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: Anandharaman Veerapathran, Joseph Pidala, Francisca Beato, Brian Betts, Jongphil Kim, Joel G Turner, Marc K Hellerstein, Xue-Zhong Yu, William Janssen, Claudio Anasetti
• Journal: Blood (Blood) Vol. 122 Issue 13 Pg. 2251-61 (Sep 26 2013) ISSN: 1528-0020 [Electronic] United States
• PMID: 23908471 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Minor Histocompatibility Antigens

Topics

• Cell Culture Techniques (methods)
• Coculture Techniques
• Female
• Flow Cytometry
• Graft vs Host Disease (prevention & control)
• Humans
• Male
• Minor Histocompatibility Antigens (immunology)
• Siblings
• T-Lymphocytes, Regulatory (cytology, immunology)
• Transplantation, Homologous