Establishment of the Dual Humanized TK-NOG Mouse Model for HIV-associated Liver Pathogenesis
Establishment of the Dual Humanized TK-NOG Mouse Model for HIV-associated Liver Pathogenesis
复制标题
DOI：
10.3791/58645
复制
发表时间：
2019-09-01
影响因子：
1.2
通讯作者：
Poluektova, Larisa Y.
中科院分区：
文献类型：
--
作者：
Dagur, Raghubendra Singh;Wang, Weimin;Poluektova, Larisa Y.
尽管感染人类免疫缺陷病毒-1（HIV-1）的患者的预期寿命增加，但肝病已成为其发病的常见原因。由HIV-1引起的肝脏免疫病理学仍然难以捉摸。具有人类肝细胞和人类免疫系统的小型异种移植动物模型可以概括疾病发病机制的人类生物学。本文描述了通过人肝细胞和CD 34（+）造血干/祖细胞（HSPC）移植建立双重人源化小鼠模型的方案，以研究在HIV感染患者中观察到的肝脏免疫病理学。为了实现双重重建，向雄性TK-NOG（NOD.Cg-Prkdc（scid）Il 2 rg（tm 1 Sug）Tg（Alb-TK）7-2/ShiJic）小鼠腹膜内注射更昔洛韦（GCV）剂量以消除小鼠转基因肝细胞，并注射曲硫凡用于非清髓性调节，这两者都促进人肝细胞（HEP）移植和人免疫系统（HIS）发育。评估人白蛋白（ALB）水平用于肝移植，通过流式细胞术检测血液中人免疫细胞的存在证实了人免疫系统的建立。使用本文描述的方案开发的模型类似于HIV-1感染引起的肝损伤的多个组成部分。它的建立可能对于肝炎病毒合并感染的研究以及抗病毒和抗逆转录病毒药物的评估至关重要。
Despite the increased life expectancy of patients infected with human immunodeficiency virus-1 (HIV-1), liver disease has emerged as a common cause of their morbidity. The liver immunopathology caused by HIV-1 remains elusive. Small xenograft animal models with human hepatocytes and human immune system can recapitulate the human biology of the disease's pathogenesis. Herein, a protocol is described to establish a dual humanized mouse model through human hepatocytes and CD34(+) hematopoietic stem/progenitor cells (HSPCs) transplantation, to study liver immunopathology as observed in HIV-infected patients. To achieve dual reconstitution, male TK-NOG (NOD.Cg-Prkdc(scid) Il2rg(tm1Sug) Tg(Alb-TK)7-2/ShiJic) mice are intraperitoneally injected with ganciclovir (GCV) doses to eliminate mouse transgenic liver cells, and with treosulfan for nonmyeloablative conditioning, both of which facilitate human hepatocyte (HEP) engraftment and human immune system (HIS) development. Human albumin (ALB) levels are evaluated for liver engraftment, and the presence of human immune cells in blood detected by flow cytometry confirms the establishment of human immune system. The model developed using the protocol described here resembles multiple components of liver damage from HIV-1 infection. Its establishment could prove to be essential for studies of hepatitis virus co-infection and for the evaluation of antiviral and antiretroviral drugs.