Pseudoviruses for non-CD4 receptors of HIV-1 CRF_BC recombinant strain and application of pseudoviruses

A technology of HIV-1, 1.HIV-1, which is applied to the non-CD4 receptor pseudovirus of HIV-1 CRF_BC recombinant strain and its application field, and can solve the problems such as unclear mechanism

Active Publication Date: 2015-09-02
BEIJING YOUAN HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These studies have shown that the popular CRF_BC recombinant strains in my country are not only biologically different from other subtypes, but also in the sensitivity of neutralizing antibodies. The mechanism is still unclear, so it is necessary to target CRF_BC Envelope proteins of recombinant strains were studied in depth

Method used

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  • Pseudoviruses for non-CD4 receptors of HIV-1 CRF_BC recombinant strain and application of pseudoviruses
  • Pseudoviruses for non-CD4 receptors of HIV-1 CRF_BC recombinant strain and application of pseudoviruses
  • Pseudoviruses for non-CD4 receptors of HIV-1 CRF_BC recombinant strain and application of pseudoviruses

Examples

Experimental program
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Embodiment 1

[0096] The preparation of the pseudovirus that the non-CD4 receptor of embodiment 1, HIV-1 CRF_BC recombinant strain utilizes

[0097] 1. Site-directed mutation of the envelope protein gene of the non-CD4 receptor-utilized pseudovirus of the HIV-1 CRF_BC recombinant strain

[0098] Use Stratagen's mutation kit (Quick Change site-directed mutagenesis kit) to carry out 197 site N (asparagine Asn) to S (serine Ser) on the envelope protein gene of HIV-1 CRF07_BC enveloped pseudovirus CH120.6 The point mutation of the mutated envelope protein gene shown in SEQ ID No.3 is obtained. The expressed amino acid sequence of the mutated envelope protein gene is envelope protein A of SEQ ID No.1. The mutated envelope protein gene The membrane protein gene was named envelope protein A gene.

[0099]Use Stratagen's mutation kit (Quick Change site-directed mutagenesis kit) to carry out 194 site N (asparagine Asn) to S (serine Ser) on the envelope protein gene of HIV-1 CRF07_BC enveloped pseud...

Embodiment 2

[0110] Example 2, Verification and infectivity experiments of pseudoviruses utilized by non-CD4 receptors of HIV-1 CRF_BC recombinant strains

[0111] The non-CD4 receptor pseudovirus CH120N197S of the HIV-1 CRF_BC recombinant strain prepared in Example 1 was used to infect target cells Cf2Th-CD4 / CCR5 with CD4 receptors and target cells Cf2Th-CCR5 without CD4 receptors respectively, HIV-1 CRF07_BC enveloped pseudovirus CH120.6 was respectively infected with target cell Cf2Th-CD4 / CCR5 with CD4 receptor and target cell Cf2Th-CCR5 without CD4 receptor as a control; HIV-1 prepared in Example 1 The non-CD4 receptor pseudovirus CH110N197S of the CRF_BC recombinant strain infected target cells Cf2Th-CD4 / CCR5 with CD4 receptors and target cells Cf2Th-CCR5 without CD4 receptors respectively, and the HIV-1CRF07_BC enveloped pseudovirus CH110 .2 Respectively infect target cells Cf2Th-CD4 / CCR5 with CD4 receptors and target cells Cf2Th-CCR5 without CD4 receptors as controls; at the same ti...

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Abstract

The invention discloses pseudoviruses for non-CD4 receptors of an HIV-1 CRF_BC recombinant strain and application of the pseudoviruses. The invention successfully constructs the pseudoviruses CH120N197S and CH110N197S for the non-CD4 receptors of the HIV-1 CRF_BC recombinant strain, and the two pseudoviruses not only can infect a target cell with the CD4 receptor, but also can infect a target cell without the CD4 receptor. The infectivity of the pseudovirus CH120N197S infecting the target cell without the CD4 receptor is 73/100 of that of the pseudovirus CH120N197S infecting the target cell with the CD4 receptor and is higher than that of a reported pseudovirus e7adaN197S (60/100). The successfully-constructed pseudoviruses for the non-CD4 receptors of the HIV-1 CRF_BC recombinant strain greatly enrich the main HIV-1 pseudovirus resources in China so as to have very important significance for studying the structural characteristics of cell membrane proteins of the CRF_BC recombinant strain serving as a main epidemic strain in China and developing an HIV vaccine for the CRF_BC recombinant strain in China.

Description

technical field [0001] The invention relates to a pseudovirus utilized by non-CD4 receptors of HIV-1 CRF_BC recombinant strains in the field of biomedicine and an application thereof. Background technique [0002] HIV-1 viral envelope protein is a glycoprotein composed of 850-880 amino acids encoded by the env gene (gpl60). There is an arginine-rich hydrophobic region in gp160, which can be cleaved by host proteases into two subunits, gp120 and gp41. The CD4 receptor binding region on the HIV-1 viral envelope protein gp120 first binds to the CD4 receptor on the target cell, completing the first step of HIV-1 virus invasion. Then the co-receptor binding region on the envelope protein gp120 binds to the co-receptors (CCR5 and / or CXCR4) on the target cells, completing the second step of virus invasion. Finally, the HIV-1 viral envelope protein gp41 mediates the fusion of the viral envelope with the target cell plasma membrane, completing the process of virus infection and inv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01C12N5/10C07K14/005C12N15/33C12N15/86A61K35/76A61K39/12A61P31/18
Inventor 孙坚萍张永宏孙焕芹李宁
Owner BEIJING YOUAN HOSPITAL CAPITAL MEDICAL UNIV
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