Szabolcs Paul, Park Kyung-Duk, Reese Melissa, Marti Luciana, Broadwater Gloria, Kurtzberg Joanne
Department of Pediatrics, Pediatric Stem Cell Transplant Program, Duke University Medical Center, Durham, NC, USA.
Exp Hematol. 2003 Aug;31(8):708-14. doi: 10.1016/s0301-472x(03)00160-7.
Umbilical cord blood (UCB) T cells are predominantly CD45RA(+), secrete less cytokines, and have diminished cytotoxicity compared to adult peripheral blood (PB). We hypothesized that the functional impairment of bulk UCB cells results from the relative dominance of immature lymphocyte subsets. In this study we established the physiologic ranges of lymphocyte subsets in UCB, and contrasted those with adult PB.
Four-color FACS was utilized to characterize surface and intracellular protein expression on lymphocyte subsets from fresh unmanipulated UCB and adult PB.
We found that UCB contain significantly higher absolute numbers of T cells, NK cells, and B cells than adult PB (p<0.0001). UCB also contains more "naïve" cells not only among CD4(+) and CD8(+) T cells but also among B lymphocytes (p=0.003). Most UCB T cells are CD45RA(+)/CD62L(+) "recent thymic emigrants" with smaller TCRgammadelta (p<0.0001) and CD25(+) subsets (p=0.0068). Fewer UCB T cells display HLA-DR and CCR-5 activation markers (p<0.0001) while the CD8(+)/CD57(+)/CD28(-) "suppressor," CD8(+)/CD45RA(+)/CD27(-) "cytotoxic," and skin homing CLA(+) T-cell subsets are absent altogether. Compared with adult PB, more cord blood T cells progress through cell cycle (p<0.0001) and enter apoptosis (p=0.0003). Unlike in adult PB, the majority of proliferating Ki-67(+) T cells in UCB retain a CD45RA(+)/RO(-), CD69(-), CD25(-), HLA-DR(-) "resting" phenotype (p=0.0002).
Most T and B lymphocytes express a nai;ve phenotype in cord blood while "suppressor" and "cytotoxic" T-cell subsets are absent. Cycling UCB T cells retain a nai;ve immunophenotype that may represent homeostatic expansion rather than antigen-driven proliferation.
与成人外周血(PB)相比,脐带血(UCB)T细胞主要为CD45RA(+),分泌的细胞因子较少,细胞毒性也较低。我们推测,大量UCB细胞的功能受损是由于未成熟淋巴细胞亚群的相对优势所致。在本研究中,我们确定了UCB中淋巴细胞亚群的生理范围,并将其与成人PB进行了对比。
利用四色流式细胞术(FACS)对新鲜未处理的UCB和成人PB中淋巴细胞亚群的表面和细胞内蛋白表达进行表征。
我们发现,UCB中T细胞、NK细胞和B细胞的绝对数量显著高于成人PB(p<0.0001)。UCB中不仅CD4(+)和CD8(+) T细胞中,而且B淋巴细胞中也含有更多的“幼稚”细胞(p=0.003)。大多数UCB T细胞是CD45RA(+)/CD62L(+)“近期胸腺迁出者”,其TCRγδ较小(p<0.0001),CD25(+)亚群也较小(p=0.0068)。UCB T细胞中显示HLA-DR和CCR-5激活标志物的较少(p<0.0001),而CD8(+)/CD57(+)/CD28(-)“抑制性”、CD8(+)/CD45RA(+)/CD27(-)“细胞毒性”和皮肤归巢CLA(+) T细胞亚群则完全不存在。与成人PB相比,更多的脐带血T细胞进入细胞周期(p<0.0001)并进入凋亡(p=0.0003)。与成人PB不同,UCB中大多数增殖的Ki-67(+) T细胞保留CD45RA(+)/RO(-)、CD69(-)、CD25(-)、HLA-DR(-)“静止”表型(p=0.0002)。
脐带血中大多数T和B淋巴细胞表达幼稚表型,而“抑制性”和“细胞毒性”T细胞亚群不存在。循环的UCB T细胞保留幼稚免疫表型,这可能代表稳态扩增而非抗原驱动的增殖。