30)、HBeAg+/Ab+组(n=30)、HBeAg-/Ab+组(n=30)。运用流式细胞仪检测受试者外周血中的γδT细胞、IL-17+γδT细胞、IFN-γ+γδT细胞、IL-17+CD8细胞在外周血单个核细胞(PBMCs)中的比例,对比上述指标在研究组及对照组间、以及乙型肝炎患者HBeAg/Ab不同表达组内的差异。进一步分析乙型肝炎患者HBeAg/Ab不同表达组及不同血清丙氨酸氨基转移酶(ALT)水平下,血清内HBV-DNA水平的差异。结果研究组IFN-γ+γδT细胞、γδT细胞水平、IL-17+CD8细胞要明显高于对照组(P<0.05)。研究组中,HBeAg+/Ab-组和HBeAg+/Ab+组患者PBMCs内总γδT细胞水平要显著高于HBeAg-/Ab+组(P<0.05);HBeAg+/Ab+组乙型肝炎患者IL-17+CD8细胞明显高于HBeAg+/Ab-组和HBeAg-/Ab b组(P<0.05)。HBeAg+/Ab-组慢性乙型肝炎患者血清内HBV-DNA水平要明显高于HBeAg+/Ab+组和HBeAg-/Ab+组(P<0.05)。结论γδT细胞在HBeAg/Ab血清转换中可能发挥一定作用。IL-17+CD8细胞在慢性乙型肝炎良性转归中起重要作用,对于抗病毒治疗的效果可能具有预测意义。     

强直性脊柱炎外周血Th17细胞的检测及意义

目的探讨Th17细胞在强直性脊柱炎(AS)患者外周血中的水平及意义。方法取20例AS患者(分AS病情稳定组及活动组各10例)和健康人外周血单个核细胞(PBMC)、免疫磁珠分选CD4+T细胞,用或不用非特异性刺激剂(anti-CD3、anti-CD28),然后加佛被酯/离子霉素,经固定/透膜处理进行细胞内染色,流式细胞术检测CD4+T细胞内白细胞介素-17(IL-17)、γ-干扰素(IFN-γ)、及IL-6水平。结果免疫磁珠分选CD4+T细胞纯度达90%以上。AS病情活动组IL-17表达水平较病情稳定组和健康对照组显著增高,差异有统计学意义(P<0.01)。用anti-CD3、anti-CD28刺激后,CD4+T细胞IL-17胞内表达水平较无刺激有一定的增加。AS患者CD4+T细胞IFN-γ胞内表达水平呈现与IL-17表达相似的特点。结论 AS患者外周血CD4+T细胞胞内IL-17和IFN-γ呈高表达,提示分泌IL-17的Th17细胞和分泌IFN-γ的Th1细胞共同参与了AS发病过程。     

系统性红斑狼疮患者外周血淋巴细胞CD28/CTLA-4分子表达的研究

目的探讨系统性红斑狼疮(systemic lupus erythematosus,SLE)患者外周血淋巴细胞CD28/CTLA-4分子和CD28/CTLA-4mRNA的表达。方法研究对象为SLE患者49例(活跃期30例、缓解期19例)及对照组23例。外周血单个核细胞(peripheral blood mononuclear cell,PBMCs)经梯度密度离心法分离后分佛波醇乙酯(PMA)(10ng/ml)及伊屋诺霉素(500ng/ml)刺激组和不加刺激剂组两组培养。将PBMCs分别培养24、48、72及96小时。应用流式细胞计量术(flow cytometry,FCM)检测外周血淋巴细胞培养前后CD28及CTLA-4分子的表达。采用RT-PCR方法检测CD28mRNA和CTLA-4mRNA的表达。结果刺激前后SLE患者CD3+及CD8+T细胞上CD28分子表达量与对照组比较差异均无统计学意义(P>0.05)。刺激前活跃期SLE患者CD3+T细胞上CTLA-4分子表达量较对照组明显降低[(0.78±0.51)%vs(1.34±0.76)%,P<0.05]。刺激后72小时SLE患者CD3+T细胞及CD8+T细胞上CTLA-4分子表达量仍低于对照组,但差异无统计学意义(P>0.05)。刺激前后PBMCs中CD28mRNA及CTLA-4mRNA表达情况与刺激前后CD3+T细胞上CD28及CTLA-4分子变化情况相似。刺激前SLE患者CD3+T细胞上CTLA-4分子表达量与SLE活动指数(SLEDAI)呈显著的直线负相关关系(P<0.001)。结论 SLE患者T细胞CTLA-4分子存在表达及上调机制障碍,提示CD28分子及CTLA-4分子存在功能失衡,这种失衡可能通过一定的机制参与SLE的发病过程。     

滤泡辅助性T细胞在类风湿关节炎发病中的临床意义

目的通过检测类风湿关节炎(RA)患者外周血滤泡辅助性T淋巴细胞(Tfh))和白细胞介素-21(IL-21)等的表达,探讨其在RA发病过程中可能的免疫学发病机制。方法根据病情活动度不同将病例组分为轻、中、重度活动组(各30例),健康对照组30例;流式细胞仪检测RA和健康对照组外周血单个核细胞(PBMCs)中CD4-异硫氰酸荧光素(FITC)、CXCR5-藻红蛋白(PE)、ICOS-别藻蓝蛋白(APC)标记的CD3+CD4+CXCR+、CD3+CD4+ICOS+及CD3+CD4+CXCR+ICOS+Tfh百分率;酶联免疫吸附试验(ELISA)检测IL-21、类风湿因子(RF-IgM)、抗环瓜氨酸肽(CCP)抗体(CCP-IgG)及红细胞沉降率(ESR)、C反应蛋白(CRP),分析其与Tfh的相关性。结果 RA轻、中、重度活动组患者外周血中Tfh百分率明显高于健康对照组(P0.05);Tfh百分率随着RA活动度的增加呈上升趋势,组间差异有统计学意义(P0.05);Tfh细胞百分率与RA患者ESR、CRP、RF及抗CCP抗体滴度呈正相关,差异有统计学意义(P0.05)。结论 RA患者外周血Tfh、IL-21、CRP、ESR、RF及CCP水平明显升高,与疾病活动度等有明显相关性,提示Tfh细胞可能在RA发病中起着一定的作用。     

不同刺激剂对人外周血淋巴细胞亚群增殖的影响

目的:采用流式细胞术分析PHA、PMA、IL-2对PBMC和全血不同培养体系中人外周血淋巴细胞亚群增殖的影响,为不同的实验目的选取合适的培养体系提供实验基础。方法:收集健康受试者(n=6)静脉血肝素钠抗凝样本(10 ml/样本),每个样本取300μl全血并直接裂解红细胞经荧光染色后流式细胞术检测淋巴细胞亚群。在400μl全血接种淋巴细胞自体血浆培养液中,分别加入3种不同刺激剂,37℃、5%CO2培养箱中培养60 h;2 ml全血分离PBMC,淋巴细胞培养液中分别加入3种不同刺激剂,于37℃、5%CO2培养箱中培养60 h。培养物经荧光染色后流式细胞术分析淋巴细胞亚群变化。结果:分离的PBMC经PHA刺激后淋巴细胞主要表达CD4-CD8-CD3+淋巴母细胞;PMA刺激后CD3+CD4+T淋巴细胞和CD3-CD19+B淋巴细胞比例下降(P 0.01, P0.05);IL-2刺激后淋巴细胞亚群比例增殖前后无明显变化。全血自体血浆经PHA刺激后转化为CD4+CD3+T淋巴母细胞和CD8+CD3+T淋巴母细胞,B淋巴细胞和NK细胞未见表达;PMA刺激后转化为CD8+CD3+T淋巴母细胞和CD4-CD8-T淋巴细胞,B/NK细胞用表面标志物无法区分;IL-2刺激后NK细胞比例明显增加(P0.05)。结论:PMA刺激增殖作用最快,IL-2对淋巴细胞亚群增殖前后比例影响最小,PHA刺激淋巴细胞分裂代数更多。     

Decreased IL-7 signaling in T cells from patients with PTLD after allogeneic HSCT

Interleukin (IL)-7, a nonredundant cytokine for B and T cells, plays a central role in cell survival and immune memory formation. Peripheral blood mononuclear cells (PBMCs) from 7 patients after hematopoietic stem cell transplantation (HSCT) diagnosed with posttransplantation lymphoproliferative disease (PTLD) and from 10 Epstein-Barr virus (EBV) polymerase chain reaction-positive HSCT patients (controls) were evaluated for IL-7- and IL-2 induced Stat5 phosphorylation in CD4+ and CD8+ T cells. PBMCs from PTLD+ and control patients exhibited detectable EBV specific CD8+ T cells defined by tetramer analysis. CD4+ and CD8+ T cells from patients with PTLD showed statistically significant reduction in responsiveness to IL-7 compared with PBMCs obtained from controls defined by Stat5 phosphorylation. CD20+ B cells from patients with PTLD and from some EBV+ polymerase chain reaction control individuals exhibited IL-7R expression. Dysregulated immune surveillance, reflected by deficient Stat5 phosphorylation, may facilitate PTLD development despite the presence of EBV-reactive CD8+ T cells. Reduced IL-7 responsiveness will aid to monitor patients after HSCT for increased risk to develop EBV-associated PTLD.     

手足口病患儿外周血单个核细胞中TLR4通路检测与病情、炎症反应、免疫功能的关系

目的研究手足口病(HFMD)患儿外周血单个核细胞(PBMCs)中Toll样受体4(TLR4)通路与病情、炎症反应、免疫功能的关系。方法回顾性选择2016年7月至2019年4月期间在中国医科大学附属第一医院诊断为HFMD的患儿并根据病情分为轻症组和重症组,另取同期体检的健康儿童作为对照组。测定各组对象PBMCs中TLR4、核因子κB(NF-κB)的表达水平,血清中炎症细胞因子、免疫球蛋白的含量,外周血中免疫细胞的含量。分析HFMD患儿TLR4的表达水平与血清C反应蛋白(CRP)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、IgM、IgG、IgA、CD3^+CD4^+T细胞、CD3^+CD8^+T细胞、CD19^+B细胞的相关性。结果重症组和轻症组患儿PBMCs中TLR4、NF-κB的mRNA相对表达水平及血清中C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、免疫球蛋白M(IgM)的含量均明显高于对照组血清中IgG、IgA的含量及外周血中CD3^+CD4^+T细胞、CD3^+CD^8^+T细胞、CD19^+B细胞的含量均明显低于对照组(P <0.05),且重症组患者上述指标的变化较轻症组更为显著(P <0.05);TLR4与CRP、TNF-α、IL-6、IgM呈正相关,与IgG、IgA、CD3^+CD4^+T细胞、CD3^+CD8^+T细胞、CD19^+B细胞呈负相关。结论手足口病患儿PBMCs中TLR4通路的过度激活与病情加重、炎症反应激活、免疫功能紊乱有关。     

Cytokine interactions in human immunodeficiency virus-infected individuals: roles of interleukin (IL)-2, IL-12, and IL-15

Cytokines have been shown to be powerful regulators of the immune response. In this study, we analyze the effect that the newly recognized cytokine interleukin (IL)-15 has on proliferation and cytokine induction using peripheral blood mononuclear cells (PBMCs) and purified CD4+ T cells from patients infected with human immunodeficiency virus (HIV) who are at various stages in their disease. We observed that IL-15 enhances the proliferative response in a dose-dependent manner from PBMCs of HIV-infected individuals when stimulated by polyclonal mitogen, tetanus toxoid, or HIV-specific antigen. The effects of exogenous IL-15 are substantially diminished by adding a neutralizing antibody to the beta chain of the IL-2 receptor. Moreover, the ability of IL-15 to increase proliferation is enhanced by the presence of endogenous IL-2 produced in the cultures. The effect that exogenous IL-15 had on IL-2, IL-4, and interferon (IFN)-gamma induction from PBMC's or CD4+ T cells in response to mitogen or tetanus toxoid was also examined. This was compared to the effect that exogenous IL-2 and IL-12 had under the same conditions. Addition of IL- 2 or IL-15 to short-term in vitro cultures of either PBMCs or CD4+ T cells had little effect on IL-2, IL-4, or IFN-gamma production. By contrast, IL-12 caused substantial enhancement of both IL-2 and IFN- gamma production from these cultures. The role that endogenous cytokines have on IFN-gamma induction was also studied. Addition of a neutralizing antibody to the alpha chain of the IL-2 receptor or IL-12 to antigen stimulated cultures caused a striking decrease in IFN-gamma production. Neutralization of endogenous IL-15 also resulted in diminished IFN-gamma production from cultures stimulated with mitogen. IL-4 and IFN-gamma protein production by PBMCs and CD4+ T cells stimulated with mitogen was assessed to see if we could detect a specific bias of cytokine production. Small amounts of IL-4 were detected from CD4+ T cells but not PBMCs from most individuals tested. IFN-gamma and IL-2, however, were also produced from these same cultures. These results further elucidate the mechanism of cytokine regulation in HIV-infected individuals, and they provide evidence that IL-15 may be a useful immune modulator.     

Recombinant interferon-alpha selectively inhibits the production of interleukin-5 by human CD4+ T cells.

The effects of recombinant IFN-alpha on the production of IL-5 by human CD4+ T cells were first analyzed on resting CD4+ T cells purified from normal PBMC and stimulated either with a combination of PMA and anti-CD28 mAb or anti-CD3 mAb cross-linked on B7-1/CD32-transfected mouse fibroblasts. We found that IFN-alpha profoundly inhibited in a dose-dependent manner IL-5 production by resting CD4+ T cells whereas IL-10 was upregulated in both systems. The addition of a neutralizing anti-IL-10 mAb to PMA and anti-CD28 mAb upregulated IL-5 production by resting CD4+ T cells but did not prevent IFN-alpha-induced IL-5 inhibition. We then analyzed the effect of IFN-alpha on the production of cytokines by differentiated type 2 helper (Th2) CD4+CD3- cells isolated from peripheral blood of two patients with the hypereosinophilic syndrome. In both cases, IFN-alpha markedly inhibited IL-5 production while it induced mild upregulation of IL-4 and IL-10. Finally, the inhibitory effect of IFN-alpha on IL-5 production was confirmed on a panel of Th2 and Th0 clones generated in vitro. In 2 out of 6 clones, IL-5 inhibition was associated with upregulation of IL-4 and IL-10. We conclude that IFN-alpha selectively downregulates IL-5 synthesis by human CD4+ T cells.     

Interleukin 2 induction of pore-forming protein gene expression in human peripheral blood CD8+ T cells

Our studies have analyzed pore-forming protein (PFP) mRNA expression in resting and stimulated human peripheral blood CD3- large granular lymphocytes (LGL), CD3+ T cells, and their CD4+ or CD8+ subsets. Signals that stimulate T cells to develop cytotoxic activity (i.e., IL- 2 or OKT-3 mAb) led to the induction of PFP mRNA in T cells. The data indicated that IL-2 directly increased PFP mRNA in the CD8+ subset of T cells, in the absence of new DNA or protein synthesis. Abrogation of IL- 2-induced PFP mRNA expression and cytotoxic potential of T cells by the anti-p75 IL-2 receptor mAb suggested that low numbers of p75 IL-2 receptors on CD8+ T cells were capable of transducing signals responsible for these IL-2-induced effects. The induction of T cell PFP mRNA via CD3, using OKT-3 mAb, was less rapid but greater than that caused by IL-2; however, a combination of PMA and ionomycin, which bypasses crosslinking of the TCR/CD3 complex, could not mimic this increase in PFP mRNA levels in T cells. The role of second messenger systems in regulating PFP mRNA expression remains to be determined. In contrast, high constitutive PFP mRNA expression was observed in CD3- LGL and these mRNA levels could not be enhanced by stimulation with IL- 2. The cytotoxic potential of peripheral blood T cells and LGL induced in response to IL-2 correlated with IL-2-induced PFP mRNA levels in these cells and was consistent with PFP being one of several important molecules involved in the effector function of cytotoxic lymphocytes.     

中国HCV/HIV合并感染者T淋巴细胞表面CD81、CXCR3表达的研究

目的探讨T淋巴细胞表面受体CD81、CXCR3，在丙肝病毒（HCV）单纯感染，艾滋病病毒（HIV）单纯感染和HCV／HIV合并感染过程中的表达及意义。方法采用流式细胞术，检测HCV感染组（n=21），HIV感染组（n=14），HCV／HIV感染组（n=28）及正常对照组（n=30）人外周血CD4^＋T淋巴细胞和CD8^＋T淋巴细胞表面CD81、CXCR3的表达。结果HIV感染组及HCV／HIV合并感染组CD4^＋T细胞表面CD81、CXCR3表达显著降低（P〈0．001），CD8^＋T细胞表面CD81、CXCR3表达显著升高（P〈0．001）；HCV感染组CD4^＋及CD8^＋T细胞表面CXCR3表达轻度升高但差异不显著，CD81在CD4^＋T细胞表面轻度升高而在CD8^＋T细胞表面明显升高（P〈0．01）。结论中国HCV／HIV合并感染患者外周血T淋巴细胞表面受体CD81、CXCR3，在CD4^＋T细胞表面表达降低，而在CD8^＋T细胞表面表达升高。     

Does cyclosporin A affect CCR5 and CXCR4 expression in primary HIV-1-infected patients?

BACKGROUND: CCR5 and CXCR4 are the major coreceptors of HIV required for successful viral entry. No information exists on the effect of cyclosporin A (CsA) on expression of CCR5 and CXCR4. A longitudinal study of the coreceptors' expression in freshly isolated peripheral blood mononuclear cells (PBMC) of patients with primary HIV infection (PHI) was performed. METHODS: Patients received highly active antiretroviral therapy (HAART) alone (n = 7) or with CsA (HAART + CsA) (n = 8). Flow cytometric data were analyzed at T0 (baseline), two (T2), six (T6), and twelve (T12) months after therapy initiation. RESULTS: At T0 PHI subjects presented a statistically significant higher count and percentage of CD8+CCR5+ lymphocytes compared to healthy donors (HD) (mean +/- SD, 2,240 +/- 1,998 vs 181 +/- 89 cells/microl). Conversely, CD4+CXCR4+ lymphocytes were less abundant in PHI than in HD (443 +/- 337 vs 673 +/- 339 cells/microl), whereas CD4+CCR5+ lymphocytes were substantially comparable (169 +/- 167 vs 126 +/- 60 cells/microl). In the follow up no differences between HAART and HAART + CsA groups reached statistical significance in CD4 lymphocytes. CD4+CCR5- lymphocytes displayed a rapid recovery after therapy initiation, similarly to the CD4+CXCR4+ subset. In CD8 lymphocytes a statistically significant difference between HAART and HAART + CsA patients occurred at T2 when HAART + CsA patients presented a lower absolute count of the CD8+CXCR4+ subset compared to the HAART group. The major change after therapy initiation in all PHI patients was a striking drop of CD8+CCR5+ lymphocytes; moreover, the CD8+CXCR4- subset behaved similarly. The decrement of CD8+CCR5+ lymphocytes paralleled the decline of viremia and CD8+CD38+ lymphocytes, with the sharpest slope at T2. Conversely, RANTES levels increased at T2 and remained elevated during the follow up. CONCLUSIONS: CsA cotreatment in PHI patients appears not to substantially modify HIV coreceptors' expression in PBMC. However, this novel piece of information should be used with caution, since this was not a randomized study between the HAART and the HAART + CsA groups.