title: Toll-like receptor agonist therapy can profoundly augment the antitumor activity of adoptively transferred CD8(+) T cells without host preconditioning.
abstract: BACKGROUND
  Lymphodepletion enhances adoptive T cell transfer (ACT) therapy by activating the innate immune system via microbes released from the radiation-injured gut. Microbial components, such as LPS, are key mediators of total body irradiation (TBI) enhancement, but our ability to strategically use these toll-like receptor (TLR) agonists to bolster the potency of T cell-based therapies for cancer remains elusive. Herein, we used TLR4 agonist LPS as a tool to address how and when to use TLR agonists to effectively improve cancer immunotherapy.
  
  
  METHODS
  To determine the mechanisms of how innate immune activation via lymphodepletion potentiated antitumor T cell immunity, we utilized the pmel-1 melanoma mouse model. B16F10-bearing mice were preconditioned with 5Gy TBI and given a tripartite ACT therapy (consisting of transferred pmel-1 CD8(+) T cells, vaccination with fowlpox encoding gp100, and IL-2) along with TLR4 agonist LPS. The timing of LPS administration and the requirement of individual components of the tripartite therapy were evaluated based on tumor growth and the phenotype of recovered splenocytes by flow cytometry. We also evaluated the role of non-toxic and clinically used TLR4 and TLR9 agonists-monophosphoryl lipid A (MPL) and CpG Oligodeoxynucleotide (CpG ODN), respectively- for ACT therapy.
  
  
  RESULTS
  Here we report that while exogenous administration of LPS was able to enhance adoptively transferred CD8(+) T cells' tumor destruction, LPS treatment alone did not replace individual components of the tripartite ACT regimen, or obviate TBI. Moreover, we found that sequentially administering LPS during or one day prior to ACT therapy compromised tumor regression. In contrast, administering LPS after ACT potentiated the antitumor effectiveness of the regimen, thereby supporting the expansion of transferred tumor-specific CD8(+) T cells over host CD4(+) T cells. We also found that non-toxic TLR agonists MPL and CpG potentiated the antitumor activity of infused CD8(+) T cells. Finally, TBI was no longer needed to regress tumors in mice who were depleted of host CD4(+) T cells, given a tripartite ACT regimen and then treated with low dose LPS.
  
  
  CONCLUSIONS
  Collectively, our results identify how and when to administer TLR agonists to augment T cell-based immunotherapy in the absence or presence of host preconditioning for treatment of advanced malignancies. Our findings have clinical implications for the design of next generation immune-based therapies for patients with cancer.
authors.affiliation: Microbiology and Immunology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425 USA.
authors.forename: Michelle H
authors.initials: MH
authors.lastname: Nelson
authors.affiliation: Microbiology and Immunology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425 USA.
authors.forename: Jacob S
authors.initials: JS
authors.lastname: Bowers
authors.affiliation: Microbiology and Immunology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425 USA.
authors.forename: Stefanie R
authors.initials: SR
authors.lastname: Bailey
authors.affiliation: Microbiology and Immunology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425 USA.
authors.forename: Marshall A
authors.initials: MA
authors.lastname: Diven
authors.affiliation: Microbiology and Immunology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425 USA.
authors.forename: Caroline W
authors.initials: CW
authors.lastname: Fugle
authors.affiliation: Center for Cancer Research, National Cancer Institute (NCI), National Institutes of Health (NIH), Bethesda, MD 20892 USA.
authors.forename: Andrew D M
authors.initials: AD
authors.lastname: Kaiser
authors.affiliation: Center for Cancer Research, National Cancer Institute (NCI), National Institutes of Health (NIH), Bethesda, MD 20892 USA.
authors.forename: Claudia
authors.initials: C
authors.lastname: Wrzesinski
authors.affiliation: Microbiology and Immunology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425 USA.
authors.forename: Bei
authors.initials: B
authors.lastname: Liu
authors.affiliation: Center for Cancer Research, National Cancer Institute (NCI), National Institutes of Health (NIH), Bethesda, MD 20892 USA.
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: Restifo
authors.affiliation: Microbiology and Immunology, Hollings Cancer Center, Medical University of South Carolina, Charleston, SC 29425 USA.
authors.forename: Chrystal M
authors.initials: CM
authors.lastname: Paulos
keywords: Adoptive immunotherapy; CD8+ T lymphocytes; Innate immunity; Total body Irradiation