Peripheral vaccination-induced brain-resident memory CD8+ T cells durably protect mice against intracranial malignancy
Madison R. Mix, Cassie M. Sievers, Mariah Hassert, Shravan Kumar Kannan, Lecia L. Pewe, Sunny C. Huang, Rui He, Cori E. Fain, Mohammad Heidarian, Lisa S. Hancox, Sahaana A. Arumugam, Terry G. Beltz, Fang Jin, Aaron J. Johnson, Calvin S. Carter, Noah S. Butler, Aliasger K. Salem, Vladimir P. Badovinac, John T. Harty
Madison R. Mix, Cassie M. Sievers, Mariah Hassert, Shravan Kumar Kannan, Lecia L. Pewe, Sunny C. Huang, Rui He, Cori E. Fain, Mohammad Heidarian, Lisa S. Hancox, Sahaana A. Arumugam, Terry G. Beltz, Fang Jin, Aaron J. Johnson, Calvin S. Carter, Noah S. Butler, Aliasger K. Salem, Vladimir P. Badovinac, John T. Harty
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Research Article Immunology Neuroscience Oncology

Peripheral vaccination-induced brain-resident memory CD8+ T cells durably protect mice against intracranial malignancy

Abstract

Primary and metastatic brain tumors exhibit resistance to immunotherapies that demonstrate efficacy in peripheral cancer settings. While many immunotherapies aim to enhance CD8+ T cell infiltration and functionality in established tumors, identification of neoantigens support emerging immunopreventative tactics against brain cancer. Functionally potent tissue-resident memory CD8+ T cells (TRM) can be generated in the brain following peripheral infection or vaccination. However, the ability of brain TRM to prevent intracranial malignancy remains unknown. Here, mice were seeded with tumor-specific or bystander brain TRM via peripheral infection prior to depletion of circulating memory T cells (TCIRCM) and subsequent brain tumor challenge. Tumor-specific brain TRM durably protected mice against intracranial malignancy even in the absence TCIRCM. These brain TRM persisted in tumor-surviving mice and protected against a second antigen-matched challenge. Importantly, a translationally-relevant mRNA-lipid nanoparticle (LNP) vaccine phenocopied peripheral infection-induced outcomes, generating functional brain TRM that controlled tumor growth. Altogether, this work points to the utility of brain TRM in cancer immunoprevention, supporting the development of antitumor mRNA-LNP vaccines to bolster brain immunity.

Authors

Madison R. Mix, Cassie M. Sievers, Mariah Hassert, Shravan Kumar Kannan, Lecia L. Pewe, Sunny C. Huang, Rui He, Cori E. Fain, Mohammad Heidarian, Lisa S. Hancox, Sahaana A. Arumugam, Terry G. Beltz, Fang Jin, Aaron J. Johnson, Calvin S. Carter, Noah S. Butler, Aliasger K. Salem, Vladimir P. Badovinac, John T. Harty

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Figure 5

Peripheral mRNA-LNP vaccination generates tumor-specific brain TRM.

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Peripheral mRNA-LNP vaccination generates tumor-specific brain TRM. (A) ...
(A) Construct design. Following the 5’ untranslated region (UTR), a cleavage-resistant ubiquitin (Ub-A76), a flexible linker (FL), and the coding sequences for TRP1455–463, TRP2180–188, GP10025–33, and OVA257–264 flanked by AAY proteasomal cleavage sites were encoded. (B) Experimental design. Thy1.2 C57BL/6N mice were adoptively transferred with 104 Thy1.1 OT-I i.v. and immunized one day later with 5 μg UbMel-OVA mRNA-LNP vaccine (Vx) i.m. or i.v. with boosting 28 days later. (C) Number of OT-I/ml blood across time. (D) Number of OT-I in the spleen, cervical draining lymph nodes (cDLN; number of cells divided by number of cDLN retrieved), and IV– brain. (E) Frequency of IFN-γ+ and (F) CD107a+ OT-I in the IV– brain following 5–6-hour ex vivo 1 μM OVA257–264 peptide stimulation. (G) TRM-associated marker expression among OT-I in i.m. and (H) i.v. vaccinated hosts. (I) Thy1.1+ OT-I and CD31+ vasculature in white-gray matter junction regions of i.m. vaccinated hosts. Scale bar: 200 μm. (J) Representative H2-Db TRP1455–463, H2-Kb TRP2180–188, and H2-Db GP10025–33 tetramer (Tet) staining in the spleen. (K) Number of TRP1455–463- and (L) GP10025–33-specific CD8+ T cells in the spleen and IV– brain. Experiments in (A–F and J–L) show concatenated data from 2 independent experiments with n = 7–20 mice per group total. Experiments (G and H) show representative data from 1 of 2 independent experiments with n = 4–5 mice per group. Experiments (I) show representative images from n = 3–4 mice. Statistical significance was determined by Student’s t test or 1-way ANOVA with Tukey’s multiple comparison’s test. Graphs show the mean ± SEM with each symbol representing 1 mouse. Individual P values are noted on respective graphs or are otherwise summarized as: *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Graphical illustrations were created using BioRender (https://biorender.com).