TY - JOUR
T1 - A New Method for Reactivating and Expanding T Cells Specific for Rhizopus oryzae
AU - Castillo, Paul
AU - Wright, Kaylor E.
AU - Kontoyiannis, Dimitrios P.
AU - Walsh, Thomas
AU - Patel, Shabnum
AU - Chorvinsky, Elizabeth
AU - Bose, Swaroop
AU - Hazrat, Yasmin
AU - Omer, Bilal
AU - Albert, Nathaniel
AU - Leen, Ann M.
AU - Rooney, Cliona M.
AU - Bollard, Catherine M.
AU - Cruz, Conrad Russell Y.
N1 - Publisher Copyright:
© 2018 The Authors
PY - 2018/6/15
Y1 - 2018/6/15
N2 - Mucormycosis is responsible for an increasing proportion of deaths after allogeneic bone marrow transplantation. Because this disease is associated with severe immunodeficiency and has shown resistance to even the newest antifungal agents, we determined the feasibility of reactivating and expanding Rhizopus oryzae-specific T cells for use as adoptive immunotherapy in transplant recipients. R. oryzae extract-pulsed monocytes were used to stimulate peripheral blood mononuclear cells from healthy donors, in the presence of different cytokine combinations. The generated R. oryzae-specific T cell products were phenotyped after the third stimulation and further characterized by the use of antibodies that block class I/II molecules, as well as pattern recognition receptors. Despite the very low frequency of R. oryzae-specific T cells of healthy donors, we found that stimulation with interleukin-2 (IL-2)/IL-7 cytokine combination could expand these rare cells. The expanded populations included 17%–83% CD4+ T cells that were specific for R. oryzae antigens. Besides interferon-γ (IFN-γ), these cells secreted IL-5, IL-10, IL-13, and tumor necrosis factor alpha (TNF-α), and recognized fungal antigens presented by HLA-II molecules rather than through nonspecific signaling. The method described herein is robust and reproducible, and could be used to generate adequate quantities of activated R. oryzae-specific T cells for clinical testing of safety and antifungal efficacy in patients with mucormycosis.
AB - Mucormycosis is responsible for an increasing proportion of deaths after allogeneic bone marrow transplantation. Because this disease is associated with severe immunodeficiency and has shown resistance to even the newest antifungal agents, we determined the feasibility of reactivating and expanding Rhizopus oryzae-specific T cells for use as adoptive immunotherapy in transplant recipients. R. oryzae extract-pulsed monocytes were used to stimulate peripheral blood mononuclear cells from healthy donors, in the presence of different cytokine combinations. The generated R. oryzae-specific T cell products were phenotyped after the third stimulation and further characterized by the use of antibodies that block class I/II molecules, as well as pattern recognition receptors. Despite the very low frequency of R. oryzae-specific T cells of healthy donors, we found that stimulation with interleukin-2 (IL-2)/IL-7 cytokine combination could expand these rare cells. The expanded populations included 17%–83% CD4+ T cells that were specific for R. oryzae antigens. Besides interferon-γ (IFN-γ), these cells secreted IL-5, IL-10, IL-13, and tumor necrosis factor alpha (TNF-α), and recognized fungal antigens presented by HLA-II molecules rather than through nonspecific signaling. The method described herein is robust and reproducible, and could be used to generate adequate quantities of activated R. oryzae-specific T cells for clinical testing of safety and antifungal efficacy in patients with mucormycosis.
KW - antigen presentation/processing
KW - cytokines
KW - cytotoxic T cells
KW - fungal
KW - immune reconstitution
KW - transplantation
UR - http://www.scopus.com/inward/record.url?scp=85054019803&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85054019803&partnerID=8YFLogxK
U2 - 10.1016/j.omtm.2018.03.003
DO - 10.1016/j.omtm.2018.03.003
M3 - Article
C2 - 30038934
AN - SCOPUS:85054019803
SN - 2329-0501
VL - 9
SP - 305
EP - 312
JO - Molecular Therapy - Methods and Clinical Development
JF - Molecular Therapy - Methods and Clinical Development
ER -