普乐可复对难治性肾病综合征患者血清IL-2、sIL-2R的影响及其临床意义

目的:研究普乐可复(FK506)对难治性肾病综合征患者血清IL-2、sIL-2R的影响及其临床意义。方法:应用酶联免疫吸附法检测难治性肾病综合征患者经普乐可复治疗前后血清IL-2、sIL-2R水平变化,并监测患者24小时尿蛋白、血浆白蛋白、血脂的变化。结果:难治性肾病综合征患者经普乐可复治疗前血清IL-2、sIL-2R水平均显著高于正常对照组(P<0.05)。治疗后血清IL-2、sIL-2R水平较治疗前明显下降(P<0.05)。治疗前24小时尿蛋白水平显著高于正常对照组(P<0.01),血浆白蛋白显著低于正常对照组(P<0.01),血脂水平显著高于正常对照组(P<0.01);与治疗前比较,治疗后24小时尿蛋白水平显著下降(P<0.05),血浆白蛋白水平显著升高(P<0.01),血脂水平显著下降(P<0.05)。治疗后组与正常组比较,除IL-2外,余各项指标均无显著性差异(P>0.05)。结论:在难治性肾病综合征患者体内存在IL-2、sIL-2R的异常,普乐可复对其有明确的抑制作用,从而调节T细胞活性,有效降低24小时尿蛋白,提高血浆白蛋白含量,降血脂,缓解难治性肾病综合征的病情。     

一种检测可溶性白细胞介素2受体夹心法ELISA的建立

采用两种识别不同表位的小鼠抗人白细胞介素2受体(IL-2R)单克隆抗体(McAb),在国内首次建立了检测可溶性IL-2R(sIL-2R)的夹心法ELISA,并进行了初步应用。结果显示:本法特异性强、敏感性高、重复性好、操作简便,能检出正常人、某些患者血清中以及细胞培养上清中sIL-2R,可用于基础和临床免疫学研究。     

麻风病患者治愈后血清sIL-2R和TNFα的研究

目的研究麻风病病人治愈后免疫功能和细胞因子可溶性白细胞介素-2受体(sIL-2R)和肿瘤坏死因子α(TNFα)的潜在作用及调节异常,进一步探讨麻风病的免疫发病机理.方法对浙江省兰溪地区62例麻风病治愈者及其28例直系亲属的血清sIL-2R和TNFα进行检测,并以当地正常人作对照.结果各型麻风病治愈者血清TNFα和多菌型(MB)治愈者sIL-2R水平均明显降低(P＜0.001和P＜0.002),其中以瘤型麻风病(LL)治愈者TNFα和sIL-2R水平降低最显著(P＜0.001和P＜0.005),而少菌型(PB)治愈者sIL-2R与对照组无差异(P＞0.05).结论提示麻风病病人治愈后仍有细胞因子表达异常和单核巨噬细胞功能障碍,以及MB治愈者T细胞活化功能受抑制.这种血清TNFα和sIL-2R水平低下的持续存在,可能在麻风病病人对麻风杆菌的易感性及对麻风杆菌特异性细胞免疫缺陷的免疫发病中起重要作用.     

Perioperative serum levels of tumour-necrosis-factor alpha (TNF-alpha), IL-1 beta, IL-6, IL-10 and soluble IL-2 receptor in patients undergoing cardiac surgery with cardiopulmonary bypass without and with correction for haemodilution.

Cardiac surgery with cardiopulmonary bypass (CPB) leads to a systemic inflammatory response with secretion of cytokines. Alterations in the serum concentrations of cytokines have important prognostic significance. Reports on cytokine release during cardiac surgery with CPB have yielded conflicting results. Haemodilution occurs with the onset of CPB resulting in large fluid shifts during the perioperative course of cardiac procedures. In the present study we compare the perioperative course of serum concentrations of TNF-alpha, IL-1beta, IL-6, IL-10 and sIL-2R with and without correction for haemodilution in patients undergoing coronary artery bypass grafting (CABG) surgery. Twenty male patients undergoing elective CABG surgery with CPB and general anaesthesia using a balanced technique with sufentanil, isoflurane and midazolam were enrolled into the study. Serum levels of TNF-alpha, IL-1beta, IL-6, IL-10 and sIL-2R were measured using commercially available ELISA kits. Simultaneous haematocrit values were obtained at all sample times. Statistical analysis was performed by non-parametric analysis of variance and t-tests for data corrected for haemodilution and data that were not corrected for haemodilution. Adjusted significance level was P < 0.01. Intra-operatively, up to the second post-operative day PCV values were significantly decreased compared with preoperative values. Cytokine measurements not corrected for haemodilution were significantly lower than the corrected values. The perioperative haemodilution and decrease in PCV may lead to an underestimation of the cytokine secretion in post-operative patients. We conclude that cytokine measurements were significantly influenced by the perioperative haemodilution and the subsequent decrease in PCV and may in part account for the varying results reported in the literature regarding cytokine release in patients undergoing CABG surgery.     

High levels of plasma IL-10 and expression of IL-10 by keratinocytes during visceral leishmaniasis predict subsequent development of post-kala-azar dermal leishmaniasis

Some patients develop post-kala-azar dermal leishmaniasis (PKDL) after they have been treated for the systemic infection kala-azar (visceral leishmaniasis). It has been an enigma why the parasites cause skin symptoms after the patients have been successfully treated for the systemic disease. We report here that PKDL development can be predicted before treatment of visceral leishmaniasis, and that IL-10 is involved in the pathogenesis. Before treatment of visceral leishmaniasis, Leishmania parasites were present in skin which appeared normal on all patients. However, IL-10 was detected in the keratinocytes and/or sweat glands of all patients who later developed PKDL (group 1) and not in any of the patients who did not develop PKDL (group 2). Furthermore, the levels of IL-10 in plasma as well as in peripheral blood mononuclear cell culture supernatants were higher in group 1 than in group 2.     

Enhancement of IL-1, IL-2 production and IL-2 receptor generation in patients with acute rheumatic fever and active rheumatic heart disease; a prospective study.

In a prospective study, patients with quiescent rheumatic heart disease (CRHD), streptococcal pharyngitis (SP) and healthy normal subjects produced comparable amounts of IL-1 and IL-2, but acute rheumatic fever (ARF) patients produced significantly elevated amounts of IL-1 and IL-2 at all intervals up to 48 weeks. In active rheumatic heart disease (ARHD), IL-1 activity returned to within normal range at 48 weeks, but IL-2 activity remained persistently elevated compared with CRHD, SP and healthy age- and sex-matched volunteers. CD4+ T lymphocytes were significantly increased in the peripheral blood of ARF and ARHD patients. The amount of IL-2 produced by ARF and ARHD patients correlated with the percentage of helper T lymphocytes (CD4+ cells) but not with the percentage of suppressor/cytotoxic T lymphocytes (CD8+ cells). Moreover, pre- and post-phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cell (PBMC) cultures from ARF and ARHD patients contained higher proportions of IL-2R+ (CD25+) cells than those from patients with SP, CRHD and normal individuals, which persisted up to 48 weeks. The percentage of CD25+ cells in both types of PBMC cultures directly correlated with the percentage of CD4+ cells and not with CD8+ cells in active rheumatic patients only. These findings indicate that the immune response in ARF and ARHD patients is skewed to produce activated helper T cells that release IL-2 which drives the accumulation of more T helper cells. The result is an undamped helper T cell response in the peripheral blood of these patients.     

The correlation between interleukin 2 and soluble interleukin 2 receptors to oestradiol, progesterone and testosterone levels in periovulatory follicles of in-vitro fertilization patients.

The present study was performed to evaluate the correlation between follicular fluid levels of interleukin 2 (IL-2) and IL-2 soluble receptor (sIL-2R), oestradiol, progesterone and testosterone levels, oocyte fertilization, embryo quality and pregnancy rates. Twenty-eight patients with a pure tubal factor and undergoing in-vitro fertilization and embryo transfer were randomly chosen and treated with gonadotrophin releasing hormone agonist (GnRHa) in the midluteal phase (long protocol) coupled with follicular phase administration of human menopausal gonadotrophin. Transvaginal follicular aspiration was performed 36 h after human chorionic gonadotrophin administration, followed 48 h later by embryo transfer. One hundred and twenty-three follicular fluids were sampled. The mean follicular fluid levels (+/- SD) were 2.30 +/- 0.80 fmol for IL-2, 458.2 +/- 236.0 units/ml for sIL-2R, 28.5 +/- 58.1 ng/ml for oestradiol, 2360.5 +/- 2846 ng/ml for progesterone and 7.22 +/- 7.08 ng/ml for testosterone. There was a significant (P less than 0.01) correlation between IL-2 and testosterone levels. No correlation was found between the lymphokines and serum oestradiol, follicular fluid progesterone, oocyte fertilization, embryo quality and pregnancy. It may be concluded that significant concentrations of IL-2 and sIL-2R exist in follicular fluid. Wide variations in follicular IL-2 and sIL-2R concentrations of different follicles were found in the same patients.     

Evaluation of serum levels of soluble CD4, CD8 and beta 2-microglobulin in visceral human leishmaniasis.

The levels of soluble CD4 (sCD4), sCD8 and beta 2-microglobulin (beta 2-M) were measured in sera from patients with visceral leishmaniasis during the course of infection. Levels of sCD4, sCD8 and beta 2-M were raised significantly above levels in normal sera and returned to the normal range after recovery. The decrease in the levels of sCD8 was related to a reduction of anaemia, leukopenia and thrombocytopenia. In contrast, sCD4 levels fluctuated during the period of infection. beta 2-M returned within normal range more rapidly than sCD8 secretion. Our results suggest that T cells are activated during infection, and that it is also possible that the raised levels of these soluble molecules play a role in the impairment of protective immunity.     

IL-2 regulation of soluble IL-2 receptor levels following thermal injury.

In the immunosuppressed burn patient serum levels of both IL-2 and a soluble form of IL-2 receptor alpha (sIL-2R alpha) are significantly elevated. Strikingly, the production of these markers by the in vitro activated patients' cells is decreased. This study examines the role of IL-2 in the decreased production of the sIL-2R alpha in vitro in patients with major burns (n = 18, 30 to greater than 70% total body surface area). Peripheral blood mononuclear cell (PBMC) cultures from patients with highly elevated serum sIL-2R alpha, and from healthy controls (n = 12) were activated with concanavalin A (Con A) at initiation. In patients' cultures mitogen-induced increments of sIL-2R alpha levels were significantly lower. There was a significant negative correlation (r = 0.64, P less than 0.001) between a high serum sIL-2R alpha level and a decreased lectin-induced sIL-2R alpha release in vitro. Low levels of sIL-2R alpha in patients' samples were not normalized by increasing the number of T lymphocytes. Also exogenous rIL-1 was without effect, whereas rIL-3 increased sIL-2R alpha release in some cultures. However, sIL-2R alpha levels were significantly increased in patients' cultures by (i) addition of exogenous IL-2; (ii) removal of adherent cells; (iii) addition of cyclooxygenase inhibitor, indomethacin; (iv) bypassing cell surface activation by the combination of the calcium ionophore A23187 and the phorbol ester 12-o-tetradecanoyl acetate. The cyclic AMP-elevating drug, forskolin, abrogated the ability of exogenous IL-2 to increase sIL-2R alpha production. Thus, in the burn patient, the reduced in vitro sIL-2R alpha release appears to relate to abnormalities in IL-2 production and action mediated through its functional surface receptor. Elevated levels of sIL-2R alpha in vivo may, therefore, reflect systemic activation of T lymphocytes in response to biologically active IL-2.     

不同患者手术前后血中IL-2/sIL-2R系统的动态变化

本文研究28例胆囊炎胆石症、结肠癌和直肠癌患者手术前后血中IL-2/sIL-2R系统的动态变化。分别于手术前或当晨,术后1、2～3、5～7d抽血,测定血中IL-2、sIL-2R量,同时作CRP试验。结果表明,施行中等手术胆囊摘除总胆管切开引流术和半结肠切除术对IL-2/sIL-2R系统的影响与患者术前的水平有关。原先异常程度或轻或重,术后影响亦或轻或重,但均未见统计学差异。3例直肠癌患者施行miles氏手术,术后出现明显IL-2量下降,sIL-2R量增高,持续时间长。因此,初步说明创伤大的手术在一定程度上影响机体的IL-2/sIL-2R系统。并可从中发现规律,采取措施,具有一定的临床价值。     

抽动秽语综合征患者血清自身抗体与可溶性白介素-6受体表达增高

目的探讨抽动秽语综合征(TS)病程中自身免疫损伤相关机制。方法用ELISA法检测TS患者血清中抗脑抗体(ABAb)、抗核抗体(ANAb)、可溶性白介素-6受体(sIL-6R)及可溶性gp130(sgp130)的含量,用胶乳增强免疫比浊法检测抗链球菌溶血素O抗体(ASO)水平。结果TS组血清sIL-6R和sgp130含量显著高于对照组[(44.1±15.8)ng/mLvs(30.3±9.0)ng/mL和(69.0±24.6)ng/mLvs(47.3±14.1)ng/mL,P<0.01];血清ABAb和ANAb检出率显著高于对照组(66%vs4%和53%vs25%,P<0.01),并且ASO增高的比例高于对照组(P<0.01);TS组ABAb水平与sgp130呈负相关(r=0.375,P<0.05)。结论TS患者依赖IL-6信号传导的生理功能上调和反馈抑制的网络机制启动;自身免疫相关的损伤机制均可能参与了疾病发生发展的病理生理过程。     

可溶性白细胞介素2受体与白血病相关性的初步研究

检测34例白血病患者的血清可溶性白细胞介素2受体(sIL—2R)水平,並分析了sIL—2R与病情变化的相关性。结果显示:1.各组患者的sIL—2R水平明显高于正常(p<0.01);2.sIL-2R水平>2000u/ml的6名患者中有5名在采血后1个月内死亡,余1例亦在1年内死亡;3.患者血清sIL—2R水平与末梢血白细胞总数及单个核细胞数无明显关系。分析表明:sIL—2R测定适用于对急性淋巴细胞性、单核细胞性白血病及淋巴瘤合并白血病的病情监测及预后判断。     

新生儿脐血T细胞IL－2R表达和血清sIL－2R水平的研究

对新生儿脐血Ｔ细胞ＩＬ－２Ｒ表达率和血清ｓＩＬ－２Ｒ水平进行检测，结果显示：①早产儿脐血Ｔ细胞ＩＬ－２Ｒ自然表达率明显低于足月新生儿和正常儿童组，而后两者间无差异；②经ＰＨＡ活化后足月新生儿和早产儿Ｔ细胞ＩＬ－２Ｒ表达率皆显著低于正常儿童组，早产儿组也明显低于足月新生儿组；③足月新生儿和早产儿脐血血清ｓＩＬ－２Ｒ水平均明显高于正常儿童组，但前两者间无明显差异；④新生儿组（包括足月和早产儿）脐血Ｔ细胞ＩＬ－２Ｒ自然表达率与血清ｓＩＬ－２Ｒ水平呈直线负相关，而正常儿童组二者间呈直线正相关。     

过敏性紫癜患儿血清sIL-2R、IL-6、IL-8 及凝血功能的变化

目的　探讨过敏性紫癜 (HSP)患儿血清可溶性白介素 2受体 (sIL - 2R)、白介素 6 (IL - 6 )、白介素 8(IL - 8)及部分凝血活酶时间 (APTT)、凝血酶时间 (TT)、D -D二聚体在发病过程中的变化。方法　应用化学发光法检测sIL - 2R、IL -6、IL - 8水平 ,用SymexCA - 15 0 0全自动血凝分析仪测定APTT、TT及D -D二聚体。结果　HSP患儿急性期血清sIL - 2R、IL- 6、IL - 8水平明显高于正常 (P <0 0 5 ) ,3个月后恢复至正常对照组水平 (P >0 0 5 )。早期血液呈高凝状态 ,APTT、TT缩短 ,伴继发性纤溶 ,D -D二聚体增高 ,且混合型增高更显著。结论　动态检测HSP患儿血清sIL - 2R、IL - 6、IL - 8及APTT、TT、D -D二聚体的变化 ,为临床判断病情 ,估计预后及抗凝治疗提供理论依据。     

Follow up of soluble IL-2 receptor level in metastatic malignant melanoma patients treated by chemoimmunotherapy.

Immunological parameters following chemoimmunotherapy combination were studied in 31 patients with metastatic malignant melanoma. They received Cisplatin (100 mg/m2) on day 1 and 28, recombinant IL-2 (rIL-2; Eurocetus) in continuous infusion from day 3 to 6, 17 to 21, 31 to 34 and 45 to 49. Interferon-alpha (IFN-alpha; Roche) was given subcutaneously three times weekly. No significant change in CD4/CD8 ratio at onset or during treatment was observed between responder (n = 19) and non-responder (n = 12) patients. Regarding the IL-2 receptor (IL-2R) study, the percentage of cells expressing Tac (p55) receptor did not change either for healthy volunteers (n = 20) and patients before any therapy, or between responder and non-responder patients. Concerning serum soluble IL-2R shedding before therapy, we observed a significant increase (P = 0.001) in patients (79 +/- 40 pM) compared with healthy donors (30 +/- 15 pM), but no significant variation was seen between responder and non-responder patients. In contrast, during the treatment, the soluble IL-2R level increased in both groups but, interestingly, a significant difference was found between responder and non-responder patients from day 7 (P < 0.05) to day 21 (P < or = 0.01), suggesting that the cells from non-responder may be slower in becoming stimulated. This finding is the most striking point of our study and suggests that sIL-2R might be an early predictive factor of the clinical response as obtained by logistic regression (P = 0.0063). Therefore patients with a serum soluble IL-2R level greater than 250 pM at day 21 have a 12-fold more chance of undergoing a clinical response.     

A combination therapy using IL-12 and soluble IL-4 receptor on herpes simplex virus Type 1 infection in a human-SCID chimera model of thermal injury

Herpes simplex virus type 1 (HSV-1) is a severe pathogen in thermally injured patients. Type 1 T cells are essential for the host's anti-HSV protective immunity. Type 2 cytokines, commonly detected in thermally injured patients, have been described as inhibitors for the type 1 T cell generation. Therefore, the antiviral effects of combination therapy with a type 1 T cell inducer [interleukin (IL)-12] and a type 2 T cell inhibitor [soluble IL-4 receptor (sIL-4R)] were investigated in severe combined immunodeficiency (SCID) mice inoculated with peripheral blood lymphocytes (PBL) of thermally injured patients. Patient PBL-SCID chimeras (SCID mice inoculated with patient PBL) were susceptible to infection with 1 x 10(3) PFU/kg of HSV-1 (0% survival), while healthy PBL-SCID chimeras (SCID mice inoculated with PBL from healthy donors) were resistant (92% survival). When patient PBL-SCID chimeras exposed to HSV-1 were treated with saline, human recombinant (r) IL-12 or human sIL-4R, 0, 0, or 12.5% of them survived, respectively. However, 75% of these chimeras survived when they were treated with rIL-12 and sIL-4R in combination. These results indicate that HSV-1 infection in patient PBL-SCID chimeras was therapeutically controlled by the inducer of type 1 T cell responses and the inhibitor of type 2 T cell responses in combination.     

乙型肝炎患者可溶性白细胞介素2受体水平变化

本研究应用酶联免疫吸附试验(ELISA)检测了103例乙型肝炎各型患者及26倒乙肝病毒(HBV)携带者血清中可溶性白细胞介素2受体(sIL-2R)水平,并结合临床资料进行分析。结果发现:各型患者及HBV 携带者sIL-2R 水平均明显高于正常对照(P<0.01～0.001),其升高程度与病情轻重呈正比,在去除危重患者酶—胆分离时谷丙转氨酶(ALT)过低的影响后,sIL-2R 水平与ALT 有良好的正相关关系(r=0.38.p<0.05);作者认为sIL-2R 能够较好地反映疾病的活动状况,动态观察血清sIL-2R 水平的变化,对判断和评价HBV 感染后的病情演变及免疫失衡状态具有重要意义。     

Serum soluble markers in the evaluation of treatment in human visceral leishmaniasis.

Visceral leishmaniasis (VL) has a fatal course if not properly treated. Recovery from VL is linked to cellular immune response. Unresponsiveness to antimonial therapy reinforces the importance of determining parameters for treatment assessment. We analysed the pre- and post-treatment serum levels of soluble CD4 (sCD4), sCD8, sIL-2R, soluble intercellular adhesion molecule-1 (sICAM-1) and neopterin in groups of VL patients either responsive or not to standard antimonial therapy. Pretreatment serum levels of all markers except for sICAM-1 were significantly higher in VL patients than in healthy subjects from the same area (P < 0.05). sICAM-1 levels were similar in healthy controls and in VL patients refractory to antimonial therapy (P = 0.25), but significantly higher in patients responsive to treatment (P = 0.02). The comparison of pre- and post-treatment concentrations showed that all markers, except sCD4 and sICAM-1, presented a significant fall (P < 0.05) in patients responsive to antimonial therapy. However, only neopterin presented with levels compatible with those of healthy subjects at the end of treatment (P = 0.30). In refractory patients sICAM-1 presented with post-treatment levels significantly higher than the pretreatment determinations (P = 0.03), while sCD4 experienced a significant drop (P = 0.01). All markers displayed clearly distinct behaviour according to the patient's response to therapy. This makes all soluble molecules studied suitable for use as indicators of antimonial therapy response. Additionally the comparison of pretreatment levels of the markers between responders and refractory patients to antimonial therapy showed that serum concentrations of sIL-2R and sICAM-1 significantly differed among these two groups (P = 0.02 in each case), suggesting that they may be used in future as predictors of antimonial therapy response.     

慢性肾炎患者血清sIL－2R和IL－8的变化及意义

研究用ＥＬＩＳＡ双抗体夹心法对慢性肾炎肾功能不全各期患者血清ｓＩＬ－２Ｒ和ＩＬ－８水平进行了测定。结果显示：①肾功能不全代偿期ｓＩＬ－２Ｒ水平显著高于健康人（Ｐ＜０．０１），提示此期存在Ｔ细胞异常活化。②氮质血症期ｓＩＬ－２Ｒ水平显著高于健康人和代偿期（Ｐ＜０．０１），与Ｓｃｒ呈显著正相关（ｒ＝０．６５，Ｐ＜０．０００５），提示血清ｓＩＬ－２Ｒ水平可作为肾小球肾炎恶化的重要指标。③尿毒症期ＩＬ－８水平显著低于健康人（Ｐ＜０．０１），ｓＩＬ－２Ｒ水平显著低于氮质血症期（Ｐ＜０．０１），提示此期存在细胞免疫缺陷，其中单核细胞缺陷可能是Ｔ细胞功能低下的重要原因；ＩＬ－８水平降低可能是此期中性粒细胞吞噬功能减弱的原因之一。总之，本研究有助于进一步阐明慢性肾炎的发病机理。     

Serum levels of soluble IL-2 receptor, IL-4 and IgE-binding factors in childhood allergic diseases.

The serum levels of soluble IL-2 receptor (sIL-2R), IL-4 and IgE-binding factors were examined in children with allergic diseases, and compared with those in non-allergic controls of the same age and sex. The results showed age-related decreases in the serum levels of sIL-2R and IgE-binding factors, but not in that of IL-4 in both allergic and non-allergic individuals. Significant elevation of sIL-2R was observed in sera from children with atopic eczema or history of an anaphylactic reaction to food, as compared with that in non-allergic controls. The serum concentration of IL-4 was elevated in all allergic groups, including cases of atopic eczema, bronchial asthma and anaphylaxis to food, compared with non-allergic controls, and was correlated significantly with the serum level of IgE (r = 0.59). The IgE-binding factor levels in sera from patients aged 6-10 years with bronchial asthma, or patients aged 1-5 years with a history of food anaphylaxis were elevated as compared with those in non-allergic controls of same age. There was no significant correlation between the serum levels of IgE-binding factors and IgE. Since sIL-2R is released by activated T cells, the present study is in favour of T cell activation causing allergic skin disorders. The serum levels of IL-4 as well as IgE did not differ among allergic patients of different clinical categories. The role of IgE in atopic eczema and other allergic diseases is not clearly established; however, it seems likely that IL-4 is deeply involved in the increased production of IgE seen in allergic individuals. The possible involvement of IgE-binding factors in the age-related changes of clinical manifestations in childhood allergic diseases was also discussed.