Owens Gregory P, Ritchie Alanna M, Burgoon Mark P, Williamson R Anthony, Corboy John R, Gilden Donald H
Department of Neurology, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Mail Stop Box B182, Denver, CO 80262, USA.
J Immunol. 2003 Sep 1;171(5):2725-33. doi: 10.4049/jimmunol.171.5.2725.
Single-cell RT-PCR was used to sample CD19(+) B cell repertoires in cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) or viral meningitis. Analysis of amplified Ab H and L chain products served to identify the rearranged germline segment and J segment, and to determine the degree of homology for the H and L chain sequence of individual B cells. The B cell repertoire of viral meningitis CSF was predominantly polyclonal, whereas B cell clonal expansion was a prominent feature of the IgG repertoire in three of four MS patients. Two dominant clonal populations in one MS CSF accounted for approximately 70% of the IgG H chain V regions sequenced, while the corresponding IgM repertoires were more heterogeneous. One clonal B cell population revealed multiple L chain rearrangements, raising the possibility of a role for receptor editing in shaping the B cell response in some MS patients. The most immediate implications of identifying rearranged Ig sequences in MS B cells is the potential to accurately recreate recombinant Abs from these overrepresented H and L chains that can be used to discover the relevant Ag(s) in MS.
采用单细胞逆转录聚合酶链反应（RT-PCR）对多发性硬化症（MS）或病毒性脑膜炎患者脑脊液（CSF）中的CD19（+）B细胞库进行采样。对扩增的抗体重链（H）和轻链（L）产物进行分析，以鉴定重排的胚系区段和连接区段（J区段），并确定单个B细胞H链和L链序列的同源程度。病毒性脑膜炎脑脊液的B细胞库主要为多克隆性，而在4例MS患者中的3例，B细胞克隆性扩增是IgG库的一个突出特征。1例MS患者脑脊液中的两个优势克隆群体约占所测序IgG H链V区的70%，而相应的IgM库则更为异质性。一个克隆性B细胞群体显示出多个L链重排，这增加了受体编辑在某些MS患者B细胞反应形成中发挥作用的可能性。在MS患者B细胞中鉴定重排的Ig序列最直接的意义在于，有可能从这些过度表达的H链和L链中准确重建重组抗体，从而用于发现MS中的相关抗原。
