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1.
Cytokine production in synovial tissue of mice with collagen-induced arthritis (CIA) 总被引:6,自引:1,他引:6
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MÜSSENER M J LITTON E LINDROOS L KLARESKOG 《Clinical and experimental immunology》1997,107(3):485-493
The kinetics of cytokine production in arthritic limbs of mice with CIA was determined by using modified immunohistochemical techniques. Tissue cryostat sections of undecalcified whole paws were analysed for the presence of tumour necrosis factor-alpha (TNF-α), IL-6, IL-2, IL-4, IL-5 interferon-gamma (IFN-γ), transforming growth factor-beta 2 (TGF-β2) and TGF-β3. Locally produced TNF-α, IL-6 and TGF-β2 were observed within the lining layer, sublining and pannus at all stages of disease. The staining of TNF-α was particularly intense at the cartilage–pannus junction. In contrast to the monokines, IFN-γ and TGF-β3 were only expressed in scattered cells within the deeper layers of the synovia. Interestingly, IFN-γ was not present in the late phase of CIA, despite the continued presence of TNF-α and IL-6 in the pannus. Production of IL-2, IL-4 or IL-5 was not detected in any joint. The observed pattern of a relative paucity of T cell-derived cytokines and an abundance of monokines during the late phase of T cell-dependent CIA indicates that the synovial cytokine pattern previously described in rheumatoid arthritis (RA) is fully compatible with a pathogenic role of T cells. The temporal as well as spatial dissociation between expression of T cell-derived cytokines and monokines indicates that T cell-independent mechanisms may also be of importance in the triggering of monokine production during arthritis. 相似文献
2.
Autoimmune arthritis induces paired immunoglobulin‐like receptor B expression on CD4+ T cells from SKG mice
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Kathrin Rothe Nora Raulien Gabriele Köhler Matthias Pierer Dagmar Quandt Ulf Wagner 《European journal of immunology》2017,47(9):1457-1467
The chronic, destructive autoimmune arthritis in SKG mice, which closely resembles human rheumatoid arthritis, is the result of self‐reactive T cells escaping thymic deletion. Since the inhibitory receptor LIR‐1 is up‐regulated on auto‐reactive T cells in human rheumatoid arthritis, the role of its murine ortholog PIR‐B was investigated. Peripheral CD4+ T cells from SKG mice were found to frequently express PIR‐B, and this population produces more frequently IL‐17 upon in vitro stimulation compared to PIR‐B? cells. A much larger fraction of PIR‐B+ T cells, however, was found to secret no IL‐17, but IFN‐γ. With regards to the clinical course of the disease, high frequencies of PIR‐B+ CD4+ T cells were found to be associated with a milder course of arthritis, suggesting that the net effect of PIR‐B expression is suppression of autoreactive T cells. Our results indicate that overexpression of PIR‐B on IL‐17‐producing SKG CD4+ T cells might represent an effective counter‐regulatory mechanism against the destructive potential of those cells. More importantly, a major population of PIR‐B+ T cells in SKG mice appears to play an inhibitory role by way of their IFN‐γ production, since high frequencies of those cells ameliorate the disease. 相似文献
3.
D. Almeida González A. Cabrera de León C. Vázquez Moncholi B. Brito Díaz M. C. Rodríguez Pérez A. Aguirre-Jaime S. Domínguez Coello A. González Hernández 《Inflammation research》2008,57(6):266-271
OBJECTIVE: To identify the differences in cytokine profile between allogeneic and syngeneic pregnancy in mice with collagen-induced arthritis (CIA). METHODS: Mice (strain B10.RIII) were injected with bovine collagen. Females were mated with males of the same strain (syngeneic pregnancy) or with males of strain B10. Q (allogeneic pregnancy). Concentrations of cytokines were measured during pregnancy and after delivery, and the onset and evolution of arthritis was followed in all female animals throughout the study period. RESULTS: In female mice that developed CIA, cytokine concentrations were lower in allogeneic pregnancies than syngeneic pregnancies. When paired cytokine concentrations were compared in each animal during and after pregnancy, MCP-1 was lower during gestation than after delivery in both groups of pregnant mice, IL-6 was lower during gestation than after delivery only in allogeneic pregnancies, and IL-10 was lower during gestation than after delivery in allogeneic pregnancies, whereas in syngeneic pregnancies IL-10 was higher during gestation than after delivery. CONCLUSIONS: Allogeneic pregnancy was associated with less arthritis because of lower concentrations of proinflammatory cytokines (IL-6 and others), not because of an increase in the concentration of antiinflammatory cytokines (IL-10). 相似文献
4.
Dylan Daniel Ronald G. Gill Nanette Schloot Dale Wegmann 《European journal of immunology》1995,25(4):1056-1062
T cells are known to play an important role in beta cell destruction in the nonobese diabetic (NOD) mouse model of Type I diabetes and islet-specific T cell clones have been demonstrated to be capable of adoptive transfer of diabetes. One important issue involves the identity of beta cell antigens that are recognized by nominally islet cell-specific T cell clones. We have previously reported that insulin-specific T cells are a predominant component of islet-specific T cells isolated from islet infiltrates of pre-diabetic NOD mice. In this report we examine six independently derived insulin-specific T cell clones established from islet infiltrates of pre-diabetic NOD mice in detail. All six clones were found to be specific to a region of the insulin molecule defined by a synthetic peptide encompassing residues 9–23 of the B chain. Despite this restricted specificity, each member of this panel exhibited a distinct receptor specificity defined either by Vβ usage or antigen fine specificity. Five clones produced interferon (IFN)-γ but not interleukin (IL)-4, placing them in the T helper type 1 (TH1)-like category whereas one clone produced both IL-4 and IFN-γ, a characteristic of TH0 cells. All six clones were capable of either acceleration of diabetes in young NOD mice or adoptive transfer to NODscid mice. Taken together, these results suggest that spontaneously arising insulin-specific T cells participate in beta cell destruction during development of diabetes in NOD mice. 相似文献
5.
Simultaneous analysis of T cell clonality and cytokine production in rheumatoid arthritis using three-colour flow cytometry 总被引:3,自引:0,他引:3
Bakakos P Pickard C Wong WM Ayre KR Madden J Frew AJ Hodges E Cawley MI Smith JL 《Clinical and experimental immunology》2002,129(2):370-378
In this study we examined the cytokine production by T cells and TCRVbeta subsets in peripheral blood (PB) and synovial fluid (SF) from six RA patients and PB from 10 normal subjects, using three-colour flow cytometry. In two RA subjects we assessed T cell clonality by RT PCR using TCRBV family-specific primers and analysed the CDR3 (complementarity determining region 3) length by GeneScan analysis. A high percentage of IFN-gamma- and IL-2- producing cells was observed among the PB T cells in both the RA patients and normal controls and among the SF T cells in RA patients. In contrast, the percentage of T cells producing IL-4 and IL-5 was small among PB T cells in both RA patients and normal controls and among SF T cells in RA patients. There was no significant difference in the production of IFN-gamma, IL-2 and IL-5 between the two compartments (PB and SF); however, there were significantly more IL-4-producing cells in SF. Molecular analysis revealed clonal expansions of four TCRBV families in SF of two of the RA patients studied: TCRBV6.7, TCRBV13.1 and TCRBV22 in one and TCRBV6.7, TCRBV21.3 and TCRBV22 in the second. These expansions demonstrated cytokine expression profiles that differed from total CD3+ cells, implying that T cell subsets bearing various TCR-Vbeta families may have the potential to modulate the immune response in RA patients. 相似文献
6.
目的 研究盐酸小檗碱(BBR)对小鼠脾细胞增殖与凋亡及细胞因子产生的影响.方法 采用无菌取BALB/c小鼠脾脏,制成脾细胞悬液,脾细胞预先用(1、2、4) μg/mL BBR处理60 min,加入刀豆蛋白(ConA)刺激脾细胞增殖,细胞培养至24、48、72 h时,用MTT方法检测脾细胞增殖情况;流式细胞术检测脾细胞培养不同时间的凋亡情况,用细胞因子ELISA检测试剂盒定量分析细胞上清液中TNF-α,IL-2和IFN-γ的浓度.结果 与对照组相比,在以上浓度范围内,BBR对ConA刺激下实验组小鼠脾细胞的增殖和TNF-α,IL-2,IFN-γ产生有明显的抑制作用(P<0.05),且呈浓度及时间依赖性,但对小鼠脾细胞凋亡无明显影响(P>0.05).结论 BBR对小鼠脾细胞具有明显的免疫抑制作用,可作为潜在的免疫抑制药物. 相似文献
7.
Cytokine profiles of BAL T cells and T-cell clones obtained from human asthmatic airways after local allergen challenge. 总被引:8,自引:0,他引:8
K J Bodey A E Semper A E Redington J Madden L M Teran S T Holgate A J Frew 《Allergy》1999,54(10):1083-1093
BACKGROUND: This study assessed the heterogeneity of cytokine expression in asthma before and after local allergen challenge. METHODS: BAL T cells were obtained 10 min or 24 h after local endobronchial allergen challenge in atopic asthmatic subjects. T cells were cloned by direct limiting dilution. mRNA expression was assessed by RT-PCR, and cytokine protein production by ELISA. RESULTS: Unstimulated baseline BAL T cells expressed mRNA for IFN-gamma, IL-13, and TNF-alpha. A minority of samples expressed IL-4 and IL-5, but no IL-3 mRNA was detected. PHA stimulation increased expression of IL-3, IL-4, and IL-5 mRNA in 4/6 samples. IL-13 and GM-CSF mRNA were found in BAL cells after allergen challenge, but expression of IFN-gamma was reduced. Both IL-4 and IL-3 were strongly upregulated after PHA stimulation, while the expression of TNF-alpha and IFN-gamma was reduced, compared to equivalent baseline samples. Seventeen panels of BAL T-cell clones were derived (average cloning efficiency 1/40 T cells). Seven panels survived to 8 weeks for analysis. Clones derived 4 h after saline challenge showed strong mRNA signals for IL-13, IL-4, and IFN-gamma, whereas clones derived 24 h after allergen challenge expressed IL-13, GM-CSF, IL-3, IL-4, and often IL-5 (i.e., closer to the Th2 profile). There was considerable heterogeneity in the patterns of cytokine mRNA and protein production by different clones. CONCLUSIONS: T cells from asthmatic airways produce IL-13, IFN-gamma, and TNF-alpha, but after allergen challenge, type 2 cytokines are upregulated. mRNA and protein analysis provide complementary information on airways T-cell cytokine profiles. 相似文献
8.
类风湿关节炎小鼠关节聚集的T细胞克隆型变化的分析 总被引:5,自引:2,他引:3
目的 :通过对类风湿性关节炎 (RA)小鼠模型发病不同时期的四肢足关节聚集的T细胞克隆型的分析 ,了解T细胞克隆型与RA病变的关系。方法 :用RT PCR/SSCP法 ,对一种自发性RA小鼠模型 (SKG小鼠 )发病初期与后期四肢足关节聚集的T细胞克隆型进行比较分析。结果 :在发病后期 ,四肢足关节中一致的Vβ2和Vβ8.2T细胞克隆型均明显升高 (分别为 72 .3%和 6 0 .2 % )。结论 :TCR为Vβ2和Vβ8.2的T细胞克隆 ,可能对于SKG小鼠RA的发展起重要作用 相似文献
9.
C. Contigli D. N. Silva-Teixeira G. Del Prete M. M. D'Elios M. De Carli M. Manghetti A. Amedei F. Almerigogna J. R. Lambertucci A. M. Goes 《Clinical immunology (Orlando, Fla.)》1999,91(3):338-344
It is essential to distinguish the role of T lymphocytes on the physiopathology associated to more severe forms of schistosomiasis and on the immunomodulation that evolves in the majority of infected people. In this study, we generated Schistosoma mansoni-specific T cell lines and clones from patients with the acute and chronic (intestinal and hepatosplenic forms) phases of disease, from former ones, and from uninfected individuals sensitized to parasite soluble antigens. T cell lines derived from nontreated acute infected donors were capable of producing IL-4 and IL-5, while cells from treated patients secreted IFN-gamma. Lines from intestinal chronic and antigen-sensitized donors preferentially produced IFN-gamma, while those from hepatosplenic patients secreted all three cytokines. The cytokine analysis of CD4+ T cell clones revealed a Th2/Th0 pattern (clones producing IL-4 and IL-5 and clones producing all three cytokines) for those derived from infected patients, while cells from antigen-sensitized donors exhibited an opposite Th1/Th0 pattern (clones producing IFN-gamma and clones producing all three cytokines). The possible role of these T cell populations on human schistosomiasis mansoni is discussed. 相似文献
10.
背景:骨髓间充质干细胞的多向分化能力在骨代谢疾病中发挥重要作用,受激素、细胞因子等多种因素调节。目前骨髓间充质干细胞骨向分化的表观遗传学调控机制尚不明确,组蛋白去乙酰化酶与骨质疏松的关系尚需进一步探讨。
目的:建立雌激素缺乏骨质疏松小鼠的实验动物模型,检测骨髓间充质干细胞组蛋白去乙酰化酶1,3,4 mRNA表达水平,探索卵巢切除小鼠骨组织形成障碍的表观遗传学机制。
方法:昆明种小鼠30只随机等分为模型组和假手术组。小鼠适应性喂养7 d后,模型组小鼠切除双侧卵巢,造成雌激素缺乏骨质疏松实验动物模型,假手术组小鼠仅切除等量脂肪组织。
结果与结论:模型组小鼠股骨骨小梁稀疏或断裂,骨小梁宽度变窄,骨小梁间距变宽,骨小梁占视野面积降低。与假手术组相比,模型组小鼠骨髓间充质干细胞中组蛋白去乙酰化酶3 mRNA表达水平显著降低,组蛋白去乙酰化酶1,4表达水平变化差异无显著性意义。提示雌激素缺乏导致骨髓间充质干细胞去乙酰化状态改变可能是骨形成障碍的重要原因之一。中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接: 相似文献
11.
目的 用全血在单个细胞水平上研究类风湿性关节炎 (RA)患者TH1 TH2 细胞因子分泌模式及其与疾病活动程度的关系。方法 用三色流式细胞术检测RA患者外周血细胞内细胞因子IFN γ和IL 4的表达情况。结果 RA患者外周血IFN γ 细胞百分率及IFN γ /IL 4 细胞比值比健康人明显增高 ,IFN γ /IL 4 比值与患者Stoke指数呈正相关 (r=0 .86 10 ,P <0 .0 1)。结论 RA患者外周血中细胞因子分泌模式朝TH1 偏移 ,IFN γ /IL 4 比值与疾病活动性相关 ,TH1 TH2 平衡在RA发病机理及疾病进程中起重要作用 相似文献
12.
Porphyromonas gingivalis infection exacerbates the onset of rheumatoid arthritis in SKG mice
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M. Yamakawa K. Ouhara M. Kajiya S. Munenaga M. Kittaka S. Yamasaki K. Takeda K. Takeshita N. Mizuno T. Fujita E. Sugiyama H. Kurihara 《Clinical and experimental immunology》2016,186(2):177-189
Epidemiological studies have linked periodontitis to rheumatoid arthritis (RA). Porphyromonas gingivalis (Pg) was reported recently to produce citrullinated protein (CP) and increase anti‐cyclic CP antibody (ACPA), both of which have been identified as causative factors of RA. In the present study, we determined the effects of Pg infection on the exacerbation of RA in a mouse model. RA model mice (SKG mice) were established by an intraperitoneal (i.p.) injection of laminarin (LA). Mice were divided into six groups, Ctrl (PBS injection), LA (LA injection), Pg/LA (Pg + LA injection), Pg (Pg injection), Ec/LA (Escherichia coli and LA injection) and Ec (E. coli injection). In order to evaluate RA, joint swelling by the arthritis score, bone morphology by microcomputed tomography (microCT), haematoxylin and eosin staining, ACPA, matrix metalloproteinase‐3 (MMP‐3) and cytokine level in serum by enzyme‐linked immunosorbent assay were determined. Osteoclast differentiation from bone marrow mononuclear cells (BMCs) was examined to clarify the underlying mechanisms of RA. The presence of Pg and CP in joint tissue was also investigated. The arthritis score was threefold higher in the Pg/LA group than in the LA group. Severe bone destruction was observed in joint tissue of the Pg/LA group. A microCT analysis of the Pg/LA group revealed a decrease in bone density. ACPA, MMP‐3, interleukin (IL)‐2, IL‐6, CXCL1 and macrophage inflammatory protein (MIP)‐1α levels from the Pg/LA group were the highest. The osteoclastogenesis of BMCs was enhanced in the Pg/LA group. Furthermore, large amounts of Pg components and CP were detected in the Pg/LA group. In conclusion, Pg infection has the potential to exacerbate RA. 相似文献
13.
Allergen-stimulated T lymphocytes from allergic patients induce vascular cell adhesion molecule-1 (VCAM-1) expression and IL-6 production by endothelial cells. 总被引:1,自引:1,他引:1
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Y Delneste P Jeannin P Gosset P Lassalle E Cardot I Tillie-Leblond M Joseph J Pestel A B Tonnel 《Clinical and experimental immunology》1995,101(1):164-171
Adhesion of inflammatory cells to endothelium is a critical step for their transvascular migration to inflammatory sites. To evaluate the relationship between T lymphocytes (TL) and vascular endothelium, supernatants from allergen-stimulated TL obtained from patients sensitive to Dermatophagoides pteronyssinus (Dpt) versus healthy subjects were added to endothelial cell (EC) cultures. TL were stimulated by autologous-activated antigen-presenting cells (APC) previously fixed in paraformaldehyde to prevent monokine secretion. Two parameters were measured: the expression of adhesion molecule and the production of IL-6. Related allergen-stimulated TL supernatants from allergic patients induced an increase of VCAM-1 and intercellular adhesion molecule-1 (ICAM-1) expression when supernatants of the control groups (TL exposed to an unrelated allergen or not stimulated or TL obtained from healthy subjects) did not. E-selectin expression was not modulated whatever the supernatant added to EC culture. IL-6 production by EC was significantly enhanced after activation with related allergen-stimulated TL supernatants from allergics compared with control supernatants. Induction of VCAM-1 expression was inhibited by adding neutralizing antibodies against IL-4, whereas IL-6 production and ICAM-1 expression were inhibited by anti-interferon-gamma (IFN-gamma) antibodies. Enhanced production of IL-4 and IFN-gamma was detected in related allergen-stimulated TL supernatants from allergic subjects compared with the different supernatants. These data suggest that allergen-specific TL present in the peripheral blood of allergic patients are of Th1 and Th2 subtypes. Their stimulation in allergic patients may lead to the activation of endothelial cells and thereby participate in leucocyte recruitment towards the inflammatory site. 相似文献
14.
《Immunological investigations》2013,42(2):144-156
Members of the Mycobacterium tuberculosis (Mtb) Beijing genotype are a major concern due to their high prevalence in tuberculosis patients and their high rate of multi-drug resistance. Although it has been shown that Beijing modifies macrophage behavior, little is known about how this genotype could affect the cellular immune response. In order to address this issue, peripheral blood mononuclear cells (PBMC) from healthy BCG vaccinated individuals were stimulated with protein extracts from three Mycobacterium tuberculosis genotypes: Canetti, H37Rv and Beijing evaluating T cell proliferation and cytokine production. In this system both CD4+ and CD8+ proliferated in a similar manner independently of the Mtb genotype used for stimulation. Regarding cytokines, all strains induced similar levels of IFN-γ, but were unable to induce IL-4 and TGF-β. Contrasting, Canetti strain induced lower production of IL-10, TNF-α and IL-12 compared to H37Rv and Beijing. Interestingly, PBMC stimulated with the Beijing strain produced the highest levels of IL-12 and IL-10 than those stimulated with other strains. This differential cytokine expression could affect the pathogenesis induced by Beijing strain through the modulation of inflammatory process in the host, but the precise mechanisms by which this cytokine environment affects the Beijing strain pathogenesis needs further characterization. 相似文献
15.
16.
Son A Nakamura H Okuyama H Oka S Yoshihara E Liu W Matsuo Y Kondo N Masutani H Ishii Y Iyoda T Inaba K Yodoi J 《European journal of immunology》2008,38(5):1358-1367
Thioredoxin-binding protein-2 (TBP-2), also known as vitamin D3-up-regulated protein 1 (VDUP1), was identified as an endogenous molecule interacting with thioredoxin (TRX). Here, we show that dendritic cells (DC) derived from TBP-2-deficient mice are defective in the function of T cell activation. To compare TBP-2(-/-) DC function with wild-type (WT) DC, we stimulated DC with lipopolysaccharide (LPS). Although TBP-2(-/-) DC and WT DC expressed comparable levels of MHC class II and costimulatory molecules such as CD40, CD80 and CD86, the IL-12p40, IL-12p70 and IL-6 productions of TBP-2(-/-) DC were attenuated. In a mixed leukocyte reaction (MLR), the concentrations of IL-2, IFN-gamma, IL-4 and IL-10 in the culture supernatant of MLR with TBP-2(-/-) DC were significantly lower than those in the cultures with WT DC. In MLR also, as with LPS stimulation, IL-12p40 and IL-12p70 production from TBP-2(-/-) DC was less than that from WT DC. Proliferation of T cells cultured with TBP-2(-/-) DC was poorer than that with WT DC. In vivo delayed-type hypersensitivity responses in TBP-2(-/-) mice immunized with ovalbumin were significantly reduced compared to WT mice. These results indicate that TBP-2 plays a crucial role in DC to induce T cell responses. 相似文献
17.
结核胸液中ESAT-6多肽特异性多功能CD4+T细胞数量及功能分析 总被引:1,自引:0,他引:1
目的:检测ESAT-6多肽刺激后,结核性胸膜炎患者胸液细胞中CD4+T细胞细胞因子产生及多功能CD4+T细胞频率,了解ESAT-6特异性CD4+T细胞在结核局部细胞免疫应答中的作用.方法:分离结核性胸膜炎患者胸液细胞(PFCs),ESAT-6混合多肽刺激后检测CD4+T细胞细胞因子分泌、细胞亚群、多功能性CD4+T细胞频率及细胞因子平均荧光强度.结果:BCG、ESAT-6混合多肽和ESAT-6组蛋白刺激PFCs后,主要是CD4+T细胞分泌Th1细胞因子(IFN-γ、IL-2和TNF-α),而CD8+T细胞几乎不产生细胞因子.进一步分析ESAT-6混合多肽刺激PFCs后Th1细胞的亚群组成,依据细胞因子分泌的类型及数量,该特异性Th1细胞可以分为7个不同亚群,且各亚群所占比例不同,其中多功能性T细胞亚群(同时分泌IFN-γ、IL-2和TNF-α)比例较高.细胞因子荧光强度分析表明,依据细胞因子分泌类型的增加,单个细胞水平上不同Th1亚群中各细胞因子表达的量也逐渐增加,即3+>2+>1+细胞.结论:ESAT-6混合多肽刺激结核性胸膜炎患者胸液细胞后,主要诱导CD4+T细胞分泌细胞因子,且Th1亚群中包含多功能性CD4+T细胞,该细胞在单细胞水平上分泌细胞因子的类型和数量也显著高于其他亚群.可能在结核局部感染中发挥关键保护性作用. 相似文献
18.
The effects of mesoporphyrin, a novel porphyrin derivative, on type II collagen-induced arthritis in mice were studied. Mesoporphyrin (10–30 mg/kg) and prednisolone (5 mg/kg; reference drug) reduced the incidence and severity of type II collagen-induced arthritis in mice, as assayed by clinical observation and histopathological studies. Although both agents inhibited type II collagen-induced delayed type hypersensitivity in arthritic mice, only prednisolone inhibited humoral immunity to type II collagen. The effects of mesoporphyrin on T cell dependent allergic inflammation were examined, in order to study the mechanism by which it inhibits arthritis. Staphylococcal enterotoxin B (SEB; superantigen)-potentiated collagen-induced arthritis and sheep red blood cell-induced delayed type hypersensitivity reaction were clearly inhibited by mesoporphyrin. Moreover, the superantigen-induced CD-25 expression on T cells was inhibited by mesoporphyrin. These results indicate that mesoporphyrin inhibits type II collagen-induced arthritis by inhibiting the activation of T cells.accepted by M. Katori 相似文献
19.
不同刺激剂和培养环境对CD4+、CD8+T细胞内细胞因子表达的影响 总被引:2,自引:1,他引:2
目的探讨不同的刺激剂和不同的培养条件对CD4 和CD8 T细胞内细胞因子表达的影响。方法分离正常人的外周血单个核细胞(PBMC),分别加入3种不同的刺激剂(PHA,抗CD3和抗CD28mAb,PMA和离子霉素),置于4种不同的培养环境下(室温,37℃水浴,37℃培养箱,37℃50mL/LCO2培养箱)培养4·5~5h。收集细胞,以荧光素-mAb标记后,用流式细胞术分析CD4 、CD8 T细胞内IL-2、IFN-γ和TNF-α的表达。结果CD4 和CD8 T细胞内细胞因子的表达,随着刺激剂的不同而有所差别,且在上述4种培养环境中,PMA的刺激效果最强,抗CD3mAb次之,PHA的刺激效应最弱。以上述3种刺激剂刺激后,不同培养条件对T细胞内细胞因子的表达有一定的影响。室温培养时几乎检测不到细胞因子的表达,而在37℃水浴、37℃培养箱和37℃50mL/LCO2培养箱中培养时,表达细胞因子的CD4 和CD8 T细胞的百分率差异无统计学意义(P>0.05)。结论不同刺激剂体外刺激T细胞表达细胞因子的效应不同,依次为PMA和离子霉素>抗CD3和抗CD28mAb>PHA。体外刺激培养的T细胞活化过程中,温度是重要的条件,CO2无明显影响。 相似文献