Application of porphyrin pigment serving as immunologic adjuvant and vaccine

An immune adjuvant and pigment technology, applied in the field of immunology, can solve the problems of limited enhancement effect and prolonged stimulation, and achieve the effect of clear metabolic pathway

Inactive Publication Date: 2015-05-20
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But Al(OH) 3 The adjuvant itself has no stimulating effect on the immune system. It mainly prolongs the stimulation of antigens to lymphoid tissue through the depot effect of adsorption, storage and slow release of antigens, and the enhancement effect is limited.

Method used

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  • Application of porphyrin pigment serving as immunologic adjuvant and vaccine
  • Application of porphyrin pigment serving as immunologic adjuvant and vaccine
  • Application of porphyrin pigment serving as immunologic adjuvant and vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The immunoenhancing effect of the combination of iron porphyrin, lecithin and iron porphyrin on the weak antigen bovine serum albumin was tested.

[0045] Iron porphyrin (FePP) : Also known as heme, the molecular formula is C 34 h 32 FeN 4 o 4 , the molecular mass is 616.49, the structure is as attached figure 2 Shown; Adopt the following method to prepare and obtain in the present embodiment, of course, also can adopt commercially available product.

[0046] (1) Collect animal or human anticoagulant blood, centrifuge at 4000 rpm, collect red blood cells, and wash red blood cells three times with 0.1M phosphate buffered saline (PBS, pH7.4).

[0047] (2) Add an equal volume of deionized water to the washed red blood cells, add 5 times the volume of chloroform after complete hemolysis and mix well, let it stand for about 10 minutes, and a cake-shaped hemoglobin precipitate will form on the upper layer of the 50mL centrifuge tube (the lower layer is clear chloroform...

Embodiment 2

[0089] The immunoenhancing effect of the combination of cobalt porphyrin, lecithin and cobalt porphyrin on the weak antigen bovine serum albumin was tested.

[0090] Cobalt Porphyrin (CoPP) : Molecular formula is C 34 h 32 CoN 4 o 4 , the molecular mass is 619.58, the structure is as attached image 3 As shown, the CoPP in this example was purchased from Sigma Company with a purity of ≥98%.

[0091] the solution

[0092] BSA powder (purchased from Sigma, purity ≥98%) was dissolved in 0.1 M phosphate buffer (PBS, pH 7.4) to prepare a 15 μM (1 mg / mL) BSA solution, which was stored in a refrigerator at 4 °C for use.

[0093] the solution

[0094] First dissolve the CoPP powder with a small amount of 0.1M NaOH, then add PBS to make a 2mM (1.239mg / mL) CoPP solution, and store it in a 4°C refrigerator for later use.

[0095] the solution

[0096] The 15 μM BSA solution was mixed with an equal volume of 2 mM CoPP solution, so that the final concentrations of BSA and CoP...

Embodiment 3

[0118] To test the immune effect of hepatitis B vaccine containing iron porphyrin pigment on BALB / c mice.

[0119] Iron porphyrin (FePP)

[0120] Prepared by the same method as in Example 1, the purity is greater than or equal to 95%. Of course, commercially available products can also be used.

[0121] (HBsAg) solution

[0122] The product of Shanghai Yemin Biotechnology Co., Ltd., the purity is ≥95%, and the concentration is 2.5mg / mL.

[0123] FePP solution

[0124] Dissolve the FePP powder with a small amount of 0.1M NaOH first, then add PBS to make a 2mM (1.233mg / mL) FePP solution, and store it in a 4°C refrigerator for later use.

[0125] the solution

[0126] Dilute an appropriate amount of HBsAg stock solution 5 times and mix it with an equal volume of 2mM FePP solution. The final concentrations of HBsAg and FePP are 0.25mg / mL and 1mM, respectively, and store in a refrigerator at 4°C for later use.

[0127] the solution

[0128] (1) Add 30mg of lecithin ...

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Abstract

The invention discloses application of a porphyrin pigment serving as immunologic adjuvant and further discloses application of the porphyrin pigment serving as a vaccine. The porphyrin pigment is a biological pigment, tissues outside an animal cardiovascular system can be combined with a pathogeny recognition receptor TLR4, and tissue damage and danger signals are simulated to build efficient stimulation to an immune system. The porphyrin pigment is also an ingredient of an animal and a human body, internal metabolic pathways are clear, uncertainty in the safety aspect does not exist, and the porphyrin pigment is suitable for preparation of the immunologic adjuvant and the vaccine for the animal, especially for human. The porphyrin pigment serving as the immunologic adjuvant can remarkably strengthen the humoral immunity effect of antigen or the vaccine and can reach the level of complete Freund adjuvant.

Description

technical field [0001] The invention belongs to the field of immunology, and specifically relates to the application of porphyrin pigment as immune adjuvant and vaccine. Background technique [0002] The problem in modern immunotherapy and immunoprevention is not the immunogenicity of the vaccine itself, but the safety of vaccine preparations (non-polluting factors). Traditional vaccines are low-virulence or inactivated pathogenic microorganisms, which have good immune effects but are highly dangerous; therefore, modern vaccines usually use a certain non-toxic signature component of pathogenic microorganisms or their artificial simulants as antigens, called subunit vaccines or epitope vaccines. Modern vaccines have greatly weakened the immunogenicity while eliminating the danger, and immune adjuvants are needed to make up for the lack of immunogenicity, which has led to the research of modern vaccines essentially becoming the research of immune adjuvants, and at the same ti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/39A61K39/29A61P37/04A61P31/20
Inventor 李国富刘晗范增薛海艳
Owner SUN YAT SEN UNIV
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