Shultz Leonard D, Lyons Bonnie L, Burzenski Lisa M, Gott Bruce, Chen Xiaohua, Chaleff Stanley, Kotb Malak, Gillies Stephen D, King Marie, Mangada Julie, Greiner Dale L, Handgretinger Rupert
The Jackson Laboratory, Bar Harbor, ME 04609, USA.
J Immunol. 2005 May 15;174(10):6477-89. doi: 10.4049/jimmunol.174.10.6477.
Ethical considerations constrain the in vivo study of human hemopoietic stem cells (HSC). To overcome this limitation, small animal models of human HSC engraftment have been used. We report the development and characterization of a new genetic stock of IL-2R common gamma-chain deficient NOD/LtSz-scid (NOD-scid IL2Rgamma(null)) mice and document their ability to support human mobilized blood HSC engraftment and multilineage differentiation. NOD-scid IL2Rgamma(null) mice are deficient in mature lymphocytes and NK cells, survive beyond 16 mo of age, and even after sublethal irradiation resist lymphoma development. Engraftment of NOD-scid IL2Rgamma(null) mice with human HSC generate 6-fold higher percentages of human CD45(+) cells in host bone marrow than with similarly treated NOD-scid mice. These human cells include B cells, NK cells, myeloid cells, plasmacytoid dendritic cells, and HSC. Spleens from engrafted NOD-scid IL2Rgamma(null) mice contain human Ig(+) B cells and lower numbers of human CD3(+) T cells. Coadministration of human Fc-IL7 fusion protein results in high percentages of human CD4(+)CD8(+) thymocytes as well human CD4(+)CD8(-) and CD4(-)CD8(+) peripheral blood and splenic T cells. De novo human T cell development in NOD-scid IL2Rgamma(null) mice was validated by 1) high levels of TCR excision circles, 2) complex TCRbeta repertoire diversity, and 3) proliferative responses to PHA and streptococcal superantigen, streptococcal pyrogenic exotoxin. Thus, NOD-scid IL2Rgamma(null) mice engrafted with human mobilized blood stem cells provide a new in vivo long-lived model of robust multilineage human HSC engraftment.
伦理考量限制了人类造血干细胞(HSC)的体内研究。为克服这一限制,已采用人类HSC植入的小动物模型。我们报告了一种新的白细胞介素2受体共同γ链缺陷型NOD/LtSz-scid(NOD-scid IL2Rγ(null))小鼠遗传品系的开发与特性,并证明了它们支持人类动员的血液HSC植入和多谱系分化的能力。NOD-scid IL2Rγ(null)小鼠缺乏成熟淋巴细胞和自然杀伤(NK)细胞,存活时间超过16个月,即使在亚致死剂量照射后也能抵抗淋巴瘤的发生。用人类HSC植入NOD-scid IL2Rγ(null)小鼠后,宿主骨髓中人类CD45(+)细胞的百分比比同样处理的NOD-scid小鼠高6倍。这些人类细胞包括B细胞、NK细胞、髓样细胞、浆细胞样树突状细胞和HSC。植入NOD-scid IL2Rγ(null)小鼠的脾脏含有人类Ig(+) B细胞和数量较少的人类CD3(+) T细胞。同时给予人类Fc-IL7融合蛋白会导致人类CD4(+)CD8(+)胸腺细胞以及人类CD4(+)CD8(-)和CD4(-)CD8(+)外周血和脾脏T细胞的高百分比。通过以下几点验证了NOD-scid IL2Rγ(null)小鼠中人类T细胞的从头发育:1)高水平的T细胞受体切除环;2)复杂的TCRβ库多样性;3)对PHA和链球菌超抗原、链球菌致热外毒素的增殖反应。因此,植入人类动员的血液干细胞的NOD-scid IL2Rγ(null)小鼠提供了一种新的体内长期存在的强大多谱系人类HSC植入模型。