Generation of highly purified and functionally active human TH1 cells against Aspergillus fumigatus
Generation of highly purified and functionally active human TH1 cells against Aspergillus fumigatus
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DOI：
10.1182/blood-2005-04-1660
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发表时间：
2006-03-15
期刊：
影响因子：
20.3
通讯作者：
Lehrnbecher, T
中科院分区：
文献类型：
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作者：
Beck, O;Topp, MS;Lehrnbecher, T
侵袭性曲霉病仍然是接受同种异体干细胞移植（SCT）患者的严重并发症。由于淋巴细胞提供了针对真菌的关键次级防御，因此功能活跃的抗曲霉T细胞的过继转移可能是恢复适应性免疫效应机制的一种选择。利用干扰素(IFN)- γ分泌试验，我们在烟曲霉细胞提取物的刺激下分离出人类活化的T细胞。将该细胞群培养14天，我们从7名健康个体中的7人的单次100毫升抽血中平均获得1.1 X 10(7)个细胞。在接下来的14天内，这些细胞在曲霉抗原刺激下扩增到平均2.0 × 10(8) t -辅助性1 （THII）细胞分泌ifn - γ。对各种真菌抗原提取物进行测试，烟曲霉、黄曲霉、黑曲霉和青霉菌的抗原提取物激活后，产生ifn - γ的CD3(+)/CD4（+）细胞的比例相似，而交替孢霉和白色念珠菌的抗原提取物激活后，没有观察到显著的ifn - γ产生。此外，生成的T细胞能够诱导烟曲霉菌丝损伤，并显著增加人中性粒细胞诱导的菌丝损伤。与原始细胞群相比，生成的抗曲霉T细胞CD4(+) T细胞介导的同种异体反应性明显降低。总之，我们提出了一种简单可行的策略，可以从单次抽血中快速产生大量抗曲霉菌的功能性活性T细胞。我们的数据表明，抗曲霉的功能性活性T细胞可能是接受同种异体SCT患者的一种有希望的治疗选择。
Invasive aspergillosis remains a serious complication in patients undergoing allogeneic stem cell transplantation (SCT). Since it became clear that lymphocytes provide a critical secondary defense against fungi, adoptive transfer of functionally active anti-Aspergillus T cells might be an option to restore adaptive immune effector mechanisms. Using the interferon (IFN)-gamma secretion assay, we isolated human activated T cells upon stimulation with a cellular extract of Aspergillus fumigatus. Culturing this cell population for 14 days, we obtained an average of 1.1 X 10(7) cells from a single 100-mL blood draw in 7 of 7 healthy individuals. Within another 14 days, these cells were expanded to an average number of 2.0 X 10(8) T-helper 1 (THII) cells secreting IFN-gamma on stimulation with Aspergillus antigens. Testing various fungal antigen extracts, similar proportions of IFN-gamma-producing CD3(+)/CD4(+) cells were obtained upon activation with antigen extracts of A fumigatus, A flavus, A niger, and Penicillium chrysogenum, whereas no significant IFN-gamma production was observed upon activation with antigen extracts of Alternaria alternata and Candida albicans. In addition, generated T cells were able to induce damage to A fumigatus hyphae, and significantly increased hyphal damage induced by human neutrophils. CD4(+) T-cell-mediated alloreactivity of generated anti-Aspergillus T cells was clearly reduced compared with that of the original cell population. In conclusion, we present a simple and feasible strategy for rapid generation of a high number of functional active T cells against Aspergilius from a single blood draw. Our data suggest that functionally active T cells against Aspergillus could be a promising treatment option for patients undergoing allogeneic SCT.