FACTORS AFFECTING GROWTH OF BRUGIA MALAYI IN SCID MICE

这一提议将检验这样一种假设，即成功地消除 小鼠宿主的马来丝虫需要 适应性和先天免疫系统。我们假设(A)适应性 免疫活性小鼠免疫系统对L3免疫原(S)的应答 幼虫(B)上调先天免疫系统的活性， 可能是通过分泌细胞因子，如肿瘤坏死因子-α和干扰素-γ。我们 进一步假设(C)免疫系统的组件(例如 巨噬细胞)然后分泌各种化学介质，包括，但是 不限于一氧化氮，(D)对传染性疾病造成损害 探员。我们将审查； 1.正常小鼠对攻击感染的早期免疫反应：WE 将审查(A)。引流淋巴中免疫反应的动态变化 结节和脾，用流动显微荧光法，用试剂 定量测定B细胞、α-βT淋巴细胞(CD4T和CD8细胞)、 Gammadelta T淋巴细胞，以及活化标志物如 CD44和CD45Ro。(B)。T的不同Vbeta亚集的反应动力学 引流淋巴结中的CD4和CD8亚群中的淋巴细胞 脾/(C)。淋巴引流分泌细胞因子的研究进展 结节细胞，通过直接的酶联免疫吸附试验和半定量RT- 聚合酶链式反应。 2.MHC-II和MHC-II的抗丝虫免疫机制 TCRalphabeta基因敲除小鼠对挑战感染具有抵抗力。我们 将审查(A)。这些基因敲除小鼠的驱虫动力学 与正常人相比。(B)。排水过程中的细胞种群动态 在特定目的中使用上述细胞表面标记的淋巴转移 #1.(C)引流淋巴结分泌细胞因子的研究进展 细胞，通过直接的ELISA法和半定量RT-PCR法。 3.一氧化氮在宿主抵抗丝虫感染中的作用。 我们将检查(A)马来丝虫在小鼠体内的生长情况，在这种情况下， 一氧化氮合酶(INOS)基因被同源基因灭活 重组。(B)Nramp和iNOS基因座上的等位基因对 小鼠对马来丝虫的易感性。(C)。细胞因子的调控作用 NO诱导NOD/LtSz-SCID小鼠对B.B. 马来人。

This proposal will test the hypothesis that the successful elimination of Brugia malayi by the murine host requires the concerted action of the adaptive and innate immune systems. We hypothesize that (a) the adaptive immune system in immunocompetent mice responds to immunogen(s) of the L3 larvae to (b) up-regulate the activity of the innate immune system, probably by the secretion of cytokines like TNF-alpha and IFN-gamma. We further hypothesize that (c) components of the immune system (such as macrophages) then secrete a variety of chemical mediators, including, but not limited to nitric oxide, that (d) inflict damage on the infectious agent. We will examine; 1. The early immune response of normal mice to challenge infection: We will examine the (a). dynamics of the immune response in draining lymph nodes and the spleen, by flow microfluorimetry, using reagents to quantitate B cells, alpha beta T lymphocytes (CD4 T and CD8 cells), gammadelta T lymphocytes, and the expression of activation markers such as CD44 and CD45Ro. (b). kinetics of response of various Vbeta subsets of T lymphocytes, both in the CD4 and CD8 subsets ina the draining lymph node and in the spleen/ (c). evolution of cytokine secretion by draining lymph node cells, both by direct ELISA assays, as well as semi-quantitative RT- PCR. 2. The mechanism of anti-filarial immunity in MHC class II and TCRalphabeta knock-out mice that are resistant to challenge infection. We will examine the (a). kinetics of worm elimination in these knock-out mice in comparison to normals. (b). the cell population dynamics in draining lymph node using cell surface markers as indicated above in Specific aim #1. (c). the evolution of cytokine secretion by draining lymph node cells, both by direct ELISA assays, as well as semi-quantitative RT-PCR. 3. The role of nitric oxide in host defense against filarial infection. We will examine (a) the growth of B. malayi in mice in which the inducible nitric oxide synthase (iNOS) gene has been inactivated by homologous recombination. (b) the influence of alleles at the Nramp and iNOS loci on murine susceptibility to B. malayi. (c). the role of cytokines controlling the induction of nitric oxide in the resistance of NOD/LtSz-scid mice to B. malayi.