Data for: Latent HCMV is Associated with Specific Proinflammatory Immune Phenotype in Women with Implantation Failure
Description
Latent HCMV infection specifically influences immune system and results in pro-inflammatory phenotype formation in part of infected population. It was found that accentuations in immune phenotype of HCMV-seropositive women are very similar to previously described in association with reproductive failures but without HCMV-serostatus taken into account. Study population Infertile women (n=470) who were eligible for IVF/ET were selected for this study. Patients included in the study had at least 1 case of idiopathic IVF failure (average 2.32) or at least 1 case of idiopathic first trimester pregnancy failure (average 2.04). Patients had no supplemental treatment with immunoglobulins or corticosteroids during last 3 months. None of the patients had active autoimmune disease or history of autoimmune diseases. None of the patients had active infection within 6 previous months. Methods ELISA Anti-HCMV IgG and IgG avidity were measured in patient’s serum by VitoTest ELISA system (Ukraine) according to instructions. Presence of low avidity IgG of more than 50% was considered as active HCMV infection. 2.3 Flow Cytometry of Lymphocyte Subsets Lymphocyte subsets were identified by three-color flow cytometry using erythrocyte-lysing whole blood method of lymphocyte staining by FITC-, PE- or PE-Cy5-conjugated monoclonal antibodies (BD Bioscience, San Jose, USA). Stained samples were lysed, washed and analyzed by FACScan flow cytometer using CellQuest software (BD Bioscience, San Jose, USA). 2.4 NK cytotoxicity assays Flow cytometry NK cytotoxicity assay (FCA) was described previously [15]. We used Calcein-AM (Sigma) labeling K562 as target cells. Dead cells after 4h incubation were labeled by propidium iodide.