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Peptide-DNA double vaccine based on T-cell epitope for anti-Schistosoma japonicum infection

A technology of epitope peptide and schistosomiasis, applied in the field of immunology, can solve the problem of unclear T epitope and achieve the effect of reducing cross-reaction

Inactive Publication Date: 2009-10-28
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the specific MHC high-affinity T epitopes in the aforementioned existing schistosomiasis vaccine candidate molecules are still unclear
However, there is no report on the use of T cell epitopes to construct a peptide-DNA double vaccine against Schistosoma japonicum infection

Method used

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  • Peptide-DNA double vaccine based on T-cell epitope for anti-Schistosoma japonicum infection
  • Peptide-DNA double vaccine based on T-cell epitope for anti-Schistosoma japonicum infection
  • Peptide-DNA double vaccine based on T-cell epitope for anti-Schistosoma japonicum infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Preparation of PDDV

[0034] Using various software such as SYFPEITHI and TEPITOPE to predict and analyze the T cell epitope of the full-length antigen of Schistosoma japonicum 22.6kDa surface membrane protein, and then select the epitope with the highest score after homology comparison and restriction of restriction enzyme sites. position, its amino acid sequence is AKQYNICCKFKELLD (Ala Lys Gln Tyr Asn Ile Cys Cys Lys Phe Lys Glu Leu Leu Asp).

[0035] Build process like figure 1 as shown,

[0036] 1) Order the peptide of the above sequence from Invitrogen and add 18 lysines KKKKKKKKKKKKKKKKKKAKQYNICCKFKELLD (purity ≥ 99%) to its amino terminus during synthesis, so that the entire synthetic peptide is positively charged and named P5. In addition, to set up a control group, we also ordered a peptide segment (purity ≥ 99%) containing only 18 lysines KKKKKKKKKKKKKKKKKK, and named it 18K.

[0037] 2) Add a Sal I restriction site to the 5' end of the epitope p...

Embodiment 2

[0061] Embodiment two: carry out immunity / challenge infection experiment with the PDDV vaccine that embodiment one makes

[0062] Clean grade C57BL / 6 mice: purchased from the National Mouse Genetic Resources Celebration Center of Nanjing University, 8 weeks old, female.

[0063] First experiment:

[0064] A) The mice were randomly divided into three groups, 14 in each group, named as T5-PDDV group, 18K-PDDV group and control group respectively.

[0065] B) Immunize once a week, subcutaneously inject 100 μl of T5-PDDV solution (containing 28 μg P5 peptide segment and 5 μg recombinant eukaryotic expression plasmid DNA) and 18K-PDDV solution (containing 28 μg 18K peptide segment and 5 μg empty eukaryotic expression plasmid DNA) or PBS solution.

[0066] C) Detection of the immune response induced by PDDV vaccine in mice: ① before immunization and 2 weeks after the last immunization, blood samples were taken from the tail vein of the mice, and used for ELISA to detect the seru...

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Abstract

The invention belongs to the field of immunology, and in particular relates to a mixed vaccine of T cell epitope protein and DNA against Schistosoma japonicum infection. The said anti-Schistosoma japonicum infection peptide-DNA double vaccine is to wrap the protein containing the epitope peptide outside the eukaryotic expression vector with the gene encoding the epitope peptide, the amino acid sequence of the said T cell epitope peptide For AKQYNICCKFKELLD. The peptide-DNA double vaccine can be prepared by cationic peptide loading technology. The anti-Schistosoma japonicum infection peptide-DNA double vaccine of the present invention can effectively resist the degradation of DNase in the body after injection into the body, thereby greatly reducing the dosage of the vaccine; it is easy to enter the antigen-presenting cells and the antigen is efficiently presented; Facilitates simultaneous stimulation of cellular and humoral immune responses.

Description

technical field [0001] The invention belongs to the field of immunology, in particular to a protein and DNA mixed vaccine against Schistosoma japonicum infection. Background technique: [0002] In recent years, the epidemic of schistosomiasis in my country has rebounded again, and the urgent need to rely on scientific and technological progress to consolidate and improve the effectiveness of schistosomiasis control has once again been put on the agenda. In view of the unparalleled role of vaccines in the control of many infectious diseases, trying to develop anti-schistosomiasis vaccines to obtain the dual benefits of combining the "short-acting" effect of chemotherapy and the "long-acting" immune prevention effect induced by vaccination to solve schistosomiasis The expectation of control is the goal that relevant scientists around the world have been striving for in the past 20-30 years, and it is also the original intention of WHO / TDR to coordinate the development of schis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61P33/12
Inventor 苏川张兆松吴观陵
Owner NANJING MEDICAL UNIV
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