The Role Of B Cells In Allergic Diseases And Autoimmune Diseases Based On Multiple Analyses Of Antibodies And The Correlation With HLA Alleles

Background: Allergic diseases and autoimmune diseases are multifactorial disorders with genetic,immune,environmental and other factors implicated in aetiopathogenesis.Incidence of them is increasing in recent years.They are both very serious public health problems.Researchers focus on the etiology and pathogenesis of the two diseases,especially from the perspective of immunity and heredity,which is very significant for the diagnosis and treatment of the diseases.The pathological antibodies including sIgE and multiple autoantibodies are related to occurrence of allergic diseases and autoimmune diseases.These pathological antibodies derive from B cells and plasma cells.Union of pathologic antibody and the antigen(allergen and autoantigen)will lead to a series of pathological reactions(cytokines releasing,cell activation,activation of complement system,etc.),eventually leading to organ damage.Pathological antibodies play an important role in the diagnosis of the disease in some autoimmune diseases.Many autoantibodies can be detectable several years prior to clinical symptoms or definite diagnosis.Therefore,detection of autoantibodies can be used as the risk assessment of disease in asymptomatic patients.In clinical practice,both conditions that a single pathological antibody is positive or a various pathological antibodies are positive can be found in patients.It is unclear that different pathological antibodies are synthesized randomly or necessarily.Among these different types of pathological antibodies,concentrations are diverse.Diagnostic and therapic value of pathological antibodies with highest concentration was rarely studied.In recent years,generation of pathological antibodies was studied widely.Many cells including memory B cells,plasma cells,autoreactive B cells and the class switchof immunoglobulin were research focus.In allergic diseases and autoimmune diseases,stimulation of antigen(allergen and autoantigen)is closely related to the synthesis of pathological antibody.Meanwhile,antigen processing and antigen presentation are closely related to HLA.In recent decades,studies have identified many HLA gene polymorphisms for susceptibility of allergic diseases and autoimmune diseases.In allergic diseases,HLA-DRA was found associated with some drug induced allergy,such as penicillin.For various types of inhalation and food allergen,it was demonstrated that HLA-? allele was associated with house dust,cockroaches,food additives,strawberries,trees in childhood asthma.With the development of technology used in pathological antibodies detection,a variety of HLA alleles associated with different allergens have been found,including HLA DQA1 * 01:01,HLA DQA1 *06:01,HLA DQB1 * 03:03,HLA DQB1 * 06:01 is closely related to allergy of the mites,HLA-DRB1 * 01,HLA DQB1 * 05:01 was associated with allergy of mugwort.Meanwhile,some HLA alleles have not been found for some allergens.In SLE,it was reported that the HLA-? allele especially HLA DQA1,DRB1,DQB1 plays important roles.HLA-DRB1*15:01,HLA-DQA1*01:02,HLA-DQB1*06:02,DRB1*03:01,DQA1*05:01,DQB1*02:01,HLA-A*03:01,HLA-B*07:02,HLA-C*07:02,HLA-A*01:01,HLA-B*08:01,HLA-C*07:01 are all related genetic segment for SLE susceptibility.Genetic factors play an important role in the pathogenesis of these two diseases.Many other factors were also involved.It is rarely reported on the correlation between HLA alleles and variety of pathologic antibody.Objective: This paper aims to compare observed proportion of mono-pathological antibody and theoretical proportion of mono-pathological antibody through the analysis of pathological antibody data in allergic diseases and autoimmune disease and evaluate diagnostic value of trait antibody in diseases.Various types of unrelated antibodies in the serum were detected.Another objective of this paper is to analyse the relation between the relative length of lymphocyte telomeres and the varieties of pathological antibody,and to study the relation between HLA-DRB1,HLA-DQB1 allele and varieties of pathological antibody.Methods: 1.Serum inhalation and food allergen sIgE antibody(linear western blot method,LIA)in 2409 patients with allergic diseases were detected.Theoretical proportions of mono-sensitization(TPM)were put forward and calculated according to principle of probability theory.The observed proportions of mono-sensitization(OPM)7w ere compared with TPM.Trait sIgE was defined based on intensity.Diagnostic efficiency was evaluated by ROC analysis.2.Serum ANAIgG were detected in 643 patients with autoimmune diseases(LIA).Theoretical proportions of monospecific ANA(TPM)were put forward and calculated.The observed proportions of mono-specific ANAs(OPM)were compared with TPM.Trait ANAIgG was defined based on intensity.Diagnostic efficiency of trait ANAIgG was evaluated by ROC analysis.3.Infectious antibodies including IgM antibody of Toxoplasma gondii(TOXO)cytomegalovirus(CMV),rubella virus(RV),IgG andtibody of hepatitis C virus(HCV),Treponema pallidum(TP),human immunodeficiency virus(HIV),together with ANAIgG and sIgE were detected by using chemiluminescent microparticle immunoassay,electrochemiluminescence assay,ELISA and LIA method in 55 patients with allergic diseases and 171 patients with autoimmune disease.4.Lymphocytes telomere relative length was detected in 33 patients with allergic diseases and 36 cases of SLE patients together with healthy control group matched by age and sex by using fluorescent quantitative PCR.5.HLA-DRB1 and HLA DQB1 alleles were detected in100 cases of allergic diseases and 100 patients with SLE by using sequence-based typing method(PCR-SBT).Based on searching online database and Discovery Studio(DS)software,correlation between variety of pathological antibodies and HLA alleles was analyzed.Another correlation between between incidence and HLA alleles was also analyzed.Sequence alignments were made for amino acid of related HLA alleles and unrelated HLA.Molecular models were built for HLA?chain.Stability of related HLA?chain was compared with unrelated HLA?chain.Results: 1.OPM and TPM were compared in multiple perspectives(gender,age,season,type of disease and allergy pathway),indicating that OPM was significantly lower than TPM(p<0.001)in the allergic diseases.The area under ROC curve(AUC)of trait sIgE was 0.749.2.OPM was significantly lower than TPM(p<0.001)in autoimmune disease group.AUC of SLE(systemic lupus erythematosus)was 0.916.AUC of the rheumatoid arthritis group was 0.646.AUC of other autoimmune diseases group was 0.830.3.There was a statistical significance in the difference of anti CMV IgM antibody and the anti TP IgG antibody in allergic diseases compared with healthy control group(p<0.001).There was a statistical significance in the difference of anti TOXO IgM antibody,anti TP IgG antibody and anti HCV IgG antibody in autoimmune diseases compared with healthy control group(p< 0.05).In autoimmune diseases the8 average variety of IgE(0.88)was higher than control group(0.20)(p< 0.001).In allergic diseases group the average variety of IgE(2.37)was higher than control group(p<0.001).In autoimmune disease group the average variety of ANAIgG(2.47)was higher than control group(0.05)(p<0.001).In allergic diseases the average variety of ANAIgG(0.25)was higher than control group(p=0.04).4.There were statistical significances in the differences of lymphocyte telomeres relative length in SLE compared with healthy control group(p=0.011).There was a statistical significance in the differences of lymphocyte telomeres relative length in allergic diseases compared with healthy control group(p=0.043).The telomeres relative lengths of two types of diseases were less than healthy control group.Telomeres relative length showed negative correlation with variety of ANAIgG in SLE(B=-0.249,p=0.044),it also showed negative correlation with the variety of s IgE in allergic diseases(B=-0.250,p=0.048).5.Linear regression analysis was made for high-resolution HLA alleles and the variety of pathological antibodies.Logistic regression analysis was made for high-resolution HLA alleles and the incidence of allergic diseases and SLE.Results showed that the variety of pathological antibodies was positively related to HLA-DRB1* 01:02 and the incidence was positively related to HLA-DRB1 *07:01 in allergic diseases.It was demonstrated that the variety of pathological antibodies were positively related to HLA-DQB1*02:02,HLA-DQB1*04:01,HLA-DRB1*07:01,HLA-DRB1*08:03,HLA-DRB1*12:10 and the incidence was related to HLA-DRB1*08:03,HLA-DRB1*15:01 in SLE.Tertiary structures of Beta chain were built based on the comparison of sequence alignment between relevant and irrelevant HLADRB1 and HLADQB1.The systemic energy was calculated by using Minimization.Evaluation of stability of molecules showed that the stability of HLADRB1 and HLADQB1 molecules associated with variety of pathological antibodies or incidences of disease was lower than that of unrelated HLA molecules.Conclusion: 1.High proportion of multiple pathological antibodies is inevitable rather than random in allergic diseases and autoimmune diseases.The abnormal activation and proliferation of B cells is the predominant factor in the generation of multiple pathologic antibodies.Trait pathological antibodies(sIgE/ANAIgG)have good diagnostic efficacy for allergic diseases and SLE.2.Some antibodies were higher in allergic disease than that of healthy control group.In autoimmune diseases some antibodies were higher than that of healthy control group.It suggested that in two typesof diseases,both pathological antibodies that are specific in certain diseases and some unspecific antibodies are abnormal.Therefore,disorders of B cell function may be the basis of allergic disease and autoimmune disease.Extensive cloning of B cells is common.Specificity of activation and proliferation may be poor.It results in pathological damage in multiple organs.3.In the detection of the relative length of telomeres for patients with multiple pathological antibodies,results were lower in allergic diseases and SLE than that of healthy control group.They were negatively correlated with the variety of pathological antibodies.It is suggested that the synthesis of polyclonal antibodies is the result of excessive activation and proliferation of B cell.It affects aging of B cell.4.In this study,some HLA-DRB1 and HLA-DQB1 are associated with diseases and the variety of pathological antibodies.The stability of beta chain of HLA-DRB1 and HLA-DQB1 molecules is lower than that of unrelated HLA molecules.Most differences sites of these two kinds of HLA molecules distributed in beta chain antigen binding site.Thus for related HLA molecules and unrelated HLA molecules,binding specificity of antigen peptide and affinity may be different based on molecular structure.The antibodies produced by B cell may be affected,which leads to the changes of incidence of diseases and the variety of pathological antibodies.