Mechanism Of Memory B Cells Changes In Chronic Granulomatous Disease And Clinical Manifestations And Molecular Characteristics Of Autosomal Dominant Hyperige Syndrome

PART I STUDY ON THE REGULATION OF MEMORY B CELLS BY TOLLICULAR HELPER T CELLS IN CHRONIC GRANULOMATOUS DISEASEObjective:Chronic granulomatous disease（CGD）is a primary immunodeficiency disease with phagocytic dysfunction.In this study,we investigated the changes of number of memory B cell（MBC）and the possible mechanism in CGD through patients and gp91^phoxKO mice.Methods:We collected the clinical data of 15 patients with CGD,detected the percentage of B cell subsets,Tfh cells,Tfh cell subsets and the expression of PD-1,ICOS,CD40L on the surface of Tfh cells in CGD patients and healthy controls by flow cytometry.Next,we checked the function of gp91^phoxKO mice.Then,we immunized mice with TD-Ag NP-KLH.The percentage of B cell subsets,Tfh cells and its related functional molecules ICOS,PD-1 in the spleen of gp91^phoxKO and WT mice were detected by flow cytometry.Results:Compared with the healthy control group,the percentage of Na?ve B cells in CGD patients was significantly higher,the percentage of MBC,class switched MBC and PC in CGD patients were significantly lower.The percentage of Tfh in CGD patients was significantly lower than that in the healthy control group,and the percentage of Tfh17 cells was significantly higher than that in the control group.CD40L on the surface of Tfh cells in CGD patients was significantly lower than that in the control group,and there was no significant difference in the ratio of ICOS and PD-1 on the surface of Tfh cells.CD40 on the surface of B cells was not significantly different from that in the healthy control group.The respiratory burst dysfunction of gp91^phoxKO mice was consistent with that of CGD patients.There was no significant difference between WT mice and gp91^phoxKO mice in the percentage of B cell subpopulation and Tfh cells in the spleen.The spleen of gp91^phoxKO mice increased after primary and secondary immunization with T-cell-dependent antigen,but the percent of GCB cells,antigen-specific GCB cells,switched MBC,antigen-specific switched MBC,PC and Tfh cells showed no significant difference compared with WT mice.Conclusion:In CGD patients,the differentiation of Na?ve B cells into MBC and PC was blocked.The decreased number of Tfh cells and the expression of CD40L on the surface of Tfh cells may be related to the decrease of MBC.The respiratory burst dysfunction of gp91^phoxKO mice is consistent with that of CGD patients,but the response to TD-Ag is normal,which was inconsistent with the decrease of MBC in patients,suggesting that the response mechanism of NP-KLH in gp91^phoxKO mice was inconsistent with that in patients.PART II CLINICAL MANIFESTATIONS AND MOLECULAR CHARACTERISTICS OF AUTOSOMAL DOMINANT HYPERIGE SYNDROMEObjective: Autosomal dominant hyper IgE syndrome（AD-HIES）caused by STAT3 gene mutation is a rare primary immunodeficiency disease.To better understand the disease,we described the clinical and molecular characteristics,and Th17 cells of 20 AD-HIES patients in our center.Methods: We collected the clinical and laboratory examination data of 20 AD-HIES patients,sequenced the STAT3 gene of patients and parents,detected Th17 cells by flow cytometry.Results: Among 20 cases,14 were male,6 were female.The mean age at onset was 0.12 years.The mean age at diagnosis was 5.31 years.All patients had NIH scores over 40 points.The most common presentation was eczema（100%,20/20）,pneumonia（95%,19/20）,skin abscesses（85%,17/20）and chronic mucocutaneous candidiasis（70%,14/20）.Staphylococcus aureus（S.aureus,40%,6 / 15）is the main pathogen in pneumonia.Recurrent pneumonia was associated with secondary pneumatocele（50%,10/20）.Seven patients（38.8%,7/18）suffered complication from BCG vaccination.20 patients（100%,20/20）had high serum IgE level（757.4-38900 IU/ml）and eosinophil number（510-4180 cells/μL）.The numbers of Th l7 cells were significantly decreased in peripheral blood of patients with AD-HIES.Eleven STAT3 heterozygous mutations were identified,2 of which were not reported（C712G;c.1139+5G>T）.Three patients（15%,3/20）have died.Nine patients（45%,9/20）were known to be on antibiotic and antifungal prophylaxis,and 4 patients（20%,4/20）on Ig G treatment.Conclusion: Children with repeated pneumonia,especially S.aureus pneumonia,cold skin abscess,fungal infection,eczema,increased serum eosinophil and IgE level,should be alert to AD-HIES.In this study,our patients had earlier onset and diagnostic age,higher rate of BCG complications and higher incidence of mortality.Lifelong prophylactic sulfamethoxazole and itraconazole reduced infections.