Effects of transmembrane region variability on cell surface expression and allorecognition of HLA-DP3

Crivello, Pietro; Lauterbach, Nina; Zito, Laura; Sizzano, Federico; Toffalori, Cristina; Marcon, Jessica; Curci, Laura; Mulder, Arend; Wieten, Lotte; Zino, Elisabetta; Voorter, C. E. M.; Tilanus, Marcel G.J.; Fleischhauer, Katharina

doi:10.1016/j.humimm.2013.04.014

Cited by 10 publications

(11 citation statements)

References 33 publications

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“…WT and mutant HLA-DP expression by BLCL was driven by a bidirectional lentiviral expression platform, as previously described [24,25], inducing coordinate expression of transgenic HLA-DPB1 and the DLNGFR reporter. After magnetic bead purification for reporter gene expression, cell surface expression levels of transgenic DLNGFR or HLA-DPB1 were quantitatively assessed by flow cytometric determination of Molecules of Equivalent Soluble Fluorochrome [26] using SPHERO Rainbow Calibration Particles (Spherotech, Lake Forest, IL) [25].…”

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

“…After magnetic bead purification for reporter gene expression, cell surface expression levels of transgenic DLNGFR or HLA-DPB1 were quantitatively assessed by flow cytometric determination of Molecules of Equivalent Soluble Fluorochrome [26] using SPHERO Rainbow Calibration Particles (Spherotech, Lake Forest, IL) [25].…”

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

“…Isolation and Characterization of CD4 þ T Cells Alloreactive to HLA-DPB1*09:01 PBMC from responder-stimulator pairs matched for 9 to 10/10 of the HLA-A, B, C, DRB1, and DQB1 alleles but mismatched for HLA-DPB1, were used for the set-up of 1-way mixed lymphocyte reactions (MLR) as previously described [25]. PBMC were from a patient before transplantation or at the time of HSCT allograft rejection [27] or from 5 different HLA-typed healthy subjects.…”

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

“…For complete HLA typing of the PBMC, see typing of the respective BLCL in Table S1. After 2 rounds of stimulation, T cells were either cloned by limiting dilution or left uncloned and maintained by repeated restimulations, as described [25]. Allospecificity of T cell clones was established by IFN-g ELISpot assays according to standard protocols [28,29] and …”

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

“…Alloreactive TCR-Vb subfamilies in uncloned T cell cultures were analyzed by flow-cytometric CD137 upregulation assays as described [25,30,31], using appropriately labeled anti-CD4 and anti-CD137 mAb in combination with mAb for different TCR-Vb subfamilies. A list of alloreactive T cells used, along with their TCR-Vb subfamilies and their allospecificity, is included as Table S2.…”

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

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The Impact of Amino Acid Variability on Alloreactivity Defines a Functional Distance Predictive of Permissive HLA-DPB1 Mismatches in Hematopoietic Stem Cell Transplantation

Crivello

Zito

Sizzano

et al. 2015

Biology of Blood and Marrow Transplantation

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104

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A major challenge in unrelated hematopoietic stem cell transplantation (HSCT) is the prediction of permissive HLA mismatches, ie, those associated with lower clinical risks compared to their nonpermissive counterparts. For HLA-DPB1, a clinically prognostic model has been shown to be matching for T cell epitope (TCE) groups assigned by cross reactivity of T cells alloreactive to HLA-DPB1∗09:01; however, the molecular basis of this observation is not fully understood. Here, we have mutated amino acids (aa) in 10 positions of HLA-DPB1∗09:01 to other naturally occurring variants, expressed them by lentiviral vectors in B cell lines, and quantitatively measured allorecognition by 17 CD4(+) T cell effectors from 6 unrelated individuals. A significant impact on the median alloresponse was observed for peptide contact positions 9, 11, 35, 55, 69, 76, and 84, but not for positions 8, 56, and 57 pointing away from the groove. A score for the "functional distance" (FD) from HLA-DPB1∗09:01 was defined as the sum of the median impact of polymorphic aa in a given HLA-DPB1 allele on T cell alloreactivity. Established TCE group assignment of 23 alleles correlated with FD scores of ≤0.5, 0.6 to 1.9 and ≥2 for TCE groups 1, 2, and 3, respectively. Based on this, prediction of TCE group assignment will be possible for any given HLA-DPB1 allele, including currently 367 alleles encoding distinct proteins for which T cell cross reactivity patterns are unknown. Experimental confirmation of the in silico TCE group classification was successfully performed for 7 of 7 of these alleles. Our findings have practical implications for the applicability of TCE group matching in unrelated HSCT and provide new insights into the molecular mechanisms underlying this model. The innovative concept of FD opens new potential avenues for risk prediction in unrelated HSCT.

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Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

Section: Monoclonal Antibodies and Flow Cytometrymentioning

confidence: 99%

See 3 more Smart Citations

The Impact of Amino Acid Variability on Alloreactivity Defines a Functional Distance Predictive of Permissive HLA-DPB1 Mismatches in Hematopoietic Stem Cell Transplantation

Crivello

Zito

Sizzano

et al. 2015

Biology of Blood and Marrow Transplantation

Self Cite

111

104

View full text Add to dashboard Cite

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HapLogic: A Predictive Human Leukocyte Antigen–Matching Algorithm to Enhance Rapid Identification of the Optimal Unrelated Hematopoietic Stem Cell Sources for Transplantation

Dehn

Setterholm

Buck

et al. 2016

Biology of Blood and Marrow Transplantation

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The search for a suitable human leukocyte antigen (HLA)-matched unrelated adult stem cell donor (URD) or umbilical cord blood unit (UCB) is a complex process. The National Marrow Donor Program (NMDP) developed a search algorithm known as HapLogic, which is currently provided within the NMDP Traxis application. The HapLogic algorithm has been in use since 2006 and has advanced URD/UCB HLA-matching technology. The algorithm has been shown to have high predictive accuracy, which can streamline URD/UCB selection and drive efficiencies in the search process to the benefit of the stem cell transplantation community. Here, we describe the fundamental components of the NMDP matching algorithm, output, validation, and future directions.

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Uncommon HLA alleles identified by hemizygous ultra-high Sanger sequencing: haplotype associations and reconsideration of their assignment in the Common and Well-Documented catalogue

et al. 2016

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Effects of transmembrane region variability on cell surface expression and allorecognition of HLA-DP3

Cited by 10 publications

References 33 publications

The Impact of Amino Acid Variability on Alloreactivity Defines a Functional Distance Predictive of Permissive HLA-DPB1 Mismatches in Hematopoietic Stem Cell Transplantation

The Impact of Amino Acid Variability on Alloreactivity Defines a Functional Distance Predictive of Permissive HLA-DPB1 Mismatches in Hematopoietic Stem Cell Transplantation

HapLogic: A Predictive Human Leukocyte Antigen–Matching Algorithm to Enhance Rapid Identification of the Optimal Unrelated Hematopoietic Stem Cell Sources for Transplantation

Uncommon HLA alleles identified by hemizygous ultra-high Sanger sequencing: haplotype associations and reconsideration of their assignment in the Common and Well-Documented catalogue

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