A novel predictive algorithm to personalize autologous T-cell harvest for chimeric antigen receptor T-cell manufacture

Published:December 11, 2022DOI:


      Background aims

      The most widely accepted starting materials for chimeric antigen receptor T-cell manufacture are autologous CD3+ T cells obtained via the process of leukapheresis, also known as T-cell harvest. As this treatment modality gains momentum and apheresis units struggle to meet demand for harvest slots, strategies to streamline this critical step are warranted.


      This retrospective review of 262 T-cell harvests, with a control cohort of healthy donors, analyzed the parameters impacting CD3+ T-cell yield in adults with B-cell malignancies. The overall aim was to design a novel predictive algorithm to guide the required processed blood volume (PBV) (L) on the apheresis machine to achieve a specific CD3+ target yield.


      Factors associated with CD3+ T-cell yield on multivariate analysis included peripheral blood CD3+ count (natural log, ×109/L), hematocrit (HCT) and PBV with coefficients of 0.86 (95% confidence interval [CI], 0.80–0.92, P < 0.001), 1.30 (95% CI, 0.51–2.08, P = 0.001) and 0.09 (95% CI, 0.07–0.11, P < 0.001), respectively. The authors’ model, incorporating CD3+ cell count, HCT and PBV (L), with an adjusted R2 of 0.87 and root-mean-square error of 0.26 in the training dataset, was highly predictive of CD3+ cell yield in the testing dataset. An online application to estimate PBV using this algorithm can be accessed at


      The authors propose a transferrable model that incorporates clinical and laboratory variables accessible pre-harvest for use across the field of T-cell therapy. Pending further validation, such a model may be used to generate an individual leukapheresis plan and streamline the process of cell harvest, a well-recognized bottleneck in the industry.

      Key Words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Cytotherapy
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Sadelain M
        • Brentjens R
        • Rivière I.
        The basic principles of chimeric antigen receptor design.
        Cancer discovery. 2013; 3: 388-398
        • Maude SL
        • Laetsch TW
        • Buechner J
        • et al.
        Tisagenlecleucel in children and young adults with B-cell lymphoblastic leukemia.
        New England Journal of Medicine. 2018; 378: 439-448
        • Schuster SJ
        • Bishop MR
        • Tam CS
        • et al.
        Tisagenlecleucel in adult relapsed or refractory diffuse large B-cell lymphoma.
        New England Journal of Medicine. 2019; 380: 45-56
        • Neelapu SS
        • Locke FL
        • Bartlett NL
        • et al.
        Axicabtagene ciloleucel CAR T-cell therapy in refractory large B-cell lymphoma.
        New England Journal of Medicine. 2017; 377: 2531-2544
        • Howell C
        • Douglas K
        • Cho G
        • et al.
        Guideline on the clinical use of apheresis procedures for the treatment of patients and collection of cellular therapy products.
        Transfusion Medicine. 2015; 25: 57-78
        • Yuan S
        • Ziman A
        • Smeltzer B
        • Lu Q
        • Goldfinger D.
        Moderate and severe adverse events associated with apheresis donations: incidences and risk factors.
        Transfusion. 2010; 50: 478-486
        • McGuirk J
        • Waller EK
        • Qayed M
        • et al.
        Building blocks for institutional preparation of CTL019 delivery.
        Cytotherapy. 2017; 19: 1015-1024
        • Hunt EA
        • Jain NG
        • Somers MJ.
        Apheresis therapy in children: an overview of key technical aspects and a review of experience in pediatric renal disease.
        Journal of clinical apheresis. 2013; 28: 36-47
        • Allen ES
        • Stroncek DF
        • Ren J
        • et al.
        Autologous lymphapheresis for the production of chimeric antigen receptor T cells.
        Transfusion. 2017; 57: 1133-1141
        • Ceppi F
        • Rivers J
        • Annesley C
        • et al.
        Lymphocyte apheresis for chimeric antigen receptor T-cell manufacturing in children and young adults with leukemia and neuroblastoma.
        Transfusion. 2018; 58: 1414-1420
        • Tuazon SA
        • Li A
        • Gooley T
        • et al.
        Factors affecting lymphocyte collection efficiency for the manufacture of chimeric antigen receptor T cells in adults with B-cell malignancies.
        Transfusion. 2019; 59: 1773-1780
        • Punzel M
        • Kozlova A
        • Quade A
        • Schmidt A
        • Smith R.
        Evolution of MNC and lymphocyte collection settings employing different Spectra Optia® leukapheresis systems.
        Vox sanguinis. 2017; 112: 586-594
        • Hutt D
        • Bielorai B
        • Baturov B
        • et al.
        Feasibility of leukapheresis for CAR T-cell production in heavily pre-treated pediatric patients.
        Transfusion and Apheresis Science. 2020; 59102769
        • Jarisch A
        • Rettinger E
        • Sörensen J
        • et al.
        Unstimulated apheresis for chimeric antigen receptor manufacturing in pediatric/adolescent acute lymphoblastic leukemia patients.
        Journal of Clinical Apheresis. 2020; 35: 398-405
        • Korell F
        • Laier S
        • Sauer S
        • et al.
        Current Challenges in Providing Good Leukapheresis Products for Manufacturing of CAR-T Cells for Patients with Relapsed/Refractory NHL or ALL.
        Cells. 2020; 9: 1225
        • Bersenev A
        • Kili S.
        Management of “out of specification” commercial autologous CAR-T cell products.
        Cell and Gene Therapy Insights. 2018; 4: 1051-1058
        • Carnoy S
        • Beaumont JL
        • Kanouni T
        • et al.
        How to perform leukapheresis for procurement of the staring material used for commercial CAR T-cell manufacturing: a consensus from experts convened by the SFGM-TC.
        Bulletin du cancer. 2021; 108: 295-303
        • Kansagra AJ
        • Frey NV
        • Bar M
        • et al.
        Clinical utilization of chimeric antigen receptor T cells in B cell acute Lymphoblastic Leukemia: An expert opinion from the European society for blood and marrow transplantation and the American society for transplantation and cellular therapy.
        Biology of Blood and Marrow Transplantation. 2019; 25: e76-e85
        • Hayden P
        • Roddie C
        • Bader P
        • et al.
        Management of Adults and Children receiving CAR T-cell therapy: 2021 Best Practice Recommendations of the European Society for Blood and Marrow Transplantation (EBMT) and the Joint Accreditation Committee of ISCT and EBMT (JACIE) and the European Haematology Association (EHA).
        Annals of Oncology. 2022; 33: 259-275
        • Yakoub-Agha I
        • Chabannon C
        • Bader P
        • et al.
        Management of adults and children undergoing chimeric antigen receptor T-cell therapy: best practice recommendations of the European Society for Blood and Marrow Transplantation (EBMT) and the Joint Accreditation Committee of ISCT and EBMT (JACIE).
        Haematologica. 2020; 105: 297-316
        • Neyrinck MM
        • Vrielink H
        Joint Task Force for Education and Certification. Calculations in apheresis.
        Journal of clinical apheresis. 2015; 30: 38-42
      1. Terumo Corporation. Spectra Optia Apheresis System Operator's Manual. (2018). Terumo BCT: Lakewood, Colorado. URL: Accessed 29 November 2022.

        • Harrell Jr, FE
        • Lee KL
        • Califf RM
        • Pryor DB
        • Rosati RA
        Regression modelling strategies for improved prognostic prediction.
        Statistics in medicine. 1984; 3: 143-152
        • Greenland S
        • Daniel R
        • Pearce N.
        Outcome modelling strategies in epidemiology: traditional methods and basic alternatives.
        International journal of epidemiology. 2016; 45: 565-575
        • White H.
        A heteroskedasticity-consistent covariance matrix estimator and a direct test for heteroskedasticity.
        Econometrica: journal of the Econometric Society. 1980; 48: 817-838
        • Collins GS
        • Reitsma JB
        • Altman DG
        • Moons KG.
        Transparent reporting of a multivariable prediction model for individual prognosis or diagnosis (TRIPOD): the TRIPOD statement.
        Journal of British Surgery. 2015; 102: 148-158
        • Chen J
        • Goss C
        • Avecilla ST
        • et al.
        Evaluation of peripheral blood mononuclear cell collection by leukapheresis.
        Transfusion. 2019; 59: 1765-1772
        • Anyanwu A
        • Sitzmann N
        • Hetjens S
        • et al.
        Low-Volume Leukapheresis in Non-Cytokine-Stimulated Donors for the Collection of Mononuclear Cells.
        Transfusion Medicine and Hemotherapy. 2018; 45: 323-330
        • Fesnak A
        • Lin C
        • Siegel DL
        • Maus MV.
        CAR-T cell therapies from the transfusion medicine perspective.
        Transfusion medicine reviews. 2016; 30: 139-145
        • Hupperetz C
        • Lah S
        • Kim H
        • Kim CH.
        CAR T Cell Immunotherapy beyond haematological malignancy.
        Immune Network. 2022; 22e6
        • Strasser EF
        • Zimmermann R
        • Weisbach V
        • Ringwald J
        • Zingsem J
        • Eckstein R.
        Mononuclear cell variability and recruitment in non-cytokine-stimulated donors after serial 10-liter leukapheresis procedures.
        Transfusion. 2005; 45: 445-452
        • Thibodeaux SR
        • Aqui NA
        • Park YA
        • et al.
        Lack of defined apheresis collection criteria in publicly available CAR-T cell clinical trial descriptions: comprehensive review of over 600 studies.
        Journal of Clinical Apheresis. 2022; 37: 223-236
        • Constantinou VC
        • Bouinta A
        • Karponi G
        • et al.
        Poor stem cell harvest may not always be related to poor mobilization: lessons gained from a mobilization study in patients with β-thalassemia major.
        Transfusion. 2017; 57: 1031-1039
        • Quinn KM
        • Fox A
        • Harland KL
        • et al.
        Age-related decline in primary CD8+ T cell responses is associated with the development of senescence in virtual memory CD8+ T cells.
        Cell reports. 2018; 23: 3512-3524
        • Schietinger A
        • Philip M
        • Krisnawan VE
        • et al.
        Tumor-specific T cell dysfunction is a dynamic antigen-driven differentiation program initiated early during tumorigenesis.
        Immunity. 2016; 45: 389-401
        • Das RK
        • Vernau L
        • Grupp SA
        • Barrett DM.
        Naive T-cell deficits at diagnosis and after chemotherapy impair cell therapy potential in pediatric cancers.
        Cancer discovery. 2019; 9: 492-499
        • Das RK
        • O'Connor RS
        • Grupp SA
        • Barrett DM
        Lingering effects of chemotherapy on mature T cells impair proliferation.
        Blood advances. 2020; 4: 4653-4664
        • Ghassemi S
        • Durgin JS
        • Nunez-Cruz S
        • et al.
        Rapid manufacturing of non-activated potent CAR T cells.
        Nature biomedical engineering. 2022; 6: 118-128
        • Ghassemi S
        • Nunez-Cruz S
        • O'Connor RS
        • et al.
        Reducing Ex Vivo Culture Improves the Antileukemic Activity of Chimeric Antigen Receptor (CAR) T CellsLimited Ex Vivo Culture Improves CAR T-cell Immunotherapy.
        Cancer immunology research. 2018; 6: 1100-1109
        • Yang J
        • He J
        • Zhang X
        • et al.
        A feasibility and safety study of a new CD19-directed fast CAR-T therapy for refractory and relapsed B cell acute lymphoblastic leukemia.
        Blood. 2019; 134: 825
        • Deng Q
        • Han G
        • Puebla-Osorio N
        • et al.
        Characteristics of anti-CD19 CAR T cell infusion products associated with efficacy and toxicity in patients with large B cell lymphomas.
        Nature medicine. 2020; 26: 1878-1887
        • Fraietta JA
        • Lacey SF
        • Orlando EJ
        • et al.
        Determinants of response and resistance to CD19 chimeric antigen receptor (CAR) T cell therapy of chronic lymphocytic leukemia.
        Nature medicine. 2018; 24: 563-571