An Investigation of Mucosal Immunity in Nasopharynx

为了更好地了解鼻腔免疫应答的机制，本研究以C57BL/6和ova -转基因（OVA23-3）小鼠为实验对象，用霍乱毒素（CT）对其进行反复鼻腔抗原刺激，研究其鼻腔黏膜免疫的变化。针对相应HRP或OVA抗原的ag特异性IgA抗体滴度在ct处理小鼠的鼻洗液中增加。在我们的免疫化学实验中，在洗鼻液中具有IgA Ab活性的小鼠鼻黏膜中发现了携带IgA的B细胞。在FACS分析中，我们收集了CD3+T细胞和B细胞，在鼻黏膜中发现了比盐水挑战小鼠更多的α - TCR+ T细胞。γ δ TCR+ T细胞也被发现。用密度仪对细胞因子基因表达进行半定量分析，发现ct处理小鼠鼻淋巴细胞中ifn - γ、IL-2和IL-6的表达明显强于盐水刺激小鼠。这些结果可能表明α - TCR+和γ - TCR+ T细胞在鼻咽部B细胞的组装和调节中发挥重要作用。我们成功地将人扁桃体淋巴细胞转移到NOD-scid小鼠体内，以检验扁桃体淋巴细胞在鼻咽和管腔等外周粘膜部位作为免疫反应诱导部位的确切作用。人白细胞（CD45+）、T细胞（CD3+）和B细胞（CD20+）被移植到脾脏和其他非淋巴器官。流式细胞术分析显示，在腹腔内转移4周后，NOD-scid小鼠脾脏中检测到不同程度的人CD45+、CD3+、CD4+（辅助/诱导剂）、CD8+（抑制/细胞毒性）细胞。然而，在CB-17 scid小鼠中未见移植。免疫染色显示，人白细胞存在于脾脏、肺和肝脏，但不存在于正常的鼻咽或管鼻粘膜。将α链球菌携带者的人扁桃体淋巴细胞植入NOD-scid小鼠，检测小鼠血清中m蛋白特异性人IgG和IgA Ab的活性。用CT对这些小鼠进行m蛋白鼻内免疫后，鼻洗液中m蛋白特异性人IgA抗体滴度高于未经m蛋白鼻内攻击的小鼠。在免疫染色中，人T细胞和B细胞被募集到鼻咽淋巴网状组织（NALT）周围的鼻黏膜，这些小鼠具有较高的人IgA Ab滴度。这些结果可能使我们将该模型作为一种有希望的工具，用于评估淋巴细胞归巢和募集前体细胞淋巴细胞最终分化的机制，以建立上呼吸道外周粘膜部位的抗原特异性免疫反应。少

For better understanding mechanism of nasal immune response, mucosal immunity of nasal mucosa was investigated in C57BL/6 and OVA-transgenic (OVA23-3) mice stimulated with repeated nasal challenge of antigen with cholera toxin (CT). Ag-specific IgA Ab titers against corresponding HRP or OVA antigens increased in nasal washings of CT-treated mice. In our immunochemistry, IgA-bearing B cells were found in nasal mucosa of mice with IgA Ab activity in nasal washings. In FACS analysis, CD3+T cells as well as B cells were harvested and alphabeta TCR+ T cells are found in a higher number in nasal mucosa, than saline-challenged control mice. gammadelta TCR+ T cells were also found. Semiquantitative analysis of cytokine gene expression by densitometer demonstrates that the expression of IFN-gamma, IL-2, and IL-6 was significaytly stronger in nasal lymphocytes of CT-treated mice, than in those of saline-challenged mice. These results might indicate that alphabeta TCR+ and gammadeltaTCR+ T cells … More play an important role for mounting and regulating IgA production of B cells in nasopharynx. A transfer study of human tonsillar lymphocytes into NOD-scid mice were successfully conducted in roder to examine the exact role of tonsilar lymphocytes as an inductive site of the immune response in peripheral mucosal sites such as nasopharynx and tubotympanum. Human leukocytes (CD45+), T cells (CD3+), and B cells (CD20+) are engrafted in the spleens and otehr non-lymphoid organ. In a flow cytometric analysis, human CD45+, CD3+, CD4+ (helper/inducer), CD8+ (suppressor/cytotoxic) cells were detected in various degrees in the spleens of NOD-scid mice at a 4-week interval after the intraperitoneal transfer. However, no engrAftment was seen in CB-17 scid mice. An immunostaining revealed that human leukocytes were present in the spleen, lung, and liver, but not in normal nasopharyngeal or tubotympanal mucosae. In NOD-scid mice engrafted with human tonsillar lymphocytes obtained from patients carrying alpha Streptococci, M-protein specific human IgG and IgA Ab activities were detected in the sera of these mice. Upon an intranasal immunization of M-protein with CT to these mice, M-protein specific human IgA antibody titers in nasal washings were higher than those of mice without the intransal challenge of M-protein. In an immunostaining, human T and B cells were recruited to the nasal mucosae around the nasopharyngeal lymphoreticular tissue (NALT) of those mice having higher human IgA Ab titers. These results may allow us to use this model as a promising tool for evaluating the lymphocyte homing and mechanism of the final differentiation of recruiting precurosr lymphocytes for mounting antien-specific immune response in peripheral mucosal sites of upper respiratory tract. Less