Mouse cell L1210-based HIV-1 infectible host cell

A technology for host cells and HIV-1, applied in the field of HIV-1 infectable host cells and their preparation, and can solve the problems of complete replication of difficult-to-maintain viruses and the like

Inactive Publication Date: 2017-07-11
深圳市龙华新区中心医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, HIV-1 host cells of non-human origin, especially cells of small animal origin, have been largely hindered in progress because of the species-specific nature of HIV-1, such as GM95 cells, a mouse-based melanoma cell , use retrovirus to integrate CD4, CXCR4 and CCR5 genes and then transcribe, so that CD4, CXCR4 and CCR5 molecules can be expressed on the cell membrane, which can realize HIV-1 infection, but it is difficult to maintain the complete replication of the virus, so it can only be applied to HIV Intracellular Infection Study of -1

Method used

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  • Mouse cell L1210-based HIV-1 infectible host cell
  • Mouse cell L1210-based HIV-1 infectible host cell
  • Mouse cell L1210-based HIV-1 infectible host cell

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Embodiment Construction

[0028] The principles and features of the present invention are described below in conjunction with examples, which are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0029] 1. Preparation of HIV-1 infectable host cells

[0030] 1) Construction of lentiviral expression vector

[0031] The target genes were hCD4 (GenBank sequence number: NM_000616.4), hCCR5 (GenBank sequence number: NM_000579.3) and human Cyclin T1 (GenBank sequence number: NM_001240.3). A double-promoter lentiviral overexpression vector was constructed from The international standard gene bank was retrieved, and the expression method of bicistronic vector (Bicistronic) was used to prepare the lentiviral packaging vector. The schematic diagram of the structure of the lentiviral vector is shown in figure 1 .

[0032] Using Gateway technology, first construct pDown-CD4-T2A-Cyclin T1, pUP-CMV, pTail-IRES / CCR5; small amount of extraction plasmid pDo...

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Abstract

The invention relates to an application of a mouse leukemia cell L1210 in the preparation of an HIV-1 infectible host cell, and also relates to the HIV-1 infectible host cell. The mouse leukemia cell L1210 is a mouse leukemia cell L1210 with CD4 and CCR5 expressed on the surface of the cell and Cyclin T1 expressed in the cell. The invention also relates to a preparation method of the HIV-1 infectible host cell, and an application of the HIV-1 infectible host cell. The HIV-1 infectible host cell obtained in the invention can be highly-efficiently infected by HIV-1, virus particles replicated in the infected cell has infection activity, and the infected cell secretes active HIV-1 virus to the outside of the cell. The HIV-1 infectible host cell can be used for treating and researching HIV-1 infection and developing anti-HIV-1 drugs.

Description

technical field [0001] The invention relates to the field of HIV treatment and research, and more particularly relates to an HIV-1 infectable host cell and its preparation method and application. Background technique [0002] Human immunodeficiency virus (HIV) is a lentivirus that infects cells of the human immune system and causes acquired immunodeficiency syndrome (AIDS). HIV is divided into type 1 and type 2, and type 1 is the main strain prevalent in the world at present, represented by HIV-1. HIV-1 is highly selective to host cells and can only infect humans and a few primates (such as orangutans, gibbons, etc.). Common monkeys such as rhesus monkeys, macaques and other animals cannot be infected with HIV-1 under normal conditions. [0003] Currently, HIV-1 infection research models mainly include cell models and animal models. The cell model is mainly some CD4-positive cells, such as T4 lymphocytes, mononuclear macrophages, dendritic cells, etc. These cells have CD...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12N5/10C12Q1/68G01N33/68G01N33/571
CPCC07K14/4738C07K14/70514C07K14/7158C12N15/86C12N2740/15043C12Q1/686G01N33/571G01N33/6872C12Q2521/107
Inventor 曾常春李雅婧李丽君何金洋
Owner 深圳市龙华新区中心医院
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