体外扩增的脐带血T细胞维持幼稚表型和TCR多样性。
Ex vivo expanded umbilical cord blood T cells maintain naive phenotype and TCR diversity.
作者信息
Parmar S, Robinson S N, Komanduri K, St John L, Decker W, Xing D, Yang H, McMannis J, Champlin R, de Lima M, Molldrem J, Rieber A, Bonyhadi M, Berenson R, Shpall E J
机构信息
The Department of Blood and Marrow Transplantation, MD Anderson Cancer Center, Houston, Texas, USA.
出版信息
Cytotherapy. 2006;8(2):149-57. doi: 10.1080/14653240600620812.
BACKGROUND
Umbilical cord blood (CB) is a promising source of hematopoietic stem cells for allogeneic transplantation. However, delayed engraftment and impaired immune reconstitution remain major limitations. Enrichment of donor grafts with CB T cells expanded ex vivo might facilitate improved T-cell immune reconstitution post-transplant. We hypothesized that CB T cells could be expanded using paramagnetic microbeads covalently linked to anti-CD3 and anti-CD28 Ab.
METHODS
CB units were divided into three fractions: (1) cells cultured without beads, (2) cells cultured with beads and (3) cells cultured with beads following CD3+ magnetic enrichment. All fractions were cultured for 14 days in the presence of IL-2 (200 IU/mL).
RESULTS
A mean 100-fold expansion (range 49-154) of total nucleated cells was observed in the CD3+ magnetically enriched fraction. Following expansion, CB T cells retained a naive and/or central memory phenotype and contained a polyclonal TCR diversity demonstrated by spectratyping.
DISCUSSION
Our data provide evidence that naive and diverse CB T cells may be expanded ex vivo and warrant additional studies in the setting of human CB transplantation.
背景
脐带血（CB）是同种异体移植中造血干细胞的一个有前景的来源。然而，移植延迟和免疫重建受损仍然是主要限制因素。用体外扩增的CB T细胞富集供体移植物可能有助于改善移植后T细胞免疫重建。我们假设可以使用与抗CD3和抗CD28抗体共价连接的顺磁性微珠来扩增CB T细胞。
方法
将CB样本分为三个部分：（1）无微珠培养的细胞，（2）有微珠培养的细胞，（3）CD3 + 磁性富集后有微珠培养的细胞。所有部分在IL-2（200 IU/mL）存在下培养14天。
结果
在CD3 + 磁性富集部分观察到有核细胞平均扩增100倍（范围49 - 154）。扩增后，CB T细胞保留了幼稚和/或中央记忆表型，并通过光谱分型显示含有多克隆TCR多样性。
讨论
我们的数据提供了证据，表明幼稚且多样的CB T细胞可以在体外扩增，并且值得在人类CB移植背景下进行进一步研究。
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