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Immunomicrosphere for overcoming B cell immunological tolerance and application thereof

A technology of immune microspheres and immune tolerance, which is applied in the field of immunity and can solve problems such as immune competition

Inactive Publication Date: 2011-04-13
SHANGHAI WEIQIU BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0019] In order to solve the above problems, the present invention provides a kind of immune microspheres, so as to overcome the immune competition problem of chemically cross-linked protein or fusion protein molecular epitope in the prior art

Method used

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  • Immunomicrosphere for overcoming B cell immunological tolerance and application thereof
  • Immunomicrosphere for overcoming B cell immunological tolerance and application thereof
  • Immunomicrosphere for overcoming B cell immunological tolerance and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Preparation of Calcium Diatomate / Chitosan Microspheres Encapsulating Bovine Serum Albumin in Example 1

[0057] 1. Preparation of calcium diatomate microspheres coated with bovine serum albumin (BSA)

[0058] Dissolve 2 g of sodium alginate in 90 ml of double distilled water, and add bovine serum albumin (BSA) to a final concentration of 1 mg / ml. Adjust the pH to 7.3 with 0.5M NaOH, add water to make up to 100ml to prepare a 2% (m / v) solution. Calcium chloride (CaCl2) was prepared as a 10% (m / v) solution. 2% sodium alginate solution 4ml air pump spray pen atomization spray into 100ml calcium chloride solution, 500 rpm magnetic stirring to form a ball. Equilibrate with magnetic stirring for 1 hour, collect microspheres by centrifugation, stir and balance with double distilled water for 1 hour, and wash by centrifugation for 3 times.

[0059] The size and uniformity of the microspheres were checked by microscopy. Judging from the comparison of the size of red blood ce...

example 2

[0075] Example 2, preparation of polylactic acid-glycolic acid (PLGA) / chitosan microspheres wrapped with ovalbumin (OVA)

[0076] For the preparation of microspheres used in this example, reference can be made to: (1) Preparation, Characterization and In vitro Sustained Release of Drug-loaded Microspheres; Postgraduate Thesis of Tsinghua University (2008) Wang Yiwei, Cui Fuzhai. (2) Preparation of polylactic acid microspheres; Journal of Jilin University (Science Edition) Vol.43N o.6Nov 2005: 842-846.

[0077] 1. Polylactic acid-glycolic acid copolymer was purchased from SIGMA Company (the composition ratio was 50:50), and 100 mg was dissolved in 5 mL of organic solvent dichloromethane. Weigh 5 mg of OVA, and grind the OVA particles finely with a bacterial grinder. Suspended in an organic solvent containing PLGA. And slowly pour it into 50mL of water phase containing 1% polyvinyl alcohol (PVA), and ultrasonic emulsify with 300W power for 30s under ice bath. Then the double ...

example 3

[0084] The agarose microsphere preparation of example 3 wrapping BSA

[0085]1. Preparation method: Low solidification temperature agarose was purchased from SIGMA Company. Agarose was melted with distilled water at a concentration of 4%. Put it in a 45C degree water bath. BSA was dissolved in 0.05M pH7.2 phosphate buffer at a concentration of 0.2%. Equal volume of protein solution was mixed with agarose to make the final concentration: agarose was 2%, protein concentration was 0.5%.

[0086] The microspheres are prepared by the membrane emulsion method, and the specific operation can refer to the literature: Preparation of PLGA microspheres and microcapsules with uniform particle size by rapid membrane emulsification method. Tian Rui, Wang Lianyan et al. Journal of Process Engineering Vol.9No.4. August 2009. 754-761.

[0087] The preparation of microspheres by the membrane emulsification method is to disperse the melted agarose into the oil phase without passing through...

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Abstract

The invention relates to an immunomicrosphere for overcoming B cell immunological tolerance and application thereof. The immunomicrosphere comprises a microsphere medium, wherein the outside of the microsphere medium is coated with vaccine antigen, and the inside of the microsphere medium is coated with immune carrier protein for activating T cells. The invention also relates to application of the immunomicrosphere for overcoming the B cell immunological tolerance in preparing vaccines and immunological adjuvants. By using the technical scheme, the immunomicrosphere is combined with B cell receptor through the antigen and phagocytized by B cells, and the carrier protein is released and degraded in the cells; the immunomicrosphere is presented to Th cells through B cell MHC II to activate the TH cells to release cell factors, thereby promoting the activation and proliferation of the B cells; and high-titer antibody is generated through the affinity maturation process. The antigen protein and the immune carrier protein are respectively arranged on the surface of the microsphere and inside the microsphere, thereby preventing the antigen protein epitope and the Th cell activation epitope from competing with the B cell receptor.

Description

technical field [0001] The invention relates to the field of immunity, in particular to an immune microsphere for overcoming B cell immune tolerance and application thereof. Background technique [0002] Antigen (Ag) refers to a substance that can bind to TCR or BCR of T and B lymphocytes, promote their proliferation and differentiation, produce antibodies or sensitize lymphocytes, and then exert immune effects. There are many ways to classify antigens. According to whether Th cells need to participate in the induction of antibodies, they can be divided into thymus-dependent antigens (TD-Ag) and thymus-independent antigens (TI-Ag). The vast majority of protein antigens, such as pathogenic microorganisms, blood cells, and serum proteins, belong to TD-Ag. Thymus-dependent antigen-induced antibody production process requires the participation of T cells. The instructions for antibody production come from two signals: antigen binding to B cell surface antigen receptors and co-...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/39A61K9/16A61K47/48A61P35/00A61K47/68
Inventor 谈立松张尚权陈宇光
Owner SHANGHAI WEIQIU BIOTECH
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