Elevated interleukin-8 in cervical mucus as an indicator for treatment to prevent premature birth and preterm, pre-labor rupture of membranes: a prospective study

PROBLEM: We investigated whether cervical shortening and high interleukin (IL)-8 in cervical mucus were valuable indications for treatment to prevent premature birth and preterm, pre-labor rupture of membranes (pPROM). METHOD OF STUDY: Pregnant women were divided into group A, in which neither cervical IL-8 nor cervical length was measured in the middle trimester; and groups B and C, in which cervical length and cervical IL-8 were measured, and bed rest or cerclage was performed when cervical shortening was detected. In group B, vaginal washing with povidone iodine and insertion of chloramphenicol vaginal tablets were carried out in women with IL-8 elevations. RESULTS: In group B, duration of pregnancy was significantly prolonged compared with group A and C, and occurrence of pPROM was significantly lower. No significant differences were found in those rates between groups A and C. CONCLUSION: Successful treatment for women with IL-8 elevations in cervical mucus decreased rates of premature birth or pPROM.     

灭滴灵阴道用药对细菌性阴道病晚孕妇女不良妊娠结局的预防效果

目的：探讨灭滴灵治疗细菌性阴道病对不良妊娠结局是否有改善作用。方法：在产科门诊对用阴道分泌物涂片革兰氏染色Ｎｕｇｅｎｔ诊断标准孕２８－３２周妇女进行细菌性阴道病筛查，对检出的患者随机分为二组；观察组５２例，未给予治疗；治疗组５０例，采用灭滴灵阴道用药治疗，必要时再次治疗。     

Bacterial vaginosis. Transmission, role in genital tract infection and pregnancy outcome: an enigma

Whether bacterial vaginosis (BV) is acquired from an endogenous or an exogenous source is subject to controversy. Despite findings of an association between sexual behaviour and BV, some data indicate that BV is not a sexually transmitted infection in the traditional sense, while other data indicate that BV is an exogenous infection. A third aspect of BV is its tendency to go unnoticed by affected women. All of this will have a strong impact on how physicians view the risks of asymptomatic BV. This review focuses on whether or not BV should be regarded as a sexually transmitted infection (STI), its role in postoperative infections and pelvic inflammatory disease (PID), and on whether or not treatment of BV during pregnancy to reduce preterm delivery should be recommended. The reviewed studies do not lend unequivocal support to an endogenous or exogenous transmission of the bacteria present in BV. For women undergoing gynaecological surgery such as therapeutic abortion, the relative risk of postoperative infection is clearly elevated (approx. 2.3-2.8). A weaker association exists between BV and pelvic inflammatory disease. Data on treatment of BV as a way of reducing preterm delivery are inconclusive and do not support recommendations for general treatment of BV during pregnancy. The discrepant associations between BV and preterm birth found in recent studies may be explained by variations in immunological response to BV. Genetic polymorphism in the cytokine response--both regarding the TNF alleles and in interleukin production--could make women more or less susceptible to BV, causing different risks of preterm birth. Thus, studies on the vaginal inflammatory response to microbial colonization should be given priority.     

Trimethylamine content in vaginal secretion and its relation to bacterial vaginosis

The presence of a fishy odor emanating from women who present with a malodorous vaginal discharge is well known. The odor is due to bacterial reduction of trimethylamine oxide to trimethylamine (TMA) in vaginal secretion. The release of TMA from specimens of vaginal fluid following the addition of alkali is often used in making a clinical diagnosis of bacterial vaginosis (BV). We now report a sensitive gas chromatographic method for analysis and quantification of TMA in vaginal fluid in which weighed samples were used. In addition, a proper diagnosis of BV was obtained using Gram-stained smears of the vaginal fluid according to the method of Nugent et al. (R. P. Nugent et al., J Clin Microbiol 1991;29:297-301). We also diagnosed BV according to Hallén et al. (A. Hallén et al. Genitourin Med 1987;63:386-9). TMA was present in all women with a Nugent score between 7 and 10 and in almost all women diagnosed with BV according to the method of Hallén et al. TMA was not found or was only found in very low concentrations in vaginal fluid from women with Nugent scores of 0 to 3. TMA was also found in four women with a negative sniff test. It seems that high levels of TMA in samples of vaginal fluid are typical for BV regardless of the scoring method used for diagnosis. However, low levels of TMA, <5 microg/g vaginal fluid, do not always correlate with BV.     

Interleukin-8 levels and granulocyte counts in cervical mucus during pregnancy

PROBLEM: We measured interleukin-8 (IL-8) and granulocyte counts in cervical mucus to assess local immunity and parturition. METHOD OF STUDY: We detected IL-8 by enzyme linked immunosorbent assay (ELISA) and Western blotting. Granulocytes were counted on a slide glass containing mucus from the external cervical os. RESULTS: ELISA and Western blotting revealed IL-8 in cervical mucus from both nonpregnant and pregnant women. There were no significant differences in cervical mucus IL-8 levels or granulocyte counts between follicular, ovulatory, and luteal phases. However, IL-8 and granulocyte counts were significantly decreased after menopause. IL-8 and granulocyte counts were increased significantly during pregnancy, and were further increased after 38 weeks of gestation and at labor. IL-8 levels were significantly correlated with granulocyte counts, based on the study of 678 samples (P <0.0001). CONCLUSION: IL-8 is involved in the increase of cervical granulocytes and in the process of parturition.     

Reciprocal Interference between Lactobacillus spp. and Gardnerella vaginalis on Initial Adherence to Epithelial Cells

Bacterial vaginosis (BV) is the most common vaginal disorder in women of child-bearing age. It is widely accepted that the microbial switch from normal microflora to the flora commonly associated with BV is characterized by a decrease in vaginal colonization by specific Lactobacillus species together with an increase of G. vaginalis and other anaerobes. However, the order of events leading to the development of BV remains poorly characterized and it is unclear whether the decrease in lactobacilli is a cause or a consequence of the increase in the population density of anaerobes. Our goal was to characterize the interaction between two Gardnerella vaginalis strains, one of which was isolated from a healthy woman (strain 5-1) and the other from a woman diagnosed with BV (strain 101), and vaginal lactobacilli on the adherence to cervical epithelial cells. In order to simulate the transition from vaginal health to BV, the lactobacilli were cultured with the epithelial cells first, and then the G. vaginalis strain was introduced. We quantified the inhibition of G. vaginalis adherence by the lactobacilli and displacement of adherent lactobacilli by G. vaginalis. Our results confirmed that pathogenic G vaginalis 101 had a higher capacity for adhesion to the cervical epithelial cells than strain 5-1. Interestingly, strain 101 displaced L. crispatus but not L. iners whereas strain 5-1 had less of an effect and did not affect the two species differently. Furthermore, L. iners actually enhanced adhesion of strain 101 but not of strain 5-1. These results suggest that BV-causing G. vaginalis and L. iners do not interfere with one another, which may help to explain previous reports that women who are colonized with L. iners are more likely to develop BV.     

Interrelationships of interleukin-8 with interleukin-1beta and neutrophils in vaginal fluid of healthy and bacterial vaginosis positive women

Vaginal innate immunity in response to microbial perturbation is still poorly understood and could be crucial for protection from adverse outcomes. We investigated the relationship between interleukin (IL)-8, IL-1beta and neutrophils in vaginal fluid obtained from 60 healthy women and 51 women who were bacterial vaginosis (BV) positive. Concentrations of IL-8 and IL-1beta were highly correlated with counts of neutrophils in vaginal fluid of the entire population examined (111 subjects). Vaginal IL-1beta concentrations were significantly higher (P < 0.001) in BV positive women. There was no significant difference in IL-8 levels or number of neutrophils between healthy controls and BV positive women. None of the healthy controls with high neutrophil counts (> or =75th percentile, 14 average count per field) had high concentrations of IL-1beta (> or =75th percentile, 220 pg/ml), whereas 84% of BV positive women with high neutrophil counts had high IL-1beta concentrations (P < 0.001). On the contrary, no difference in the percentage of subjects with elevated concentrations of IL-8 (> or =75th percentile, 2842 pg/ml) was found between healthy and BV positive women with high numbers of neutrophils (55.5% of healthy versus 53% of BV positive women). Our findings show that BV causes a large increase in IL-1beta concentrations which is not paralleled by an increase in IL-8 concentrations in vaginal fluid, suggesting that BV-associated factors more specifically dampen IL-8 rather than IL-1beta. The lack of an increase in IL-8 may explain the absence of an increase in neutrophil numbers in most women exposed to abnormal vaginal colonization (BV).     

羊水中白细胞介素-8水平与绒毛膜羊膜炎的关系

目的探讨胎膜早破孕妇羊水中白细胞介素-8（IL-8）水平与绒毛膜羊膜炎的关系。方法用酶联免疫法（ELISA法）测定62例胎膜早破孕妇和46例正常足月妊娠未临产孕妇羊水中IL-8浓度;病理检查两组分娩后的胎膜组织,确定有无绒毛膜羊膜炎。结果胎膜早破组羊水中IL-8浓度高于对照组（P〈0.05）,破膜时间超过24h其羊水中IL-8水平明显高于破膜时间小于12h内的患者,绒毛膜羊膜炎组羊水中IL-8浓度高于对照组（P〈0.05）。结论胎膜早破及绒毛膜羊膜炎孕妇羊水中IL-8水平显著升高,可作为绒毛膜羊膜炎的早期诊断指标。     

Normal flora and bacterial vaginosis in pregnancy: an overview

Abstract

The female genital tract is an intricate, yet balanced ecosystem that hosts a variety of different residential microflora. The physiological changes that occur during pregnancy may disrupt this balanced ecosystem and predispose women to a potentially pathogenic microbiota. Bacteria that are associated with bacterial vaginosis (BV) are opportunistic pathogens that frequently form part of this microbiota. The overgrowth of and infections with these bacteria are linked to poor obstetric outcomes and increased transmission of other reproductive tract infections (RTIs). These infections increase women's susceptibility of acquiring HIV, the rates of HIV shedding and the development of acquired immune deficiency syndrome (AIDS) in HIV-infected patients. It is unknown how the plethora of bacterial species associated with BV contributes to the dynamics of this condition. The use of high-throughput methods have led to the in-depth investigation of different BV-related bacterial species and the functional capabilities of these species. However, the pathogenesis of BV is still poorly defined and the role of individual BV-related bacterial species in specific pregnancy complications is unclear and controversial. The majority of BV infections are asymptomatic and successful diagnosis is complicated by the lack of reliable and standardized diagnostic tests.     

REVIEW ARTICLE: Th1/Th2/Th17 and Regulatory T‐Cell Paradigm in Pregnancy

Citation Saito S, Nakashima A, Shima T, Ito M. Th1/Th2/Th17 and regulatory T‐cell paradigm in pregnancy. Am J Reprod Immunol 2010 T‐helper (Th) cells play a central role in modulating immune responses. The Th1/Th2 paradigm has now developed into the new Th1/Th2/Th17 paradigm. In addition to effector cells, Th cells are regulated by regulatory T (Treg) cells. Their capacity to produce cytokines is suppressed by immunoregulatory cytokines such as transforming growth factor (TGF)‐β and interleukin (IL)‐10 or by cell‐to‐cell interaction. Here, we will review the immunological environment in normal pregnancy and complicated pregnancy, such as implantation failure, abortion, preterm labor, and preeclampsia from the viewpoint of the new Th1/Th2/Th17 and Treg paradigms.     

Uterine cervical inflammatory cytokines, interleukin-6 and -8, as predictors of miscarriage in recurrent cases

PROBLEM: Inflammatory changes frequently occur in cases of second trimester miscarriage or pre-term delivery, but little attention has been paid to this association with recurrent miscarriage. As interleukin-6 and interleukin-8 are inflammatory cytokines reported to be associated with bacterial vaginosis, intrauterine infections, and pre-term delivery, we here investigated whether they might have predictive value for spontaneous abortion in recurrent cases. METHOD OF STUDY: Cervical mucus and sera were collected at 4-5 weeks' gestation from a total of 59 patients with a history of two or more unexplained consecutive first trimester miscarriages, and examined by enzyme-linked immunosorbent assay. Patients then were followed up without medication and their pregnancy outcomes were compared with the test results. RESULTS: Of a total of 59 patients, 13 (22%) miscarried subsequently. Both IL-6 and IL-8 in cervical mucus were significantly higher in patients who miscarried subsequently than in those who had a live birth. In addition, there was no correlation between cervical mucus and serum concentrations of IL-6 and IL-8 take at the same time, and there was no relation with serum IL-6 and IL-8 levels between the two groups. CONCLUSION: Cervical IL-6 and IL-8 might have predictive value for cases of recurrent miscarriage.     

Identification of immune cells by flow cytometry in vaginal lavages from women with vulvovaginitis and normal microflora

Citation Giraldo PC, de Carvalho JBJ, do Amaral RLG, da Silveira Gonçalves AK, Eleutério J Jr, Guimarães F. Identification of Immune Cells by Flow Cytometry in Vaginal Lavages from Women with Vulvovaginitis and Normal Microflora. Am J Reprod Immunol 2012; 67: 198–205 Problem The extent of the vaginal immune response is not fully determined. The purpose of this study was to evaluate the vaginal immune cells from women with vulvovaginitis (VV). Method of Study A total of 142 volunteers diagnosed with bacterial vaginosis (BV), vulvovaginal candidiasis (VC), and BV associated with VC or normal microflora were sampled to evaluate the immune cells by flow cytometry. The immune cells were obtained by vaginal lavage and labeled with fluorochrome‐conjugated monoclonal antibodies to identify neutrophil granulocytes, macrophages, CD4⁺ and CD8⁺ T lymphocytes, B lymphocytes, and NK lymphocytes. Results Neutrophil granulocytes were present in 84.6% of samples among the leukocyte populations. Considering samples in which neutrophils were present, the mean percentage of neutrophil granulocytes was significantly higher in women with VC than BV and normal microflora and was significantly lower in women with BV than normal microflora. Macrophages and lymphocytes were present in a lower percentage of samples. The mean percentage of CD4⁺ T lymphocytes in vaginal lavages was significantly higher in VC and BV compared with women with normal microflora. Conclusions Neutrophils were the predominant leukocytes and were associated with VC and inversely with BV. CD4⁺ T lymphocytes were associated with both VC and BV.     

胃黏膜线粒体DNA不稳定与白细胞介素8活性的关系

目的　评价胃黏膜线粒体DNA不稳定 (mtMSI)与组织白细胞介素 8(IL 8)活性的关系。方法　采用聚合酶链反应 单链构象多态性分析 (PCR SSCP)方法检测mtMSI;组织IL 8含量测定采用ELISA方法。结果　 3 0例胃癌检出mtMSI 11例 ( 3 6 7% ) ,15例肠化胃黏膜组织中有 2例为mtMSI,10例异型增生黏膜组织有 2例为mtMSI,10例萎缩性胃炎黏膜组织有 1例检出mtMSI。胃黏膜细胞IL 8水平在mtMSI阳性组 [( 76 8± 3 8)pg/mg]显著高于mtMSI阴性组 [( 48 3± 3 6)pg/mg ,P <0 0 5 ]。结论　胃黏膜细胞mtMSI可能与IL 8释放有关 ,并参与了胃癌的发生     

Neuropeptides and interleukin-6 in human joint inflammation. Relationship between intraarticular substance P and interleukin-6 concentrations

Plasma and synovial fluid concentrations of interleukin-6 (IL-6), using an enzyme-linked immunosorbent assay, as well as immunoreactive levels of calcitonin gene-related peptide (CGRP), substance P and vasoactive intestinal peptide (VIP) were measured in 18 patients with rheumatoid arthritis and 20 with osteoarthritis of the knee. The concentrations of IL-6 were elevated in both plasma and synovial fluids from patients with rheumatoid arthritis whereas higher levels of substance P-, CGRP- and VIP-like immunoreactivities were found in the synovial fluid, but not in plasma, from patients with rheumatoid arthritis when compared with those in osteoarthritis. Furthermore, IL-6 and substance P levels in synovial fluid were significantly correlated both in rheumatoid arthritis and osteoarthritis patients. Our data seem to support the idea of an important role shared by neuropeptides and IL-6 in the pathogenesis of human inflammatory joint disease.     

Gardnerella vaginalis and Lactobacillus sp in liquid-based cervical samples in healthy and disturbed vaginal flora using cultivation-independent methods

Our objective was to determine the morphotype of the adherent bacteria in liquid-based cytology (LBC) in smears with healthy and disturbed vaginal flora. And to use PCR technology on the same fixed cell sample to establish DNA patterns of the 16S RNA genes of the bacteria in the sample. Thirty samples were randomly selected from a large group of cervical cell samples suspended in a commercial coagulant fixative "(BoonFix)." PCR was used to amplify DNA of five bacterial species: Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus jensenii, Gardnerella vaginalis, and Mycoplasma hominis. The LBC slides were then analyzed by light microscopy to estimate bacterial adhesion. DNA of lactobacilli was detected in all cell samples. Seventeen smears showed colonization with Gardnerella vaginalis (range 2.6 x 10(2)-3.0 x 10(5) bacteria/mul BoonFix sample). Two cases were identified as dysbacteriotic with high DNA values for Gardnerella vaginalis and low values for Lactobacillus crispatus. The sample with the highest concentration for Gardnerella vaginalis showed an unequivocal Gardnerella infection. This study indicates that the adherence pattern of a disturbed flora in liquid-based cervical samples can be identified unequivocally, and that these samples are suitable for quantitative PCR analysis. This cultivation independent method reveals a strong inverse relationship between Gardnerella vaginalis and Lactobacillus crispatus in dysbacteriosis and unequivocal Gardnerella infection.