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Function of CH25H and 25-OHC in immune response and application thereof as immunomodulator in vaccines

An immunomodulator and vaccine technology, applied in the field of biomedicine, to improve the immune effect and reduce the effect of cell proliferation and activation

Inactive Publication Date: 2015-10-28
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, two recent clinical trials of HIV vaccines (STEP and HVTN-503) failed for reasons that have yet to be fully explored, but data suggest that these vaccines produced highly proliferative HIV / SIV Gag / Tat-specific CD4+ T cells may be involved in the failure of this vaccine

Method used

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  • Function of CH25H and 25-OHC in immune response and application thereof as immunomodulator in vaccines
  • Function of CH25H and 25-OHC in immune response and application thereof as immunomodulator in vaccines
  • Function of CH25H and 25-OHC in immune response and application thereof as immunomodulator in vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Application of 25-OHC in anti-inflammatory

[0045] The experimental materials used in this embodiment are as follows:

[0046] Cell line: 174×CEM is preserved by our laboratory. It is a fusion cell line of human B lymphocyte line and T lymphocyte line, which is very suitable for the infection experiment of SIV virus. The culture condition of 174×CEM was RPMI 1640 medium containing 10% FBS.

[0047] Chinese macaque PBMCs and mouse lymphocytes: PBMCs were isolated from the whole blood of macaques, and the macaques were raised in the China South African Human Primate Center. Normal mouse spleen lymphocytes were derived from experimental SPF grade C57BL / 6 mice, which were raised in the animal center of Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences. The culture conditions of PBMC and lymphocytes were RPMI 1640 complete medium containing 10% FBS.

[0048] Reagents: OptiPrep™ (Axis-Shield), the main human lymphocyte separation mediu...

Embodiment 2

[0079] Example 2 Application of 25-OHC in Inhibiting Nonspecific Cell Proliferation of Primary Lymphocytes

[0080] method:

[0081] A. Stain mouse spleen single cell suspension or rhesus monkey PBMC according to conventional CFSE experiments

[0082] 1) Separation of lymphocytes by conventional methods, trypan blue staining and cell counting, with 4×10 6 The number of cells (4 wells for each stimulating peptide, 1 million cells / well, 24 wells) is a staining unit. Transfer to a 15 ml centrifuge tube and resuspend with an appropriate volume of RPMI 1640. After centrifugation at 350 g for 5 min, resuspend in 1 ml RPMI 1640.

[0083] 2) Dilute the CFSE stock solution with RPMI1640 to below 2 μmol / L (2 mM CFSE stock solution is diluted 1000 times) and operate in the dark as much as possible.

[0084] 3) Add 1 ml of CFSE diluent to every 1 ml of cell suspension, and mix gently (at this time, the cells are fragile, so the action should be as gentle as possible).

[0085] ...

Embodiment 3

[0102] Example 3: Modulation of vaccine immunogenicity by 25-OHC

[0103] Studies have shown that in vitro cell experiments have shown that excessively high concentrations of 25-OHC often have obvious toxic effects on cells, resulting in cell apoptosis and weakened proliferation. However, no literature has been found to study the in vivo toxicity of 25-OHC exceeding physiological concentrations. The plasma of normal mice contains only a very low concentration of 25-OHC (about 2 ng / ml). In this study, we used continuous tail vein injection of low-dose and high-dose 25-OHC to observe its effect on the immunogenicity of SIV antigen.

[0104] The experimental materials used in this embodiment are as follows:

[0105] Experimental animals: SPF grade C57BL / 6 mice (Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.), raised by the Animal Experiment Center of Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences.

[0106] SIV vaccine (Ad5-SIVenv):...

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Abstract

The invention discloses the function of CH25H (cholesterol-25-hydroxylase) and 25-OHC in immune response and application thereof as an immunomodulator in vaccines. The invention finds that 25-OHC can significantly inhibit inflammation immunologic reaction, can inhibit immune cell overexpressed IL-1b and TNF-a, and has very important application value for adjustment of virus infection patient persistent immune activation and inflammatory reaction. Without affecting antigen specific T lymphocyte proliferation, 25-OHC strongly inhibits mitogen stimulated lymphocyte overproliferation effect. 25-OHC can be used as the immunomodulator of a HIV (human immunodeficiency virus) vaccine to improve the immune effect of the vaccine, and can be widely applied to prevention and treatment of AIDS. The mentioned immunomodulatory effect is in favor of designing a new generation of HIV vaccine, i.e. reducing CD4<+> T cell immune response to the greatest extent without affecting body protective CD8<+> T cell immune functions.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the role of a gene and its coding and metabolites in regulating immune response, and the application of the immune regulating effect in developing new AIDS drugs and vaccines. Background technique [0002] Cholesterol-25-hydroxylase (cholesterol-25-hydroxylase, CH25H) and its catalyzed metabolite 25-OHC (25-hydroxycholesterol) are closely related to lipid metabolism, for example, 25-OHC can inhibit the synthesis of sterols. Recent studies have shown that CH25H is a member of the interferon-stimulated gene family (ISG gene). Studies have shown that CH25H and its metabolite 25-OHC have broad-spectrum antiviral activity. However, its exact antiviral mechanism and its role in regulating immune responses, especially The regulation of antigen-specific immune responses is unclear. [0003] Human acquired immunodeficiency virus (Human Immunodeficiency Virus, HIV) and AIDS (AIDS) caused by it ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/44A61K31/575A61K39/21A61P37/02A61P31/18
Inventor 孙彩军陈凌吴同金
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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