title: Measuring human T cell responses in blood and gut samples using qualified methods suitable for evaluation of HIV vaccine candidates in clinical trials.
abstract: The next generation of candidate HIV vaccines include replicating vectors selected for tropism to mucosal sites, where an efficacious T cell response will be required to limit T cell replication and HIV associated CD4 T cell loss. To fully assess immunogenicity of such candidates, there is a need to develop robust quality controlled analysis of gut derived HIV specific CD8+ T-cell responses. Despite obvious challenges in obtaining sufficient amounts of tissue, the highly compartmentalised nature of the mucosal immune responses, requires the assessment of CD8 T cells isolated directly from local tissue before any conclusions regarding the induction of mucosal responses are made. Here we describe the optimisation and subsequent qualification of a qualitative and quantitative polychromatic flow cytometry assay to assess antigen specific CD8+ T cells isolated from the gut, using samples from HIV positive and negative volunteers. Internal quality controls monitored over time, combined with the use of quality gating and standard operating procedures were used to demonstrate the generation of robust and reliable data.
authors.affiliation: International AIDS Vaccine Initiative Human Immunology Laboratory (IAVI HIL), London, UK.
authors.forename: Harry
authors.initials: H
authors.lastname: Kaltsidis
authors.forename: Hannah
authors.initials: H
authors.lastname: Cheeseman
authors.forename: Jakub
authors.initials: J
authors.lastname: Kopycinski
authors.forename: Ambreen
authors.initials: A
authors.lastname: Ashraf
authors.forename: Michelle Cashin
authors.initials: MC
authors.lastname: Cox
authors.forename: Lorna
authors.initials: L
authors.lastname: Clark
authors.forename: Insiyah
authors.initials: I
authors.lastname: Anjarwalla
authors.forename: Len
authors.initials: L
authors.lastname: Dally
authors.forename: Phillip
authors.initials: P
authors.lastname: Bergin
authors.forename: Aggeliki
authors.initials: A
authors.lastname: Spentzou
authors.forename: Chris
authors.initials: C
authors.lastname: Higgs
authors.forename: Frances
authors.initials: F
authors.lastname: Gotch
authors.forename: Brian
authors.initials: B
authors.lastname: Gazzard
authors.forename: Raul
authors.initials: R
authors.lastname: Gomez
authors.forename: Peter
authors.initials: P
authors.lastname: Hayes
authors.forename: Peter
authors.initials: P
authors.lastname: Kelleher
authors.forename: Dilbinder K
authors.initials: DK
authors.lastname: Gill
authors.forename: Jill
authors.initials: J
authors.lastname: Gilmour
mesh_terms: D016915:AIDS Vaccines / Q000276:immunology*; D000328:Adult; D000368:Aged; D001706:Biopsy; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007413:Intestinal Mucosa / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D013601:T-Lymphocytes / Q000276:immunology*; D055815:Young Adult
chemical_list: D016915:AIDS Vaccines