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Self-transporter used for homologous recombination, and mucosal immune vaccine constructed thereby

A technology of homologous recombination and autotransporter, applied in the field of cervical cancer preventive vaccines, can solve problems such as the stability of foreign genes and the safety of carriers.

Active Publication Date: 2014-11-05
XI AN JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, using attenuated Salmonella as a carrier bacteria to express exogenous protective antigens, but it is not enough to solve the stability of exogenous genes and the safety of vectors

Method used

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  • Self-transporter used for homologous recombination, and mucosal immune vaccine constructed thereby
  • Self-transporter used for homologous recombination, and mucosal immune vaccine constructed thereby
  • Self-transporter used for homologous recombination, and mucosal immune vaccine constructed thereby

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Experimental program
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Embodiment Construction

[0037] The present invention will be further described in detail below in conjunction with specific embodiments, which are explanations of the present invention rather than limitations.

[0038] 1. Selection of autotransporter IcsA

[0039] IcsA is an outer membrane protein with a molecular weight of 120kD, encoded in a virulent plasmid of 220kb, and is the most important factor promoting intercellular transmission of Shigella. As a member of the outer membrane protein autotransport family, IcsA can mediate its own transmembrane and surface presented. Similar to other autotransporters, IcsA has three structural and functional domains: the amino-terminal 52 amino acids (IcsA 1-52) is a signal peptide that mediates the transport of the entire protein across the bacterial inner membrane through the Sec system; the next α domain (Ics 53-757 ) is the main functional domain of the protein. When presented on the surface of the non-dividing segment of bacteria, it can recruit actin ...

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Abstract

The invention discloses a self-transporter used for homologous recombination, and a mucosal immune vaccine constructed thereby. The transporter treats an N1 sequence expressing IcsA amino acid transmembrane signal peptides and a C2 sequence expressing IcsA carboxyl terminal beta areas as homologous arms, and can be applied to donor vectors for the homologous recombination of Shigella icsA. The stable existence of antigen genes in a vector genome and the surface presence of antigens are realized through integration of antigen proteins of exogenous HPV (human papilloma virus) into self-transport proteins of intestinal bacteria, so a vaccine capable of inducing an effective mucosal immune reaction is exploited, and a system immune reaction and a mucosal immune reaction are simultaneously stimulated. Recombinant attenuated Shigella subjected to the mucosal immunization can effectively excite the generation of HPV16 L1 specific IgG in a serum, and the presence of HPV16 L1 specific antibody secreting cells in the spleen in a high frequency manner.

Description

technical field [0001] The invention belongs to the technical field of cervical cancer prevention vaccines, and relates to an autotransporter for homologous recombination and a vaccine for transmucosal immunity constructed thereof. Background technique [0002] Since most microbial infections occur in the mucosa, the body induces protective humoral immunity and cellular immune responses in the mucosal immune tissue, and it is difficult to induce effective mucosal immune responses through vaccines through non-mucosal immune pathways. Research on the ideal mucosal system protective antigen transport carrier become the focus of vaccine research. [0003] It has been reported that the use of bacteria and viruses that grow and multiply in the attenuated gut or gut-related tissues can more effectively stimulate local and systemic immune responses. For example, the use of attenuated Salmonella as a carrier bacteria expresses exogenous protective antigens, but it is not enough to s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/09A61K48/00A61P35/00A61P31/20
Inventor 王一理于军许丹杨筱凤
Owner XI AN JIAOTONG UNIV