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T cell vaccine constructed by secretory component of gene engineering-based aAPC (artificial Antigen Presenting Cell) as well as preparation method and application thereof

A technology of genetic engineering and cell secretion, which is applied in the field of preparation of T cell vaccines and can solve the problems of lack of endogenous expression

Inactive Publication Date: 2018-10-02
项雯华
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because its expression of CD54 and LFA-3 helps to form an effective immune synapse to promote T cell activation [11,12], while its lack of endogenous expression of HLA-A, B and DR molecules limits its allogeneic Side effects caused by T cell stimulation

Method used

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  • T cell vaccine constructed by secretory component of gene engineering-based aAPC (artificial Antigen Presenting Cell) as well as preparation method and application thereof
  • T cell vaccine constructed by secretory component of gene engineering-based aAPC (artificial Antigen Presenting Cell) as well as preparation method and application thereof
  • T cell vaccine constructed by secretory component of gene engineering-based aAPC (artificial Antigen Presenting Cell) as well as preparation method and application thereof

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Embodiment Construction

[0039] The present invention shows that activated non-specific T cells can acquire vesicles derived from genetically engineered artificial antigen-presenting cells, in particular, these cells can acquire antigen-specific histocompatibility complexes and co-stimulatory molecules from the vesicles. The present invention shows that these vesicle-derived molecules are functional. In this way, non-specific T cells targeted by vesicles can directly stimulate antigen-specific immune responses.

[0040] The present invention shows that the genetically engineered artificial antigen-presenting cells are derived from K562 cells that lack endogenously expressed HLA-A, B and DR histocompatibility conformers. K562 cells were first expressed by two eukaryotic expression plasmid vectors pcDNA Hygro / HLA-A2 and

[0041] pcDNA Neo / CD80 transfection. AdV Gag and AdV 41BBL Transfection to form genetically engineered antigen-presenting cells K562 expressing transgenic HLA-A2, CD80, Gag and 41...

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Abstract

The invention discloses a secretory component vesica of a gene engineering-based aAPC (artificial Antigen Presenting Cell), as well as preparation and application of a T cell vaccine constructed by anexosome. By transfecting two kinds of eukaryotic expression plasmids of pcDNAHLA-A2 and pcDNACD80 and infecting two kinds of recombinant adenovirus carriers of AdVGag and AdV41BBL, a gene engineering-based K562A2 / Gag / CD80 / 41BBLaAPC is constructed; then exosomes are purified in K562A2 / Gag / CD80 / 41BBL culture supernatant through a differential superspeed centrifugal method, and electron microscopingand immunoblotting analysis can be carried out, wherein the exosomes are in co-incubation with nonspecific T cells of Con-A stimulated HLA-A2 transgenic mice through incubating, so that Gag-Texo vaccine having HLA-A-A2 limitation and Gag specificity is constructed. According to the secretory component vesica disclosed by the invention, the aAPCK562A2 / Gag / CD80 / 41BBL vesica in unlimited source andcontinuous growth is used for replacing a DC vesica in limited source and is used for preparing an HIV-1 (Human Immunodeficiency Viru I) Gag specific T cell vaccine. A novel method for preparing the Tcell vaccine by using the gene engineering-based aAPC has important influence on development of clinic therapeutic vaccines for a patient suffering HIV-1.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a preparation method and application of a T cell vaccine constructed from secretory bodies of genetically engineered artificial antigen-presenting cells. Background technique [0002] Acquired immunodeficiency syndrome (AIDS) in patients infected with human immunodeficiency virus-I (HIV-I) is a rapidly expanding global pandemic disease. Effective antiviral therapy (HAART) can successfully inhibit the replication of the virus and significantly improve the prognosis of patients [1]. However, it did not restore the anti-HIV-1 immune response. Drug treatment during the chronic phase of inflammation does not reverse virus-induced CD4 + T cells and CD8 + Decline of effector T cells, leading to latent HIV-1 virus [2]. Therefore, it is of practical significance to urgently develop new therapeutic strategies to improve virus control during drug intermittent treatment, so that pa...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/12A61P31/20A61P35/00A61P37/06C12N5/0783C12N5/0784
CPCA61K39/0008A61K39/0011A61K39/12A61K2039/5158A61K2039/572C12N5/0636C12N5/0639C12N2740/16034
Inventor 项建华
Owner 项雯华
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