An increasing number of gene and cell therapies have been authorized for different types of cancer and non-malignant diseases. However, novel cell therapies with advanced therapy medicinal products (ATMPs) are difficult to implement, particularly for HIV⁺ patients. We aim to address this unmet medical need in the context of CAR-T cells by establishing dedicated processes for ATMP manufacture. In addition, to-be-infused T cells will be protected from de novo HIV infection by CCR5-gene editing. We previously established efficient production of (i) CD19-CAR-T cells and (ii) CCR5ᵏᵒ T cells at the CliniMACS Prodigy. Now, we are developing a combined process for lentiviral CAR transfer and knockout of the HIV co-receptor CCR5 by mRNA electroporation of our TALE nuclease, CCR5-Uco-hetTALEN. We have successfully performed three manufacturing runs at the CliniMACS Prodigy to produce CCR5-edited CD19-CAR-T cells on a GMP-compliant, clinical scale. We produced up to 3.9x10⁹ viable T cells containing 37-55 % functional CD19-CAR-T cells, with an overall CCR5-editing rate of 42-54%. Furthermore, the third process was successfully performed under the addition of the antiretroviral substance saquinavir to supress viral replication during manufacturing of HIV⁺ cells.

In summary, we have developed a GMP-compliant manufacturing process for CCR5-edited CAR-T cells under antiretroviral suppression, which might help to accelerate the translation of cellular immunotherapies, namely CAR-T cells for HIV⁺ patients in the future.