A clinical-scale selective allodepletion approach for the treatment of HLA-mismatched and matched donor-recipient pairs using expanded T lymphocytes as antigen-presenting cells and a TH9402-based photodepletion technique.
Selective allodepletion is a strategy to eliminate host-reactive donor T cells from hematopoietic stem cell allografts to prevent graft-versus-host disease while conserving useful donor immune functions. To overcome fluctuations in activation-based surface marker expression and achieve a more consistent and effective allodepletion, we investigated a photodepletion process targeting activation-based changes in p-glycoprotein that result in an altered efflux of the photosensitizer TH9402. Expanded lymphocytes, generated using anti-CD3 and IL-2, were cocultured with responder cells from HLA-matched or -mismatched donors. Optimal results were achieved when cocultured cells were incubated with 7.5 muM TH9402, followed by dye extrusion and exposure to 5 Joule/cm(2) light energy at 5 x 10(6) cells/mL. In mismatched stimulator-responder pairs, the median reduction of alloreactivity was 474-fold (range, 43-fold to 864-fold) compared with the unmanipulated responder. Third-party responses were maintained with a median 1.4-fold (range, 0.9-fold to 3.3-fold) reduction. In matched pairs, alloreactive helper T-lymphocyte precursors were reduced to lower than 1:100 000, while third-party responses remained higher than 1:10 000. This establishes a clinical-scale process capable of highly efficient, reproducible, selective removal of alloreactive lymphocytes from lymphocyte transplant products performed under current Good Manufacturing Practice. This procedure is currently being investigated in a clinical trial of allotransplantation.
Mielke
Stephan
S
Allotransplantation Section, Hematology Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892-1202, USA.
Nunes
Raquel
R
Rezvani
Katayoun
K
Fellowes
Vicki S
VS
Venne
Annie
A
Solomon
Scott R
SR
Fan
Yong
Y
Gostick
Emma
E
Price
David A
DA
Scotto
Christian
C
Read
Elizabeth J
EJ
Barrett
A John
AJ
D000938:Antigen-Presenting Cells / Q000187:drug effects / Q000276:immunology* / Q000528:radiation effects; D015704:CD4 Antigens / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects / Q000528:radiation effects; D005362:Ficoll; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006650:Histocompatibility Testing; D006801:Humans; D007109:Immunity / Q000187:drug effects / Q000528:radiation effects; D007376:Interleukin-2 / Q000494:pharmacology; D042541:Intracellular Space / Q000187:drug effects; D008027:Light; D008212:Lymphocyte Depletion / Q000379:methods*; D016853:Muromonab-CD3 / Q000494:pharmacology; D012235:Rhodamines / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000166:cytology* / Q000187:drug effects / Q000276:immunology* / Q000382:microbiology / Q000821:virology; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects / Q000276:immunology / Q000528:radiation effects; D013997:Time Factors; D014019:Tissue Donors
D015704:CD4 Antigens; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D007376:Interleukin-2; D016853:Muromonab-CD3; D012235:Rhodamines; D005362:Ficoll; C121365:4,5-dibromorhodamine 123