Gebelik süresince uterus ve lenfoid organlardaki dendritik hücrelerin karakterizasyonu

Amaç: Bu çalışmanın amacı gebeliğin 15`inci gününde dendritik hücrelerin fenotiplemesi, sitokin ve kemokin salınımın araştırılmasıdır.Gereç ve Yöntem: Desidual hücreler izole etmek için, Uterus plasenta ve fetüsten ayrılarak diseke edildi. Uterustan elde edilen tek hücreli süspansiyonlar, CD45, CD11c ve MHC sınıf II gibi DH yüzey moleküllerine karşı antikorlarla inkübe edildi. Flow sitometri ile hücreler ayıklandı. Konfokal lazer scanning mikroskop ile incelendi. Desidua, dalak ve uterus lenf düğümlerindeki dendritik hücreler izole edilerek ayni şekilde boyandı ve flow sitometri ile analiz edildi. Dendritik hücrelerin PCR analizi yapıldı.Bulgular ve Sonuçlar: Gebelik sırasında uterustaki, lenf nodlarındaki ve dalaktaki dendritik hücreler arasında farklılıklar gözlendi. Gebe farelerin uterus ve periferik lenf dokularındaki CD45+ CD11c+ hücrelerinin MHC sınıf II’nin ve yardımcı moleküllerin ekspresyonunun gebe olmayanlardan farklı olmadığı gösterildi. CD11b- CD8+ olan uterustaki ve uterus lenf düğümlerindeki CD11c+ hücrelerinin oranında gebeliğe bağlı bir azalma görülmedi. Hamilelikte, CD11b+ alt kümelerini ve uterus DH’lerinde kemokin ligand 6'nın ekspresyonunu arttırdığı gözlendi. DH alt kümeleri lenfatik hücrelerin homeostazında önemli olan sitokin interlökin-15 açısından dokuya ve gebeliğe bağlı olarak değişken ekspresyonlar gösterdiği tespit edildi.Objective: This study aims to investigate the dendritic cells phenotype, cytokine and chemokine release on the 15th day of pregnancy.Materials and Methods: The decidual cells were isolated, the samples were separated from the placenta and fetüs. Singel cell suspensions obtained from the uterus were incubated with antibodies to DC surface molecules such as CD45, CD11c and MHC sınıf II. The cells isolated via flow cytometry sorting. Confocal laser scanning microscope was used to study dendritic cells. Decidual cell, spleen and uterine draining lymph node isolated, stained and analyzed by flow cytometry. RT-PCR analysis of dendritic cells was performed.Results and Conclusion: Differences among DCs in the uterus, uterine draining nodes and spleen, were observed. CD45+ CD11c+ cells in the uterus and peripheral lymphoid tissue of pregnant mice, expression of major histocompatibility complex class II (MHC sınıf II) and costimulatory molecules was similar to that in the non-pregnant state. There was no pregnancy-related decrease in the proportion of CD11c+ cells in the uterus or in the uterine node that were CD11b- CD8+. Pregnancy increased the CD11b+ subsets and the expression of chemokine ligand 6 in DCs of the uterine draining nodes. DC subsets showed variable expression, with respect to tissue and pregnancy, of the cytokine interleukin-15, which is important in lymphoid cell homeostasis

Pregnancy Alters the Proliferation and Apoptosis of Mouse Splenic Erythroid Lineage Cells and Leukocytes1

Pregnancy induces dynamic changes in the maternal environment that include reversible modifications in response to systemic mediators and local signals. The spleen can be used to determine the effects of pregnancy on multiple cellular populations, including those of the erythroid lineage and the immune system. Current evidence suggests that the transient increase in the size of the spleen during pregnancy is due to the expansion of erythroid precursors. However, it is unclear what factors contribute to this increase. Moreover, the additional erythroid cells may compete with neighboring leukocytes for growth factors or space, and this may in turn alter the function of these populations. Therefore, we assessed proliferation and apoptosis throughout gestation using in vivo bromodeoxyuridine incorporation and the TUNEL assay, respectively. Here, we show that erythroid-lineage TER-119+ cells expanded significantly in midgestation because of enhanced proliferation and diminished apoptosis. This correlated with increased expression of the erythropoietin receptor (Epor) and decreased expression of the death receptor Fas, respectively. Leukocytes demonstrated population-specific responses. Natural killer cells proliferated in early pregnancy. Both lymphocytes and CD11B+ cells underwent enhanced proliferation during midgestation. In contrast, neutrophils exhibited augmented proliferation throughout pregnancy. These subset-specific alterations in proliferation and death in the spleen suggest that complex regulation of population dynamics exists during pregnancy

A homozygous splice variant in ATP5PO , disrupts mitochondrial complex V function and causes Leigh syndrome in two unrelated families

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High Rate of Recurrent De Novo Mutations in Developmental and Epileptic Encephalopathies

Developmental and epileptic encephalopathy (DEE) is a group of conditions characterized by the co-occurrence of epilepsy and intellectual disability (ID), typically with developmental plateauing or regression associated with frequent epileptiform activity. The cause of DEE remains unknown in the majority of cases. We performed whole-genome sequencing (WGS) in 197 individuals with unexplained DEE and pharmaco-resistant seizures and in their unaffected parents. We focused our attention on de novo mutations (DNMs) and identified candidate genes containing such variants. We sought to identify additional subjects with DNMs in these genes by performing targeted sequencing in another series of individuals with DEE and by mining various sequencing datasets. We also performed meta-analyses to document enrichment of DNMs in candidate genes by leveraging our WGS dataset with those of several DEE and ID series. By combining these strategies, we were able to provide a causal link between DEE and the following genes: NTRK2, GABRB2, CLTC, DHDDS, NUS1, RAB11A, GABBR2, and SNAP25. Overall, we established a molecular diagnosis in 63/197 (32%) individuals in our WGS series. The main cause of DEE in these individuals was de novo point mutations (53/63 solved cases), followed by inherited mutations (6/63 solved cases) and de novo CNVs (4/63 solved cases). De novo missense variants explained a larger proportion of individuals in our series than in other series that were primarily ascertained because of ID. Moreover, these DNMs were more frequently recurrent than those identified in ID series. These observations indicate that the genetic landscape of DEE might be different from that of ID without epilepsy