The role of gK in HSV-1 induced corneal scarring

gK 在 HSV-1 诱导的角膜疤痕中的作用

• 批准号: 7459964
• 负责人: HOMAYON GHIASI
• 金额: $ 36.56万
• 依托单位: CEDARS-SINAI MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-09 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7459964
• 关键词: Acute; Address; Adoptive Transfer; Affect; Antibodies; Antigen Receptors; Antigens; Apoptosis; Appendix; Area; Attenuated; B-Lymphocytes; Baculoviruses; Binding; Blepharitis; Blindness; CD8-Positive T-Lymphocytes; CD8B1 gene; Cells; Clinical; Conjunctivitis; Cornea; Corneal Diseases; Data; Dendritic Keratitis; Deposition; Developed Countries; Developing Countries; Disease; Edema; Elevation; Eye; Eye Infections; Eye diseases; Genes; Glycoproteins; Herpesvirus 1; Human; IL2RA gene; Immune; Immune response; Immune system; Immunization; In Vitro; Infection; Infectious Agent; Keratitis; Keratoplasty; Kinetics; Lead; Life; Link; MHC Class I Genes; Maps; Masks; Mouse Strains; Mus; Numbers; Outcome; Peptides; Personal Satisfaction; Pharmaceutical Preparations; Phenotype; Play; Population; Prevention; Process; Protein Overexpression; Proteins; Range; Recombinants; Recurrence; Role; Role playing therapy; Severities; Simplexvirus; Site; Specificity; T-Lymphocyte; TNFRSF5 gene; Testing; Time; Vaccinated; Vaccines; Viral; Viral Eye Infections; Viral Genes; Viral Proteins; Virus; Visit; base; corneal scar; cytokine; day; interest; protein function; response; synthetic peptide

DESCRIPTION (provided by applicant): It is well established that HSV-1-induced corneal scarring (CS), and thus HSV-1-induced corneal blindness, are the result of immune responses triggered by the virus. The exact identity of the immune responses, including the fine specificity of the potentially harmful T-cell effectors expressing classic TCR12 antigen receptors that lead to CS remains an area of intense controversy. The vast majority of non-ocular HSV infections are either entirely asymptomatic or so mildly symptomatic as to go almost completely unrecognized. In contrast, symptomatic eye disease after HSV-1 reactivation is a major cause of corneal disease and blindness. The damaging effects of pre-existing immune responses that are manifested during recurrent infections is one rational explanation for the clinical observation that CS in humans is much more likely to occur following recurrent HSV infections rather than on primary infections. Our previous (Appendix 1) and preliminary results demonstrate an absence of CD8+ T cells in the cornea of mice infected with HSV-1. This absence of CD8+ T cells in the eyes of infected mice was observed irrespective of the strain of virus used for infection or the strain of mouse infected. Notably, however, we have found that infection of mice that have been immunized with one of the HSV-1 glycoproteins, gK, both exacerbates CS and results in the presence of CD8+CD25+ cells in the cornea. The exacerbation of CS in the immunized mice is associated with the presence of gK antibody and that the CD8+ cells are detectable in the corneas of mice immunized with gK, but not with live virus vaccine or any of the other known HSV-1 glycoproteins. This exacerbation of disease by an existing immune response in mice is of particular interest as it would appear to mimic the clinical disease process. Based on the preliminary data we have now generated, we hypothesize that, in unimmunized mice, viral gK acts to block induction of CD8+CD25+ T cells in the cornea of infected mice. In immunized mice, the gK-specific antibody binds the viral gK and masks the sites required for its inhibitory effects on the CD8+ T cells. Thus, "antigenic masking", independent of MHC class I or any other viral gene(s) in gK-immunized mice, leads to the stimulation of CD8+CD25+ T cells in the cornea of infected mice. The subsequent increase in the population of CD8+CD25+ T cells in the cornea leads to exacerbation of CS. We propose to test this hypothesis through the following Specific Aims: Aim 1: Confirm the hypothesis that anti-gK antibody play a major role in the induction of CD8+CD25+ T cells in the cornea of gK-immunized mice. Aim 2: Confirm the hypothesis that, in gK-immunized mice, the presence of CD8+CD25+ T cells in the cornea is responsible for exacerbation of CS on HSV-1 infection. Aim 3: Map the gK region that is involved in stimulation of CD8+CD25+ T cells and exacerbation of CS.HSV-1 infections are among the most frequent serious viral eye infections in the U.S. and are a major cause of viral-induced blindness. HSV-1-induced CS, also broadly referred to as herpes stromal keratitis (HSK), can lead to blindness; thus, HSV-1 is the leading cause of corneal blindness due to an infectious agent in developed countries. In addition to necrotizing stromal keratitis, ocular infection with HSV-1 can cause eye disease ranging in severity from blepharitis, conjunctivitis, and dendritic keratitis, to disciform stromal edema. In the U.S., approximately 500,000 people suffer recurrent ocular HSV episodes annually, requiring doctor visits, medication, and, in severe cases, corneal transplants. We have shown for the first time that one of more than 80 genes of HSV-1 exacerbates eye disease in ocularly infected mice. We now plan to determine the mechanism of gK induced corneal scarring mice. These studies should reveal the type or combination of types of immune response that induces eye disease.

描述（由申请人提供）：已经确定hsv -1诱导的角膜瘢痕（CS）以及由此引起的hsv -1诱导的角膜失明是由该病毒引发的免疫反应的结果。免疫反应的确切身份，包括表达经典TCR12抗原受体的潜在有害t细胞效应物的精细特异性，导致CS仍然是一个激烈争议的领域。绝大多数非眼部HSV感染要么完全无症状，要么症状非常轻微，几乎完全无法识别。相反，HSV-1再激活后的症状性眼病是角膜疾病和失明的主要原因。在复发性感染期间表现出的预先存在的免疫反应的破坏性影响是临床观察的一个合理解释，即人类CS更可能发生在复发性HSV感染之后，而不是发生在原发性感染之后。我们之前的（附录1）和初步结果表明，感染HSV-1的小鼠角膜中缺乏CD8+ T细胞。无论用于感染的病毒株或感染的小鼠株是什么，在感染小鼠的眼睛中都观察到CD8+ T细胞的缺失。然而，值得注意的是，我们发现用HSV-1糖蛋白之一gK免疫的小鼠感染，既加剧了CS，又导致角膜中CD8+CD25+细胞的存在。免疫小鼠CS的恶化与gK抗体的存在有关，并且在gK免疫小鼠的角膜中检测到CD8+细胞，但在活病毒疫苗或任何其他已知的HSV-1糖蛋白中检测不到。这种由小鼠现有免疫反应引起的疾病恶化是特别有趣的，因为它似乎模仿了临床疾病过程。根据我们现在获得的初步数据，我们假设，在未免疫的小鼠中，病毒gK可以阻断感染小鼠角膜中CD8+CD25+ T细胞的诱导。在免疫小鼠中，gK特异性抗体结合病毒gK并掩盖其对CD8+ T细胞抑制作用所需的位点。因此，在gk免疫小鼠中，独立于MHC I类或任何其他病毒基因的“抗原掩蔽”导致感染小鼠角膜中CD8+CD25+ T细胞的刺激。随后角膜中CD8+CD25+ T细胞群的增加导致CS的恶化。我们提出通过以下具体目的来验证这一假设：目的1：证实抗gk抗体在gk免疫小鼠角膜诱导CD8+CD25+ T细胞中起主要作用的假设。目的2：证实在gk免疫小鼠中，角膜中CD8+CD25+ T细胞的存在是导致HSV-1感染后CS恶化的原因。目的3：绘制参与CD8+CD25+ T细胞刺激和CS加重的gK区域。1型单纯疱疹病毒感染是美国最常见的严重病毒性眼部感染之一，也是病毒性失明的主要原因。hsv -1诱导的CS，也被广泛地称为疱疹间质角膜炎（HSK），可导致失明；因此，在发达国家，单纯疱疹病毒-1是一种传染性病原体导致角膜失明的主要原因。除了坏死性间质角膜炎外，眼部感染HSV-1还可引起严重程度不等的眼部疾病，从睑缘炎、结膜炎、树突状角膜炎到盘状间质水肿。在美国，每年大约有50万人患有反复发作的眼部HSV，需要看医生，服用药物，在严重的情况下，需要角膜移植。我们首次证明了80多个HSV-1基因中的一个会加重眼部感染小鼠的眼病。我们现在计划确定gK诱导小鼠角膜瘢痕形成的机制。这些研究应该揭示引起眼病的免疫反应的类型或组合。