Kawakubo Masatomo, Horiuchi Kazuki, Komura Hitomi, Sato Yoshiko, Kato Masayoshi, Ikeyama Meguru, Fukushima Mana, Yamada Shigenori, Ishizone Satoshi, Matsumoto Takehisa, Ota Hiroyoshi, Sagara Junji, Nakayama Jun
Department of Molecular Pathology, Shinshu University Graduate School of Medicine, Matsumoto, 390-8621, Japan.
Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, 390-8621, Japan.
Histochem Cell Biol. 2017 Oct;148(4):463-471. doi: 10.1007/s00418-017-1582-4. Epub 2017 May 22.
Helicobacter suis (H. suis), formerly called Helicobacter heilmannii type 1 (H. heilmannii), is a gram-negative bacterium of the Helicobacter species. This pathogen infects the stomach of humans and animals such as dogs, cats, pigs, and rodents, the latter giving rise to zoonotic infection. Here, we generated a H. suis-specific antibody useful for immunohistochemistry with formalin-fixed, paraffin-embedded tissue sections. To do so, we began by cloning the gene encoding H. suis cholesterol α-glucosyltransferase (αCgT). αCgT is the key enzyme responsible for biosynthesis of cholesteryl α-D-glucopyranoside (CGL), a major cell wall component of Helicobacter species including H. suis. The deduced amino acid sequence of H. suis αCgT had 56% identity with the corresponding Helicobacter pylori (H. pylori). We then developed a polyclonal antibody (anti-Hh-I205R) by immunizing rabbits with a 205 amino acid H. suis αCgT fragment. Immunohistochemistry with the anti-Hh-I205R antibody could differentiate H. suis from H. pylori in gastric mucosa sections derived from mice infected with either pathogen. We then probed formalin-fixed, paraffin-embedded sections of human gastric mucosa positive for H. suis infection with the anti-Hh-I205R antibody and detected positive staining. These results indicate that anti-Hh-I205R antibody is specific for H. suis αCgT and useful to detect H. suis in gastric specimens routinely analyzed in pathological examinations.
猪幽门螺杆菌（Helicobacter suis，H. suis），以前称为海尔曼幽门螺杆菌1型（Helicobacter heilmannii type 1，H. heilmannii），是幽门螺杆菌属的一种革兰氏阴性菌。这种病原体感染人类以及狗、猫、猪和啮齿动物等动物的胃部，后者会引发人畜共患病感染。在此，我们制备了一种对福尔马林固定、石蜡包埋组织切片进行免疫组织化学有用的猪幽门螺杆菌特异性抗体。为此，我们首先克隆了编码猪幽门螺杆菌胆固醇α - 葡萄糖基转移酶（αCgT）的基因。αCgT是负责合成胆固醇α - D - 吡喃葡萄糖苷（CGL）的关键酶，CGL是包括猪幽门螺杆菌在内的幽门螺杆菌属的主要细胞壁成分。猪幽门螺杆菌αCgT的推导氨基酸序列与相应的幽门螺杆菌（Helicobacter pylori，H. pylori）具有56%的同一性。然后，我们用一段205个氨基酸的猪幽门螺杆菌αCgT片段免疫兔子，制备了一种多克隆抗体（抗Hh - I205R）。用抗Hh - I205R抗体进行免疫组织化学可以在感染了这两种病原体之一的小鼠的胃黏膜切片中区分猪幽门螺杆菌和幽门螺杆菌。然后，我们用抗Hh - I205R抗体探测猪幽门螺杆菌感染呈阳性的人胃黏膜的福尔马林固定、石蜡包埋切片，并检测到阳性染色。这些结果表明抗Hh - I205R抗体对猪幽门螺杆菌αCgT具有特异性，并且可用于在病理检查中常规分析的胃标本中检测猪幽门螺杆菌。
