3. METHODS
CAR-T cell expansion in PB was monitored by multiparameter flow
cytometry through detection of labeled CD19 in T cells (Human CD19
Protein®) and by quantitative PCR using the HIV /COBAS® AmpliPrep/COBAS®
TaqMan® HIV-1 Test, v2.0 (Roche, Switzerland) following manufacturer’s
instructions.
Tumor samples from core needle biopsies were processed using QIAamp DNA
Tissue Kit (Qiagen, Germany) to detect CAR-T cells.
Plasma cytokine levels (IL-6, IL-15, IL-1β, and GM-CSF) were determined
with last generation ELISA multiplex (Ella®, ProteinSimple®).
Written informed consent was obtained in compliance with our
institutional review board and the Declaration of Helsinki.