IDENTIFICATION OF NEW B-CELL EPITOPES IN THE SERA OF RHEUMATOID-ARTHRITIS PATIENTS USING A RANDOM NANOPEPTIDE PHAGE LIBRARY

被引:52
作者
DYBWAD, A
FORRE, O
KJELDSENKRAGH, J
NATVIG, JB
SIOUD, M
机构
[1] INST IMMUNOL & RHEUMATOL,OSLO,NORWAY
[2] OSLO SANITETSFORENING REUMATISM HOSP,OSLO,NORWAY
关键词
RHEUMATOID ARTHRITIS; EPITOPE; LIBRARY;
D O I
10.1002/eji.1830231222
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
A random nanopeptide phage library was used to screen a pool of immunoglobulin fractions obtained from rheumatoid arthritis (RA) patients. After three rounds of panning, random individual phages were selected by their capacity to react with individual sera from RA patients. By sequencing the inserts corresponding to the peptides displayed on the surface of the phages, we found that phages displaying particular peptides were overrepresented in the selected libraries. The peptides displayed by these phages were: pep1 = Ala-Asp-Gly-Gly-Ala-Gln-Gly-Thr-Ala; pep2 = Pro-Gly-Pro-Ser-Arg-Ala-His-Phe-Leu; pep3 = Leu-Ser-Ser-Arg-Glu-Pro-Gln-Ala-Arg; pep4 = Arg-Leu-Thr-Arg-Glu-Leu-Tyr-Ala-Gln and pep5 = Tyr-Thr-Gln-Lys-His-Gln-Ala. The percentage of sera positive for pep1 was higher in RA patients as compared to the normal adults (p < 0.0004) and the reacting antibody was mainly of IgG isotype. The specificity of binding to the phage displaying pep1 was confirmed by competition experiments using both isolated phages and a synthetic peptide. Interestingly, a mutated phage displaying only Ala-Asp-Gln-Gly-Thr-Ala had no significant reactivity with the sera, indicating that the amino acids (Gly-Gly-Ala) of pep1 are the vital for the binding.Taken together this study demonstrates that it is possible to select specific ligands from a random phage library using sera from RA patients. In addition, this approach could be useful for identifying peptide antigens that might be part of causitive agents in autoimmune diseases.
引用
收藏
页码:3189 / 3193
页数:5
相关论文
共 23 条
[1]   ELEVATED LEVELS OF ANTIBODIES TO EPSTEIN-BARR VIRUS-ANTIGENS IN SERA AND SYNOVIAL-FLUIDS OF PATIENTS WITH RHEUMATOID-ARTHRITIS [J].
ALSPAUGH, MA ;
HENLE, G ;
LENNETTE, ET ;
HENLE, W .
JOURNAL OF CLINICAL INVESTIGATION, 1981, 67 (04) :1134-1140
[2]  
CASSIDY JT, 1989, B RHEUM DIS, V38, P1
[3]   PEPTIDES ON PHAGE - A VAST LIBRARY OF PEPTIDES FOR IDENTIFYING LIGANDS [J].
CWIRLA, SE ;
PETERS, EA ;
BARRETT, RW ;
DOWER, WJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (16) :6378-6382
[4]  
DELVIN JJ, 1990, SCIENCE, V249, P404
[5]   SELECTION OF ANTIBODY LIGANDS FROM A LARGE LIBRARY OF OLIGOPEPTIDES EXPRESSED ON A MULTIVALENT EXPOSITION VECTOR [J].
FELICI, F ;
CASTAGNOLI, L ;
MUSACCHIO, A ;
JAPPELLI, R ;
CESARENI, G .
JOURNAL OF MOLECULAR BIOLOGY, 1991, 222 (02) :301-310
[6]   RECOGNITION OF A MYCOBACTERIA-SPECIFIC EPITOPE IN THE 65-KD HEAT-SHOCK PROTEIN BY SYNOVIAL FLUID-DERIVED T-CELL CLONES [J].
GASTON, JSH ;
LIFE, PF ;
JENNER, PJ ;
COLSTON, MJ ;
BACON, PA .
JOURNAL OF EXPERIMENTAL MEDICINE, 1990, 171 (03) :831-841
[7]   MECHANISMS OF ANTIBODY-BINDING TO A PROTEIN [J].
GETZOFF, ED ;
GEYSEN, HM ;
RODDA, SJ ;
ALEXANDER, H ;
TAINER, JA ;
LERNER, RA .
SCIENCE, 1987, 235 (4793) :1191-1196
[8]   SEQUENCE HOMOLOGIES BETWEEN HSP60 AND AUTOANTIGENS [J].
JONES, DB ;
COULSON, AFW ;
DUFF, GW .
IMMUNOLOGY TODAY, 1993, 14 (03) :115-118
[9]   THE IMPORTANCE OF THE T-CELL IN INITIATING AND MAINTAINING THE CHRONIC SYNOVITIS OF RHEUMATOID-ARTHRITIS [J].
PANAYI, GS ;
LANCHBURY, JS ;
KINGSLEY, GH .
ARTHRITIS AND RHEUMATISM, 1992, 35 (07) :729-735
[10]   ANTIBODY-SELECTABLE FILAMENTOUS FD PHAGE VECTORS - AFFINITY PURIFICATION OF TARGET GENES [J].
PARMLEY, SF ;
SMITH, GP .
GENE, 1988, 73 (02) :305-318