ORIGINAL RESEARCH article
Front. Med.
Sec. Translational Medicine
Recovery Efficiency and Functional Characterization of T cells and NK Cells from Leukocyte Filters
Provisionally accepted
- 1Shenzhen Blood Center, Shenzhen, China
- 2The Eighth Affiliated Hospital of Sun Yat-Sen University, Shenzhen, China
- 3The Third People’s Hospital of Longgang, Shenzhen, China
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Objective: This study aimed to investigate the recovery efficiency and functional characteristics of lymphocytes isolated from leukocyte filters, compared with those isolated directly from whole blood sample, and to assess their potential as an alternative immune cell source for therapeutic applications. Methods: From December 2023 to 2024, leukocyte filters and 5 mL whole blood samples were collected from healthy donors. Cell recovery was optimized by testing different flushing methods (forward/reverse), solutions (1×PBS / NS), and volumes (20 – 80 mL). Peripheral blood mononuclear cells (PBMCs) were isolated via density gradient centrifugation, and lymphocyte subsets (CD3+CD56+ NK, CD3+CD4+ T, CD3+CD8+ T cells) were analyzed by flow cytometry. Transcriptome sequencing was conducted to evaluate functional differences. Results: Reverse flushing with 40 mL normal saline (NS) achieved cost-effective lymphocyte recovery, with cell counts correlating with flushing method and volume. The proportion of CD3+CD4+ T cells in 40 mL NS was higher than that in whole blood samples (P < 0.05), while CD3+CD8+ T cells was lower (P < 0.01). CD3+CD4+ T cells exhibited the highest recovery efficiency, and CD3+CD8+ T cells the lowest. Transcriptome analysis identified 634 differentially expressed genes, however, GO/KEGG enrichment analysis revealed no significant pathways related to NK or T cell functions (P-adjust > 0.05). Conclusion: Reverse flushing of leukocyte filters with 40 mL NS enables cost-effective recovery of lymphocytes, particularly CD3+CD4+ T cells. The recovered NK and T cells exhibit functional equivalence to those from those from whole blood samples, offering a low-cost and sustainable cell source for allogeneic cell therapy.
Keywords: Leukocyte filter, whole blood, NK cells, T cells, Transcriptome sequencing
Received: 07 Aug 2025; Accepted: 17 Nov 2025.
Copyright: © 2025 Sun, Li, Cai, Wang and Zeng. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Songxing Wang, wsxdxyx2003@163.com
Jinfeng Zeng, zzengjf@163.com
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