D Nicole Worsham1, Jo-Anna Reems2, Zbigniew M Szczepiorkowski3, David H McKenna4, Thomas Leemhuis1, Aby J Mathew5, Jose A Cancelas1. 1. Hoxworth Blood Center, University of Cincinnati, Cincinnati, Ohio. 2. University of Utah School of Medicine, Salt Lake City, Utah. 3. Geisel School of Medicine at Dartmouth, Lebanon, New Hampshire. 4. University of Minnesota Medical School, Minneapolis, Minnesota. 5. BioLife Solutions, Inc., Bothell, Washington.
Abstract
BACKGROUND: Cryopreserved donor lymphocyte infusion (DLI) products are manufactured and administered to treat relapse after allogeneic hematopoietic stem cell transplantation. Reported clinical responses to DLIs vary broadly, even within the same group of patients. While there is an implicit recognition of the fact that different manufacturing protocols may have specific effects on different cell types, cryopreservation protocols are frequently derived from our experience in the cryopreservation of stem cell products and do not account for the heterogeneous functional nature of DLI T-cell populations. Here, we report the results of a prospective, multicenter trial on the effect of four different cryopreservation solutions that were used to freeze DLIs compared to control DLIs that were refrigerated overnight. STUDY DESIGN AND METHODS: Cryopreserved postthawed and refrigerated specimens were analyzed side by side for their T-cell subpopulation content and viability, as well as T-cell proliferation, cytokine secretion, and cytotoxic activities. RESULTS: This study indicates that "homemade" 10% dimethyl sulfoxide (DMSO) results in reduced viability of different CD4+ T-cell populations, including T-helper, T-cytotoxic, and T-regulatory populations, and a decrease in their proliferative and cytotoxic response to immunologically relevant stimuli, while the use of solutions containing 5% DMSO with intracellular-like cryoprotectant stabilizers maintains T-cell function at levels similar to refrigerated control samples. CONCLUSION: This study has important implications in determining the best cryoprotectant solution for specific clinical applications in allogeneic immunotherapy.
BACKGROUND: Cryopreserved donor lymphocyte infusion (DLI) products are manufactured and administered to treat relapse after allogeneic hematopoietic stem cell transplantation. Reported clinical responses to DLIs vary broadly, even within the same group of patients. While there is an implicit recognition of the fact that different manufacturing protocols may have specific effects on different cell types, cryopreservation protocols are frequently derived from our experience in the cryopreservation of stem cell products and do not account for the heterogeneous functional nature of DLI T-cell populations. Here, we report the results of a prospective, multicenter trial on the effect of four different cryopreservation solutions that were used to freeze DLIs compared to control DLIs that were refrigerated overnight. STUDY DESIGN AND METHODS: Cryopreserved postthawed and refrigerated specimens were analyzed side by side for their T-cell subpopulation content and viability, as well as T-cell proliferation, cytokine secretion, and cytotoxic activities. RESULTS: This study indicates that "homemade" 10% dimethyl sulfoxide (DMSO) results in reduced viability of different CD4+ T-cell populations, including T-helper, T-cytotoxic, and T-regulatory populations, and a decrease in their proliferative and cytotoxic response to immunologically relevant stimuli, while the use of solutions containing 5% DMSO with intracellular-like cryoprotectant stabilizers maintains T-cell function at levels similar to refrigerated control samples. CONCLUSION: This study has important implications in determining the best cryoprotectant solution for specific clinical applications in allogeneic immunotherapy.
Authors: Sandhya R Panch; Sandeep K Srivastava; Nasha Elavia; Andrew McManus; Shutong Liu; Ping Jin; Steven L Highfill; Xiaobai Li; Pradeep Dagur; James N Kochenderfer; Terry J Fry; Crystal L Mackall; Daniel Lee; Nirali N Shah; David F Stroncek Journal: Mol Ther Date: 2019-05-30 Impact factor: 11.454
Authors: Adam Joules; Julianne Connors; Rachel Johnson; Elizabeth A Van Orsow; David H McKenna; Sarah Nikiforow; Jerome Ritz; Adrian Gee; Allison Hubel Journal: Cytotherapy Date: 2021-01-30 Impact factor: 6.196