Enhanced bivalent DC (dendritic cell) vaccine of human papillomavirus HPV-16/18

A technology of human papillomavirus and HPV-16, applied in the field of biology and new medicine, can solve the problems of large amount of infected tissue, difficulty in determining the optimal stimulation dose, and induction of autoimmune reactions

Inactive Publication Date: 2017-07-21
SHANDONG XINRUI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this scheme requires a large amount of infected tissue, and the optimal stimulation amount is difficult to determine
In addition, virus-infected cells also contain the body's own antigens, which have the risk of inducing an autoimmune response

Method used

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  • Enhanced bivalent DC (dendritic cell) vaccine of human papillomavirus HPV-16/18
  • Enhanced bivalent DC (dendritic cell) vaccine of human papillomavirus HPV-16/18
  • Enhanced bivalent DC (dendritic cell) vaccine of human papillomavirus HPV-16/18

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Induction of imDC

[0032] (1) Collect 50 ml of umbilical cord blood from healthy puerperas, use lymphocyte separation medium to separate mononuclear cells, and culture medium (purchased from TaKaRa Company, GT-T551), 37 ° C, 5% CO 2After culturing under the condition for 2-3 hours, discard the suspended dead cells, keep the adherent cells, and add cytokines rhIL-4 (final concentration: 50ng / ml), rhGM-GSF (final concentration: 100ng / ml) to induce single Differentiate nuclear cells into DCs, replace the new medium every 48 hours, obtain imDCs on the fifth day of culture, observe the cell morphology of DCs before maturation through an inverted microscope, detect the expression of cell surface molecular markers CD83+ and CD86+ by flow cytometry, and comprehensively identify the induced imDC.

Embodiment 2

[0033] Example 2: L1 sequence modification of HPV-16 and HPV-18 mature DC cells

[0034] (1) Packaging and preparation of lentivirus: using gene cloning technology, PCR amplifies the full-length cDNA sequence of L1 gene of HPV-16 and HPV-18, and connects it into the lentiviral expression vector pLent-C-GFP to construct the lentivirus of L1 antigen. Viral expression vector, identified by PCR and nucleic acid sequencing. Perform virus titer determination.

[0035] (2) Infect imDC with the constructed L1 lentiviral vector, culture the infected imDC for 6 hours, add the maturation factor TNF-α, induce mature DC cells, observe the cell morphology of mature DC through an inverted microscope, and detect the cells by flow cytometry Expression of surface molecular markers CD83+ and CD86+ comprehensively identifies induced mDCs. The mDC can be directly injected as a vaccine against HPV16 and HPV18.

[0036] In order to verify the presence or absence of GM-CSF on the expression of HPV...

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Abstract

The invention discloses an enhanced bivalent DC (dendritic cell) vaccine of human papillomavirus HPV-16/18, comprising genetically-modified dendritic cells, recombinant gene sequences in the genetically-modified dendritic cells are serially connected via granulocyte-macrophage stimulating factor and L1 transcription regions of human papillomavirus HPV-16/18, and the DC vaccine is composed of genetically-modified DC surface antigen, comprising granulocyte-macrophage stimulating factor expressed via an enhancement antigen, and L1 antigen proteins of human papillomavirus HPV-16/18. The DC vaccine is a prevention and treatment vaccine, capable of effectively preventing HPV-16 and HPV-18 infections and related diseases caused by HPV infections.

Description

technical field [0001] The invention relates to the technical field of biology and new medicine, more specifically, it is a bivalent DC vaccine of enhanced human papillomavirus HPV-16 / 18. Background technique [0002] Human papillomavirus (Human papillomavirus, HPV) is a papillomavirus A genus belonging to the Papovaviridae family. It is easy to infect human epidermis and mucosal squamous epithelium. It is the cause of cervical cancer, genital warts and some anal cancers. leading cause of oral cancer. Persistent infection of high-risk HPV-16 and HPV-18 may cause cervical cancer, rectal cancer, oral cancer, tonsil cancer and other malignant tumors. Cervical cancer is currently the only malignant tumor with a clear etiology. 99.7% of cervical cancer cases are caused by HPV infection. The World Health Organization and the International Cancer Center have also confirmed that persistent infection of high-risk HPV viruses is the main cause of cervical cancer. People habitually b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/295A61K38/19A61P31/20A61P35/00
CPCA61K38/193A61K39/12A61K2039/5156A61K2039/70C12N2710/20034A61K2300/00
Inventor 刘明录李言志万磊强邦明冯建海金海峰
Owner SHANDONG XINRUI BIOTECH CO LTD
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