Cloning of Helicobacter suis cholesterol α-glucosyltransferase and production of an antibody capable of detecting it in formalin-fixed, paraffin-embedded gastric tissue sections,
Cloning of Helicobacter suis cholesterol α-glucosyltransferase and production of an antibody capable of detecting it in formalin-fixed, paraffin-embedded gastric tissue sections,
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克隆猪螺杆菌胆固醇α-葡萄糖基转移酶并产生能够在福尔马林固定、石蜡包埋的胃组织切片中检测到该酶的抗体，
DOI：
10.1007/s00418-017-1582-4
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发表时间：
2017
影响因子：
2.3
通讯作者：
Jun Nakayama
中科院分区：
文献类型：
--
作者：
Masatomo Kawakubo;Kazuki Horiuchi;Hitomi Komura;Yoshiko Sato;Masayoshi Kato;Meguru Ikeyama;Mana Fukushima;Shigenori Yamada;Satoshi Ishizone;Takehisa Matsumoto;Hiroyoshi Ota;Junji Sagara;Jun Nakayama
猪螺杆菌（H. suis），以前称为Helicobacter heilmanniitype1（H. heilmannii）是螺旋杆菌属的革兰氏阴性细菌。这种病原体感染人类和动物如狗、猫、猪和啮齿动物的胃，后者引起人畜共患感染。在这里，我们生成了aH。用于福尔马林固定石蜡包埋组织切片免疫组织化学的猪特异性抗体。为此，我们首先克隆了编码H的基因。猪胆固醇α-葡萄糖基转移酶（α CgT）。α-CgT是螺旋杆菌（Helicobacterium）细胞壁主要成分胆固醇基α-D-吡喃葡萄糖苷（CGL）生物合成的关键酶。猪。推导的H. suis α CgT与幽门螺杆菌（H. pylori，H. pylori）。用205个氨基酸的Hh-I205R免疫家兔，制备了多克隆抗体（抗Hh-I205R）。猪α CgT片段。Hh-I205R抗体的免疫组化可区分H. suisfromH.幽门螺杆菌在来自感染任一病原体的小鼠的胃粘膜切片中。然后，我们探测福尔马林固定，石蜡包埋切片的人胃粘膜阳性H。用抗Hh-I205R抗体感染猪，检测到阳性染色。这些结果表明抗Hh-I205R抗体对H具有特异性。suis α CgT，对H.在病理检查中常规分析猪胃标本。
Helicobacter suis(H. suis), formerly calledHelicobacter heilmanniitype 1 (H. heilmannii), is a gram-negative bacterium of theHelicobacterspecies. 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 aH. suis-specific antibody useful for immunohistochemistry with formalin-fixed, paraffin-embedded tissue sections. To do so, we began by cloning the gene encodingH. suischolesterol α-glucosyltransferase (αCgT). αCgT is the key enzyme responsible for biosynthesis of cholesteryl α-d-glucopyranoside (CGL), a major cell wall component ofHelicobacterspecies includingH. suis. The deduced amino acid sequence ofH. suisαCgT had 56% identity with the correspondingHelicobacter pylori(H. pylori). We then developed a polyclonal antibody (anti-Hh-I205R) by immunizing rabbits with a 205 amino acidH. suisαCgT fragment. Immunohistochemistry with the anti-Hh-I205R antibody could differentiateH. suisfromH. pyloriin gastric mucosa sections derived from mice infected with either pathogen. We then probed formalin-fixed, paraffin-embedded sections of human gastric mucosa positive forH. suisinfection with the anti-Hh-I205R antibody and detected positive staining. These results indicate that anti-Hh-I205R antibody is specific forH. suisαCgT and useful to detectH. suisin gastric specimens routinely analyzed in pathological examinations.