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目的探讨变应性鼻炎免疫治疗的新方法。方法构建细胞毒性T淋巴细胞相关抗原4(cytotoxic T lymphocyteassoc iated antigen4,CTLA4)胞外区蛋白的酵母表达系统,以获取具有生物学活性的CTLA4胞外区蛋白;采用卵清蛋白(ovalbumin,OVA)致敏和激发诱导小鼠变应性鼻炎。CTLA4胞外区蛋白组在每次激发前予以CTLA4胞外区蛋白200μg/只腹腔注射,观察小鼠变应性鼻炎相关症状和鼻黏膜形态学改变。结果通过CTLA4胞外区蛋白的酵母表达系统可获得相对分子质量为28000、经蛋白免疫印迹(Westernblot)鉴定的CTLA4胞外区蛋白。该纯化蛋白能显著抑制混合淋巴细胞反应中T细胞的增殖,抑制率为95.4%。变应性鼻炎组小鼠在OVA激发后出现流涕、喷嚏等症状;鼻黏膜出现组织水肿、充血、炎性细胞渗出等形态学改变。CTLA4胞外区蛋白组小鼠鼻部症状不明显,鼻黏膜无明显形态学改变。结论CTLA4胞外区蛋白可阻断小鼠变应性鼻炎的发生,其作用机理可能与其抑制T细胞活化有关。
Objective To explore a new method of immunotherapy for allergic rhinitis. Methods The yeast expression system of extracellular domain of cytotoxic T lymphocyteassociated antigen 4 (CTLA4) was constructed to obtain the biological activity of extracellular domain of CTLA4. Ovalbumin (OVA) Sensitization and challenge induced allergic rhinitis in mice. CTLA4 extracellular region of the protein group before each challenge CTLA4 extracellular protein 200μg / only intraperitoneal injection to observe the symptoms of allergic rhinitis and nasal mucosal morphological changes in mice. Results The CTLA4 extracellular region protein was identified by Western blot in a yeast expression system of CTLA4 extracellular domain. The purified protein could significantly inhibit the proliferation of T cells in mixed lymphocyte reaction with the inhibition rate of 95.4%. Allergic rhinitis group mice after OVA challenge runny nose, sneezing and other symptoms; nasal mucosa tissue edema, congestion, inflammatory cell exudation and other morphological changes. The nasal symptoms of CTLA4 extracellular protein group mice were not obvious, and there was no obvious morphological changes in nasal mucosa. Conclusion CTLA4 extracellular domain protein can block the occurrence of allergic rhinitis in mice and its mechanism may be related to the inhibition of T cell activation.