Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/12778
標題: 豬肺泡巨噬細胞之表現型和功能及假性狂犬病病毒對其之影響
PhenotyPhenotype and phagocytic activities of swine alveolar macrophages and the effects after
作者: 廖偉莉
Liao, Wei Li
關鍵字: alveolar macrophages;肺泡巨噬細胞;phenotype;phagocytosis;pseudorabies;表現型;吞噬作用;假性狂犬病
出版社: 獸醫學系
摘要: 
摘要豬呼吸道疾病為現代養豬事業的一大問題,造成嚴重經濟損失。肺泡
巨噬細胞 (pulmonary alveolar macrophages; PAM) 為下呼吸道的第一
道防線,因此,探索豬隻PAM之表現型與功能,及病原對其之影響有助於
了解豬隻為何易罹患肺炎之機制。本實驗選取健康之哺乳小豬,隔離飼養
,於不同生長階段,進行肺部灌洗,以流體細胞儀分析PAM之表現型及功
能,並評估年齡因子對PAM之影響。並將此基本資料與同年齡層之SPF小豬
、豬生殖與呼吸道綜合症病毒及沙氏桿菌感染豬之PAM相比較。結果顯示
,3週齡SPF豬SLA II表現量顯著高於各年層豬隻;而SLA I及macrophages
specific molecule等分子表現量,以4-8週齡表現最高,此後隨年齡之增
加而有下降現象。在吞噬功能方面,年齡層間並無顯著差異。但豬生殖與
呼吸道綜合症病毒感染豬之PAM吞噬能力,顯著低於同齡的豬隻。沙氏桿
菌感染豬PAM之SLA I及macrophage分子亦均顯著低於同齡的豬隻。進而以
假性狂犬病 (pseudorabies; PR) 病毒 (PRV) 為模式,探討PRV對PAM之
影響及病毒感染後對肺微環境之影響。在in vitro方面,利用SPF豬之PAM
,以PRV感染 (台灣台南株,1 MOI),並於感染後不同時段收集細胞,進
行表現型及功能分析。結果顯示,PAM對PRV具有高度之感受性,並於感染
後6小時即造成其表面分子SLA I及SLA II的減少,但對RBC之吞噬作用及
表面FcR的表現並無顯著之影響。此外,以PR中和抗體力價低於2倍以下之
生長期豬隻,經麻醉後由肺泡腔接種PRV (1×107 TCID50),並於感染後0
、7、12、18天,進行肺泡腔細胞灌洗及PAM表現型及功能分析、病毒抗原
偵測,病理檢查及分析肺淋巴細胞在感染期間可能扮演之角色。結果顯示
由肺部直接感染PRV造成豬隻短暫食慾減退,並造成輕微之間質性肺炎,
其中二頭亦併發輕度之纖維素胸膜肺炎。實驗組之PAM對Pasteurella
multocida吞噬能力有被抑制現象。在PRV感染後12天可於灌洗液中之PAM
偵測到少量病毒存在;相對地在扁桃腺、肺門淋巴結、肺中膈淋巴結等淋
巴組織,則有較高之感染率。進一步分析肺灌洗液內淋巴細胞,發現在感
染後CD4+及CD8+淋巴細胞表現較多之SLA II分子,且有較多數之CD4-CD8-
淋巴細胞被轉化為CD4+CD8+淋巴細胞,且對PAM具有毒殺作用,但無MHC限
制及無病毒特異性,推測可能主要由NK及LAK細胞媒介此毒殺作用。

SummaryRespiratory disorders are regarded as the serious problem
in modern swine industry. Pulmonary alveolar macrophages (PAM)
are the center in defending the lungs against particles and
pathogens in inspired air. Therefore, the investigation of the
phenotype and basic function of PAM and the alterations after
infections are critical to know the complexity of pulmonary
diseases. The experiment was first studied on the phenotype and
phagocytosis of PAM of pigs from different age groups by flow
cytometry and then compared to the data analyzed from SPF pigs
and pigs naturally infected with porcine reproductive and
respiratory syndrome (PRRS) virus and Salmonella choleraesuis.
The result showed that the expression of SLA I and macrophage
markers of PAM from the age of 4- and 8-week old pigs were
significantly higher than other pigs. However phagocytosis of
goat RBC and RBC rosetting activities of PAM were no significant
difference in various age group. In contrast to normal pigs, 3
week-old SPF pigs had more SLA II-DR expression on PAM (P<0.05).
SLA I and macrophage marker of PAM in S. choleraesuis infected
pigs were significantly decreased. Phagocytic activity of PAM
from PRRS virus infected pigs was significantly decreased when
compared to the pigs with the same age (P<0.05). Secondary, we
studied on the interaction of pseudorabies and PAM in vitro and
the effect on the pulmonary microenvironment by PRV infection in
vivo. In vitro study showed that PAM was susceptible for PRV
infection. SLA I and SLA II expression on PRV infected PAM were
decreased, but phagocytosis of RBC and RBC rosetting activity
were not affected. In vivo we used 15-week-old pigs and infected
them with PRV endobronchially. BAL samples were periodically
collected on day 0, 7, 12, and 18 post inoculation (DPI). Viral
antigen expression was detected by IFA and phenotype and
functional activities of PAM were assayed by flow cytometry.
Pigs inoculated with PRV lost their appetite for a few days and
then recovered. All PRV inoculated pigs developed more serious
interstitial pneumonia than controls and two of them also showed
mild chronic pleuritis. PRV antigen was more easily detected in
lymphoid tissues rather than PAM on 12 DPI. Moreover, PAM from
PRV infected pigs decreased their phagocytic activity to P.
multocida as compared to PAM function before virus infection. In
addition to PAM, phenotype and cytotoxic function of
bronchoalveolar lavage (BAL) lymphocytes were also assessed. The
result displayed that more CD4+ and CD8+ lymphocytes expressed
SLA II molecules after PRV infection and the increase of CD4+
CD8+ lymphocytes in BAL might be activated from CD4-CD8-
lymphocytes. BAL lymphocytes, which had been activated in vitro
in the presence of PRV antigen and exogenous rhIL-2 for 3 days,
showed non-MHC restricted and non-virus specific cytotoxicity to
PAM. Therefore, natural killer cells or lymphokine activated
killer cells might play more contribution in control PRV
replication in pulmonary environment. The defect of PAM function
after PRV infection might contribute to the secondary infection
in pulmonary diseases.
URI: http://hdl.handle.net/11455/12778
Appears in Collections:獸醫學系所

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