Post-infusion CAR TReg cells identify patients resistant to CD19-CAR therapy - Nature Medicine

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Single-cell proteomic profiling of circulating CART cells in patients treated with CD19-CAR shows that CD4+Helios+ CAR Treg cells on day 7 after infusion are associated with progressive disease & less severe neurotoxicity in patients with LBCL.

lymphocytes, as defined by the forward and side scatter gates. Absolute CAR T cell numbers were calculated by multiplying the percentages of CAR T cells among lymphocytes by the absolute lymphocyte count obtained on the same day. The assay limit of detection was calculated as 1 in 10DNA was extracted from 2–5 × 10PBMCs using QIAmp DNA Blood Mini Kit .

. In brief, cryopreserved PBMCs were thawed into 9 ml of cell culture medium supplemented with 25 U ml, resuspended in 10 ml of CCM, filtered through a 70-µm strainer and counted., cisplatin was reconstituted to 100 mM in DMSO and incubated at 37 °C for 3 days to prepare a stock solution, which was then stored in aliquots at −20 °C. Cell pellets were resuspended in 1 ml of PBS containing 0.

 

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Thank you for highlighting our work! We found that this CAR Treg population is Helios+ FOXP3+, and quantification based on either marker led to nearly identical results.

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