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Listeria monocytogenes inactivated vaccine and preparation thereof

A technology of Listeria and monocytes, applied in the direction of antibacterial drugs, bacterial antigen components, etc., can solve the problems of short production cycle and so on

Inactive Publication Date: 2008-09-17
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the production of inactivated vaccines by irradiation is simple, safe, low in cost, and short in production cycle, but there is no relevant report so far

Method used

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  • Listeria monocytogenes inactivated vaccine and preparation thereof
  • Listeria monocytogenes inactivated vaccine and preparation thereof
  • Listeria monocytogenes inactivated vaccine and preparation thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Bacteria inactivated by irradiation

[0022] Bacterial preparation: yzu LM 1-2 was streaked on a BHI plate, and after culturing at 37°C for 24 hours, a single colony was picked to inoculate liquid BHI medium, cultured with shaking at 37°C until the logarithmic growth phase, and the cells were harvested by centrifugation, and phosphate The buffer solution was washed and suspended, and the bacterial concentration was adjusted to 1×10 10 CFU / ml.

[0023] Irradiation inactivation: Freshly prepared bacterial samples are frozen to 60 The Co radiation source is irradiated, the radiation dose is 15KGy, the radiation method is static radiation, and the dose rate is 60Gy / min.

[0024] Bacterial morphology observed by scanning electron microscope: The radiation inactivated yzu LM 1-2 was observed by scanning electron microscope, the results are as follows figure 1 As shown, no obvious structural damage was seen on the bacterial surface.

Embodiment 2

[0025] Example 2: Immune Effect Evaluation of Irradiated Inactivated Vaccines

[0026] Animal grouping and immunization: Female BALB / c mice aged 6-8 weeks were randomly divided into 5 groups, namely live bacteria group, radiation inactivation group, heat inactivation group, formaldehyde inactivation group and negative control group. Group 24 mice. Live bacteria yzu LM 1-2, radiation-inactivated yzu LM 1-2, heat-inactivated yzu LM 1-2, formaldehyde-inactivated yzu LM1-2 and sterilized PBS were injected intraperitoneally on the 0th and 7th days respectively . The immune dose was 1×10 3 CFU / piece, 1×10 9 CFU / piece, 1×10 9 CFU / piece, 1×10 9 CFU / only and 0.1ml / only.

[0027] Determination of antibody titer in mouse serum after immunization: mice in each group were 7 days after the first immunization, 14 days and 21 days after the second immunization, blood was collected from the orbital vein, and the serum was prepared by centrifugation. The optimal antigen coating concentra...

Embodiment 3

[0035] Embodiment 3: It is basically the same as Embodiment 1, except that the strain used is Listeria monocytogenes LM-4. The obtained radiation inactivated vaccine immune effect evaluation effect is basically the same as that of Example 2.

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Abstract

The invention relates to a Listeria monocytogenes inactivated vaccine and the preparation method, belonging to biotechnical field. The preparation method of Listeria monocytogenes inactivated vaccine is characterized in that Listeria monocytogenes is processed by 60Co irradiation and inactivation to prepare the inactivated vaccine. The protection antigen component in the pathogen surface is remained, which can excite the protection T cell immune response and body fluid immune response. The advantages are significantly important when the inactivated vaccine aiming at intracellular pathogen is researched.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an inactivated Listeria monocytogenes vaccine and a preparation method thereof. technical background [0002] Listeria monocytogenes (LM, also known as Listeria monocytogenes) is an intracellular, facultatively anaerobic, Gram-positive bacillus that is an important food source. A sexually pathogenic bacterium that can cause listeriosis in humans and animals. LM is pathogenic to a variety of livestock and poultry, and has a high mortality rate. The susceptible groups in the population are pregnant women, infants, the elderly and immunocompromised persons, mainly causing meningitis, abortion, and immunodeficiency patients and newborns. Infection of the central nervous system has a low incidence rate, but the mortality rate is as high as 30% to 70%. Therefore, the research on LM has important public health significance. In view of this, people are developing safe and effective vaccine...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61P31/04
Inventor 焦新安董慧潘志明孙林殷月兰
Owner YANGZHOU UNIV