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Aphanothece halophytica expolysaccharide immunoadjuvant

A technology of cryptobacterium algae and immune adjuvant, applied in the field of biomedicine, can solve the problems of inability to induce immunity, difficult standardization of quality standards, swelling, erythema, pain, induration and ecchymosis, etc.

Inactive Publication Date: 2013-08-28
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Aluminum adjuvant is the only adjuvant approved by the FDA so far that can be used in humans, but it has some unavoidable shortcomings: (1) Studies have confirmed that the injection site of the human body after the aluminum adjuvant (0.6mg / ml) is mixed with the antigen There are adverse reactions such as swelling, erythema, pain, induration and ecchymosis; (2) in the process of preparing aluminum glue, the quality standard is difficult to standardize, and the problem of long-term storage has not been well resolved; (3) aluminum glue Adjuvants can only induce humoral immunity, but not cell-mediated immunity, which is especially important for the body. Aluminum adjuvants are far from meeting the development requirements of new vaccines
The exopolysaccharide of C. salina described in the present invention has not been reported as an immune adjuvant

Method used

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  • Aphanothece halophytica expolysaccharide immunoadjuvant
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  • Aphanothece halophytica expolysaccharide immunoadjuvant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Hemolytic determination

[0014] In the experiment, rabbit heart was used to take blood, first inhale 3ml of sterile Ahrlid solution in the syringe, then take 7ml of blood from the heart, quickly push it into 4 times the volume of normal saline, shake well, centrifuge, and repeat the above operation with normal saline 3 The second highest clear was light yellow. Add 20ml of Aldrin's solution and mix and store at 4°C for later use.

[0015] The above red blood cells were diluted with normal saline into a 2% red blood cell suspension for experiment. The extracellular polysaccharide of Cryptococcum salina was diluted to 0.25, 0.50, 1.00 mg / ml with normal saline. Take 2ml of each and add 2ml 2% red blood cell suspension, mix well, and set the maximum and minimum hemolytic control with distilled water and normal saline respectively The above groups were placed in a constant temperature water bath at 37°C for 1 hour, the experimental phenomenon was observed, centrifuged, the sup...

Embodiment 2

[0020] In vitro lymphocyte transformation experiment

[0021] Take fractional dislocations and put them to death. Take the spleen aseptically, add PBS solution, grind with a tissue homogenizer, and filter with a 100-mesh sieve to prepare a single cell suspension. After centrifugation, the supernatant was discarded, washed twice with PBS solution, and red blood cell lysate Tris-NH was added 4 Cl(NH 4 Cl 7.47g, Tris 2.60g, add ddH 2 0 to 1L, sterilized by 0.22 membrane filtration after autoclaving) 5ml, mix well and place at 37°C for 5min, add an equal volume of RPMI1640 complete medium (containing 10% FBS, 100μg / ml streptomycin, 100μg / ml penicillin) Discard the supernatant after centrifugation in the same way as above. Then wash twice with 1640 medium without serum, suspend the cells in 1640 complete medium, dilute with 10μl of spleen cell suspension, stain with trypan blue and count, the number of viable cells is not less than 95%; then prepare 5×10 6 A / ml mouse spleen lymphocyte...

Embodiment 3

[0025] The effect of Cryptococcum salina exopolysaccharide on the cellular immunity of OVA-immunized mice

[0026] In this example, the physiological saline solution of C. salina exopolysaccharide and the physiological saline solution of OVA were uniformly mixed to obtain a mixed solution of C. salina exopolysaccharide and OVA; the physiological saline solution of aluminum gum adjuvant It is evenly mixed with the physiological saline solution of OVA to obtain a mixed solution of aluminum gel adjuvant and OVA.

[0027] 40 mice were randomly divided into groups, 8 mice in each group. Normal saline negative control group: each mouse was injected with 0.2ml normal saline; OVA control group: each mouse was injected with 0.2ml OVA physiological saline solution (0.5mg / ml); aluminum gel adjuvant (ALum) control group: each Mice were injected with a mixture of 0.1ml OVA physiological saline solution (1mg / ml) and 0.1ml aluminum gel adjuvant (ALum) physiological saline solution (2mg / ml); Cryp...

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Abstract

The present invention relates to an application of aphanothece halophytica expolysaccharide in immunoadjuvant preparation. With application of aphanothece halophytica expolysaccharide as an immunoadjuvant, immune response reactions of a human body immune system to immunogen can be significantly increased, wherein the immune response reactions comprise body fluid immune response and cell immune response, the body fluid immune response effect is the same as an effect of an alhydrogel adjuvant while a side effect of the alhydrogel adjuvant does not exist, and the alhydrogel adjuvant can not stimulate a cell immune response reaction, such that the aphanothece halophytica expolysaccharide immunoadjuvant is superior to the alhydrogel adjuvant only approved by the FDA and used for human.

Description

Technical field [0001] The present invention relates to the field of biomedicine, in particular to the use of Cryptocoryne salina extracellular polysaccharide as an immune adjuvant. Background technique [0002] Immune adjuvants are a class of substances that can improve the immunogenicity of antigens and improve the effectiveness of vaccines. It is the adjuvant technology that gives vaccines a clear advantage in price. At present, the most commonly used adjuvants are aluminum gel adjuvant and Freund's adjuvant. Aluminum gel adjuvant is the only adjuvant approved by the FDA for use in humans, but it has some uncontrollable shortcomings: (1) Studies have confirmed that the aluminum adjuvant (0.6mg / ml) is mixed with the antigen for injection in humans. There are adverse reactions such as swelling, erythema, pain, induration and ecchymosis; (2) The quality standards are difficult to standardize during the preparation process of aluminum glue, and the problem of long-term storage ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61P37/04
Inventor 欧瑜韦庆芳
Owner CHINA PHARM UNIV
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