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Modulation of Phenotype and Function of Monocyte-derived Macrophages Following Infection with Classical Swine Fever Virus
|關鍵字:||classical swine fever virus;豬瘟病毒;monocyte-derived macrophages;phenotype;antigen presenting cells;persistent infection;單核球衍生之巨噬細胞;表現型;抗原呈現細胞;持續性感染||出版社:||獸醫病理學研究所||摘要:||
豬瘟 (Classical swine fever, CSF) 為豬之急性病毒性疾病，造成感染豬隻免疫系統的失調，而呈現持續性感染現象。CSF virus (CSFV) 為monocytotropic virus，雖也可感染各種淋巴球次族群，但對豬肺泡巨噬細胞 (pulmonary alveolar macrophages, PAMs) 及血液中的單核球 (monocytes) 具高親和性，可於PAMs中大量複製，且不會造成PAMs的死亡。本實驗的目的為探討CSFV對活體外培養的單核球衍生之巨噬細胞 (monocyte derived macrophages, MDMs) 感染情形、表現型及功能影響。自豬隻分離出周邊血液單核細胞 (peripheral blood mononuclear cell, PBMC)，以磁性分離系統 (MACS® magnetic cell separation system) 或貼附的方法分離出單核球，經過四天培養成為MDMs後，再以0.5 MOI CSFV (病毒珠：ALD/ S59/ 94.4) 感染三天後，以anti-E2 monoclonal Ab間接螢光染色偵測CSFV於MDMs的增殖情形，及分析細胞表現型包括swine leukocyte antigen I (SLA I) 和 SLA II、吞噬功能及對不活化之PRV的抗原呈現功能。結果顯示經過四天的培養後，懸浮培養的細胞的大小及顆粒度均低、SLA分子表現低，且失去巨噬細胞的SWC3分子表現，不適合作為後續實驗分析SLA分子表現所用。貼附培養的細胞則於細胞型態、大小及表現型均較接近PAMs，顯示貼附的環境可提供MDMs較好的分化，為活體外培養MDMs的必要條件。在病毒增殖方面可發現CSFV在MDMs之增殖受到抑制，且病毒感染可降低細胞的死亡率，另一方面CSFV感染可使SLA I及SLA II分子表現下降，呈現顯著差異 (p<0.05)。在功能分析方面，CSFV感染對吞噬功能的影響無顯著差異 (p>0.05)；對抗原呈現功能也無顯著差異。整體結果顯示豬瘟病毒在MDMs中的表現受抑制，感染後可明顯降低細胞的死亡率，及降低SLA分子的表現，此些現象與豬瘟病毒之持續感染有關。
Classical swine fever (CSF) is an acute viral disease of swine. It causes severe impairment of immune modulation and results in persistent infection. CSF virus (CSFV) is a monocytotropic virus, whereas it can infect all the subpopulation of lymphocytes. CSFV can highly replicate in PAMs and not promote cell to death that suggest monocytic cells may play an important role in the persistent infection of CSFV. The aim of the study is to investigate the effect of CSFV infection on monocyte-derived macrophages (MDMs). MDMs generated from monocytes of PBMC were cultured by suspension or adherence for 4 days. The cells were infected with 0.5 MOI different strains of CSFV (ALD, s59, 94.4) for 3 days, and the expression of CSFV E2 antigen in MDMs were then detected. Further analysis the effect on expression of SLA I and SLA II molecules, phagocytosis and antigen presenting function of MDMs after CSFV infection was undertaken. The results showed that the suspensively cultured cells had smaller cell size, lower cytoplasmic granules and SLA expression compared with PAMs. The loss of the macrophage marker-SWC3 expression was also noted. The adherently cultured cells had similar cell morphology and phenotype to PAMs. It reveals the adherent condition is essential for MDMs culture. Therefore, the adherent culture system was used in following assay. Compared with PAMs, the replication of CSFV in MDMs were limited. CSFV infections delayed MDMs toward to death and down regulated the expression of SLA I and SLA II (p<0.05). However there were no significant difference in the function assays of phagocytosis and antigen presentation. Conclusively, CSFV replicated limitedly in MDMs, down regulated SLA molecule and extended the survival of MDMs, suggesting MDMs may play an important role in the persistent infection of CSFV.
|Appears in Collections:||獸醫病理生物學所|
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