Analysis Of Current Status Of IVIG Replacement Therapy In 114 Cases Of Congenital Agammaglobulinemia And Immunological Characteristics Of X-Linked Moesin-Associated Immunodeficiency

PART Ⅰ ANALYSIS OF CURRENT STATUS OF IVIG REPLACEMENT THERAPY IN 114 CASES OF CONGENITAL AGAMMAGLOBULINEMIAObjects: To explore the current status of intravenous immunoglobulin replacement therapy for congenital agammaglobulinemia,and provide a basis for reasonable and standardized treatment.Methods: The basic Information,clinical manifestations,laboratory examinations,treatment outcomes of 114 patients with congenital agammaglobulinemia in Children’s Hospital of Chongqing Medical University from 1988 to 2020 were retrospectively analyzed.To compare the improvement of clinical manifestations between regular and irregular infusions,χ2 test was used for data analysis.Patients were divided into non-cough,short-term cough and long-term cough group,chronic lung disease and non-chronic lung disease group and arthritis and non-arthritis group to analyze age at onset,age at diagnosis,delayed diagnosis time,initial time of immunoglobulin replacement,dose of immunoglobulin replacement,Ig G trough level and other indicators.T test,F test or non-parametric test were used for the comparison.After univariate analysis,two-category Logistic regression analysis was performed on the statistically significant variables.Results: All the patients were male(mean of onset age(22.3±17.9)months,mean of diagnosis age(89.3±53.7)months,diagnosis delay(63.3±46.1)months,initial time of immunoglobulin replacement(75.0±45.4)months).66 patients had been followed up to April 2020.Among 66 patients,42(63.6%)received regular infusion,whose monthly immunoglobulin replacement dose was(538.0±104.6)mg/kg per month and Ig G trough level was(5.8±1.5)g/L.Whereas 24 patients(36.4%)were treated irregularly.There was no significant statistical difference in the decreased incidence rate in clinical manifestations of fever,cough,sinusitis,diarrhea,otitis media and arthritis between regular and irregular infusion.61(92.4%)of 66 patients had fever before infusion.The number of fever was significantly decreased after infusion [5.0(2.0,12.0)vs.0(0,1.0)per year(Z=-6.436,P<0.0001)].There were 60 patients(90.9%)suffered wet cough before treatment and 36(54.5%)after.Initial time of immunoglobulin replacement was significantly different between long-term cough(27 cases),short-term cough(9 cases)and non-cough group(18 cases)[96.6±51.4 vs.64.0±40.8 vs.63.4±42.0 months(F=3.554,P=0.035)].29patients(43.9%)were diagnosed with chronic lung disease.Initial time of immunoglobulin replacement,age at diagnosis,and delayed diagnosis time of chronic lung disease(29 cases)were significantly later and longer than those in children with non-chronic lung disease(34 cases)[107.0±48.1 vs.46.5(26.3,64.3)(Z=-4.33,P<0.0001)],[103.0(75,142.5)vs.47.0(31.0,68.0)months(Z=-3.486 P<0.0001)],[91.0(55.1,128.7)vs.29.5(10.3,41.5)months(Z=-4.386,P<0.0001)].In addition,among 114 patients,42patients(36.8%)were confirmed arthritis,32 patients followed up to April2020.Initial time of immunoglobulin replacement and delayed diagnosis time of arthritis(32 cases)were significantly later and longer than those in children with non-arthritis(34 cases).Conclusion: After immunoglobulin replacement therapy,the clinical symptoms such as fever,sinusitis,diarrhea,and otitis media have been improved to varying degrees.However,long-term wet cough,chronic lung disease,and arthritis were still prominent clinical problems.Standard immunoglobulin replacement therapy had enabled some patients’ disease be effectively controlled.PART Ⅱ IMMUNOLOGICAL CHARACTERISTICS OF X-LINKED MOESIN-ASSOCIATED IMMUNODEFICIENCYObjects: We analyzed the clinical and immunological Characteristics of a Chinese boy with MSN mutation,to extending the previously described phenotypes.Methods: Collect the basic information,clinical data and peripheral blood samples of a child with X-MAID in the Children’s Hospital of Chongqing Medical University.Western blot was used to detect the Moesin protein expression of neutrophils and mononuclear cells,and flow cytometry was used to detect Moesin protein expression in various types of peripheral blood cells,lymphocyte apoptosis,F-actin aggregation function of neutrophils.Luminol chemiluminescence method was used to detect the function of neutrophils to produce reactive oxygen clusters.Results: The patient enrolled in this study was a 2y9 m male child,mainly manifested as repeated upper respiratory tract viral infections,refractory eczema and recurrent alopecia areata,persistent lymphocytopenia,repeated neutropenia,and PID series gene results showed MSN mutation(c.511C>T p.Arg171Trp).The mutation was derived from the mother.The proliferation of CD4+T,CD8+T and CD19+ cells was reduced.The TRECs were 285(291.6-429.5)cells per reaction.Western blotting indicated that the expression of Moesin protein in the PBMCs was significantly reduced,whereas that in neutrophils was normal.Flow cytometry showed that the Moesin expression of neutrophils,monocytes,and B cells was normal,that of CD4+ T cells decreased,and CD8+ T and NK cells did not express.As age increased,Moesin expression of T cells gradually decreased.Memory cells expressed less than native cells in CD4+T and CD19+B cells.Immunological phenotype analysis showed T,B,and NK cells were reduced,and the proportion of TERMA CD8+T increased significantly.Bone marrow appearance showed there was no abnormality in lymphocyte differentiation and maturation.However,CD4+T and CD8+T cells were found significantly increased apoptosis after 72 hours of CD3/CD28 stimulation.And after 16 hours of FAS stimulation,apoptosis of neutrophils was found to increase significantly.Neutrophil apoptosis in children was still increased with LPS stimulation.The production of reactive oxygen species by neutrophils in children was stronger than that of control.After F-actin depolymerization,the production of ROS in children was lower than normal.The polymerization of F-actin in neutrophils was not affected by the expression of Moesin protein.Conclusion: One child with X-MAID was diagnosed through clinical manifestations,immune function screening,and protein function testing,making it the eleventh case in the world.And this case expanded the immunological phenotype of the disease: NK cells do not express Moesin protein;The decrease of lymphocytes and neutrophils in children with X-MAID is related to the increase of apoptosis;The function of neutrophil F-actin polymerization is not affected;Under the mutation of MSN,there may be other pathways involved in neutrophil ROS shutdown.