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Method for inducing immune response using HBs DNA vaccine inoculated by HBV core protein reinforcing gene gun

A core protein, hepatitis B virus technology, applied in gene therapy, antiviral agents, pharmaceutical formulations, etc.

Inactive Publication Date: 2005-06-22
FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no research report on the application of HBc-enhanced gene gun inoculation of HBs DNA vaccine to induce Th1 immune response, and there is no invention patent similar to this strategy

Method used

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  • Method for inducing immune response using HBs DNA vaccine inoculated by HBV core protein reinforcing gene gun
  • Method for inducing immune response using HBs DNA vaccine inoculated by HBV core protein reinforcing gene gun
  • Method for inducing immune response using HBs DNA vaccine inoculated by HBV core protein reinforcing gene gun

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1D

[0046] Expression of Example 1 DNA Vaccine Vector in Mammalian Cells

[0047] Plasmids pIRES / S, pIRES / S / Core and pIRES / S / C149 were transiently transfected into HepG2 cells by calcium phosphate method, and the expression of HBsAg in the supernatant of transfected cells was detected by ELISA 48 hours later. The control was the supernatant of untransfected cells. The results are shown in Figure 2A, pIRES / S, pIRES / S / Core and pIRES / S / C149 can all express HBsAg, the OD450nm of each group is 0.832±0.349, 0.57±0.162 and 0.561±0.277, and the control is 0.052±0.024.

[0048] The plasmids IRES / Core, IRES / S / Core, IRES / C149, and IRES / S / C149 were transiently transfected into Cos7 cells by liposome method, and 48 hours later, anti-HBV Core monoclonal antibody and Rhodanmin-labeled goat anti-mouse secondary antibody were used For immunofluorescence staining. Laser confocal scanning results showed that Core protein and its truncated C149 protein were stained with red fluorescence by Rhodanmi...

Embodiment 2

[0049] HBcAg-specific humoral immunity level after embodiment 2 immunization

[0050] BALB / c mice were randomly divided into 8 groups, 6 mice in each group. The first booster was given in the 4th week after the initial immunization, and the second booster was given in the 8th week. Two weeks after the second booster (10 Week) HBcAg-specific IgG level and IgG subclass level in serum. The control was untreated mouse serum. The results are shown in Figure 3. All groups inoculated with plasmids expressing full-length HBcAg or HBcAg1-149 truncated forms could induce HBcAg-specific IgG antibodies, and there was no statistical difference among the groups. Further analysis of the subclasses in each group found that the anti-HBcAg subclasses of pIRES / Core, pIRES / S / Core and pIRES / Core+pIRES / S expressing full-length HBcAg were mainly IgG2a, and their IgG2a / IgG1 ratios were 2.47 and 2.73 respectively. and 2.07, while the anti-HBcAg subclasses of pIRES / C149+pIRES / S and pIRES / S / C149 ex...

Embodiment 3

[0051] HBsAg-specific humoral immunity level after embodiment 3 immunization

[0052] In order to monitor the change of anti-HBsAg total IgG titer, the gene gun was inoculated with pIRES / S, pIRES / S / Core, pIRES / Core+pIRES / S, pIRES / S / C149, pIRES / C149+pIRES / S, intramuscular injection Serum collected from pIRES / S and untreated control groups were mixed in equal volumes in each group, and the titer level was detected by ELISA method. The results are shown in Figure 5A. At the 14th week, the antibody titers in each group were 1:4800, 1:4800, 1:4800, 1:4800 and 1:6400, respectively. The serum levels of HBsAg-specific IgG were detected two weeks after the second booster (week 10) in each group, and the results are shown in FIG. 5B . In addition to the pIRES / Core group and the control group, pIRES / S, pIRES / S / Core, pIRES / Core+pIRES / S, pIRES / C149+pIRES / S, pIRES / S / C149 and intramuscular injection of pIRES / S group can induce HBsAg-specific IgG antibody, there is no statistical differen...

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PUM

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Abstract

Method for inducing TH1 type immune response by inoculating HBs DNA vaccine through using HBV core protein reinforced gene gun, wherein co-leading / co-expression HBV Core gene is used as adjuvant simultaneously while inoculating plasmid HBs DNA vaccine with a gene gun. The animal immunity experiment shows that, the invention can be applied to reinforce the TH1 type immune response level induced through gene gun vaccinated HBs DNA vaccine, including total IgG, IgG2a, specific CTL activity, and antigen specific IFN-gamma production capacity.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a method for enhancing the Th1 type immune response induced by gene gun inoculation of HBs DNA vaccine with core protein of hepatitis B virus and its application. Background technique [0002] DNA vaccine (DNA vaccine) is considered to be a new direction of future vaccine development because it can overcome the defects of traditional vaccines, induce the body to produce cellular and humoral immune responses at the same time, and has the advantages of simple preparation and stability. Gene gun inoculation of DNA vaccines has great application prospects in human immunity because of the characteristics that only a small amount of plasmid DNA can induce a strong immune response (Kaiserlian D et al. Eur J Dermatol 1999, 9: 169-76; Stringl G et al. Immunol ser 1989, 46: 3-72; Chen D et al. Expert Rev Vaccines 2002, 1: 265-76; Fynan EF et al. Proc Natl Acad Sci USA 1993, 90: 11478-82)...

Claims

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Application Information

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IPC IPC(8): A61K39/39A61K48/00A61P1/16A61P31/12
Inventor 袁正宏周晓辉
Owner FUDAN UNIV
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