BackgroundHematopoietic stem cell transplantation (HSCT) is a well-established treatment modality for a variety of diseases. Immune reconstitution is an important event that determines outcomes. The immune recovery of T cells relies on peripheral expansion of mature graft cells, followed by differentiation of donor-derived hematopoietic stem cells. The formation of new T cells occurs in the thymus and as a byproduct, T cell receptor excision circles (TRECs) are released. Detection of TRECs by PCR is a reliable method for estimating the amount of newly formed T cells in the circulation and, indirectly, for estimating thymic function. The aim of this study was to determine the role of TREC quantitation in predicting outcomes of human leucocyte antigen (HLA) identical allogenic HSCT.MethodsThe study was conducted on 100 patients receiving allogenic HSCT from an HLA identical sibling. TREC quantification was done by real time PCR using a standard curve.ResultsTREC levels were inversely related to age (P=0.005) and were significantly lower in patients with malignant diseases than in those with benign diseases (P=0.038). TREC levels could predict relapse as an outcome but not graft versus host disease (GvHD) and infections.ConclusionAge and nature of disease determine the TREC levels, which are related to relapse.
FXYD3 is a Na/K-ATPase regulator which has been recently associated with different cancers development and progression; consequently, FXYD3 has been proposed in those as a potential therapeutic target. By contrast, no data are available concerning FXYD3 expression in hematological malignancies. In this study we aimed to assess FXYD3 gene expression in a large panel of B-cell derived lymphoid malignancies and to evaluate possible clinic-pathological correlations. Normal B-cell subsets served as control. We found that FXYD3 gene was not significantly modulated in normal B-cells. By contrast, BL, DLBCL, PMBCL, and PCM presented with a significant over-expression of the gene when compared to their cellular counterpart (p<0.0006). Interestingly, tumors characterized by higher FXYD3 expression presented with a significant enrichment in specific cellular functions and pathways, including NFkB pathway, WNT/B-catenin signaling, and MYC network, while FXYD3 levels also turned out to be directly related to PRDM1/BLIMP1. Finally, higher FXYD3 expression was not significantly associated with patients' survival in PCM and DLBCL, though a trend in favour of patients with lower expression was recorded in DLBCL cases. In conclusion, we unveiled FXYD3 gene over-expression in specific non-Hodgkin lymphoma subtypes and PCM, providing evidences of its involvement in their pathobiology. Future studies are needed to define its precise role.
Chimerism analysis is an important method for monitoring outcome of allogenic hematopoietic stem cell transplantation. Variable number tandem (VNTR) analysis is considered an informative technique to follow up chimerism state. The aim of the work is to study certain number of VNTRs to identify their potential value in the detection of chimerism in transplanted patients in conventional ablative transplants. Also to demonstrate the value of T cells subset analysis in detecting transplantation outcomes. This study included 17 pairs undergoing HSCT. Informative loci pre-transplantation using five VNTR loci and two gene loci were identified. After transplantation the informative loci were used to detect chimerism status. After DNA extraction from blood samples, amplification of VNTR loci was performed using a conventional PCR protocol. Extra sample post transplantation was collected and pre-treated with resetting technique in order to separate T cells, its product was subjected to DNA extraction then amplification of informative loci was done. Amplified product of DNA samples was run on 2% agarose gel stained using ethidium bromide. Fourteen recipients showed full done chimerism in both whole blood and T cells separated cells, one recipient died after HSCT, one recipient showed split chimerism and one pair failed to detect informative locus. VNTR analysis using a panel of five loci is suitable to detect state of chimerism after HSCT.
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