Glucocorticoid resistance in chronic asthma. Glucocorticoid pharmacokinetics, glucocorticoid receptor characteristics, and inhibition of peripheral blood T cell proliferation by glucocorticoids in vitro

A total of 37 chronic, severe, nonsmoking asthmatic patients with documented reversible airways obstruction were classified as glucocorticoid-sensitive or -resistant on the basis of changes in FEV1, FVC, and peak expiratory flow (PEF) after oral prednisolone. The resistant patients showed no significant improvements in airflow limitation. Phytohemagglutinin (PHA)-induced proliferation of peripheral blood T lymphocytes from the sensitive but not the resistant asthmatic patients was significantly (p less than 0.01) inhibited by dexamethasone (10(-7) mol/L), reflecting a shift of the dose-response curve. When all the asthmatic patients were analyzed together, there was a significant correlation between the degree of sensitivity of T cells to dexamethasone and the clinical responsiveness to prednisolone (p less than 0.01). No differences were observed between six of the sensitive and resistant patients in the clearance of plasma prednisolone derived from orally administered prednisone. Peripheral blood mononuclear cell glucocorticoid receptors were also characterized in five sensitive and seven resistant patients. The numbers and binding affinities of these receptors could not account for the observed difference in the susceptibility of these cells to functional inhibition by dexamethasone in vitro. These results suggest that clinical glucocorticoid resistance in chronic asthma does not reflect abnormal glucocorticoid clearance but may be due at least partly to a relative insensitivity of T lymphocytes to glucocorticoids. This lack of sensitivity is unexplained but is not attributable to abnormalities of cellular glucocorticoid receptors.     

In vitro production of myeloperoxidase anti-neutrophil cytoplasmic antibody and establishment of Th1-type T cell lines from peripheral blood lymphocytes of patients

OBJECTIVE: To investigate the pathogenic role of T cells in the development of anti-neutrophil cytoplasmic antibody (ANCA) associated vasculitis. METHODS: Peripheral blood lymphocytes (PBL) were isolated from myeloperoxidase anti-neutrophil cytoplasmic antibody (MPO-ANCA) associated vasculitis patients and cultured in medium. The production of MPO-ANCA in the medium of PBL stimulated with Concanavalin-A (Con-A), with or without cyclosporin (CyA), was measured by enzyme-linked immunosorbent assay (ELISA) on MPO coated plates. RNA isolated from PBMC of one patient was used for polymerase chain reaction (PCR) and single stranded conformational polymorphism (SSCP) studies, and MPO-specific T cell lines (TCL) were established by antigen stimulation techniques. RESULTS: PBL of patients with MPO-ANCA-associated vasculitis produced MPO-ANCA following Con-A stimulation, and this effect was inhibited by treatment with cyclosporin A (CyA) or elimination of CD4 cells. PCR-SSCP showed autoantigen-reactive oligoclonal T-cell accumulation in PBMC of one of these patients. We established MPO-specific TCL which secreted interferon-gamma (IFN-gamma), but not interleukin-4 (IL-4); all TCL were CD4 positive, CD8 negative, and HLA-DR restricted. CONCLUSIONS: Our results suggest that Th1-type T cells may mediate MPO-ANCA production, and may play a role in the onset of MPO-ANCA vasculitis.     

结核病小鼠T淋巴细胞亚群及其表达的四项细胞因子分析

目的 探讨T淋巴细胞(简称T细胞)表达穿孔素(PFN)、颗粒酶B(GzmB)、γ-干扰素(IFN-γ)、白细胞介素-2(IL-2)与结核免疫的关系.方法 60只MK小鼠按随机数字表法分成结核病组和健康对照组,每组30只.以CD3PerCP、CD4FTTC、CD8APC单抗标记T细胞亚群,以藻红蛋白标记PFN、GzmB、IFN-γ、IL-2单抗标记细胞因子,以流式细胞仪检测分析总的淋巴细胞、CD3+、CD4+、CD8+、CD4+CD8+;双阳(DP)T细胞计数和各亚群占淋巴细胞百分率,观察各T细胞亚群内表达PFN、GzmB、IFN-γ、IL-2的阳性细胞计数和各自占淋巴细胞百分率.采用t检验进行统计学分析.结果 (1)CD4+CD8+、DP T细胞均能不同程度地表达PFN、GzmB、IFN-γ、IL-2.PFN、GzmB的表达以CD8+T细胞占优势.IFN-γ、IL-2的表达以CD4+T细胞占优势.(2)T细胞亚群细胞计数结果与T细胞亚群占总淋巴细胞百分率结果,所反映的T细胞免疫变化趋势可能相似,也可能不同甚至相反.(3)两组间表达PFN的T细胞差别不明显,但结核病组表达GzmB的各个T细胞亚群计数和CD3+%、CD8+%、DP%高于对照组(t值为-3.72～4.13.均P<0.05).(4)结核病组表达IFN-γ的CD3+、CD4+T细胞计数高于对照组;但结核病组表达IL-2的CD8+、DP T细胞计数和CD3+%、CD4+%、CD8+%、DP%均低于对照组(t值为2.62～3.46,均P<0.05).结论 、CD,4+、CD8+、DPT细胞均能不同程度地表达PFN、GzmB、IFN-γ、IL-2;联合评价T细胞各亚群计数和其占淋巴细胞百分率更能判断免疫学状态.     

Apoptosis and B-cell lymphoma-2 of peripheral blood T lymphocytes and soluble fas in patients with allergic asthma

STUDY OBJECTIVE:s: The dysregulation of apoptosis and the expression of apoptosis-related molecules of allergen-reactive T lymphocytes have been suggested to play a key role in the development and maintenance of the inflammatory reactions in allergic asthma. Glucocorticoids are effective drugs for treating allergic inflammation. In this study, we investigated the effect of dexamethasone (DEX) on the apoptosis and B-cell lymphoma (Bcl)-2 expression of peripheral blood T lymphocytes as well as the soluble form of Fas (sFas) in allergic asthmatic patients. METHODS: Peripheral blood lymphocytes from 41 allergic asthmatic patients and 30 age-matched and sex-matched control subjects were treated with 0.1 and 1 micro M DEX. The percentages of apoptosis and expression of the Bcl-2 molecule in T lymphocytes were assessed by flow cytometry. The plasma concentration of sFas was measured using the enzyme-linked immunosorbent assay. RESULTS: DEX (0.1 and 1 micro M) could induce the apoptosis of T lymphocytes from allergic asthmatic patients and control subjects in a dose-dependent manner in vitro. The apoptotic susceptibility of T lymphocytes to DEX and the plasma sFas concentration were significantly higher in allergic asthmatics. The ex vivo expression of Bcl-2 was significantly lower in the T lymphocytes of asthmatic patients than in those of the control subjects. However, DEX did not have any significant effect on the expression of Bcl-2 in vitro. CONCLUSIONS: The T lymphocytes of asthmatic patients have higher apoptotic susceptibility to DEX treatment in vitro and a lower expression of the Bcl-2 molecule.     

T lymphocyte response to Neisseria gonorrhoeae porin in individuals with mucosal gonococcal infections.

T lymphocytes from a majority of patients with urogenital gonococcal disease (67%-80%) proliferated on incubation with gonococcal porin (Por), compared with minimal induced proliferation of T lymphocytes from normal volunteers. A significant increase in Por-specific interleukin (IL)-4-producing CD4+ T helper lymphocytes was seen in patients with mucosal gonococcal disease and not in normal controls. Similar results were observed in CD8+ T lymphocytes from these patients. There was no measured increase in IL-2, IL-10, IL-12, interferon-gamma, or tumor necrosis factor-alpha production by T lymphocytes from infected subjects on incubation with Por. Concomitant increases in IL-4 production in T lymphocytes from infected subjects expressing the mucosal addresin VLAalpha4/beta7 on their surface were also observed on Por incubation, but the increases were similar in T lymphocytes that were VLAalpha4/beta7 negative. In conclusion, mucosal gonococcal disease can induce Por-specific circulating T lymphocytes with a Th2 phenotype, and a portion of these Por-specific T lymphocytes can potentially traffic to mucosal surfaces.     

CD4+NKG2D+ T cells in Crohn's disease mediate inflammatory and cytotoxic responses through MICA interactions

BACKGROUND & AIMS: Crohn's disease (CD) is an inflammatory bowel disease characterized by uncontrolled immune responses to bacterial flora, with excessive activation of T lymphocytes. MICA is a stress-induced major histocompatibility complex-related molecule expressed on normal intestinal epithelial cells (IECs) and recognized by the NKG2D-activating receptor on CD8(+) T cells, gammadelta T cells, and natural killer cells. We examined the role of MICA-NKG2D interactions in the activation of T lymphocytes in CD. METHODS: MICA expression was analyzed by flow cytometry on IECs isolated from patients with active inflammatory bowel disease and controls. NKG2D expression and function were analyzed on lamina propria and peripheral blood lymphocytes. RESULTS: MICA expression was significantly increased on IECs in CD, with higher expression in macroscopically involved areas. A subset of CD4(+) T cells expressing NKG2D was increased in the lamina propria from patients with CD compared with controls and patients with ulcerative colitis. CD4(+)NKG2D(+) T cells with a Th1 cytokine profile and expressing perforin were increased in the periphery and in the mucosa in CD. CD4(+)NKG2D(+) T-cell clones were functionally active through MICA-NKG2D interactions, producing interferon-gamma and killing targets expressing MICA. IECs from patients with CD had the ability to expand this subset in vitro. CD4(+)NKG2D(+) lamina propria lymphocytes from patients with CD highly expressed interleukin-15R alpha, and interleukin-15 increased NKG2D and DAP10 expression in CD4(+)NKG2D(+) T-cell clones. CONCLUSIONS: These findings highlight the role of MICA-NKG2D in the activation of a unique subset of CD4(+) T cells with inflammatory and cytotoxic properties in CD.     

Interferon-gamma marks activated T lymphocytes in AIDS patients

Lymphocytes expressing interferon-gamma (IFN-gamma) on their surface were evaluated in 61 patients, all IV drug abusers, infected with human immunodeficiency virus type 1 (HIV-1), and in 85 healthy subjects (61 of whom were blood donors and 24 HIV-1 seronegative IV drug abusers). Data obtained demonstrated that IFN-gamma-expressing T lymphocytes, mostly CD8+ cells, were present in HIV-1-infected patients, and that their percentage, always higher in HIV-1-infected patients than in healthy subjects (p less than or equal to 0.001), increased with progressive stages of HIV-1 infection. At the same time other markers of T-cell activation, namely interleukin-2 receptor (rIL-2), transferrin receptor, and HLA-DR were also found to be positive in some of the HIV-1-infected subjects. The presence in the HIV-1-infected patients of activated CD8+ T cells, which are resistant to HIV-1 infection, may suggest that these cells are able to respond to continuous and progressive viral expression (HIV or/and other viruses) and may be a component of the specific response to HIV-1.     

Gamma delta T lymphocytes in human tuberculosis.

The manifestations of tuberculous infection reflect the immune response to infection. Most healthy tuberculin reactors develop protective immunity; tuberculous pleuritis reflects a resistant response manifest by mild disease, whereas advanced pulmonary and miliary tuberculosis reflect ineffective immunity. The role of gamma delta T cells was assessed in tuberculous infection by evaluating expansion of these cells from blood mononuclear cells after stimulation with Mycobacterium tuberculosis. After culture in vitro, the percentages of gamma delta+ cells were significantly greater in patients with protective and resistant immunity (tuberculin reactors, 25% +/- 4%; tuberculous pleuritis, 30% +/- 7%) than in those with ineffective immunity (advanced pulmonary tuberculosis, 9% +/- 3%; miliary tuberculosis, 2% +/- 1%). In leprosy, expansion of gamma delta+ cells was greater in immunologically resistant tuberculoid patients (32% +/- 4%) than in Mycobacterium leprae-unresponsive lepromatous patients (9% +/- 2%). M. tuberculosis-reactive gamma delta T cell lines produced interferon-gamma, granulocyte-macrophage colony-stimulating factor, interleukin-3, and tumor necrosis factor-alpha, cytokines that activate macrophages and may contribute to mycobacterial elimination. These findings suggest that gamma delta T cells contribute to immune resistance against M. tuberculosis.     

地塞米松对支气管哮喘急性发作患者外周血T淋巴细胞亚群自噬的影响

目的 本文拟研究地塞米松对哮喘急性发作患者外周血T淋巴细胞亚群自噬的影响.方法 分离哮喘组及健康者外周血T淋巴细胞亚群(CD+4 T,CD+8 T和 CD+4CD+25 T 细胞),分别与地塞米松(10-5 mol·L-1)共培养.首先以电子显微镜及荧光显微镜观察培养后细胞的自噬形态学改变;然后丹(磺)酰戊二胺(MDC)染色后,以流式细胞术检测上述细胞的自噬水平及CD+4CD+25 T细胞的Foxp3表达.结果 ①镜下可观察到与地塞米松共培养后细胞的典型自噬形态学改变;②地塞米松可以上调哮喘组外周血CD+4 T和 CD+4CD+25 T 细胞的自噬率(P<0.05),自噬水平的升高均呈时间依赖性 (P<0.05);③哮喘组CD+4CD+25 T细胞的Foxp3表达显著低于健康对照组(P<0.05),但地塞米松预处理对Foxp3的表达无影响(P>0.05).结论 地塞米松诱导哮喘急性发作患者外周血T淋巴细胞亚群自噬水平的增高可能是糖皮质激素治疗哮喘的作用机制之一.     

T lymphocyte adhesion to human synovial fibroblasts. Role of cytokines and the interaction between intercellular adhesion molecule 1 and CD11a/CD18.

We studied the adhesion of human peripheral blood T lymphocytes to human synovial fibroblasts stimulated with interferon-gamma (IFN gamma), tumor necrosis factor alpha (TNF alpha), interleukin-1 beta (IL-1 beta), or combinations of these cytokines. T lymphocytes bound poorly to untreated human synovial fibroblasts. IFN gamma treatment resulted in the largest increase in adhesion, followed by TNF alpha and IL-1 beta. Combinations of IFN gamma + TNF alpha and IFN gamma + IL-1 beta had a synergistic effect on intercellular adhesion molecule 1 (ICAM-1) expression and adhesion. The increase in cellular adhesion induced by cytokines correlated with the up-regulation of the number of cells expressing ICAM-1 and the density of antigen/cell. There was no synergistic effect on leukocyte function-associated antigen 3 (LFA-3) or on HLA class I or class II antigen expression. Adhesion was only partially inhibited by anti-ICAM-1, anti-LFA-1, or anti-CD18. These findings suggest the existence of ICAM-1--independent and CD11/CD18-independent adhesion mechanisms. Anti-LFA-3 was completely ineffective as an inhibitor of adhesion. There was no additive or synergistic advantage of using combinations of antibodies to increase the level of inhibition, i.e., anti--ICAM-1 + anti-LFA-3, anti-ICAM-1 + anti-CD18, or anti-ICAM-1 + anti-LFA-1 (CD11a). Our data indicate that proinflammatory cytokines may play a prominent role in the formation and exacerbation of synovial hyperplasia, by regulating the recruitment and retention of T lymphocytes via the up-regulation of adhesion molecules on synovial fibroblasts.     

γδT细胞与支气管哮喘

γδT细胞是一种在肺部含量较多的T细胞亚群.在发育、分布、表面标志和抗原识别等方面,都具有不同于αβT细胞的特性.支气管哮喘(简称哮喘)个体肺部γδT细胞的比例较健康人明显升高.在哮喘发病的不同阶段,不同的γδT细胞亚群先后发挥正向或负向的免疫调节作用.使用糖皮质激素,特异性致敏原皮下脱敏疗法,接种卡介苗,吸入单克隆抗体等方法,均可以通过作用于γδT细胞来发挥防治哮喘的作用.     

Involvement of P-glycoprotein in the transmembrane transport of interleukin-2 (IL-2), IL-4, and interferon-gamma in normal human T lymphocytes

The physiological role of the multidrug resistance P-glycoprotein (P- gp), which is expressed by normal human T lymphocytes, is still largely unknown. To investigate whether or not P-gp is involved in the transport of cytokines, peripheral blood lymphocytes were stimulated with phytohemagglutinin (PHA) in the absence or presence of P-gp inhibitors, and concentrations of cytokines (interleukin-2 [IL-2], IL- 4, IL-6, interferon-gamma [IFN-gamma]) in the supernatants of these cultures were quantitated by enzyme-linked immunosorbent assay. P-gp inhibitors included verapamil (Ver), tamoxifen (Tmx), and the P-gp specific monoclonal antibody UIC2. Release of IL-2 was significantly suppressed by these inhibitors at concentrations that were also effective in blocking efflux of Rhodamine-123 from normal T lymphocytes. IL-2 mRNA expression in lymphocytes was not different between PHA control and the cultures with P-gp inhibitors. Ver and Tmx did not interfere with T-cell activation as determined by CD25 and CD69 expression. In a nonhematological model, the P-gp expressing HCT-8 adenocarcinoma cell line, exogenously added IL-2 was shown to exert an inhibitory effect on P-gp mediated Rhodamine-123 efflux. In addition, transepithelial transport of IL-2 by electrophysiologically tight and polarized HCT-8 monolayers was examined. A time-dependent flux of IL-2 across dense monolayers, which was partially inhibited by Ver, was observed. We also investigated whether or not P-gp inhibitors suppressed release of other cytokines produced by activated T cells (IL- 4, IL-6, IFN-gamma). Release of IL-4 and IFN-gamma was significantly inhibited by Ver, Tmx, and UIC2; however, release of IL-6 remained unaffected. These data show P-gp mediated transmembrane flux of IL-2 in T lymphocytes and HCT-8 cells. We conclude that P-gp participates in the transport of cytokines (IL-2, IL-4, and IFN-gamma) in normal peripheral T lymphocytes.     

中国乙型肝炎患者外周血淋巴细胞亚群频率参考值范围

目的 调查中国乙型肝炎患者外周血淋巴细胞亚群频率参考值范围.方法 利用流式细胞术检测2846例乙型肝炎患者和117例健康人群外周血淋巴细胞亚群数值,调查我国健康人群和乙型肝炎人群的参考值范围.结果 调查了16～60岁健康人群和HBV感染相关的急性肝炎、慢性肝炎、重型肝炎和肝硬化人群外周血CD3+T淋巴细胞、CD3+CD...     

急性和慢性乙型肝炎患者外周血NK细胞和T淋巴细胞亚群的动态变化

目的：观察急性和慢性乙型肝炎患者急性期和恢复期外周血NK细胞和T淋巴细胞亚群的变化。方法在40例急性乙型肝炎和40例慢性乙型肝炎患者,分别在急性期和恢复期检测CD3＋CD4＋T细胞、CD3＋CD8＋T细胞和NK（CD3-CD16＋CD56＋）细胞占淋巴细胞的比率（%）。结果在急性乙型肝炎急性期NK细胞计数为（15.7±7.5）%,而在恢复期则上升至（21.9±8.2）%,（P＜0.05）;急性乙型肝炎患者在急性期CD3＋CD4＋T细胞为（35.5±6.8）%,到恢复期则显著下降（33.6±7.0）%,（P＜0.05）;急性乙型肝炎在急性期CD3＋CD8＋T细胞为（35.6±7.6）%,而在恢复期则显著下降（30.0±7.5）%,（P＜0.05）,后者仍比慢性乙型肝炎患者在病情恢复期高（19.1±7.1）%,（P＜0.05）。结论在急性乙型肝炎病程中,NK细胞呈上升趋势,CD3＋CD8＋T细胞呈下降趋势,而在慢性乙型肝炎患者NK细胞及T淋巴细胞数量下降,致病情迁延不愈。     

哮喘患者外周血T淋巴细胞凋亡及其分子机制

目的　探讨哮喘患者外周血 Ｔ淋巴细胞凋亡率的变化及其分子机制。方法　取１２ 例急性发作期哮喘患者（ 哮喘组） 及１０ 名正常对照者（ 对照组） 外周血淋巴细胞并无菌分离 Ｔ 淋巴细胞,采用电镜和原位末端终止法（ Ｔ Ｕ Ｎ Ｅ Ｌ） 观察抗 Ｃ Ｄ＋３ 单抗（１００ ｍｇ／ Ｌ） 诱导体外培养０ 、２４ 、４８ 、７２ 小时后 Ｔ淋巴细胞凋亡变化,同时采用细胞原位杂交法观察不同培养时间 Ｔ 淋巴细胞 Ｂｃｌ２ ｍ Ｒ Ｎ Ａ 及 Ｂａｘｍ Ｒ Ｎ Ａ表达变化。结果　正常组及哮喘组 Ｔ 淋巴细胞抗 Ｃ Ｄ＋３ 单抗诱导后均出现凋亡典型形态改变,哮喘患者外周血 Ｔ细胞经抗 Ｃ Ｄ＋３ 单抗诱导２４ 小时后凋亡率为（９１９ ±２２５） ％ ,４８ 小时为（１５８９ ±２１８） ％ ,７２ 小时为（２１３７ ±３２４） ％ ;与正常组２４ 小时（１７９ ±２２２） ％ 、４８ 小时（２５２５ ±３５３） ％ 、７２ 小时（３５１４ ±２５３） ％ 比较,差异有显著性（ Ｐ 均＜ ００１） ,哮喘组７２ 小时方接近正常组２４ 小时水平。哮喘组 Ｔ 淋巴细胞凋亡抑制基因 Ｂｃｌ２ ｍ Ｒ Ｎ Ａ 的表达各时相均与正常组比较,差异有显著性（ Ｐ＜ ００１） 。而凋亡促进基因     

The autologous mixed lymphocyte reaction in primary biliary cirrhosis: analysis of activation and blastogenesis of autoreactive T lymphocytes

Blastogenesis of autoreactive T lymphocytes in the autologous mixed lymphocyte reaction has been shown to be decreased in patients with primary biliary cirrhosis, but the mechanisms underlying this abnormality are not known. To investigate further the abnormal autologous mixed lymphocyte reaction in primary biliary cirrhosis, we measured the activation of autoreactive helper/inducer and suppressor/cytotoxic T lymphocytes concurrently with blastogenesis in 35 patients with primary biliary cirrhosis and 18 healthy controls. Blastogenesis was diminished in autologous mixed lymphocyte reaction cultures from primary biliary cirrhosis patients compared to the control subjects (25,273 +/- 20,259 dpm vs. 36,004 +/- 14,951 dpm, p less than 0.02), but cultures from patients with primary biliary cirrhosis contained significantly increased percentages of activated helper/inducer and suppressor/cytotoxic T lymphocytes: 20.6 +/- 7.9 vs. 15.5 +/- 5.3%, p less than 0.008, and 9.8 +/- 7.8 vs. 5.4 +/- 3.0%, p less than 0.02, respectively. These findings were not related to the histologic stage of liver disease or to the serum bilirubin concentration and were not associated with abnormalities of lymphocyte subsets in fresh peripheral blood specimens. We conclude that the percentage of autoreactive T lymphocytes in autologous mixed lymphocyte reaction cultures from peripheral blood is increased in patients with primary biliary cirrhosis and diminished blastogenesis in autologous mixed lymphocyte reaction cultures is not due to the loss of autoreactive T lymphocytes from peripheral blood. Diminished blastogenesis reflects a diminished proliferative response of activated, autoreactive T lymphocytes in primary biliary cirrhosis. Possible mechanisms that may account for the paradoxical findings of decreased blastogenesis and increased activation of autoreactive T lymphocytes in primary biliary cirrhosis are discussed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dexamethasone inhibits CD4 T cell deletion mediated by macrophages from human immunodeficiency virus-infected persons.

Prednisolone slows the loss of CD4 T cells in individuals with human immunodeficiency virus (HIV) disease and inhibits antigen-induced apoptosis of recently HIV-infected CD4 cells in vitro. This study investigated whether dexamethasone inhibits the ability of macrophages to delete CD4 T cells via anti-CD4 antibody or immune-complexed HIV envelope protein gp120. Peripheral blood mononuclear cells from HIV-negative persons were incubated with CD4-reactive ch412 monoclonal antibody or with gp120/IgG immune complexes and resident macrophages, with and without dexamethasone. Dexamethasone inhibited CD4 cell deletion in a dose-dependent manner. The deletion of normal CD4 cells by macrophages from HIV-infected patients also was inhibited by dexamethasone. Furthermore, up-regulation of CD95 expression on T cells exposed to anti-CD4 and gp120/IgG, which predisposes T cells to CD95-mediated apoptosis, is inhibited by dexamethasone in a dose-dependent fashion. Dexamethasone inhibits the macrophage-mediated deletion of CD4 lymphocytes in HIV-infected persons.     

Effects of immunosuppressive therapy with prednisolone on B and T lymphocyte function in patients with chronic type B hepatitis

B and T lymphocyte function was studied in 10 patients with chronic type B hepatitis before, during and after a 28-day course of prednisolone therapy. Lymphocyte function was assessed by measuring the in vitro synthesis of immunoglobulin by peripheral blood mononuclear cells stimulated with pokeweed mitogen and by assaying lymphocyte proliferation in response to B and T cell mitogens. During high dose prednisolone therapy, there was a decrease in immunoglobulin synthesis by peripheral blood mononuclear cells and in lymphocyte proliferation to all mitogens. Studies using separated B and T cells showed that prednisolone treatment led to a decrease in both helper and suppressor T cell function but an enhancement of primary B cell function. When prednisolone was withdrawn, lymphocyte function rapidly returned to baseline levels. During prednisolone therapy, serum aminotransferase activities decreased by an average of 50%. In all patients, there was a subsequent rebound increase in serum aminotransferase activities 4 to 10 weeks after withdrawal of prednisolone. This was accompanied by a striking increase in suppressor T lymphocyte activity without significant changes in either helper T cell or B cell function. The close correlation between changes in helper and suppressor T lymphocyte function and serum aminotransferase activities during and after immunosuppressive therapy suggests that immunoregulatory T lymphocytes may play an important role in the pathogenesis of chronic type B hepatitis.     

T helper cell population and eosinophilia in nasal polyps.

OBJECTIVE: To analyze the immunological pattern of nasal polyposis in patients with and without allergy, the percentages of CD4+ cells expressing intracellular interferon-gamma and interleukin-4 (T helper [T(H)] type 1 and 2 cells) were measured by flow cytometry in samples from patients with nasal polyps. METHODS: Samples from 32 patients (16 atopic, 16 nonatopic) were studied. The fresh nasal polyp samples were prepared in single cell suspension for flow cytometry. Eosinophils were counted in hematoxylin-and-eosin-stained sections of all the samples. RESULTS: T(H)1 cells were predominant in all the nasal polyps, with no significant differences in the mean (+/-SD) percentages of T(H)1 cells between the 2 groups (46.28% +/- 14.95% vs 38.25% +/- 9.16%, P > .05). The mean percentage of T(H)2 cells in the polyps from the atopic patient group was significantly greater than in polyps from nonatopic group (7.34% +/- 2.54% vs 0.63% +/- 0.31%, respectively; P < .01); the eosinophil count was significantly higher in atopic patient polyp samples (54.5 +/- 15.76 eosinophils/HPF) than in nonatopic ones (14.38 +/- 5.6 eosinophils/HPF, P < .01). The mean percentage of T(H)1 cell correlated with eosinophil count in the polyp samples overall (r = 0.80, P < .01). CONCLUSIONS: T(H)1 cells were predominant in nasal polyp tissue. Polyps from atopic patients had more T(H)2 cells and eosinophils than nonatopic patients' polyps did. Eosinophil recruitment in nasal polyposis is probably associated with T(H)2 cell infiltration. Nonatopic and atopic patients' polyps have different immunological patterns.     

反义IL-5载体构建及其对IL-5 mRNA和蛋白表达的影响

目的构建含反义IL-5的重组腺相关病毒(rAAV)asIL-5,观察rAAV asIL-5对哮喘大鼠CD+4T淋巴细胞IL-5mRNA和蛋白表达的影响。方法用基因重组方法构建反义IL5rAAV真核表达载体质粒pasIL-5/rAAV,磷酸钙沉淀法将真核表达载体质粒pasIL-5/rAAV、包装质粒pXX2、辅助质粒pXX6共转染入病毒包装细胞293细胞中,合成rAAV asIL5,Southernblot测重组病毒的滴度;将rAAV asIL5转染经密度梯度法和免疫磁珠法分离的哮喘大鼠CD+4T淋巴细胞,用半定量RT PCR、ELISA分别检测转染后细胞内IL5mRNA及细胞培养上清液中IL-5蛋白的表达水平。结果(1)成功构建并鉴定了rAAV asIL-5,滴度为1.3×1011病毒颗粒/ml;(2)病毒转染孔的相对吸光度值为1.0515±0.1477,低于对照孔(1.4271±0.1655,P<0.01);(3)细胞培养上清液中IL-5的含量病毒转染孔为(12.0840±1.4769)ng/L,低于对照组[(15.3590±1.2685)ng/L,P<0.01]。结论rAAV asIL-5能够抑制哮喘大鼠CD+4T淋巴细胞IL5mRNA和蛋白表达,为研究哮喘的基因治疗提供了实验依据。