II DQ8 antigen (at a MFI of 2313) and still expressed antibodies against this antigen at 3, 6 and 12 months, albeit at a lower reactivity (MFIs: 760, 659 and 772, respectively). A third patient developed DSA against both HLA I B44, only detected after 6 months at a low MFI (607), and HLA II DQ2 at 3 months (MFI:1027) with a further increase at 6 months (3889). IFN-γ Elispot The low level of immunosuppression was confirmed by the anti-viral response assessed by the ex IFNγ Elispot assay with immunodominant viral peptides. Three patients presented at baseline a vivo positive T cell response against peptides derived from CMV or a pool of viral peptides (CEF) and this response did not decrease during follow-up while 2 other patients did not respond at baseline and this lack of T cell reactivity persisted during the follow-up. Detection of circulating donor-derived deoxyribonucleic acid (DNA) ESCORT patients’ plasma DNAs and exogenous ESC-derived cardiovascular progenitors were analyzed as described above. Polymorphisms used to detect exogenous ESC-derived cardiovascular progenitor DNA were those present in the cell line and absent in patients. MET p.Ile 377Ile c.1131C>T and SMAD4 p.Phe362Phe c.1086 T>C were cell-line specific and tested for all patients. For 2 cases, the EGFR p.Gln787Gln c.2366 G>A was added as both patients were homozygotes for the G allele. One sample was not informative due to low coverage depth. Our previous data showed that allelic ratios of > 0.005 (0.5%) could be considered as significant alterations and that sequencing data showing alterations at lower frequencies had to be considered as sequencing artefacts. In this study, no plasma showed a cell line-specific allele ratio of more than 0.0018 (0.18%, MET ESCORT 5.2). References
II DQ8 antigen (at a MFI of 2313) and still expressed antibodies against this antigen at 3, 6 and 12 months, albeit at a lower reactivity (MFIs: 760, 659 and 772, respectively). A third patient developed DSA against both HLA I B44, only detected after 6 months at a low MFI (607), and HLA II DQ2 at 3 months (MFI:1027) with a further increase at 6 months (3889). IFN-γ Elispot The low level of immunosuppression was confirmed by the anti-viral response assessed by the ex vivo IFNγ Elispot assay with immunodominant viral peptides. Three patients presented at baseline a positive T cell response against peptides derived from CMV or a pool of viral peptides (CEF) and this response did not decrease during follow-up while 2 other patients did not respond at baseline and this lack of T cell reactivity persisted during the follow-up. Detection of circulating donor-derived deoxyribonucleic acid (DNA) ESCORT patients’ plasma DNAs and exogenous ESC-derived cardiovascular progenitors were analyzed as described above. Polymorphisms used to detect exogenous ESC-derived cardiovascular progenitor DNA were those present in the cell line and absent in patients. MET p.Ile 377Ile c.1131C>T and SMAD4 p.Phe362Phe c.1086 T>C were cell-line specific and tested for all patients. For 2 cases, the EGFR p.Gln787Gln c.2366 G>A was added as both patients were homozygotes for the G allele. One sample was not informative due to low coverage depth. Our previous data showed that allelic ratios of > 0.005 (0.5%) could be considered as significant alterations and that sequencing data showing alterations at lower frequencies had to be considered as sequencing artefacts. In this study, no plasma showed a cell line-specific allele ratio of more than 0.0018 (0.18%, MET ESCORT 5.2). References
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