IMMUNE FUNCTIONS OF ACCESSORY CELLS IN THE LUNG

拟议研究的长期目标是分析 肺辅助细胞免疫功能及其与免疫功能的关系 异源抗原引起的免疫反应。树枝状 细胞(DC)，最强大的辅助细胞之一，最近被认为是 从肺和上呼吸道鉴定和隔离。在这 建议，实验旨在表征表型和 Lewis大鼠上呼吸道和肺的DC功能 检查可能涉及其监管的因素。具体的 目的是通过超微结构、免疫细胞化学来确定 胎/新生大鼠肺和上呼吸道树突状细胞的个体发育 为了在体外检测这些新分化的细胞的能力 诱导抗原(鸡蛋溶菌酶，HEL)和凝集素(ConA)介导的T- 细胞增殖。细胞因子(IL-1、IL-6、GM-CSF、干扰素-α) 伽马)，以加速这些早期DC的成熟将受到考验。 树突状细胞向肺和局部淋巴结的迁移模式为 在静脉注射111进行的生物分布研究中进行检查 In标记的低密度骨髓DC前体细胞。紧随其后的是 ~H-尿苷标记DC静脉注射后在体内的分布 组织将通过免疫细胞化学放射自显影来确定 肺和上呼吸道的反应切片。使用纯化的DC 从成年大鼠的肺中分离出单抗(MAb) 开发了与大鼠DC特异反应的药物，用于鉴定和 分离这些细胞。将对单抗进行进一步筛选，以确定 识别DC表面假定黏附分子的单抗(S) 介导T细胞结合的分子，以及有丝分裂分子(S) 刺激T细胞增殖。研究将抑制物和细胞因子 在分离的肺DC上，将检查DC对T细胞的要求 集群化。细胞因子(IL-1、IL-4、IL-6、GM-CSF、肿瘤坏死因子) α和干扰素-γ)调节肺树突状细胞的辅助细胞功能， 同种异体MLR与抗原和凝集素介导的T细胞 反应，将在体外和体内进行测试。有没有可能 淋巴因子激活的杀伤细胞(LAK)可能下调DC活性 将从IL-2处理的大鼠肺分离的DC进行体外检测。 相反，IL-2或体外处理肺树突状细胞 生成的LAK细胞将用于确定IL-2或LAK细胞 直接影响DC在培养过程中的辅助细胞功能。结果来自 这些研究应该有助于更好地理解区议会在 肺的免疫防御及其调节因素 活动。

The long term objective of the proposed research is to analyze the immune function of accessory cells in the lung and to relate these to immune responses triggered as a result of foreign antigens. Dendritic cells (DC), one of the most potent accessory cells, have recently been identified and isolated from the lung and upper airways. In this proposal, experiments are designed to characterize phenotypically and functionally the DC of upper airways and lungs of Lewis rats ad to examine factors that may be involved in their regulation. The specific aims are to determine by ultrastructural, immunocytochemistry the ontogeny of DC in the lung and upper airways of fetal/newborn rats and to examine in vitro the ability of these newly differentiated cells to induce antigen (hen egg lysozyme, HEL) and lectin mediated (Con A) T- cell proliferation. The capacity of cytokines (IL-1, IL-6, GM-CSF, IFN- gamma) to accelerate the maturation of these early DC will be tested. The migration pattern of DC to the lung and local lymph nodes will be examined in a bio-distribution study using intravenously injected 111 In-labeled, low density bone marrow DC precursors. Following the intravenous injection of 3 H-uridine labeled DC, the distribution in tissue will be determined by autoradiography on immunocytochemically reacted sections of the lung and upper airways. Using purified DC isolated from adult rat lungs, monoclonal antibodies (MAb) will be developed that react specifically with rat DC for the identification and isolation of these cells. The MAb will be screened further to identify MAb that recognize, on the surface of DC, putative adhesion molecule(s) that mediate T-cell binding, as well as mitogenic molecule(s) that stimulate T-cells proliferation. Studies will inhibitors and cytokines on isolated lung DC, will examine the requirements for DC to T-cell clustering. The capacity of cytokines (IL-1, IL-4, IL-6, GM-CSF, TNF alpha and IFN-gamma) to regulate the accessory cell function of lung DC, as measured by allogeneic MLR and antigen and lectin mediated T-cell responses, will be tested in vitro and in vivo. The possibility that lymphokine activated killer (LAK) T-cells may down-regulate DC activity will be examined in vitro on lung DC isolated from IL-2 treated rats. Conversely, treatment in vitro of lung DC with either IL-2 or in vitro generated LAK cells will be used to determine whether IL-2 or LAK cells directly affect accessory cell function of DC in culture. Results from these studies should lead to a better understanding of the role of DC in the immune defense of the lung and the factors that regulate their activities.