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, Peter A Cohen

Research output: Contribution to journalArticle

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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.

Original languageEnglish (US)
Pages (from-to)10785-10808
Number of pages24
JournalOncotarget
Volume8
Issue number7
DOIs
StatePublished - 2017

Fingerprint

Interleukin-7
Neoplasm Antigens
Innate Immunity
Blood Cells
T-Lymphocytes
Antigens
Interleukin-2
resiquimod
Neoplasms
Cross-Priming
Breast Neoplasms
Cytokines
Peptide T
Interleukin-23
Adoptive Immunotherapy
Toll-Like Receptors
Oncogene Proteins
Regulatory T-Lymphocytes
Glioblastoma
Interleukin-12

Keywords

  • Adoptive therapy
  • GM-CSF
  • IL-7
  • MUC1
  • PBMC
  • TLR agonists

ASJC Scopus subject areas

  • Oncology

Cite this

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

In: Oncotarget, Vol. 8, No. 7, 2017, p. 10785-10808.

Research output: Contribution to journalArticle

Pathangey, Latha B. ; McCurry, Dustin B. ; Gendler, Sandra J ; Dominguez, Ana L. ; Gorman, Jessica E. ; Pathangey, Girish ; Mihalik, Laurie A. ; Dang, Yushe ; Disis, Mary L. ; Cohen, Peter A. / 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. In: Oncotarget. 2017 ; Vol. 8, No. 7. pp. 10785-10808.
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