title: Human regulatory T cells against minor histocompatibility antigens: ex vivo expansion for prevention of graft-versus-host disease.
abstract: Alloreactive donor T cells against host minor histocompatibility antigens (mHAs) cause graft-versus-host disease (GVHD) after marrow transplantation from HLA-identical siblings. We sought to identify and expand regulatory CD4 T cells (Tregs) specific for human mHAs in numbers and potency adequate for clinical testing. Purified Tregs from normal donors were stimulated by dendritic cells (DCs) from their HLA-matched siblings in the presence of interleukin 2, interleukin 15, and rapamycin. Male-specific Treg clones against H-Y antigens DBY, UTY, or DFFRY-2 suppressed conventional CD4 T cell (Tconv) response to the specific antigen. In the blood of 16 donors, we found a 24-fold (range, 8-fold to 39-fold) excess Tconvs over Tregs reactive against sibling mHAs. We expanded mHA-specific Tregs from 4 blood samples and 4 leukaphereses by 155- to 405-fold. Cultured Tregs produced allospecific suppression, maintained demethylation of the Treg-specific Foxp3 gene promoter, Foxp3 expression, and transforming growth factor β production. The rare CD4 T conv and CD8 T cells in the end product were anergic. This is the first report of detection and expansion of potent mHA-specific Tregs from HLA-matched siblings in sufficient numbers for application in human transplant trials.
authors.affiliation: Department of Blood and Marrow Transplantation.
authors.forename: Anandharaman
authors.initials: A
authors.lastname: Veerapathran
authors.forename: Joseph
authors.initials: J
authors.lastname: Pidala
authors.forename: Francisca
authors.initials: F
authors.lastname: Beato
authors.forename: Brian
authors.initials: B
authors.lastname: Betts
authors.forename: Jongphil
authors.initials: J
authors.lastname: Kim
authors.forename: Joel G
authors.initials: JG
authors.lastname: Turner
authors.forename: Marc K
authors.initials: MK
authors.lastname: Hellerstein
authors.forename: Xue-Zhong
authors.initials: XZ
authors.lastname: Yu
authors.forename: William
authors.initials: W
authors.lastname: Janssen
authors.forename: Claudio
authors.initials: C
authors.lastname: Anasetti
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D018920:Coculture Techniques; D005260:Female; D005434:Flow Cytometry; D006086:Graft vs Host Disease / Q000517:prevention & control*; D006801:Humans; D008297:Male; D015778:Minor Histocompatibility Antigens / Q000276:immunology*; D035781:Siblings; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000276:immunology*; D014184:Transplantation, Homologous
chemical_list: D015778:Minor Histocompatibility Antigens
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