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目的:观察重组Hap蛋白免疫抗不可分型流感嗜血杆菌(NHTi)感染的免疫保护效果,初步探讨其保护机制。方法:用纯化的Hap重组蛋白与黏膜免疫佐剂霍乱肠毒素B亚单位(CT-B)联合鼻腔免疫C57BL/6小鼠。NHTi琼脂珠悬液对免疫小鼠进行气管内注射,建立小鼠NHTi急性肺部感染模型。NHTi攻击5d后,收集实验小鼠的支气管肺泡灌洗液(BALF)进行白细胞计数,并取其肺组织进行肺部荷菌数检测及肺组织的病理检测。结果:Hap重组蛋白与CT-B的联合免疫能显著降低小鼠BALF中的白细胞总数、中性粒细胞渗出(P<0.05)及感染小鼠肺组织内NTHi的荷菌数,并减轻感染小鼠支气管、细支气管周围及肺泡内的炎性浸润程度,明显改善其肺组织的病变。结论:鼻腔免疫的Hap重组蛋白在动物体内能发挥良好抗NTHi感染的免疫保护作用,为NTHi新型疫苗的研发提供了理论基础和实验依据。
Objective: To observe the immunoprotective effect of recombinant Hap protein against non-typeable Haemophilus influenzae (NHTi) infection and to explore its protective mechanism. Methods: C57BL / 6 mice were immunized with purified Hap recombinant protein and mucosal immune adjuvant Cholera toxin B subunit (CT-B) combined with nasal cavity. NHTi agar beads suspension of mice immunized intratracheal injection to establish a mouse model of acute lung infection in NHTi. After 5 days of NHTi challenge, BALB / c mice were collected for leukocyte counting and the lung tissues were taken for the detection of the number of lungs and the pathological examination of lung tissue. Results: Combined immunization with Hap recombinant protein and CT-B could significantly reduce the total number of leukocytes, neutrophil exudation (P <0.05) and the number of NTHi-infected mice in BALF and reduce the infection Mice bronchial, bronchial bronchial and alveolar inflammatory infiltration within the extent of significant improvement in the lung tissue lesions. Conclusion: Nasal immunization of Hap recombinant protein can play a good immune protection against NTHi infection in animals, which provides a theoretical basis and experimental basis for the development of a novel NTHi vaccine.