Krakowka S, Ringler S S, Eaton K A, Green W B, Leunk R
Department of Veterinary Biosciences, College of Veterinary Medicine, Ohio State University, Columbus 43210, USA.
Vet Immunol Immunopathol. 1996 Jul;52(3):159-73. doi: 10.1016/0165-2427(95)05547-9.
Helicobacter pylori, a human gastric bacterial pathogen, was inoculated into gnotobiotic piglets and manifestations of the resultant gastric inflammation was analyzed by in situ immunochemistry and flow cytometric analysis of isolated lamina propria leukocytes (LPL) and peripheral blood leukocytes (PBL) recovered from infected and control piglets. Gastric mucosa tissue sections from uninfected control piglets were essentially negative for cluster differentiation- (CD-) positive leukocytes. Failure to isolate significant numbers of LPL from the gastric lamina propria confirmed this observation. A local and systemic immune response occurs in piglets after infection with H. pylori. This is manifest by the appearance of cells associated with a local immune response in gastric mucosa. In gastric tissue sections from H. pylori-infected piglets, CD4-positive leukocytes were sparse and closely associated with developing lymphoid follicles whereas the CD8-positive cellular phenotype was abundant. The latter formed a continuous band in the lamina propria just above the muscularis mucosa. Perivascular accumulations of lymphocytes in the outer muscular tunic(s) were strongly positive for expression of CD8 antigen. Class II-positive cells were prominent in CD8 lymphocytic infiltrates, developing follicles and vascular endothelia but were uniformly absent from gastric epithelia even in sites overlying areas of immunocyte proliferation and infiltration. Leukocytes possessing the monocyte and granulocyte markers were rare. Plasma cells containing IgA were common in the periphery of developing lymphoid follicles or distributed as discrete foci around individual gastric pits. Fewer numbers of IgG- and IgM-positive plasma cells were identified. When the LPL flow cytometry data were compared with the flow cytometry data obtained from PBL in these same H. pylori-infected piglets, leukocytes bearing the CD8 marker predominated in LPL whereas leukocytes bearing the CD4-reactive and MHC class II markers predominated in PBL. Finally, local ELISA antibody responses were measured in mucosal explant culture supernatants and compared with in vivo antibody levels in sera, bile, and gastric juice. Antibody activity, specific for H. pylori, was detected in supermatants and serum in all three isotypes in actively infected piglets whereas gastric juice lacked antibodies. Gastric explants prepared from piglets in which infection had been successfully eradicated failed to produce local antibody into supermatant fluids. These data support the concept that the gastric inflammation observed is mediated by local immunological events.
将人类胃部细菌病原体幽门螺杆菌接种到无菌仔猪体内，并通过原位免疫化学以及对从感染和对照仔猪中分离出的固有层白细胞（LPL）和外周血白细胞（PBL）进行流式细胞术分析，来分析由此产生的胃部炎症表现。未感染对照仔猪的胃黏膜组织切片中，分化簇（CD）阳性白细胞基本呈阴性。未能从胃固有层中分离出大量LPL证实了这一观察结果。幽门螺杆菌感染仔猪后会发生局部和全身免疫反应。这表现为胃黏膜中出现与局部免疫反应相关的细胞。在幽门螺杆菌感染仔猪的胃组织切片中，CD4阳性白细胞稀少且与发育中的淋巴滤泡紧密相关，而CD8阳性细胞表型丰富。后者在黏膜肌层上方的固有层中形成一条连续带。外层肌层中血管周围的淋巴细胞聚集对CD8抗原表达呈强阳性。II类阳性细胞在CD8淋巴细胞浸润、发育中的滤泡和血管内皮中很突出，但即使在免疫细胞增殖和浸润区域上方的部位，胃上皮中也均未出现。具有单核细胞和粒细胞标志物的白细胞很少见。含有IgA的浆细胞在发育中的淋巴滤泡周边很常见，或作为离散灶分布在各个胃小凹周围。鉴定出的IgG和IgM阳性浆细胞数量较少。当将这些相同的幽门螺杆菌感染仔猪的LPL流式细胞术数据与从PBL获得的流式细胞术数据进行比较时，带有CD8标志物的白细胞在LPL中占主导地位，而带有CD4反应性和MHC II类标志物的白细胞在PBL中占主导地位。最后，在黏膜外植体培养上清液中测量局部ELISA抗体反应，并与血清、胆汁和胃液中的体内抗体水平进行比较。在活跃感染仔猪的上清液和血清中，检测到了针对幽门螺杆菌的所有三种亚型的抗体活性，而胃液中缺乏抗体。从已成功根除感染的仔猪制备的胃外植体未能向上清液中产生局部抗体。这些数据支持了所观察到的胃部炎症是由局部免疫事件介导的这一概念。
