| Section I The changes of CD4+CD25+Foxp+Treg cells andTh1/Th2cells in house dust mite subcutaneous specificimmunotherapyObjectiveThis study was to investigate the changes of CD4+CD25+Foxp3+regulatory T (nTreg)cells, Th1/Th2cells, and cytokines (IL-5, IL-10, IL-17, IFN-γ and TNF-α) in the housedust mite subcutaneous specific immunotherapy (HDM-SCIT), and to discuss the possiblemechanisms of HDM-SCIT.MethodsWe enrolled27patients (male:16, female:11;25.7±11.8years of age) with mild tomoderate asthma and/or rhinitis who had received standardized HDM-SCIT for one year(between December2011and December2013) in The First Affiliated Hospital,Guangzhou Medical University (SIT group),21patients (male:13, female:8;32.2±13.2years of age) with uncontrolled asthma and/or rhinitis (uncontrolled group),22patients(male:13, female9;28.0±9.5years of age) with controlled asthma and/or rhinitis(controlled group) and20healthy subjects (male:10, female:10;26.3±4.0years of age)(normal group).All of the subjects underwent skin prick test (SPT) against common airoallergens,pulmonary function tests for forced expiratory volume in1second (FEV1) and airwayresponsiveness to histamine bronchial provocation (AHR), peripheral eosinophil count,and detection of serum level for total IgE (TIgE), dermatophagoides (D.) pteronyssinusand D. farinae specific IgE (sIgE). Peripheral blood mononuclear cells (PBMCs) were isolated from all of the subjects. They were stimulated with HDM extracts for72hs. nTregcells were measured by flow cytometry. Levels of IL-5, IL-10, IL-17, IFN-γ and TNF-α insupernatants from allergen-stimulated cultures were measured by suspension chip.Patients with uncontrolled and controlled asthma were requested to finish asthma qualityof life questionnaires (AQLQs).Results1. FEV1%predicted value in the uncontrolled and controlled group was significantlylower than the normal group (p<0.01) while it did not have significant difference betweenthe SIT and normal group (p>0.05). AHR was found in all patient groups, but,uncontrolled and the controlled groups showed significantly higher grade of the AHR thanthe SIT group (p<0.001).2. There was no statistically significant difference among the four groups (p=0.219)with regards to the percentage of nTreg in PBMCs although it was high in SIT and normalgroup. Similarly, after vitro HDM-stimulation, the percentage of nTreg also did not havestatistical difference (p=0.392) among the four groups. The percentages of nTreg in4groups increased significantly after allergen stimulation with p<0.001for SIT group,p=0.007for uncontrolled group, p=0.007for controlled group and p=0.014for normalgroup. However, the magnitude of increase in4groups did not have significant difference(p=0.655).3. The percentages of Th1cells in the PBMCs among4groups had no significantdifference before (p=0.541) and after (p=0.906) allergen stimulation. In comparison withthe baseline value, the percentages of Th1cells in4groups had no significant changes(p=0.448for SIT group, p=0.942for uncontrolled group, p=0.590for controlled group andp=0.125for normal group) after allergen stimulation.4. The percentage of Th2cells in PBMCs in SIT group and uncontrolled group wassignificantly higher than in normal group (p=0.011and p=0.028, respectively) while there was no significant difference between other groups. After allergen stimulation, thepercentages of Th2cells in uncontrolled group was significantly higher than in normalgroup (p=0.03), while there was no significant difference between other groups. Incomparison with the baseline value, only SIT group showed significant decrease for thepercentage of Th2cells after allergen stimulation (p=0.008) while no such change wasfound in other groups.5. The ratio of Th1/Th2in PBMCs in healthy group was significantly higher than SITgroup (p=0.004), uncontrolled group (p=0.044) and controlled group (p=0.029). It had nosignificant difference among4groups after allergen stimulation. In comparison with thebaseline value, the ratio of Th1/Th2in SIT and controlled group increased significantly(p=0.03and p=0.045, respectively) while the other two groups had no significant change.6. Before HDM stimulation, the level of IL-5and IL-10in uncontrolled group wassignificantly higher than in SIT group (p=0.032, p=0.044), controlled group (p=0.01,p=0.003) and normal group (p=0.024, p=0.037), whereas there was no significantdifference for level of IL-17, IFN-γ and TNF-α among all4groups. After allergenstimulation, the level of IL-5, IL-17and TNF-α in normal group were significantly lowerthan SIT group (p<0.05), uncontrolled group (p<0.05) and controlled group (p<0.05),while no significant difference was found for the level of IFN-γ among4groups. Incomparison with the value before stimulation, the levels of all five cytokines in SIT groupwere increased significantly (p<0.01). In normal group, the level of IFN-γ and TNF-αwere increased significantly (p<0.01), while the level of IL-5, IL-10and IL-17had nosignificant changes. In uncontrolled group, the level of IL-5, IL-10and IL-17increasedsignificantly (p<0.01), the level of IFN-γ and TNF-α had no significant changes. And incontrolled group, the levels of all five kinds of cytokines were increased significantly(p<0.01) except for IFN-γ.7. Level of IL-17in culture supernatant was significantly increased in patients withAHR in SIT group than those without AHR after allergen stimulation (p=0.035). It was positively related with the peripheral EOS count in（r=0.559, p=0.03）and TIgE (r=0.685,p=0.005) with linear regression analysis. The percentage of Th1cells correlated positivelywith percentage of nTreg cells before (r=0.597, p=0.013) and after (r=0.485, p=0.048)allergen stimulation.8. FEV1%predicted was positively related to the level of culture supernatant IFN-γ(r=0.483, p=0.019), the magnitude of increase of the level of culture supernatant IL-10(r=0.378, p=0.047) and ratio of Th1/Th2(r=0.523, p=0.013) after allergen stimulationafter allergen stimulation in patients with routine medication treatment. The scores ofAQLQs was positively related to the magnitude of increase of percentage nTreg cells afterallergen stimulation (r=0.539, p=0.007).Conclusion1. One year standardized HDM-SCIT can improve pulmonary function includingFEV1%predicted value and AHR, rebalance Th1and Th2in patients with allergic asthmaand/or rhinitis.2. The clinical efficacy of HDM-SCIT was associated with the improvement ofTh1/Th2ratio. Monitoring the Th1/Th2ratio in PBMCs and its response to HDM may beuseful for predicting clinical efficacy of HDM-SCIT.3. The levels of IL-5, IL-10and IL-17in asthma patients increased significantly,while the levels of IFN-γ and TNF-α increased in the healthy subjects upon allergenstimulation.4. One year HDM-SCIT could up-regulate nTreg cells that may play some role inre-balance Th2and Th1, but, further studies are needed. Section II Immediate Adverse Reactions duringSubcutaneous Standardized House Dust Mite SpecificImmunotherapy in Patients with Asthma and/or RhinitisObjectiveTo analyze immediate adverse reactions during subcutaneous specificimmunotherapy (SCIT) in mite allergic patients with asthma and/or rhinitis bystandardized house dust mite allergen vaccine.MethodsRetrospective evaluation of immediate adverse reactions in462patients whoreceived subcutaneous standardized house dust mite specific immunotherapy betweenNovember2004and May2012was performed. The relationship between immediateadverse reactions and SCIT dosage, age, gender, diagnosis, and levels of serum IgE wasanalyzed.Result1. Of462patients who received SCIT,452(97.8%) had immediate local reactions,153(33.1%) had immediate systemic reactions. Among15645injections,8523(54.5%)developed immediate local adverse reactions, and397(2.5%) had immediate systemicadverse reactions.2. Frequency of immediate local adverse reaction was increased with increase ofSCIT dosage. The immediate systemic adverse reactions were mainly occurred in theup-dosing phase between2000and80000SQU.3. In comparisons between different diagnosis, the prevalence of immediate adversereactions in patients with only asthma and asthma with rhinitis was significantly higherthan that in patients with only allergic rhinitis (p<0.01).4. The prevalence of immediate local adverse reaction in children (≤14years) wassignificantly higher than that in adults (p<0.001). 5. The prevalence of immediate adverse reactions in boys was significantly higherthan in girls (p<0.01), while it was significantly higher in female than male in adults(p<0.01).6. Prevalence of immediate adverse reactions (local and systemic) increasedsignificantly with the increase of serum Dermatophagoides pteronyssinus IgE level(p<0.001). The patients who developed immediate systemic adverse reactions hadsignificantly increased total IgE level than those who did not (p=0.048).ConclusionImmediate local adverse reactions are common during subcutaneous specificimmunotherapy with standardized house dust mite vaccine. However, immediate systemicadverse reactions are rare. Patients with asthma, adult female, juvenile male and highlevels of house dust mite specific IgE and total IgE are risk factors for the development ofimmediate adverse reaction. Much attention should be paid for monitoring severe systemicreactions during up-dosing phase.General conclusion of the study1. SCIT is effective and safety for house dust mite allergic asthma and (or) rhinitispatients, but much attention should be paid for monitoring severe systemic reactionsduring up-dosing phase.2. The clinical efficacy of HDM-SCIT was associated with the improvement ofTh1/Th2ratio. Monitoring the Th1/Th2ratio in PBMCs and its response to HDM may beuseful for predicting clinical efficacy of HDM-SCIT.3. The levels of IL-5, IL-10and IL-17in asthma patients increased significantly,while the levels of IFN-γ and TNF-α increased in the healthy subjects upon allergenstimulation. 4. One year HDM-SCIT could up-regulate nTreg cells that may play some role inre-balance Th2and Th1, but, further studies are needed. |
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