Coronavirus-targeting universal DC cell vaccine, and preparation method and use thereof

A method for preparing a coronavirus-targeting universal DC cell vaccine, comprising the following steps:

• (1) ligating a fusion gene onto an expression vector to obtain a recombinant vector,
• wherein the fusion gene comprises a first fusion gene or a second fusion gene, the first fusion gene comprises an HLA gene and a gene of coronavirus open reading frame protein connected by a linker sequence, and the second fusion gene comprises an HLA gene and a gene of coronavirus structural protein connected by a linker sequence;
• (2) transferring the recombinant vector of step (1) into antigen-presenting cells to be transfected, and using antibiotics to screen antigen-presenting cells into which the recombinant vector has been transferred[[ ]], so as to obtain the
• coronavirus-targeting universal DC cell vaccine;
• wherein in step (1), the gene of coronavirus open reading frame protein is selected from
• one of ORF3a, ORF6, ORF7a and ORF8;
• the nucleotide sequence of ORF3a is SEQ ID NO:
• 1;
• the nucleotide sequence of ORF6 is SEQ ID NO:
• 2;
• the nucleotide sequence of ORF7a is SEQ ID NO:
• 3;
• and the nucleotide sequence of ORF8 is SEQ ID NO:
• 4;
• wherein the gene of the coronavirus structural protein in step (1) is selected from one of a coronavirus E protein gene, a coronavirus S protein gene, a coronavirus PLP protein gene, a coronavirus M protein gene and a coronavirus N protein gene;
• the nucleotide sequence of coronavirus E protein gene is SEQ ID NO:
• 5;
• the nucleotide sequence of coronavirus S protein gene is SEQ ID NO:
• 6;
• the nucleotide sequence of coronavirus PLP protein gene is SEQ ID NO:
• 7;
• the nucleotide sequence of coronavirus M protein gene is SEQ ID NO:
• 8;
• and the nucleotide sequence of coronavirus N protein gene is SEQ ID NO:
• 9;
• and wherein the method for preparing the antigen-presenting cells to be transfected in step (2) comprises the following steps:
• (a) ligating a fusion gene of TAX and ST40 connected by a linker sequence onto an expression vector to obtain a recombinant vector, wherein the nucleotide sequence of TAX is SEQ ID NO:
• 11, and the nucleotide sequence of ST40 is SEQ ID NO:
• 12;
• (b) packaging a lentivirus with the recombinant vector of step (a) to obtain a packaged virus;
• (c) infecting DC cells with the packaged virus of step (b) to obtain infected DC cells;
• (d) inoculating trophoblast cells in an upper culture chamber of a cell incubator, inoculating the infected DC cells obtained in step (c) in a lower culture chamber of the cell incubator, conducting first culture for 4-6 weeks, conducting second culture for 1-2 weeks after removing CD3+ cells, then conducting third culture for more than 2 months after removing CD3+ cells to obtain the antigen-presenting cells to be transfected.