Conjugation of Latent Membrane Protein (LMP)-2 Epitope to Gold Nanoparticles as Highly Immunogenic Multiple Antigenic Peptides for Induction of Epstein-Barr Virus-Specific Cytotoxic T-Lymphocyte Responses in Vitro
Conjugation of Latent Membrane Protein (LMP)-2 Epitope to Gold Nanoparticles as Highly Immunogenic Multiple Antigenic Peptides for Induction of Epstein-Barr Virus-Specific Cytotoxic T-Lymphocyte Responses in Vitro
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DOI：
10.1021/bc800167q
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发表时间：
2009-01-01
影响因子：
4.7
通讯作者：
Yu, Wing-Yiu
中科院分区：
文献类型：
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作者：
Cheung, Wai-Hung;Chan, Vera Sau-Fong;Yu, Wing-Yiu
鼻咽癌是东南亚地区高发的一种肿瘤，它与eb病毒（EBV）激活密切相关，涉及一种弱免疫原性蛋白，即潜伏膜蛋白(LMP)-2的表达。先前的免疫学研究已经在LMP-2抗原中发现了人白细胞抗原(HLA)-A11限制性肽表位（SSCSSCPLSK）。在这项工作中，我们用n -半胱氨酸LMP-2表位处理纳米颗粒制备了金纳米颗粒(AuNP)-肽偶联物1。通过TEM（直径15 ~ 24 nm）和紫外-可见光谱（λ (max) = 520 nm处表面等离子共振吸收带）对aunp -肽偶联物进行了表征。在CALNN盖层肽存在的情况下，aunp -肽偶联物在室温下在溶液中稳定而不聚集至少48 In。通过ELIspot研究发现，与未偶联的LMP-2肽（SFC = 73 +/- 28）相比，aunp -肽偶联1在健康HLA-A11供者外周血单个核细胞中引起的nf - γ反应[斑点形成细胞数(SPC) = 727 +/- 198]明显更强。进一步的研究表明，用偶联物I处理树突状细胞可以在体外影响CD8+ T细胞活化，导致表位特异性细胞毒性T淋巴细胞杀伤反应。
Nasopharyngeal carcinoma is a neoplasm with a high incidence in Southeast Asia, and it is strongly associated with Epstein-Barr virus (EBV) activation involving the expression of a weakly immunogenic protein, namely, latent membrane protein (LMP)-2. Previous immunological studies already identified the human leukocyte antigen (HLA)-A11 restricted peptide epitope (SSCSSCPLSK) in the LMP-2 antigen. In this work, we prepared gold nanoparticle (AuNP)-peptide conjugate 1 by treating the nanoparticles with the N-cysteinated LMP-2 epitope. The AuNP-peptide conjugates have been characterized by TEM (15-24 nm in diameter) and UV-vis spectroscopy (surface plasmon resonance absorption band at lambda(max) = 520 nm). In the presence of a CALNN capping peptide, the AuNP-peptide conjugates are stable in solution without aggregation at room temperature for at least 48 In. By ELIspot studies, AuNP-peptide conjugate 1 was found to elicit a significantly stronger INF-gamma response [number of spot forming cells (SPC) = 727 +/- 198] from peripheral blood mononuclear cells of healthy HLA-A11 donors when compared to that induced by the unconjugated LMP-2 peptides (SFC = 73 +/- 28). Further studies showed that dendritic cells treated with conjugate I can effect CD8+ T-cell activation leading to epitope-specific cytotoxic T lymphocyte killing responses in vitro.