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Hybridization immune deficiency virus strain and application

An immunodeficiency virus and strain technology, which can be applied to viruses/phages, medical preparations containing active ingredients, and antibody medical ingredients, etc., and can solve problems such as the inability to simulate the natural infection state of HIV-1 well.

Inactive Publication Date: 2007-04-18
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the SHIV constructed in the world use the CXCR4 receptor. After infecting monkeys, they can cause monkeys to become ill within a short period of time. This animal model cannot well simulate the natural infection status of HIV-1 in humans.

Method used

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  • Hybridization immune deficiency virus strain and application
  • Hybridization immune deficiency virus strain and application
  • Hybridization immune deficiency virus strain and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Construction of SHIV, SHIV-B' against the main epidemic strain HIV-1B' subtype in central China WHU Should carry the outer membrane gene (env) of representative B' subtype HIV-1.

[0085] In order to understand that the HIV-1 selected for SHIV construction is the B' subtype strain, an early HIV-1 02HNsmx2 was isolated from an HIV-infected patient in Henan, and the tat of HIV-1 02HNsmx2 was amplified from the cell genome by PCR / rev / vpu / env gene sequence, its sequencing. Molecular analysis of the tat / rev / vpu / env genotype of HIV-1 02HNsmx2 and how it differs from other HIV-1 strains.

[0086] Isolation and culture of peripheral blood mononuclear cells (PBMC) from HIV-1 infected patients and healthy individuals:

[0087] 1. Obtain whole blood (heparin anticoagulated) from HIV-1 infected persons (02HNsmx2) and healthy persons, process or detect tissues, body fluids or excreta that have been separated from the human body or organism to obtain information as an intermediate...

Embodiment 2

[0115] HIV-1 02HNsmx2 tat / rev / vpu / env reading frame integrity and genotype of R5 tropism.

[0116] In order to prove that the open reading frame of each gene of HIV-1 02HNsmx2 tat / rev / vpu / env is complete and has the ability to express proteins in vivo and in vitro, its gene sequence was analyzed to rule out the possibility of not having a complete reading frame. It is necessary to construct an R5-tropic SHIV, so the phenotype of HIV-102HNsmx2 env must be proved to be R5-tropic before constructing SHIV.

[0117] Analysis of HIV-1 02HNsmx2 tat / rev / vpu / env gene reading frame:

[0118] The entire tat / rev / vpu / env gene sequence of HIV-1 02HNsmx2 was compared with the corresponding sequence of the standard strain HIV-1 HXB2 using the ClustalX1.81 program, and the sequence after comparison was submitted to the HIV Databases website ( www.hiv-web.lanl.gov / content / index ). After clicking the Gene Cutter of Tool, the website analyzes the open reading frames of the tat, rev, vpu and en...

Embodiment 3

[0124] Construction of hybrid virus strain SHIV-B' WHU .

[0125] In order to obtain the hybrid virus strain SHIV-B' WHU , the applicant first cloned the 3' end vector carrying the HIV-1B' tat / rev / vpu / env gene, and the vector and the 5' end plasmid were co-transfected into 293T cells to construct a hybrid virus.

[0126] Construction of 3' chimeric vector (3'-p02HN SMX2 -B' WHU ):

[0127] 1. Digest the HIV-1 tat / rev / vpu / env gene fragment from p2'ENV-VAX with restriction enzymes SphI and XhoI, and use the gel recovery kit to recover the exogenous HIV-1 tat / rev / vpu / env gene (3Kb). 3' end vector (3'-pvpu + ) is a non-expression vector (Chen, Z., et al., 2000.Enhanced Infectivity of an R5-Tropic Simian / Human Immunodeficiency Virus Carrying Human Immunodeficiency Virus Type 1 Subtype C Envelope after Serial Passages in Pig-Tailed Macaques (Macaca nemestrina). J Virol, 74: 6501-6510). Digest vector 3'-pvpu with restriction enzymes SphI and XhoI + and dephosphorylated.

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Abstract

This invention relates to a Hybrid immunodeficiency virus strain and application, Human immunodeficiency virus (HIV-1) B' hypotype, R5 tropism, hybrid human / simian immunodeficiency virus strain (SHIV-B'WHU) and 3'end chimeric plasmids used to construct this virus and the use of this virus in stablishing simian / AIDS animal model. At genome framework of simian immunodeficiency virus SIVmac239, gene segment of B'HIV-1 replaces SIVmac239 gene to constitute SHIV-B'WHU. The constituted Hybrid virus SHIV-B'WHU takes along adventitial gene of B'hypotype HIV-1, possessing R5 tropism, it can infect PBMC of human and Macaca rhesus out-of-body and be effectively reproduced, ensuring that SHIV-B'WHU infect Macaca rhesus to construct simian / AIDS animal model. It will be used for researching HIV such as: process of infection in body, etiological agent characteristic, etiopathogenesis,immune reaction and so on, and screening and evaluating AIDS vaccine, anti-HIV drug and microbicide.

Description

technical field [0001] The invention relates to a hybrid immunodeficiency virus derived from human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV), which is characterized by carrying the outer membrane gene of the main epidemic strain HIV-1B' subtype in central China (env); has R5 tropism; has the ability to infect target cells and has the ability to infect human and Chinese rhesus monkey peripheral blood mononuclear cells (PBMC) and replicate in vitro. At the same time, it also involves the application of this hybrid virus. Using it to infect rhesus monkeys can establish a SHIV / monkey model of the B' subtype; to study the in vivo infection process, pathogenic characteristics, pathogenesis, immune response, etc. of HIV, and to detect and Screening for HIV vaccines, antiretroviral drugs and microbicides. Background technique [0002] Human Immunodeficiency Virus (Human Immunodeficiency Virus, HIV) belongs to Retroviridae, Lentiv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N1/21G01N33/53A61K39/21
Inventor 张林琦陈志伟庄柯田波
Owner WUHAN UNIV
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