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Toxoplasma metabolic secretion antigen vaccine and its prepn process

An antigen vaccine, Toxoplasma gondii technology, applied in the direction of bacterial antigen components, anti-infective drugs, drug combinations, etc., can solve the problems of live vaccines, such as the risk of dispersing toxins, difficulty in extraction, and the lack of research on Toxoplasma gondii vaccine, to achieve effective humoral immunity and The effect of cellular immunity

Active Publication Date: 2007-03-21
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Killed vaccines have no practical value for people and livestock. Live vaccines have the risk of spreading the virus and cannot be popularized and applied. The strains required for attenuated live vaccines need tissue culture cell lines for passage. After long-term preservation, their genetic stability cannot be guaranteed.
(2) Insect body-specific component vaccines, specific components are extracted from insect body lysates or excreted and secreted antigens (ESA) by immunochemical methods, and insect body-specific component vaccines have certain immunity, but there are Problems such as difficulty in extraction
Animal experiments have shown that monovalent subunit vaccines are not ideal, and compound multivalent vaccines are currently being vigorously developed
(4) Nucleic acid vaccines. In terms of nucleic acid vaccines, research on Toxoplasma gondii vaccines has not yet been carried out
So far, there is no Toxoplasma gondii antigen-toxoplasma metabolic secretion antigen compound vaccine

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] On November 5, 2003, 150 mice with a body weight of 20-25 grams were inoculated intraperitoneally with 1×10^5 Toxoplasma tachyzoites per mouse. On November 8, the mice developed symptoms such as depression and rough coat. , the mice were decapitated to death, and immersed in 70% alcohol for body surface disinfection. Each mouse was washed with 2 ml of PH7.2 PBS buffer to collect parasites. A total of 400 ml of peritoneal fluid was collected. The collected peritoneal fluid was centrifuged at 3000 rpm for 30 minutes, and the supernatant was collected. Centrifuge 5 times at 3000 rpm for 30 minutes each time, collect the supernatant, and store it at -20°C for later use.

[0029] At the same time, on November 5, 2003, with the amount of 2 × 10^7 Toxoplasma tachyzoites per bottle, it was inoculated into the vero cells that had grown well and poured the original medium and added a new 100ml serum-free medium. , a total of 35 bottles were inoculated, and the culture was contin...

Embodiment 2

[0033] On February 5, 2004, 100 mice with a body weight of 20-25 grams were inoculated intraperitoneally with 1×10^5 Toxoplasma tachyzoites per mouse. On February 9, the mice developed symptoms such as depression and rough coat. , the mice were killed by decapitation, and then immersed in 75% alcohol for body surface disinfection. Each mouse was washed with 2-3 ml of PH7.2 PBS buffer to collect parasites. A total of 250 ml of peritoneal fluid was collected. The collected peritoneal fluid was centrifuged at 4,000 rpm for 20 minutes, and the supernatant was collected. Centrifuge three times at 4000 rpm for 20 minutes each time, collect the supernatant, and store it at -15°C for later use.

[0034] At the same time, on February 5, 2004, the amount of 2 × 10^7 Toxoplasma tachyzoites per bottle was inoculated into the vero cells that had grown well and poured the original culture solution into a new 100ml serum-free culture solution. , a total of 28 bottles were inoculated, and th...

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PUM

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Abstract

The present invention is toxoplasma metabolic secretion antigen vaccine and its preparation process. The toxoplasma metabolic secretion antigen vaccine is prepared with toxoplasma metabolic secretion antigen. The preparation process includes the following steps: inoculating toxoplasma tachyzoite to abdominal cavity of mouse, killing the mouse and soaking in alcohol for surface sterilizing; collecting polypide; obtaining abdominal cavity liquid via centrifuging and collecting supernatant; inoculating toxoplasma tachyzoite to vero cell to culture and collecting the cultured liquid after cell falling off via centrifuging and collecting supernatant; thrice freezing and thawing abdominal cavity liquid and cultured liquid; mixing, measuring protein content of the mixed liquid; mixing the mixed liquid with M206 adjuvant, emulsifying and packing as the composite toxoplasma vaccine with antigen protein content of 10 mg / ml. The vaccine is for inducing humoral immunity and cellular immunity.

Description

technical field [0001] The invention relates to the field of Toxoplasma gondii vaccines, in particular to a Toxoplasma gondii metabolized and secreted antigen vaccine, and the invention also relates to a preparation method of the vaccine. Background technique [0002] Toxoplasma gondii is an obligate intracellular parasite that can infect all mammals including humans, causing serious damage to human health and the development of animal husbandry. So far, the development of toxoplasma vaccine mainly includes (1) whole worm vaccine, including dead vaccine, live vaccine and live attenuated vaccine. Dead vaccines have no practical value for people and livestock. Live vaccines have the risk of detoxification and cannot be widely used. The strains required for live attenuated vaccines need to be passaged in tissue culture cell lines. After long-term conservation, their genetic stability cannot be guaranteed. (2) Parasite-specific component vaccines, using immunochemical methods t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/02A61P33/02
Inventor 张德林李学瑞王艳华杜重波李慧萍
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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