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Bacterial toxin vaccine and application thereof in preventing bacterial infection

A technology of bacterial toxins and vaccines, which is applied in the field of related and genus health, and can solve problems such as the difficulty in maintaining the activity of red blood cell membranes, the complicated process of coating nanoparticles on cell membranes, and the difficulty in large-scale production.

Inactive Publication Date: 2021-06-29
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the nano-toxoid vaccine still has some obvious deficiencies, such as low toxin load (taking α-hemolysin as an example, only 22 μg / mg), weak antigen presentation effect, immune activation ability needs to be improved, and cell membrane The process of coating nanoparticles is relatively complicated, the activity of red blood cell membrane is difficult to maintain, the resources of red blood cells are limited, large-scale production is difficult, and clinical transformation is greatly restricted

Method used

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  • Bacterial toxin vaccine and application thereof in preventing bacterial infection
  • Bacterial toxin vaccine and application thereof in preventing bacterial infection
  • Bacterial toxin vaccine and application thereof in preventing bacterial infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: the construction of liposome vaccine

[0037] In this example, high cholesterol liposomes were first prepared by film hydration extrusion method. Specific operation: Weigh cholesterol, phosphatidylcholine (PC), DSPE-PEG2000 and / or sphingomyelin (as shown in Table 1) and dissolve them in dichloromethane, and spin evaporate at room temperature to form a lipid film. Add an appropriate amount of deionized water and stir at room temperature for 1 hour of hydration, 100W ultrasonic to obtain liposome (PLs) suspension. The toxin load of PLs was determined by in vitro hemolysis assay. Specific operation: Incubate 3 μg Hlα with different amounts of PLs at room temperature for 30 minutes, then incubate with appropriate amount of 2.5% RBC at 37°C for 3 hours, centrifuge at 2000 g for 5 minutes, measure the absorbance value of the supernatant at 540 nm and calculate the percentage of hemolysis according to the following formula. The negative control is physiological...

Embodiment 2

[0041] Embodiment 2: Physicochemical property characterization of liposome vaccine

[0042] The high-cholesterol liposomes (PLs) without toxin were used as the control. After negative staining with uranyl acetate, the shape and size of PLs(Hlα) were observed by transmission electron microscopy, and the results showed ( figure 1 B), Fused liposomes are regular spherical and uniform in size, similar to PLs. The average particle size, particle size distribution and polydispersity coefficient of PLs (Hlα) were measured by dynamic light scattering method, and its Zeta potential was measured. The results showed that the average particle size of PLs (Hlα) was 125nm, and the particle size distribution was narrow, and the potential was -35mV, which was similar to PLs ( figure 1 C, D). The above results indicated that the prescription amount of toxin loading had no significant effect on the physicochemical properties of fusion liposomes.

Embodiment 3

[0043] Example 3: Safety Evaluation of Liposome Toxoid Vaccine

[0044] In this example, the in vitro safety of PLs (H1α) was first investigated through hemolysis experiments, and compared with H1α and heat-inactivated H1α. Specific operation: Hlα, heat-inactivated Hlα (70°C for 30min or 60min), PLs, PLs(Hlα) were shaken and incubated with 2.5% RBC for 3 hours at 37°C, then centrifuged at 2000g for 5min, and the absorbance of the supernatant at 540nm was measured Value and calculate the percentage of hemolysis according to the formula of Example 1. the result shows( figure 2 A), Hlα inactivation at 70°C for 30 minutes still has a certain hemolysis effect, and inactivation at 70°C for 60 minutes can completely eliminate the hemolysis effect of Hlα, while PLs (Hlα) has no hemolysis effect, showing better safety.

[0045] Next, the in vitro cytotoxicity of PLs (Hlα) was investigated. Specific operation: Extract mouse bone marrow and culture to obtain primary dendritic cells. ...

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Abstract

The invention belongs to the technical field of biology, and relates to a bacterial toxin vaccine, and particularly relates to a liposome vaccine of high-load bacterial pore-forming toxin and application thereof. The nano vaccine completes high-efficiency load of the toxin by embedding the bacterial pore-forming toxin into the surface of liposome, so that the toxic effect of the toxin is eliminated, and the antigen protein structure is completely reserved; and after inoculation, high-efficiency presentation of immune cells on the pore-forming toxin can be caused, a high-titer specific antibody is generated, and bacterial infection, particularly drug-resistant bacterial infection is effectively prevented.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a bacterial toxin vaccine, in particular to a liposome vaccine loaded with a bacterial pore-forming toxin and its application. The liposome vaccine embeds the bacterial pore-forming toxin into an artificial liposome bilayer membrane Completing the toxin load not only eliminates the toxic effect of the toxin, which has high safety, but also produces an efficient immune response after vaccination, which can effectively prevent bacterial infections, especially drug-resistant bacterial infections. Background technique [0002] According to reports, bacterial infection is still a class of diseases with high morbidity and high fatality rate worldwide, and the emergence of drug-resistant bacteria, especially "super bacteria", has brought greater challenges to the treatment of anti-bacterial infections, seriously threatening human health. At present, there is a lack of research and de...

Claims

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Application Information

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IPC IPC(8): A61K39/085A61K9/127A61K47/24A61K47/28A61P31/04A61P31/00A61P33/00A61P39/02A61P35/00
CPCA61K39/085A61K9/1271A61K47/24A61K47/28A61P31/04A61P31/00A61P33/00A61P39/02A61P35/00
Inventor 庞志清刘春影
Owner FUDAN UNIV
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