DETERMINATION OF STRUCTURE OF AMYLOIDOGENIC IMMUNOGLOBULIN LIGHT CHAIN AND APPLICATION TO MOLECULAR DIAGNOSIS

促淀粉样免疫球蛋白轻链结构的测定及其在分子诊断中的应用

• 批准号: 07670200
• 负责人: MIURA Katsutoshi
• 金额: $ 1.41万
• 依托单位: HAMAMATSU UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670200/
• 关键词: AMYLOID; IMMUNOGLOBULIN; POLYNEUROPATHY; INFLAMMATION; HUMAN PAPILLOMA VIRUS; PCR; IN SITU HYBRIDIZATION; 扁桃; アミロイドーシス; エドマン分解; PCR反応

Amyloid fibrils were extracted in water and dissolved in 7M urea. Main paptide was separated with SDS polyacrylamide gel electrophoresis and transfered on PVDF membrane. Main peptides were cut from the membrane and subjected to N-terminal amino acid sequence analysis. A pulmonary nodular amyloidosis with systemic involvement was determined as lambda III type AL-amyloidosis, while a tonsillar amyloidosis of localized type was lambda VI type AL-amyloidosis.A case of familial amyloidosis with polyneuropathy had amyloid fibrils composed of transthyretin variant with substitution leucine for valine at position 30.Acute phase proteins such as SAA are produced in liver induced by cytokines from inflammatory sites and enter into the blood circulation. They reach the inflammatory sites, act against tissue damage, and suppress the inflammation. I reviewed this system and summarized in a chapter of a book.To investigate a association of HPV with tumorgenesis in unterine cervix and head and neck region, we extracted DNA from tumors, amplyfied HPV genome with PCR method, and detected the localization of HPV genome in the tumor by in situ hybridazation (ISH). In malignant lesions of uterine cervix, HPV genomes of malignant type were detected in about 90% most of which were not detected in ISH.In head and neck, esophageal squamous cell carcinoma (SCC) and tonsillar SCC contained HPV52b and HPV 16 respectively.We reported cases of papillary adenocarcinoma of mid-ear, metastatic sebacious carcinoma to the parotid gland, hyalinizing clear cell carcinoma of minor salivcary gland which recurred several times in 13 years, subglottal squamous cell carcinoma with hematogenous metastatis to thyroid, and verrucous carcinoma of temporal bone.

在水中提取淀粉样原纤维并溶解于7 M尿素中。用SDS聚丙烯酰胺凝胶电泳分离主肽，并转移到聚偏氟乙烯膜上。从膜上切下主要肽，并进行N-末端氨基酸序列分析。伴有全身受累的肺结节性淀粉样变性被确定为λ III型AL-淀粉样变性，而局限型扁桃体淀粉样变性为λ VI型AL，一个家族性淀粉样变性伴多发性神经病的病例具有由转甲状腺素蛋白变体组成的淀粉样纤维，其30位的缬氨酸被亮氨酸取代。血液循环。它们到达炎症部位，对抗组织损伤并抑制炎症。为了探讨HPV与宫颈及头颈部肿瘤发生的关系，我们从宫颈及头颈部肿瘤组织中提取DNA，用PCR方法扩增HPV基因组，用原位杂交技术（ISH）检测HPV基因组在宫颈及头颈部肿瘤组织中的定位。在宫颈恶性病变中，约90%的病例检出了HPV恶性型基因组，其中绝大多数在原位杂交中未检出，在头颈部，食管鳞状细胞癌（SCC）和扁桃体SCC中分别检出了HPV 52 b和HPV 16。13年内多次复发的小涎腺透明细胞癌、声门下鳞状细胞癌伴甲状腺血行转移和颞骨疣状癌。

项目成果

MIURA K.: Inflammation and infection. Acute phase protein.Current encyclopedia of pathology 5, Nakayama-shotten, 207-216 (1995)

MIURA K.：炎症和感染。

三浦克敏: "現代病理学大系5、炎症と感染、急性期蛋白" 中山書店, 428 (1995)

三浦胜俊：《现代病理学 5，炎症、感染和急性期蛋白》中山书店，428 (1995)

Shimizu H,Ishikawa K,Kobayashi H,Murakami T,Nakasato M,Miura K,Atsumi T :"An autopsy case of familial amyloidotic polyneuropathy due to deposition of variant transthyretin (va130*Leu)." Noshinnkei. 48(2). 175-178 (1996)

Shimizu H、Ishikawa K、Kobayashi H、Murakami T、Nakasato M、Miura K、Atsumi T：“由于变异转甲状腺素蛋白 (va130*Leu) 沉积导致的家族性淀粉样多发性神经病的尸检病例。”

Miura K :"Hyalinizing clear cell carcinoma of minor salivary gland showing repeated recurrence in 13 years." Byoinn byori. 13(2). 140 (1996)

Miura K：“小唾液腺透明细胞癌在 13 年内反复复发。”

Harada H,Kawabata Y,Nakajima A,Miura K :"Detection of HPV genome in uterine cervical lesions using in situ hybridization." Nippon rinnshou saibou gakkai kaishi. 34(2). 243 (1995)

Harada H、Kawabata Y、Nakajima A、Miura K：“利用原位杂交检测宫颈病变中的 HPV 基因组。”