Oncotarget

Priority Research Papers:

Surrogate in vitro activation of innate immunity synergizes with interleukin-7 to unleash rapid antigen-driven outgrowth of CD4+ and CD8+ human peripheral blood T-cells naturally recognizing MUC1, HER2/neu and other tumor-associated antigens

Latha B. Pathangey, Dustin B. McCurry, Sandra J. Gendler, Ana L. Dominguez, Jessica E. Gorman, Girish Pathangey, Laurie A. Mihalik, Yushe Dang, Mary L. Disis and Peter A. Cohen _

PDF  |  HTML  |  Supplementary Files  |  How to cite  |  Order a Reprint

Oncotarget. 2017; 8:10785-10808. https://doi.org/10.18632/oncotarget.13911

Metrics: PDF 3340 views  |   HTML 6664 views  |   ?  


Abstract

Latha B. Pathangey1,*, Dustin B. McCurry1,*, Sandra J. Gendler1,2,3, Ana L. Dominguez1, Jessica E. Gorman1, Girish Pathangey1, Laurie A. Mihalik3, Yushe Dang4, Mary L. Disis4 and Peter A. Cohen2,3

1 Department of Biochemistry and Molecular Biology, Mayo Clinic, Scottsdale, AZ, USA

2 Department of Immunology, Mayo Clinic, Scottsdale, AZ, USA

3 Department of Hematology and Oncology, Mayo Clinic, Scottsdale, AZ, USA

4 Tumor Vaccine Group, Center for Translational Medicine in Women’s Health, University of Washington, Seattle, WA, USA

* Shared first authorship

Correspondence to:

Peter A. Cohen, email:

Sandra J. Gendler, email:

Mary L. Disis, email:

Keywords: PBMC, TLR agonists, GM-CSF, IL-7, adoptive therapy, MUC1

Received: October 07, 2016 Accepted: November 23, 2016 Published: December 11, 2016

Abstract

Effective adoptive immunotherapy has proved elusive for many types of human cancer, often due to difficulties achieving robust expansion of natural tumor-specific T-cells from peripheral blood. We hypothesized that antigen-driven T-cell expansion might best be triggered in vitro by acute activation of innate immunity to mimic a life-threatening infection. Unfractionated peripheral blood mononuclear cells (PBMC) were subjected to a two-step culture, first synchronizing their exposure to exogenous antigens with aggressive surrogate activation of innate immunity, followed by γ-chain cytokine-modulated T-cell hyperexpansion. Step 1 exposure to GM-CSF plus paired Toll-like receptor agonists (resiquimod and LPS), stimulated abundant IL-12 and IL-23 secretion, as well as upregulated co-stimulatory molecules and CD11c expression within the myeloid (CD33+) subpopulation. Added synthetic long peptides (>20aa) derived from widely expressed oncoproteins (MUC1, HER2/neu and CMVpp65), were reliably presented to CD4+ T-cells and cross-presented to CD8+ T-cells. Both presentation and cross-presentation demonstrated proteasomal and Sec61 dependence that could bypass the endoplasmic reticulum. Step 2 exposure to exogenous IL-7 or IL-7+IL-2 produced selective and sustained expansion of both CD4+ and CD8+ peptide-specific T-cells with a predominant interferon-γ-producing T1-type, as well as the antigen-specific ability to lyse tumor targets. Other γ-chain cytokines and/or combinations were initially proliferogenic, but followed by a contractile phase not observed with IL-7 or IL-7+IL-2. Regulatory T-cells were minimally propagated under these culture conditions. This mechanistically rational culture sequence, effective even for unvaccinated donors, enables rapid preparation of T-cells recognizing tumor-associated antigens expressed by the majority of human cancers, including pancreatic cancers, breast cancers and glioblastomas.


Creative Commons License All site content, except where otherwise noted, is licensed under a Creative Commons Attribution 3.0 License.
PII: 13911