Production and Characterization of SIV-Specific CAR/CXCR5 T Cells

Area of Science:

• Immunology
• Gene Therapy
• Virology

Background:

• HIV infection establishes reservoirs in lymphoid follicles, particularly within T follicular helper cells.
• Current HIV therapies struggle to eradicate these viral reservoirs, necessitating novel treatment strategies.
• Chimeric antigen receptor (CAR) T cell therapy offers a potential approach to target and eliminate HIV-infected cells.

Purpose of the Study:

• To outline methods for producing HIV-specific CAR T cells engineered for follicular homing.
• To evaluate the in vitro and ex vivo functionality of these CAR T cells in a simian immunodeficiency virus (SIV) rhesus macaque model.
• To establish a preclinical framework for developing CAR T cell-based HIV immunotherapy.

Main Methods:

• Production of a retrovirus encoding an SIV-specific CAR and the follicular homing molecule CXCR5.
• Transduction of primary peripheral blood mononuclear cells (PBMCs) from rhesus macaques with the retrovirus.
• In vitro and ex vivo assays to assess T cell migration towards CXCL13 and into B cell follicles.
• Viral suppression assays to determine the antiviral activity of CAR T cells.

Main Results:

• Successful production of SIV-specific CAR/CXCR5-transduced T cells from rhesus macaque PBMCs.
• Demonstrated ability of CAR/CXCR5 T cells to migrate towards the CXCR5 ligand CXCL13 in vitro.
• Confirmed migration of transduced T cells into B cell follicles in ex vivo assays.
• Evidence of antiviral activity by CAR/CXCR5 T cells in viral suppression assays.

Conclusions:

• The described methods enable the production and functional evaluation of CAR T cells for HIV/SIV immunotherapy.
• Engineered T cells expressing CAR and CXCR5 show potential for targeting follicular HIV reservoirs.
• This preclinical model provides a pathway for developing effective CAR T cell therapies for HIV infection.