For tagging SNPs we used an r2 0.8. factor receptor (PDGFRA) and integrins were also selected based on their role in binding gB. Specific SNPs in TLR7 and IKBKE (inhibitor of nuclear factor kappa-B kinase subunit epsilon) were associated with antibody responses to gB vaccine. Homozygous carriers of the minor allele at four SNPs in TLR7 showed higher vaccination-induced antibody responses to gB compared to heterozygotes or homozygotes for the common allele. SNP rs1953090 in IKBKE was associated with changes in antibody level from second to third dose of vaccine; homozygotes for the minor allele exhibited lower antibody responses while homozygotes for the major allele showed increased responses over time. Conclusions These data contribute to our understanding of the immunogenetic mechanisms underlying variations in the immune response to CMV vaccine. strong class=”kwd-title” Keywords: Cytomegalovirus, Toll-like receptors, single nucleotide polymorphisms, glycoprotein B vaccine Background Infection with CMV is common in humans, causing severe morbidity and mortality in congenitally-infected newborns and in immunocompromised patients [1-3]. The importance of CMV as the leading infectious cause of mental retardation and deafness in children has been emphasized by its categorization by the Institute of Medicine as a level I vaccine candidate . The rationale for developing a CMV vaccine is based on clinical and animal studies showing that immunity to CMV reduces the frequency and severity of disease [5,6]. In addition, animal studies demonstrated that immunization with subunit vaccines prevented disease and transplacental transmission of CMV [5-7]. Two recent phase II clinical trials with glycoprotein B (gB)-MF59 led to major enthusiasm and hope for the future success of CMV vaccine. The first was performed in young women recruited on postpartum wards , and showed 50% efficacy in preventing maternal CMV infection. Analysis of antibody levels to gB among vaccine Triptolide (PG490) recipients revealed that all women developed antibodies to gB although the levels and kinetics of antibody responses varied. The second study recruited patients from the kidney/liver transplant waiting list, and showed that antibody titers against gB were significantly increased one month after the second injection in patients given the vaccine compared with those given placebo, and that antibody titers to gB pretransplant correlated inversely with the duration of viremia, and the need for therapy with ganciclovir after transplant . Data from human studies suggest that single nucleotide polymorphisms (SNPs) in immune response genes may influence severity of infections and response to vaccinations such as rubella, measles and hepatitis B [9-16]. Toll-like receptors (TLR) play a key role in the innate immune system and have been implicated in infectious and autoimmune processes . CMV gB and glycoprotein gH (gH) associate with and activate TLR2/1, mediating an initial signal transduction pathway leading to upregulation of NF-kB and SP-1 [18,19]. In liver transplant recipients TLR2 R753Q SNP was associated with CMV replication and disease . The Triptolide (PG490) successful gB vaccine trial in young women provided us with a unique opportunity to determine whether antibody responses to gB vaccine were influenced by SNPs in TLR genes. Methods Study population The study cohort included healthy women who were enrolled in the CMV vaccine after obtaining written informed consent . Women were screened on the postpartum wards, and those Triptolide (PG490) who were negative for antibody to CMV were invited to participate in the clinical trial. Study participants received a fixed dose of vaccine consisting of recombinant CMV envelope glycoprotein Triptolide (PG490) B (0.02 mg) with MF59 adjuvant (13.25 mg). The Johns Hopkins University School of Medicine and University of Alabama Institutional Review Boards granted SIRT1 approval for this study. Antibody assays Antibody to CMV gB was measured using an Enzyme-linked immunoabsorbent assay (ELISA) . The vaccine antigen, a recombinant gB molecule from Towne CMV (provided by Sanofi Pasteur, Marcy L’Etoile, France) was used. SNP selection Using a candidate gene approach the following genes were selected: TLRs and associated intracellular signaling molecules: TLR1-4, TLR6, TLR7, TLR9, TLR10, JUN, MYD88, IKBKE, CHUK (IKK), NF-KB1, CD14, MXD3 (MAD3), MAPK8 (JNK1), MAPK14, MAP3K7 (TAK1), LY96 (MD2), TRAF6, IRAK1, IRAK4, TBK1, TICAM1 (TRIF) and IRF3. In addition, PDGFRA, PDGFRB and integrin alpha Triptolide (PG490) V and integrin B1 were selected based on data showing their role in binding gB [22,23]. We identified tagging SNPs within the genes and in the 10 kb flanking each side of the genes, using the LDselect algorithm in individuals.