Automated Cell Enrichment of Cytomegalovirus-specific T cells for Clinical Applications using the Cytokine-capture System.
The adoptive transfer of pathogen-specific T cells can be used to prevent and treat opportunistic infections such as cytomegalovirus (CMV) infection occurring after allogeneic hematopoietic stem-cell transplantation. Viral-specific T cells from allogeneic donors, including third party donors, can be propagated ex vivo in compliance with current good manufacturing practice (cGMP), employing repeated rounds of antigen-driven stimulation to selectively propagate desired T cells. The identification and isolation of antigen-specific T cells can also be undertaken based upon the cytokine capture system of T cells that have been activated to secrete gamma-interferon (IFN-γ). However, widespread human application of the cytokine capture system (CCS) to help restore immunity has been limited as the production process is time-consuming and requires a skilled operator. The development of a second-generation cell enrichment device such as CliniMACS Prodigy now enables investigators to generate viral-specific T cells using an automated, less labor-intensive system. This device separates magnetically labeled cells from unlabeled cells using magnetic activated cell sorting technology to generate clinical-grade products, is engineered as a closed system and can be accessed and operated on the benchtop. We demonstrate the operation of this new automated cell enrichment device to manufacture CMV pp65-specific T cells obtained from a steady-state apheresis product obtained from a CMV seropositive donor. These isolated T cells can then be directly infused into a patient under institutional and federal regulatory supervision. All the bio-processing steps including removal of red blood cells, stimulation of T cells, separation of antigen-specific T cells, purification, and washing are fully automated. Devices such as this raise the possibility that T cells for human application can be manufactured outside of dedicated good manufacturing practice (GMP) facilities and instead be produced in blood banking facilities where staff can supervise automated protocols to produce multiple products.
Division of Pediatrics, U.T. MD Anderson Cancer Center.
Pappanaicken
P
Kumaresan
Division of Pediatrics, U.T. MD Anderson Cancer Center.
Mathew
M
Figliola
Division of Pediatrics, U.T. MD Anderson Cancer Center.
Judy S
JS
Moyes
Division of Pediatrics, U.T. MD Anderson Cancer Center.
M Helen
MH
Huls
Division of Pediatrics, U.T. MD Anderson Cancer Center.
Priti
P
Tewari
Stem Cell Transplantation and Cellular Therapy, U.T. MD Anderson Cancer Center.
Elizabeth J
EJ
Shpall
Stem Cell Transplantation and Cellular Therapy, U.T. MD Anderson Cancer Center.
Richard
R
Champlin
Division of Pediatrics, U.T. MD Anderson Cancer Center; ljncooper@mdanderson.org.
Laurence J N
LJ
Cooper
D001781:Blood Component Removal / Q000379:methods*; D003587:Cytomegalovirus / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000276:immunology / Q000821:virology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D010750:Phosphoproteins / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000821:virology*; D014763:Viral Matrix Proteins / Q000276:immunology
D018984:Epitopes, T-Lymphocyte; D010750:Phosphoproteins; D014763:Viral Matrix Proteins; C065418:cytomegalovirus matrix protein 65kDa