未经授权:抗原特异性免疫疗法是治疗HBV感染和肝细胞癌(HCC)的有希望的策略。为了促进杀死恶性和/或感染的肝细胞,重要的是要知道哪些T细胞靶标是由人类白细胞抗原(HLA)-I复合物在患者来源的肝细胞上呈递的。这里,我们旨在揭示肝细胞特异性HLA-I肽组,重点是HBV蛋白和肿瘤相关抗原(TAA)衍生的肽,以指导抗原特异性免疫治疗的发展。
UNASSIGNED:使用新设计的无灌注程序从(HBV感染)非肿瘤和HCC组织中高纯度分离原代人肝细胞。通过无偏倚质谱(MS)鉴定肝细胞衍生的HLA结合肽,然后对源蛋白进行基因本体论和通路分析。使用靶向MS搜索HBV抗原和TAA衍生的HLA肽,并测试了一些肽的免疫原性。
未经评估:使用无偏数据相关采集(DDA),我们获得了2×105肽的高质量HLA-I肽组,该肽组包含8种HBV衍生肽和来自8种已知HCC相关TAA的14种肽,这些肽是肿瘤专有的.其中,在样品中通过靶向MS检测到3种HBV-和12种TAA衍生的HLA肽,它们最初通过DDA鉴定。此外,在未使用无偏倚MS进行鉴定的样品中检测到2种HBV和2种TAA衍生的HLA肽。最后,5个HBV衍生肽和3个TAA衍生肽证明了免疫原性。
未经授权:我们提出了分离的原代人肝细胞的第一个HLA-I免疫肽组,缺乏免疫细胞。鉴定的HBV衍生和TAA衍生的肽直接帮助开发用于慢性HBV感染和HCC的抗原特异性免疫疗法。所描述的方法还可以应用于一般肝脏疾病的个性化免疫治疗治疗。
UNASSIGNED:旨在诱导针对病毒或肿瘤的免疫反应的免疫治疗是治疗慢性HBV感染和肝癌的一种有前途的新型治疗选择。为了设计成功的治疗方法,重要的是要知道病毒来源和肿瘤特异性蛋白的哪些片段(即肽)通过患病的肝细胞呈递给免疫系统的T细胞,并且因此是免疫疗法的良好靶标。这里,我们从患有慢性HBV感染和/或肝癌的患者中分离出肝细胞,分析了这些细胞提供的肽,并评估哪些肽能够驱动免疫反应。
UNASSIGNED: Antigen-specific immunotherapy is a promising strategy to treat HBV infection and hepatocellular carcinoma (HCC). To facilitate killing of malignant and/or infected hepatocytes, it is vital to know which T cell targets are presented by human leucocyte antigen (HLA)-I complexes on patient-derived hepatocytes. Here, we aimed to reveal the hepatocyte-specific HLA-I peptidome with emphasis on peptides derived from HBV proteins and tumour-associated antigens (TAA) to guide development of antigen-specific immunotherapy.
UNASSIGNED: Primary human hepatocytes were isolated with high purity from (HBV-infected) non-tumour and HCC tissues using a newly designed perfusion-free procedure. Hepatocyte-derived HLA-bound peptides were identified by unbiased mass spectrometry (MS), after which source proteins were subjected to Gene Ontology and pathway analysis. HBV antigen and TAA-derived HLA peptides were searched for using targeted MS, and a selection of peptides was tested for immunogenicity.
UNASSIGNED: Using unbiased data-dependent acquisition (DDA), we acquired a high-quality HLA-I peptidome of 2 × 105 peptides that contained 8 HBV-derived peptides and 14 peptides from 8 known HCC-associated TAA that were exclusive to tumours. Of these, 3 HBV- and 12 TAA-derived HLA peptides were detected by targeted MS in the sample they were originally identified in by DDA. Moreover, 2 HBV- and 2 TAA-derived HLA peptides were detected in samples in which no identification was made using unbiased MS. Finally, immunogenicity was demonstrated for 5 HBV-derived and 3 TAA-derived peptides.
UNASSIGNED: We present a first HLA-I immunopeptidome of isolated primary human hepatocytes, devoid of immune cells. Identified HBV-derived and TAA-derived peptides directly aid development of antigen-specific immunotherapy for chronic HBV infection and HCC. The described methodology can also be applied to personalise immunotherapeutic treatment of liver diseases in general.
UNASSIGNED: Immunotherapy that aims to induce immune responses against a virus or tumour is a promising novel treatment option to treat chronic HBV infection and liver cancer. For the design of successful therapy, it is essential to know which fragments (i.e. peptides) of virus-derived and tumour-specific proteins are presented to the T cells of the immune system by diseased liver cells and are thus good targets for immunotherapy. Here, we have isolated liver cells from patients who have chronic HBV infection and/or liver cancer, analysed what peptides are presented by these cells, and assessed which peptides are able to drive immune responses.