Role of macrophages in control of ocular HSV

 描述(由申请人提供)：巨噬细胞渗入在免疫防御系统中扮演着各种关键角色，包括在先天或自然免疫中的中心角色。通过使用抗F4/80和CD11b抗体的免疫染色，我们发现在未感染的小鼠的角膜中不能检测到常驻的角膜巨噬细胞。相比之下，在感染了单纯疱疹病毒1型(HSV-1)的小鼠之后，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中心作用。与我们以前的研究一致，我们的初步研究还表明：1.小鼠注射巨噬细胞集落刺激因子(M-CSF)DNA可减少眼部病毒复制，并减少眼部感染小鼠三叉神经节(TG)潜伏期的建立。2.与M-CSF注射相似，注射IL-12p35 DNA也可减少潜伏感染小鼠眼部病毒复制和TGS潜伏期的建立。因此，由于巨噬细胞是HSV-1眼部感染小鼠眼内最丰富的浸润细胞，也是眼部感染后出现最快的(最早4小时)，我们将使用巨噬细胞刺激因子(即M-CSF和IL-12p35)来减少病毒在眼内的复制和眼部感染小鼠TGS中潜伏期的建立。我们建议：目的：验证一种假设，即联合使用IL-12p35和M-CSF DNA作为分子佐剂刺激巨噬细胞将增强眼内TH1/Tc1应答，并导致病毒在原发眼部HSV-1感染期间更快地从眼睛和三叉神经节(TG)清除。这将导致潜伏感染小鼠TG潜伏期和重新激活的建立显著减少。我们的初步研究表明，与对照组或单独使用IL-12p35或M-CSF免疫的小鼠相比，眼部感染后眼睛内病毒复制的显著减少表明，注射IL-12p35和M-CSF DNA可以提高免疫效果。这种改善的疗效与局部眼部反应(即IL-2、干扰素-α、干扰素-？ 干扰素-γ)。我们提议检验这一假设：(1)IL-12p35和M-CSFDNA联合使用可减少病毒在眼睛中的初次复制，并缩短感染小鼠TGS中的潜伏期；(2)这些减少与除干扰素-α和干扰素外，眼部和感染小鼠的TGS中产生强烈而快速的Th1/Tc1反应有关；(3)与单独使用5GP治疗的小鼠相比，IL-12p35+M-CSF+5GP DNA联合用药将进一步缩短免疫小鼠的潜伏期。总体而言，我们希望确定IL-12p35和M-CSF的组合能够增强巨噬细胞的细胞因子分泌和抗原提呈功能，而不是其吞噬功能。

 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-α, 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-α 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.