兒童時期是幽門桿菌感染發生率最高的時期，由學齡前、學齡兒童、到青少年時期是幽門桿菌抗體盛行率上升最快的時期。然而，有一部份兒童受到幽門桿菌感染後，會自動的把細菌從胃中清除(非易感者)；另外有些受感染者則形成持續性感染(易感者)。造成此現象的機制至今仍不清楚。我們研究室先前的動物研究發現，幼年感染幽門桿菌的小鼠，成年後約有一半小鼠會持續有幽門桿菌移生。持續有感染的小鼠，其胃上皮細胞缺乏IFN-γ的表現，但基質與單核球細胞則表現較強的IL-10。由此推測，宿主免疫細胞的調節功能，以及細胞激素產生的能力會與兒童幽門桿菌感染的易感性有密切的相關。因此，本研究的課題是要證實：兒童周邊血液CD4+CD25+與CD4+CD25– T細胞功能以及IFN-γ和IL-10的基因多型性與幽門桿菌的易感性有密切的相關性。
本研究族群共納入年齡介於4-12歲，40位感染以及40位未感染幽門桿菌，並提供新鮮血液樣本供調節性T細胞各項功能實驗的兒童；以及年齡介於4-12歲，79位感染以及127位未感染幽門桿菌之兒童，並提供DNA樣本供各項基因型分析。受試者FOXP3+調節性T細胞的比率使用流式細胞儀測得。評估T細胞功能為測定細胞激素的濃度。IL-10與IFN-γ單一基因多型性則以PCR、基因定序、以及Allele-specific PCR 測試。
首先觀察到感染幽門桿菌兒童周邊血液CD4+CD25high T細胞中表現Foxp3的比率顯著的高於未感染的控制組 (76.6% vs. 65.1%, P = 0.01)。同時在幽門桿菌感染兒童的胃antrum組織比未幽門桿菌感染之控制組有較多的FOXP3+細胞表現 (4.29 vs.1.31 units /400x HPF, P < 0.05)。感染幽門桿菌的兒童比起控制組不論在CD4+CD25– (286.2 vs.65.6 pg/mL, P < 0.05) 或CD4+CD25+ (5025 vs. 4350 pg/mL, P < 0.05) T細胞，分泌IFN-γ的能力皆比未感染的控制組顯著的高。控制組兒童周邊血液分離的CD4+CD25– T細胞被幽門桿菌抗原刺激後，IFN-γ產生的淨值顯著的高於有幽門桿菌感染兒童的細胞(104.8 vs. -85.5 pg/mL, P < 0.05)。相反的，在感染幽門桿菌兒童周邊血液分離出的CD4+CD25+ T 細胞，被幽門桿菌抗原刺激後，產生TGF-β1的淨值顯著的高於未感染的控制組(870.6 vs. -482.4 pg/mL, P < 0.05)。
本研究第二部份探討的是IFN-γ (+874 A/T＆microsatellite CA repeats)及IL-10 (-1082A/G, -819T/C, -592A/C)的基因多型性是否與兒童感染幽門桿菌有關。結果顯示在幽門桿菌感染及控制組兒童之間，IFN-γ與IL-10基因序列的單一基因多型性及IFN-γ microsatellite CA重複次數皆沒有顯著的相關性。
本研究是第一篇在人類小孩的血液樣本中觀察到調節性T細胞的功能與幽門桿菌感染有顯著關係的研究。綜合以上結果兒童幽門桿菌易感性與調節性T細胞有關而與IL-10 (-1082A/G, -819T/C, -592A/C)及IFN-γ (+874 A/T＆microsatellite CA repeats)基因多型性無關。

The highest infection rate of Helicobacter pylori (H. pylori) is reported in period of childhood. The increase rate of H. pylori antibody prevalence is the highest before teenage. After H.pylori infection, a group of children shows the auto clearance effect, and another group shows the persistent infection. So far, the mechanism is still not clear. In our previous animal study, we find persistent H. pylori infection in early age of mice is related to higher IL-10 functional expression but lower IFN-γ expression. We proposed that the regulatory functions in immunity and cytokine generation in host may be related to H. pylori infection in children. The aim of the study is focus on the relation among T cell functions of CD4+CD25+ and CD4+CD25– in child blood, genetic polymorphism of IFN-γ and IL-10, and H. pylori infection.
In the study, 79 infected and 127 uninfected subjects within 4 to 12 years old provided DNA samples for genetic analysis. In these subjects, 40 infected and 40 uninfected subjects provided blood samples for T cell related analysis. FOXP3+ Treg cell number was determined by flow cytometry. Cytokine secretion levels were used to evaluate T cell functions. The genetic polymorphism of IL-10 and IFN-γ were determined by using PCR.
FOXP3/CD4+CD25high expression in infected subjects was significantly higher than that in uninfected subjects (76.6 % vs. 65.1 %, P = 0.01). In antrum tissue, FOXP3+ number in infected subjects was significantly higher than that in uninfected subjects (4.29 vs. 1.31 units/400x HPF, P < 0.05). Both of CD4+CD25+ and CD4+CD25– cell, IFN-γ secreted level in infected subjects was significantly higher than that in uninfected subjects. Comparison of the cytokine net-change differences between before and after H. pylori antigen treatment was performed. In un-infected subjects, CD25- T cell with H. pylori protein treatment showed higher IFN-γ expression than those of H. pylori infected subjects (104.8 vs. -85.5 pg/mL, P < 0.05). In infected subjects, CD25+ T cell showed significantly higher TGF-β1 level than uninfected subjects (870.6 vs. -482.4 pg/mL, P < 0.05). In the part of IFN-γ and IL-10 genetic polymorphism, we did not find the relationship between genotypes and H. pylori infection.
It is the first study to observe the association between H. pylori infection and Treg cell functions in blood of human children. Accorrding the results, susceptibility of childhood H. pylori infection should be related to Treg function, but not related to genetic polymorphisms of IL-10 and IFN-γ.

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