新甲型H1N1流感重症死亡病例呼吸道及外周免疫器官病理形态学及免疫细胞的变化

目的 探索新型甲型H1N1流感病毒感染重症死亡病例呼吸道及外周免疫器官病理形态学及免疫细胞的变化。方法收集2009年北京甲型H1N1流感重症死亡病例8例,其中2例为系统尸体解剖标本,6例为床旁穿刺标本,以手足口病1例作为对照。HE染色观察病理形态学变化;免疫组织化学技术检测肺、脾和淋巴结免疫细胞变化。结果 实验组表现为坏死性支气管炎和周围炎,弥漫性肺损伤、肺出血、纤维化,巨噬细胞明显增生,淋巴细胞浸润不明显。甲型H1N1流感病毒血凝素和核蛋白抗原主要表达于呼吸道上皮及巨噬细胞。免疫细胞计数:CD68⁺巨噬细胞显著增多,CD20⁺B淋巴细胞、CD3⁺、CD4⁺、CD8⁺T细胞减少,CD56⁺细胞偶见,各类细胞与对照组比较差异无统计学意义。脾和淋巴结病变基本一致,灶状组织细胞增生,"噬红现象"明显,淋巴组织萎缩,残留滤泡内以滤泡树突状细胞(follicular dendritic cell, FDC)细胞为主。免疫细胞计数:CD68⁺巨噬细胞显著增多,但与对照组比较差异无统计学意义(P>0.05);CD20⁺B淋巴细胞、CD3⁺、CD4⁺、CD8⁺T免疫细胞数明显低于对照组(P<0.05);CD4⁺和CD8⁺T淋巴细胞的比值与对照组相比,差异无统计学意义;CD56⁺细胞在脾脏明显低于对照组,淋巴结内两组均偶见。结论 新型甲型H1N1流感重症患者外周免疫器官明显萎缩,特异性免疫功能减弱,其中细胞免疫反应下降更明显。

Pathological morphology change of respiratory and peripheral immune organ and the change of immune cells

Objective To explore the pathology morphological changes of trachea and peripheral immune organ and immune cells of the new influenza A (H1N1) virus severe infection. Methods To collect Collection of 2009 Beijing influenza A H1N1 intensive deaths 8 cases; Pathological changes of these tissue were observed by HE staining; The immune cell types were tested, quantity and distribution characteristics were made by immunohistochemistry. Results The principal pathologic changes of trachea were necrotizing bronchiolitis with peripheral inflammation, diffuse alveolar damage, pulmonary hemorrhage and fibrosis. Macrophages hyperplasia and little lymphocyte infiltration; Viral antigen was found in the epithelium of the epithelium and macrophage; CD68⁺ macrophage count significantly increased in lung, CD20⁺ B lymphocytes, CD3⁺, CD4⁺, CD8⁺ T count, CD56⁺ NK cells occasional,no obvious differences were found compared with the control group (P>0.05). Spleen and lymph nodes were same,focal histocyte hyperplasia, erythrocytophagy were observed, lymphoid tissue atrophy.IHC cell count showed CD68⁺ macrophage significantly increased, CD20⁺ B, CD3⁺, CD4⁺, CD8⁺ T lymphocytes were significantly lower than the control group (P<0.05). NK cells were rarely seen in the spleen and lymph nodes. The ratio of CD4⁺ and CD8⁺ T lymphocytes between experimental group and control group is not significant different. Conclusion The severe influenza A (H1N1) patients, peripheral immune organs showed significant atrophy, specific immune function was abated and the cellular immune response decreased more significantly.