Chapoval Svetlana P, Iijima Koji, Marietta Eric V, Smart Michele K, Chapoval Andrei I, Andrews Amy G, David Chella S
Department of Immunology, Allergic Diseases Research Laboratory, and Section of Veterinary Medicine, Mayo Clinic, Rochester, MN 55905, USA.
J Immunol. 2002 Jan 15;168(2):890-9. doi: 10.4049/jimmunol.168.2.890.
To investigate the role of HLA-DQ molecules and/or CD4(+) T cells in the pathogenesis of allergic asthma, we generated HLA-DQ6 and HLA-DQ8 transgenic mice lacking endogenous class II (Abeta(null)) and CD4 genes and challenged them intranasally with short ragweed allergenic extract (SRW). We found that DQ6/CD4(null) mice developed a strong eosinophilic infiltration into the bronchoalveolar lavage and lung tissue, while DQ8/CD4(null) mice were normal. However, neither cytokines nor eosinophil peroxidase in the bronchoalveolar lavage of DQ6/CD4(null) mice was found. In addition, the airway reactivity to methacholine was elevated moderately in DQ6/CD4(null) mice compared with the high response in DQ/CD4(+) counterparts and was only partially augmented by CD4(+) T cell transfer. The DQ6/CD4(null) mice showed Th1/Th2-type cytokines and SRW-specific Abs in the immune sera in contrast to a direct Th2 response observed in DQ6/CD4(+) mice. The proliferative response of spleen mononuclear cells and peribronchial lymph node cells demonstrated that the response to SRW in DQ6/CD4(null) mice was mediated by HLA-DQ-restricted CD4(-)CD8(-)NK1.1(-) T cells. FACS analysis of PBMC and spleen mononuclear cells demonstrated an expansion of double-negative (DN) CD4(-)CD8(-)TCRalphabeta(+) T cells in SRW-treated DQ6/CD4(null) mice. These cells produced IL-4, IL-5, IL-13, and IFN-gamma when stimulated with immobilized anti-CD3. IL-5 ELISPOT assay revealed that DN T cells were the cellular origin of IL-5 in allergen-challenged DQ6/CD4(null) mice. Our study shows a role for HLA-DQ-restricted CD4(+) and DN T cells in the allergic response.
为了研究HLA - DQ分子和/或CD4(+) T细胞在过敏性哮喘发病机制中的作用，我们培育了缺乏内源性II类分子（Abeta(null)）和CD4基因的HLA - DQ6和HLA - DQ8转基因小鼠，并经鼻用短豚草变应原提取物（SRW）对它们进行攻击。我们发现，DQ6/CD4(null)小鼠支气管肺泡灌洗和肺组织出现强烈的嗜酸性粒细胞浸润，而DQ8/CD4(null)小鼠则正常。然而，在DQ6/CD4(null)小鼠的支气管肺泡灌洗中未发现细胞因子和嗜酸性粒细胞过氧化物酶。此外，与DQ/CD4(+)小鼠的高反应性相比，DQ6/CD4(null)小鼠对乙酰甲胆碱的气道反应性适度升高，并且仅部分地通过CD4(+) T细胞转移而增强。与DQ6/CD4(+)小鼠中观察到的直接Th2反应相反，DQ6/CD4(null)小鼠免疫血清中显示出Th1/Th2型细胞因子和SRW特异性抗体。脾单核细胞和支气管周围淋巴结细胞的增殖反应表明，DQ6/CD4(null)小鼠对SRW的反应由HLA - DQ限制的CD4(-)CD8(-)NK1.1(-) T细胞介导。对外周血单核细胞（PBMC）和脾单核细胞的流式细胞术分析表明，在经SRW处理的DQ6/CD4(null)小鼠中双阴性（DN）CD4(-)CD8(-)TCRalphabeta(+) T细胞扩增。当用固定化抗CD3刺激时，这些细胞产生IL - 4、IL - 5、IL - 13和IFN - γ。IL - 5酶联免疫斑点分析显示，在变应原攻击的DQ6/CD4(null)小鼠中，DN T细胞是IL - 5的细胞来源。我们的研究表明HLA - DQ限制的CD4(+)和DN T细胞在过敏反应中起作用。
