Journal Article

CD33 Delineates Two Functionally Distinct NK Cell Populations Divergent in Cytokine Production and Antibody-Mediated Cellular Cytotoxicity

Bennstein, Sabrina B., Möker, Nina, Rühl, Heiko, Uhrberg, Markus, Pfeifer, Rita, Reusing, Sarah B., Quadflieg, Melissa, Oldenburg, Johannes, Wels, Winfried S., Balz, Vera, Nitsche, Marcus, Oberbeck, Sebastian, Fischer, Johannes C., Zhang, Congcong, Heinrichs, Stefan, Hanenberg, Helmut, Oberoi, Pranav, Hejazi, Maryam, Cramer, Sophie, Hoerster, Keven, Brossart, Peter, Horn, Peter A., Babor, Florian

2022

Overview

The generation and expansion of functionally competent NK cells in vitro is of great interest for their application in immunotherapy of cancer. Since CD33 constitutes a promising target for immunotherapy of myeloid malignancies, NK cells expressing a CD33-specific chimeric antigen receptor (CAR) were generated. Unexpectedly, we noted that CD33-CAR NK cells could not be efficiently expanded in vitro due to a fratricide-like process in which CD33-CAR NK cells killed other CD33-CAR NK cells that had upregulated CD33 in culture. This upregulation was dependent on the stimulation protocol and encompassed up to 50% of NK cells including CD56 dim NK cells that do generally not express CD33 in vivo . RNAseq analysis revealed that upregulation of CD33 + NK cells was accompanied by a unique transcriptional signature combining features of canonical CD56 bright (CD117 high , CD16 low ) and CD56 dim NK cells (high expression of granzyme B and perforin). CD33 + NK cells exhibited significantly higher mobilization of cytotoxic granula and comparable levels of cytotoxicity against different leukemic target cells compared to the CD33 − subset. Moreover, CD33 + NK cells showed superior production of IFNγ and TNFα, whereas CD33 − NK cells exerted increased antibody-dependent cellular cytotoxicity (ADCC). In summary, the study delineates a novel functional divergence between NK cell subsets upon in vitro stimulation that is marked by CD33 expression. By choosing suitable stimulation protocols, it is possible to preferentially generate CD33 + NK cells combining efficient target cell killing and cytokine production, or alternatively CD33 − NK cells, which produce less cytokines but are more efficient in antibody-dependent applications.

• Publisher: Frontiers Media SA,Frontiers Media S.A
• Subject: Antibodies / Antibody-Dependent Cell Cytotoxicity - immunology / Antigens / CD33-CAR / CD56 Antigen - immunology / CD56 Antigen - metabolism / Cell culture / Cells / Cells, Cultured / Chimeric antigen receptors / cytokine production and cytotoxicity / Cytokines / Cytokines - immunology / Cytokines - metabolism / Cytotoxicity / Cytotoxicity, Immunologic - immunology / Flow Cytometry - methods / Gene Expression Profiling - methods / Granzyme B / Humans / Immunology / Immunotherapy / K562 Cells / Killer Cells, Natural - immunology / Killer Cells, Natural - metabolism / Leukemia / Ligands / Malignancy / Natural killer cells / NK cell / NK cell expansion / Penicillin / Perforin / Proto-Oncogene Proteins c-kit - genetics / Proto-Oncogene Proteins c-kit - immunology / Proto-Oncogene Proteins c-kit - metabolism / Protocol / Receptors, Chimeric Antigen - immunology / Receptors, Chimeric Antigen - metabolism / Receptors, IgG - genetics / Receptors, IgG - immunology / Receptors, IgG - metabolism / RNAseq analysis / Sialic Acid Binding Ig-like Lectin 3 - genetics / Sialic Acid Binding Ig-like Lectin 3 - immunology / Sialic Acid Binding Ig-like Lectin 3 - metabolism / Software / Up-Regulation / γ-Interferon