Nexvax2, but not placebo treatment in the RESET CeD Study was associated with significant reductions in gluten peptide-stimulated whole blood IL-2 and IFN-γ release, and CD4+ T cell proliferation. Delay in whole blood incubation of more than three hours from collection substantially reduced antigen-stimulated IL-2 and IFN-γ secretion. IL-2 and IFN-γ release in whole blood required more than 6 hours incubation. In contrast, whole blood CRA enabled detection of IL-2 and IFN-γ before and after gluten challenge. Only modest gluten peptide-stimulated IL-2 release could be detected without prior gluten challenge using PBMC. The pilot study showed that gluten peptides induced IL-2, IFN-γ and IL-10 release from PBMCs attributable to CD4+ T cells, but the PBMC CRA was substantially less sensitive than whole blood CRA. Subsequently, gluten-specific CD4+ T cells in blood were assessed in a subgroup of the RESET CeD Study participants who received Nexvax2 (maintenance dose 900 μg, n = 12) or placebo (n = 9). Dye-dilution proliferation in peripheral blood mononuclear cells (PBMC) was assessed, and IL-2, IFN-γ and IL-10 were measured by multiplex electrochemiluminescence immunoassay (ECL) after 24-hour gluten-peptide stimulation of whole blood or matched PBMC. Two participants with treated CD were assessed in a pilot study prior to and six days after a 3-day gluten challenge. We developed an enhanced CRA and used it in a phase 2 clinical trial (“RESET CeD”) of Nexvax2, a peptide-based immunotherapy for CD. Our aim was to develop a simple, but highly sensitive cytokine release assay (CRA) for gluten-specific CD4+ T cells that did not require patients to undergo a prior gluten challenge, and would be practical in large, multi-centre clinical trials. Improved blood tests assessing the functional status of rare gluten-specific CD4+ T cells are needed to effectively monitor experimental therapies for coeliac disease (CD). 6Department of Gastroenterology, The Royal Melbourne Hospital, Parkville, VIC, Australia.5Department of Gastroenterology, Alfred Hospital, Prahran, VIC, Australia.4Department of Gastroenterology, Box Hill Hospital, Box Hill, VIC, Australia.3ImmusanT, Inc., Cambridge, MA, United States.2Department of Medical Biology, University of Melbourne, Parkville, VIC, Australia.1Immunology Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.Brown 5, Suyue Wang 3, Evan Szymczak 3, Ruan Zhang 3, Kaela E.
0 kommentar(er)
