DNA vaccine for preventing toxoplasmosis of humans or animals

A DNA vaccine and toxoplasmosis technology, applied in the field of vaccines, can solve the problems of different immune protection effects, the ROP2DNA vaccine immune protection effect cannot reach a satisfactory level, and the ROP is less, so as to improve the immune protection response and survival rate. , Improve the immune response and survival rate, reduce the effect of the formation rate

Inactive Publication Date: 2016-05-11
SHANDONG UNIV
View PDF5 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been some reports on the vaccine research of ROP family members, mainly including ROP1 and ROP2 families. The ROP2 family members that have been confirmed as vaccine candidate genes include ROP2, ROP4, ROP5, ROP8, ROP13, ROP16, ROP17 and ROP18. The amino acid sequences and specific structural units of the rod-shaped proteins are different, resulting in different immune principles and different immune protection effects.
[0004] The article "Immuneresponse induced by recombinant Mycobacterium bovis BCG expressing ROP2 gene of Toxoplasmagondii" published in the SCI journal Parasitology International publicly reported the research report

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • DNA vaccine for preventing toxoplasmosis of humans or animals
  • DNA vaccine for preventing toxoplasmosis of humans or animals
  • DNA vaccine for preventing toxoplasmosis of humans or animals

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0032] Example 1. Toxoplasma gondii ROP19 epitope prediction

[0033] Predict the B cell dominant epitope of ROP19 (ToxoDB10.0(http: / / toxodb.org / toxo / )(GeneID:TGME49_242240)) by comprehensive analysis of DNAStar-Protean software; use IEDB to predict both human and BALB / C mice Toxoplasma gondii ROP19 T cell dominant epitope restricted by MHC molecules ( figure 2 ).

example 2

[0034] Example 2. DNA vaccine of recombinant Toxoplasma gondii ROP19 gene

[0035] According to the gene sequence of Toxoplasma gondii ROP19, the synthetic primers were designed as follows: upstream primers:

[0036] 5'-CGGGGTACCATGAGAAGGCTGCTGCTTTC-3', as shown in SEQ ID NO: 1; downstream primer:

[0037] 5'-CGGGATCCTCACTGAGATCTGGATGC-3', as shown in SEQ ID NO:2.

[0038] Such as figure 1 As shown, the extracted Toxoplasma gondii genomic DNA was used as a template to amplify the ROP19 gene, and the reaction conditions were: denaturation at 95°C for 30s, annealing at 55°C for 30s, extension at 72°C for 3min, a total of 30 cycles. The amplified product was subjected to 1% agarose gel electrophoresis, and the gel was cut to recover the target fragment. The recovered PCR product and the pEGFP-C1 plasmid vector were respectively digested for 3 hours and then electrophoresed. After cutting the gel, the kit was used to recover the digested PCR product The purified product and th...

example 3

[0039] Example 3. Toxoplasma gondii recombinant gene vaccine immunized BALB / c mice

[0040] SPF grade female BALB / c mice (6-8 weeks) were purchased from the Experimental Animal Center of Shandong University. Forty-five mice were randomly divided into 3 groups. The mice in the experimental group were injected with 100 μg of the recombinant vaccine pEGFP-C1-ROP19 through the hind leg muscles, and the mice in the control group were injected with 100 μg of the empty vector pEGFP-C1 and 100 μg of PBS. Mice were immunized three times with two weeks apart.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a DNA vaccine for preventing toxoplasmosis of humans or animals. An ROP19 gene is amplified through PCR, the amplified ROP19 gene is inserted into a eukaryotic expression vector pEGFP-C1 to build recombinant plasmid pEGFP-C1-ROP19 (pROP19), the recombinant plasmid containing a target gene is extracted, and the toxoplasma gondii ROP19DNA vaccine is obtained. According to the DNA vaccine for preventing the toxoplasmosis of the humans or the animals, active immunization is conducted on BALB/c mice through the DNA vaccine, the immunogenicity of the vaccine is evaluated by determining cells and humoral immunity indexes of immunized mice, and the immunoprotectivity of the vaccine is estimated by counting the number of cysts in brains of the mice and the survival rate of the mice after attack experiments of toxoplasma gondii are conducted. By means of the DNA vaccine for preventing the toxoplasmosis of the humans or the animals, humoral immunity and cellular immune responses can be effectively enhanced, the formation rate of the brain cysts of the immune mice attacked by a toxoplasma gondii PRU strain (II type) is effectively decreased, and the survival time of the mice attacked by a toxoplasma gondii RH stain (I type) is prolonged.

Description

technical field [0001] The present invention relates to a vaccine, in particular to a compound antigenic epitope based on toxoplasma gondii rod protein 19 (ROP19), a toxoplasma vaccine obtained by constructing a recombinant plasmid containing a target gene, used for human or animal toxoplasmosis prevention. Background technique [0002] Toxoplasma gondii (Toxoplasmagondii) is a worldwide distribution of parasitic protozoa, widely parasitic in nucleated cells of humans and animals, causing severe zoonotic diseases. Toxoplasma infection in early pregnancy women can lead to miscarriage, premature birth or even stillbirth, and can also cause neonatal malformations and fetal eye complications through the placenta. Toxoplasma gondii can lead to severe clinical symptoms (such as toxoplasmic encephalopathy) and even death for people with weak or weak immunity (such as AIDS and tumor patients). In immunocompetent hosts, Toxoplasma tends to form chronic infections, causing irreversi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A61K48/00A61K39/012A61P33/02C12N15/12C12N15/79
CPCA61K39/012A61K2039/53C07K14/45C12N15/79C12N2800/106
Inventor 周剑何深一吕刚王琳赵晗婷
Owner SHANDONG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap