The invention relates to a double-
antigen type 1 diabetes vaccine targeting intestinal M cells and a preparation method of the double-
antigen type 1 diabetes vaccine, and belongs to the technical field of
biology. The method comprises the following steps: connecting human GAD65191-320, INS17-110 and CKS9 targeting
peptide CKSTHPLSC through G4S connecting
peptide according to the sequence, optimizing
lactobacillus codon preference, inserting the human GAD65191-320, the INS17-110 and the CKS9 targeting
peptide CKSTHPLSC into a pNZ8148 vector together, transforming constructed recombinant plasmids into MC1061 competent
escherichia coli for
cloning amplification, and extracting the recombinant plasmids; the recombinant
plasmid is electrically transferred into NZ9000 competent
lactococcus lactis to construct a GAD65191-320 + INS17-110 + CKSTHPLSC recombinant
lactic acid bacteria vaccine, and after amplification,
Nisin is used for inducible expression of the
fusion protein. According to the present invention, the recombinant
lactic acid bacteria vaccine is orally taken to immunize STZ induced BALB / c
diabetes model mice, such that the mice can be induced to produce the GAD65 + INS
specific antibody, and the proliferation of regulatory T cells and dendritic cells secreting
interleukin 10 can be promoted so as to induce the
immune tolerance, inhibit the differentiation of young DCs and maintain the blood glucose level, such that the type 1
diabetes mellitus can be prevented and treated.