The immunomodulatory effects of regulatory T cells: implications for immune regulation in the skin

Regulatory T cells are thought to have a critical role in the suppression of immune responses. In addition to the prevention of the development of autoimmunity, they are also thought to have a role in the prevention of allergic responses to environmental allergens, immune responses to tumours and the development of memory responses to chronic infections. They have been isolated within the skin and have been shown to express surface markers that enable skin-specific migration, suggesting that regulatory T cells have a functional role in the skin. There is accumulating evidence to suggest that regulatory T cells may be involved in numerous skin disorders and may also be modified by various therapeutic agents used to treat these disorders. We review the evidence for the presence of this T-cell subset in humans, the suppressive effects of regulatory T cells, and their role in the skin.     

慢性荨麻疹患者外周血中CD8~+CD28~-T细胞检测结果分析

目的:探讨CD8~+CD28~-T细胞在慢性荨麻疹患者致病机制中的作用。方法:收集临床慢性荨麻疹(CU)病例83例,同时设立对照组64例进行比较。应用ELISA法检测研究对象血清中抗Ig E抗体、抗FcεRⅠ抗体浓度,流式细胞仪检测外周血中CD3~+、CD4~+、CD8~+、CD8~+CD28~-、CD4~+CD25~+T细胞比例,分析检测结果。结果:83名CU中,23例抗Ig E抗体为阳性,占27.7%(27/83);31例抗FcεRⅠ抗体为阳性,占37.3%(31/83)。CU患者外周血CD8~+T细胞比例、CD8~+CD28~-T细胞比例低于正常对照组(P0.05),CD4~+/CD8~+比值、CD4~+CD25~+T细胞比例均高于对照组(P0.05)。结论:CU患者机体外周血CD8~+CD28~-T细胞、CD4~+CD25~+T细胞比例与对照组相比存在差异,CD8~+CD28~-T细胞比例降低也可能是CU致病机制之一。     

带状疱疹患者CD_4~+T淋巴细胞亚群的检测

目的通过观察带状疱疹患者T细胞亚群的变化,分析机体免疫功能与患带状疱疹的相关性,揭示带状疱疹发病的规律和机理。方法采用流式细胞仪技术检测73例不同年龄和性别的带状疱疹患者T细胞亚群的分布情况,并选择88例健康体检者做为健康对照。结果带状疱疹患者血清CD4+T细胞显著降低,CD3+T细胞降低,CD8+T细胞升高,CD4/CD8比值有所下降;不同年龄和不同病程的患者间也存在T细胞亚群的明显差异。结论带状疱疹患者存在CD4+T细胞的降低和T细胞亚群免疫功能下降,其在带状疱疹的发生和发展中可能起重要作用;老年人是带状疱疹发病的危险人群。     

Imbalance of CD4+CD29+ and CD4+CD45RA+ T-helper cell subsets in peripheral blood of patients with severe atopic dermatitis

To determine the proportion of T-helper cell subsets in the peripheral blood we studied 16 patients with mild, moderate and severe atopic dermatitis. Lymphocytes were isolated from heparinized peripheral blood and analysed by two-colour flow cytometry. Patients with severe atopic dermatitis had a decreased CD4+CD29+CD4+CD45RA+ ratio (p<0.01). We found a decreased absolute number of CD4+CD29+ cells (p<0.05) and an increased absolute number of CD4+CD45RA+ cells (p<0.05) in the peripheral blood. No significant changes in the CD4+CD29+CD4+CD45RA+ ratio were found in the peripheral blood of patients with clinically mild or moderate atopic dermatitis.     

神经梅毒合并HIV阳性患者的脑脊液及血液指标分析

目的：探讨脑脊液实验室检测在神经梅毒合并HIV阳性患者中的诊断价值及血清快速血浆反应素试验（rapid plasma reagent test, RPR）滴度、CD4+T细胞计数在神经梅毒腰穿指征中的应用。方法：收集2015年1月至2019年12月就诊于北京佑安医院的梅毒合并HIV阳性患者106例,采集脑脊液(cerebrospinal fluid, CSF)进行脑脊液白细胞（CSF-WBC）、脑脊液蛋白（CSF-protein）及脑脊液梅毒螺旋体颗粒凝集试验(treponema pallidum particle assay, TPPA)、RPR滴度检测,采集血液进行RPR滴度、CD4+T细胞计数检测,根据神经梅毒的诊断分神经梅毒组和非神经梅毒组,对两组的脑脊液检测结果、血清RPR滴度及CD4+T细胞计数检测结果进行分析。结果：106例梅毒合并HIV阳性患者中神经梅毒发病率为33.02%,CSF-RPR及CSF-TPPA对HIV阳性梅毒患者发生神经梅毒的诊断敏感性为68.57％和97.14％,特异性为92.96％和49.29％;CSF-WBC和CSF-protein的ROC（受试者工作特征曲线）分析曲线下面积（area under curve, AUC）分别为0.911和0.913,CSF-WBC为10.5/μL、CSF-protein为272.15 mg/L时,约登指数最大;血清RPR≥1∶16患者发生神经梅毒的几率是血清RPR<1∶16患者的1.52倍（OR 1.52,CI 1.14～2.04,P<0.05）,CD4+T细胞≤350个/μL发生神经梅毒的几率为CD4+ T细胞>350个/μL患者的2.37倍（OR 2.37, 95％ CI 1.64～3.41,P<0.05）。结论：HIV阳性患者神经梅毒的发病率较高,CSF-RPR对HIV阳性患者发生神经梅毒具有较高的诊断价值,血清RPR滴度≥1∶16和CD4+T≤350个/μL,是HIV阳性患者神经梅毒的危险因素。     

Allergic contact dermatitis: the cellular effectors

Contact hypersensitivity reactions are mediated by lymphocytic effector cells. Until recently it was believed that the most important of these were CD4+ T lymphocytes. However, there is growing evidence that in many instances the predominant effector cell may be a CD8+ T lymphocyte, with in some instances CD4+ cells performing a counter-regulatory function. Here we review the roles of CD4+ T helper (Th) cells and CD8+ T cytotoxic (Tc) cells, and their main functional subpopulations (respectively, Th1 and Th2 cells and Tc1 and Tc2 cells) in the elicitation of contact hypersensitivity reactions and consider the implications of effector cell selectivity for the biology of allergic contact dermatitis.     

Phagocytized apoptotic cells in subcutaneous panniculitis-like T-cell lymphoma

We reported on a case of subcutaneous panniculitis-like T-cell lymphoma (SPTCL) with multiple erythematous nodular lesions on the extremities, trunk and face. Histological examination of an excised lesion revealed a dense infiltrate of markedly atypical T-lymphoid cells expressing the CD8+ phenotype located in the subcutaneous tissue with histiocyte-phagocytizing apoptotic cells. The 'bean-bag' histiocytic cells, the characteristic finding of SPTCL, are considered to be products of haemophagocytosis. In our case the 'bean-bag' cells were produced by phagocytosis of apoptotic bodies, as confirmed by electron microscopy. It is suspected that 'bean-bag' cells are related not to haemophagocytosis but to phagocytosis of apoptotic cells in the CD8+ T-cell type of SPTCL.     

CD4~+ CD25~+调节性T细胞与IL-17在梅毒血清固定患者外周血中的表达及意义

目的:观察梅毒血清固定患者外周血中CD4+CD25+调节性T细胞和IL-17的表达及意义。方法:采用三色流式细胞术及免疫酶联吸附法测定18例梅毒血清固定患者和18例对照者外周血中CD4+CD25+调节性T细胞和IL-17的表达水平。结果:梅毒血清固定患者外周血CD4+CD25+调节性T细胞的含量明显高于对照组(t=6.29,P<0.01);IL-17的含量与对照组比较差异无统计学意义(t=-1.68,P=0.102)。结论:CD4+CD25+调节性T细胞表达水平增高造成的免疫抑制可能与梅毒血清固定有关;IL-17的表达水平可能与梅毒血清固定无关。     

Immunoregulation of allergic contact dermatitis

Allergic contact dermatitis (ACD) to haptens can serve as a valuable paradigm for understanding the physiopathology of T cell mediated immune responses. In sensitized individuals, exposure to the relevant hapten initiates clinical expression of ACD, which depends on the rapid activation of specific T cells. Mechanisms of tissue damage include direct cytotoxicity against keratinocytes, mostly mediated by CD8+ T cells, and T cell release of cytokines, which amplify the inflammatory response by targeting resident skin cells. The expression of ACD is actively regulated by specialized subsets of T lymphocytes with suppressive functions. In particular, T regulatory cells producing high levels of IL-10 suppress ACD by blocking the functions of dendritic cells. In contrast CD4+CD25+ regulatory T cells prevent immunopathological reactions and maintain peripheral tolerance to haptens by acting via a cell-to-cell contact mechanism. Understanding the role of suppressor T cells and the requirements for their in vivo and in vitro expansion are critical steps for the development of specific desensitization protocols in hapten-allergic individuals. This information may also provide the basis for novel interventions in other immune-mediated diseases.     

CD4~+CD25~+调节性T细胞及其细胞因子在寻常型银屑病患者的改变

目的探讨寻常型银屑病患者CD4~+CD25~+调节性T细胞及其细胞因子的改变。方法用流氏细胞仪直接免疫荧光法检测寻常型银屑病患者外周血CD4~+CD25~+T细胞的百分率、FoxP3+分子表达、细胞因子白细胞介素(IL)-10、转化生长因子(TGF)-β的表达。结果寻常型银屑病患者CD4~+CD25~+T细胞在进行期组、静止期组、健康对照组结果分别是4.55±1.11、5.26±1.38、6.03±1.96,CD4~+CD25~+FoxP3+T细胞在进行期组、静止期组、健康对照组结果分别是3.85±0.89、4.31±1.26、4.95±1.37。进行期患者明显低于静止期患者和健康对照组(P0.05);而静止期患者与健康对照组比较差异无统计学意义(P0.05)。CD4~+CD25~+IL-10+细胞、CD4~+CD25~+TGF-β+T细胞在进行期组、静止期组、健康对照组结果分别是11.57±5.64、14.49±8.26、16.35±9.98;2.65±0.97、4.23±1.55、4.83±1.84;进行期患者明显低于静止期患者和健康对照组(P0.05);而静止期患者与健康对照组比较差异无统计学意义(P0.05)。CD4~+CD25~+FoxP3+T细胞与PASI呈负相关(r=-0.659,P0.05);CD4~+CD25~+IL-10+T与PASI呈负相关(r=-0.537,P0.05);而CD4~+CD25~+TGF-β+T细胞与PASI直线相关(r=-0.184,P0.05)。结论 CD4~+CD25~+调节性T细胞的比例以及Foxp3的表达、IL-10、TGF-β的表达明显下降,导致调节性T细胞的抑制功能缺陷,进而影响银屑病病情的发展。可以推测,调节性T细胞的数量或功能异常,均将影响银屑病的发病。     

应用流式细胞仪研究深圳地区健康成人及HIV/AIDS患者的免疫状况

目的：研究深圳地区健康人、HIV无症状感染者及AIDS患者的免疫状态，以了解艾滋病早期T细胞亚群的变化过程。方法：应用流式细胞仪三色荧光单克隆抗体（CD4－FITC／CD8－PE／CD3－PC5）检测34例正常健康成人，12例HIV感染无症状者，23例AIDS患者的外周血CD4^ 、CD8 ^ T淋巴细胞计数及CD4^ /CD8^ 比值。结果：HIV感染无症状者及AIDS患者的CD4^ 淋巴细胞及CD4^ /CD8比值均明显低于健康成人（P＜0．01）；HIV感染无症状者CD8^ T淋巴细胞明显高于正常健康人和AIDS患者（P＜0．01）；CD4^ T淋巴细胞随病程进展不断下降，AIDS患者与HIV感染无症状者之间存在着显著差异。结论：T淋巴细胞免疫状况可作为评价AIDS病程进展程度的重要指标。     

Lymphomatoid papulosis subtype C: A case report and literature review

Lymphomatoid papulosis (LyP) is a rare CD30⁺ lymphoproliferative primary skin disease with a benign clinical course and malignant histopathology. LyP is classified into seven subtypes based on histopathology: subtypes A through F and LyP with 6p25.3 chromosome rearrangement. We present here, a case report of a 51‐year‐old man, afflicted with multiple papules and nodules on his left arm for over 3 months and diagnosed with LyP subtype C. The patient refused treatment, and his lesions faded with no visible rash on the left arm 14 months after diagnosis.     

系统性硬皮病CD4~+ T细胞中CD70 mRNA及蛋白表达水平的观察

目的:研究系统性硬皮病(SSc)患者外周血CD4+ T细胞中CD70的表达水平。方法:应用密度梯度离心法分离17例SSc患者(女性12例,男性5例)和13例对照者(女性8例,男性5例)的外周血单个核细胞,磁珠分选CD4+ T细胞。RT-PCR检测CD4+ T细胞中CD70 mRNA的表达水平;流式细胞术检测CD70蛋白在CD4+ T细胞的表达水平。结果:与对照组相比,SSc患者CD4+ T细胞中CD70mRNA和CD70蛋白表达均明显升高(P=0.001;P=0.007)。结论:CD70在SSc的发生发展中可能起着重要作用。     

系统性红斑狼疮患者CD4~+ CD25~+调节性T细胞及其细胞因子的改变

目的探讨系统性红斑狼疮(SLE)患者CD4+CD25+调节性T细胞(Tr)及其细胞因子的改变。方法用流式细胞仪直接免疫荧光法检测SLE患者外周血CD4+CD25+T细胞的百分率,FoxP3+分子表达,细胞因子IL-10,TGF-β1的表达。结果 SLE患者活动期组、稳定期组、健康对照组CD4+CD25+T细胞占CD4+T细胞的百分率分别是(3.57±1.45)%,(5.14±1.63)%,(6.03±1.96)%。活动期患者明显低于稳定期患者和健康对照组,差异均有统计学意义(P均<0.05)。CD4+CD25+FoxP3+T细胞、CD4+CD25-FoxP3+T细胞占CD4+T细胞的百分率,活动期患者明显高于稳定期患者和健康对照组,差异均有统计学意义(P<0.05)。CD4+CD25+FoxP3+T细胞与疾病的活动指数(DAI)正相关(r=0.659,P<0.05);而CD4+CD25-FoxP3+T细胞与DAI无统计学相关性(r=0.184,P>0.05)。活动期患者CD4+CD25+IL-10+细胞明显高于稳定期患者和健康对照组,差异均有统计学意义(P均<0.05)。然而CD4+CD25-TGF-β+细胞在活动期患者、稳定期患者和健康对照组之间比较差异无统计学意义(P均>0.05)。结论 SLE患者外周血中Tr细胞数量减少,分泌的IL-10增加,TGF-β减少,可能导致细胞免疫抑制功能减弱,自身免疫耐受平衡被打破,出现激活的自身反应性T细胞增多;此外,T淋巴细胞激活增多,还可辅助B细胞产生更多相关抗体,表现出多种临床症状。     

Regulation of nickel-induced T-cell responsiveness by CD4+CD25+ cells in contact allergic patients and healthy individuals

In this study, we investigated the capacity of CD4⁺CD25⁺ regulatory T cells to suppress nickel‐specific effector T cells, both in nickel‐allergic patients and healthy controls. CD4⁺ cells isolated from allergic patients showed an increased proliferative response to nickel, whereas CD4⁺ cells from negative controls did not respond to allergen. When CD4⁺CD25⁺ cells were depleted, nickel‐specific responsiveness was strongly increased both in allergic and in non‐allergic individuals, with the most pronounced effect in allergic patients. These regulatory T cells were anergic to nickel but inhibited nickel‐specific CD4⁺CD25^– effector T cells in coculture experiments. CD4⁺CD25⁺ cells from nickel‐allergic patients showed only a limited capacity to suppress effector T‐cell responsiveness, because an increased nickel reactivity could still be detected in these cocultures. None of the isolated CD4⁺CD25⁺ cells, either isolated from healthy controls or allergic patients, produced IL‐10 in response to nickel. Overall, these results support the view that CD4⁺CD25⁺ cells can control the activation of nickel‐specific effector T cells in non‐allergic individuals, whereas this regulatory capacity is impaired in allergic patients. To investigate the presence of allergen‐specific regulatory T cells in truly naïve, non‐sensitized individuals, T‐cell reactivity should also be studied with non‐environmental contact allergens, such as para‐phenylenediamine.     

抑郁焦虑情绪对复发性尖锐湿疣患者外周血调节性T细胞水平的影响

目的: 确定抑郁焦虑情绪对CA复发和细胞免疫的影响。方法: 采用抑郁自评量表（SDS）和焦虑自评量表(SAS)对100例初诊CA进行问卷调查,评估其情绪状态。SDS>50和/或SAS>50为抑郁焦虑者,其余患者为非抑郁焦虑者。选择健康体检32人为对照。流式细胞技术测定外周血Treg细胞水平。随后观察两组CA治疗后的复发率。结果: CA组外周血Treg细胞水平为(8.03±2.14)%,显著高于正常对照组（5.28±1.21）% （P<0.05）;抑郁焦虑组外周血Treg细胞水平为（8.64±2.12）%,显著高于无抑郁焦虑组（7.55±2.05）% （P<0.05）。治疗后抑郁焦虑组第1、2、3、6个月的复发率均高于无抑郁焦虑组。结论:抑郁和/或焦虑情绪可加剧CA患者细胞免疫功能的抑制,导致CA更易复发。     

外周血及蜕膜组织中CD4~+ CD25~+调节性T细胞比例和炎症因子原因不明复发性流产的关系研究

目的:探究外周血及蜕膜组织中CD4~+CD25~+调节性T细胞比例以及相关炎症因子对原因不明复发性流产的影响。方法:选取2014年5月至2015年5月就诊于我院门诊的原因不明复发性流产(URSA)患者90例为观察组,并选取同期在妇产科门诊手术室的行人工流产手术的正常患者90例为对照组;分析两组患者的一般资料、外周血及蜕膜组织中CD4~+CD25~+调节性T细胞比例、调节性T细胞特异调节因子Foxp3和EBi3的表达以及血清炎症因子(包括TNF-α、IL-2、IL-6、IL-10)水平,研究其相关性。结果:与正常妊娠组相比,反复流产组外周血及蜕膜组织中Treg细胞比例明显降低,Foxp3和EBi3因子mRNA的表达水平明显降低,且无论URSA患者或者是正常对照组,蜕膜中各因子的表达均高于外周血中的表达,外周血TNF-α、IL-2水平明显升高,而IL-6和IL-10水平要明显降低,差异有统计学意义(P0.05)。结论:蜕膜中各因子的表达均高于外周血中的表达提示妊娠过程的主要免疫反应部位是"母-胎界面",Treg细胞比例、Treg细胞因子和Th2型因子IL-6、IL-10水平可能参与妊娠的维持,调控"母-胎界面"局部免疫耐受。对于复发性流产的发病过程具有一定的影响作用。     

卵巢癌患者腹水及外周血CD4~+CD25~+调节性T细胞含量及抑制功能的研究

目的:分析并探讨卵巢癌患者腹水及外周血CD4~+CD25~+调节性T细胞含量及抑制功能。方法:选取2012年2月至2015年2月期间在我院和宁波市鄞州第二医院接受治疗的卵巢癌患者56例,采集腹水标本56例,外周血标本56例。使用流式细胞术检测CD4~+CD25~+调节性T细胞(Treg)的表达。采用1.5μmol/L的羧基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)标记CD4~+CD25-T细胞,观察CFSE标记效果,计算抑制率。结果:研究结果显示,56例腹水患者CD4~+CD25~+T cells/CD4~+T cell为(28.34±13.27)%,56例外周血标本CD4~+CD25~+T cells/CD4~+T cell为(14.56±4.36)%。腹水和外周血标本CD4~+CD25~+T cells/CD4~+T cell含量有显著差异(P0.05)。腹水Treg抑制功能比外周血Treg强,经统计学检验,差异具有统计学意义(P0.05)。结论:腹水Treg含量及抑制功能比外周血Treg强,提示卵巢癌腹腔内相对容易发生免疫逃逸,而Treg升高可能参与促进肿瘤复发。     

The Clinical Features and Pathophysiology of Acute Radiation Dermatitis in Patients Receiving Tomotherapy

Background

Radiation therapy (RT) including tomotherapy has been widely used to treat primary tumors, as well as to alleviate the symptoms of metastatic cancers.

Objective

The primary purpose of this study was to examine the characteristics of the clinical features and pathophysiological mechanisms associated with acute radiation dermatitis in cancer patients that received tomotherapy, and compare the results to patients treated by conventional radiation therapy.

Methods

The study population consisted of 11 patients that were referred to the dermatology department because of radiation dermatitis after receiving tomotherapy; all patients were evaluated for clinical severity. The patients were assessed and identified using the National Cancer Institute Common Toxicity Criteria version (CTC) 3.0. We performed biopsies of the skin lesions that were examined for apoptosis using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labelling (TUNEL) assay and stained immunohistochemically with monoclonal antibodies to CD8, CD4 and TGF-β. As a positive control, patients with radiation dermatitis treated with conventional radiation therapy were also studied.

Results

The results of the clinical features of the skin of tomotherapy patients were the following: grade 1 (36%), grade 2 (55%) and other changes (9%). Among the population that had skin lesions due to acute radiation dermatitis, the mean number of positive cells per high power field (HPF) was the following: there were 30.50±7.50 TUNEL-positive cells, 34.60±12.50 CD8+ T cells, 5.19±3.17 CD4+ T cells and 9.95±1.33 TGF-β positive cells measured per HPF. The mean number of positive cells per HPF for the patients that received conventional radiation therapy was: TUNLEL-positive cells in 7.5±1.64, CD8-, CD4- and TGF-β-positive cells in 12.50±3.73, 3.16±1.47, 6.50±1.97.

Conclusion

We found that the number of TUNEL-positive cells and CD8+ T cells were higher in the lesions of patients receiving tomotherapy compared to the lesions of the patients receiving conventional radiation therapy. These findings suggest that tomotherapy without dose modification may cause significantly more severe forms of radiation dermatitis by apoptosis and cytotoxic immune responses than conventional radiation therapy.     

Involvement of granulysin-producing T cells in the development of superficial microbial folliculitis

BACKGROUND: Granulysin is a recently identified antimicrobial protein expressed on cytotoxic T cells, natural killer (NK) cells and NKT cells. It has been shown that granulysin contributes to the defence mechanisms against mycobacterial infection. Superficial microbial folliculitis is a common skin disease. In a previous report, we showed that, as a first line of defence, alpha-defensin, a human neutrophil peptide, and beta-defensin (human beta-defensin-2) were expressed in infiltrating neutrophils and in lesional epidermal keratinocytes, respectively, in superficial folliculitis. As we also observed many infiltrating lymphocytes in lesional dermis, we hypothesized that infiltrating lymphocytes may possess antimicrobial substances, such as granulysin, and play a role in the defence mechanism as a second line of defence. OBJECTIVES: Seven specimens of superficial microbial folliculitis diagnosed clinically and histologically were examined by means of immunohistochemistry. To identify the phenotype of cells expressing granulysin, confocal laser microscopic examination was performed. RESULTS: A dense lymphoid cell infiltrate was observed in pustules, in the perivascular regions. A large number of these lymphoid cells were positive for granulysin. Phenotypically, cells consisted of CD3+ T cells, CD8+ T cells and UCHL-1+ T cells. CD20+ cells and CD56+ cells were not observed. Microscopic examination with a confocal laser showed that the lymphocytes producing granulysin were CD3+ and CD4+ T cells but not CD8+ T cells. CONCLUSIONS: We showed that many granulysin-bearing T cells infiltrated affected follicles and perilesional dermis in superficial microbial folliculitis. However, few granulysin-positive lymphoid cells were observed in sterile pustular lesions. Our observations indicated that adaptive immunity such as granulysin, a lymphocyte-produced antimicrobial protein, may play an important role in the cutaneous defence mechanism.