TY - JOUR
T1 - Immunopathological characterization of porcine circovirus type 2 infection-associated follicular changes in inguinal lymph nodes using high-throughput tissue microarray
AU - Lin, Chun Ming
AU - Jeng, Chian Ren
AU - Hsiao, Shih Hsuan
AU - Liu, Jen Pei
AU - Chang, Chih Cheng
AU - Chiou, Ming Tang
AU - Tsai, Yi Chieh
AU - Chia, Mi Yuan
AU - Pang, Victor Fei
N1 - Funding Information:
We would like to thank Dr. M.S. Lilburn, Department of Animal Sciences, the Ohio State University and Dr. S.T. Ding, Department of Animal Science and Technology, National Taiwan University for their reading and suggestions on manuscript preparation. This research was supported in part by grant NSC 98-2313-B-002-025-MY3 from the National Science Council of the Republic of China.
PY - 2011/4/21
Y1 - 2011/4/21
N2 - 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.
AB - 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.
KW - Immunopathology
KW - Lymphoid follicle
KW - Porcine circovirus type 2 (PCV2)
KW - Postweaning multisystemic wasting syndrome (PMWS)
KW - Tissue microarray (TMA)
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U2 - 10.1016/j.vetmic.2010.10.018
DO - 10.1016/j.vetmic.2010.10.018
M3 - Article
C2 - 21126833
AN - SCOPUS:79952107078
SN - 0378-1135
VL - 149
SP - 72
EP - 84
JO - Veterinary Microbiology
JF - Veterinary Microbiology
IS - 1-2
ER -