CHARACTERIZATION OF MOUSE NASAL LYMPHOCYTES ISOLATED BY ENZYMATIC EXTRACTION WITH COLLAGENASE
CHARACTERIZATION OF MOUSE NASAL LYMPHOCYTES ISOLATED BY ENZYMATIC EXTRACTION WITH COLLAGENASE
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DOI：
10.1016/0022-1759(95)00165-7
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发表时间：
1995-11-16
影响因子：
2.2
通讯作者：
TAMURA, SI
中科院分区：
文献类型：
--
作者：
ASANUMA, H;INABA, Y;TAMURA, SI
建立了一种分离小鼠鼻淋巴细胞的方法。通过胶原酶酶提取从解剖的小鼠鼻组织碎片中释放细胞，并通过逐步Percoll梯度离心分离。当用FACScan荧光分析仪分析时，来自正常BALB/c小鼠的部分纯化的鼻淋巴细胞组分含有CD 4（+）T细胞（18-23%）、CD 8（+）T细胞（7-10%）和B细胞（20-38%）。鼻细胞组分中T/B细胞和CD 4（+）/CD 8（+）T细胞的比例约为派尔集合淋巴结淋巴细胞的两倍。然后表征来自感染流感病毒的小鼠的鼻淋巴细胞部分。感染后7天，鼻淋巴细胞中CD 4（+）和CD 8（+）T细胞的数量分别是正常小鼠的2倍和3倍。它们产生IFN-γ，在感染后增加。它们含有在感染后4周分泌流感病毒特异性伊加和IgG抗体的细胞。此外，来自感染的C3 H小鼠的鼻淋巴细胞裂解病毒感染的靶细胞。这些结果表明，该方法可以成功地用于研究实验动物上呼吸道粘膜免疫的细胞动力学。
A reproducible method for isolation of mouse nasal lymphocytes was developed. The cells were released from tissue fragments of dissected mouse nose by enzyme extraction with collagenase and separated by a stepwise Percoll gradient centrifugation. The partially purified nasal lymphocyte fraction from normal BALB/c mice contained CD4(+) T cells (18-23%), CD8(+) T cells (7-10%) and B cells (20-38%), when analysed with a FACScan fluorescence analyser. The ratio of T to B cells and that of CD4(+) to CD8(+) T cells in the nasal cell fraction were about twice as high as those in Peyer's patch lymphocytes. The nasal lymphocyte fraction from the mice infected with influenza virus was then characterized. The nasal lymphocytes contained a twice larger number of CD4(+) and a three times larger number of CD8(+) T cells than those of normal mice 7 days after infection. They produced IFN-gamma which increased after infection. They contained the cells secreting influenza virus-specific IgA and IgG antibodies 4 weeks after infection. Moreover, the nasal lymphocytes from infected C3H mice lysed the virus infected-target cells. These results suggest that this method can successfully be used for investigating cellular dynamics of mucosal immunity in the upper respiratory tract of experimental animals.