In a phase 1 trial, bispecific CAR T cells targeting CD19 and CD22 exhibit clinical activity and low toxicity in patients with large B cell lymphoma and B cell acute lymphoblastic leukemia jayspiegel25 MackallLab StanfordCancer ASH22
. Samples were stained with a panel backbone first. Subsequently, samples were split into two for either phenotyping of T cell subsets or exhaustion markers ; all samples were prepared and washed as described above. Analysis for this and the previous two sections were performed in FlowJo v10.5.3 .This section describes the methods used for the intracellular cytokine panel in Fig.
. All NALM6 lines were divided into two and stained with either CD19 or CD22 antibody and prepared and washed as described above.This section describes the methods used for the intracellular cytokine panel in Fig.. Clinical CAR T products were thawed and rested overnight before incubation in coculture assays with the NALM6 lines described above. During this 6–7 h coculture at 37 °C, cells were coincubated with monensin and CD107a.
. CD19-22.BB.z cells were used as the positive batch control for the daily staining experiments. At least 10cells were acquired unless restricted by the number of cells isolated from 8 ml of whole blood. The assay limit of detection for CAR T cells was calculated as 1 in 10DNA was extracted from whole blood using the QIAamp DNA Mini Kit at baseline and on days 7, 14, 21, 28, 90 and 180 postCAR infusion.
. ctDNA was measured from plasma extracted from blood obtained in EDTA tubes at pre-, 0, 7, 14, 21, 56 and 90 d postCD19-22.BB.z-CAR infusion.Descriptive statistics were enumerated by median and IQR for continuous variables and counts and percentages for categorical variables. Fisher’s exact and McNemar’s tests evaluated the association between categorical variables and Spearman correlation was used for the association between two random continuous variables.
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