CD23 Surface Density On B Cells Determines Breg-facilitated IL-10 Secretion,as Well As Activation Of T Cells

Background and ObjectiveAllergic rhinitis(AR)is one of the most common nasal diseases caused by allergen uptake by antigen-presenting cells.IgE-mediated allergic rhinitis caused by the polarization of Th2 transmitted diseases,and was closely linked to allergic asthma.Recent years,the incidence of AR has risen significantly in most countries.According to the statistics,the average global incidence is 10%-40%,and 11%in China.The burden of disease is getting more and more serious,and becomimg a global health problem.IgE is known to have two distinct receptors,the high affinity receptor Fc?RI and the low affinity receptor CD23(Fc?RII).CD23 can expresse on the surface of B cells,monocytes,T cells,dendritic cells,platelets and other cells,mainly in the form of cell membrane surface and soluble forms.There is growing evidence that CD23plays an important role in the control of the activation of allergen-specific T cells by IgE-promoted antigen presentation.Recent studies have found that CD23 surface density on B cells determines the IgE-promoted allergic response.The original presentation led to the activation of allergen-specific T cells.Regulatory B cells(Bregs)are B cell populations that produce interleukin 10(IL-10)and exert a negative immunomodulatory effect.In the past two decades,researchers have discovered other immunological functions that Bregs possess,including antigen presentation,secretion of various cytokines,and production of IL-10 as an immunosuppressive agent.The negative immunoregulatory effects of Bregs were confirmed in animal models of experimental immunological meningitis,collagen-induced arthritis,chronic colitis,and multiple sclerosis.In allergic diseases,Bregs can promote the differentiation of regulatory T cells(Tregs)or directly suppress immune T cells by secreting cytokines such as IL-10 and transforming growth factor-?(TGF-?)proliferation.T helper cells are classified as type 1(Th1)and type 2(Th2),which are the major responses of the adaptive immune system.The secretion of cytokines,effector cells,and immune proteins are different.T helper cells mainly secrete IL-2 and IFN-?during Th1-predominant reaction;T-helper cells secrete mainly IL-4,IL-5 and IL-13 during Th2-dominated reactions.It can produce IgE and eosinophils,basophils as the main effector cells.Allergic rhinitis is Th2 reaction-based allergy.The aim of this study was to elucidate the role of IgE,IL-10,IL-4 and IFN-?in CD23 expression on B cells and their roles in the development and progression of allergic rhinitis associated with regulatory B cells and T cell activation.Animal and MethodsIn the first phase of animal experiment,48 SPF female SD rats were randomly divided into blank control group(group A,RA),allergic rhinitis group(group B,RB),anti-CD23 intervention group 1(group C1,RC1),Anti-CD23 intervention group 2(C2 group,RC2),12 rats in each group.On days 1,5,14 and 21,RA was treated with intraperitoneal injection of aluminum hydroxide adjuvant as a negative control,as well as RB and RC were treated with aluminum hydroxide and ovalbumin intraperitoneally;on 35 day,RA with saline nasal drops as a negative control,RB and RC were treated with 10%ovalbumin saline solution intranasal per day;RC in nasal ovalbumin saline solution nasal stimulation in the first 30 minutes,RC1 was given anti-CD23 antibody intranasal treatment per day;RC2 was given anti-CD23antibody in right leg intradermal injection per day.After the last stimulation,the nasal symptoms of rats in each group were observed and recorded within the prescribed time(30 min).After the completion of recording,RA was given physiological saline plus and 1%Evens blue mixture 0.1ml dorsal intradermal injection as a negative control,RB and RC was given 1mg ovalbumin plus and 1%Evans blue mixture 0.1ml intradermal injection,recorded local skin induration and dye diffusion mainwhile.After chloral hydrate anesthesia abdominal aorta blood,then rats were killed by cervical dislocation.The histological changes and eosinophilic infiltration of nasal mucosa were observed by HE staining.The expression of CD23,CD19 and IL-10 in nasal mucosa of four groups were detected by immunofluorescence.Serum total IgE,OVA-IgE,IL-10,IL-4 and IFN-?were detected by enzyme linked immunosorbent assay(ELISA).Flow cytometry and BD QuantiBRITE PE beads were used to detect the percentage of CD19~+CD23~+,CD19~+CD5~+CD1d~+B cells in the blood and the percentage of CD3~+CD4~+T cells,and the surface density of CD23 on the surface of B cells.In the second phase of clinical trials,we collected 40 inpatients with nasal diseases in our hospital.Among them,20 patients with isolated nasal septum deviation served as control group(Group A,HA)and 20 patients with allergic rhinitis as experimental group(Group B,HB).All were based on The annual diagnosis and treatment guideline for allergic rhinitis(Tianjin,2015)was diagnosed or excluded,and the nasal septum mucosa and venous blood were taken.HE staining was used to observe the histological changes of the nasal mucosa and eosinophil infiltration.Immunofluorescence was used to detect the expression of CD23,CD19and IL-10 in the nasal mucosa of the two groups.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of total IgE,IL-10,IL-4 and IFN-?in the serum of the two groups.The percentage of CD19~+CD23~+,CD19~+CD5~+CD1d~+B cells on the surface of B cells and the surface density of CD23 on B cells and the percentage of CD3~+CD4~+T cells in blood were measured by flow combined BD QuantiBRITE PE beads.ResultsCompared with RA,the number of sneezing and scratching nose in RB increased significantly;subcutaneous induration(±),dye diffusion;nasal mucosa HE staining showed a large number of eosinophils.The expression of CD23 and CD19,IL-10 were decreased by immunofluorescence.The ELISA showed that the total IgE,OVA-IgE and IL-4 were increased.The percentage of CD19~+CD5~+CD1d~+B cells was decreased and the percentage of CD19~+CD23~+B cells was increased.The density of CD23 distribution on the surface of B cells increased and the percentage of CD3~+CD4~+T cells was increased.Compared with the group of RB,the numbers of sneezing and nose-flexing in RC were decreased;subcutaneous induration(±),dye diffusion;eosinophilic and decreased hypertrophic Cell infiltration,the extent of arteriovenous dilatation and basement membrane destruction weakened;immunofluorescence CD23 expression was reduced,CD19,IL-10 expression slightly increased;ELISA test showed that the total IgE,OVA-IgE,IL-4,IL-10 and IFN-?slightly increased;the percentage of CD19~+CD5~+CD1d~+B cells increased,the percentage of CD19~+CD23~+B cells and CD3~+CD4~+T cells was decreased.Compared withRC2,RC1 had similar improvement in local symptoms,while blood markers did not decrease significantly in RC1,and serological markers in RC2improved significantly.Compared with HA,there were a large number of eosinophils and mast cells infiltration in nasal mucosa of patients with HB,arteriovenous dilatation and basement membrane destruction,and CD23 expression was found by immunofluorescence.The expression of CD19~+IL-10~+decreased with the increase of IgE,IgE,IL-4 and IL-10 and IFN-?.The percentage of CD19~+CD23~+B cells increased,the density of CD23 distribution on the surface of B cells increased,and the percentage of CD3~+CD4~+T cells increased.ConclusionThe surface density of CD23 on B cells correlates with IgE levels and may play an important role in the development of allergic rhinitis through IL-10-dependent T cell activation,which is dependent on Bregs secretion.The anti-CD23 antibody may regulate the symptoms and signs of allergic rhinitis,There was no significant difference between the application and nasal instillation in local symptoms.Instillation of nasal drops could be the preferred method for anti-CD23 antibody application.Anti-CD23 is expected to become a new method for the control and treatment of allergic rhinitis.