PHA对人扁桃体T淋巴细胞亚群变化及活化抗原表达的效应

应用FITC和PE双色荧光试剂并经流式细胞计分析了人扁桃腺T淋巴细胞亚群组成,以及用PHA刺激后T淋巴细胞亚群的变化和活化抗原的表达情况。结果表明:1)人扁桃体T淋巴细胞以CD4~+细胞占优势,CD4~+与CD8~+细胞比值约为5.32±0.55;2),经PHA刺激培养72小时,CD4~+CD8~+细胞明显增加;3)经PHA刺激培养后,IL-2受体(CD25)和HLA-DR抗原表达增加。本实验结果为深入研究人扁桃体T淋巴细胞的表型变化提供了客观指标,对进一步探讨其在免疫系统中的作用具有一定的意义。     

人类T淋巴细胞亚群

根据T淋巴细胞的功能,以前曾将其分为Ts,Th、Tc、Ta及Td等5个亚群。近来根据表面Ig、Fc受体的不同,又把人类T淋巴细胞分为两个亚群:具有IgM Fc受体者称Tμ细胞,具有IgG Fc受体者称Tγ细胞Moretta等报告,人类外周血中多数T淋巴细胞具有IgM Fc受体,少数     

人类T淋巴细胞亚群

在人类免疫生物学,特别是T细胞亚群的研究中,细胞表面标志物的表露及这些标志物在细胞亚群功能性状鉴定中发挥的作用已成为极为热门的研究领域。一般认为T细胞具有绵羊红细胞受体(E受体)。这一表面标志物可用于总T细胞群的鉴定。体外T细胞功能测定中已经证明具有T细胞功能的细胞主要是形成E玫瑰花的细胞群,因此这一标志物是很有实用价值的。但是不能认为T细胞就是具有E受体的细胞。虽然我们知道所有具有T细胞功能的细胞都位于E玫瑰花细胞群中,但这并不意味着每一个E玫瑰花细胞必定是T细胞。目前已知某些T细胞结合E的亲合力大大低于     

T淋巴细胞亚群及其临床意义

许多研究工作表明,T淋巴细胞不仅是细胞免疫的效应细胞,而且在免疫反应的调节上具有重要的作用。T细胞的免疫调节作用是由T细胞的亚群,尤其是功能上不同的亚群来完成的。因而,T细胞亚群的研究和检测在许多疾病的免疫发病学研究上,以及在免疫性疾病和免疫病理过程的防治研究上都有重要的意义。     

脐血T淋巴细胞亚群的研究

本文用３Ｈ—ＴｄＲ和ＡＰＡＡＰ法研究了脐血淋巴细胞转化功能（ＬＴ）、脐血血清的特性、混合淋巴细胞反应（ＭＬＣ）及Ｔ细胞亚群，结果表明脐血淋巴细胞对丝裂原的反应与正常成人外周血淋巴细胞无明显差异；脐血血清对ＰＨＡ和ＣｏｎＡ诱导的淋巴细胞转化有抑制作用；脐血之间、脐血与成人外周血之间的ＭＬＣ反应均较成人外周血之间的ＭＬＣ反应弱，且有显著性差异（Ｐ＜０．０５）。脐血细胞表型尚未成熟，脐血ＣＤ３＋细胞数减低，ＣＤ４＋和ＣＤ８＋细胞数及其比率近似于成人，细胞数之和大于ＣＤ３＋细胞数。     

人淋巴细胞亚群的异种抗血清

曾有过许多关于人的T、B细胞特异的异种抗血清的报告.T细胞抗血清主要是用胸腺细胞制备,再用B细胞(如慢性淋巴细胞性白血病细胞)来吸收,使之更具有特异     

结肠癌浸润T淋巴细胞亚群的临床意义

目的目前有报道认为淋巴细胞浸润与结肠癌(CRC)病理学变化相关,但CRC病人体内的淋巴细胞亚群各自的临床意义不清楚,本研究就此进行了探讨。方法我们使用了组织芯片和免疫组化技术检测67例CRC病人肿瘤组织中心区和边缘区CD4+和CD8+细胞密度,并分析其临床意义。结果我们发现,肿瘤浸润淋巴细胞亚群与病人的年龄、肿瘤大小、肿瘤的分期/分级不相关,且CD4+T细胞在不同分期/分级病人的肿瘤中心区和边缘区均无显著性差异。但是,与M1级病人相比,M0级病人肿瘤边缘区有更高的CD8+T细胞密度,而肿瘤中心区CD8+T细胞在两者间无差别。结论肿瘤边缘区高密度的CD8+T细胞与低肿瘤转移率相关。结果还提示对肿瘤浸润淋巴细胞亚群的分析有利于深入理解免疫系统对肿瘤的作用及其机制,且对开发相关干预措施提供了线索。     

白血病时T淋巴细胞亚群的改变

白血病时Ｔ淋巴细胞亚群的改变毛玉文，尤育初，钱林法，李小雯，张红华，焦红近年来的研究表明，淋巴细胞不仅是免疫活性细胞，而且也参与机体的造血过程，且多数血液病的发展均伴有淋巴细胞亚群分布的改变［１～５］。因此，检测和分析淋巴细胞亚群的分布，对于评估血液...     

癫痫患者T淋巴细胞亚群的检测

<正> 癫痫患者存在免疫功能异常。本文检测癫痫患者的外、周血T淋巴细胞亚群以进一步探讨其免疫状态。 材料和方法 60例癫痫患者由我院神经科,经临床和脑电图确诊。男34例,女26例;年龄2～62岁,平均19.2岁;病程4周～40年;全身性发作型43例,部分性发作型17例;未服抗癫痫药物24例,服用抗癫痫药物36例。对照组30例系本院职工及临床确诊的神经衰弱患者。采用武汉生物制品研究所提供的抗     

免疫酶法检测人外周血T淋巴细胞亚群的初步探讨

用抗人淋巴细胞单克隆抗体检测外周血淋巴细胞亚群的主要方法很多。本文采用免疫酶法检测了人外周血T淋巴细胞亚群,并与间接免疫荧光法进行了对比,现将结果报道如下。 材 料 和 方 法 一、免疫酶法检测人外周血T淋巴细胞亚群(0KT_3ODT_OKT_3)试剂盒由中国科学院武汉病毒研究所供给。荧光法单克隆抗体(OKT_3OKT_4OKT_3)由卫生部武汉生物制品研究所提供。 二、外周血淋巴细胞的分离:取正常人外周血3ml,加肝素抗凝。按常规方法分     

Evaluation of T cell immune responses in multi-drug-resistant tuberculosis (MDR-TB) patients to Mycobacterium tuberculosis total lipid antigens

Mycobacterium tuberculosis lipid antigens produce significant T cell responses in healthy tuberculin reactor [purified protein derivative (PPD-positive] individuals. In the present study, proliferation and interferon (IFN)-gamma/interleukin (IL)-4 responses were analysed to M. tuberculosis total lipid antigens in T lymphocytes from 25 patients with multi-drug-resistant tuberculosis (MDR-TB). The obtained results were compared with those of 30 asymptomatic healthy PPD-positive and 30 healthy tuberculin skin test negative (PPD-negative) subjects. Peripheral blood mononuclear cells (PBMCs) and T cells (CD4(+) and CD8(+)) were stimulated using autologous immature dendritic cells. Proliferation responses were assessed using 3-{4,5-dimethylthiazol-2-yl}-2,5 diphenyl tetrazolium bromide (MTT). IFN-gamma/IL-4 concentrations in the supernatant of the CD4(+) and CD8(+)T cells were measured by enzyme-linked immunosorbent assay. Proliferation assay showed that the peripheral blood mononuclear cells and CD4(+) T cells from the MDR-TB patients responded significantly less to the M. tuberculosis total lipid antigens than to the CD4(+) T cells in the PPD-positive subjects. Total lipid antigen-specific proliferative responses in the CD8(+) T cells from the MDR-TB patients were minimally detected and the responses were similar to those of the PPD-positive subjects. IFN-gamma production by the CD4(+) T cells stimulated by total lipid antigens from the MDR-TB patients was decreased significantly compared with the PPD-positive individuals, whereas IL-4 production in the patients was elevated. IFN-gamma and IL-4 production in the CD8(+) T cells of the MDR-TB patients was similar to those of the PPD-positive subjects. In conclusion, it is suggested that stimulated CD4(+) T cells by M. tuberculosis total lipid antigens may be shifted to T helper 2 responses in MDR-TB patients.     

Characterization of human cellular immune responses to novel Mycobacterium tuberculosis antigens encoded by genomic regions absent in Mycobacterium bovis BCG

Comparative genomics has identified several regions of differences (RDs) between the infectious Mycobacterium tuberculosis and the vaccine strains of Mycobacterium bovis BCG. We aimed to evaluate the cellular immune responses induced by antigens encoded by genes predicted in 11 RDs. Synthetic peptides covering the sequences of RD1, RD4 to RD7, RD9 to RD13, and RD15 were tested for antigen-induced proliferation and secretion of Th1 cytokine, gamma interferon (IFN-gamma), by peripheral blood mononuclear cells (PBMC) obtained from culture-proven pulmonary tuberculosis (TB) patients and M. bovis BCG-vaccinated healthy subjects. Among the peptide pools, RD1 induced the best responses in both donor groups and in both assays. In addition, testing of TB patients' PBMC for secretion of proinflammatory cytokines (tumor necrosis factor alpha [TNF-alpha], interleukin 6 [IL-6], IL-8, and IL-1beta), Th1 cytokines (IFN-gamma, IL-2, and TNF-beta), and Th2 cytokines (IL-4, IL-5, and IL-10) showed differential effects of RD peptides in the secretion of IFN-gamma and IL-10, with high IFN-gamma/IL-10 ratios (32 to 5.0) in response to RD1, RD5, RD7, RD9, and RD10 and low IFN-gamma/IL-10 ratios (<1.0) in response to RD12, RD13, and RD15. Peptide-mixing experiments with PBMC from healthy subjects showed that secretion of large quantities of IL-10 in response to RD12 and RD13 correlated with inhibition of Th1 responses induced by RD1 peptides. In conclusion, our results suggest that M. tuberculosis RDs can be divided into two major groups--one group that activates PBMC to preferentially secrete IFN-gamma and another group that activates preferential secretion of IL-10--and that these two groups of RDs may have roles in protection against and pathogenesis of TB, respectively.     

Human immune response to Mycobacterium tuberculosis antigens.

Little is known about the immunodominant or protective antigens of Mycobacterium tuberculosis in humans. Cell-mediated immunity is necessary for protection, and healthy tuberculin-positive individuals are relatively resistant to exogenous reinfection. We compared the targets of the cell-mediated immune response in healthy tuberculin-positive individuals to those of tuberculosis patients and tuberculin-negative persons. By using T-cell Western blotting (immunoblotting) of nitrocellulose-bound M. tuberculosis culture filtrate, peaks of T-cell blastogenic activity were identified in the healthy tuberculin reactors at 30, 37, 44, 57, 64, 71 and 88 kDa. Three of these fractions (30, 64, and 71 kDa) coincided with previously characterized proteins: antigen 6/alpha antigen, HSP60, and HSP70, respectively. The blastogenic responses to purified M. tuberculosis antigen 6/alpha antigen and BCG HSP60 were assessed. When cultured with purified antigen 6/alpha antigen, lymphocytes of healthy tuberculin reactors demonstrated greater [3H]thymidine incorporation than either healthy tuberculin-negative controls or tuberculous patients (8,113 +/- 1,939 delta cpm versus 645 +/- 425 delta cpm and 1,019 +/- 710 delta cpm, respectively; P less than 0.01). Healthy reactors also responded to HSP60, although to a lesser degree than antigen 6/alpha antigen (4,276 +/- 1,095 delta cpm; P less than 0.05). Partially purified HSP70 bound to nitrocellulose paper elicited a significant lymphocyte blastogenic response in two of six of the tuberculous patients but in none of the eight healthy tuberculin reactors. Lymphocytes of none of five tuberculin-negative controls responded to recombinant antigens at 14 or 19 kDa or to HSP70. Antibody reactivity generally was inversely correlated with blastogenic response: tuberculous sera had high titer antibody to M. tuberculosis culture filtrate in a range from 35 to 180 kDa. This is the first systematic evaluation of the human response to a panel of native and recombinant antigens in healthy tuberculin reactors and tuberculous patients. Antigens which stimulated prominent lymphocyte blastogenic responses were identified in seven fractions on T-cell Western blot analysis. Two of these may represent previously characterized proteins; the others may contain immunodominant proteins that will require further characterization.     

T-cell responses to CD1-presented lipid antigens in humans with Mycobacterium tuberculosis infection

CD1-restricted presentation of lipid or glycolipid antigens derived from Mycobacterium tuberculosis has been demonstrated by in vitro experiments using cultured T-cell lines. In the present work, the frequency of T-cell responses to natural mycobacterial lipids was analyzed in ex vivo studies of peripheral blood lymphocytes from human patients with pulmonary tuberculosis, from asymptomatic individuals with known contact with M. tuberculosis documented by conversion of their tuberculin skin tests, and from healthy tuberculin skin test-negative individuals or individuals vaccinated with Mycobacterium bovis BCG. Proliferation and gamma interferon enzyme-linked immunospot assays using peripheral blood lymphocytes and autologous CD1(+) immature dendritic cells revealed that T cells from asymptomatic M. tuberculosis-infected donors responded with significantly greater magnitude and frequency to mycobacterial lipid antigen preparations than lymphocytes from uninfected healthy donors. By use of these methods, lipid-antigen-specific proliferative responses were minimally detectable or absent in blood samples from patients with active tuberculosis prior to chemotherapy but became detectable in blood samples drawn 2 weeks after the start of treatment. Lipid antigen-reactive T cells were detected predominantly in the CD4-enriched T-cell fractions of circulating lymphocytes, and anti-CD1 antibody blocking experiments confirmed the CD1 restriction of these T-cell responses. Our results provide further support for the hypothesis that lipid antigens serve as targets of the recall response to M. tuberculosis, and they indicate that CD1-restricted T cells responding to these antigens comprise a significant portion of the circulating pool of M. tuberculosis-reactive T cells in healthy individuals with previous exposure to M. tuberculosis.     

结核病T细胞亚群的研究进展

T淋巴细胞亚群的数量和功能变化与结核病的病情进展和转归密切相关,探讨结核杆菌感染机体时T淋巴细胞亚群免疫应答及其影响因素,对结核病预防、诊治有重要意义。     

Kinetics of T lymphocyte responses to persistent antigens

Long term sequential study of immune responses in the same individuals is difficult from the time commitment required and the problem of maintaining enough subjects to provide for comparative analysis. We closely studied one hundred women with silicone mammary devices through cross sectional analysis up to 26 years post implantation and a similar sample of women to 6 years post explantation. The T cell index, calculated from tritiated thymidine incorporation during lymphoblast transformation, rose to a post implant peak at 10.5-12.0 years, falling progressively over the next 14.0-15.5 years to values indicative of probable immune quiescence. Post explantation, the index rose over the first 3 years and then sharply declined to within the range for unexposed controls. The shape of these time curves contains considerable information referent cell dynamics for both stimulatory and inhibitory factors and for demonstrating net group effects, appropriate to analysis in the cross sectional perspective. When a subset of four women was studied frequently and sequentially up to 8 years, an internal oscillatory pattern emerged, focusing attention on both the stimulatory and the inhibitory aspects of long term clonal expansion. IL-2 has stimulatory and inhibitory properties at different levels of production and is considered a prime candidate as the essential cytokine. The equations have details, however, which require exploration beyond any such provisional conclusion. The analytic process was aided by normalization of oscillatory data to eliminate subject variability and by Pareto optimization to assess the trend shown by normalization. Pareto analysis revealed two minimally coordinated oscillations, one over time and the other along net clonal expansion or decline of the siloxane specific T lymphocyte clone. The segments of the time related oscillation greatly exceeded the reaction times of cytokines currently known to be active in T cell regulation. Although the ultimate controlling factor(s) may be cytokine or chemokine combinations, the data are compatible with some more basic regulatory factor(s) of cell integrity, including limits on the number of cell divisions which can be sustained in long term immunopathic lesions, among other processes.     

In vitro cellular immune responses to complex and newly defined recombinant antigens of Mycobacterium tuberculosis

The immunological diagnosis and development of new antituberculosis vaccines require the characterization of Mycobacterium tuberculosis antigens inducing cell-mediated immune responses. In this study, we have tested peripheral blood mononuclear cells (PBMC) from tuberculosis (TB) patients (n = 43) and Bacille Calmette-Guérin (BCG)-vaccinated healthy subjects (n = 24) for in vitro cellular immune responses, as indicated by antigen-induced proliferation and interferon (IFN)-gamma secretion, in response to a panel of complex (culture filtrate and cell wall preparations) and single recombinant antigens (Mtb8.4, Mtb9.8, Mtb9.9, Mtb32A, Mtb39A, Mtb40, Mtb41 and Ag85B) of M. tuberculosis. The results of cellular responses showed that the majority (ranging from 70 to 98%) of TB patients and healthy donors responded to the complex antigens in antigen-induced proliferation and IFN-gamma secretion assays. However, when PBMC from the same groups of patients and healthy donors were tested with the recombinant antigens, TB patients showed strong recognition (>50% responders) of Mtb9.8 and Mtb39A in proliferation assays (median SI = 6.2 and 6.4, respectively) and of Mtb9.8, Mtb39A, Mtb40 and Ag85B in IFN-gamma assays (median delta IFN-gamma= 15.5, 10.8, 7.8 and 8.1 U/ml, respectively). BCG-vaccinated healthy donors showed weak (<30% responders) to moderate (31-50% responders) responses to all of the recombinant antigens in both assays. When PBMC of a subset of TB patients (n = 11) were tested for secretion of protective Th1 cytokines [IFN-gamma, tumour necrosis factor (TNF)-alpha and interleukin (IL)-12] and the immunosuppressive cytokine IL-10, the complex CF and CW antigens as well as the recombinant Mtb9.8, Mtb9.9, Mtb40 and Ag85B induced the secretion of both types of cytokines. On the other hand, Mtb41 induced only IL-10, while Mtb8.4, Mtb32Aand Mtb39A induced the secretion of one or more of Th1 cytokines, but not IL-10. In conclusion, the recombinant antigens inducing the secretion of Th1 cytokines could be useful as subunit vaccine candidates against TB.     

T lymphocyte subsets in human intestinal mucosa: the distribution and relationship to MHC-derived antigens

T lymphocytes in the normal human intestinal tract have been analysed in tissue sections by a double-marker immunofluorescence technique, combining antiserum to T lymphocyte antigen (HuTLA) with a monoclonal antibody detecting T cells of suppressor-cytotoxic phenotype (OKT8). The distribution of HLA-A -B, -C and Ia-like antigens in intestinal mucosa was also examined by a similar method. In small and large intestine 67 to 90% (mean 70%) of intraepithelial T lymphocytes were of suppressor-cytotoxic phenotype (OKT8+). In contrast, only 27 to 56% (mean 39%) of lamina propria T cells were OKT8+. Intestinal epithelial cells demonstrated strong membrane staining for HLA-A, -B, -C antigens. Ia-like antigens were detected on the epithelial cells of small intestinal villi, but not on colonic epithelial cells. Lamina propria macrophages expressed both HLA-A, -B, -C and Ia-like antigens, the latter having strong membrane and cytoplasmic fluorescence. The distribution of T cells with suppressor-cytotoxic or inducer phenotype in the intestinal epithelium and lamina propria may be related to the differential expression of Ia-like and HLA-A, -B, -C antigens in intestinal mucosa.     

T cell responses of normal individuals towards recombinant protein antigens of Mycobacterium tuberculosis

Recombinant β-galactosidase fusion proteins from Mycobacterium tuberculosis were purified by affinity chromatography and used for stimulation of unselected T lymphocytes freshly isolated from healthy individuals. It was found that the 12kDa, 19-kDa, 65-kDa and 71-kDa recombinant (r) proteins tested stimulated T cells from healthy individuals in an antigen-specific way. These data demonstrate that it is feasible to screen purified r-proteins with unselected T cell populations. The existence of T cells with reactivity to these antigens in healthy individuals suggests T cell activation independent from clinical disease and hence that these proteins are not indicative for active tuberculosis.     

结核分枝杆菌抗原分析及免疫交叉反应研究

目的：筛选和鉴定结核分枝杆菌特异性和保护性抗原，研究结核杆菌的免疫反应特点，以探索结核病诊断和治疗的新途径。方法：采用超声破碎和滤膜抽滤的方法分别得到菌体蛋白和滤液蛋白，通过Western blot试验用结核杆菌的单克隆抗体及结核病人血清来检测蛋白样品，把发生阳性反应的蛋白在BECKMAN LF3200／多肽氨基酸序列测定仪上进行N末端序列分析，并用结核杆菌的单抗对自身抗原组蛋白进行了检测。结果：结核杆菌的31kD和30kD蛋白与结核杆菌的单抗及病人血清反应均呈阳性，但与正常小鼠血清和健康人血清反应呈阴性。31kD和30kD蛋白的N末端序列分别为：Ala Glu Val Asp Trp Leu Val Phe Ala Val和Phe Ser Arg Pro Gly Leu Pro Val Glu Tyr。结核分枝杆菌的单抗与自身抗原组蛋白能发生免疫交叉反应。结论：结核杆菌的31kD和30kD蛋白是免疫保护性抗原，对免疫交叉反应分子基础的进一步研究必将增加对结核免疫机理的了解。