title: Preparation of human ovarian cancer ascites-derived exosomes for a clinical trial.
abstract: Despite initial response to chemotherapy, at least 50% of ovarian cancer patients will relapse within 18 months. Progression-free survival is related to tumour infiltration with cytotoxic T lymphocytes (CTL). We recently demonstrated that CD8+ T cell responses to recall antigens improve following tumour response to chemotherapy. Vaccination designed to expand CTL, specific for tumour-associated antigens, may be a means of improving outcome. We are planning a clinical trial in advanced ovarian cancer patients undergoing chemotherapy using a combination of a Toll-like receptor 3 (TLR3) agonist and tumour-associated ascites-derived exosomes. Tumour-derived exosomes are a potential source of tumour antigens able to induce CD8+ T cell responses when loaded on mature dendritic cells (DC). DC maturation can be achieved with Toll-like receptor (TLR) agonists, such as the GMP-grade synthetic double stranded RNA, poly[I]:poly[C12U] (Ampligen) which is a TLR-3 agonist. Here, we describe the development of a method suitable for the preparation of GMP-grade exosomes from the ascites fluid of ovarian cancer patients, and the methods used for the molecular and immunological characterisation of these exosomes preceding their use in a clinical trial.
authors.affiliation: Cancer Services, Velindre NHS Trust, Cardiff, UK.
authors.forename: H
authors.initials: H
authors.lastname: Navabi
authors.forename: D
authors.initials: D
authors.lastname: Croston
authors.forename: J
authors.initials: J
authors.lastname: Hobot
authors.forename: A
authors.initials: A
authors.lastname: Clayton
authors.forename: L
authors.initials: L
authors.lastname: Zitvogel
authors.forename: B
authors.initials: B
authors.lastname: Jasani
authors.forename: R
authors.initials: R
authors.lastname: Bailey-Wood
authors.forename: K
authors.initials: K
authors.lastname: Wilson
authors.forename: Z
authors.initials: Z
authors.lastname: Tabi
authors.forename: M D
authors.initials: MD
authors.lastname: Mason
authors.forename: M
authors.initials: M
authors.lastname: Adams
mesh_terms: D001201:Ascites / Q000276:immunology*; D002986:Clinical Trials as Topic / Q000379:methods; D011992:Endosomes / Q000276:immunology / Q000637:transplantation*; D005260:Female; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D008722:Methods; D010051:Ovarian Neoplasms / Q000276:immunology / Q000628:therapy*; D051196:Toll-Like Receptor 3 / Q000819:agonists
chemical_list: C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3