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Exosome of Schistosoma japonicum egg antigen (SEA) bone marrow dendritic cell origin and application thereof

A technology of dendritic cells and schistosomiasis, applied in the field of medicine and biology, can solve the problems of parasitic infection and disease, and achieve the effect of avoiding toxic and side effects

Active Publication Date: 2016-08-31
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Recent studies suggest that the occurrence of immune enteritis may be related to the deviation of T cell subsets. Therefore, correcting the unbalanced immune response from the aspect of immune regulation is an important method for the treatment of immune enteropathy. Parasitic infection can induce host Produce immune tolerance and regulate the deviation of host T cell subsets, which may have the effect of treating immune enteropathy, but the use of live parasites to directly infect patients has medical ethics restrictions, and there is also a risk of parasitic infection causing disease. However, finding and discovering insect-derived molecules with immunomodulatory effects such as schistosomiasis egg antigen (SEA) from parasites, and developing them into drugs can completely avoid this risk

Method used

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  • Exosome of Schistosoma japonicum egg antigen (SEA) bone marrow dendritic cell origin and application thereof
  • Exosome of Schistosoma japonicum egg antigen (SEA) bone marrow dendritic cell origin and application thereof
  • Exosome of Schistosoma japonicum egg antigen (SEA) bone marrow dendritic cell origin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation method of Schistosoma japonicum egg antigen (SEA)

[0027] Dissect the New Zealand rabbits infected with Schistosoma japonicum, take out the liver, remove the large blood vessels, bile ducts and connective tissues, cut them into small pieces, put them in a tissue masher, add an appropriate amount of 4°C pre-cooled normal saline, and mash at 3000 rpm for 3 times, 1 min each time, with an interval of 3 min. The liver homogenate was sieved through 50, 100, 120 and 140 mesh / inch sampling sieves respectively, and the collected filter residues were repeatedly crushed according to the above steps, filtered and combined with the above filtrate. Put the filtrate in a glass sedimentation cup to settle for 1 hour, suck up the upper layer of liver slag suspension with a negative pressure suction device (be careful not to suck up the eggs at the bottom, and leave the golden yellow eggs at the bottom of the tube, wash the sediment several times with water, 45 μm...

Embodiment 2

[0028] Example 2 The method of stimulating DC to produce exosomes by Schistosoma japonicum egg antigen (SEA)

[0029] After the BALB / c mice were sacrificed by cervical dislocation, they were soaked in 75% alcohol for 5 minutes, and the femur and tibia were taken out under aseptic conditions. Wash out the bone marrow cells with serum-free RPMI1640 medium, wash repeatedly until the bone turns white, collect the cell suspension and place it in a 15mL centrifuge tube, centrifuge at 1500rpm for 5min, discard the supernatant, and resuspend the pelleted cells with 5mL serum-free RPMI1640 medium, 1: Add the erythrocyte lysate pre-warmed at 37°C at a volume ratio of 10, pipette repeatedly to mix, and place at room temperature for 2 to 3 minutes to break the erythrocytes. Then centrifuge and wash with PBS twice, collect the pelleted cells and resuspend in 5 mL of complete RPMI-1640 DC culture medium (containing 10 mM Hepes, 2 mM glutamine, 100 U / mL penicillin and 100 U / mL streptomycin, ...

Embodiment 3

[0034] Example 3 The exosome prepared in Example 2 treats the mouse model of inflammatory bowel disease

[0035] Treatment of inflammatory bowel disease with dendritic cell-derived exosome stimulated by Schistosoma japonicum egg antigen (SEA).

[0036] 1. Establishment method of inflammatory bowel disease (IBD) model

[0037] Inflammatory bowel disease (IBD) model was induced in BALB / C mice using dextran sodium sulfate (Dextran sodium sulphate, DSS): DDS was dissolved in direct drinking water to prepare a 3% solution (W / V) , and fed 6-week-old BALB / C mice from the 0th day to the 8th day. The weight change of the mice was recorded every day, the feces were collected, the shape of the feces was observed, and occult blood was measured, and the disease activity index of the mouse model was calculated based on the weight change, occult blood, and feces shape.

[0038] DSS and BALB / C mice were used to construct a mouse model of inflammatory bowel disease, and exosomes derived from...

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Abstract

Schistosoma japonicum egg antigen (SEA) bone marrow dendritic cells produce an exosome, an exosome is obtained by isolating and purifying and is used to treat a mouse with immune diseases, and then various indexes of this mouse such as body weight change, disease activity index, colon length, tissue lesions and pathological tissue score show that the disease of the experimental mouse is clearly better; it is noted that the exosome produced by japonicum egg antigen (SEA) bone marrow dendritic cells (BMDC) has clear therapeutic effect on immune diseases. The invention belongs to biological therapy, toxic and side effects of existing anti-inflammatory drugs are avoided maximally, the exosome has low toxicity and high efficiency, and a new method for treating immune diseases is provided.

Description

technical field [0001] The present invention relates to the field of medical and biological technologies, and more specifically relates to an exosome derived from dendritic cells stimulated by egg antigen of Schistosoma japonicum and its application. Background technique [0002] Inflammatory bowel disease is a group of chronic non-specific intestinal inflammatory diseases whose etiology is not yet clear, including ulcerative colitis (UC) and Crohn's disease (CD). Inflammatory bowel disease is a common disease in North America and Europe, and the incidence of inflammatory bowel disease in my country has gradually increased in the past 30 years. At present, the treatment of inflammatory bowel disease is mainly drug therapy, which mainly focuses on the application of hormones and immunosuppressants, but hormone therapy and immunosuppressants have certain side effects, and will increase the susceptibility of patients to infection and tumors. Therefore, it is an urgent need for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784A61K35/15A61P29/00A61P1/00
CPCA61K35/15C12N5/0639Y02A50/30
Inventor 孙希吴忠道王立富
Owner SUN YAT SEN UNIV
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