Detection of autoantibodies to ss‐a/ro by indirect immunofluorescence using a transfected and overexpressed human 60 kd ro autoantigen in hep‐2 cells

Journal of Clinical Laboratory Analysis - Tập 9 Số 3 - Trang 218-224 - 1995
Marvin J. Fritzler1, B. Joan Miller1
1McCaig Center for Joint Injury and Arthritis Research, Faculty of Medicine, University of Calgary, Calgary, Canada

Tóm tắt

AbstractThe objective of the study was to determine the sensitivity and specificity of an indirect immunofluorescence (IIF) assay using transfected HEp‐2 cells to detect anti‐SS‐A/Ro autoantibodies in human sera. Seventy‐three sera having SS‐A/Ro autoantibodies as determined by double immunodiffusion (ID) and immunoblotting (IB) were tested by IIF on a HEp‐2 cell substrate that had been transfected with a full‐length cDNA encoding a human 60 kD SS‐A/Ro autoantigen. Controls included 30 normal human sera and 50 sera with a variety of other antinuclear antibodies. Prototype human and rabbit sera directed against the 60 kD SS‐A/Ro antigen produced intense speckled nuclear and nucleolar staining of transfected cells. Sixty‐nine of 73 (95%) SS‐A/Ro positive sera also produced this characteristic staining pattern. The endpoint autoantibody titers on transfected cells was fivefold greater than on untransfected cells. The 30 normal human sera and the 50 sera with other antinuclear antibodies did not produce this characteristic staining. Six of 32 (19%) unselected sera that were sent for autoantibody testing had reactivity with transfectants by IIF. Four of the six sera were confirmed to have anti‐SS‐A/Ro antibodies by ID and 5/6 by IB. By contrast, only three of these sera were scored as having a staining pattern compatible with SS‐A/Ro antibodies by IIF on standard HEp‐2 substrates. We conclude that SS‐A/Ro autoantibodies can be detected by an IIF assay using a HEp‐2 cell substrate transfected with a SS‐A/Ro cDNA. This new substrate detects SS‐A/Ro antibodies that were not identified on standard HEp‐2 substrates and by other immunoassays.©1995 wiley‐Liss, inc.

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Tài liệu tham khảo

10.1016/S0140-6736(61)92430-8

10.4049/jimmunol.102.1.117

10.1172/JCI108007

10.1002/art.1780220719

10.1126/science.6164096

Slobbe RL, 1991, Ro (SS‐A) and La (SS‐B) ribonucleoprotein complexes: Structure, function and antigenicity, Ann Med Interne (Paris), 142, 592

Chan EKL, 1992, Antinuclear antibodies in Sjögren's syndrome, Rheum Dis Clin North Am, 18, 551, 10.1016/S0889-857X(21)00316-1

10.1093/rheumatology/32.5.396

10.1007/BF00986765

10.1172/JCI114704

Rokeach LA, 1991, Characterization of the autoantigen calreticulin, J Immunol, 147, 3031, 10.4049/jimmunol.147.9.3031

10.1002/art.1780270207

10.1172/JCI112892

Xia P, 1987, The particulate (speckled‐like thread) nuclear staining pattern: Species and cellular distribution of the Ro/SS‐A antigen, J Clin Lab Immunol, 22, 101

Takano S, 1989, Detection of anti‐SS‐A/Ro antibody by using pig spleen supernatant and characterization of corresponding antigen, Jap J Rheumatol, 2, 67

10.1073/pnas.81.7.1996

10.1002/art.1780281207

10.1084/jem.167.5.1560

10.1172/JCI114014

10.1016/0022-1759(90)90440-7

10.1111/j.1365-2249.1991.tb05780.x

10.1093/rheumatology/32.6.449

10.1016/0022-1759(90)90362-Y

10.1172/JCI114551

10.1002/art.1780340613

ChanEKL BuyonJP:The SS‐A/Ro antigen. In Manual of Biological Markers of Disease.Wvan ven Roij TMaini eds B4:1994.

10.1128/MCB.1.12.1138

10.1073/pnas.81.3.659

10.1006/clin.1994.1181

Chan EKL, 1992, Manual Clin Lab Immunol, 755

10.1016/0090-1229(85)90045-5

10.1016/0161-5890(90)90079-F

10.1172/JCI115003

10.1111/j.1462-5822.2007.00901.x

10.1073/pnas.76.9.4350

Fritzler MJ, 1986, Methods and Achievements in Experimental Pathology, 225

Fritzler MJ, 1985, Laboratory Diagnostic Procedures in the Rheumatic Diseases, 207

10.1002/art.1780190214

10.1016/0090-1229(89)90158-X

10.1172/JCI117145

10.1097/00005792-196505000-00001

10.1016/0896-8411(89)90167-4

Fritzler MJ, 1986, The detection of autoantibodies on HEp‐2 cells using an indirect immunoperoxidase kit (Colorzyme), Diagnostic Immunology, 4, 217

10.1016/S0065-2776(08)60641-0

Fritzler MJ, 1992, Systemic Lupus Erythematosus Systemic Lupus Erythematosus, 273

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Thông tin xuất bản

Journal of Clinical Laboratory Analysis

Tập 9 Số 3

218-224

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