Isolation and analysis of residual leucocytes from leucoreduced red blood cell units.

IF 1.8 4区 医学 Q3 HEMATOLOGY
Vox Sanguinis Pub Date : 2024-11-25 DOI:10.1111/vox.13775
Rena Hirani, Melinda M Dean, David O Irving
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引用次数: 0

Abstract

Background and objectives: Leucoreduction is used to remove donor leucocytes during red blood cell (RBC) manufacture. However, not all are removed, and long-term survival of donor leucocytes, termed transfusion-associated microchimerism (TAM), has been shown to occur in some patients following RBC transfusion. The mechanism of TAM occurrence is unknown. One hypothesis is that viable donor haematopoietic cells remain within RBC units that could engraft. However, the analysis of cells remaining within leucoreduced RBC units has been minimal. This study aimed to isolate and analyse any residual leucocytes recovered from leucoreduced RBC units.

Materials and methods: Leucoreduced RBC units were analysed on Day 1 (n = 4) and Day 42 (n = 4) post collection. Residual leucocytes were isolated using the EasySep™ RBC Depletion Reagent. Cell type analysis was conducted by flow cytometry using a leucocount reagent, a viability marker (7-amino-actinomycin D [7AAD]) and specific antibodies to CD45 and CD34. A representative 'pre-filter' sample was also obtained at the time of whole-blood donation to ensure expected cell counts across the donor samples.

Results: Analysis of the pre-filter sample showed that CD45+/CD34+ cells accounted for 0.02%-0.07% of all leucocytes. Up to 253,850 residual leucocytes were isolated across both storage timepoints, and of these, up to 48 cells were CD45+/CD34+/7AAD-.

Conclusion: Viable CD45+/CD34+ cells were isolated from leucoreduced RBC units, indicating the potential for donor progenitor cells to be present during transfusion. Further characterization of these residual cells is required to explain how TAM may occur in some patients following RBC transfusion.

白细胞还原红细胞单位中残留白细胞的分离与分析。
背景和目的:在红细胞(RBC)制造过程中,白细胞还原法用于去除供体白细胞。然而,并非所有的供体白细胞都被去除,在输注红细胞后,一些患者体内会出现供体白细胞的长期存活,即输血相关小嵌合体(TAM)。TAM 发生的机制尚不清楚。一种假设是,有活力的捐献者造血细胞残留在可移植的 RBC 单位中。然而,对残留在白细胞诱导的 RBC 单位中的细胞进行分析的研究却很少。本研究旨在分离和分析从去白细胞RBC单位中回收的任何残留白细胞:在采集后第 1 天(n = 4)和第 42 天(n = 4)对白细胞还原红细胞单位进行分析。使用 EasySep™ RBC 去白细胞试剂分离残留的白细胞。使用白细胞计数试剂、活力标记物(7-氨基放线菌素 D [7AAD])以及 CD45 和 CD34 的特异性抗体,通过流式细胞术进行细胞类型分析。在捐献全血时,还采集了一份具有代表性的 "预滤器 "样本,以确保所有捐献者样本中都有预期的细胞数量:对预过滤器样本的分析表明,CD45+/CD34+细胞占所有白细胞的0.02%-0.07%。在两个储存时间点分离出的残留白细胞多达 253850 个,其中 CD45+/CD34+/7AAD- 细胞多达 48 个:结论:从白细胞还原的红细胞单位中分离出了可存活的 CD45+/CD34+ 细胞,这表明在输血过程中可能存在供体祖细胞。需要进一步确定这些残留细胞的特征,以解释某些患者在输注 RBC 后为何会出现 TAM。
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来源期刊
Vox Sanguinis
Vox Sanguinis 医学-血液学
CiteScore
4.40
自引率
11.10%
发文量
156
审稿时长
6-12 weeks
期刊介绍: Vox Sanguinis reports on important, novel developments in transfusion medicine. Original papers, reviews and international fora are published on all aspects of blood transfusion and tissue transplantation, comprising five main sections: 1) Transfusion - Transmitted Disease and its Prevention: Identification and epidemiology of infectious agents transmissible by blood; Bacterial contamination of blood components; Donor recruitment and selection methods; Pathogen inactivation. 2) Blood Component Collection and Production: Blood collection methods and devices (including apheresis); Plasma fractionation techniques and plasma derivatives; Preparation of labile blood components; Inventory management; Hematopoietic progenitor cell collection and storage; Collection and storage of tissues; Quality management and good manufacturing practice; Automation and information technology. 3) Transfusion Medicine and New Therapies: Transfusion thresholds and audits; Haemovigilance; Clinical trials regarding appropriate haemotherapy; Non-infectious adverse affects of transfusion; Therapeutic apheresis; Support of transplant patients; Gene therapy and immunotherapy. 4) Immunohaematology and Immunogenetics: Autoimmunity in haematology; Alloimmunity of blood; Pre-transfusion testing; Immunodiagnostics; Immunobiology; Complement in immunohaematology; Blood typing reagents; Genetic markers of blood cells and serum proteins: polymorphisms and function; Genetic markers and disease; Parentage testing and forensic immunohaematology. 5) Cellular Therapy: Cell-based therapies; Stem cell sources; Stem cell processing and storage; Stem cell products; Stem cell plasticity; Regenerative medicine with cells; Cellular immunotherapy; Molecular therapy; Gene therapy.
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