食物引起的过敏反应发病率正在增加,这对人们的健康构成了极大的威胁。为了评价牡蛎精氨酸激酶(arginine kinase,AK)过敏原的致敏性,利用AK口服灌胃和腹腔注射2种方式使BALB/c小鼠致敏。观察所有BALB/c小鼠的过敏反应、体重及血项指标情况,利用酶联免疫吸附法检测免疫球蛋白E(immunoglobulin E,IgE)的变化,使用苏木精-伊红及甲苯胺蓝染色观察脏器病变。结果表明,实验组小鼠都产生过敏反应,对照组无明显症状,实验组小鼠体重低于对照组,血液中细胞含量增加。此外,免疫期间实验组IgE含量都比对照组高,染色切片说明实验组小鼠组织出现较严重的损伤,空肠肥大细胞脱颗粒情况严重。该实验获得了牡蛎AK致敏血清,而且成功构建牡蛎AK的BALB/c小鼠过敏模型,为贝类过敏机制和临床免疫治疗提供了有意义的研究基础。
The incidence of food-induced allergic diseases is increasing, which poses a great threat to people's health. To evaluate the sensitivity of oyster allergen arginine kinase (AK), AK was intragastric and intraperitoneal injected to sensitize BALB/c mice to established mouse model. The allergic reaction, body weight, and blood index of all BALB/c mice were explored. The changes of IgE were detected by ELISA. The variations of tissues and organs were observed by hematoxylin-eosin (HE) and toluidine blue staining. The results revealed that all mice in the experimental group had allergic reaction, while the control group shows no obvious symptoms. The body weight of the experimental group was lower than that of the control group, and the number of cells in the blood increased. Furthermore, the IgE contents of the experimental group were higher than control group during immune period. The staining results showed that the experimental group displayed severe lesion damage in the organs, and the jejunal mast cells degranulation was serious. In this assay, AK sensitized serum is obtained, and the oyster AK BALB/c mouse allergy model is successfully constructed, which provides a significant research foundation for the shellfish allergy mechanism and clinical immunotherapy.
[1] SICHERER S H,SAMPSON H A.Food allergy:Epidemiology,pathogenesis,diagnosis,and treatment[J].Journal of Allergy and Clinical Immunology,2014,133(2):291-307.e5.
[2] WILD L G,LEHRER S B.Fish and shellfish allergy[J].Current Allergy and Asthma Reports,2005,5(1):74-79.
[3] ZHANG J T,LIU W Y,ZHANG R X,et al.Hypoallergenic mutants of the major oyster allergen Cra g 1 alleviate oyster tropomyosin allergenic potency[J].International Journal of Biological Macromolecules,2020,164:1 973-1 983.
[4] PEDROSA M,BOYANO-MARTNEZ T,GARC A-ARA C,et al.Shellfish allergy:A comprehensive review[J].Clinical Reviews in Allergy & Immunology,2015,49(2):203-216.
[5] LIANG Y L,CAO M J,SU W J,et al.Identification and characterisation of the major allergen of Chinese mitten crab(Eriocheir sinensis)[J].Food Chemistry,2008,111(4):998-1 003.
[6] GARCA-OROZCO K D,AISPURO-HERNNDEZ E,YEPIZ-PLASCENCIA G,et al.Molecular characterization of arginine kinase,an allergen from the shrimp Litopenaeus vannamei[J].International Archives of Allergy and Immunology,2007,144(1):23-28.
[7] AYUSO R,GRISHINA G,IB?EZ M D,et al.Sarcoplasmic calcium-binding protein is an EF-hand-type protein identified as a new shrimp allergen[J].Journal of Allergy and Clinical Immunology,2009,124(1):114-120.
[8] BEAVEN M A,METZGER H.Signal transduction by Fc receptors:The Fc&RI case[J].Immunology Today,1993,14(5):222-226.
[9] DVORAK A M.Mast cell-derived mediators of enhanced microvascular permeability,vascular permeability factor/vascular endothelial growth factor,histamine,and serotonin,cause leakage of macromolecules through a new endothelial cell permeability organelle,the vesiculo-vacuolar organelle[J].Chemical Immunology and Allergy,2005,85:185-204.
[10] RANC F,GRANDMOTTET X,GRANDJEAN H.Prevalence and main characteristics of schoolchildren diagnosed with food allergies in France[J].Clinical & Experimental Allergy,2005,35(2):167-172.
[11] GONZLEZ GALN I,GARCA MENAYA J M,JIMNEZ FERRERA G,et al.Anaphylactic shock to oysters and white fish with generalized urticaria to prawns and white fish[J].Allergologia et Immunopathologia,2002,30(5):300-303.
[12] CASTILLO R,DELGADO J,QUIRALTE J,et al.Food hypersensitivity among adult patients:Epidemiological and clinical aspects[J].Allergologia et.Immunopathologia,1996,24(3):93-97.
[13] LIU G M,LI B,YU H L,et al.Induction of mud crab(Scylla paramamosain) tropomyosin and arginine kinase specific hypersensitivity in BALB/c mice[J].Journal of the Science of Food and Agriculture,2012,92(2):232-238.
[14] 申洪.免疫组织化学染色定量方法研究(Ⅲ)[J].中国组织化学与细胞化学杂志,1995,4(1):89-92.
SHEN H.Study on the quantitative methods of immunohistochemistry(Ⅲ)[J].Chinese Journal of Histochemistry and Cytochemistry,1995,4(1):89-92.
[15] LEE J B,CHEN C Y,LIU B,et al.IL-25 and CD4+TH2 cells enhance type 2 innate lymphoid cell-derived IL-13 production,which promotes IgE-mediated experimental food allergy[J].Journal of Allergy and Clinical Immunology,2016,137(4):1 216-1 225.
[16] 郝海霞,李润花,殷国荣.霍乱毒素作为黏膜佐剂的研究进展[J].中国病原生物学杂志,2012,7(1):70-74;13.
HAO H X,LI R H,YIN G R.Advances in the study of cholera toxin as a mucosal adjuvant[J].Journal of Pathogen Biology,2012,7(1):70-74;13.
[17] LI X M,SCHOFIELD B H,HUANG C K,et al.A murine model of IgE-mediated cow’s milk hypersensitivity[J].Journal of Allergy and Clinical Immunology,1999,103(2):206-214.
[18] GONIPETA B,PARVATANENI S,TEMPELMAN R J,et al.An adjuvant-free mouse model to evaluate the allergenicity of milk whey protein[J].Journal of Dairy Science,2009,92(10):4 738-4 744.
[19] 田越.白细胞分类计数的临床意义分析[J].中国健康月刊,2011,30(5):168-169.
TIAN Y.Analysis of clinical significance of white blood cell classification[J].China Health Monthly,2011,30(5):168-169.
[20] ROTHENBERG M E,HOGAN S P.The eosinophil[J].Annual Review of Immunology,2006,24(1):147-174.
[21] FERNNDEZ-BLANCO J A,ESTVEZ J,SHEA-DONOHUE T,et al.Changes in epithelial barrier function in response to parasitic infection:Implications for IBD pathogenesis[J].Journal of Crohn’s and Colitis,2015,9(6):463-476.
[22] DHALIWAL B,PANG M O Y,YUAN D P,et al.Conformational plasticity at the IgE-binding site of the B-cell receptor CD23[J].Molecular Immunology,2013,56(4):693-697.
[23] BECKER J,OTT D,DIENER M.Impact of sensitization and inflammation on the interaction of mast cells with the intestinal epithelium in rats[J].Frontiers in Physiology,2019,10:329.