本文選題：腸道病毒71型　切入點：手足口病　出處：《重慶醫(yī)科大學》2012年碩士論文

【摘要】：背景：手足口�。╤and-foot-mouth disease, HFMD），是一種全球性常見的兒童傳染病，，多發(fā)生于學齡前兒童，癥狀主要為發(fā)熱和手、足、口腔以及臀部等部位的皰疹、丘疹、丘皰疹或者潰瘍，大多數(shù)患兒癥狀較輕微，部分患兒可能會有病毒性心肌炎、病毒性腦炎等并發(fā)癥，個別患兒可出現(xiàn)嚴重的并發(fā)癥而危及生命，如：腦干腦炎、無菌性腦膜炎、肺水腫、肺出血等。本病主要病原體為腸道病毒71型（EV71）和柯薩奇A組16型病毒（CoxA16）。而EV71所致患兒因其病程進展快、病情重、預后差，故引起了廣泛的關注。研究表明EV71感染后可能使患兒免疫功能受到抑制、發(fā)生紊亂。先天性免疫是機體抗病毒感染的第一道防線，NK細胞作為其關鍵細胞，在抗腸道病毒感染的過程中起著非常重要的作用，研究報道NK細胞功能低下或缺陷可發(fā)生嚴重的腸道病毒感染。 目的：檢測EV71所致手足口病患兒外周血中NK細胞比例、NK細胞亞群、激活型受體、抑制型受體及免疫效應分子陽性細胞率，研究EV71感染患兒NK細胞及功能的變化在EV71感染后機體免疫應答過程中可能的地位。 方法：試驗設2個實驗組，即：重癥病例組20例、普通病例組20例，1個對照組即：健康對照組30例。實驗組病人均為入院診斷為手足口病且經(jīng)RT-PCR檢測結果為EV71陽性的患兒；健康對照組為門診體檢兒童。年齡均為1～3歲。用流式細胞術檢測實驗組與對照組小兒外周血中NK細胞比例、NK細胞亞群（CD16+%NK、CD16+56+%NK、CD56+%NK）、NK細胞表面受體（NKp30、NKp46、NKG2D、CD94、NKG2A、CD107a）及NK細胞胞內免疫效應分子（PF、GrB和GNLY）陽性表達的細胞所占機體NK細胞的比率，采用單因素方差分析對結果進行統(tǒng)計學分析。 結果：重癥病例組外周血NK細胞比例較健康對照組明顯減少（P0.01），雖然普通病例組NK細胞數(shù)較重癥組高，但統(tǒng)計學分析無顯著差異（P0.05）；CD16~+%NK細胞亞群比例普通病例組均較健康對照組增高（P0.05），而重癥病例組較普通病例組低，但無統(tǒng)計學差異（P0.05）；重癥病例組、普通病例組激活型受體NKG2D陽性細胞百分率均較健康對照組低（P0.01），而抑制型受體CD94和NKG2A陽性細胞百分率均較健康對照組明顯增高（P0.01）；重癥病例組和普通病例組脫顆粒標記CD107a、PF、GrB、GNLY的陽性細胞百分率均較健康對照組低（P0.01或P0.05）。上述受體及免疫效應分子重癥病例組和普通病例組間無明顯差異（P0.05）；NKp30、NKp46陽性細胞率3組之間無明顯差異（P0.05）。 結論：EV71感染手足口病患兒外周血NK細胞比例及CD16~+NK細胞亞群減少；激活型受體NKG2D的表達下調、抑制型受體CD94和NKG2A的表達上調，說明NK細胞的活化受到抑制；脫顆粒標記受體CD107a表達降低及免疫效應分子（PF、GrB和GNLY）分泌降低，說明NK細胞的功能受到了抑制，這可能與EV71感染患兒臨床表現(xiàn)較重有關。重癥病例組的NK細胞比例較普通病例組有降低，此可能與重癥病例的發(fā)生有關。
[Abstract]:Background: hand, foot and mouth disease (HFMDD) is a common global infectious disease in children, mostly in preschool children. The symptoms are fever and herpes, papules, papules, herpes or ulcers in the hands, feet, mouth and buttocks, etc. Most of the children have mild symptoms, some of them may have complications such as viral myocarditis, viral encephalitis, and so on. Some children may have serious complications and endanger their lives, such as: brainstem encephalitis, aseptic meningitis, pulmonary edema, etc. The main pathogens of the disease were enterovirus 71 (EV71) and Coxsackie A group 16 virus (CoxA16C). However, the children caused by EV71 had a rapid progression, a severe illness and a poor prognosis. This has aroused widespread concern. Studies have shown that EV71 infection may inhibit the immune function of children and cause disorder. Congenital immunity is the first line of defense against virus infection, NK cells as its key cells, It plays a very important role in the process of anti-enterovirus infection. It has been reported that serious enterovirus infection can occur due to the low function or defect of NK cells. Objective: to detect the percentage of NK cell subsets, activated receptors, inhibitory receptors and immunoreactive molecule positive cells in peripheral blood of children with hand-foot-mouth disease induced by EV71. To study the possible role of NK cell and function in the immune response after EV71 infection in children with EV71 infection. Methods: two experimental groups were divided into two groups: severe cases group (n = 20), common case group (n = 20) and control group (n = 30). All the patients in the experimental group were diagnosed as hand-foot-mouth disease (HFMD) and were EV71 positive by RT-PCR test. The healthy control group was an outpatient check-up child. The age was 1 ~ 3 years old. Flow cytometry was used to detect the proportion of NK cells in peripheral blood of the experimental group and the control group. The NK cell subsets CD16 and NKP30, NKp46, NKG2D, CD94NKG2, CD107a, NK cell subsets, and NKP30, NKp4NKG2AtCD107a) and NK cell CD107a, respectively, were measured by flow cytometry. The percentage of NK cells positive for PFN GrB and GNLY in the body, Single factor analysis of variance (ANOVA) was used to analyze the results. Results: the percentage of NK cells in peripheral blood of the severe cases group was significantly lower than that of the healthy control group, although the number of NK cells in the common case group was higher than that in the severe case group. However, there was no significant difference in the proportion of NK cell subsets between P0.05and CD16- and the proportion of NK cell subsets in the common case group was higher than that in the healthy control group, while in the severe case group was lower than that in the normal case group, but there was no statistical difference between the two groups (P0.05). The percentage of activated receptor NKG2D positive cells in normal cases was lower than that in healthy controls, while the percentage of inhibitory receptor CD94 and NKG2A positive cells was significantly higher than that of healthy controls, and the degranulation of severe cases and normal cases was significantly higher than that of healthy controls. The percentage of positive cells of CD107a PFN GrBgnLY was lower than that of the healthy control group (P0.01 or P0.05). There was no significant difference in the percentage of NKp30NKp46 positive cells between the three groups. Conclusion the percentage of NK cells and CD16 ~ NK cell subsets in peripheral blood of children with HFMD infected with EV71 were decreased, the expression of activated receptor NKG2D was down-regulated, the expression of inhibitory receptor CD94 and NKG2A was up-regulated, which indicated that the activation of NK cells was inhibited. The decreased expression of degranulated receptor CD107a and the decreased secretion of immune-effector molecules, CD107a and GNLYB, suggested that the function of NK cells was inhibited. This may be related to the severe clinical manifestations of children with EV71 infection. The proportion of NK cells in severe cases is lower than that in normal cases, which may be related to the occurrence of severe cases.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R725.1

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