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Compound immunological adjuvant and vaccine

An immunoadjuvant and compound technology, applied in antiviral agents, pharmaceutical formulations, viral antigen components, etc., can solve the problem of unsatisfactory adjuvant effect, achieve obvious adjuvant effect, improve cellular immune response, and improve humoral immune response. Effect

Active Publication Date: 2012-05-30
国家兽用生物制品工程技术研究中心 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the adjuvants in the current prior art often only stimulate Th1 or Th2 immune responses alone, and the comprehensive adjuvant effect is not ideal enough, so the research of this invention is especially carried out

Method used

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  • Compound immunological adjuvant and vaccine
  • Compound immunological adjuvant and vaccine
  • Compound immunological adjuvant and vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Preparation of vaccine using gonadotropin-releasing hormone (abbreviated as GnRH) synthetic peptide as immunogen

[0018] The antigen is GnRH synthetic peptide (for castration of animals), which was synthesized by Gill Biochemical (Shanghai) Co., Ltd. The synthetic peptide sequence is: Th-GG-QHWSYGLRPGQHWSYGLRPGQHWSYGRPGQHWSYGLRPG.

[0019] Imiquimod was purchased from Wuhan Risheng Technology Development Co., Ltd.

[0020] The preparation process of compound immune adjuvant and vaccine is as follows:

[0021] (1) Preparation of CpG aqueous phase solution: prepare CpG into a solution with a concentration of 2.0 mg / mL with double distilled water, add Tween-80 until the volume concentration reaches 4%, filter and sterilize through a 0.22 μm filter membrane to obtain the aqueous phase solution;

[0022] (2) Preparation of imiquimod oil phase solution: add 1.0 mg of imiquimod to each mL of white oil to prepare an imiquimod white oil solution with a concentratio...

Embodiment 2

[0026] Example 2 Immunization test on mice using GnRH synthetic peptide vaccine as immunogen

[0027] Referring to Example 1, at the same time, under the conditions of constant antigen dose and volume (insufficient part is supplemented with normal saline), respectively prepare imiquimod+white oil, CpG+white oil, single white oil as adjuvant Three vaccine control groups.

[0028] The vaccines prepared in Example 1 and the three control groups were used for group immunization respectively.

[0029] Immunization method: Male mice (18-20 g) aged 3-4 weeks were randomly divided into 4 groups, 10 mice in each group. The immunization dose is 200 μl / only. The immunization method is as follows: first immunization at the age of 28 days, and then booster immunization with the same dose at the age of 56 days.

[0030] Sampling and processing: Blood was collected once a week after immunization, and the blood sample was first placed at 37°C for 2 hours, then overnight at 4°C, centrifuged...

Embodiment 3

[0037]Example 3 Immune Test of Vaccine Using Newcastle Disease IV Inactivated Antigen as Immunogen

[0038] Using the same method as in Example 1, prepare a vaccine with Newcastle disease IV inactivated antigen, the difference is that the prepared CpG concentration in the aqueous phase solution is 1.0 mg / mL, and the imiquimod concentration in the oil phase solution is 0.5 mg / mL, the antigen adopts chicken Newcastle disease IV inactivated antigen (the virus titer before inactivation is 10 8.5 EID 50 / 0.1mL). Using inactivated Newcastle disease Ⅳ antigen as the immunogen, four groups of vaccines were prepared with the following adjuvants: imiquimod+CpG+white oil, imiquimod+white oil, CpG+white oil, and white oil. The 28-day-old chicks were immunized with four groups of vaccines (the titer of Newcastle disease maternal antibody was <2.0 Log 2), blood was collected at 2 weeks, 3 weeks and 4 weeks after immunization, the serum was separated, and the hemagglutination inhibition (H...

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Abstract

The invention relates to a compound immunological adjuvant comprising a water phase solution and an oil phase solution which are respectively prepared, wherein the water phase solution is a CpG water solution the concentration of which is 0.5 to 2.0mg / mL; the oil phase solution is an imiquimod white oil solution the concentration of which is 0.25 to 1.0mg / mL; and the volume ratio of the water phase solution to the oil phase solution is 1:6. The invention also relates to a vaccine prepared from one or more antigens selected from attenuated live full microorganisms, inactivated microorganisms, recombinant peptide and proteins, synthetic peptide and cracked microorganisms and the water phase solution and the oil phase solution in the compound immunological adjuvant according to the volume ratio of 1:1:6. By the compounding of imiquimod, CpG and a white oil component, the compound immunological adjuvant has an obvious synergistic effect and enhances the immunological activity reactions ofTh1 and Th2, so that the activity of stimulated immunological cells is obviously enhanced.

Description

technical field [0001] The invention relates to a compound immune adjuvant and a vaccine containing the immune adjuvant. Background technique [0002] When the antigen is immunized alone, it is not enough to produce immunogenicity, especially the highly pure antigen (such as polypeptide). Therefore, different kinds of adjuvants are required to enhance the immune response induced by the antigen. The addition of adjuvants can reduce the number of antigens in the vaccine or the number of boosters or prolong the protection time of the vaccine. [0003] Toll-like receptors (Toll-like receptors, TLRs) are a new class of innate immune receptors on the cell surface discovered in recent years. immune cells, eventually causing a series of inflammatory responses. TLRs can not only activate the innate immune system, but also provide co-stimulatory signals for the activation of acquired immunity. TLRs receptor agonists can maximize the activation of dendritic cells, improve the abili...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/00A61K39/17A61P31/14
Inventor 卢宇张金秋邓碧华侯继波
Owner 国家兽用生物制品工程技术研究中心
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