Role of macrophages in control of ocular HSV

 描述（由申请人提供）：巨噬细胞浸润在免疫防御系统中发挥多种关键作用，包括在先天性或天然免疫中的中心作用。我们通过使用抗F4/80和CD 11b抗体的免疫染色显示，在未感染小鼠的角膜中检测不到常驻角膜巨噬细胞。相比之下，用单纯疱疹病毒1型（HSV-1）感染小鼠后，巨噬细胞似乎是眼睛最主要的浸润物。因此，巨噬细胞可能在急性和慢性HSV-1感染的恶化和控制中发挥核心作用。与我们以前的研究一致，我们的初步研究还表明：1。给小鼠注射巨噬细胞集落刺激因子（M-CSF）DNA可减少眼部感染小鼠的眼部病毒复制和三叉神经节（TG）潜伏期的建立。2.与M-CSF注射类似，用IL-12 p35 DNA注射小鼠也减少了眼睛中的病毒复制和潜伏感染小鼠TG中潜伏期的建立。 因此，由于巨噬细胞是HSV-1眼部感染小鼠眼中最丰富的浸润物，并且在眼部感染后在眼中出现最快（早在4小时），我们将使用巨噬细胞刺激因子（即，M-CSF和IL-12 p35）以减少眼部感染小鼠的眼部中的病毒复制和TG中潜伏期的建立。我们建议：目的：检验以下假设：IL-12 p35和M-CSF DNA作为分子佐剂组合刺激巨噬细胞将增强眼部TH 1/TC 1反应，并导致在原发性眼部HSV-1感染期间更快地清除眼部和三叉神经节（TG）中的病毒。这将导致潜伏感染小鼠的TG中潜伏期和再激活的建立显著减少。我们的初步研究表明，与对照小鼠或单独用IL-12 p35或M-CSF免疫的小鼠相比，通过眼部感染后眼睛中病毒复制的更大减少来判断，IL-12 p35和M-CSF DNA的施用改善了免疫效力。这种改善的功效与局部眼部反应的更高活性相关（即，IL-2、IFN-α、IFN-γ、IFN- γ）。我们提出检验以下假设：（1）IL-12 p35和M-CSF DNA的混合物将导致眼睛中的初级病毒复制减少以及感染小鼠的TG中的潜伏期减少;（2）这些减少与眼睛和感染小鼠的TG中除了IFN-α和IFN-γ之外的强烈和快速的TH 1/TC 1应答的产生有关;（3）IL-12 p35 + M-CSF + 5gP DNA联合免疫小鼠与单独5gP处理的小鼠相比，可进一步缩短潜伏期。总之，我们期望确定IL-12 p35和M-CSF的组合增强巨噬细胞的细胞因子分泌和抗原呈递功能，而不是它们的吞噬功能。

 DESCRIPTION (provided by applicant): Macrophage infiltrates play a variety of key roles in the immune defense system, including a central role in innate or natural immunity. We have shown by immunostaining using anti-F4/80 and CD11b antibodies that resident corneal macrophages are not detectable in cornea of uninfected mice. In contrast, following infection of mice with herpes simplex virus type-1 (HSV-1), macrophages appear to be the most dominant infiltrate of the eye. Thus, macrophages may play a central role in both exacerbation and control of acute and chronic HSV-1 infection. In line with our previous studies, our Preliminary studies also suggest that: 1. Injection of mice with macrophage-colony stimulating factor (M-CSF) DNA reduced virus replication in the eye and establishment of latency in trigeminal ganglia (TG) of ocularly infected mice. 2. Similar to M-CSF injection, injection of mice with IL-12p35 DNA also reduced virus replication in the eye and establishment of latency in TGs of latently infected mice. Thus, since macrophages are the most abundant infiltrates in the eye of HSV-1 ocularly infected mice and also appear the fastest in the eye (as early as 4 hours) after ocular infection, we will use macrophage stimulatory factors (i.e., M-CSF and IL-12p35) to reduce virus replication in the eye and establishment of latency in the TGs of ocularly infected mice. We propose to: Aim: Test the hypothesis that macrophage stimulation by a combination of IL-12p35 and M-CSF DNA as molecular adjuvants will enhance TH1/TC1 responses in the eye and lead to a more rapid clearance of virus from the eye and trigeminal ganglia (TG) during primary ocular HSV-1 infection. This will, lead to a significant reduction in establishment of latency and reactivation in TG of latently infected mice. Our preliminary studies suggest that administration of IL-12p35 and M-CSF DNA improves immunization efficacy as judged by a greater reduction in virus replication in the eye after ocular infection compare with control mice, or mice that are immunized with IL-12p35 or M-CSF alone. This improved efficacy correlated with higher activities of local ocular responses (i.e., IL-2, IFN-a, IFN-ß, IFN- γ). We propose to test the hypothesis that; (1) a cocktail of IL-12p35 and M-CSF DNA will result in reduction in primary virus replication in the eyes as well as reduction of latency in TGs of infected mice; (2) these reductions are associated with generation of strong and rapid TH1/TC1 responses in addition to IFN-a and IFN-ß in the eye and TGs of infected mice; and (3) Co-administration of IL-12p35 + M-CSF + 5gP DNA will further reduce latency in immunized mice compared with mice that treated with 5gP alone. Overall, we expect to determine that a combination of IL-12p35 and M-CSF enhances the cytokine secretion and antigen presentation functions of macrophages rather than their phagocytosis function.