Laser-capture microdissection of plasma cells from subacute sclerosing panencephalitis brain reveals intrathecal disease-relevant antibodies
Laser-capture microdissection of plasma cells from subacute sclerosing panencephalitis brain reveals intrathecal disease-relevant antibodies
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DOI：
10.1073/pnas.0502323102
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发表时间：
2005-05-17
影响因子：
11.1
通讯作者：
Gilden, DH
中科院分区：
文献类型：
--
作者：
Burgoon, MP;Keays, KM;Gilden, DH
在患有慢性传染性CNS疾病的人的脑脊液中发现IgG和寡克隆带增加。研究表明，这些寡克隆带是针对致病因子的抗体。采用激光捕获显微切割技术从亚急性硬化性全脑炎患者的脑中分离单个CD38+浆细胞，并采用单细胞RT-PCR分析每个细胞表达的单个IgG重链和轻链。基于过度代表的IgG序列，我们构建了功能性重组抗体（重组IgG），并确定其特异性。八分之五的重组IgG识别麻疹病毒，这是亚急性硬化性全脑炎的原因。这些结果表明，在死后的大脑中，过表达的IgG序列可用于产生识别其靶抗原的功能性重组抗体。该策略可用于鉴定病因不明的CNS炎性疾病中的疾病相关抗原。
Increased IgG and oligoclonal bands are found in cerebrospinal fluid of humans with chronic infectious CNS disease. Studies have shown that these oligoclonal bands are antibodies directed against the agent that causes disease. Laser-capture microdissection was used to isolate individual CD38+ plasma cells from the brain of a patient with subacute sclerosing panencephalitis, and single-cell RT-PCR was used to analyze individual IgG heavy and light chains expressed by each cell. Based on overrepresented IgG sequences, we constructed functional recombinant antibodies (recombinant IgGs) and determined their specificities. Five of eight recombinant IgGs recognized measles virus, the cause of subacute sclerosing panencephalitis. These results demonstrate that overrepresented IgG sequences in postmortem brains can be used to produce functional recombinant antibodies that recognize their target antigens. This strategy can be used to identify disease-relevant antigens in CNS inflammatory diseases of unknown etiology.