单双标记技术对T细胞亚群测定的影响

目的 探讨单、双标记技术对Ｔ细胞亚群测定的影响。方法 采用荧光抗体双标记、流式细胞仪对正常４人外周血细胞进行测定。结果 ＣＤ３＾＋ＣＤ４＾＋与ＣＤ４＾＋细胞比较，差异无显著性意义，ＣＤ３＾＋ＣＤ８＾＋与ＣＤ８＾＋比较，有显著性差异（Ｐ〈０．０５），ＣＤ３＾＋ＣＤ４＾＋／ＣＤ３＾＋ＣＤ８＾＋与ＣＤ８＾＋比较，差异有显著性意义（Ｐ〈０．０５）。结论 检测Ｔ细胞亚群应采用双标记技术。     

异位性皮炎的记忆性T细胞浸润及ICAM—1表达

为了解粘连分子在异位性皮炎炎症及免疫反应过程中的作用，对ＡＤ皮损部位细胞间粘连分子－１的表达作了研究。结果虽然正常皮肤表皮不表达ＩＣＡＭ－１但ＡＤ皮损处角朊细胞则局灶性表达ＩＣＡＭ－１，尤其在有严重单个核细胞浸润及表皮内淋细胞移入的部位。免疫表型研究表明，ＡＤ真皮浸润中ＣＤ４＾４＋／ＣＤｗ２９＾＋／ＣＤ４５ＲＡ＾－１记忆性Ｔ细胞占主导，推测它们可能通过分泌某些细胞因子而诱导角朊细胞表达ＩＣＡＭ－１     

细胞毒T细胞识别皮肤T细胞淋巴瘤的T细胞受体个体基型肽抗原

目的 确定细胞毒Ｔ细胞是否能识别由第三互补决定区特异基因编码的肽。方法 选择两例皮肤Ｔ细胞淋巴瘤（ＣＴＣＬ）患者（ＳＳ和ＡＲ）和ＣＤ４＾＋、Ｖβ８＾＋恶性Ｔ细胞作为测定肽序列的细胞，ＣＤ８＾＋非恶性Ｔ细胞作为杀伤或反应性细胞，和转化的淋巴母细胞样细胞作为主要组织相容性复合体（ＭＨＣ）抗原提呈的细胞。结果 患者ＳＳ的恶性Ｔ细胞受体（ＴＣＲ）β链的个体基因型肽可刺激ＣＤ８＾＋Ｔ细胞产生肿瘤坏死因子（Ｔ     

荨麻疹外周血T淋巴细胞及亚群的检测

对６０例荨麻疹患者用流式细胞仪检测外周血Ｔ淋巴细胞及其亚群并与２０例健康志愿者作对照。结果表明，荨麻疹患者ＣＤ＾＋３及ＣＤ＾＋４细胞明显降低，与健康对照组比较差异有非常显著性意义，其中以急性荨麻疹患者ＣＤ＾＋３、ＣＤ＾＋４改变最为明显。     

系统性红斑狼疮患者外周血淋巴细胞bcl-2的表达及临床意义

目的 为了探讨ｂｃｌ－２基因在系统红斑狼疮（ＳＬＥ）发病机制中的作用及临床意义。方法 应用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）检测３１例ＳＬＥ患者外周血单一核细胞（ＰＢＭＣ）ｂｃｌ－２ｍＲＮＡ表达水平和流式细胞仪双标记法分析其Ｔ、Ｂ细胞ｂｃｌ－２蛋白表达。结果 活动期ＳＬＥ患者ＰＢＭＣｂｃｌ－２ｍＲＮＡ表达水平明显升高,占５５．６％,且活动期ＳＬＥ患者ＣＤ３＾＋、ＣＤ４＾＋和ＣＤ８＾＋Ｔ细胞亚群Ｐ     

红斑狼疮皮损中浸润细胞免疫表型和角质形成细胞HLA-DR抗原表达的研究

应用抗ＨＬＡＤＲ、ＣＤ３、ＣＤ４、ＣＤ８、ＣＤ２０的单克隆抗体和ｓｔｒｅｐｔｒａｖｉｄｉｎｐｅｒｏｘｉｄａｓｅｓｔａｉｎｉｎｇ（ＳＰ）技术对１０名正常人皮肤,１６例ＳＬＥ皮损和１９例ＤＬＥ皮损进行了免疫组化研究。观察到正常人皮肤角质形成细胞未见ＨＬＡＤＲ抗原表达,而ＳＬＥ（６／１６）,ＤＬＥ（８／１９）皮损处角质形成细胞可以表达ＨＬＡＤＲ抗原。在ＳＬＥ、ＤＬＥ真皮内浸润细胞主要为Ｔ淋巴细胞（ＣＤ３＋浸润细胞）,且以ＴＨ细胞（ＣＤ４＋浸润细胞）占优势。另外,还发现在两种ＬＥ表皮角质形成细胞表达ＨＬＡＤＲ抗原处,真皮内可见ＣＤ３＋浸润细胞和激活的Ｔ淋巴细胞（ＨＬＡＤＲ＋浸润细胞）。讨论了ＬＥ皮损角质形成细胞ＨＬＡＤＲ抗原表达及其与病损内浸润细胞免疫表型的关系。ＬＥ皮损处ＨＬＡＤＲ＋角质形成细胞可能具有抗原递呈作用,而角质形成细胞异常表达ＨＬＡＤＲ抗原则可能与真皮内浸润单个核细胞或淋巴细胞释放的ＩＦＮα,ＴＮＦγ等有关。     

白癜风患者外周血T淋巴细胞亚群和sIL-2R水平变化的研究

采用ＡＰＡＡＰ法和双缺本夹心ＥＬＩＳＡ技术检测１５０例白癜风患者外周血Ｔ淋巴细胞亚群及可溶性白介素－２受体（ｓＩＬ－２Ｒ）水平。结果显示：（１）寻常型白癜风外周血ＣＤ３＾＋、ＣＤ４＾＋细胞数、ＣＤ４＾＋／ＣＤ８＾＋比值明显低于正常对照（Ｐ〈０．０１）,ｓＩＬ－２Ｒ活性显著性高于节段型白癜风和正常人（Ｐ〈０．０１）;节段型白癜风ＣＤ４＾＋、ＣＤ４＾＋／ＣＤ８＾＋与正常对照组比较也显著减少（Ｐ〈０．０     

原发性皮肤CD^＋30间变性大T细胞淋巴瘤1全

报告１例罕见的原发皮肤Ｔ细胞淋巴瘤。患者女，１８岁，头顶部不明原因的反复出现 结节状肿物。病理检查，免疫组织化学染色瘤细胞ＣＤ＾＋３０、ＣＤ＾＋３、ＬＣＡ＾＋、ＥＭＡ＾＋、ＣＥＡ＾－、ＣＤ２＾－２０。对肿瘤局部放射治疗效果满意。     

异位性皮炎等皮损内郎格罕细胞携带IgE分子的研究

用ＡＢＣ免疫过氧化酶技术和免疫荧光双标记技术，在异位性皮炎等皮肤冰冻切片上观察郎格罕细胞的分布及ＩｇＥ＾＋／ＯＫＴ６＾ＬＣ的存在；同时应用ＥＬＩＳＡ法测定了部分患者血清ＩｇＥ＾／ＬＫＴ６＾＋ＬＣ的存在。在血清ＩｇＥ水平不高的湿疹，变应性接触性皮炎等非ＡＤ皮损内也见到ＩｇＥ＾＋／ＯＫＴ６＾＋ＬＣ的存在。我们认为ＩｇＥ＾＋ＬＣ的存在并非对ＡＤ是特异的。     

环孢素A及雷公藤内酯醇对CD4^＋T细胞,角质形成细胞和Hela细 …

目的 比较环孢素Ａ（ＣｙＡ）和雷公藤提取物Ｔ０对ＣＤ４＾＋Ｔ细胞，角质形成细胞和Ｈｅｌａ细胞增殖的影响。方法 采用人外周血单一核细胞（ＰＢＭＣ），人表皮角质形成细胞和Ｈｅｌａ细胞培养，＾Ｈ－ＴｄＲ掺入法测定药物对细胞ＤＮＡ合成的影响，结果Ｔ０和ＣｙＡ都能抑制ＣｏｎＡ诱导的人外周血Ｔ淋巴细胞增殖。Ｔ０在体外可抑制生长于低无血清角质形成细胞培养基中的正常人角持形成细胞和Ｈｅｌａ细胞株的生长，而ＣｙＡ     

Immunocompetent cells of fixed drug eruption

The positive provocation test reactions of the skin of six patients with fixed drug eruption (FDE) were studied from timed skin biopsies taken between 2 hours and 9 days after the appearance of FDE. Monoclonal antibodies to the following immunocompetent cell surface epitopes were used: T3, T4, T6, T8, T9, M1, Ia1, Drc, Leu7 and B cell. The dermal infiltrate comprised 60-80% of T lymphocytes at all the times studied. Cells with T4 and T8 epitopes were displayed in similar numbers. A transient decrease in the number of T6+ cells of the epidermis could be detected with a simultaneous and also transient increase of the T6+ cells in the dermis, which suggests a possible traffic of Langerhans' cells from the epidermis to the dermis. The epidermal Ia1+ cells showed changes similar to but less marked than the T6+ cells. The number of the dermal Ia1+ cells increased continuously. In the late biopsies these Ia1+ cells comprised up to 90% of the infiltrating cells. Except for the finding of a reduction of T6+ and Ia1+ epidermal cells, the cellular kinetics of FDE are similar to those seen in both cutaneous immunological and irritant reactions.     

Successful desensitization to fixed drug eruption: the presence of CD25+CD4+ T cells in the epidermis of fixed drug eruption lesions may be involved in the induction of desensitization

BACKGROUND: Fixed drug eruption (FDE) is a distinct type of drug-induced eruption, in which intraepidermal CD8+ T cells in the lesional skin are the final effector cells in the epidermal injury of FDE. Desensitization is a unique approach for the management of drug eruption, which has been reported to be effective in treating FDE. However, the mechanisms underlying desensitization to FDE are quite unknown. OBJECTIVE AND METHODS: We reported a case of successful desensitization to allopurinol-induced FDE. To clarify the mechanisms underlying desensitization to FDE, we examined the phenotype of T cells in the epidermis of FDE lesions before and after desensitization using flow cytometry. RESULTS: The overwhelming majority of intraepidermal T cells in the FDE lesion before desensitization consisted of CD8+ T cells, whereas a significant number of CD25+CD4+ T cells were present in the epidermis of FDE lesions after desensitization. CONCLUSION: The presence of CD25+CD4+ T cells in the epidermis of FDE lesions may be involved in the induction of desensitization to FDE.     

二期梅毒患者外周血Th1/Th2及Tc1/Tc2的研究

目的探讨CD4+T细胞(Th细胞)和CD8+T细胞(Tc细胞)亚群在梅毒发病机理中的作用及其相互关系。方法采用流式细胞术检测经三色荧光抗体染色的二期梅毒患者外周血CD4+/IFN-γ+(Th1),CD4+1/IL-4+(Th2),CD4+/IFN-γ+/IL-4+(Tho),CD8+/IFN-γ+(Tc1),CD8+/IL-4+(Tc2),CD8+/IFN-γ+/IL-4+(Tc0)细胞含量。结果梅毒组Th1细胞含量、Th1/Th2,Tc1/Tc2比值均明显低于正常对照组(P<0.01),而Tc0和Tc2细胞含量显著高于正常对照组,其他细胞两组间无显著性差异(P>0.05)。结论二期梅毒患者Th1/Th2及Tc1/Tc2比值失衡,可能是机体不能完全清除梅毒螺旋体造成其潜伏体内形成长期感染的重要因素之一。     

复发性生殖器疱疹患者外周血IL-12与Th1/Th2细胞因子的检测

目的检测复发性生殖器疱疹(RGH)患者不同病期外周血CD4+T细胞内IL-12,IFN-,γIL-4的水平,探讨IL-12,Th1与Th2亚群在疾病中的可能作用。方法应用流式细胞仪对20例发作期、15例恢复期RGH患者和15名健康人外周血CD4+T细胞IL-12,IFN-γ和IL-4进行检测。结果发作期患者外周血IFN-γ+-CD4+T细胞百分率显著低于正常对照组(P<0.05),IL-4+-CD4+T细胞百分率明显高于正常对照组(P<0.01),Th1/Th2比值显著低于正常对照组(P<0.01),同时IL-12+-CD4+T细胞百分率显著降低(P<0.01)。恢复期患者外周血IL-12+-CD4+T细胞百分率仍显著低于正常(P<0.05)。结论RGH患者存在Th1/Th2比例失衡和IL-12水平低下,而后者可能是导致Th1/Th2比例失衡和病情反复发作的重要原因。     

T-cell subsets in drug-induced toxic epidermal necrolysis. Possible pathogenic mechanism induced by CD8-positive T cells

A patient with bromisovalum-induced toxic epidermal necrolysis showed pronounced delayed hypersensitivity to bromisovalum by patch testing. Biopsy specimens from the cutaneous lesion and the site of the positive patch test reaction were analyzed and compared immunohistologically. The findings were similar: most of the mononuclear cells disposed along the dermoepidermal junction and migrating into the epidermis were CD8-positive lymphocytes, whereas the dermal inflammatory infiltrates were composed predominantly of CD4-positive lymphocytes. This case showed the potential usefulness of patch testing in evaluating cases of toxic epidermal necrolysis. We believe that delayed hypersensitivity plays a crucial role in the development of drug-induced toxic epidermal necrolysis. Furthermore, potential effector cells with phenotypic characteristics of CD8-positive lymphocytes (suppressor/cytotoxic T cells) seem to represent important mediators of the epidermal damage of the cutaneous lesion in our case.     

Expansion of IL-10-producing CD4+ and CD8+ T cells in fixed drug eruption

BACKGROUND: A severe form of fixed drug eruption (FDE) clinically and histologically mimics toxic epidermal necrolysis (TEN) but, unlike TEN, resolves spontaneously upon withdrawal of the causative drug. OBJECTIVE AND METHODS: We reported a case of a severe FDE caused by mefenamic acid that spontaneously resolved without use of systemic corticosteroids. To clarify the phenotype of the T cells responsible for clinical resolution of FDE, we kinetically examined gamma-interferon (IFN-gamma), interleukin (IL)-2, IL-4 and IL-10 production by peripheral blood T cells of the patient before and after oral challenge with the causative drug using flow cytometry. RESULTS: We found that the proportions of CD4+ and CD8+ T cells capable of producing IFN-gamma and IL-4 remained unchanged after challenge, while those of CD4+ and CD8+ T cells capable of producing IL-10 dramatically increased after challenge. The frequency of CD8+ T cells capable of producing IL-2 decreased after challenge. CONCLUSION: These results suggest that expansion of IL-10-producing CD4+ and CD8+ T cells may be responsible for spontaneous resolution of a severe form of FDE.     

Expression of cutaneous lymphocyte-associated antigen and TIA-1 by lymphocytes in pityriasis lichenoides et varioliformis acuta and lymphomatoid papulosis: immunohistochemical study

BACKGROUND: Pityriasis lichenoides et varioliformis acuta (PLEVA) and lymphomatoid papulosis (LyP) are benign self-healing cutaneous eruptions that may be clinically and histologically similar. The purposes of this study were to evaluate immunohistological characteristics of PLEVA and LyP and to investigate whether Epstein-Barr virus (EBV) may be present in PLEVA and LyP. METHODS: We performed an immunohistochemical staining in 12 cases of PLEVA and 8 cases of LyP using nine antibodies for CD3, CD4, CD8, CD30, CD45RO, CD56, CD79, cutaneous lymphocyte-associated antigen (CLA), and TIA-1. In situ hybridization was performed using fluorescein-conjugated oligonucleotide probes for EBV early regions (EBER). RESULTS: In PLEVA, immunohistochemical studies revealed that infiltrated lymphocytes consisted of mainly CD3-positive (5+), CD8-positive (4+ to 5+), CLA-positive (4+ to 5+) T cells and partly CD79 positive (+ to 2+) B cells. CD4-positive T cells were less than 25%. In LyP, immunohistochemical studies revealed that infiltrated lymphocytes consisted of partly CD3-positive (5+), CD8-positive (2+ to 3+), CLA-positive (3+ to 4+) T cells and partly CD79-positive (2+ to 3+) B cells. CD4-positive T cells were less than 10%. CD8 and CLA were more strongly expressed in PLEVA than in LyP. CD30 was strongly expressed in LyP but not expressed in PLEVA. CD79 was more expressed in LyP than in PLEVA. TIA-1 was not expressed in any cases. In situ hybridization using antisense EBER probe showed negative reaction in all cases. CONCLUSIONS: Immunohistochemical stains for CD8, CD30, CD79 and CLA may be valuable tools in the differential diagnosis between PLEVA and LyP. TIA-1 was negative in LyP, which means cytotoxic cells may not be implicated in the pathogenesis of LyP. It was a contradictory result to the previous results. The absence of EBV in PLEVA and LyP suggests that this virus may not be operative in the pathogenesis of these diseases. These results suggest that LyP and PLEVA are separate disorders, thus accounting for their variable prognosis.     

Increased number of OKT6-positive dendritic cells in the hair follicles of patients with alopecia areata

In 6 patients with untreated alopecia areata in the progressive stage, 6 in the stationary stage, and 6 normal individuals as controls, an in situ analysis of OKT6-positive dendritic cells in hair follicles, and peribulbar and intrabulbar infiltrates was performed using the avidin-biotin-peroxidase method with monoclonal antibodies. In controls, OKT6-positive dendritic cells were distributed only in the upper portions of hair follicles and were not observed in the bulbar area, and the percentage of these cells among all epithelial cells of the hair follicles was 1.0 +/- 0.1% (mean +/- SE). In stationary-stage patients, the distribution and the percentage of positive cells were the same as those for the controls (1.1 +/- 0.1%). In the progressive stage, however, positive cells were distributed in both the upper portions of the hair follicles and the bulbar area, and the percentage of positive cells (4.9 +/- 0.3%) was significantly higher than that of controls. Staining for T, B lymphocytes and T cell subsets in the peribulbar infiltrates revealed a predominance of OKT4-positive cells (the OKT4/OKT8 ratio was from 3:1 to 4:1). This indicates that the number of OKT6-positive dendritic cells increases in the hair follicles of progressive alopecia areata and that these cells may play an important role in cooperation with T cells in the pathogenesis of alopecia areata.     

Tumor-infiltrating T cells in primary cutaneous B-cell lympho-proliferative disorders

BACKGROUND: Tumor-infiltrating lymphocytes (TILs) are considered to play an important role in the antitumoral immune response. The presence and percentage of CD8-positive tumor-infiltrating T cells have been shown to correlate with differentiation and prognosis in various neoplasms. The aim of this study was to determine the number of CD8-positive T cells in various primary cutaneous B-cell lymphoproliferative disorders and to evaluate its correlation with the histological type of tumor. METHODS: Fifty-three lesions were examined by immunohistochemistry with antibodies targeting CD3, CD4, CD8 and TIA-1. Thirty-two lesions had been diagnosed as primary cutaneous B-cell lymphomas (CBCL) and 21 as B-cell pseudolymphomas (B-PSL). CBCLs included 15 follicular lymphomas (FL), 6 marginal zone lymphomas (MZL), and 11 diffuse large B-cell lymphomas (LCL). The number of CD8-positive cytotoxic T cells was determined by computer-assisted morphometrical microscopy. RESULTS: No significant difference could be detected in the density of CD8-positive T cells in B-PSL (101/105 microm(2)), FL (110/105 microm(2)), and MZL (122/105 microm(2)). In contrast, the number of CD8-positive cells (55/105 microm(2)) in LCL was significantly lower (p<0.01) compared to B-PSL, FL and MZL. CONCLUSIONS: In summary the number of CD8-positive T cells in B-cell lymphoproliferative disorders differs in regard to tumor type and differentiation with lowest numbers in diffuse large B-cell lymphomas. However, due to an overlap of the number of TILs, this parameter cannot be employed as a diagnostic parameter for individual cases.     

云南省2152例HIV/AIDS流行特征及CD_4~+CD_8~+检测结果分析

目的了解云南省艾滋病流行特征。方法分析流行病学资料,应用流式细胞计数仪和三标TruCount荧光标记单克隆抗体试剂盒对2 152例H IV感染者/AIDS患者进行CD4、CD8检测。结果10~19岁组H IV/AIDS中女性占61.5%,暗娼占28.2%,远远大于其他年龄组。通过性途径传播的在50岁以上组中所占比例最大(52.6%),10~19岁组次之,占41.0%;2 144例中CD4+<200个/mL的有356例(16.6%),其中50岁以上龄组<200个/m l的占36.8%,10~19岁组中>350个/m l的占82.1%;AIDS患者CD4+、CD4+/CD8+低于H IV感染者(P<0.001和P<0.001),2 144例H IV/AIDS中CD4+/CD8+≥1的有41例,其中女性占51.2%,异性传播感染的占31.7%。结论目前云南省艾滋病病毒感染者虽然仍以注射吸毒人群为主,但性传播和母婴传播危险性加大。CD4+、CD4+/CD8+与临床进程明显相关,随着疾病的进展,CD4+、CD4+/CD8+持续降低。发现部分感染者CD4+/CD8+≥1,未倒置情况是否提示该组感染时间较短,其流行病学分析资料是否可为艾滋病的控制提供依据还有待进一步研究。