Interleukin-1 receptor antagonist gene polymorphism,vaginal interleukin-1 receptor antagonist concentrations,and vaginal ureaplasma urealyticum colonization in pregnant women

Ureaplasma urealyticum is the microorganism most frequently isolated from amniotic fluids of women in preterm labor. The relationship between vaginal colonization with U. urealyticum, vaginal interleukin-1 receptor antagonist (IL-1ra) levels, and the IL-1ra genotype in pregnant women was examined. Vaginal specimens, obtained with a cotton swab from 207 women in their first trimester of pregnancy, were tested for IL-1ra concentrations by enzyme-linked immunosorbent assay and for U. urealyticum and IL-1ra genotypes by PCR. U. urealyticum was detected in 85 (41.1%) women. The median IL-1ra level was 450 ng/ml in women positive for U. urealyticum, as opposed to 225 ng/ml in women negative for this microorganism (P < 0.0001). Sixty-two percent of the 16 women who were homozygous for allele 2 of the IL-1ra gene (IL-1RN*2) were colonized with U. urealyticum, as opposed to 47% of the 49 women who were IL-1RN*1/IL-1RN*2 heterozygotes and 34% of the 133 women who were IL-1RN*1 homozygotes (P < 0.05). Median IL-1ra levels were 750 ng/ml in IL-1RN*2 homozygotes, 300 ng/ml in IL-1RN*1/IL-1RN*2 heterozygotes, and 250 ng/ml in IL-1RN*1 homozygotes (P = 0.02). The vast majority of subjects had an uneventful pregnancy and delivered a healthy infant at term. The IL-1ra genotype or U. urealyticum colonization was unrelated to birth weight. Pregnant women who are colonized with U. urealyticum during the first trimester have elevated vaginal IL-1ra concentrations and a higher prevalence of the IL-1RN*2 homozygote genotype than do noncolonized women.     

Interleukin-1 receptor antagonist modulates inflammation and scarring after ligament injury

Ligaments have limited regenerative potential and as a consequence, repair is protracted and results in a mechanically inferior tissue more scar-like than native ligament. We previously reported that a single injection of interleukin-1 receptor antagonist (IL-1Ra) delivered at the time of injury, decreased the number of M2 macrophage-associated inflammatory cytokines. Based on these results, we hypothesized that IL-1Ra administered after injury and closer to peak inflammation (as would occur clinically), would more effectively decrease inflammation and thereby improve healing. Since IL-1Ra has a short half-life, we also investigated the effect of multiple injections. The objective of this study was to elucidate healing of a medial collateral ligament (MCL) with either a single IL-1Ra injection delivered one day after injury or with multiple injections of IL-1Ra on days 1, 2, 3, and 4. One day after MCL injury, rats received either single or multiple injections of IL-1Ra or PBS. Tissue was then collected at days 5 and 11. Both single and multiple IL-1Ra injections reduced inflammatory cytokines, but did not change mechanical behavior. A single injection of IL-1Ra also reduced the number of myofibroblasts and increased type I procollagen. Multiple IL-1Ra doses provided no additive response and, in fact, reduced the M2 macrophages. Based on these results, a single dose of IL-1Ra was better at reducing the MCL-derived inflammatory cytokines compared to multiple injections. The changes in type I procollagen and myofibroblasts further suggest a single injection of IL-1Ra enhanced repair of the ligament but not sufficiently to improve functional behavior.     

Interleukin-1 receptor antagonist gene polymorphism and mortality in patients with severe sepsis

This study aims to determine the influence of the polymorphism within the intron 2 of the interleukin-1 receptor antagonist gene (IL-1RN*) on the outcome of severe sepsis, and to assess its functional significance by correlating this polymorphism with the total production of interleukin-1 receptor antagonist (IL-1Ra) protein determined in stimulated peripheral blood mononuclear cells (PBMC). A group of 78 patients with severe sepsis (51 survivors and 27 nonsurvivors) was compared with a healthy control group of 130 blood donors, and 56 patients with uncomplicated pneumonia. We found a significant association between IL-1RN* polymorphism and survival. Thus, after adjusting for age and APACHE II score, multiple logistic regression analysis showed that patients homozygotes for the allele *2 had a 6.47-fold increased risk of death (95% CI 1.01--41.47, P = 0.04). Besides, compared with patients homozygous or heterozygous for the allele *1, IL-1RN*2 homozygotes produced significantly lower levels of IL-1Ra from their PBMC. Our results suggest that insufficient production of this cytokine might contribute, among other factors, to the higher mortality rate found in severe sepsis patients with the IL-1RN*2 homozygous genotype.     

Interleukin-1 receptor antagonist restores homeostatic function and limits multiorgan damage in heatstroke

We measured diffusing capacity (DLCO), alveolar membrane properties (D _m), capillary lung volume (V _c), and alveolar volume (V _A ) in 20 healthy subjects (12 males; age 32.4 ± 13 (SD); BMI 21.7 ± 3; non smokers) at total lung capacity (TLC) and at ∼80, 60, and 40% TLC. In all subjects, D _m increased with lung volume, the increase being significantly greater for higher values of D _m(TLC): the inter-individual differences can be interpreted by a greater number of alveolar units coupled to a lower thickness of the air–blood barrier (thus a higher alveolar surface to thickness ratio S _A/τ). On the average, the volume-dependent increase of D _m from ∼40 to 100% TLC is less than expected based on geometrical increase of S _A /τ. In fact, up to ∼80% TLC, the increase in D _m closely reflects only the increase of S _A, suggesting “unfolding” of the septa with no appreciable decrease in τ. Conversely, above 80% TLC, the decrease in τ due to parenchymal stretching becomes the main factor affecting D _m. In all subjects, V _c decreased with increasing lung volume, in line with an increase in parenchymal stretching; the decrease was significantly larger for higher values of V _c (40% TLC). Possibly reflecting differences in alveolar capillary density. No correlation was found between D _m(TLC) and V _c(40%TLC). The individual specificity in the lung volume dependence of V _c and D _m can be reasonably described by evaluating the V _c/D _m ratio at TLC and at ∼40%TLC.     

Interleukin-1beta and interleukin-1 receptor antagonist gene polymorphism in postmenopausal women: correlation to bone mineral density and susceptibility to osteoporosis

OBJECTIVE: Osteoporosis is a common disorder with a strong genetic component. Our aim was to investigate the correlations of the interleukin-1beta (IL-1beta) and interleukin-1 receptor antagonist (IL-1Ra) gene polymorphisms with bone mineral density (BMD) and their relationship to osteoporosis. METHODS: The IL-1beta (promoter and exon 5) and IL-1Ra (intron 2) gene polymorphisms were determined using polymerase chain reaction. BMD of the lumbar spine and proximal femur were measured using dual-energy X-ray absorptiometry. RESULTS: The prevalence of each genotype of the interleukin-1 related genes in the study population was: (1) 14% C/C, 71.5% C/T, and 14.5% T/T in IL-1beta promoter; (2) 95.3% E1/E1 and 4.7% E1/E2 in IL-1beta exon 5; (3) 92.4% I/I, 6.4% I/II, and 1.2% II/II in IL-1Ra intron 2. After adjustment for potential confounding factors such as age, height, weight, years since menopause, and daily calcium intake, subjects with genotype E1/E2 (n=8) in IL-1beta exon 5 had lower BMD values and a significantly greater risk for osteoporosis (OR 10.6, 95% CI 1.3-83.8) at the lumbar spine when compared with subjects with genotype E1/E1 (n=164) in IL-1beta exon 5. CONCLUSION: The Taq I IL-1beta exon 5 gene polymorphism is associated with reduced BMD and predisposes women to osteoporosis at the lumbar spine, but our results should be interpreted with caution because of the small number of subjects with the unfavorable E1/E2 genotype.     

TRPC1是人气道上皮细胞感受压力的受体

目的探讨机械敏感瞬时受体电位蛋白-1(TRPC1)在人气道上皮细胞感受压力过程中的作用。方法 1)收集人肺癌患者肺癌旁5 cm以外的无病变组织,根据临床表现分为正常组、慢性阻塞性肺病组(COPD)和哮喘组,通过细胞免疫组织化学、Western blot及实时荧光定量聚合酶链反应(qRT-PCR)检测人气道上皮细胞内TRPC1表达水平。2)TRPC1 siRNA、NC siRNA转染16HBE细胞,细胞免疫荧光技术检测基因沉默效率。3)将16HBE细胞分为对照组、单纯刺激组、刺激+TRPC1 siRNA转染组,以细胞牵张刺激系统给予刺激,激光共聚焦观察细胞内钙离子浓度变化。结果 1)COPD组、哮喘组TRPC1蛋白表达及转录水平明显高于对照组(P0.05)。2)TRPC1 siRNA转染组细胞内TRPC1荧光强度明显低于对照组(P0.05),NC siRNA组较正常组无明显差别。3)单纯刺激组细胞内钙离子荧光强度显著高于对照组(P0.05),而刺激+TRPC1 siRNA转染组钙离子荧光强度与对照组无明显差异。结论 TRPC1是人气道上皮细胞内重要的压力敏感受体。     

Interleukin-1 beta (IL-1 beta), IL-1 receptor antagonist, and TNF alpha production in whole blood.

The ability of an individual to mount defense responses to infection depend in part on the capacity to produce cytokines such as interleukin 1 (IL-1) and tumor necrosis factor (TNF). The specialized equipment, labor intensity, and sterile practice required for the standard in vitro evaluation of cytokine production can make such evaluation impractical in some clinical situations. We report a method for stimulating whole blood to produce cytokines that can be implemented in laboratories without tissue culture facilities and requires minimal sample preparation. IL-1 beta and TNF alpha production in whole blood samples was stimulated with endotoxin and/or phytohemagglutinin in standard EDTA-containing vacuum collection tubes. After incubation, plasma was removed and frozen for later assay. Comparison of this whole blood method with isolated mononuclear cell cultures indicated a significant correlation for IL-1 beta production (r = 0.746, P = 0.005). This technique also produced the newly described cytokine, IL-1 receptor antagonist. We conclude that the whole blood method is an acceptable alternative to isolated cell culture methods for measuring IL-1 beta in situations that preclude the standard in vitro approach.     

Association of interleukin-1beta and interleukin-1 receptor antagonist polymorphisms with bacterial vaginosis in non-pregnant Italian women

Bacterial vaginosis (BV) is the most prevalent alteration of vaginal microflora worldwide. BV is a polymicrobial disorder, and its etiology is elusive. Factors predisposing to this recurrent condition are not fully characterized. We aimed to investigate whether interleukin-1beta (IL-1beta) and IL-1 receptor antagonist (IL-1ra) polymorphisms are associated with BV in non-pregnant white Italian women. Genomic DNA was obtained from 164 BV positive, and 406 control women. Two diallelic polymorphisms in the IL-1beta gene (IL-1B) representing C/T base transitions at - 511 and + 3954 positions and a variable number tandem repeats (VNTR) in intron 2 of the IL-1ra gene (IL-1RN) were assessed. We demonstrated that women who were homozygous for - 511 CC or + 3954 TT of the IL-1B gene were at increased risk for BV with an odds ratio (OR) = 1.5 [95% confidence interval (CI) = 1.03-2.14, P = 0.032], and OR = 2.8 (95% CI = 1.37-5.88, P = 0.004), respectively. The haplotype - 511/ + 3954 T-C was protective for BV, with an OR = 0.7 (95% CI = 0.49-0.90, P = 0.009). The IL-1RN VNTR genotype was not associated with BV, although the rare allele 3 showed a trend towards protection (P = 0.049). These data show that host genetic variants at the IL-1beta locus predispose to BV among Caucasian non-pregnant women. Further studies will determine whether these genetic polymorphisms modulate the risk for BV recurrence, and/or BV associated severe adverse outcomes as preterm birth and human immunodeficiency virus transmission.     

Interleukin-1 receptor antagonist in inflammatory exudate cells of rabbits. Production, purification and determination of primary structure

A rabbit interleukin-1 (IL-1) inhibitor in inflammatory peritoneal exudate cells was purified to apparent homogeneity. This inhibitor was extracted from exudate cells of the 24-hr stage of casein-induced peritoneal inflammation and purified using isoelectrofocusing (IEF), gel filtration, followed in this order by high-performance liquid chromatography (HPLC) steps with hydroxylapatite and anionic ion exchanger. The purified factor showed a single band on silver-stained SDS-PAGE. This molecule of MW 19,000 and pI 5.5 inhibited the binding of both IL-1 alpha and beta to receptors on a thymoma cell line, EL-4 and a B-cell line, 70Z/3. We determined its primary structure by a combination of peptide chemistry and molecular cloning. The inhibitor was synthesized as a precursor composed of 177 amino acids and was processed to a mature molecule of 143 amino acids. The N-terminal amino acid of the mature inhibitor was N-acetyl-methionine residue. The deduced amino acid sequence of the inhibitor showed a 77% homology to the human IL-1 receptor antagonist (IL-1Ra) and essentially the same mode of action as seen with human IL-1Ra. We consider that this inhibitor is a rabbit counterpart of human IL-1Ra, although there are differences with respect to the molecular structure; the N-terminus of the mature rabbit IL-1Ra at a position of nine amino acids downstream from that of human IL-1Ra.     

Interleukin-1 and cancer progression: the emerging role of interleukin-1 receptor antagonist as a novel therapeutic agent in cancer treatment

The tumor microenvironment consists of tumor, immune, stromal, and inflammatory cells which produce cytokines, growth factors, and adhesion molecules that promote tumor progression and metastasis. Of particular interest in this setting is interleukin-1 (IL-1), a pleiotropic cytokine with numerous roles in both physiological and pathological states. It is known to be up regulated in many tumor types and has been implicated as a factor in tumor progression via the expression of metastatic and angiogenic genes and growth factors. A number of studies have reported that high IL-1 concentrations within the tumor microenvironment are associated with a more virulent tumor phenotype. Solid tumors in which IL-1 has been shown to be up regulated include breast, colon, lung, head and neck cancers, and melanomas, and patients with IL-1 producing tumors have generally bad prognoses. The exact mechanisms by which IL-1 promotes tumor growth remain unclear, though the protein is believed to act via induction of pro-metastatic genes such as matrix metalloproteinases and through the stimulation of adjacent cells to produce angiogenic proteins and growth factors such as VEGF, IL-8, IL-6, TNFα, and TGFβ. The IL-1 receptor antagonist (IL-1ra) is a naturally occurring inhibitor to IL-1 and acts by binding to the IL-1 receptor without activating it. The protein has been shown to decrease tumor growth, angiogenesis, and metastases in murine xenograft models. Our focus in this review is to summarize the known data on the role of IL-1 in tumor progression and metastasis and the use of IL-1 inhibition as a novel therapeutic approach in the treatment of solid organ malignancies.     

Elevated concentrations of interleukin-1 beta and interleukin-1 receptor antagonist in plasma of women with silicone breast implants.

Plasma from 27 women with silicone breast implants (SBIs) and 50 age-matched control women without SBIs were examined by enzyme immunoassay for the presence of interleukin-1 beta (IL-1 beta) and its naturally occurring receptor antagonist, IL-1ra. The results show that 74% (20 of 27) of women with SBIs had elevated concentrations of IL-1ra, whereas only 2% (1 of 50) of controls without SBIs had elevated concentrations of IL-1ra. In contrast to the IL-1ra results, the frequency of elevated IL-1 beta concentrations among women with SBIs was only 40% (11 of 27), but this was significantly higher than the 0% (0 of 50) in control women without SBIs. These findings suggest that there is a chronic ongoing inflammatory process in some women with SBIs, the implications of which are discussed in the context of silicone as an antigenic stimulant of the immune system.     

Interleukin-1 (IL-1) receptor antagonist prevents Staphylococcus epidermidis-induced hypotension and reduces circulating levels of tumor necrosis factor and IL-1 beta in rabbits.

Similar to shock in gram-negative sepsis, shock from gram-positive organisms is mediated, in part, by tumor necrosis factor (TNF) and interleukin-1 (IL-1). In the present study, rabbits were infused with IL-1 receptor antagonist (IL-1ra) prior to and during Staphylococcus epidermidis-induced hypotension. After injection of bacteria, a maximal fall in mean arterial pressure to -42% below baseline occurred at 200 min in vehicle-treated animals compared with a nonsignificant decrease of only 7% in the IL-1ra-treated group (P < 0.01, vehicle versus IL-1ra). A similar attenuation was observed in the fall in systemic vascular resistance (P < 0.05). After the injection of S. epidermidis, TNF levels rose to a peak elevation of 475 +/- 160 U/ml in vehicle-treated rabbits, but in rabbits receiving IL-1ra, maximal TNF levels rose only to 85 +/- 23 U/ml (P < 0.01). Plasma IL-1 beta reached maximal concentrations at 180 min of 364 +/- 71 pg/ml in vehicle-treated animals but only 145 +/- 12 pg/ml in rabbits given IL-1ra (P < 0.05). The reductions in TNF and IL-1 were not due to interference by IL-1ra in the respective assays. In vitro, IL-1ra inhibited S. epidermidis-induced TNF from mononuclear cells by 31% +/- 11%, from spleen cells by 17% +/- 4% (P < 0.05), and from whole blood by 42% +/- 17%. Despite the near reversal of the fall in mean arterial pressure and systemic vascular resistance in IL-1ra-treated rabbits, leukopenia and thrombocytopenia were unaffected. These results demonstrate that IL-1ra blocks shock-like hemodynamic parameters and reduces circulating IL-1 and TNF levels in a model of gram-positive sepsis.     

Effect of alpha1-adrenergic receptor antagonist on the noradrenaline-induced facilitation in respiratory rhythm in newborn rat pons-medulla-spinal cord preparations

We hypothesized that facilitation of respiratory rhythm by noradrenaline (NA) in rat pons-medulla-spinal cord preparations is mediated through alpha1-adrenergic receptors. In 0- to 4-day-old rats, the respiratory frequency (fR) was monitored at the C4 ventral root and trigeminal motor (VMO) outputs. fR at temperature (Te)=23 degrees C was lower than that at a higher Te (27 degrees C) and was increased by NA. At 23 degrees C, lower concentrations of NA were needed to produce the same increases in fR seen at 27 degrees C. With highest NA concentration we tested (50 microM), activity at C4 was maintained in all preparations at both Te, whereas that at VMO was maintained in 50% (27 degrees C) or 88% (23 degrees C) of the preparations. Particularly, tonic activity at C4 appeared in all preparations at both Te, but that at the VMO occurred in 0% (27 degrees C) or 18% (23 degrees C) of the preparations. Based on these results, we used the lower Te (23 degrees C) and applied a low concentration of NA (3 microM) to the preparations. We found that: (1) with the addition of NA, fR was increased without the occurrence of tonic activity and (2) NA-related fR facilitation was inhibited by pre-treatment with the alpha1-adrenergic receptor antagonist prazosin (2 microM). fR was increased by application of the alpha1-adrenergic receptor agonist phenylephrine (4 microM), and this response was inhibited by prazosin (4 microM). At Te=23 degrees C, fR facilitation by NA in newborn rat pons-medulla-spinal cord preparations was obtained by activation of alpha1-adrenergic receptors.     

Nonselective cation channels are essential for maintaining intracellular Ca2+ levels and spontaneous firing activity in the midbrain dopamine neurons

Intracellular Ca²⁺ and Ca²⁺-permeable ion channels are important in regulating the firing activity and pattern of midbrain dopamine neurons, but the role of Ca²⁺-permeable nonselective cation channels (NSCCs) on spontaneous firing activity is unclear. Therefore, we investigated how Ca²⁺-permeable NSCCs modulate spontaneous firing activity and cytosolic Ca²⁺ concentration ([Ca²⁺]_c) in acutely isolated midbrain dopamine neurons of the rat. Applications of voltage-dependent Ca²⁺ channels antagonists failed to abolish spontaneous firing activity completely, but they decreased firing rate and [Ca²⁺]_c. However, a blockade of NSCCs by 2-APB or SKF96365 more potently suppressed spontaneous firings with a depolarization of membrane potential and strong decreases in basal [Ca²⁺]_c levels. The depolarization of membrane potentials was attenuated by intracellular dialysis with 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA). NSCCs blockers inhibited oscillatory potentials and decreased basal [Ca²⁺]_c in the presence of tetrodotoxin. Apamin, a small-conductance Ca²⁺-activated K⁺ channel inhibitor, depolarized membrane potentials and enhanced firing rates. From these data, we conclude that NSCCs not only make up the tonic Ca²⁺ entry pathways to uphold basal [Ca²⁺]_c levels but also contribute to generation of spontaneous firings, thereby regulating spontaneous firing activities of the midbrain dopamine neurons.     

Montelukast, a leukotriene receptor antagonist, inhibits the late airway response to antigen, airway eosinophilia, and IL-5-expressing cells in Brown Norway rats

BACKGROUND: Asthma is characterized by airflow obstruction, inflammatory cell infiltration, and the synthesis of mediators, such as T(H2) cytokines and leukotrienes, in the airways. Cysteinyl leukotriene (cysLT) receptor antagonists have recently been associated with clinical improvement of asthma and reduced airway inflammation. Whether the beneficial effects of cysLT antagonists are mediated through the modulation of cytokine expression has not been determined. OBJECTIVE: The aim of the study was to determine the presence of eosinophils and IL-5 messenger (m)RNA(+) cells within the lungs of antigen-challenged Brown Norway rats after treatment with the cysLT(1) receptor antagonist montelukast (MK). METHODS: Ovalbumin-sensitized Brown Norway rats were treated with either MK or saline before ovalbumin challenge. Pulmonary mechanics were monitored for 8 hours. Subsequently, immunocytochemistry and in situ hybridization were used to examine bronchoalveolar lavage (BAL) fluid and lung tissue for cells expressing major basic protein (eosinophils) and IL-5 mRNA, respectively. Simultaneous in situ hybridization and immunocytochemistry was used to phenotype the cells expressing mRNA encoding IL-5. RESULTS: Animals treated with MK had significantly lower lung resistance and fewer eosinophils and IL-5 mRNA(+) cells within BAL fluid and lung tissue compared with that found in saline-treated animals. Colocalizaton studies revealed that the majority of IL-5 mRNA(+) cells were T cells and that the number of IL-5 mRNA(+)/CD3(+) or IL-5 mRNA(+)/major basic protein(+) cells were significantly less within BAL from animals treated with MK than from those treated with saline. CONCLUSIONS: These results indicate that the cysLT(1) receptor antagonist MK can diminish the pulmonary response to antigen, tissue eosinophilia, and the number of cells expressing IL-5 mRNA, suggesting that leukotrienes may also regulate the allergic response through the modulation of inflammation and cytokine synthesis.     

Trophoblastic beta1-glycoprotein and hemostasis system in pregnant women with antiphospholipid syndrome

Clinical and laboratory studies were carried out in 38 pregnant women with antiphospholipid syndrome. Increased functional activity of platelets and decreased protein-producing function of the placenta were observed starting from the early terms of gestation. These disorders were followed by the development of hypercoagulation in the plasma component of hemostasis, appearance of intravascular blood clotting markers, and inhibition of AT III and protein C. This led to the progress of disorders in the microcirculatory bed, fetoplacental insufficiency, decrease in trophoblastic ₁-glycoprotein level, chronic hypoxia, and fetal death. Infection accelerated this process. Measurements of trophoblastic ₁-glycoprotein every 2 weeks help to diagnose fetoplacental disorders, predict the course of pregnancy, and evaluate the efficiency of drug therapy.     

Maternal vascular endothelial growth factor receptor and interleukin levels in pregnant women with twin-twin transfusion syndrome

Twin-twin transfusion syndrome (TTTS) is an unusual and serious condition that occurs in twin pregnancies when identical twins share a placenta but develop discordant amniotic fluid volumes. TTTS is associated with an increased risk of fetal death and birth defects if untreated. This study investigated the soluble levels of biomarkers including growth factors and interleukins in pregnant women with and without TTTS during pregnancy. We quantified plasma levels of VEGF-R1, VEGF-R2, IL-1β, IL-6 and IL-8 in twin pregnant women with (n=53) and without TTTS (n=72) and in women with single pregnancy (n=30) by ELISA and analyzed the association of maternal circulating biomarker levels with TTTS. Our results showed that maternal VEGF-R1 levels were significantly higher in twins compared to single pregnancy (P<0.05) and were decreased in the second trimester compared to the first trimester (P = 0.065, 0.019 and 0.072 for twins with and without TTTS and single pregnancy, respectively). VEGF-R2 levels had a trend to be lower in twins compared to single pregnancy. In addition, soluble VEGF-R1 and VEGF-R2 levels were significantly decreased while IL-6 levels were increased after surgical treatment with laser in twin pregnant women with TTTS (P = 0.016, 0.041 and 0.04, respectively). These results suggest that IL-6, VEGF-R1 and VEGF-R2 are involved in vascular regulation and stabilization in twin pregnancies and may contribute to the pathogenesis of TTTS and thus play a prognostic role in the surgical treatment of TTTS.     

人白介素1受体拮抗剂的cDNA克隆和原核表达

从活化的正常人外周血单核细胞中提取总ＲＮＡ，经逆转录多聚酶链反应（ＲＴ－ＰＣＲ）获得了人白介素１受体拮抗剂（ＩＬ－１Ｒａ）ｃＤＮＡ。经过ＤＮＡ测序分析，发现该片段和国外发表的ＩＬ－１ＲａｃＤＮＡ序列一致，将该目的基因插入ｐＢＶ２２０载体，并转入大肠杆菌ＨＢ１０１中，经热诱导表达重组蛋白，ＳＤＳ－ＰＡＧＥ后发现，菌体超声裂解后，在可溶性上清中有一Ｍ１７０００的特异带，约占菌体可溶性总蛋白的８０％以上     

Interleukin-6 and soluble interleukin-6 receptor serum levels in recurrent spontaneous abortion women immunized with paternal white cells

PROBLEM: Recurrent spontaneous abortion (RSA) could be interpreted as the cause for the incapacity of the mother to recognize paternal antigens to produce the desired protective response. The practise of alloimmunization was introduced in an attempt to induce in the mother the production of an alloimmune response; some authors proposed an association between cytokines and RSA. The production of IL6 and its soluble receptor (sIL6R) before and after lymphocyte immunotherapy was evaluated in sera of 33 patients suffering from two or more RSA, and in sera of 47 women with normal pregnancy. METHOD OF STUDY: The immunization of RSA patients was achieved by injection of four doses of 10(5) mononuclear cells (MNC) from the husband, at weekly intervals, before pregnancy. The IL6 and sIL6R levels were measured using sandwich ELISAs and the results evaluated by Tukey-Kramer multiple comparison-tests. RESULTS: Our data show no significant differences between IL6 and sIL6R serum levels of normal pregnant women and RSA pregnant women with white-cell immunization before pregnancy. In contrast, the sera of pregnant RSA patients without allogeneic therapy show higher values. We also found significant differences between IL6 levels in non-pregnant RSA women with and without immunotherapy. CONCLUSION: These results show that the alloimmunization with paternal white cells leads the serum IL6 and sIL6R-levels to the values observed in the course of normal pregnancy, suggesting a role for IL6 and sIL6R in the modulation of the immune response's quality.     

Production of IL-1 and IL-1 receptor antagonist and the pathological significance in lipopolysaccharide-induced arthritis in rabbits.

Injection of lipopolysaccharide (LPS) into rabbit knee joints provoked leucocyte infiltration and loss of proteoglycan (PG) from the cartilage. We investigated the role of IL-1 and IL-1 receptor antagonist (IL-1Ra) and its significance in the pathogenesis of LPS-arthritis. Production of IL-1 beta peaked at 6 h (196.7 +/- 89.4 pg/joint) after injection of 10 ng of LPS, while IL-1Ra peaked at 9 h (34.5 +/- 13.4 ng/joint). The amount of IL-1Ra was 180-200-fold molar excess of IL-1, and a large amount of IL-1Ra was sustained for 1 week. Both IL-1 beta and IL-1Ra were mainly produced by synovial exudate cells. Arthritis was reproduced by rabbit IL-1 beta. LPS-induced leucocyte infiltration was inhibited 70-75% by rabbit IL-1Ra. Loss of PG in LPS-arthritis was prevented by IL-1Ra and also by neutrophil elastase inhibitor, and superoxide dismutase. In leucopenic rabbits, injection of LPS induced neither production of IL-1 beta nor loss of PG. Direct injection of inflammatory exudated cells in leucopenic rabbits reproduced loss of PG, and there was only a partial recovery by IL-1Ra. These results suggest that LPS-initiated IL-1 acts as a key mediator in LPS-arthritis and that endogenous IL-1Ra may suppress a part of IL-1 activity at the site, but its amount was too low for suppression of the produced IL-1. Loss of PG is a sequela of infiltrated leucocytes and leucocyte-derived elastase, and superoxide anion may play a pivotal role in the destruction of cartilage.