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Although there are some reports about infected mother’s immunity during pregnancy, it’s role in promoting or inhibiting congenital transmission has not been directly tested (6C10)

Although there are some reports about infected mother’s immunity during pregnancy, it’s role in promoting or inhibiting congenital transmission has not been directly tested (6C10). Other important players involved in the control of during acquired infections, are human IgG subclasses -predominantly IgG1- and their Fc receptors; importantly, specific IgG1 in infected mothers has been related to clinical problems in their congenitally infected babies; however, they were measured at delivery, when transmission already occurred (11). Due to the lack of information about the specific lymphocyte populations, cytokines, and antibody subtypes induced by in infected women during pregnancy and their relationship to vertical transmission, we performed the present study. Materials and Methods Ethical Aspects This work was carried out in accordance with the World Medical Association’s Declaration of Helsinki. approximately one-third of all humans (1). Most individuals with toxoplasmosis show no clinical signs, however immunodeficient and congenitally infected patients may develop pathological conditions (2). Congenital contamination occurs due to vertical transmission of during pregnancy, and although it is usually asymptomatic and self-limited in the mother, if the fetus is usually infected, he/she may develop variable clinical features, such Salicin (Salicoside, Salicine) as spontaneous abortion, stillbirth, hydrocephalus, macro or microcephalus, cerebral calcifications, retinochoroiditis, and other ocular or central nervous system alterations, which can manifest even years later in life (3). It is widely known that in immunocompetent individuals, such as pregnant women, a Th1-type immune response represents the main effective response against the parasite (4, 5). Although there are some reports about infected mother’s immunity during pregnancy, it’s role in promoting or inhibiting congenital transmission has not been directly tested (6C10). Other important players involved in the control of during acquired infections, are human IgG subclasses -predominantly IgG1- and their Fc receptors; importantly, specific IgG1 in infected mothers has been related to clinical problems in their congenitally infected babies; however, they were measured at delivery, when transmission already occurred (11). Due to the lack of information about the specific lymphocyte populations, cytokines, and antibody subtypes induced by in infected women during pregnancy and their relationship to vertical transmission, we performed the present study. Materials and Methods Ethical Aspects This work was carried out in accordance with the World Medical Association’s Declaration of Helsinki. It contains partial results from the project 060/2011, approved by the Research and Research Ethics Boards of the Instituto Nacional de Pediatra (INP), Mexico City, Mexico; registered at the Office for Human Research Protection of the NIH (http://ohrp.cit.nih.gov/search/search.aspx) with numbers IRB00008064 and IRB00008065. It was also approved by the INP Committee of Laboratory Animal Use and Care; approval is available upon request. The Instituto Nacional de Perinatologa (INPer) also approved the project (number 212250C02231). All participants signed an informed consent, which explicitly stated that it was of low risk, considering that clinical management was not altered for the protocol. All newborns were clinically managed at INP according to national and international standards. Biosafety steps were carefully followed, in order to avoid technician’s contamination with the strain used to prepare the antigen, by using a level II biohazard hood (Labconco Purifier Class II Biosafety Cabinet, Labconco Corp., Kansas, MO) when working with the parasites. A well-controlled animal house is present at INP, where the mice are inoculated. In Salicin (Salicoside, Salicine) addition, to avoid, potentially contagious diseases (present in the women recruited), only trained personnel who wore gloves and face masks was Rtn4r authorized to take samples from patients. Parasite Antigen tachyzoites (RH strain) were maintained in BALB/c mice by intraperitoneal passages. Peritoneal exudates from 40 mice were harvested and washed twice (720 g, 10 min, 4C) in PBS supplemented with a protease cocktail inhibitor (10 mg/ml aprotinin, 50 g/ml leupeptin, and 1.6 mmol/L phenylmethylsulfonyl fluoride). To prepare soluble antigen (STAg), the parasite suspension was lysed by five sonication cycles (60 Hz for 1 min each) on ice. After centrifugation (10,000 g, 2 h, 4C) supernatants were collected and sterilized by filtration through a 0.2 m-pore size membrane (Corning Costar Corp., Cambridge, MA). The protein concentration was determined by Bradford (Quick Start? Protein Assay, Bio-Rad laboratories, Hercules, Salicin (Salicoside, Salicine) CA) and aliquots were stored at ?80C until use. Patients and Study Strategy From 1,083 pregnant women screened for toxoplasmosis, we recruited 11 of them who agreed to participate and met criteria for further analysis. They were patients of the Instituto Nacional de Perinatologa-Isidro Espinosa de los Reyes (third level hospital) or the Centro de Salud-Dr. Gustavo.