【作者】 郝美艳；

【导师】 王纯洁；

【作者基本信息】 内蒙古农业大学 ， 基础兽医学， 2006， 硕士

【摘要】 为了揭示鸡包涵体肝炎(Inclus Body Hepatitis,IBH)过程中肝脏内的分子免疫机理,用禽腺病毒(FAV)Ⅷ型口服接种感染1日龄SPF雏鸡,攻毒后不同日龄雏鸡病变以急性坏死性肝炎为主。取不同日龄的肝脏制作冰冻切片(或于福尔马林溶液中固定,进行石蜡包埋、石蜡切片),应用免疫组化技术检测肝脏中K1+单核巨噬细胞、CD3+、CD4+和CD8+T淋巴细胞、Bu-1a+B淋巴细胞的变化规律;采用原位杂交技术,检测肝脏枯否氏细胞(kupffer cell)、T、B淋巴细胞中的IL-2、IL-6、IFN-γ和MIP-1β的表达规律。原位杂交显示:着染部位主要出现在坏死灶或血管周围的成团或散在的淋巴细胞的胞浆内,少数出现在枯否氏细胞的胞浆内。IL-2、IL-6、IFN-γ和MIP-1β在3 dpi内表达不明显,从5 dpi表达量开始增加,9 dpi、12 dpi基本达到高峰,以后逐渐下降。免疫组化显示:K1+单核巨噬细胞3 dpi开始出现,7～9 dpi数量增多并达到高峰,12 dpi有所下降,15 dpi又增多,之后数量逐渐减少。CD3+、CD4+和CD8+T淋巴细胞3～7 dpi表达量普遍很少,之后逐渐增加, 12 dpi达到高峰,以后表达量又逐渐降低,30 dpi基本消失。Bu-1a+B淋巴细胞3～5 dpi少量表达,之后表达量持续升高,15 dpi达到高峰,以后表达量逐渐下降,30 dpi仍有少量表达。说明:鸡包涵体肝炎过程中肝脏内IL-2、IL-6、IFN-γ和MIP-1β的mRNA大量上调,可能与单核巨噬细胞和T、B淋巴细胞的增多有关;而且IL-2、IL-6、IFN-γ和MIP-1β的分泌与抗病毒感染、病症恢复及反复感染也有关。更多还原

【Abstract】 1-day-old SPF chickens were inoculated with the FAV-Ⅷ(fowl adenovirus-Ⅷ), and then scarified by collecting blood from heart in different days post inoculation (dpi). Then collected livers at different dpi, made frozen section and fixed formalin then made paraffin embedded section, and study dynamic changes of K1+ Mononuclear macrophages, CD3+、CD4+and CD8+T lymphocytes, Bu-1a+B lymphocytes by immunohistochemistry(IHC) technology ; and study dynamic changes of mRNA expressing of IL-2、IL-6、IFN-γand MIP-1βin kupffer cells、T、B lymphocytes by in situ hybridization (ISHH) technology. ISHH showed that many positive particles were seen in plasma of many T cells surrounded the blood vessels during 5 dpi～30 dpi and also a little positive particles were seen in plasma of kupffer cell during these time. They demonstrated that mRNA of IL-2、IL-6、IFN-γand MIP-1βhave less positive signal for 1～3 dpi, Expression amount increasing in 5 dpi, up to top during 9 dpi～12dpi, then fall-off. IHC showed that many positive particles were seen in K1+ mononuclear macrophages during 3 dpi and many positive particles were seen during 7 dpi～9 dpi, degrade 12dpi, increasing 15 dpi, then degrade. CD3+、CD4+and CD8+T lymphocytes e were seen a little during 3～7dpi , up to top in 12dpi, then degrade, disappeared in 30 dpi. A little positive particle were seen as Bu-1a+B lymphocytes, then increased continually, up to top at 15dpi, then degrade, a little positive particle in30dpi also. The results demonstrated that mRNA of IL-2、IL-6、IFN-γand MIP-1βwas up regulated obviously in the process of avian inclusion body hepatitis. It may be not only related with the increasing of T, B cells and kupffer cell, but also related with the IL-2、IL-6、IFN-γand MIP-1βas an immunopotentiator and chemokine in anti-virus and recovering.更多还原