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Clinical Trial
. 2017 May;57(5):1133-1141.
doi: 10.1111/trf.14003. Epub 2017 Feb 24.

Autologous lymphapheresis for the production of chimeric antigen receptor T cells

Elizabeth S Allen  1 , David F Stroncek  1 , Jiaqiang Ren  1 , Anne F Eder  1 , Kamille A West  1 , Terry J Fry  2 , Daniel W Lee  3 , Crystal L Mackall  2 , Cathy Conry-Cantilena  1
Affiliations
Clinical Trial

Autologous lymphapheresis for the production of chimeric antigen receptor T cells

Elizabeth S Allen et al. Transfusion. 2017 May.

Abstract

Background: The first step in manufacturing chimeric antigen receptor (CAR) T cells is to collect autologous CD3+ lymphocytes by apheresis. Patients, however, often have leukopenia or have other disease-related complications. We evaluated the feasibility of collecting adequate numbers of CD3+ cells, risk factors for inadequate collections, and the rate of adverse events.

Study design and methods: Apheresis lymphocyte collections from patients participating in three CAR T-cell clinical trials were reviewed. Collections were performed on the COBE Spectra by experienced nurses, with the goal of obtaining a minimum of 0.6 × 109 and a target of 2 × 109 CD3+ cells. Preapheresis peripheral blood counts, apheresis parameters, and product cell counts were analyzed.

Results: Of the 71 collections, 69 (97%) achieved the minimum and 55 (77%) achieved the target. Before apheresis, the 16 patients with yields below the target had significantly lower proportions and absolute numbers of circulating lymphocytes and CD3+ lymphocytes and higher proportions of circulating blasts and NK cells than those who achieved the target (470 × 106 lymphocytes/L vs. 1340 × 106 lymphocytes/L, p = 0.008; 349 × 106 CD3+ cells/L vs. 914 × 106 CD3+ cells/L, p = 0.001; 17.6% blasts vs. 4.55% blasts, p = 0.029). Enrichment of blasts in the product compared to the peripheral blood occurred in four patients, including the two patients whose collections did not yield the minimum number of CD3+ cells. Apheresis complications occurred in 11 patients (15%) and, with one exception, were easily managed in the apheresis clinic.

Conclusions: In most patients undergoing CAR T-cell therapy, leukapheresis is well tolerated, and adequate numbers of CD3+ lymphocytes are collected.

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Figures

Figure 1
Figure 1. Apheresis CD3+ cell collection yield
Panel A. Total CD3+ cell yield by diagnosis. Each circle represents 1 patient. Panel B. CD3+ cell yield per liter of blood processed versus pre-apheresis CD3+ count. Using the pre-procedure CD3+ count (cells/μL) as the independent variable, x, the collection yield (CD3+ cells per L processed) can be estimated by the equation y=469394x + 108 (R=0.8613; R2=0.7419). Patients with acute leukemia are represented by closed diamonds, patients with solid tumors by open diamonds.
Figure 1
Figure 1. Apheresis CD3+ cell collection yield
Panel A. Total CD3+ cell yield by diagnosis. Each circle represents 1 patient. Panel B. CD3+ cell yield per liter of blood processed versus pre-apheresis CD3+ count. Using the pre-procedure CD3+ count (cells/μL) as the independent variable, x, the collection yield (CD3+ cells per L processed) can be estimated by the equation y=469394x + 108 (R=0.8613; R2=0.7419). Patients with acute leukemia are represented by closed diamonds, patients with solid tumors by open diamonds.
Figure 2
Figure 2. Comparison of absolute lymphocyte counts and peripheral blood NK cell (%) before apheresis in patients with below- and above-target CD3+ cell yields
Each circle represents 1 patient. The targeted yield was 2×109 CD3+ cells. Panel A. Absolute lymphocyte count in patients with below- and above-target CD3+ cell yields. A single patient with a yield above the target whose absolute lymphocyte count was 8.29×103/μL was not included. Panel B. Peripheral blood NK cells (%) in patients with below- and above-target CD3+ cell yields.
Figure 2
Figure 2. Comparison of absolute lymphocyte counts and peripheral blood NK cell (%) before apheresis in patients with below- and above-target CD3+ cell yields
Each circle represents 1 patient. The targeted yield was 2×109 CD3+ cells. Panel A. Absolute lymphocyte count in patients with below- and above-target CD3+ cell yields. A single patient with a yield above the target whose absolute lymphocyte count was 8.29×103/μL was not included. Panel B. Peripheral blood NK cells (%) in patients with below- and above-target CD3+ cell yields.
Figure 3
Figure 3. CD3+ cell collection yield vs total blood volumes processed
Each diamond represents 1 patient. Only the 16 patients whose collections yielded fewer than 2×109 CD3+ cells are depicted.
Figure 4
Figure 4. Difference in blast percentages in the apheresis product versus peripheral blood
Each circle represents 1 patient. Only those with circulating blasts on the day of collection are depicted.
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