PDF(Pubmed)
Abstract：
UNASSIGNED: Humanized mouse models to recapitulate human biological systems still have limitations, such as the onset of lethal graft-versus-host disease (GvHD), a variable success rate, and the low accessibility of total body irradiation (TBI). Recently, mice modified with the CD47-SIRPA axis have been studied to improve humanized mouse models. However, such trials have been rarely applied in NOD mice. In this study, we created a novel mouse strain, NOD-CD47nullRag2nullIL-2rγnull (RTKO) mice, and applied it to generate humanized mice.
UNASSIGNED: Four-week-old female NOD-Rag2nullIL-2rγnull (RID) and RTKO mice pre-conditioned with TBI or busulfan (BSF) injection were used for generating human CD34+ hematopoietic stem cell (HSC) engrafted humanized mice. Clinical signs were observed twice a week, and body weight was measured once a week. Flow cytometry for human leukocyte antigens was performed at intervals of four weeks or two weeks, and mice were sacrificed at 48 weeks after HSC injection.
UNASSIGNED: For a long period from 16 to 40 weeks post transplantation, the percentage of hCD45 was mostly maintained above 25% in all groups, and it was sustained the longest and highest in the RTKO BSF group. Reconstruction of human leukocytes, including hCD3, was also most prominent in the RTKO BSF group. Only two mice died before 40 weeks post transplantation in all groups, and there were no life-threatening GvHD lesions except in the dead mice. The occurrence of GvHD has been identified as mainly due to human T cells infiltrating tissues and their related cytokines.
UNASSIGNED: Humanized mouse models under all conditions applied in this study are considered suitable models for long-term experiments based on the improvement of human leukocytes reconstruction and the stable animal health. Especially, RTKO mice pretreated with BSF are expected to be a valuable platform not only for generating humanized mice but also for various immune research fields.
UNASSIGNED: Four-week-old female NOD-Rag2nullIL-2rγnull (RID) and RTKO mice pre-conditioned with TBI or busulfan (BSF) injection were used for generating human CD34+ hematopoietic stem cell (HSC) engrafted humanized mice. Clinical signs were observed twice a week, and body weight was measured once a week. Flow cytometry for human leukocyte antigens was performed at intervals of four weeks or two weeks, and mice were sacrificed at 48 weeks after HSC injection.
UNASSIGNED: For a long period from 16 to 40 weeks post transplantation, the percentage of hCD45 was mostly maintained above 25% in all groups, and it was sustained the longest and highest in the RTKO BSF group. Reconstruction of human leukocytes, including hCD3, was also most prominent in the RTKO BSF group. Only two mice died before 40 weeks post transplantation in all groups, and there were no life-threatening GvHD lesions except in the dead mice. The occurrence of GvHD has been identified as mainly due to human T cells infiltrating tissues and their related cytokines.
UNASSIGNED: Humanized mouse models under all conditions applied in this study are considered suitable models for long-term experiments based on the improvement of human leukocytes reconstruction and the stable animal health. Especially, RTKO mice pretreated with BSF are expected to be a valuable platform not only for generating humanized mice but also for various immune research fields.
摘要：
■用于概括人类生物系统的人性化小鼠模型仍然存在局限性，例如致命的移植物抗宿主病（GvHD）的发作，可变的成功率，以及全身照射(TBI)的低可及性。最近,已经研究了用CD47-SIRPA轴修饰的小鼠以改善人源化小鼠模型。然而,这种试验很少应用于NOD小鼠。在这项研究中,我们创造了一种新的老鼠品系，NOD-CD47nullRag2nullIL-2Rγnull（RTKO）小鼠，并将其应用于产生人源化小鼠。
■用TBI或白消安（BSF）注射预处理的四周龄雌性NOD-Rag2nullIL-2Rγnull（RID）和RTKO小鼠用于产生人CD34造血干细胞（HSC）移植的人源化小鼠。每周观察两次临床体征，每周测量一次体重。以4周或2周的间隔进行人白细胞抗原的流式细胞术。在HSC注射后48周处死小鼠。
■移植后16至40周，hCD45的百分比在所有组中大多保持在25%以上，在RTKOBSF组中持续时间最长，最高。人白细胞的重建，包括hCD3在内，在RTKOBSF组中也最为突出。在所有组中，只有两只小鼠在移植后40周前死亡，除了死亡的小鼠，没有危及生命的GvHD病变。GvHD的发生已被鉴定为主要归因于人T细胞浸润组织及其相关细胞因子。
■在本研究中应用的所有条件下的人源化小鼠模型被认为是基于人类白细胞重建的改善和稳定的动物健康的长期实验的合适模型。尤其是,用BSF预处理的RTKO小鼠有望成为不仅用于产生人源化小鼠而且用于各种免疫研究领域的有价值的平台。
■用TBI或白消安（BSF）注射预处理的四周龄雌性NOD-Rag2nullIL-2Rγnull（RID）和RTKO小鼠用于产生人CD34造血干细胞（HSC）移植的人源化小鼠。每周观察两次临床体征，每周测量一次体重。以4周或2周的间隔进行人白细胞抗原的流式细胞术。在HSC注射后48周处死小鼠。
■移植后16至40周，hCD45的百分比在所有组中大多保持在25%以上，在RTKOBSF组中持续时间最长，最高。人白细胞的重建，包括hCD3在内，在RTKOBSF组中也最为突出。在所有组中，只有两只小鼠在移植后40周前死亡，除了死亡的小鼠，没有危及生命的GvHD病变。GvHD的发生已被鉴定为主要归因于人T细胞浸润组织及其相关细胞因子。
■在本研究中应用的所有条件下的人源化小鼠模型被认为是基于人类白细胞重建的改善和稳定的动物健康的长期实验的合适模型。尤其是,用BSF预处理的RTKO小鼠有望成为不仅用于产生人源化小鼠而且用于各种免疫研究领域的有价值的平台。
