Conditional immortalization of human B cells by CD40 ligation.
Conditional immortalization of human B cells by CD40 ligation.
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DOI：
10.1371/journal.pone.0001464
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发表时间：
2008-01-23
期刊：
影响因子：
3.7
通讯作者：
Moosmann A
中科院分区：
文献类型：
--
作者：
Wiesner M;Zentz C;Mayr C;Wimmer R;Hammerschmidt W;Zeidler R;Moosmann A
一般认为，人类分化的细胞在体内具有有限的寿命和增殖能力，并且遗传修饰是其体外永生化的先决条件。在这里，我们重新讨论这个问题，研究人类B细胞的长期增殖潜力。早先显示，通过在白介素-4存在下刺激其受体CD 40，可以有效诱导来自健康供体外周血的人B细胞在体外增殖长达10周。当我们在改变细胞数量和培养物大小的条件下施加相同的刺激时，我们惊讶地发现，我们的处理诱导B细胞在目前长达1650天的观察期内增殖，代表超过370个群体倍增，这表明这些B细胞在体外是永生化的。可以从大多数健康成人供体中建立长期CD 40刺激的B细胞培养物。这些B细胞具有恒定的表型，不含EB病毒，并且仍然依赖于CD 40连接。它们具有组成性端粒酶活性和稳定的端粒长度。此外，它们对Toll样受体9配体的激活敏感，并且可用于体外扩增抗原特异性细胞毒性T细胞。我们的研究结果表明，人类体细胞可以逃避衰老，并有条件地永生化的外部刺激，而不需要遗传操作或肿瘤病毒感染。永生化人B细胞是免疫治疗和研究B细胞肿瘤发生、活化和功能的新工具。
It is generally assumed that human differentiated cells have a limited life-span and proliferation capacity in vivo, and that genetic modifications are a prerequisite for their immortalization in vitro. Here we readdress this issue, studying the long-term proliferation potential of human B cells. It was shown earlier that human B cells from peripheral blood of healthy donors can be efficiently induced to proliferate for up to ten weeks in vitro by stimulating their receptor CD40 in the presence of interleukin-4. When we applied the same stimuli under conditions of modified cell number and culture size, we were surprised to find that our treatment induced B cells to proliferate throughout an observation period of presently up to 1650 days, representing more than 370 population doublings, which suggested that these B cells were immortalized in vitro. Long-term CD40-stimulated B cell cultures could be established from most healthy adult human donors. These B cells had a constant phenotype, were free from Epstein-Barr virus, and remained dependent on CD40 ligation. They had constitutive telomerase activity and stabilized telomere length. Moreover, they were susceptible to activation by Toll-like receptor 9 ligands, and could be used to expand antigen-specific cytotoxic T cells in vitro. Our results indicate that human somatic cells can evade senescence and be conditionally immortalized by external stimulation only, without a requirement for genetic manipulation or oncoviral infection. Conditionally immortalized human B cells are a new tool for immunotherapy and studies of B cell oncogenesis, activation, and function.
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影响因子：
56.9
作者：
ARPIN, C;DECHANET, J;LIU, YJ
通讯作者：
LIU, YJ
影响因子：
56.9
作者：
BANCHEREAU, J;DEPAOLI, P;ROUSSET, F
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ROUSSET, F
影响因子：
4.4
作者：
Hooijberg, E;Ruizendaal, JJ;Spits, H
通讯作者：
Spits, H
影响因子：
5.4
作者：
Hammer, MH;Meyer, S;Reinke, P
通讯作者：
Reinke, P
影响因子：
15.3
作者：
Galibert, L;Burdin, N;deSaintVis, B;Garrone, P;VanKooten, C;Banchereau, J;Rousset, F
通讯作者：
Rousset, F