AN INVITRO ADHERENCE ASSAY REVEALS THAT HELICOBACTER-PYLORI EXHIBITS CELL LINEAGE-SPECIFIC TROPISM IN THE HUMAN GASTRIC EPITHELIUM
AN INVITRO ADHERENCE ASSAY REVEALS THAT HELICOBACTER-PYLORI EXHIBITS CELL LINEAGE-SPECIFIC TROPISM IN THE HUMAN GASTRIC EPITHELIUM
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DOI：
10.1073/pnas.90.5.2035
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发表时间：
1993-03-01
影响因子：
11.1
通讯作者：
NORMARK, S
中科院分区：
文献类型：
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作者：
FALK, P;ROTH, KA;NORMARK, S
幽门螺杆菌是一种微需氧菌，存在于无症状人群以及酸性消化道疾病和胃腺癌患者的胃中。我们已经开发了一种原位粘附试验，以检查这种生物体的细胞系特异性结合的性质，并表征识别其粘附素的细胞表面受体的性质。异硫氰酸黄绿素标记的H.幽门螺杆菌菌株与人和大鼠胃单位小凹区的表面粘液细胞结合，但不与这些单位腺域中的粘液颈、壁或主细胞谱系结合。通过蛋白酶K处理组织切片和用牛颌下腺粘蛋白（岩藻糖基化和唾液酸化碳水化合物的丰富来源）预处理细菌，废除结合。几条证据表明，与表面粘液细胞的结合不依赖于粘附素受体中末端未取代的α 2，3-和α 2，6-连接的唾液酸：（i）孵育H.幽门螺杆菌菌株与唾液酸化的糖缀合物，如胎球蛋白和游离唾液酸乳糖;（ii）免疫组化染色使用唾液酸特异性接骨木和Maackia amurensis凝集素和霍乱毒素B亚单位没有检测到任何唾液酸化的糖缀合物在这些上皮细胞;和（iii）结合是不敏感的条件下，选择性裂解碳8和9的末端未修饰的唾液酸偏高碘酸盐。糖蛋白中岩藻糖基化表位介导H. pylori与表面粘液细胞结合的事实表明，该谱系共表达粘附素受体和主要岩藻糖基化组织血型抗原，对组织血型抗原H、B和Le（B）特异的单克隆抗体阻断结合，并且对α-L-岩藻糖特异的1型荆豆凝集素也与结合细菌的相同细胞结合。此外，人初乳分泌型伊加以偏高碘酸盐和α-L-岩藻糖苷酶敏感但不依赖于神经氨酸酶的方式抑制粘附。原位粘附试验可用于进一步鉴定H。幽门螺杆菌粘附素及其受体，以及用于鉴定抑制该人类病原体的菌株特异性和细胞系特异性结合的治疗上有用的化合物。
Helicobacter pylori is a microaerophilic bacterium found in the stomach of asymptomatic humans as well as patients with acid peptic disease and gastric adenocarcinoma. We have developed an in situ adherence assay to examine the cell lineage-specific nature of binding of this organism and to characterize the nature of cell surface receptors that recognize its adhesin. Fluorescein isothiocyanate-labeled H. pylori strains were bound to surface mucous cells present in the pit region of human and rat gastric units but not to mucous neck, parietal, or chief cell lineages present in the glandular domains of these units. Binding was abolished by proteinase K treatment of tissue sections and by pretreatment of the bacteria with bovine submaxillary gland mucin, a rich source of fucosylated and sialylated carbohydrates. Several lines of evidence suggest that binding to surface mucous cells is not dependent upon terminal nonsubstituted alpha2,3- and alpha2,6-linked sialic acids in the adhesin receptor: (i) binding was not inhibited by incubating H. pylori strains with sialylated glycoconjugates such as fetuin and free sialyllactose; (ii) immunohistochemical stainings using the sialic acid-specific Sambucus nigra and Maackia amurensis lectins and the cholera toxin B subunit did not detect any sialylated glycoconjugates in these epithelial cells; and (iii) binding was not sensitive to metaperiodate under conditions that selectively cleaved carbons 8 and 9 of terminal nonmodified sialic acids. A role for fucosylated epitopes in the glycoprotein(s) that mediate binding of H. pylori to surface mucous cells was suggested by the facts that this lineage coexpresses the adhesin receptor and major fucosylated histo-blood group antigens, that monoclonal antibodies specific for histo-blood group antigens H, B, and Le(b) block binding, and that the lectin Ulex europaeus type 1 agglutinin, which is specific for alpha-L-fucose, also bound to the same cells that bound the bacteria. Furthermore, human colostrum secretory IgA inhibited adhesion in a metaperiodate- and alpha-L-fucosidase-sensitive but neuraminidase-independent fashion. The in situ adherence assay should be useful in further characterizing the H. pylori adhesin and its receptor and for identifying therapeutically useful compounds that inhibit strain-specific and cell lineage-specific binding of this human pathogen.