PMC7204010_01
Changes in the patient's skin before and after ssCART-19 cells infusion. a, Left lower limb skin. b, Skin biopsy from left lower limb and staining by HE. c, Facial skin. d, Back skin
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Fig1
However, after 3 cycles of consolidation treatment, the skin nodules (Fig. 1a, c, d) and the swelling of the scrotum reappeared (Fig. 2c).  The pathology of skin biopsy was consistent with B lymphoblastic lymphoma/ALL infiltration (Fig. 1b). However, 1 week after infusion the patient's skin nodules disappeared, and only the chromatosis remained (Fig. 1a, c, d).
The summary of medication protocol, clinical and laboratory parameters relative to the timing of CART therapy. a, Chemotherapy for lymphpcyte depletion include fludarabine and cyclophosphamide. ssCART-19 cells were infused at a split-dose of 10% on day 01, 30% on day 02 and 60% on day 03(total 5 x 106/kg). b, Expressions of blasts in bone marrow and skin were detected by flow cytometry before and after ssCART-19 treatment. c, Testiclar size was measured by ultrasound respectively on the -5 day (before CAR-T), on + 4 day (during CAR-T), + 7 day (after CAR-T), + 37 day (before HSCT) and most recently (after HSCT). d, Related hematological toxicity after ssCART-19 treatment. e, The trends of the patient's temperature in degrees centigrade ( C) per 24-h period. IL-6(pg/ml), IFN(pg/ml) and hsCRP(mg/l) concentrations are shown in lines during CART therapy. f, The expansion levels of ssCART-19 cells in the peripheral blood (PB) were monitored by qPCR on each day
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Fig2
Flow cytometry analysis of skin biopsy showed 93.1% of blast proportion with immature B cell immunophenotyping profile (CD10+: 99.9%, CD22+: 99.8%, CD19+: 90.9%, CD20 + -: 97.9%, CD38+: 99%, CD34 + -: 73.9%)(Fig. 2b). Fludarabine 30 mg/m2/day and Cyclophosphamide 250 mg/m2/day. Infection-related examinations including procalcitonin, spot test using T lymphocytes, 1, 3 - beta glucan test, galactomannan test and chest CT scanning, were performed and showed negative results. Two days after conditioning, the patient received ssCART-19 cells infusion at a total dose of 5 x 106 cells/kilogram for 3 consecutive days (Fig. 2a). Four days after infusion, the patient suffered from lower swelling in skin nodules and left testicle, accompanied by high fever. Biopsy showed few lymphocytes infiltration but no leukemia cells, which was confirmed by flow cytometry (Fig. 2b). Besides, the ultrasound examination revealed shirked testicles (Fig. 2c). The levels of IL-6, IFN-gammaand hsCRP were elevated following ssCART-19 cells infusion; they were consistent with the changes in body temperature, and declined to basal level spontaneously in 1 week (Fig. 2e). CRS in grade 1 was considered only due to discomfort related to fevers (Fig. 2e) and no CRES was detected. The proliferation of ssCART-19 cells detected by the real-time quantitative PCR was noted on day + 6, and sustained on a high level for several days (Fig. 2f). The patient had granulocytopenia without thrombocytopenia or anemia after infusion (Fig. 2d).