Immune landscape of operational tolerance after allogeneic stem cell transplantation

the aim of this project was to characterize immune landscape of patients who underwent an allogeneic hematopoietic stem cell transplantation, one to two years after transplantation, and to characterize immune subsets associated with immune tolerance.
Conclusion:
We identified key immune cell subsets associated with operational tolerance after allogeneic stem cell transplantation. Cell events were acquired on the HELIOS mass cytometer (Fluidigm, Inc Canada) and CyTOF software version 6.7.1014 (Fluidigm, Inc Canada) at the “Plateforme de Cytométrie de la Pitié‐Salpetriere (CyPS).” An average of 500000 events was acquired per sample. Dual count calibration, noise reduction, cell length threshold between 10 and 150 pushes, and a lower convolution threshold equal to 10 were applied during acquisition. The standard mass cytometry files produced by HELIOS have been standardized using CyTOF v. 6.7.1014 software. This method normalizes each data to a global standard, called a log passport, determined for each equalization log (Fluidigm, Inc Canada) as recommended by the software developers. Once all samples are acquired, to control the normalization of each sample, they are again normalized together using MATLAB-based normalization software. Raw FCS files normalization was done using Fluidigm version 6.7.1014 Software and MatLab Normalizer version R2013 version 1, using Fluidigm® normalization beads.