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Chainonthee W.,TU Dresden | Chainonthee W.,DKMS Life Science Laboratory GmbH | Bottcher G.,DKMS Life Science Laboratory GmbH | Gagne K.,Laboratoire dHistocompatibilite et dImmunogenetique | And 6 more authors.
Tissue Antigens | Year: 2010

Molecular analysis of genetic polymorphism for clinical or research purposes may be compromised by genomic DNA of limited quality and quantity. In this study, we have successfully tested the feasibility of using whole genome amplification (WGA) to allow genotyping for killer cell immunoglobulin-like receptor (KIR) genes and human leucocyte antigen (HLA)-C KIR ligand dimorphism on HLA-C. WGA was achieved by multiple displacement amplification (MDA) using bacteriophage φ29 polymerase. For KIR genotyping, a revised sequence-specific primer polymerase chain reaction protocol consisting of 23 primer pairs was used avoiding hitherto undetected cross-priming involving KIR2DL1, KIR2DS1, KIR3DL1 and KIR3DS1 alleles. Similarly, MDA-amplified genomic DNA was analyzed for the detection of the HLA-C KIR ligand groups C1 and C2, based on the amino acid K/N dimorphism in position 80. © 2010 John Wiley & Sons A/S.

Petersdorf E.W.,Fred Hutchinson Cancer Research Center | Petersdorf E.W.,University of Washington | Gooley T.A.,Fred Hutchinson Cancer Research Center | Malkki M.,Fred Hutchinson Cancer Research Center | And 24 more authors.
Blood | Year: 2014

Life-threatening graft-versus-host disease (GVHD) limits the use of HLA-C-mismatched unrelated donors in transplantation. Clinicians lack criteria for donor selection when HLA-C-mismatched donors are a patient's only option for cure. We examined the role for HLA-Cexpressionlevels to identify permissible HLA-Cmismatches.Themedian fluorescence intensity, a proxy of HLA-Cexpression,was assigned to each HLA-C allotype in 1975 patients and their HLA-C-mismatched unrelated transplant donors. The association of outcome with the levelofexpressionof patients'anddonors'HLA-Callotypeswasevaluated inmultivariable models. Increasing expression level of the patient's mismatched HLA-C allotype was associated with increased risks of grades III to IV acute GVHD, nonrelapse mortality, and mortality. Increasing expression level among HLA-C mismatches with residue 116 or residue 77/80 mismatching was associated with increased nonrelapse mortality. The immunogenicity of HLA-C mismatches in unrelated donor transplantation is influenced by the expression level of the patient's mismatched HLA-C allotype. HLA-C expression levels provide newinformation onmismatches that should be avoided and extend understanding of HLA-C-mediated immune responses in human disease. © 2014 by The American Society of Hematology.

Le Bourgeois A.,University of Nantes | Labopin M.,HOpitalSaint Antoine | Guillaume T.,University of Nantes | Delaunay J.,University of Nantes | And 17 more authors.
Experimental Hematology | Year: 2014

Our main objective was to determine new factors associated with engraftment and single-unit predominance after double umbilical cord blood (UCB) allogeneic stem-cell transplantation. Engraftment occurred in 78% of cases in this retrospective study including 77 adult patients. Three-year overall survival, disease-free survival, relapse incidence, and nonrelapse mortality were 55±6%, 44±6%, 33±5%, and 23±4%, respectively. In multivariate analysis, Human herpesvirus 6 reactivation during aplasia (hazard ratio [HR]=2.63; 95% confidence interval [CI]: 1.64-4.17; p<0.001), younger recipient age (<53years) (HR=1.97; 95% CI: 1.16-3.35; p = 0.012), and lower human leukocyte antigenmatching between the two units (3 of 6 or 4 of 6) (HR=2.09; 95% confidence interval: 1.22-3.59; p = 0.013) were the three factors independently associated with graft failure. Also, factors independently predicting the losing UCB unit were younger age of the UCB unit (odds ratio [OR]=1.01; 95% CI: 1-1.02; p = 0.035), lower CD34+ cell dose contained in the UCB unit (≤0.8×105/kg) (OR=2.55; 95% CI: 1.05-6.16; p= 0.04), and presence of an ABO incompatibility between the UCB unit and the recipient (OR=2.53; 95% CI: 1.15-5.53; p = 0.02). Thus, Human herpesvirus 6 reactivation during aplasia, lower unit-unit human leukocyte antigenmatching, and younger UCB unit age, as new unfavorable predictive factors, may represent new parameters to take into account afterdouble UCB allogeneic stem-cell transplantation in adults. These results need to beconfirmed prospectively, as they may influence unit selections and patient outcomes. © 2014 ISEH - International Society for Experimental Hematology.

Gagne K.,University of Nantes | Gagne K.,Laboratoire dHistocompatibilite et dImmunogenetique | Willem C.,University of Nantes | Legrand N.,University of Nantes | And 9 more authors.
European Journal of Immunology | Year: 2013

NK-cell functions are regulated by many activating and inhibitory receptors including KIR3DL1. Extensive allelic polymorphism and variability in expression can directly alter NK-cell phenotype and functions. Here we investigated the KIR3DL1+ NK-cell repertoire, taking into account the allelic KIR3DL1/S1 polymorphism, KIR3DL1 phenotype, and function. All 109 studied individuals possessed at least one KIR3DL1 allele, with weak KIR3DL1*054, or null alleles being frequently present. In KIR3DL1high/null individuals, we observed a bimodal distribution of KIR3DL1+ NK cells identified by a different KIR3DL1 expression level and cell frequency regardless of a similar amount of both KIR3DL1 transcripts, HLA background, or KIR2D expression. However, this bimodal distribution can be explained by a functional selection following a hierarchy of KIR3DL1 receptors. The higher expression of KIR3DL1 observed on cord blood NK cells suggests the expression of the functional KIR3DL1*004 receptors. Thus, the low amplification of KIR3DL1high, KIR3DL1*004 NK-cell subsets during development may be due to extensive signaling via these two receptors. Albeit in a nonexclusive manner, individual immunological experience may contribute to shaping the KIR3DL1 NK-cell repertoire. Together, this study provides new insight into the mechanisms regulating the KIR3DL1 NK-cell repertoire. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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