Endothelial cell injury by cytolytic lymphocytes

溶细胞淋巴细胞损伤内皮细胞

• 批准号: 7050548
• 负责人: Mitzi Nagarkatti
• 金额: $ 23.07万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7050548
• 关键词: CD44 molecule; CD95 molecule; cell cell interaction; cellular immunity; cytolysins; cytotoxic T lymphocyte; cytotoxicity; electron microscopy; enzyme linked immunosorbent assay; flow cytometry; genetically modified animals; high performance liquid chromatography; histochemistry /cytochemistry; interleukin 2; laboratory mouse; ligands; lymphokine activated killer cell; major histocompatibility complex; natural killer cells; polymerase chain reaction; pore forming protein; tissue /cell culture; vascular endothelium

DESCRIPTION (provided by applicant): Endothelial cell (EC) injury has been widely documented in certain infections, autoimmunity, allograft rejection, graft versus host disease and following immunotherapy with cytokines or immunotoxins. It is becoming increasingly clear that ECs are primary targets of immunologic attack, although the precise mechanism remains unclear. We have used interleukin-2 (IL-2)-induced vascular leak syndrome (VLS) as a model to study EC injury. IL-2 is the only FDA approved systemic therapy for treating metastatic renal cell carcinoma. It is also being used to treat metastatic melanomas, immunodeficiencies and viral infections including AIDS. Despite much success, the efficacy is limited by severe, life-threatening toxicity resulting from EC injury leading to VLS. Using CD44 knockout (KO), perforin KO and FasL mutant mice, we have demonstrated that IL-2 activates cytotoxic lymphocytes and such cells use CD44 to kill EC by triggering perforin and FasL. More recently, we have demonstrated that CD44v7 exon KO mice also exhibit decreased VLS and EC killing. Based on these studies, we will test the hypothesis that IL-2 treatment up regulates CD44 variant isoforms on cytolytic lymphocytes of which v7 isoform plays a key role in EC injury and induction of VLS. The specific aims are 1) To test the susceptibility of EC isolated from various organs to cytotoxicity mediated by IL-2 activated T/NK/NKT cells from WT, pefforin KO, Fas and FasL mutant mice. Using NK cell-deficient (NKCD-Tg) mice, the role of NK cells in IL-2 induced VLS will also be investigated. 2) To examine whether down regulation of killer cell inhibitory receptor (KIR)-like molecules and/or up regulation of stimulatory receptors on NK cells is responsible for CD44-mediated EC injury. Specifically, the expression of KiR-like molecules, including Ly49 and CD94/NKG2 present on NK cells, and the classical and non-classical MHC ligands on Ecs, will be studied. 3) Identification of the CD44 isoforms involved in EC injury caused by cytotoxic lymphocytes. 4) To address the mechanism by which CD44v7KO mice exhibit increased resistance to VLS. 5) Use of mimetics of CD44 or its ligand including mAbs against CD44s and CD44v isoforms, Pep-l(an inhibitor of HA), CD44Rg and CD44MutRg fusion proteins to prevent EC injury and vascular leak. Together, the current study should provide novel insights into the mechanism of immune cell-mediated EC injury and development of strategies to prevent vascular leak.

描述（由申请人提供）：内皮细胞（EC）损伤在某些感染、自身免疫、同种异体移植排斥、移植物抗宿主病和细胞因子或免疫毒素免疫治疗后被广泛记录。越来越清楚的是，ECs是免疫攻击的主要目标，尽管其确切机制尚不清楚。我们以白细胞介素-2 （IL-2）诱导的血管渗漏综合征（VLS）为模型研究EC损伤。IL-2是FDA批准的唯一治疗转移性肾细胞癌的全身疗法。它也被用于治疗转移性黑色素瘤、免疫缺陷和包括艾滋病在内的病毒感染。尽管取得了很大的成功，但由于EC损伤导致VLS的严重，危及生命的毒性，其疗效受到限制。使用CD44敲除（KO）、穿孔素KO和FasL突变小鼠，我们已经证明IL-2激活细胞毒性淋巴细胞，这些细胞使用CD44通过触发穿孔素和FasL来杀死EC。最近，我们已经证明CD44v7外显子KO小鼠也表现出VLS和EC杀伤降低。基于这些研究，我们将验证IL-2处理上调细胞溶解淋巴细胞上CD44变异亚型的假设，其中v7亚型在EC损伤和VLS诱导中起关键作用。1)检测不同器官EC对WT、peforin KO、Fas和FasL突变小鼠IL-2活化T/NK/NKT细胞介导的细胞毒性的敏感性。使用NK细胞缺陷（NKCD-Tg）小鼠，NK细胞在IL-2诱导的VLS中的作用也将被研究。2)探讨NK细胞上杀伤细胞抑制受体（KIR）样分子的下调和/或刺激受体的上调是否与cd44介导的EC损伤有关。具体来说，我们将研究NK细胞上的kir样分子，包括Ly49和CD94/NKG2的表达，以及Ecs上的经典和非经典MHC配体。3)细胞毒性淋巴细胞致EC损伤相关CD44亚型的鉴定。4)探讨CD44v7KO小鼠对VLS抗性增强的机制。5)利用CD44或其配体的模拟物，包括针对CD44s和CD44v亚型、pep - 1 （HA抑制剂）、CD44Rg和CD44MutRg融合蛋白的单克隆抗体，预防EC损伤和血管泄漏。总之，目前的研究应该为免疫细胞介导的EC损伤机制和预防血管泄漏策略的发展提供新的见解。

项目成果

Combined deficiency in CD44 and Fas leads to exacerbation of lymphoproliferative and autoimmune disease.

CD44 和 Fas 的联合缺乏会导致淋巴增殖性疾病和自身免疫性疾病的恶化。

• DOI: 10.1093/intimm/dxg132
• 发表时间: 2003
• 期刊: International immunology
• 影响因子: 4.4
• 作者: Do,Yoonkyung;Rafi-Janajreh,AsimahQ;McKallip,RobertJ;Nagarkatti,PrakashS;Nagarkatti,Mitzi
• 通讯作者: Nagarkatti,Mitzi

Endocannabinoids and immune regulation.

• DOI: 10.1016/j.phrs.2009.03.019
• 发表时间: 2009-08
• 期刊: PHARMACOLOGICAL RESEARCH
• 影响因子: 9.3
• 作者: Pandey, Rupal;Mousawy, Khalida;Nagarkatti, Mitzi;Nagarkatti, Prakash
• 通讯作者: Nagarkatti, Prakash

Growth of FasL-bearing tumor cells in syngeneic murine host induces apoptosis and toxicity in Fas(+) organs.

携带 FasL 的肿瘤细胞在同基因小鼠宿主中的生长会诱导 Fas( ) 器官的细胞凋亡和毒性。

• 发表时间: 2000
• 期刊: Blood
• 影响因子: 20.3
• 作者: Zeytun,A;Nagarkatti,M;Nagarkatti,PS
• 通讯作者: Nagarkatti,PS

Immune modulation by chondroitin sulfate and its degraded disaccharide product in the development of an experimental model of multiple sclerosis.

• DOI: 10.1016/j.jneuroim.2010.04.002
• 发表时间: 2010-06
• 期刊: Journal of neuroimmunology
• 影响因子: 3.3
• 作者: Zhou J;Nagarkatti P;Zhong Y;Nagarkatti M
• 通讯作者: Nagarkatti M

Evaluation of Cell Proliferation and Apoptosis in Immunotoxicity Testing.

• DOI: 10.1007/978-1-4939-8549-4_14
• 发表时间: 2018
• 期刊: Methods in molecular biology
• 作者: M. Nagarkatti;S. Rieder;P. Nagarkatti