DcR3 (Decoy Receptor 3) and GAD65 (Glutamic Acid Decarboxylase 65) double gene co-expression recombinant adenovirus as well as preparation method and application thereof

Abstract

The invention discloses a DcR3 (Decoy Receptor 3) and GAD65 (Glutamic Acid Decarboxylase 65) double gene co-expression recombinant adenovirus as well as a preparation method and application thereof. The recombinant adenovirus genome comprises a DcR3 and GAD65 double gene expression cassette which sequentially comprises a CMV (Cytomegalovirus) promoter, a DcR3 complete encoding gene, a terminator, an internal ribosomal entry site (IRES), a GAD65 complete encoding gene and a terminator from upstream to downstream; the recombinant adenovirus has simple preparation method; and after the recombinant adenovirus is transfected with a dendritic cell in vitro and then subjected to reinfusion to be in vitro, the recombinant adenovirus can induce the tolerance of specific T cells, effectively inhibit the damage of islet beta cells and prolong the life cycle of a transplanted islet; and the recombinant adenovirus can be used for preparing a dendritic cell vaccine for resisting islet transplantation rejection and has favorable development and application potentials in the field of diabetes mellitus treatment.

Description

technical field [0001] The invention relates to a recombinant adenovirus, in particular to a DcR3 and GAD65 double-gene co-expression recombinant adenovirus, and also relates to a preparation method and application of the recombinant adenovirus. Background technique [0002] With the improvement of people's living standards and changes in lifestyle, the incidence of diabetes is increasing day by day, and it has become the third major disease that affects people's health and life after cardiovascular disease and tumors. Islet transplantation is simple, safe, and effective. Postoperatively, patients do not need to rely on exogenous insulin to correct the disorder of glucose metabolism in the body and significantly improve the quality of life. It has attracted much attention in the treatment of diabetes, especially type 1 diabetes. However, due to the difference in the genetic background of the donor and the recipient, islet transplantation is susceptible to immune rejection b...

AI Technical Summary

Problems solved by technology

[0004] There are two main methods of gene combination therapy: one is to simultaneously transfect target cells with a variety of recombinant expression vectors carrying different genes, which can be freely adjusted The proportion of each recombinant expression vector, but the recombinant expression vectors carrying different genes need to be constructed separately, the construction work is cumbersome, time-consuming, and there are many influencing factors; in addition, due to the randomness of the transfection process, the cells after transfection have uneven gene copies It is not conducive to the expression of the target gene and subsequent treatment, but also makes it difficult to evaluate and analyze the experimental results; the above shortcomings limit the further application of this method

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