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Research Article: ?-Galactosylceramide-expanded virtual memory CD8 + T cells confer protection against a broad range of pathogens

Date Published: 2026-05-22

Abstract:
?-Galactosylceramide (?-GalCer) treatment of animals is a well-established model to investigate the beneficial roles of invariant natural killer T (iNKT) cells in anti-infection and anti-tumor immunity. We previously found that ?-GalCer treatment expands not only iNKT cells but also a group of antigen-inexperienced memory-like innate virtual memory CD8 + T (CD8 + T VM ) cells. With the bystander protective potential of CD8 + T VM cells, their roles in ?-GalCer-mediated immunity against infection remain largely unexplored. Here, we reported that ?-GalCer treatment conferred protection against systemic, enteric, and pulmonary bacterial infections, as well as pulmonary viral infection. This protection persisted after iNKT cells had diminished and coincided with sustained expansion of CD8 + T VM cells. Adoptive transfer of ?-GalCer-expanded CD8 + T VM cells one day before infection significantly decreased bacterial and viral burdens. Mechanistically, ?-GalCer treatment activated iNKT cells to produce IL-4, which subsequently expanded the number of CD8 + T VM cells that were capable of producing TNF-? and IFN-?. But the ability of each individual CD8 + T VM cell to produce these cytokines was not enhanced. Lung-localized CD8 + T VM cells were the primary contributors to ?-GalCer-expanded CD8 + T VM cells in the lung. ?-GalCer-expanded CD8 + T VM cells can reduce pulmonary viral titer in the host in a TNF-?-dependent manner in the absence of IFN-? signaling. Together, these findings describe an immune cascade in which ?-GalCer-activated iNKT cells orchestrate the expansion and function of CD8 + T VM cells through IL-4 production, reveal previously unrecognized roles of CD8 + T VM cells as downstream effectors in ?-GalCer-mediated immunotherapeutic effects, and highlight the protective potential of CD8 + T VM cells in host defense against diverse pathogens, thus providing a novel approach to developing new therapeutics for combating not only various infectious diseases but also tumors.

Introduction:
?-Galactosylceramide (?-GalCer), a marine sponge-derived glycolipid, is a natural ligand of invariant natural killer T (iNKT) cells and is commonly used to specifically activate iNKT cells through the presentation by MHC-class I-like molecule CD1d on antigen-presenting cells (APCs) ( 1 ). Upon ?-GalCer stimulation, iNKT cells can rapidly proliferate ( 2 ) and produce a broad range of T H 1, T H 2, and T H 17 cytokines ( 3 ), which subsequently activate dendritic cells (DCs), natural killer (NK) cells, and…

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