IL-1β、NO、TNF-α、瘦素对妊娠期高血压疾病的临床研究

目的探讨妊娠期高血压疾病患者白介素-1β（IL-1β）、一氧化氮（NO）、肿瘤坏死因子（TNF-α）、瘦素（Leptin）的相关性。方法选取2010年6月～2012年12月在我院住院妊娠期高血压疾病（HDCP）患者70例,根据病情分为妊娠期高血压组（GH）30例,轻度子痫前期组（MP）20例,重度子痫前期组（SP）20例,同期妊娠末期健康孕妇30例为对照组。监测IL-1β、NO、TNF-α、瘦素的测定。结果妊娠期高血压疾病组NO浓度水平显著低于对照组,TNF-α、Leptin血清水平明显高于对照组,妊娠期高血压疾病患者胎盘组织中白介素-1β（IL-1β）mRNA水平显著高于正常孕妇。差异有统计学意义。结论 IL-1β、NO、TNF-α、Leptin的测定对妊娠高血压疾病的筛查诊断治疗有价值,可作为临床应用。     

Th17细胞在子痫前期患者外周血表达增加

观察正常妊娠妇女和子痫前期疾病患者外周血CD4+T细胞中Th17细胞和特异性转录因子维A酸相关孤独受体(retinoid-related orphan nuclear receptor,RORγt)的表达和意义。实验组为子痫前期疾病患者25例,正常妊娠妇女20例,未孕妇女20例。采集研究对象外周血,酶联免疫吸附试验(ELISA)检测Th17细胞相关细胞因子IL-17A,IL-6,TNF-α的表达,Ficoll法分离外周血单个核细胞(PBMC),免疫磁珠分选CD4+T淋巴细胞,逆转录-聚合酶联反应(RT-RCR)半定量检测CD4+T细胞中Th17细胞特异性转录因子RORγt的表达,流式细胞术检测CD4+IL-17+T细胞比例。子痫前期患者外周血清中IL-6、IL-17A的含量分别为(31.72±13.34)ng/L、(2.61±1.64)ng/L,高于正常妊娠组水平,差异有显著统计学意义(P<0.01),TNF-α在子二组间的表达分别为(18.00±8.64)ng/L和(11.69±3.68)ng/L,差异有统计学意义(P<0.05);RORγt mRNA在子痫前期组的表达高于正常妊娠组,净光密度值差异有显著统计学差异(P<0.01),CD4+IL-17+T细胞在子痫前期组的表达为(1.83±0.42)%,高于正常妊娠组(0.87±0.26)%,差异有显著统计学意义(P<0.01)。子痫前期患者外周血CD4+T细胞中RORγt mRNA、Th17细胞以及Th17细胞相关细胞因子表达异常,可能在疾病的发病机理中起到重要作用。     

IL-2、IL-10、TNF-α在子痫前期不同发病类型中的水平变化及意义

目的探讨IL-2、IL-10、TNF-α在子痫前期不同发病类型患者血清中的水平变化。方法用ELASA检测30例早发型重度子痫前期、30例晚发型重度子痫前期和30例正常妊娠孕妇血清中IL-2、IL-10、TNF-α的水平。结果早发型重度子痫组和晚发型子痫前期组患者体内IL-2、TNF-α水平与对照组相比明显升高,差异有显著统计学差异(P0.05),早发型重度子痫组和晚发型子痫前期组中IL-2、IFN-α水平相比也有显著统计学差异(P0.05);早发型重度子痫组和晚发型子痫前期组患者体内IL-10水平与对照组相比明显降低差异有统计学意义(P0.05),而早发型重度子痫组和晚发型子痫前期组患者体内IL-10水平相比差异无统计学意义(P0.05)。结论子痫前期患者血清中的IL-2、TNF-α的水平升高及IL-10水平降低与该病的发生发展密切相关。     

IL-1β TNF-α, and IL-2 in Peritoneal Fluid and Macrophage-Conditioned Media of Women With Endometriosis

PROBLEM : The presence of various cytokines in human peritoneal fluid has been incompletely evaluated. Changes in cytokine levels may be related to the development of endometriosis, infertility, and activation of peritoneal macrophages. This study assesses levels of IL-1β, IL-2, and TNF-α in peritoneal fluid and macrophage conditioned media of women with endometriosis. METHOD : Peritoneal fluid was collected from 51 women at the time of diagnostic or operative laparoscopy for benign gynecologic disease. Peritoneal macrophages were isolated, cultured for 24 h, and the culture media collected. IL-1β, IL-2, and TNF-α levels were determined by commercial ELISA kits. RESULTS : The mean concentration of IL-1β and TNF-α was significantly higher in macrophage conditioned media of patients with endometriosis (P < 0.02). However, there were no significant changes in peritoneal fluid cytokine levels. Peritoneal macrophage concentrations were also higher in patients with endometriosis. CONCLUSION : This study supports the concept that endometriosis is associated with activation of peritoneal macrophages, and a higher concentration of these cells. This activation is reflected by the increased levels of cytokines found in macrophage conditioned media. The absence of significant changes in peritoneal fluid cytokine levels would seem to indicate that the above derangements are not responsible for the development or progression of endometriosis.     

Correlation of P-selectin and lipoprotein(a), and other lipid parameters in preeclampsia

Preeclampsia is a pregnancy specific disorder and is thought to be associated with generalized endothelial dysfunction. P-selectin, an adhesion molecule, mediates the interaction of monocytes, platelets, and endothelial cells. Increased P-selectin levels and altered lipid and lipoprotein metabolism were reported in preeclampsia and during pregnancy. In order to investigate the relationship between serum P-selectin and lipoprotein(a), and other lipid parameters, 28 preeclampsia [13 severe (group I) and 15 mild preeclampsia (group II), 15 healthy pregnant (group III) and 20 non-pregnant (group IV)] women were investigated. Serum P-selectin, lipoprotein(a), total cholesterol, triglyceride, and high density lipoprotein cholesterol were measured and low-density lipoprotein cholesterol was derived. Serum P-selectin concentrations were consistently and significantly higher in the severe preeclampsia group than in the mild preeclampsia, healthy pregnancy, and non-pregnant control groups (P<0.0001, for all). The mild preeclampsia group also had increased serum P-selectin concentrations compared with the healthy pregnancy group and non-pregnant controls (P<0.05 and P<0.0001, respectively). Serum P-selectin and lipoprotein(a) levels revealed a significant and linear increase with the severity of preeclampsia. There were also significant (in groups I and II) and borderline (in groups III and IV) correlations between P-selectin and total cholesterol. The present study suggests that P-selectin may be an additional risk marker for preeclampsia, and may be useful in distinguishing women with mild and severe preeclampsia and normal pregnancy. Received: 9 November 2001 / Accepted: 6 February 2002     

Effect of Human Chorionic Gonadotropin on Cytokine Production from Human Endometrial Cells in Vitro

PROBLEM: To examine whether human chorionic gonadotropin (hCG) is involved in the regulation of interleukin (IL)-6, tumor necrosis factor (TNF)-α, and leukemia inhibitory factor (LIF) secretion from cultured human endometrial cells. METHOD OF STUDY : A mixed population of endometrial cells from six in vitro fertilization/embryo transfer patients was cultured and incubated with various doses of hCG (0, 1, 10, 50, 100, and 500 IU/ml) for 24 hr. IL-6, TNF-α, and LIF levels in the culture medium were measured with enzyme-linked immunosorbent assay. RESULTS: IL-6 and TNF-α levels were stimulated by hCG in a dose-dependent manner. Stimulation of IL-6 and TNF-α levels by 500 IU/ml of hCG increased their production by 3.7- and 2.8-fold, respectively (P < 0.05). Stimulation of IL-6 by 100 IU/ml of hCG was also significant (P < 0.05). However, there was no significant effect of hCG on LIF secretion by endometrial cells (P = 0.31). CONCLUSIONS : hCG is involved in the regulation of endometrial cytokine production from human endometrial cells in vitro. This finding supports the recently emerging notion that hCG could have important local roles within the uterus besides its well-known luteotrophic role on the corpus luteum for maintenance of pregnancy.     

Circulating Tumor Necrosis Factor (TNF)-α in Normally Cycling Women and Patients with Premature Ovarian Failure and Polycystic Ovaries

PROBLEM : The purpose of this study was to investigate the levels of tumor necrosis factor (TNF)-α in sera of infertile patients having premature ovarian failure (POF) and polycystic ovaries (PCO), and to compare the levels with those in age-matched normally cycling women. METHOD : Levels of TNF-α, luteinizing hormone (LH), follicle-stimulating hormone (FSH), and estradiol (E2) were measured in the sera of infertile and normal women by the enzyme linked immunosorbent assay (ELISA), fluorescent immunoassay and radioimmunoassay (RIA), respectively. Multiple groups comparisons were performed using Dunnet's test. Post-hoc analysis was performed using a Tukey-Kramer test. P < 0.05 was considered statistically significant. RESULTS : TNF-α levels were significantly (P = 0.003 to < .0001) lower in the sera of patients with POF compared to sera of controls and patients with PCO. CONCLUSIONS : TNF-α may have a role in ovarian function. Derangement in follicular development, luteal function and atresia results in a change of circulating TNF-αa levels.     

The Effect of SHJKS on Cytokines Production and NF-κB Activation in the Peripheral Blood Mononuclear Cells of Patients with Cerebral Infarction

The Korean genuine medicine “Seonghyangjeongkisan” (SHJKS) has long been used for various cerebrovascular diseases. However, very little scientific investigation has been carried out. Cytokines involved in the regulation of inflammatory reactions and immune responses may play a role in the pathogenesis of cerebral infarction (CI). The aim of the present study is to elucidate how SHJKS modulates the inflammatory reaction in lipopolysaccaride (LPS) plus phytohaemagglutinin (PHA)-stimulated peripheral mononuclear cells (PBMCs) from CI patients. The amount of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-8 in PBMC culture supernatant was significantly increased in the LPS plus PHA treated cells compared to unstimulated cells. SHJKS inhibited the TNF-α, IL-1β, IL-6, and IL-8 production in dose dependent manner. Maximal inhibition rate of the TNF-α, IL-1β, IL-6, and IL-8 by SHJGS (1.0 mg/ml) was 68.01 ± 0.28% (P < 0.01), 52.11 ± 0.56 % (P < 0.01), 53.42 ± 0.46 % (P < 0.01), and 46.70 ± 0.37% (P < 0.05), respectively. In addition, we show that SHJKS suppressed nuclear factor (NF)-κB activation induced by LPS plus PHA, leading to suppression of IκB-α phosphorylation and degradation. These results suggest that SHJKS might have regulatory effects on LPS plus PHA-induced cytokine production and NF-κB activation, which might explain its beneficial effect in the treatment of CI.     

妊娠期高血压疾病胎盘组织中CASR和EGFR的相关研究

目的探讨在妊娠期高血压疾病患者胎盘组织中细胞外钙受体（CASR）和表皮生长因子受体（EGFR）的表达情况及其相互关系。方法通过免疫组织化学方法检测妊娠期高血压疾病患者64例（妊娠期高血压组21例,子痫前期轻度组23例,重度组20例）及健康足月孕妇20例（对照组）胎盘组织中CASR和EGFR蛋白的表达情况。结果 （1）妊娠期高血压疾病患者胎盘组织中CASR在妊娠期高血压组表达水平为59.0532±8.039,子痫前期轻度组患者中为64.3623±3.7278,两组比较,差异有统计学意义（P〈0.0001）;CASR在子痫前期重度组患者中表达为112.2831±6.2060,与妊娠期高血压组比较,差异有统计学意义（P〈0.0001）,与子痫前期轻度组比较,差异有统计学意义（P〈0.0001）。CASR的蛋白在妊娠期高血压组患者胎盘组织中表达水平为59.0532±8.039,对照组为54.8585±4.3035,两组比较,差异无显著性（t=0.08,P=0.7759）。（2）妊娠期高血压疾病患者胎盘组织中表皮生长因子受体（EGFR）在妊娠期高血压组表达水平为56.174±3.1020,子痫前期轻度组患者中为78.6844±2.6713,两组比较,差异有统计学意义（t=18.73,P=0.0001）;EGFR在子痫前期重度组患者中表达为94.2090±6.8352,与子痫前期轻度组比较,差异有统计学意义（t=11.37,P〈0.0001）。（3）胎盘组织中CASR和表皮生长因子受体（EGFR）表达量呈正相关关系（r=0.352,P〈0.05）。结论妊娠期高血压疾病患者胎盘组织中CASR的蛋白表达升高和EGFR激活及过度增高,可能在妊娠期高血压疾病发生发展中起重要作用。     

Midtrimester Amniotic Fluid Tumor Necrosis Factor-Alpha Does Not Predict Small-for-Gestational-Age Infants

PROBLEM: To evaluate the independent ability of midtrimester amniotic fluid tumor necrosis factor-alpha (TNF-α) in the prediction of small-for-gestational-age (SGA) infants. METHOD OF STUDY: In this case-control study, patients delivering a SGA infant were matched with controls based on GA at delivery, maternal age, race, and parity. Patients with immune disease, chronic hypertension, diabetes, asthma, congenital hearts disease, multiple gestation, and fetal anomalies were excluded. Amniotic fluid samples were immunoassayed for TNF-α. Potential confounding variables evaluated were maternal serum alpha-fetoprotein level, smoking history, pregnancy induced hypertension, and neonatal gender. Statistical analysis included Fisher's exact test and ANOVA after log transformation with P < 0.05 considered significant. RESULTS: Eighteen patients delivered SGA neonates and were matched with 41 controls. No significant differences were identified in the confounding variables between patients with SGA neonates and controls. Amniotic fluid TNF-α levels were not significantly different between patients subsequently delivering SGA neonates and controls [median 7.63 (range 0.25-16.1) pg/mL versus 9.39 (0.25–66.9) pg/mL, P = 0.8]. CONCLUSIONS: Midtrimester amniotic fluid TNF-α levels are not predictive of SGA neonates when compared with controls matched for gestational age at delivery.     

Inhibition of pro-inflammatory cytokines and inducible nitric oxide by extract of Emilia sonchifolia L. aerial parts

Emilia sonchifolia L. (Asteraceae) is used in ethnomedicine for the treatment of a wide array of inflammatory disorders. This practice has also been supported by scientific reports which showed that extracts of E. sonchifolia possess anti-inflammatory effects in rodents. However, the mechanism(s) through which the extracts produce these effects is not known. In this study, the effect of a methanol/methylene chloride extract of E. sonchifolia (ES) on the levels of IL-1β and TNF-α after an intraperitoneal lipopolysaccharide (LPS; 1?mg/kg) challenge was investigated in mice. The effect of ES on TNF-α and inducible nitric oxide (iNO) production by LPS-stimulated bone marrow-derived macrophages (BMMDM) was also investigated in vitro. BMMDM were pre-incubated for 2?h with ES (20, and 100 μg/mL) or with Pyrrolidine dithiocarbamate, PDTC (100 µM) and then activated with LPS, and then the IL-1β, TNF-α and NO production measured in the cell-free conditioned culture supernatant after 24?h of incubation. In groups of mice pre-treated with ES, the systemic levels of IL-1β and TNF-α induced by LPS were found to be significantly (p < 0.05) lower. In vitro, ES treatment caused a concentration-dependent decrease in LPS-inducible IL-1β, TNF-α, and NO production by BMDM compared to the effects of treatment of the cells with LPS alone without affecting the viability of the cells. The results of these studies suggest that treatment with ES alleviated inflammatory responses possibly through a suppression of pro-inflammatory mediators and cytokines such as IL-1β, TNF-α, and iNO.     

Analysis of peripheral blood T-cell subsets,natural killer cells and serum levels of cytokines in children with Down syndrome

The objective of this study was to evaluate the relationship between humoral and cell-mediated immune response parameters and impairment of immune functions in children with Down syndrome (DS). The patient group was consisted of cytogenetically documented 32 children with DS. Lymphocyte subsets and natural killer cells were counted by flow-cytometry system. Levels of interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-8, IL-10 and tumour necrosis factor-alpha (TNF-α) were detected by enzyme-linked immunosorbent assay method. Serum IgG, IgM, IgA levels were measured by turbidimetric methods. The percentage of CD8+ lymphocytes and CD56+ cells of patients with DS were significantly higher, whereas CD20+ lymphocytes were lower than that of controls (P < 0.05). The percentage of CD2 and CD4 levels and CD4/CD8 ratio of patients with DS and normal controls were similar (P > 0.05). Levels of IL-4 and IL-10 were significantly increased, but IL-6 and TNF-α levels were decreased in children with DS (P < 0.05). Levels of other studied cytokines between patients with DS and controls were not statistically different (P > 0.05, for all). Serum IgG, IgM and IgA levels were found to be similar between the groups (P > 0.05). It has been known that IL-4 and IL-10 are anti-inflammatory molecules which inhibit the synthesis of proinflammatory cytokines such as IL-6 and TNF-α. In this study, levels of IL-4 and IL-10 were significantly increased, but IL-6 and TNF-α levels were decreased in children with DS. These results may suggest that continuing anti-inflammatory state in DS and this process may explain the cause of recurrent infection of the disease. On the other hand, in contrast to the low percentage of CD20+ cells, high percentage of CD8+ and CD56+ cells were found. Our findings may demonstrate that the cell-mediated and humoral immune system parameters in children with DS were altered according to healthy children.     

胃溃疡患者血清IL-6、TNF-α和TGF-β1检测的临床评价

目的：探讨胃溃疡患者治疗前后血清白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和转化生长因子-β1（TGF-β1）检测的临床意义。方法：应用放射免疫分析对33例胃溃疡患者进行治疗前后血清IL-6、TNF-α和TGF-β1检测,并与35位正常健康人作比较。结果：胃溃疡患者在治疗前血清IL-6、TNF-α和TGF-β1水平显著地高于正常人组（P〈0.01）,经中西医结合治疗3个月后则与正常人组比较无显著性差异（P〉0.05）。结论：检测胃溃疡患者治疗前后血清IL-6、TNF-α和TGF-β1水平的变化可提供对了解患者的病情和预后,因而具有重要的临床价值。     

子痫患者血清TNF-α、IL-6、 CRP、 Lp-PLA2、PON-1和APN水平的变化及临床研究

目的 探讨肿瘤坏死因子(TNF-α)、白介素6(IL-6)、C反应蛋白(CRP)、血清脂蛋白相关磷脂酶A2(Lp-PLA2)、对氧磷酶1(PON-1)以及脂联素(APN)在子痫前期孕妇血清中的变化及其临床意义.方法 选择2013年8月至2016年6月我院产科进行检测的孕妇160例,其中,妊娠期高血压孕妇(GH),轻度子痫前期孕妇(MPE)、重度子痫前期孕妇(SPE)和正常妊娠期孕妇(NC)各40例.采用酶联免疫吸附法检测IL-6、TNF-α以及Lp-PLA2水平,采用乳胶增强透射免疫比浊法检测血清CRP含量,利用qRT-PCR法检测PON-1和APN mRNA表达水平,采用免疫印迹法检测PON-1和APN蛋白表达水平,并比较各组之间的差异.结果 子痫前期孕妇血清TNF-a、IL-6、CRP以及Lp-PLA2水平明显高于正常妊娠对照组及妊娠期高血压组,差异存在统计学意义(P＜0.05).重度子痫前期患者血清TNF-a、IL-6、CRP以及Lp-PLA2水平明显高于轻度子痫前期患者,差异存在统计学意义(P＜0.05).子痫前期孕妇血清PON-1和APN mRNA和蛋白表达水平明显低于正常妊娠对照组及妊娠期高血压组,差异存在统计学意义(P＜0.05),且重度子痫前期患者血清PON-1和APNmRNA和蛋白水平明显低于轻度子痫前期患者,差异存在统计学意义(P＜0.05).结论 子痫前期患者血清中TNF-a、IL-6 、CRP以及Lp-PLA2水平的升高,PON-1和APN水平的降低可能参与子痫前期的发生.     

Maternal Levels of Prostacyclin,Thromboxane, ICAM,and VCAM in Normal and Preeclamptic Pregnancies

Citation Lewis DF., Canzoneri BJ., Gu Y, Zhao S, Wang Y. Maternal Levels of Prostacyclin, Thromboxane, ICAM, and VCAM in normal and preeclamptic pregnancies. Am J Reprod Immunol 2010; 64: 376–383 Problem To evaluate whether impaired endothelial function and endothelial inflammatory response occur in parallel in the women with preeclampsia. Method of Study Venous blood was drawn from normal (n = 40) and severe preeclamptic (sPE) (n = 40) pregnant women when they were admitted to the L&D Unit and 24 hrs after delivery. Plasma and serum samples were extracted and measured for 6‐keto PGF1α and TXB₂ (stable metabolites of PGI2 and TXA2), and intercellular adhesion molecule (ICAM) and vascular cell adhesion molecule (VCAM) by ELISA. Data are analyzed by Mann–Whitney test and paired t‐test. The statistical significance is set as P < 0.05. Results Plasma 6‐keto PGF1α levels were significantly reduced at admission and 24 hr after delivery in sPE compared to normal pregnant controls, P < 0.01. The ratio of 6‐keto PGF1α and TXB₂ was significant less in sPE than that in normal pregnant controls before delivery. There was no significant difference for ICAM and VCAM levels between normal and patients with sPE before and after delivery. Conclusion Maternal 6‐keto PGF1α levels and the ratio of 6‐keto PGF1α and TXB₂ were decreased in patients with sPE compared to normal pregnant controls. In contrast, maternal ICAM and VCAM levels were not different between the two groups. These data suggest that serum ICAM and VCAM levels may not be sensitive inflammatory biomarkers for preeclampsia.     

Association of longevity with IL-10 −1082 G/A and TNF-α−308 G/A polymorphisms

Cytokines are crucial for the regulation of inflammation development in humans. Many studies have shown that variations in cytokine genes might play a role in determining human longevity. This study examined the changes in the gene pool relevant to the −308 G/A polymorphism in the promoter region of the proinflammatory cytokine tumour necrosis factor (TNF)-α gene and the −1082 G/A polymorphism in the promoter region of anti-inflammatory cytokine interleukin (IL)-10 gene with aging and survival selection occurs in the Jordanian population. IL-10 −1028 G/A and TNF-α−308 G/A were genotyped in 119 randomly selected elderly subjects (41 women and 78 men) with a mean age of 90.2 years and young control subjects of 118 (46 women and 72 men) with a mean age of 31.9 years. No significant differences were found in the genotype and allele frequencies of TNF-α gene variants between the two groups (P > 0.05) while the IL-10 genotype and allele frequencies were significantly associated with longevity in men (P < 0.05) but not in women (P < 0.05). Thus, IL-10 −1028 G/A polymorphism seems to play a role in the pathway to longevity in Jordanian men.     

MMP-9和IL-12在妊娠期高血压疾病胎盘上的表达

目的通过分析子痫前期患者与正常晚孕妇女胎盘组织中基质金属蛋白酶一9和白介素12的表达,探讨其在妊娠期高血压疾病(PIN)中的作用及两者的相关性。方法采用免疫组织化学染色法测定MMP-9和IL-12在28例妊娠期高血压疾病患者(10例为子痫前期轻度;18例为子痫前期重度)和10例正常妊娠患者胎盘上的表达。结果妊娠期高血压疾病组胎盘MMP-9的表达明显低于正常妊娠组(P<0.01);妊娠高血压疾病组胎盘IL-12的表达明显高于正常妊娠组(P>0.01),随病情加重MMP-9的表达有下降趋势;IL-12的表达有升高趋势,有统计学意义。结论与正常妊娠相比妊娠期高血压疾病组胎盘IL-12表达明显增加,MMP-9表达明显下降,且两因子表达与病情正相关。表明两因子参与了妊娠期高血压疾病的发生与发展。     

Gonadotropins Induce the Release of Interleukin-1β, Interleukin-6 and Tumor Necrosis Factor-α from the Human Preovulatory Follicle

PROBLEM; The effects of exogenous gonadotropin administration and steroid levels on the release of various cytokines into the human follicular fluid (FF) were studied. METHOD OF STUDY: Forty patients were included in two groups, those undergoing controlled ovarian hyperstimulation (COH) (n = 33) and natural cycles (n = 7). FF transvaginal aspirations were performed 36 hr after administration of human chorionic gonadotropin or a spontaneous surge of luteinizing hormone, respectively. FF cytokine measurements were performed with sensitive immunoassays. RESULTS: FF cytokine levels were higher after COH [interleukin (IL)-1β, 6.6 ± 0.32 pg/ml; IL-6, 18.7 ± 2.1 pg/ml; and tumor necrosis factor (TNF)-α, 32.5 ± 4.9 pg/ml] than in natural unstimulated cycles (0.52 ± 0.1 pg/ml, P < 0.001; 8.9 ± 1.2 pg/ml, P < 0.01; and 13.2 ± 2.6 pg/ml, P < 0.001, respectively). FF estradiol (E₂) and progesterone levels were not statistically different between groups, despite the higher serum E₂ levels observed in patients after COH. CONCLUSIONS: Gonadotropins might regulate ovarian secretion of cytokines, because FF IL-1β, IL-6, and TNF-α levels after COH were higher than during natural cycles.     

Quantitative intracellular cytokine measurement: age-related changes in proinflammatory cytokine production

The proinflammatory cytokines play a central role in mediating cellular and physiological responses, and levels may reflect immune system effectiveness. In this study, the effect of ageing on the inflammatory response was examined using a novel method to detect production of the proinflammatory cytokines, i.e. tumour necrosis factor-alpha (TNF-α), IL-6 and IL-1β. Peripheral blood mononuclear cells (PBMC) obtained from healthy donors of different ages were incubated for 0, 24, 48 and 72 h with or without phorbol 12-myristate 13-acetate (PMA) stimulation. At each time point these cells were permeabilized and incubated with secondary conjugated FITC MoAbs specific for each cytokine. A flow cytometric system was developed to quantify specific intracellular fluorescence in T cells (CD3⁺) and monocytes (CD14⁺). TNF-α, IL-6 and IL-1β production in cell culture supernatants was also measured using ELISAs. In older subjects, flow cytometry detected significant increases in intracellular T cell TNF-α and IL-6 (P < 0.05). IL-1β was not detected in any of the T cell samples. Likewise, the monocytes of older subjects demonstrated increased intracellular levels of all three cytokines, but these increases were not significant (P > 0.05). These changes in intracellular proinflammatory cytokine levels may explain some of the exaggerated inflammatory responses seen in elderly patients.     

Evidence for an Elevation in Serum Interleukin-2 and Tumor Necrosis Factor-α Levels Before the Clinical Manifestations of Preeclampsia

PROBLEM: The purpose of this study is to clarify whether the disruption of immune regulation occurs in early pregnancy before the clinical manifestations of preeclampsia. METHOD OF STUDY: The serum concentrations of interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α) were determined by using enzyme-linked immunoadsorbent assay (ELISA) in the first trimester of pregnancy in women who had preeclampsia develop after 28 weeks of pregnancy (preeclamptic group) and in women who completed pregnancy uneventfully (control group). RESULTS: Serum concentrations of both IL-2 and TNF-α in the first trimester of the preeclamptic group were significantly higher than those of the control group. CONCLUSIONS: That the perturbation of feto-maternal immune regulation may precede the clinical manifestations of preeclampsia, which may be of relevance in the development of preeclampsia, is suggested.