CCR5-targeted allogeneic gamma-delta CD19 chimeric antigen receptor T cells for HIV-associated B cell-malignancy immunotherapy

Ángel Ramírez-Fernández^#^{1

2

3

4

5}, Alexander J Dimitri^#^{6

7

8

9

10}, Fang Chen⁷, Robert Bartoszek^{6

7

8

9

10}, Gregory M Chen^{6

7

8

9

10}, Laura Córdoba-Espejo^{7

8

9}, Yuqi Zhou^{6

7}, Yun-Hsin Tang⁷, Chien-Ting Lin⁷, Reyes Acosta^{6

7}, John Scholler⁷, Guido Ghilardi^{7

8

11

12}, Patrizia Porazzi^{7

8

11

12}, Mireia Pellicer¹³, Núria Profitós-Pelejà¹³, Stefan K Barta^{11

12}, Anne Chew^{7

9}, Julie K Jadlowsky^{7

9}, Vanessa E Gonzalez^{7

8}, Donald L Siegel^{7

8

9}, Bruce L Levine^{7

8

9}, Gaël Roué¹³, Marco Ruella^{7

8

11

12}, Michael T Lotze¹⁴, Carl H June^{7

8

9

10}, James L Riley^{6

7

8

9}, Joseph A Fraietta^{15

16

17

18

19}

Affiliations

¹ Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
² Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
³ Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
⁴ Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
⁵ Parker Institute for Cancer Immunotherapy, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
⁶ Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁷ Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁸ Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁹ Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹⁰ Parker Institute for Cancer Immunotherapy, University of Pennsylvania, Philadelphia, PA, USA.
¹¹ Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA, USA.
¹² Division of Hematology-Oncology, Department of Medicine, Hospital of the University of Pennsylvania, Philadelphia, PA, USA.
¹³ Lymphoma Translational Group, Josep Carreras Leukaemia Research Institute (IJC), Badalona, Barcelona, Spain.
¹⁴ Hillman Cancer Center, University of Pittsburgh, Pittsburgh, PA, USA.
¹⁵ Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.
¹⁶ Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.
¹⁷ Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.
¹⁸ Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.
¹⁹ Parker Institute for Cancer Immunotherapy, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.

^# Contributed equally.

PMID: 41120570
DOI: 10.1038/s41551-025-01527-0

CCR5-targeted allogeneic gamma-delta CD19 chimeric antigen receptor T cells for HIV-associated B cell-malignancy immunotherapy

Ángel Ramírez-Fernández et al. Nat Biomed Eng. 2025.

. 2025 Oct 21.

doi: 10.1038/s41551-025-01527-0. Online ahead of print.

Authors

Affiliations

¹ Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
² Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
³ Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
⁴ Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
⁵ Parker Institute for Cancer Immunotherapy, University of Pennsylvania, Philadelphia, PA, USA. angel.ramirez-fernandez@pennmedicine.upenn.edu.
⁶ Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁷ Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁸ Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
⁹ Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹⁰ Parker Institute for Cancer Immunotherapy, University of Pennsylvania, Philadelphia, PA, USA.
¹¹ Lymphoma Program, Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA, USA.
¹² Division of Hematology-Oncology, Department of Medicine, Hospital of the University of Pennsylvania, Philadelphia, PA, USA.
¹³ Lymphoma Translational Group, Josep Carreras Leukaemia Research Institute (IJC), Badalona, Barcelona, Spain.
¹⁴ Hillman Cancer Center, University of Pittsburgh, Pittsburgh, PA, USA.
¹⁵ Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.
¹⁶ Center for Cellular Immunotherapies, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.
¹⁷ Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.
¹⁸ Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.
¹⁹ Parker Institute for Cancer Immunotherapy, University of Pennsylvania, Philadelphia, PA, USA. jfrai@upenn.edu.

^# Contributed equally.

PMID: 41120570
DOI: 10.1038/s41551-025-01527-0

Abstract

Immune-based cell therapy offers a promising approach to cancer treatment. While autologous chimeric antigen receptor (CAR) T cells have shown success, production is time-consuming, costly and patient specific. Gamma-delta (γδ) T cells are promising for 'off-the-shelf' CAR T cell therapy. However, clinical translation of γδ CAR T cells is hampered by low frequency, resistance to genetic manipulation and advanced differentiation after expansion, limiting therapeutic feasibility. Here we demonstrate a method for in vitro activation and expansion of peripheral blood γδ T cells, facilitating high rates of gene editing and efficient CAR integration. Using artificial antigen-presenting cells, we produce minimally differentiated, highly functional γδ CAR T cells. By targeting a US Food and Drug Administration-approved CD19 CAR to the CCR5 locus, we generate CCR5-deficient γδ CD19 CAR T cells (γδ CCR5KI-CAR19), which demonstrated resistance to HIV-mediated depletion and robust antitumour responses against B cell lymphoma and leukaemia. γδ CCR5KI-CAR19 T cells enable the immunotherapy of HIV-associated B cell malignancies. These studies provide preclinical evidence supporting large-scale development of potent allogeneic γδ CAR T cells for diverse immunotherapies.

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Conflict of interest statement

Competing interests: A.C.: co-founder and equity holder in Tmunity Therapeutics. D.L.S.: founder’s equity and licensed IP with Verismo Therapeutics, Vetigenics and Chimeric Therapeutics. B.L.L.: consultancy and advisory roles with Terumo, GSK and Kite; co-founder and equity holder in Tmunity Therapeutics (acquired by Kite) and Capstan Therapeutics; advisory board memberships with Avectas, Capstan (Chair), Immuneel, Immusoft, In8bio, Ori Biotech, Oxford Biomedica, Thermo Fisher Pharma Services and UTC Therapeutics; member of the Alliance for Cancer Gene Therapy Board of Directors. M.R.: holds patents related to CD19 CAR T cells; consultant for NanoString, Bristol Myers Squibb, GlaxoSmithKline, Scailyte, Bayer and AbClon; research funding from AbClon, NanoString, Oxford NanoImaging, viTToria Biotherapeutics, CURIOX and Beckman Coulter; scientific founder of viTToria Biotherapeutics. C.H.J.: receives royalties from Novartis and Kite paid to the University of Pennsylvania; scientific co-founder and equity holder in Capstan Therapeutics, Dispatch Biotherapeutics and BlueWhale Bio; board member of AC Immune; scientific advisory roles with various companies, including BluesphereBio, Cabaletta, Carisma, Cartography, Cellares, Cellcarta, Celldex, Danaher, Decheng, ImmuneSensor, Kite, Poseida, Verismo, Viracta and WIRB-Copernicus Group. J.L.R.: received grants from Tmunity/Kite outside of the submitted work; co-founder of Tmunity Therapeutics and BlueWhale Bio with monetary compensation and equity. J.A.F.: holds patents and intellectual property in T cell-based cancer immunotherapy with royalties; receives funding from Tmunity Therapeutics and Danaher Corporation; consultancy with Retro Biosciences; scientific advisory board memberships with Cartography Bio, Shennon Biotechnologies Inc., CellFe Biotech, OverT Bio, Inc., and Tceleron Therapeutics, Inc. All other authors declare no competing interests. G.R.: received grants from Onconova Therapeutics outside of the submitted work.

References

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CCR5-targeted allogeneic gamma-delta CD19 chimeric antigen receptor T cells for HIV-associated B cell-malignancy immunotherapy

Affiliations

CCR5-targeted allogeneic gamma-delta CD19 chimeric antigen receptor T cells for HIV-associated B cell-malignancy immunotherapy

Authors

Affiliations

Abstract

Conflict of interest statement

References

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