ANALYSIS OF T CELL RESPONSES IN HUMAN LEISHMANIASIS

人类利什曼病 T 细胞反应分析

• 批准号: 6160624
• 负责人: D SACKS
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6160624
• 关键词: Macaca mulatta; Psychodidae; T lymphocyte; cellular immunity; enzyme linked immunosorbent assay; flow cytometry; gastrointestinal disorder; host organism interaction; human tissue; interferon gamma; interleukin 12; laboratory mouse; leishmaniasis; macrophage; microorganism immunology; nonhuman therapy evaluation; protozoal vaccine; tissue /cell culture; tumor necrosis factor alpha; vaccine development

Intracellular staining for cytokines and parasites, combined with two-color FACS analysis, were used to examine the frequencies of IL-12 and TNF-alpha producing macrophages in response to Leishmania infection and/or activation with IFN-gamma/LPS. Regardless of whether bone marrow-derived or inflammatory, macrophages from genetically resistant or susceptible mouse strains were used for infection, and regardless of whether the cells were infected in vitro or in vivo using metacyclic promastigotes or amastigotes, the effect of the parasite on cytokine responses by their host macrophages was identical: 1) neither IL-12 nor TNF-alpha were produced in response to infection itself; 2) virtually every infected cell lost its ability to produce IL-12 in response to IFN-gamma/LPS; and 3) the TNF-alpha response of infected cells was unimpaired. These data, generated using inflammatory macrophages, and low level, in vivo infection, reinforce the physiologic relevance of prior observations regarding the selective and potent inhibitory effect that Leishmania parasites have on IL-12 induction pathways in host macrophages. We suggest that selective impairment of IL-12 induction underlies the delayed onset of cell-mediated control mechanisms that is typical of even self-limiting forms of leishmanial disease. Studies have been initiated to evaluate the toxicity, immunogenicity and efficacy of a killed Leishmania vaccine made by B!obras in Brazil plus rhuIL-12 in monkeys (Macaca mulatta). Monkeys receiving two intradermal inoculations of aluym absorbed killed vaccine plus IL-12 were completely protected against homologous challenge with L. amazonensis. The protection was correlated with high levels of IFN-gamma in serum and in culture supernatants of PBLs stimulated with antigen in vitro.

细胞因子和寄生虫的细胞内染色，结合 双色FACS分析，用于检查IL-12的频率 和 TNF-α 产生的巨噬细胞响应利什曼原虫感染 和/或用 IFN-γ/LPS 激活。 不管有没有骨 骨髓来源的或炎症性的，巨噬细胞来自遗传抗性的 或易感小鼠品系用于感染，并且无论 细胞是否在体外或体内使用元环感染 前鞭毛体或无鞭毛体，寄生虫对细胞因子的影响 宿主巨噬细胞的反应是相同的：1) IL-12 都没有 TNF-α 也不是针对感染本身而产生的； 2） 事实上，每个受感染的细胞都失去了产生 IL-12 的能力 对 IFN-γ/LPS 的反应； 3) 感染者的TNF-α反应 细胞未受损。 这些数据是使用炎症生成的 巨噬细胞和低水平的体内感染，增强了生理学 先前关于选择性和有效的观察的相关性 利什曼原虫对 IL-12 诱导的抑制作用 宿主巨噬细胞中的途径。 我们建议选择性损害 IL-12 诱导是细胞介导控制延迟发生的基础 甚至是利什曼原虫自限性形式的典型机制 疾病。 已启动研究来评估毒性、免疫原性和 巴西 B!obras 生产的灭活利什曼原虫疫苗的功效 plus 猴子 (Macaca mulatta) 中的 rhuIL-12。 猴子收到两个 皮内接种铝吸收灭活疫苗加 IL-12 完全免受 L 的同源攻击。 亚马逊。 这种保护与高水平的 血清和 PBL 培养上清液中的 IFN-γ 刺激 体外抗原。