基于SEREX技术筛选的自身抗原FTL在免疫相关性全血细胞减少症中的作用研究

In our previous studies ,we had indicated that Immune-related pancytopenia (IRP) is a kind of bone marrow failure syndromes，which is caused by B cell hyperfunction and mediated by autoantibodies.And we had screened ferritin light chain(FTL) targeted by autoantibodies by SEREX. Thus, we hypothesize that FTL maybe the autoantigen of IRP , it is presented to Th2 cells by B lymphocytes ,then B lymphocytes are activated and produce autoantibodyies.Based on the hypothesis，we intend to find the epitope of FTL by constructing and screening FTL peptide library and identify the antigen peptide recognized by Th2 cells by ELIspot;then we detected FTL protein、FTL-mRNA、activating molecule of B cells、antigen auto－reactive T cells、anti-FTL antibodies by FCM、WB、ELISA、RT-PCR、MHCII－peptide tetramer－FCM and so on,meanwhile we compare their difference between case group and control group.At last,we construct IRP murine node linduced by antigen peptide.In conclusion,our group will expore the etiology of IRP to be the basis of further study,provideng a new diagnostic or therapeutic target for IRP.

本课题组前期研究证明免疫相关性全血细胞减少症（Immune-related pancytopenia, IRP）是一类B淋巴细胞功能亢进、分泌自身抗体介导骨髓衰竭所致的全血细胞减少症，且已用SEREX法筛选出能被自身抗体所识别的靶抗原铁蛋白轻链（Ferritin light chain,FTL ），因此提出假设：FTL可能为IRP的自身抗原，它被B淋巴细胞递呈给Th2细胞进而激活B淋巴细胞产生抗FTL自身抗体。本课题我们构建FTL肽库，寻找FTL的抗原表位，ELIspot筛选能被Th2细胞识别的FTL抗原肽；运用流式细胞术、WB、ELISA、RT-PCR、MHCII-肽四聚体流式细胞术等检测FTL水平、B淋巴细胞共刺激分子、抗原自身反应性T淋巴细胞、抗FTL抗体在病例组和对照组的差异；抗原肽诱导构建IRP小鼠模型。本课题为进一步揭示IRP病因奠定基础，提供IRP诊断及治疗的新靶点。

本课题组前期研究证明免疫相关性全血细胞减少症（Immune-related pancytopenia, IRP）是一类B淋巴细胞功能亢进、分泌自身抗体介导骨髓衰竭所致的全血细胞减少症，且已用SEREX法筛选出能被自身抗体所识别的靶抗原铁蛋白轻链（Ferritin light chain,FTL ），因此提出假设：FTL可能为IRP的自身抗原，它被B淋巴细胞递呈给Th2细胞进而激活B淋巴细胞产生抗FTL自身抗体。本课题我们通过噬菌体随机肽库寻找FTL的抗原表位，ELIspot筛选能被Th2细胞识别的FTL抗原肽；运用流式细胞术、WB、ELISA、RT-PCR、MHCII-肽四聚体流式细胞术等检测FTL水平、B淋巴细胞共刺激分子、抗原自身反应性T淋巴细胞、抗FTL抗体在病例组和对照组的差异。本课题为进一步揭示IRP病因奠定基础，提供IRP诊断及治疗的新靶点。