Evaluation of immunoregulatory and apoptosis and gene polymorphisms in patients with preeclampsia.

Purpose: to evaluate the relationship between CTLA-4 (+49), CD28 (+17), ICOS (-1564), Fas (-670) and FasL (-844) gene polymorphisms and susceptibility to preeclampsia (PE) in its mild and severe form. Methods: This case-control study included 130 preeclamptic women and 261 control pregnant women without any obstetric or systemic disorders, with a history of at least two previous successful pregnancies. Genomic DNA was extracted from whole blood using DTAB/CTAB method, and polymorphisms genotyping were obtained by PCR-RFLP technique. Data were analyzed by Student´s t, qui-square (c2) and Fischer´s exact tests, and the level of significance was set at p<0.05. Results: Genotype frequencies of ICOS (-1564) polymorphism were 41.9% TT, 48.4% TC and 9.7% CC in the PE group; and 59.3% TT, 32.3% TC and 8.4% CC in controls (p=0.01). Genotype frequencies of Fas (-670) polymorphism were 19.8% AA, 45.2% AG and 34.9% GG in the PE group; and 62% AA, 50.2% AG and 24.5% GG in controls (p=0.09). FasL (-844) genotype frequencies were 23.8% TT, 42.3% TC and 33.8% CC in the PE group; and 33.5% TT, 46.5% TC and 20% CC in controls (p=0.007). There were no significant differences between CTLA-4 and CD28 genotypic frequencies, when comparing women with PE and controls (c2 test: p=0.62, p=0.38, respectively). When comparing mild and severe PE, there were significant differences in CTLA-4 polymorphism (c2 test: p=0.03). There were no significant differences detected between the clinical patterns of PE in CD28, ICOS, Fas and FasL polymorphisms genotypic distribution (c2 test: p=0.33, p=0.28, p=0.5 and p=0.91 respectively). Conclusions: There is evidence of an association between ICOS (-1564), Fas (- 670) and Fas-L (-844) polymorphisms and the occurrence of PE. Our data suggests that CTLA- 4 (+49) polymorphism is related to the occurrence of different clinical patterns of PE, i.e., ifmild or severe forms. We did not identify any association between CD28 (+17) polymorphism and PE.Objetivo: avaliar a relação entre os polimorfismos de genes que codificam CTLA-4 (+49), CD28 (+17), ICOS (-1564), Fas (-670) e FasL (-844) e a suscetibilidade à pré-eclâmpsia (PE) em suas formas clínicas: leve e grave. Pacientes e métodos: Este estudo teve caráter tipo caso-controle e incluiu 130 pacientes com PE e 261 gestantes sem patologia sistêmica ou obstétrica, e com história de duas ou mais gestações sem intercorrências como controles. O DNA genômico foi extraído de sangue periférico por método de DTAB/CTAB, e as genotipagens foram realizadas por técnica de PCR-RFLP. Foram utilizados os testes t de Student, qui-quadrado (c2) e exato de Fischer para a análise dos resultados, tendo sido adotado o nível de significância de p<0,05. Resultados: As frequências genotípicas de ICOS (- 1564) foram 41,9% TT, 48,4% TC e 9,7% CC em casos de PE; e 59,3% TT, 32,3% TC e 8,4% CC em controles (p=0,01). As frequências genotípicas de Fas (-670) foram 19,8% AA, 45,2% AG e 34,9% GG em pacientes com PE; e 62% AA, 50,2% AG e 24,5% GG no grupo controle (p=0,09). As frequências genotípicas de FasL (-844) foram 23,8% TT, 42,3% TC e 33,8% CC em pacientes com PE; e 33,5% TT, 46,5% TC e 20% CC no grupo controle (p=0,007). Não foram identificadas diferenças significantes entre as frequências genotípicas de CTLA-4 e CD28 quando comparados os grupos de PE e controle entre si (Teste de c2: p=0,62, p=0,38, respectivamente). Na comparação entre PE leve e grave observamos diferença significante relacionada ao polimorfismo de CTLA-4 (Teste de c2: p=0,03). Não identificamos diferença entre as formas clínicas quanto a distribuição genotípica dos polimorfismos de CD28, ICOS, Fas e FasL (Teste de c2: p=0,33, p=0,28, p=0,5 e p=0,91 respectivamente). Conclusão: Foi observada associação entre os polimorfismos dos genes codificadores de ICOS (-1564), Fas (-670) e Fas-L (-844) e a ocorrência de pré-eclâmpsia. Nossos dados sugerem que o polimorfismo de CTLA-4 (+49) está relacionado à ocorrência de diferentes formas clínicas da doença, se grave ou leve. Não identificamos qualquer associação entre a PE e o polimorfismo de CD28 (+17).TEDEBV UNIFESP: Teses e dissertaçõe

Cervical cerclage placement decreases local levels of proinflammatory cytokines in patients with cervical insufficiency

BACKGROUND: Cervical insufficiency is characterized by premature, progressive dilation and shortening of the cervix during pregnancy. If left unattended, this can lead to the prolapse and rupture of the amniotic membrane, which usually results in midtrimester pregnancy loss or preterm birth. Previous studies have shown that proinflammatory cytokines such as interleukin-1 beta, interleukin-6, interleukin-8, and tumor necrosis factor alpha are up-regulated in normal parturition but are also associated with preterm birth. Studies evaluating such markers in patients with cervical insufficiency have evaluated only their diagnostic potential. Even fewer studies have studied them within the context of cerclage surgery. OBJECTIVES(S): The objective of the study was to evaluate the impact of local and systemic inflammatory markers on the pathogenesis of cervical insufficiency and the effect of cerclage surgery on the local immune microenvironment of women with cervical insufficiency. STUDY DESIGN: We recruited 28 pregnant women (12- 20 weeks' gestation) diagnosed with insufficiency and referred for cerclage surgery and 19 gestational age-matched normal pregnant women as controls. Serum and cervicovaginal fluid samples were collected before and after cerclage surgery and during a routine checkup for normal women and analyzed using a targeted 13-plex proinflammatory cytokine assay. RESULTS: Before surgery, patients with cervical insufficiency had higher levels of interleukin-1b, interleukin-6, interleukin-12, monocyte chemoattractant protein-1 and tumor necrosis factor alpha in cervicovaginal fluid compared to controls, but after surgery, these differences disappeared. No differences were found in serum of insufficiency versus control women. In patients with insufficiency, the levels of interleukin-1b, interleukin-6, interleukin-8, monocyte chemoattractant protein-1, and interferon gamma in cervicovaginal fluid declined significantly after cerclage compared with before intervention, but these changes were not detected in serum. CONCLUSION: Compared with normal women, patients with cervical insufficiency have elevated levels of proinflammatory cytokines in cervicovaginal fluid but not in serum, suggesting a dysregulation of the local immune environment. Cerclage intervention led to a significant decline in these proinflammatory cytokines, suggesting that cerclage may help reduce local inflammation in cervical insufficiency.Canadian Institutes of Health ResearchMITACS GlobalinkQueens Univ, Dept Biomed & Mol Sci, Kingston, ON, CanadaUniv Fed Sao Paulo, Escola Paulista Med, Dept Obstet, Sao Paulo, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Dept Obstet, Sao Paulo, BrazilWeb of Scienc

FAS and FAS-L Genotype and Expression in Patients With Recurrent Pregnancy Loss

We assessed FAS and FAS-L gene polymorphisms and messenger RNA (mRNA) levels in patients with recurrent pregnancy loss (RPL). This case-control study compared 129 women with RPL with 235 healthy multiparous women (control group). Genomic DNA and total mRNA were extracted from whole blood, and polymorphisms genotyping was performed by polymerase chain reaction (PCR). Messenger RNA expression levels were analyzed by real-time PCR. Data were analyzed by chi-square and Fisher exact tests; P < .05 was considered significant. There were no significant differences in the FAS (670 A/G) genotype or allelic frequencies between the RPL and control groups. We found significant differences in the FAS-L (844 C/T) genotype and allelic frequencies between women with RPL and controls. Patients with RPL had significantly higher FAS-L expression. Our data suggest that FAS-L gene polymorphism is associated with increased susceptibility to RPL. Moreover, women with RPL seem to abnormally express FAS-FAS-L molecules.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Obstet, BR-04037001 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Obstet, BR-04037001 São Paulo, BrazilFAPESP: 09/50236-5CNPq: 301886/2009-1Web of Scienc

Association Study of Vascular Endothelial Growth Factor and Polymorphisms of its Gene with Ectopic Pregnancy

ProblemIn ectopic pregnancy, increased levels of vascular endothelial growth factor are present. the aims of this study were to determine the association between -634C/G, -460T/C, and +936C/T vascular endothelial growth factor (VEGF) polymorphisms and ectopic pregnancy, and to determine whether serum levels of VEGF were affected by genetic factors.Method of studyThis is a case-control study wherein 74 women with a history of ectopic pregnancy in a tertiary care center were compared to 134 post-menopausal controls with two pregnancies and no ectopic pregnancy for the genotyping of VEGF polymorphisms. for 35 patients with the diagnosis of ectopic pregnancy, serum concentrations of VEGF were obtained before the treatment. Genotyping of VEGF (-634C/G, -460T/C, and +936C/T) polymorphisms was performed by PCR, followed by endonuclease digestion. ELISA was performed to evaluate the VEGF serum levels.ResultsThe -634C/G, -460T/C, and +936C/T VEGF polymorphisms were not associated with ectopic pregnancy (P = 0.170, P = 0.285, and P = 0.700, respectively). the serum levels of VEGF were not associated with the genotype of -634C/G, -460T/C, and +936C/T VEGF polymorphisms (P = 0.702; P = 0.347, and P = 0.256, respectively).ConclusionThere was no association between ectopic pregnancy and -634C/G, -460T/C, and +936C/T VEGF polymorphisms. There was no correlation between VEGF genotype and the expression of VEGF in blood samples.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Obstet, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Obstet, São Paulo, BrazilWeb of Scienc