title: Metronomic cyclophosphamide enhances HPV16E7 peptide vaccine induced antigen-specific and cytotoxic T-cell mediated antitumor immune response.
abstract: In clinical trials, metronomic cyclophosphamide (CPA) is increasingly being combined with vaccines to reduce tumor-induced immune suppression. Previous strategies to modulate the immune system during vaccination have involved continuous administration of low dose chemotherapy, studies that have posed unique considerations for clinical trial design. Here, we evaluated metronomic CPA in combination with a peptide vaccine targeting HPV16E7 in an HPV16-induced tumor model, focusing on the cytotoxic T-cell response and timing of low dose metronomic CPA (mCPA) treatment relative to vaccination. Mice bearing C3 tumors were given metronomic CPA on alternating weeks in combination with immunization with a DepoVax vaccine containing HPV16E749-57peptide antigen every 3 weeks. Only the combination therapy provided significant long-term control of tumor growth. The efficacy of the vaccine was uncompromised if given at the beginning or end of a cycle of metronomic CPA. Metronomic CPA had a pronounced lymphodepletive effect on the vaccine draining lymph node, yet did not reduce the development of antigen-specific CD8+T cells induced by vaccination. This enrichment correlated with increased cytotoxic activity in the spleen and increased expression of cytotoxic gene signatures in the tumor. Immunity could be passively transferred through CD8+T cells isolated from tumor-bearing mice treated with the combinatorial treatment regimen. A comprehensive survey of splenocytes indicated that metronomic CPA, in the absence of vaccination, induced transient lymphodepletion marked by a selective expansion of myeloid-derived suppressor cells. These results provide important insights into the multiple mechanisms of metronomic CPA induced immune modulation in the context of a peptide cancer vaccine that may be translated into more effective clinical trial designs.
authors.affiliation: Immunovaccine Inc. ; Halifax; Nova Scotia, Canada ; Department of Microbiology & Immunology; Dalhousie University ; Halifax; Nova Scotia, Canada.
authors.forename: Genevieve M
authors.initials: GM
authors.lastname: Weir
authors.affiliation: Immunovaccine Inc. ; Halifax; Nova Scotia, Canada ; Department of Biology; Dalhousie University ; Halifax; Nova Scotia, Cananda.
authors.forename: Olga
authors.initials: O
authors.lastname: Hrytsenko
authors.affiliation: Immunovaccine Inc. ; Halifax; Nova Scotia, Canada ; Department of Microbiology & Immunology; Dalhousie University ; Halifax; Nova Scotia, Canada.
authors.forename: Marianne M
authors.initials: MM
authors.lastname: Stanford
authors.affiliation: Ontario Institute for Cancer Research ; Toronto; Ontario, Canada.
authors.forename: Neil L
authors.initials: NL
authors.lastname: Berinstein
authors.affiliation: Immunovaccine Inc. ; Halifax; Nova Scotia, Canada ; Department of Microbiology & Immunology; Dalhousie University ; Halifax; Nova Scotia, Canada.
authors.forename: Mohan
authors.initials: M
authors.lastname: Karkada
authors.affiliation: Department of Microbiology & Immunology; Dalhousie University ; Halifax; Nova Scotia, Canada ; Division of Hematopathology; Queen Elizabeth II Health Sciences Centre ; Nova Scotia, Canada.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Liwski
authors.affiliation: Immunovaccine Inc. ; Halifax; Nova Scotia, Canada.
authors.forename: Marc
authors.initials: M
authors.lastname: Mansour
keywords: CPA, cyclophosphamide; CTL, cytotoxic T lymphocyte; CTLA-4, cytotoxic T lymphocyte-associated protein 4; DPX, DepoVax; HPV, human papilloma virus; HPV16; IFNγ, interferon γ; MDSC, myeloid-derived suppressor cells; PD-1/PDCD1, programmed cell death 1; PO, per os (oral); Treg, regulatory T cell; cancer; checkpoint inhibitors; mCPA, metronomic low dose CPA; metronomic cyclophosphamide; sbCPA, single bolus low dose CPA; translational; vaccine