Onouchi Takanori, Mizutani Yasuyoshi, Shiogama Kazuya, Inada Ken-ichi, Okada Tatsuyoshi, Naito Kensei, Tsutsumi Yutaka
Department of Pathology, Fujita Health University School of Medicine, Toyoake, Japan.
Microbiol Immunol. 2015 Jan;59(1):13-27. doi: 10.1111/1348-0421.12213.
Streptococcus pyogenes is the main causative pathogen of recurrent tonsillitis. Histologically, lesions of recurrent tonsillitis contain numerous plasma cells. Strep A is an antigenic carbohydrate molecule on the cell wall of S. pyogenes. As expected, plasma cells in subjects with recurrent tonsillitis secrete antibodies against Strep A. The enzyme-labeled antigen method is a novel histochemical technique that visualizes specific antibody-producing cells in tissue sections by employing a biotin-labeled antigen as a probe. The purpose of the present study was to visualize plasma cells producing antibodies reactive with Strep A in recurrent tonsillitis. Firstly, the lymph nodes of rats immunized with boiled S. pyogenes were paraformaldehyde-fixed and specific plasma cells localized in frozen sections with biotinylated Strep A. Secondly, an enzyme-labeled antigen method was used on human tonsil surgically removed from 12 patients with recurrent tonsillitis. S. pyogenes genomes were PCR-detected in all 12 specimens. The emm genotypes belonged to emm12 in nine specimens and emm1 in three. Plasma cells producing anti-Strep A antibodies were demonstrated in prefixed frozen sections of rat lymph nodes, 8/12 human specimens from patients with recurrent tonsillitis but not in two control tonsils. In human tonsils, Strep A-reactive plasma cells were observed within the reticular squamous mucosa and just below the mucosa, and the specific antibodies belonged to either IgA or IgG classes. Our technique is effective in visualizing immunocytes producing specific antibodies against the bacterial carbohydrate antigen, and is thus a novel histochemical tool for analyzing immune reactions in infectious disorders.
化脓性链球菌是复发性扁桃体炎的主要致病病原体。从组织学上看，复发性扁桃体炎的病变中含有大量浆细胞。A群链球菌是化脓性链球菌细胞壁上的一种抗原性碳水化合物分子。不出所料，复发性扁桃体炎患者的浆细胞会分泌抗A群链球菌的抗体。酶标抗原法是一种新型组织化学技术，通过使用生物素标记的抗原作为探针，在组织切片中可视化产生特异性抗体的细胞。本研究的目的是在复发性扁桃体炎中可视化产生与A群链球菌反应性抗体的浆细胞。首先，用煮沸的化脓性链球菌免疫的大鼠淋巴结用多聚甲醛固定，并用生物素化的A群链球菌在冰冻切片中定位特异性浆细胞。其次，对12例复发性扁桃体炎患者手术切除的人扁桃体采用酶标抗原法。在所有12个标本中均通过PCR检测到化脓性链球菌基因组。9个标本的emm基因型属于emm12，3个属于emm1。在大鼠淋巴结的预固定冰冻切片、12例复发性扁桃体炎患者的8/12份人标本中证实了产生抗A群链球菌抗体的浆细胞，但在两个对照扁桃体中未发现。在人扁桃体中，在网状鳞状黏膜内和黏膜下方观察到与A群链球菌反应的浆细胞，特异性抗体属于IgA或IgG类。我们的技术可有效可视化产生针对细菌碳水化合物抗原的特异性抗体的免疫细胞，因此是一种用于分析感染性疾病免疫反应的新型组织化学工具。
