We identified 5 DA-associated potential regulatory SNPs. Four variants exhibited effects on gene regulation (ATF rs11571537, CDH17 rs2446824 and rs2513789, and TACR1 rs2287231). A fifth variant (FAM71A rs147978008) showed nonrisk allele preferential binding to H1 histones. These results demonstrate that many DA-associated genetic variants likely act by modulating gene regulation.

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RATIONALE: Genetic markers of susceptibility for DA are needed to identify exposed workers at highest risk. We have used GWAS and Next Generation Sequencing (NGS) methodologies to identify gene variants associated with specific inhalation challenge confirmed DA. METHODS: Workers with occupational asthma caused by diisocyanates (i.e., HDI, MDI, or TDI) were recruited at occupational health clinics in Canada and Spain. Based on a prior GWAS study, 14 DA associated loci and their flanking regions were selected for sequencing (6,996,180 base pairs). NGS was performed in 91 workers with confirmed DA (cases) and compared to 293 subjects with sequencing data from the 1,000 genomic (1KG) control data set. RESULTS: NGS analysis of DA cases compared to 1KG controls yielded 143 significant SNP associations with DA (Chi-Squared-log 10 P< 3.0) (range 3.006-5.504), of which all but one SNP were located in intronic regions. Transcriptomic analysis was carried out for the top 70 DA associated SNPs using available transcription factor (TF) datasets from relevant lungderived tissues and cell lines, including ChIP-seq datasets for TFs, regulatory histone marks, and DNase-seq (open chromatin). SNPs were ranked based on the number of datasets they overlap and the biologic relevance of the associated SNPs. The top 22 SNPs identified for further study are contained in 7 loci: CDH17 (10), ATF3 (6), FAM71A (2), PITPNC1, TACR1, ZBTB16, LOC101929565. CONCLUSIONS: This study identified DA-associated SNPs linked with transcription binding sites. The possible roles of these variants on gene expression and protein function are under investigation.

RATIONALE: Adoptive immunotherapy using virus-specific T-lymphocytes (VSTs) has been successful in treating viral infections after hematopoietic stem cell transplantation (HSCT). Use of partially HLAmatched VSTs from third-party donors allow ''off the shelf'' therapy for viral infections. We hypothesize that third-party VST will be effective and safe in patients with primary immunodeficiency disorders (PID). METHODS: VSTs were cultured from healthy donors via viral pepmix stimulation and expansion. Patients received third party VST infusion before or after HSCT for treatment of CMV, EBV, or adenovirus. Antiviral immunity was assessed via IFNg ELISpot, IFNg capture flow cytometry, and TCRb sequencing. Patients were followed for 45 days following infusion for toxicities and for up to 12 months for antiviral immune reconstitution. RESULTS: Seven patients with PID received 9 VST products at a dose of 2 x 10E7 cells/m2. Four patients had undergone HSCT, while three patients were treated before HSCT due to persistent viral infections. None of the patients developed GVHD, though transient flaring of viral hepatitis occurred in two patients after infusion. Antiviral responses were seen in 5 of 7 patients. Of the pre-HSCT patients, two infants with SCID and cidofovir-refractory adenovirus each received 2 VST infusions and cleared their infections. Expansion of virus-specific T-cells was detectable by IFNg ELIspot or gamma capture in 3 of 5 responding patients. VST persistence was also confirmed by TCRb sequencing in one patient. CONCLUSIONS: Third-party VST are safe in patients with PID, and may be a valuable salvage therapy for the treatment of viral infections before or after HSCT.

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