Immunoelectron microscopic study on the expression of lymphoid markers in HTLV- producing cells

HTLV产生细胞淋巴标志物表达的免疫电镜研究

• 批准号: 10670209
• 负责人: OHTSUKI Yuji
• 金额: $ 1.6万
• 依托单位: Kochi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670209/
• 关键词: HTLV; T cell receptor; CD3; HHV-8; Ultrastructure; Primary effusion lymphoma; HHV-8; PEL

The T-cell receptor (TCR)/CD3 complex on the surface or T-lymphcytes is required for recognition of the antigens presented by macrophages. Usually, such antigens are presented on Major Histocompatibility Complex (MHC) molecules. In this recognition system, extracellular domains of both TCR α and β chains are reported to be essential for antigen recognition.On the other hand, it has been reported that the antigenicities of HTLV-I and -II producers, which are initially T-cells, change during cultivation, for example loosing some T-cell antigens or obtaining HLA-DR and/or CD25 antigens, as also reported in cases of high grade T-cell malignancy.In the present study, immunoelectron microscopic observation was performed to clarify TCR/CD3 complex expression on the surface of human T-lymphotropic virus (HTLV) type I- and type II-producing cells. Staining with WT-31 (TCR α/β) was found to be positive only in PM, while staining for αF1 (TCR α) and for βF1 (TCR β) was positive only in NM and rER … More . CD3, on the other hand, reacts with both NM/rER and PM.Using these antibodies, normal fresh, and fresh and short-term cultured HTLV-I carrier lymphocytes were clearly shown to possess TCR/CD3 complexes, lacking TCR α/β complex in particular, suggesting that their recognition of antigens is defcctive.The ultrastructure of Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus-8 (HHV-8) has not yet been fully elucidated, although some findings have been reported using primary effusion lymphoma (PEL) cell lines, KS-1, harboring no Epstein-Barr virus (EBV) coinfection. In the present study, detailed fine structural examination of KSHV/HHV-8 was performed after stimulation of the PEL-derived cell line KS-1 with 12-O-tetradecanoy1-phorbol-13-acetate (TPA) in vitro. While nuclear virus particles associated with no extracellular mature particles. KS-1 cells stimulated with TPA produced many extracellular mature particles as well as intranuclear particles, in addition to interesting tubulo-reticular structures and aggregated tubular structures in vesicles. The induced intranuclear particles were empty, doughnut shaped, and dense cored, with outer, and inner diameters of 100-110nm and 60-70nm, respectively. Dense-cored extracellular mature particles were 150-160nm in diameter, and some contained doughnut-shaped cores, together with a few megaloviruses, 260nm in outer diameter. These findings indicate that KS-1 cells treated with TPA can produce extracellular mature particles as well as intranuclear particles, which were proven to be KSHV/HHV-8. Less

巨噬细胞表面或T淋巴细胞表面的T细胞受体(TCR)/CD3复合体是识别巨噬细胞呈递的抗原所必需的。通常，这种抗原存在于主要组织相容性复合体(MHC)分子上。在这个识别系统中，TCRCD3链的胞外区被认为是抗原识别所必需的。另一方面，据报道，在培养过程中，最初为T细胞的α和β产生者的抗原性发生了变化，例如失去了一些T细胞抗原或获得了HLADR和/或CD25型抗原，这在高度恶性T细胞肿瘤中也有报道。WT-31(TcRα/β)染色仅在PM中阳性，而αF1(Tcrα)和βF1(Tcrβ)仅在NM和RER…中阳性更多。使用这些抗体，正常新鲜、新鲜和短期培养的HTLV-I载体淋巴细胞明显地具有TcR/CD3复合体，特别是缺乏TcRα/β复合体，这表明它们对抗原的识别存在缺陷。卡波西肉瘤相关疱疹病毒/人类疱疹病毒8(HV-8)的超微结构尚未完全阐明，尽管已有报道使用原代渗出性淋巴瘤细胞系KS-1，没有EB病毒的混合感染。本研究用12-O-十四烷酰-1-佛波醇-13-乙酸酯(TPA)体外刺激KSHV/HHV-8细胞株KS-1，对KSHV/HHV-8细胞进行精细结构观察。而核病毒颗粒与无细胞外成熟颗粒有关。经TPA刺激的KS-1细胞在胞外和核内产生许多成熟颗粒，并在囊泡中形成有趣的小管-网状结构和聚集的小管结构。诱导后的核内颗粒为空的、环状的、核心致密的，外径为100~110 nm，内径为60~70 nm。致密的胞外成熟颗粒直径150-160 nm，有的含有外径260 nm的甜甜圈状核心和少量巨型病毒。这些结果表明，经TPA处理的KS-1细胞能够产生成熟的胞外颗粒和胞内颗粒，证实为KSHV/HHV-8。较少

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