Selective expression of latency-associated peptide (LAP) and IL-1 receptor type I/II (CD121a/CD121b) on activated human FOXP3+ regulatory T cells allows for their purification from expansion cultures.
Although adoptive transfer of regulatory T cells (Foxp3(+) Tregs) has proven to be efficacious in the prevention and treatment of autoimmune diseases and graft-versus-host disease in rodents, a major obstacle for the use of Treg immunotherapy in humans is the difficulty of obtaining a highly purified preparation after ex vivo expansion. We have identified latency-associated peptide (LAP) and IL-1 receptor type I and II (CD121a/CD121b) as unique cell-surface markers that distinguish activated Tregs from activated FOXP3(-) and FOXP3(+) non-Tregs. We show that it is feasible to sort expanded FOXP3(+) Tregs from non-Tregs with the use of techniques for magnetic bead cell separation based on expression of these 3 markers. After separation, the final product contains greater than 90% fully functional FOXP3(+) Tregs. This novel protocol should facilitate the purification of Tregs for both cell-based therapies as well as detailed studies of human Treg function in health and disease.
Tran
Dat Q
DQ
Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA. dtran@niaid.nih.gov
Andersson
John
J
Hardwick
Donna
D
Bebris
Lolita
L
Illei
Gabor G
GG
Shevach
Ethan M
EM
D015415:Biomarkers; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D051858:Forkhead Transcription Factors; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D011956:Receptors, Cell Surface; D053573:Receptors, Interleukin-1 Type I / Q000032:analysis*; D053574:Receptors, Interleukin-1 Type II / Q000032:analysis*; D050378:T-Lymphocytes, Regulatory / Q000166:cytology*
D015415:Biomarkers; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; C508592:IL1R2 protein, human; D011956:Receptors, Cell Surface; D053573:Receptors, Interleukin-1 Type I; D053574:Receptors, Interleukin-1 Type II