Immunopathological characterization of porcine circovirus type 2 infection-associated follicular changes in inguinal lymph nodes using high-throughput tissue microarray
Immunopathological characterization of porcine circovirus type 2 infection-associated follicular changes in inguinal lymph nodes using high-throughput tissue microarray
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DOI：
10.1016/j.vetmic.2010.10.018
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发表时间：
2011-04-21
影响因子：
3.3
通讯作者：
Pang, Victor Fei
中科院分区：
文献类型：
--
作者：
Lin, Chun-Ming;Jeng, Chian-Ren;Pang, Victor Fei
常规猪感染猪圆环病毒2型（PCV2）的免疫发病机制复杂，受多种环境因素和个体差异影响，难以在实验上完全重现。在本现场研究中，使用来自 102 头猪的福尔马林固定和石蜡包埋的浅腹股沟淋巴结 (LN) 构建了由一系列具有不同 PCV2 载量的淋巴滤泡组成的组织微阵列 (TMA)。使用 TMA，通过免疫组织化学 (IHC) 染色或原位杂交 (ISH) 测量、表征和比较多种参数，包括共感染的病毒病原体、免疫细胞亚群以及细胞凋亡/增殖活性。每个参数的信号位置和面积范围由病理学家解释，通过自动图像分析软件半量化，并进行统计分析。本文的结果表明，使用 Pearson 相关分析，PCV2 和 CD79a (p < 0.001) 之间存在显着负相关，而 PCV2 和溶菌酶 (p < 0.001) 或 TUNEL (p < 0.001) 之间存在显着正相关。猪呼吸与繁殖综合征病毒（PRRSV）和猪细小病毒抗原的量与PCV2核酸的组织负荷不相关。多元回归分析进一步预测PCV2对CD79a、溶菌酶和TUNEL产生主要影响，但PRRSV对这些参数的影响相对较小。此外，Ki67（细胞增殖活性指数）的总信号强度在不同PCV2载量的病例中没有显着变化；然而，随着PCV2核酸负载量的增加，Ki67信号的主要贡献逐渐从生发中心的B细胞转移到滤泡间区域的T细胞和巨噬细胞。在本研究中，利用TMA建立具有更广泛统计分析范围的数学模型，可以使我们在了解PCV2感染相关的LN滤泡变化的免疫发病机制方面向前迈进一步。 (C) 2010 Elsevier B.V. 保留所有权利。
The immunopathogenesis of porcine circovirus type 2 (PCV2) infection in conventional pigs is complicated by various environmental factors and individual variation and is difficult to be completely reproduced experimentally. In the present field-based study, a tissue microarray (TMA) consisting of a series of lymphoid follicles having different PCV2-loads was constructed using formalin-fixed and paraffin-embedded superficial inguinal lymph nodes (LNs) from 102 pigs. Using the TMA, a wide range of parameters, including co-infected viral pathogens, immune cell subsets, and cell apoptosis/proliferation activity by immunohistochemical (IHC) staining or in situ hybridization (ISH) were measured, characterized, and compared. The signal location and area extent of each parameter were interpreted by pathologists, semi-quantified by automated image analysis software, and analyzed statistically. The results herein demonstrated a significant negative correlation between PCV2 and CD79a (p < 0.001) and a significant positive correlation between PCV2 and lysozyme (p < 0.001) or TUNEL (p < 0.001) using Pearson correlation analysis. The amount of porcine respiratory and reproductive syndrome virus (PRRSV) and porcine parvovirus antigens did not correlate with the tissue loads of PCV2 nucleic acid. Multiple regression analysis further predicted that PCV2 contributed major effects on CD79a, lysozyme, and TUNEL but PRRSV showed relatively less effects on these parameters. In addition, the total signal intensity of Ki67 (index of cell proliferation activity) did not change significantly among cases with different PCV2 loads; however, as the loading of PCV2 nucleic acid increased, the main contribution of Ki67 signal gradually shifted from B cells in the germinal center to T cells and macrophages in the interfollicular regions. In the present study, the use of TMA to establish a mathematical model with a wider range of statistical analysis can bring us a step forward to understand the immunopathogenesis of PCV2 infection-associated follicular changes in LNs. (C) 2010 Elsevier B.V. All rights reserved.