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- 2018
TIA重组cDNA表达克隆血清抗原基因的识别
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Abstract:
摘要 目的:应用重组cDNA表达克隆血清学鉴定技术(SEREX)筛选与识别短暂性脑缺血发作(TIA)血清相关性抗原基因.方法:采集19例TIA患者血清,免疫筛选人主动脉内皮细胞cDNA表达文库;对阳性靶抗原行单克隆扩增,经单克隆内切除后转换载体pBluescript SK(+)提取质粒;对包含cDNA插入目的基因片段的质粒进行DNA序列检测,通过NCBI-BLAST同源性检索,鉴定具有免疫原性的独立的TIA重组cDNA表达克隆血清抗原基因.结果:(1) SEREX筛选出172个重组cDNA阳性单克隆;(2)单克隆内切除后获得167个pBluescript质粒,酶切鉴定质粒中均包含有目的基因插入片段;(3)生物信息学分析:识别11个独立的重组cDNA表达克隆抗原基因.(4)基因功能解析:发现matrix metalloproteinase 1 (MMP1)、chromobox homolog 1 (CBX1)和chromobox homolog 5 (CBX5) 与TIA和脑梗死(CI)的发生、发展关系密切.结论:应用SEREX技术成功筛选及鉴定出TIA相关靶抗原基因,发现MMP1、CBX1和CBX5可能与短暂性脑缺血发作(TIA)和脑梗死(CI)发生发展有关.
| [1] | WANG W,JIANG B,SUN H, et al.Prevalence, incidence,and mortality of stroke in China clinical perspective: results from a nationwide population-based survey of 480,687 adults[J].Circulation,2017,135(8):759. |
| [2] | GILES M F,ROTHWELL P M.Risk of stroke early after transient ischaemic attack:a systematic review and meta-analysis[J].Lancet Neurol,2007,6(12):1063-1072. |
| [3] | GOMEZ D,SWIATLOWSKA P,OWENS G K.Epigenetic control of smooth muscle cell identity and lineage memory[J].Arterioscler Thromb Vasc Biol,2015,35(12):2508-2516. |
| [4] | JOHNSTON S C.Ischemic preconditioning from transient ischemic attacks? Data from the Northern California TIA Study[J].Stroke,2004,35(11 Suppl 1):2680-2682. |
| [5] | SAHIN U,TURECI O,SCHMITT H, et al.Human neoplasms elicitmultiple specific immune responses in the autologous host[J].Proc Natl Acad Sci USA,1995, 92(25):11810-11813. |
| [6] | LEE S Y,JEOUNG D.The reverse proteomics for identification of tumor antigens[J].J Microbiol Biotechnol,2007,17(6):879-890. |
| [7] | HE R,YANG N,ZHANG P, et al.Identification and expression of MMSA-8, and its clinical significance in multiple myeloma[J].Oncology Reports,2017,37(6):3235-3243. |
| [8] | MUTO M,MORI M,LIU J,et al.Serum soluble Talin-1 levels are elevated in patients with multiple sclerosis,reflecting its disease activity[J].Journal of Neuroimmunology,2017,305:131-134. |
| [9] | MATSUURA E,ATZENI F,SARZI-PUTTINI P,et al.Is atherosclerosis an autoimmune disease[J].BMC Med.2014,12:47. |
| [10] | NAGASE H,BARRETT A J,JR W J.Nomenclature and glossary of the matrix metalloproteinases[J].Matrix Supplement,1992,1:421. |
| [11] | MAYMON E,ROMERO R,PACORA P, et al.Evidence for the participation of interstitial collagenase (matrix metalloproteinase 1) in preterm premature rupture of membranes[J].Am J Obstet Gynecol,2000,183:914-920. |
| [12] | UCHIDA K,AKITA Y,MATSUO K,et al.Identification of specific auto-antigens in Sjogren's syndrome by SEREX[J].Immunology,2005,116(1):53-63. |
| [13] | QIN W,FENG S G,DENG H X,et al.Serological screening of autoantigens associated with vasculitis in systemic lupus erythematosus[J].Journal of Sichuan University,2007,38(1):132-134. |
| [14] | GOTO M,KURIBAYASHI K,TAKAHASHI Y, et al.Identification of autoantibodies expressed in acquired aplastic anaemia[J].Br J Haematol,2013,160(3):359-362. |
| [15] | MUTO M,MORI M,HIWASA T, et al.Novel serum autoantibodies against talin1 in multiple sclerosis:Possible pathogen tic roles of the antibodies[J].J Neuroimmunol,2015,284:30-36. |
| [16] | HAO S,FU R,WANG H, et al.Screening novel autoantigens targeted by serum IgG autoantibodies in immunorelated pancytopenia by SEREX[J].International Journal of Hematology,2017. |
| [17] | OSHIMA J,HISAMA F M.Search and insights into novel genetic alterations leading to classical and atypical Werner syndrome[J].Gerontology,2014,60(3):239-246. |
| [18] | WILLIAMS P A,SILVA E A.The role of synthetic extracellular matrices in endothelial progenitor cell homing for treatment of vascular disease[J].Ann Biomed Eng,2015,43:2301-2313. |
| [19] | AMIN M,PUSHPAKUMAR S,MURADASHVILI N,et al.Regulation and involvement of matrix metalloproteinases in vascular diseases[J].Front Biosci(Landmark Ed),2016,21:89-118. |
| [20] | HAO Y,TIAN S,SUN M, et al.Association between matrix metalloproteinase gene polymorphisms and development of ischemic stroke[J].Int J Clin Exp Pathol, 2015,8(9):11647-11652. |
| [21] | LOMBERK G,WALLRATH LL,URRUTIA R.The heterochromatin protein 1 family[J]. Genome Biol,2006,7:228. |
| [22] | EISSENBERG J C,ELGIN S C.HP1a:A structural chromosomal protein regulating transcription[J].Trends Genet,2014,30(3):103-110. |
| [23] | MOTEGI S,YOKOYAMA Y,UCHIYAMA A, et al.First Japanese case of atypical progeroid syndrome/atypical Werner syndrome with heterozygous LMNA mutation[J].J Dermatol,2014,41(12):1047-1052. |
