自身甲状腺球蛋白抗体阳性时甲状腺球蛋白测定方法的评估

应用单克隆固相放免法及双抗体法测定了自身免疫性甲状腺球蛋白抗体(TG-Ab)阳性病人血清中甲状腺球蛋白(TG)浓度。34例慢性淋巴细胞性甲状腺炎病人,TG-Ab阳性及TG-Ab阴性病人的TG值,分别为9.5±4.2ng/ml和10.9±3.7ng/ml(固相法);3.75±0.71ng/ml和4.25±1.1ng/ml(双抗体法)。50例Grave’s病人测定值分别为30.5±18.9ng/ml和41.5±20.5ng/ml(固相法);20.88±10.5ng/ml和27.5±12.5ng/ml(双抗体法)。38例甲状腺癌病人中,无转移灶者,两种方法测定值未见明显差异;8例有转移灶病人中,TG-Ab阳性者其TG值明显低于TG-Ab阴性者(p<0.01)。我们对10例慢性淋巴细胞性甲状腺炎TG-Ab阳性者进行了回收试验,固相法平均为81.6±14.0ng/ml,双抗体法平均为70.8±14.1ng/ml。我们认为,应用单克隆抗体固相法测定的TG值明显高于双抗体法,其回收率也高于双抗体法,但是在TG-Ab阳性病人血清中,TG测定值较TG-Ab阴性者为低,这可能由于在TG-Ab存在的情况下,体内TG代谢清除率增快而使TG浓度下降。因此,对TG-Ab阳性病人TG测定,仍有待于进一步探讨。     

BA-ELISA检测谷胱甘胱转移酶对肝癌的诊断意义

应用抗总的谷胱甘肽转移酶(EC 2.5.1.18 GSTs)的抗体,建立了生物素-亲和素-酶联免疫吸附法(BA-ELISA),并用以检测肿瘤病人血清中总GSTs浓度.本法非常敏感,其最低检出浓度可达0.05ng/ml,在0.05～6.0ng/ml的范围内,血清中总GSTs浓度和其相应之光密度值呈很好的直线关系.批内和批间的变异系数分别为8.2％和12.0％.肝癌、肝炎、消化道肿瘤、良性疾病患者血清总GSTs浓度分别为3.29±2.64,0.83±0.71,0.57±0.47和0.58±0.51ng/ml(mean±SD),而正常对照组仅为0.33±0.29ng/ml.肝癌病人血清总GSTs浓度显著高于其他疾病患者和正常人,总GSTs可望成为肝癌诊断的一个肿瘤标志。     

肾综合征出血热患者血浆和血清可溶性IL-2R水平变化的研究

本文应用双McAb ELISA夹心法检测了HFRS患者血清和血浆中SIL-2R水平。结果表明,急性期患者血浆和血清中sIL-2R水平(分别为199±81U/ml和214±122U/ml)显著地高于恢复期患者(分别为104±37U/ml和97±28U/ml)和正常人(分别为88±31U/ml和71±21U/ml)(P<0.01);在急性期中尤以少尿期升高最明显。表明HFRS患者机体免疫细胞处于高度活化的状态,患者血清和血浆中sIL-2R的检测对于判断病期和指导治疗均有一定的意义。     

Graves''病患者GH与IGF-1水平观察

目的:探讨Graves病(GD)患者对血清生长激素(GH)与胰岛素样生长因子-1(IGF-1)水平的影响.方法:采用放射免疫分析(RIA)检测42例GD患者、20例经治疗甲状腺功能恢复至正常水平GD患者与30例健康人血清GH与IGF-1水平.结果:GD组,血清IGF-1水平(170.8±44.4)ng/ml、GH水平(2.89±1.18)ng/ml明显升高,与对照组(IGF-1为90.5±30.5ng/ml、GH为1.58±1.20ng/m1)比较有显著性差异(p＜0.01、p＜0.05),且IGF-1与FT4正相关(r=0.58,p＜0.01).GD治疗缓解组,IGF-1水平(105.1±37.0)ng/ml、GH水平(1.71±1.36)ng/ml明显低于初诊未治疗组,差异有显著性(P＜0.01、P＜0.05).结论:GD患者血清IGF-1和GH水平明显升高,并且可能与甲状腺激素存在一定的关系.     

自然流产患者血清瘦素水平的改变

目的　探讨自然流产患者瘦素水平的变化及其临床意义。方法　用ELISA法测定 2 3名自然流产患者、2 1名正常妊娠者及 2 0名正常未孕者血清瘦素水平。结果　自然流产者血清瘦素水平平均为 875 3± 4 2 5 0ng/ml,正常妊娠者为1185 9± 4 0 3 2ng/ml,正常未孕者为 6 14 7± 15 0 2ng/ml,两两比较差异均有显著性。结论　自然流产患者血清瘦素水平明显降低 ,低瘦素水平可能是自然流产发生的又一原因。     

血清NSE,TNF-α,LSA联检对肺癌诊断的意义

目的 :探讨神经元特异性烯醇化酶 (NSE)、肿瘤坏死因子 -α(TNF -α)及脂质唾液酸 (LSA)在肺癌中的表达及其诊断价值。方法 :收集肺癌病人血清 78份 ,良性肺病患者血清 32份 ,正常对照血清 10 9份。采用酶联免疫吸附试验 (ELISA)和化学发光免疫分析法分别测定血清中的NSE、TNF -α及LSA。结果 :肺癌患者血清中的NSE、TNF -α、LSA分别为 19 78± 12 10ng/ml、135 6 4± 4 9 0 1pg/ml、1 0 6± 0 31mg/ml,明显高于良性肺病组 7 5 6± 3 4 1ng/ml、84 70± 2 4 89pg/ml、0 78± 0 18mg/ml及正常对照组 8 0 1± 2 81ng/ml、71 2 5± 13 5 0pg/ml、0 70± 0 13mg/ml(p <0 0 1)。对肺癌诊断的阳性率分别为 6 2 82 %、84 6 2 %、75 6 4 。联合检测阳性率达92 31%。结论 :血清NSE、TNF -α、LSA在肺癌病人中有较高的阳性率 ,联检阳性率更高 ,故NSE、TNF -α、LSA的联检是肺癌的非常有用的辅助诊断指标。 阳性标准为NSE >16 0ng/ml,TNF -α >10 0pg/ml,LSA >0 5 8mg/ml。     

溃疡性结肠炎患者PBMCs体外sIL-2R诱生水平的研究

目的探讨sIL-2R在溃疡性结肠炎（UC）中的表达水平。方法采用特异性ELISAsIL-2R检测试剂盒，对25例溃结患者PBMCs体外PHA诱导产生的sIL-2R水平进行测定。结果溃结患者PHA诱导48h的细胞培养上清液中sIL-2R的含量为319.63±164.70U/ml，明显低于正常对照组的451.15±246.93U/ml(P<0.05);而未经PHA诱导的sIL-2R含量，两组无明显差异（UC组263.78±115.05U/ml，对照组236.47±139.10U/ml，P＞0.05）。结论溃结患者体外PHA诱导的sIL-2R水平下降，提示其淋巴细胞的免疫活化功能障碍。     

新生儿缺氧缺血性脑病血浆ET-1与血清NSE动态变化及相关性的研究

目的:探讨新生儿缺氧缺血性脑病(Hypoxic-ischemicencephalopathy,HIE)血浆内皮素(Endothlin,ET)与血清神经元特异性烯醇化酶(Neuronspeificenolase,NSE)动态变化及相关性。方法:采用放射免疫分析动态测定32例HIE新生儿生后24h、3d、7d血浆ET-1和血清NSE水平,并以30例正常新生儿(生后24h)为正常对照组。结果:HIE新生儿血浆ET-1、血清NSE浓度在生后24h内分别为(88.17±19.75)pg/ml,(26.40±12.74)ng/ml显著高于对照组(分别为52.81±12.11pg/ml,14.35±3.11ng/ml);3d开始下降,血浆ET-1、血清NSE浓度分别为(73.77±22.86pg/ml,19.86±7.04ng/ml)与生后24h比较有显著性差异(P均<0.05);7d,HIE新生儿血浆ET-1(59.49±20.64)pg/ml,血清NSE(15.40±3.36)ng/ml与对照组比较无显著差异(P均>0.05)。血浆ET-1与血清NSE呈正相关(r=0.550,P<0.05)。结论:血浆ET-1动态变化在新生儿HIE的发病机理有着重要的作用,NSE释放与ET-1的动态变化相关。     

肠炎疾病IBD中结肠粘膜单个核细胞在体外释放可溶性IL-2受体增加

据报道Crohn's病(CD)和溃疡性结肠炎患者血清中可溶性IL-2受体(sIL-2R)浓度增高,在该文研究中,作者对正常及IBD结肠粘膜单个核细胞在体外自发释放sIL-2R的情况进行了观察.实验结果表明,CD患者结肠粘膜单个核细胞自发释放sIL-2R的量为204u/ml较正常对照组的释放量(124.5u/ml)明显增高(p<0.01),但溃疡性结肠炎患者sIL-2R释放量为135u/ml,与正常对照组相比无明显差异.此外,特异性疾病对照组的结肠     

肝病患者血清胰岛素样生长因子Ⅰ含量及与肝功能的关系

目的　研究肝病患者血清中胰岛素样生长因子I(IGF -I)的变化及其与肝功能的关系 .方法　各型肝炎患者 113例 ,对照组 30例 ,用放射免疫法检测血清IGF -I含量 ,同时检测有关肝功能指标 .结果　急性肝炎、慢性肝炎轻度、慢性肝炎中度患者血清IGF -I含量分别为 6 37.6± 178.3ng/ml、4 5 8.6± 2 76 .4ng/ml、6 0 1.7± 2 76 .8ng/ml,明显高于对照组 316 .1± 10 9.1ng/ml (p <0 .0 0 1) .慢性肝炎重度、肝硬化患者血清IGF -I含量为 2 2 4 .5± 16 8.2ng/ml和12 0 .5± 94 .4ng/ml明显低于对照组 (p<0 .0 5 ) .IGF -I含量与部分肝功能指标相关 (p <0 .0 5 ) .结论　检测IGF -I对估计慢性肝病预后有一定的价值 .     

他巴唑对Graves病患者血清可溶性白细胞介素—6受体的影响

５２例Ｇｒａｖｅｓ病（ＧＤ）患者随机分两组,２６例接受他巴唑（ＭＭＩ）、另２６例接受ＭＭＩ加心得安治疗。应用酶联免疫吸附分析（ＥＬＩＳＡ）,测定了未治疗期（第０周）,部分缓解期（第２周）和缓解期（第６周）患者及２０名健康对照者血清可溶性白细胞介素－６受体（ｓＩＬ－６Ｒ）水平。     

Increased and highly stable levels of functional soluble interleukin-6 receptor in sera of patients with monoclonal gammopathy

Soluble human interleukin-6 receptor (sIL-6R) was measured in the serum of 30 healthy individuals, 32 individuals with monoclonal gammopathy of undetermined significance (MGUS), 20 patients with early multiple myeloma (MM) and 54 patients with overt MM. The serum activity recognized by an immunoradiometric assay was determined to be sIL-6R, because of its binding capacity to IL-6 and its molecular mass of 55 kDa. All sera of healthy individuals contained sIL-6R (mean value: 89 ng/ml, range 17-300 ng/ml). Serum sIL-6R levels were increased by 51% in patients with MGUS (mean value: 135 ng/ml, p<0.005), by 44% in patients with early myeloma (mean value: 128 ng/ml, p<0.001) and by 116 % in patients with overt MM (mean value: 193 ng/ml, p<0.001). In patients with MM, a complete lack of correlation (p>0.7) was found between serum sIL-6R levels and other previously recognized prognostic factors in this disease, particularly serum IL-6 levels and those factors related to tumor cell mass. The independence of serum sIL-6R levels on tumor cell mass was directly demonstrated by studying four patients with MM treated with autologous bone marrow transplantation for periods of between 320 and 760 days. These levels were found to be remarkably stable and constant, independent of whether patients relapsed or achieved complete remission. Finally, physiological concentrations of sIL-6R were found to increase by tenfold the sensitivity of human myeloma cell lines to IL-6. These observations suggest a high control of the sIL-6R level in vivo, and, possibly, an important functional role of this circulating protein in patients with monoclonal gammopathies.     

Serum soluble interleukin-6 receptor in MRL/lpr mice is elevated with age and mediates the interleukin-6 signal

The characteristics of soluble interleukin-6 receptor (sIL-6R) in murine sera were examined. To investigate a relationship between serum sIL-6R level and autoimmune diseases, quantitative analysis of serum sIL-6R in MRL/lpr mice was performed by an enzyme-linked immunosorbent assay. The serum sIL-6R level in MRL/lpr mice of both sexes was below the detection limit (< 1.0 ng/ml) at 8 weeks of age, but it increased in accordance with age and reached 42 ± 9.3 ng/ml in female and 31 ± 13 ng/ml in male mice at 30 weeks of age. In MRL/+ mice, although an age-associated increase in serum sIL-6R level was observed, it was much less extensive than that in MRL/lpr mice. Elevated serum sIL-6R level at the age of 30 weeks was observed in female and male (NZB × NZW)F1 mice (32 ± 10 ng/ml and 17 ± 5.0 ng/ml, respectively), and male BXSB/Mpj Yaa mice (42 ± 18 ng/ml), suggesting that elevated serum sIL-6R in aged mice is one of the characteristics of autoimmune-prone mice. Quantitative analysis of serum IL-6 in MRL/lpr revealed that the serum sIL-6R level correlated well with the serum IL-6 level. We also showed that sIL-6R in the sera from MRL/lpr mice could mediate the IL-6 functions through the IL-6 signal-transducing receptor component gpl30, suggesting that elevated production of sIL-6R may partly contribute to development of autoimmune disease in MRL/lpr mice.     

The study of immunological markers in patients with "intrinsic" type atopic dermatitis]

Atopic dermatitis (AD) is a common inflammatory skin disease characterized by several clinical, immunological and biochemical alterations. Comparing the patients with the 'extrinsic' and 'intrinsic' types of AD, we investigated the role of immunological mechanisms in the pathogenesis of AD. To confirm it, we calculated serum markers of T lymphocyte activation: soluble interleukin-2 receptor (sIL-2R), interleukin-4 (IL-4), interleukin-10 (IL-10) and interferon-gamma (IFN-gamma). The soluble CD14 (sCD14) and tumor necrosis factor-alpha (TNF-alpha) in serum were measured as monocyte/macrophage activation markers. We examined 29 patients with the 'extrinsic' type AD (serum IgE > 10000 IU/ml: High-AD), 23 patients with the 'intrinsic' type AD (serum IgE < 37 IU/ml: Low-AD) and 11 healthy controls. Serum sIL-2R levels were increased in High-AD and Low-AD compared with the controls. They were also significantly increased in High-AD compared with Low-AD. Serum sCD14 levels were increased in High-AD compared with Low-AD and the controls. Severity index of AD were correlated with serum sIL-2R levels but not with sCD14 levels in sera. In conclusion, IgE may not relate with the pathogenesis of atopic dermatitis. Serum sIL-2R levels may be increased according to inflammatory skin lesions and it may be exaggerated with the immunological activation in the patients with the 'extrinsic' type AD.     

Clinical study on the serum carcinoembryonic antigen, immunosuppressive acidic protein and C-reactive protein levels in patients with adult T cell leukemia

Serum carcinoembryonic antigen (CEA), immunosuppressive acidic protein (IAP) and C-reactive protein (CRP) levels were measured in patients with adult T cell leukemia (ATL) in order to clarify their significance in this disease. Mean (+/- SD) serum CEA levels in 11 patients with acute ATL (3.1 +/- 1.3 ng/ml) and 7 patients with smoldering ATL (3.1 +/- 0.5 ng/ml) were significantly higher than in sera of 222 healthy controls (2.4 +/- 1.3 ng/ml). However, the levels in 7 patients with chronic ATL and healthy controls showed no differences. On the other hand, mean (+/- SD) serum IAP levels in patients with acute ATL (928 +/- 395 micrograms/ml), chronic ATL (487 +/- 125 micrograms/ml) and smoldering ATL (429 +/- 90 micrograms/ml) were significantly higher than in sera of healthy controls (359 +/- 103 micrograms/ml). However, the levels in patients with smoldering ATL and healthy controls showed no differences. Serum IAP levels in crisis in chronic and smoldering ATL were similar to those in patients with acute ATL. 85% of ATL patients with IAP levels above 500 micrograms/ml had CRP levels above 1+. Serum CEA, IAP and CRP levels were serially measured in a number of patients. Serum IAP and CRP levels reflected each patient's clinical course more than serum CEA levels. Overall the simultaneous measurements of serum CEA, IAP and CRP levels revealed a potential usefulness for determination of ATL subtype, and serum IAP and CRP levels may provide a way to evaluate the effectiveness of chemotherapy.     

Soluble interleukin-2 receptor and soluble CD8 antigen in active rheumatoid arthritis

Concentrations of soluble interleukin-2 receptor (sIL-2R) and of soluble CD8 antigen (sCD8) in sera and in supernatants of phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) derived from patients with active rheumatoid arthritis (RA) were studied. sIL-2R concentrations in sera derived from patients with RA (1484 +/- 382 U/ml) were significantly higher than in sera derived from healthy controls (380 +/- 110 U/ml; P less than 0.0005). In contrast, supernatants of PHA-stimulated PBMC derived from patients with RA contained similar amounts of sIL-2R (727 +/- 467 U/ml) as those derived from healthy control individuals (833 +/- 508 U/ml; P greater than 0.1). When investigated for the presence of sCD8 antigen, sera derived from patients with RA contained significantly lower amounts (30 +/- 28 U/ml) than sera derived from healthy controls (405 +/- 136 U/ml; P less than 0.0005). Similarly, PHA stimulation of PBMC derived from patients with RA resulted in a significantly lower production of sCD8 (35 +/- 46 U/ml) as compared to the one obtained by PHA stimulation of PBMC derived from healthy controls (177 +/- 59 U/ml; P less than 0.0005). This difference could not be explained by a lower proliferative response to PHA by PBMC derived from patients with RA (21,474 +/- 14,022 cpm) as compared to healthy controls (29,549 +/- 11,188 cpm; P greater than 0.05). Our data demonstrate that PBMC derived from patients with active RA differ from PBMC derived from healthy individuals concerning their ability to produce sIL-2R and sCD8.     

High-level production of alternatively spliced soluble interleukin-6 receptor in serum of patients with adult T-cell leukaemia/HTLV-I-associated myelopathy.

We have previously shown, using human T-cell lymphocytotrophic virus-I (HTLV-I)-infected cell lines, that soluble interleukin-6 receptor (sIL-6R) is generated through an alternative splicing mechanism. In this study, we examined human sera for the presence of alternatively spliced soluble IL-6R (AS-sIL-6R). We produced a monoclonal antibody (mAb) recognizing the unique sequence of AS-sIL-6R peptide, generated by an altered reading frame. We also made recombinant AS-sIL-6R protein in Spodoptera frugiperda-9 (Sf-9) cells carrying baculovirus, which encoded altered sIL-6R or conventional IL-6R cDNA. mAbs specifically recognized AS-sIL-6R, but not conventional IL-6R, as demonstrated by Western blot analyses, fluorescence-activated cell sorter, immunofluorescence analyses and enzyme-linked immunosorbent assay (ELISA). We adapted an ELISA system and used it for detection of altered sIL-6R in sera from 23 healthy persons, 12 patients with adult T-cell leukaemia (ATL) and 13 patients with HTLV-I-associated myelopathy (HAM). Serum levels of AS-sIL-6R were 6.4 or 6.1 times greater in ATL (28.7+/-20.4 ng/ml, P<0.0001) and in HAM patients (27.5+/-12.1 ng/ml, P<0.0001) than in healthy individuals (4.5+/-2.1 ng/ml). High levels of AS-sIL-6R were also observed in plasma from rheumatoid arthritis patients and in persons with elevated levels of alanine aminotransferase (ALT), antinuclear antibody (ANA), or alpha-fetoprotein (AFP). However, in human immunodeficiency virus-1 (HIV-1), hepatitis B virus (HBV) or hepatitis C virus (HCV)-infected individuals, AS-sIL-6R levels were not elevated. In this study, we confirmed that AS-sIL-6R is indeed present in human sera. These observations suggest that alternative splicing of IL-6R mRNA is of consequence in ATL, HAM and in some autoimmune diseases. The HTLV-I-infected T cells appeared to play an important role in AS-sIL-6R production.     

IL-6 and soluble IL-6 receptors (sIL-6R and sgp130) in human pleural effusions: massive IL-6 production independently of underlying diseases

IL-6, soluble IL-6 receptor (sIL-6R) and soluble gp130 (sgp130) levels were measured in sera and pleural effusions from 42 patients with metastatic carcinoma, non-Hodgkin's lymphoma, tuberculosis, cardiac failure and miscellaneous diseases. Pleural IL-6 levels measured by ELISA were very high in all patient groups (mean 34.8 ± 15.3 ng/ml) without significant difference according to diseases. IL-6 was shown to be biologically active in a proliferative assay. Serum IL-6 levels were low (0.049 ± 0.014 ng/ml) and did not correlate with pleural fluid levels. Pleural IL-6 levels correlated with the number of polymorphonuclear cells in pleural fluid (P< 0.03). Pleural sIL-6R levels (76 ± 8 ng/ml) were always lower than serum levels (196 ± 12 ng/ml; P< 0.0001) but correlated with them (P< 0.01). Pleural sIL-6R and albumin levels correlated (P< 0.01), suggesting a transudation of sIL-6R from the serum. Pleural sgp130 levels (10.9 ± 1.0 ng/ml) were lower than serum levels (24.6 ± 2.8 ng/ml; P< 0.002). After gel filtration of pleural fluid, the bulk of IL-6 (>90%) was recovered in a 15 000–30 000 fraction, corresponding to the expected mol. wt of free IL-6. These results suggest a production and a sequestration of IL-6 in the pleural cavity in all studied conditions.     

In vivo production of type 1 cytokines in healthy sickle cell disease patients

Interleukins (IL)-1, 2, 12, and interferon (IFN)-gamma, along with soluble IL-2 receptor (sIL-2R) were measured from sera obtained from healthy sickle cell disease (SCD) patients and comparable healthy control subjects. The cytokines were assessed by enzyme-linked immunosorbent assay (ELISA) in 60 SCD patients and 58 controls. No significant detectable levels of IL-1 or IL-12 were found in the sera of either group of patients. Significantly elevated levels of IFN-gamma were measured in 20 (33%) of 60 SCD patients and 21 (36%) of 58 controls. A large subset of 18 (41%) of 43 healthy controls and a smaller subset of 12 (21%) of 58 SCD demonstrated detectable levels of IL-2. The sIL-2R levels of the SCD group (4465 +/- 552 pg/mL) were significantly higher (P < .0001) than that of controls (3473 +/- 411 pg/mL). The results revealed comparable circulating levels of all type 1 cytokines in both healthy SCD and normal control subjects, with the exception of in vivo sIL-2R production. Elevated serum levels of both IL-6 and tumor necrosis factor (TNF)-alpha have been reported previously in a significant percentage of SCD steady-state subjects. These two cytokines are known to increase sIL-2R expression and may help explain the difference between the patient populations. Immune activation markers such as sIL-2R are produced by cells that mediate host responses to infection or inflammatory stimuli. The implication of higher levels of sIL-2R in SCD is not clear, but chronic parvovirus B19 infection, chronic polyclonal activation of B cells or defective regulation of antibodies are possible explanations for the elevated levels in SCD.     

Soluble interleukin-2 receptor in sera of patients with Graves' disease.

Activation of T lymphocytes has been found to be associated with an increase in soluble interleukin-2 receptor (sIL-2R) levels. The aim of this study was to investigate serum levels of sIL-2R in 20 untreated patients with Graves' disease and to relate these levels to disease activity and to TSH-receptor, anti-thyroglobulin, anti-microsomal and anti-eye muscle antibodies. sIL-2R levels were significantly increased in newly diagnosed Graves' patients compared with controls (667 +/- 270 vs 205 +/- 45 U/ml) (P less than 0.001). The sIL-2R levels were higher in patients with active infiltrative ophthalmology than in those without eye symptoms (810 +/- 313 vs 525 +/- 180 U/ml). All patients were treated with methimazole for at least 12 months. sIL-2R levels were normalized by methimazole treatment in the majority of patients without ophthalmopathy but not in those with ophthalmopathy. In five patients sIL-2R serum levels were studied after interruption of thyrostatic therapy. An increase was observed in three patients and hyperthyroidism subsequently relapsed in two of these. Furthermore, a correlation was found between soluble interleukin-2 receptor levels and TSH-receptor antibodies but not with other immune parameters examined. Serum sIL-2R represents a useful marker of immunological activity in Graves' disease.