S.), and the National Institutes JNJ-7706621 of Health (Loan Repayment Program for Pediatric Research to F. and infants, irrespective of perinatal HIV exposure or infant CMV status at 6 months of age. However, the glycoprotein BCspecific IgG titer, measured by 2 unique assays, was higher in infants without CMV contamination and was moderately associated with delayed CMV acquisition. Conclusions These data suggest that high levels of glycoprotein BCspecific IgG may contribute to the partial protection against postnatal CMV contamination afforded by maternal antibodies, and they support the continued inclusion of glycoprotein B antigens in CMV vaccine candidates. Keywords: Cytomegalovirus, Mouse monoclonal to WDR5 immune correlates, vaccine Congenital cytomegalovirus (CMV) contamination occurs in 1 in 150 live births, and approximately 15% of infected infants develop permanent neurologic sequelae [1, 2]. Owing to the high disease burden of congenital CMV, in 2000 the National Academy of Medicine declared development of a CMV vaccine a tier 1 priority [3]. However, a poor understanding of immune correlates that protect against CMV transmission has hindered vaccine development. Because young children shed high levels of computer virus in saliva and urine following postnatal contamination, they are major sources of transmission to pregnant women [4, 5]. Antibody may be capable of protecting against CMV contamination, based on findings of early phase clinical trials of CMV vaccines, including those based on the viral glycoprotein B (gB) [6]. Additionally, high avidity and functional CMV-specific immunoglobulin G (IgG) is usually transferred to the fetus, achieving levels higher than those in maternal plasma at delivery [7]. This passive immunity may explain why only 40% of breastfeeding infants given birth to to CMV-seropositive mothers are infected postnatally despite near uniform exposure to CMV in breast milk [8]. Thus, investigations of the protective role of passively acquired maternal IgG in infants may elucidate targets to interrupt CMV transmission between mothers and infants. In this hypothesis-generating study, we used a unique birth cohort of Ugandan infants and their CMV-seropositive mothers [9] to conduct a comprehensive analysis of the relationship between maternal CMV glycoproteinCspecific IgG specificities and functions and the risk of postnatal contamination. Since T cells are not transferred from your mother to fetus in significant figures, this approach investigates the role of IgG in postnatal transmission risk impartial of T-cell immunity. While our data did not demonstrate major differences in CMV-specific IgG binding or function between mother-infant pairs with or without postnatal acquisition at 6 months of age, we recognized a potential association with CMV gBCspecific IgG and delayed infant postnatal CMV acquisition. The data presented in this study support continued development of a vaccine that uses gB as an immunogen in a future, likely multitarget CMV vaccine. MATERIALS AND METHODS Study Population Plasma samples were collected as part of a previously explained cohort of 32 mother-infant JNJ-7706621 pairs [9]; maternal plasma was obtained at delivery, and infant plasma specimens were obtained at approximately 6 weeks of age. All subjects provided informed consent as approved by the human subjects protection committees JNJ-7706621 in Kampala, Uganda; Seattle, Washington; and Vancouver, Canada. Weekly oral swab specimens were collected for viral quantitative polymerase chain reaction (qPCR) screening, and the timing of postnatal CMV contamination in infants was defined by the onset of prolonged viral shedding in saliva and/or detection of viremia [9]. Three mother-infant pairs were excluded, with 2 excluded because congenital CMV was confirmed by saliva qPCR in the first week of life and 1 excluded because they were followed up for <6 months. Cell Culture Human retinal pigment epithelial cells (ARPE-19, ATCC CRL-2302) were produced in Dulbeccos altered Eagles medium with high glucose (Gibco 1195065), supplemented with 10% fetal calf serum (FCS), 2mM L-glutamine, 50 U/mL penicillin, and 50 g/mL streptomycin. Human lung fibroblasts (MRC-5, ATCC CCL-171) were produced in minimal essential medium (Gibco 11095080), supplemented with 10% FCS, 50 U/mL penicillin, and 50 g/mL streptomycin. Human monocyte cells (THP-1, ATCC TIB-202) were produced JNJ-7706621 in Roswell Park Memorial Institute 1640 medium (Gibco 11875119), supplemented with 10% FCS. Computer virus Production Bad(20 000 rpm) in a Beckman SW28 rotor for 1.5 hours. The computer virus pellet was resuspended in 10 mL of ARPE-19 growth medium adjusted to 0.2 M sucrose. Concentrated Badat 4oC, and incubated for 1 hour at 37oC with 5% CO2. Cells were then fixed with DPBS plus 1% formalin. AF647-positive cells were quantified using an LSR Fortessa (BD Biosciences). Data analysis was performed with FlowJo software. Natural Killer (NK) Cell Degranulation Assay NK cell.