Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore (C.K.T., W.N.C., D.E.A., L.-F.W.); Singhealth Duke-NUS Global Health Institute, Singapore (C.K.T., L.-F.W.).
Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore (C.K.T., W.N.C., D.E.A., L.-F.W.).
Am J Ophthalmol. 2024 Oct;266:218-226. doi: 10.1016/j.ajo.2024.05.004. Epub 2024 May 21.
The identification of infectious etiologies is important in the management of uveitis. Ocular fluid testing is required, but multiplex testing faces challenges due to the limited volume sampled. The determination of antibody repertoire of aqueous humor (AH) is not possible with conventional assays. We investigated the use of a highly multiplexable serological assay VirScan, a Phage ImmunoPrecipitation Sequencing (PhIP-Seq) library derived from the sequences of more than 200 viruses to determine the antibody composition of AH in patients with uveitis.
Prospective, case control study.
We analyzed the paired AH and plasma samples of 11 immunocompetent patients with active polymerase chain reaction-positive cytomegalovirus (CMV) anterior uveitis and the AH of 34 control patients undergoing cataract surgery with no known uveitis in an institutional practice. The samples were tested using VirScan PhIP-Seq, and the entire pan-viral antibody repertoire was determined using peptide tile ranking by normalized counts to identify significant antibodies enrichment against all viruses with human tropism.
Significant enrichment of antibodies to Herpesviridae, Picornavirdae, and Paramyxoviridae was detectable in 20 µL of AH samples from patients with CMV uveitis and controls. Patients with CMV uveitis had relative enrichment of anti-CMV antibodies in AH compared with their plasma. Epitope-level mapping identified significant enrichment of antibodies against CMV tegument protein pp150 (P = 1.5e-06) and envelope glycoprotein B (P = .0045) in the AH compared with controls.
Our proof-of-concept study not only sheds light on the antibody repertoire of AH but also expands the utility of PhIP-Seq to future studies to detect antibodies in AH in the study of inflammatory eye diseases.
在葡萄膜炎的治疗中，明确感染病因非常重要。需要进行眼内液检测，但由于取样量有限，多重检测面临挑战。传统检测方法无法确定房水的抗体谱。我们研究了一种高度可多重检测的血清学检测方法 VirScan，该方法基于 200 多种病毒的序列构建了噬菌体免疫沉淀测序（PhIP-Seq）文库，用于确定葡萄膜炎患者房水中的抗体组成。
前瞻性病例对照研究。
我们分析了 11 例免疫功能正常的活动性聚合酶链反应阳性巨细胞病毒（CMV）前葡萄膜炎患者的配对房水和血浆样本，以及在机构实践中接受白内障手术且无已知葡萄膜炎的 34 例对照患者的房水样本。使用 VirScan PhIP-Seq 进行检测，通过归一化计数的肽图排序来确定整个泛病毒抗体谱，以识别针对所有具有人类嗜性的病毒的显著抗体富集。
在 CMV 葡萄膜炎患者和对照患者的 20 µL 房水样本中可检测到疱疹病毒科、小核糖核酸病毒科和副粘病毒科抗体的显著富集。与血浆相比，CMV 葡萄膜炎患者的房水中存在相对丰富的抗 CMV 抗体。表位水平图谱分析确定，与对照组相比，CMV 衣壳蛋白 pp150（P = 1.5e-06）和包膜糖蛋白 B（P =.0045）的抗体在房水中有显著富集。
本概念验证研究不仅阐明了房水的抗体谱，还扩展了 PhIP-Seq 在未来研究中的应用，以在炎症性眼病的研究中检测房水中的抗体。
