重组幽门螺杆菌疫苗诱导小鼠产生THL型保护性应答及免疫后胃炎

目的 研究以减毒鼠伤寒沙门氏菌为载体构建的重组幽门螺杆素酶A亚单位及过氧化氢酶、／尿素酶A亚单位融合蛋白疫苗对小鼠的免疫保护作用及相关机制。方法 将表达幽门螺杆菌(Helicobacter pylori,HP)尿素酶A亚单位(UreA)、过氧化氢酶(KatA)及尿素酶A亚单位／过氧化氢酶融合蛋白(Urea／KatA)的减毒鼠伤寒沙门氏菌(Salmonella typhimurium)分别灌胃免疫小鼠,另设单纯减毒鼠伤寒沙门氏菌免疫鼠和生理盐水免疫鼠为对照。免疫4周后以幽门螺杆菌活菌攻击,比较各组胃粘膜的幽门螺杆菌定植密度,攻击前后三清中抗幽门螺杆菌抗体IgGI,IgG2a和IgA的变化,脾脏和胃粘膜中γ干扰素(IFN-γ)和白介素4(IL-4)mRNA表达变化及胃粘膜炎症情况。结果(1)KarA及Urea／Kara组的幽门螺杆菌定植密度显下降,Urea组下降不明显。(2)和生理盐水组相比攻击前各鼠伤寒沙门氏菌免疫组IgGI,IgG2a均轻度升高而IgA无变化,攻击后各鼠伤寒沙门氏菌免疫组IgG2a升高显,KatA和Urea／KatA组尤为明显,而IgGI和IgA的升高无统计学差异。(3)胃粘膜攻击前生理盐水组无IFN-γ表达,其余各组均100％表达;攻击后生理盐水组IFN-γ轻度表达但仍明显低于各鼹伤寒沙门氏菌免疫组。IL-4在攻击前后各组均无表达。(4)脾IFN-γ和IL-4在所有组攻击前后均全部表达。(5)攻击前各组鼠胃粘膜仅有散在少量白细胞,攻击后各鼠伤寒沙门氏菌免疫组出现明显以淋巴及单核细胞浸润为特征的炎症反应。结论以减毒鼠伤察沙门氏菌为载体构建的重组幽门螺杆菌疫苗(Katk、urek/kalk)可在小鼠体内诱导出以TH1反应为主并伴随免疫后胃炎均保护性免疫应答。     

幽门螺杆菌疫苗免疫小鼠的Th免疫应答及作用

目的 研究幽门螺杆菌(Helicobacter pylori,Hp)疫苗诱导小鼠保护性免疫应答的机制。方法 以霍乱毒素(CT)加Hp全菌超声粉碎抗原免疫小鼠，并设CT组和生理盐水(NS)组为对照。免疫4周后以Hp攻击；在攻击前后不同时间点分批处死小鼠，比较免疫组和对照组胃黏膜唧定植及各项免疫指标的变化。结果 (1)攻击后5、26周后免疫组和CT组、NS组相比胃黏膜Hp定植量明显降低。(2)各时间点免疫组IgG1、IgG2a、IgA均高于NS组、CT组。(3)攻击前和攻击后5周免疫组和CT组胃黏膜Th1型细胞因子表达较NS组明显升高。攻击后26周免疫组和CT组除白细胞介素(IL)-12外，干扰素(IFN)-γ和IL-2表达均明显降低，而NS组Thl型细胞因子表达明显升高。(4)Th2型细胞因子在攻击前免疫组和CT组胃黏膜均有轻度表达；攻击后5周免疫组和CT组IL-4和IL-10均无表达，IL-6表达低于NS组；攻击后26周免疫组IL-4和IL-10再次表达，CT组仅表达IL-10，IL-6表达各组无明显差异。(5)各细胞因子在所有组脾脏攻击前后各时间点表达无差异。(6)与NS组相比，攻击后5周免疫组鼠胃黏膜出现明显的炎症反应，CT组也有轻度炎症反应；攻击后26周NS组炎症加重而免疫组则有所减轻。结论 (1)以CT为佐剂的Hp疫苗可诱导Thl和Th2混合型初始免疫应答；(2)Hp攻击后早期增强的Thl反应起主要免疫保护作用同时导致免疫后胃炎；晚期随Th2反应的增强Thl反应和免疫后胃炎强度减弱。     

幽门螺杆菌疫苗口服免疫小鼠后胃肠免疫应答研究

目的 观测Hp全菌抗原和粘膜佐剂LT口服免疫Balb/c小鼠后的胃肠免疫反应。方法 采用ELISPOT和ELISA法检测了胃粘膜、PP结抗原特异性形成细胞(AFC)和小肠粘液sIgA。结果 抗原免疫组、抗原 佐剂组胃肠粘膜抗原特异性sIgA-AFC、IgG-AFC数量明显增加,尤以sIgA-AFC为甚,并且抗原 佐剂组明显高于单纯抗原组和对照组;小肠粘液特异sIgA两免疫组明显高于对照组。结论 口服免疫可有效诱导胃肠道粘膜免疫,局部sIgA可能在抗Hp感染中具有重要作用。     

幽门螺杆菌感染相关胃炎

幽门螺杆菌感染相关胃炎陈希陶幽门螺杆菌（Ｈｅｌｉｃｏｂａｃｔｅｒｐｙｌｏｒｉ，Ｈｐ）感染人体胃粘膜后，引起胃及十二指肠粘膜炎症。Ｈｐ感染引起的胃炎是发病率最高的消化道疾病，在世界某些地区及发展中国家的人群中发病率尤高。西方某些发达国家的５０岁以上人群...     

幽门螺杆菌感染胃粘膜抗体产生细胞的研究

利用免疫组织化学方法对幽门螺杆菌(HP)感染胃粘膜内IgA,IgG和IgM产生细胞进行了研究.结果表明,HP感染引起了胃粘膜内局部抗体产生细胞的增加.其中以IgG和IgM产生细胞增加更着.提示:HP感染时局部体液免疫反应增强,进一步为HP是引起慢性活动性胃炎的病原提供了依据.     

幽门螺杆菌性胃炎的动物模型

许多研究者认为,幽门螺杆菌(Helicobacter Py-lori,HP)是胃炎和消化性溃疡的一个致病因子,目前对HP 感染的认识有了很快的发展。但是,正如人类的许多疾病一样,如果能建立一个供实验用的HP 感染动物模型,将会使人们对HP相关性胃炎的认识有一个飞跃。为未来研究着想,应优先考虑在常规饲养动物中选择优良的动物作为感染的模型,这样一个动物模型的建立可以帮助完善Koch’s 原则,明确致病机理和治疗方法。     

幽门螺杆菌感染的免疫防治

回顾和总结了幽门螺杆蓖疫苗研制中的一些主要问题如免疫原及佐剂的选择,动物模型的建立,免疫途径及疫苗转送系统的确定以及免疫保护机制的探讨,并对今后的研究方向作了进一步展望。     

幽门螺杆菌感染BALB/c无菌小鼠的免疫应答

目的 探讨幽门螺杆菌感染步鼠的免疫就应答状况。方法 建立ＨＰ经口感染无菌小鼠的动物模型,设立感染免疫组、感染对照组与免疫对照组３组,分别进行试验。结果 感染免疫组抗ＨＰ的特异性ＩｇＧ有显著增高,且该组小鼠胃中的ＨＰ菌数大量减少。结论此种免疫应答以体液免疫为主,细胞免疫所占比例很少,不能彻底清除小鼠胃中的ＨＰ。     

幽门螺杆菌与胃炎

幽门螺杆菌与胃炎潘占荣贵州省凯里市黔东南州人民医院内科５５６０００ＳｕｂｊｅｃｔｈｅａｄｉｎｇｓＧａｓｔｒｉｔｉｓ／ｍｉｃｒｏｂｉｏｌｏｇｙＨｅｌｉｃｏｂａｃｔｅｒｉｎｆｅｃｔｉｏｎｓ／ｃｏｍｐｌｉｃａｔｉｏｎｓＨｅｌｉｃｏｂａｃｔｅｒｐｙｌｏｒ...     

慢性萎缩性胃炎免疫状态与幽门螺杆菌感染的关系

本文对１１０例老年人慢性萎缩性胃炎检测幽门螺杆菌及免疫球蛋白ＩｇＡ、ＩｇＧ，Ｔ细胞标民，和超氧化物歧化酶等局部粘膜免疫状态。结果显示老年ＣＡＧ的ＨＰ感染率比非老年ＣＡＧ的ＨＰ感染率为低。局部粘膜ＩｇＡ、ＩｇＧ阳性增强，且体液免疫ＩｇＡ、ＩｇＧ强于细胞免疫ＴｕｃＨＬ，有ＳＯＤ减弱现象。示感染ＨＰ的老年ＣＡＧ患者存在粘膜免疫功能亢进，以及清除自由基能力下降。     

胃窦部幽门螺杆菌清除对萎缩性胃炎病理改变的影响

目的通过对幽门螺杆菌（Hp）相关的萎缩性胃炎病人Hp清除治疗前后胃窦部黏膜病理改变的分析，来确定Hp对其炎症程度、活动性（中性粒细胞浸润）、腺体萎缩和肠上皮化生的影响。方法106例Hp相关的萎缩性胃炎患者接受Hp清除治疗，对其治疗前后胃窦部黏膜的病理变化进行分析，分析标准按96悉尼系统评定。结果在62例治疗成功组中，治疗后胃黏膜的炎症程度及活动性较治疗前明显减轻，但腺体萎缩及肠上皮化生未减轻。在44例治疗失败组中，治疗前后胃黏膜的炎症程度、活动性、腺体萎缩及肠化均没有变化。且随着Hp感染时间的延长，腺体萎缩和肠化还可加重。结论Hp的清除治疗可使萎缩性胃炎患者胃黏膜的炎症程度及活动度减轻，对此类病人应行Hp清除治疗。     

A short-term eradication therapy for Helicobacter pylori acute gastritis

BACKGROUND AND AIMS: Acute gastritis, caused by an initial infection of Helicobacter pylori (H. pylori), may resolve spontaneously, but the infection sometimes becomes chronic. We examined the efficacy of a short-term H. pylori eradication therapy on acute gastritis. METHODS: Among the 15 patients with hemorrhagic acute gastritis who were randomly allocated to group A (eradication therapy) or group B (Lansoprazole, LPZ), 10 of them started to receive treatment within 1 day after the disease onset. The other five patients began the eradication therapy 4-6 days after disease onset (group C). Eradication therapy consisted of a daily oral administration of each of 30 mg lansoprazole (LPZ), once a day; 400 mg clarithromycin, twice a day; 1000 mg amoxicillin, twice a day; and 300 mg rebamipide, three times a day, for one week. If the endoscopy was normal, medication was stopped for the following 4 weeks before gastric endoscopy was performed again in order to assess H. pylori eradication. RESULTS: All group A patients were cured after the 1-week treatment and therefore, they became H. pylori negative. Group B and C patients had erosions or ulcers after the 1-week treatment and so received an additional 3-week administration of LPZ. Four weeks later, their gastritis was cured and except for one group B patient, they became H. pylori-negative. CONCLUSION: In patients with acute gastritis, caused by an initial H. pylori infection, eradication therapy was efficacious in achieving early healing. This therapy should be started as soon as possible after disease onset.     

表达幽门螺杆菌尿素酶B亚单位的减毒鼠伤寒杆菌口服疫苗的制备

目的制备抗幽门螺杆菌(Hp)尿素酶B亚单位(UreB)减毒鼠伤寒杆菌活菌疫苗,观察其免疫效果.方法构建表达UreB的原核表达载体PTc01-UreB并转化减毒鼠伤寒杆菌SL3261,得到重组菌SL3261/PTc01-UreB.应用抗Hp菌体蛋白兔血清行Western-blot检测UreB在SL3261中的表达.将SL3261/PTc01-UreB口服免疫Balb/c小鼠,12周后应用ELISA检测肠液和血清中的特异性抗体反应.SL3261/PTc01-UreB在Luria-Bertani培养液中连续传代60代以确定其稳定性.结果成功构建PTc01-UreB原核表达载体.Western-blot显示,其转化减毒鼠伤寒杆菌SL3261后能表达相对分子质量约61×103的蛋白,与HpUreB亚单位相符,具有抗原性.口服免疫小鼠后,在肠液和血清中可分别检测到针对UreB的特异性IgA和IgG抗体.体外连续培养60代未见PTc01-UreB质粒丢失及对宿主细胞毒性.结论表达HpUreB的减毒鼠伤寒杆菌SL3261/PTc01-UreB可用作抗Hp感染口服疫苗.     

中西医结合根除幽门螺杆菌对萎缩性胃炎核因子-κB表达的影响

[目的]观察以胃舒散为主的三联疗法（胃舒散、呋喃唑酮和克拉霉素）治疗幽门螺杆菌（Hp）阳性慢性萎缩性胃炎（CAG）的临床效果及其对核因子-κB（NF-κB）表达的影响。[方法]41例Hp阳性CAG患者服用胃舒散2．0g，呋喃唑酮0．1g，各3次／d，克拉霉素0．25g，2次／d，1周后再继服胃舒散4周。治疗前及治疗结束1年后行内镜及病理组织学检查，取活检观察病理组织学改变及NF-κB表达变化，采用银染色法、^14C-尿素呼气试验或快速尿素酶试验检测Hp。[结果]三联疗法结束1年后，Hp根除率为85．4％；根除Hp能显著减轻患者胃窦部慢性炎症（P〈0．05）和活动程度（P〈0．01），下调NF-κB表达（P〈0．01），但胃炎的萎缩和肠化生等病理无明显改变。[结论]以胃舒散为主的三联疗法对Hp有较高根除率。根除Hp可抑制NF-κB的表达，减轻活动性炎症，但近期观察对萎缩、肠化生等病理改变无明显作用。     

Association of Helicobacter pylori infection with atrophic gastritis and intestinal metaplasia

AIMS: To evaluate the effect of Helicobacter pylori infection and aging on atrophy and intestinal metaplasia of the gastric mucosa. METHODS: One hundred and sixty-three patients were divided into three age groups and underwent an upper gastrointestinal endoscopy where no esophagitis, peptic ulcers, or malignancies were detected. Two biopsy specimens were obtained from the anterior and posterior walls of the antrum and of the fundus. These were used to evaluate the grade of gastritis, bacterial culture and histologic evidence of H. pylori infection. RESULTS: Helicobacter pylori infection was found to be directly associated with an increased risk of gastritis grade (odds ratio (OR) = 90 (95% CI; 30-270)). An age of 60 years and older along with H. pylori infection was also strongly associated with an increased risk of atrophy (OR = 6.6, (95% CI; 2.9-15.2)); OR = 9.8, (95% CI; 2.7-35.4)), as was intestinal metaplasia of the gastric mucosa (OR = 5.5, (95% CI; 1.7-17.6)); OR = 7.9, (95% CI; 2.8-46.1)). The prevalence of atrophic gastritis increased with advancing age in H. pylori-infected patients, but no such phenomenon was observed in H. pylori-uninfected patients. The prevalence of intestinal metaplasia significantly increased with advancing age, irrespective of the presence of H. pylori infection. In addition, H. pylori uninfected female patients had a decreased risk of intestinal metaplasia. CONCLUSIONS: These results suggest that atrophic gastritis is not a normal aging process, but instead is likely to be the result of H. pylori infection, while intestinal metaplasia is caused by both the aging process and H. pylori infection. A decreased risk of intestinal metaplasia found in uninfected female subjects may partly explain the lower prevalence of gastric cancer in females than in males.     

幽门螺杆菌感染对胃黏膜环氧合酶表达的影响

目的评估幽门螺杆菌（Hp）急、慢性感染对小鼠胃黏膜环氧合酶（COX）表达的影响.方法清洁级BALB/C小鼠共40只,分为阴性对照组（10只）予0.4ml生理盐水灌喂,急性感染组（10只）予0.4 ml Hp菌液灌喂,共5 d,该两组小鼠在最后一次灌喂后2周处死;另20只小鼠予0.4ml Hp菌液灌喂,共5d,在最后一次灌喂2个月后将小鼠均分为两组,治疗组予克拉霉素按每天13.5 mg/kg分2次灌喂,连续7d（本组有1只小鼠死亡）;慢性感染组予生理盐水,1个月后分别处死两组动物.取胃黏膜标本,用免疫组化半定量法测定上述各组胃窦黏膜COX-1及COX-2的蛋白表达.结果四组小鼠胃黏膜小凹上皮均可见COX-1蛋白表达,各组阳性率差异无统计学意义（P＞0.05）.与阴性对照组相比,急、慢性感染组COX-2蛋白表达均明显增加,其中慢性感染组治疗前COX-2阳性率明显高于急性感染组.慢性感染组治疗后阳性率与治疗前相比有所下降,但差异无统计学意义（P＞0.05）.结论Hp感染导致胃黏膜COX-2表达增加,对COX-1表达无明显作用,清除Hp后COX-2表达仍高于正常.     

幽门螺杆菌疫苗研究的现状和展望

幽门螺杆菌感染与慢性胃炎、消化性溃疡、胃癌及胃粘膜相关性淋巴瘤有关。疫苗是防治幽门螺杆菌感染切实可行的方法。近年来新的防治和根除幽门螺杆菌感染的疫苗正在开发并取得了很大的进展。本文对新近研制的蛋白组分疫苗、活菌疫苗、DNA疫苗、缓释微球疫苗及表位疫苗等幽门螺杆菌疫苗的进展作一综述。     

A topographic study of Helicobacter pylori density, distribution and associated gastritis

BACKGROUND AND AIMS: The topographic distribution and density of Helicobacter pylori and associated gastritis in the stomach were studied in order to determine which biopsy sites are likely to provide the maximum yield so as to reduce the fallacious results due to sampling error. METHODS: Fifty patients with upper gastrointestinal symptoms were studied. Eleven gastric biopsies from predetermined sites were obtained and subjected to ultra-rapid urease test, imprint cytology and histology. Haematoxylin and eosin stain was used for defining gastritis and other associated histopathological details. Loeffler's methylene blue stain was used to confirm the presence of H. pylori in imprint smears and histological sections. RESULTS: All 50 patients had H. pylori infection and evidence of chronic gastritis at one or more of the 11 biopsy sites. Maximum and minimum percentage positivity were observed at A3 (antral lesser curvature) and B4 (corpus greater curvature), respectively. Various sites in decreasing order of percentage positivity were A3 > A2 > A1 > A4 > B3 > B1 > A5 > B6 > B5 > B2 > B4. Among the biopsies obtained from the corpus, B3 (corpus lesser curvature) was the site with maximum positivity. A3 and B4 had a statistically significant difference in percentage positivity (P < 0.0001) for H. pylori and gastritis. The maximum and minimum density scores of H. pylori and gastritis were found in A3 and the B4, respectively. A3 had a significantly higher (P < 0.0001) mean density score than any other site in the stomach. The difference in the grading of H. pylori between A3 and B3 (sites of maximum positivity in antrum and corpus) was statistically significant (P < 0.0001). A statistically significant (P < 0.001) positive correlation between increasing grades of H. pylori and gastritis was observed at the site of maximum density. Eighty per cent of the patients had antral predominant gastritis and in 82%, H. pylori was predominantly observed in antral biopsies. CONCLUSION: It is concluded that two biopsies taken from A3 are sufficient for confirmation of presence of H. pylori and associated gastritis for initiation of treatment. However, additional biopsies from B3 will help in deciding the topographic pattern of gastritis.     

Treatment of Helicobacter pylori infection in atrophic gastritis

Helicobacter pylori(Hp) is a major human pathogen causing chronic, progressive gastric mucosal damage and is linked to gastric atrophy and cancer. Hp-positive individuals constitute the major reservoir for transmission of infection. There is no ideal treatment for Hp. Hp infection is not cured by a single antibiotic, and sometimes, a combined treatment with three or more antibiotics is ineffective. Atrophic gastritis(AG) is a chronic disease whose main features are atrophy and/or intestinal metaplasia of the gastric glands, which arise from long-standing Hp infection. AG is reportedly linked to an increased risk for gastric cancer, particularly when extensive intestinal metaplasia is present. Active or past Hp infection may be detected by conventional methods in about two-thirds of AG patients. By immunoblotting of sera against Hp whole-cell protein lysates, a previous exposure to Hp infection is detected in all AG patients. According to guidelines, AG patients with Hp positivity should receive eradication treatment. The goals of treatment are as follows:(1) Cure of infection, resolution of inflammation and normalization of gastric functions;(2) possible reversal of atrophic and metaplastic changes of the gastric mucosa; and(3) prevention of gastric cancer. An ideal antibiotic regimen for Hp should achieve eradication rates of approximately 90%, and complex multidrug regimens are required to reach this goal. Amongst the factors associated with treatment failure are high bacterial load, high gastric acidity, Hp strain, smoking, low compliance, overweight, and increasing antibiotic resistance. AG, when involving the corporal mucosa, is linked to reduced gastric acid secretion. At a non-acidic intra-gastric p H, the efficacy of the common treatment regimens combining proton pump inhibitors with one or more antibiotics may not be the same as that observed in patients with Hp gastritis in an acid-producing stomach. Although the efficacy of these therapeutic regimens has been thoroughly tested in subjects with Hp infection, there is a paucity of evidence in the subgroupof patients with AG. Bismuth-based therapy may be an attractive treatment in the specific setting of AG, and specific studies on the efficacy of bismuth-based therapies are needed in patients with AG.