| Objective: Allergic reactions are mediated by immunoglobulin E(IgE)in serum and produce a variety of clinical manifestations,such as urticaria and angioneurotic edema,through the release of bioactive mediators by the effector cells.Sometimes,even the trace protein may cause fatal reactions for sensitive individuals.With more studies on allergic diseases,the cross-allergic reactions have attracted increasing attention.The cross allergy is due to an increased probability of the same epitope when two proteins have highly homological sequences or structures.To investigate the possibility and type of proteins of cross-allergic reaction between house dust mite(HDM)and egg white;the molecular components of HDM and egg white;and to compare the coincidence of the results by different immunoblotting methods,we were carried out 3 types of Considering to the high morbidity of HDM and egg white allergies and the difficulty in diagnosis and prevention,we did the results of the present study in order to providing a viable treatment,guiding proper diet and lifestyle for patients.Methods:1 Epidemiological investigation and analysis: an epidemiological investigation and a retrospective analysis were performed on these patients checked for allergies to specific allergens in the Dept.of Dermatology,Fourth Hospital of Hebei Medical University from Jan.2015 to Oct.2016.2 The serums of positive sIgE to HDM and egg white were selected for slot blot test,respectively.The result of slot blot test was compared with that of the immunoblotting test designed and saled by Acon Biotechnology Co.,Ltd.3 The cross-allergic reaction was detected by slot-blot test using the bloted HDM antigen and the serum of the positive sIgE to egg white.4 The pre-reaction,addding the egg whit antigens to the serum of the positive sIgE to HDM,was performed before the serum reacting with the bloted HDM antigens.The cross inhibition testwas carried out by slot-blot test in order to clarifying the cross-inhibition rate and effective inhibition concentration.5 Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE)was performed to detect the integrity,molecular weight and proportions of the proteins of HDM and egg white.6 Western blot(WB)was carried out to detect the antigenicity of HDM and egg white.Ten serums of positive sIgE to HDM and eggh white were selected to detect the antigenicity,respectively.Results: 1 Epidemiological survey 1.1 We retrospectively analyzed the serum sIgE results of patients diagnosed with allergic diseases in the Dept.of Dermatology,Fourth Hospital of Hebei Medical University from Jan.2015 to Oct.2016.Totally there were 5,186 patients registered,including 3,828 positive total Ig E(t IgE)patients.There are man 3100,woman 2086,with an average age 31.72±1.12.The course of disease ranged from 1 week to many years.The main diseases are urticaria,eczema and dermatitis.There are 1223 patients of positive sIgE to dust mite and 822 patients of positive sIgE to egg white.Among them,419 patients were simultaneously positive to dust mite and egg white,accounted for up to 34% of the patients of positive sIgE to dust mite.1.2 Distribution of the patients of positive sIgE: 1223(31.9%)for dust mite;822(21.5%)for egg white;430(11.2%)for milk;344(9%)for beef and mutton;223(5.8%)for wheat;300(7.8%)for cat and dog hairs;172(4.5%)for mold;166(4.3%)for dwarf ragweed;114(3%)for tree combinations;109(2.8%)for cockroaches;96(2.5%)for humulus;159(4.2%)for cashews and peanuts;152(4%)for fish and shrimp;and 150(3.9%)for mango.2 Comparative Analysis of the results of immunoblotting test(Acon Biotechnology Co.,Ltd.)and slot-blot testIn the 60 positive sIgE serums to dust mite,selected by immunoblotting test(Acon Biotechnology Co.,Ltd.),56 serums were detected positive by slot-blot test.The rate of consistency was 95%.3 The probability of the cross-allergic reaction between egg white and dust mite(slot-blot test)60 samples were randomly selected.There were 30 samples were observed positive,and the positive rate was 50%.4 The concentration-dependent inhibition in HDM by egg white was founded,and the maximum inhibition rate was 93% when the concentration of egg white was 100?g/ml.5 Analysis of protein components of dust mite and egg white 5.1 The three main bands of HDM were 57 kd,50kd and 16 kd,accounting for 55.5%,43.4% and 1.3% respectively.The 57 kd protein had the largest proportion.5.2 The four main bands of egg white were 75 kd,44.5kd,28 kd,and 14.3kd,accounting for 19.6%,66%,7.7%,and 6.6% respectively.The 44.5kd protein had the largest proportion.6 Antigenic analyses of HDM and egg white(WB)6.1 Two bands of HDM were observed at50 kd and 57kd;Derp10(myosin)and Derp4(?-amylase)were considered.6.2 Three bands ofegg white observed at 14.3kd,44.5kd and 75 kd,respectively;lysozyme(Lys),ovalbumin(OVA)and ovary transferrin(OVT)were considered.Conclusions:1 HDM and egg white are the main allergens of the allergic patients visited in our hospital;2 The main allergens of HDM are Derp10(protoplasmic protein)and Derp4(?-amylase).The main allergens of egg white are lysozyme(Lys),ovomucoid(OVM),ovalbumin(OVA),and ovary transferrin(OVT);3 There are similar and specific epitopes on the antigens of HDM and egg white;4 The cross-allergic reaction of HDM and egg white was probably due to the similar epitopes;5 The concentration-dependent inhibition in HDM by egg white was founded,and the maximum inhibition rate was 93% when the concentration of egg white was 100?g/ml. |
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