Methods and compositions for the modification and delivery of lymphocytes

Abstract

Abstract The present disclosure provides methods and compositions for genetically modifying lymphocytes, for example T cells and / or NK cells. In some embodiments, the methods include reaction mixtures, and resulting cell formulations, that are created using whole blood, or a component thereof that is not a PBMC, and additionally comprise T cells and recombinant retroviral particles having polynucleotides that encode a CAR. In some embodiments, modified lymphocytes are reintroduced into a subject subcutaneously. In some embodiments, polynucleotides that provide T cells the ability to regulate cell survival and proliferation in response to binding to a CAR, are provided. Abstract WO 2021 / 042072 PCT / US2020 / 048843 3 / 31 FIG. 1E Primary Process Blood Collection Add Antibodies to Bind 110E Unwanted Cells Coarse Filter to Remove TNC Isolation Unwanted Cells 120E with Viral Particles Closed 130E Wash 140E Secondary Incubation Collect Cells ~37°C 150E Final Product in Infusible (optional) Bag, Syringe, or 200ECryopreserved in Vial 160E 20 26 20 61 65 16 J ul 2 02 6 w o 2 0 2 1 / 0 4 2 0 7 2 P C T / U S 2 0 2 0 / 0 4 8 8 4 3 2 0 2 6 2 0 6 1 6 5 1 6 J u l 2 0 2 6 O p t i o n a l P r o c e s s e s A d d A n t i b o d i e s t o B i n d 1 1 0 E 1 7 0 E C o a r s e F i l t e r t o R e m o v e T N C I s o l a t i o n 1 2 0 E 1 8 0 E " C o l d C o n t a c t i n g " o f T N C s C l o s e d 1 3 0 E W a s h 1 4 0 E ~ 3 7 ° C 1 9 0 E 1 5 0 E C o a r s e F i l t e r t o C a p t u r e C e l l A g g r e g a t e s 2 0 0 E 1 6 0 E

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