To each PCR sample, 2 μl loading buffer was added, PD0325901 and the samples were ran for 30 min at 150V in gel electrophoresis of 2.5% agarose (Medionova) stained with ethidium bromide (EtBr) (Sigma-Aldrich,
Brøndby, Denmark). Medians and ranges are reported for continuous variables and percentages for categorical variables. Probabilities for overall survival and disease-free survival were calculated using the Kaplan–Meier estimator. All other outcomes used the cumulative incidence estimator. All outcomes were compared using a pointwise P-value at a specific point in time. Cox proportional hazards regression models were fit to the other outcomes. The proportional hazard assumption was assessed for each variable using a time-dependent approach. Variables used in the analysis include recipient age, Karnofsky performance score, use of ATG, disease, disease stage, stem cell source, GvHD prophylaxis, time from diagnosis to transplant for
CML, CMV matching, year of transplant, donor sex and number of donor pregnancies (Table 4). Stepwise model selection procedures were applied to build the models from the prognostic variables under consideration. We adopted a level of threshold (P-value <0.05) for variable selections. Each genetic marker was forced into the models that were built in the initial step and tested for association separately. Recipient genetic markers and donor genetic markers were treated separately in the analysis. Due to selleck inhibitor multiple testing, the P-values in the range 0.01–0.05 should be interpreted with caution test. For pairwise linkage disequilibrium analysis, the Lewontin’s D was used. The IL-7Rα genotype frequencies of patients and donors were comparable (Table 2) and corresponded to previously reported gene frequencies [10, 17]. The SNPs are in strong linkage disequilibrium (Table 3). In the univariate analysis, IL-7Rα rs1494558 was found to be associated with grades 2–4 aGVHD as well as cGVHD at 1 year,
the probability being highest in patients receiving transplants from donors with TT genotype (Table 4 and Fig. 1). A similar pattern was observed for IL-7Rα rs1494555, where the G allele was significantly associated with Etofibrate increased grades 2–4 aGVHD and cGVHD. By multivariate analysis, however, these associations were not significant. Neither rs1494558 nor rs1494555 was associated with overall survival or TRM (Table 5). By univariate and multivariate analysis, IL-7Rα rs6897932TT genotype of the donor was suggestive of an association with increased frequency of relapse (overall P = 0.015) compared with CC and CT donors (Fig. 2, Tables 4 and 5). The C allele was associated with increased risk of grades 3–4 aGVHD by univariate analysis (Table 4), but the association did not hold in the multivariate model (Table 5). No association was found between IL-7Rα rs6897932 genotypes and OS or TRM.