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Application of polyinosinic-polycytidylic acid (poly I:C) combined dimo-thylidioctyl ammonium bromide (DDA) mixed adjuvant in preparation of tuberculosis subunit vaccines

A dimethylhexadecane and subunit vaccine technology, which is applied to medical preparations containing active ingredients, bacterial antigen components, antibody medical components, etc., can solve the problems of difference in effectiveness and poor protection effect, and achieve relief Pathological damage, enhance secretion level, reduce the effect of lung pathological damage

Active Publication Date: 2013-06-05
LANZHOU UNIVERSITY
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Problems solved by technology

For decades, BCG vaccine has been widely used in the world, but its effectiveness varies greatly in different regions
BCG was also less protective in adults

Method used

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  • Application of polyinosinic-polycytidylic acid (poly I:C) combined dimo-thylidioctyl ammonium bromide (DDA) mixed adjuvant in preparation of tuberculosis subunit vaccines
  • Application of polyinosinic-polycytidylic acid (poly I:C) combined dimo-thylidioctyl ammonium bromide (DDA) mixed adjuvant in preparation of tuberculosis subunit vaccines
  • Application of polyinosinic-polycytidylic acid (poly I:C) combined dimo-thylidioctyl ammonium bromide (DDA) mixed adjuvant in preparation of tuberculosis subunit vaccines

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Experimental program
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Embodiment 1

[0026] Dissolve PolyI:C in phosphate buffer to 0.5 mg / ml, dilute the fusion protein AMM to 0.4 mg / ml with sterile PBS, prepare DDA with sterile distilled water to a 2.5 mg / ml solution, bathe in 80°C for 10 minutes, and cool to room temperature; mix 50 μl of PolyI:C solution with an equal amount of AMM protein solution, and place at room temperature for 1 min; add 100??L DDA solution to the mixed solution drop by drop, and then fully emulsify the vaccine to obtain a uniform creamy oil. Subunit vaccine AMM+DDA / PolyI:C.

[0027] The mice in each experimental group were subcutaneously immunized in the groin, and the total volume of the vaccine was 200ul.

[0028] Two vaccine immunization strategies were adopted, one was booster immunization with fusion protein vaccine; the other was booster immunization with fusion protein vaccine after BCG primary immunization.

[0029] 1. Fusion protein AMM vaccine group with DDA / PolyI:C as adjuvant (AMM+DDA / PolyI:C), fusion protein AMM vaccine...

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Abstract

The invention discloses application of a polyinosinic-polycytidylic acid (poly I:C) combined dimo-thylidioctyl ammonium bromide (DDA) mixed adjuvant to the preparation of tuberculosis subunit vaccines. The invention has the advantages that: the mixed adjuvant DDA / Poly I:C can enhance the secretion level of antigenic specificity interferon-gamma (IFN-gamma) of fusion protein Ag85B-MPT64190-198-Mtb8.4 (AMM) effectively and relieve pathological damage caused by H37Rv infection, and is an effective adjuvant of vaccines for preventing tuberculosis.

Description

technical field [0001] The invention relates to the field of biological preparations, in particular to the application of a mixed adjuvant of polymuscular cell and dimethyl hexadecyl ammonium in the preparation of tuberculosis subunit vaccine. Background technique [0002] Tuberculosis is one of the major infectious diseases causing human death worldwide. Mycobacterium bovis Bacilli Calmette Guerin (BCG) is currently the only vaccine against tuberculosis. BCG is derived from the attenuated strain of Mycobacterium bovis in the last century, and it has been subcultured in vitro for 13 years. For decades, BCG vaccine has been widely used in the world, but there are huge differences in its effectiveness in different regions. BCG was also less protective in adults. Therefore, a safe and effective anti-TB vaccine is essential. Among the many new vaccines, subunit protein vaccines are the most promising vaccines. [0003] Due to the weak immunogenicity of subunit vaccines, app...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/04A61P31/06
Inventor 祝秉东达泽蛟牛红霞辛奇于红娟李青章国平李志刘万波张颖
Owner LANZHOU UNIVERSITY
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