Abstract
A murine hybridoma cell line (aDNA35I9) secreting anti-DNA antibodies was used as a model for haemolytic plaque forming cells in an assay where DNA-conjugated sheep red blood cells (DNA-SRBC) were used as target cells. DEAE-dextran, employed to abolish the anti-complementary activity of agar gel, completely inhibited anti-DNA plaques. This problem was solved by using agarose instead of agar. Since the occurrence of small plaques may make reading of the test difficult, it was established that plaque size could be increased by decreasing the antigen density on the target cells. Free DNA in the gel inhibited plaque formation, indicating the specificity of the assay. Spleen cells from mice of the strains MRL/MP and NZB/W which are known to develop a stage of autoimmunity, produced plaques in numbers which were correlated to the age of the mice and to the anti-DNA antibody level in serum.
Full text
PDF




Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aarden L. A., Lakmaker F., Feltkamp T. E. Immunology of DNA. I. The influence of reaction conditions on the Farr assay as used for the detection of anti-ds DNA. J Immunol Methods. 1976;10(1):27–37. doi: 10.1016/0022-1759(76)90004-1. [DOI] [PubMed] [Google Scholar]
- Aarden L. A., de Groot E. R., Feltkamp T. E. Immunology of DNA. III. Crithidia luciliae, a simple substrate for the determination of anti-dsDNA with the immunofluorescence technique. Ann N Y Acad Sci. 1975 Jun 30;254:505–515. doi: 10.1111/j.1749-6632.1975.tb29197.x. [DOI] [PubMed] [Google Scholar]
- Agger R., Petersen J., Dinesen B., Wiik A., Andersen V. Production and secretion of immunoglobulins by in vitro-activated human B lymphocytes. Allergy. 1982 Apr;37(3):179–185. doi: 10.1111/j.1398-9995.1982.tb01894.x. [DOI] [PubMed] [Google Scholar]
- Alarcón-Segovia D. The role of the laboratory in rheumatology. Antibodies to nuclear and other intracellular antigens in the connective tissue diseases. Clin Rheum Dis. 1983 Apr;9(1):161–175. [PubMed] [Google Scholar]
- Aotsuka S., Okawa M., Ikebe K., Yokohari R. Measurement of anti-double-stranded DNA antibodies in major immunoglobulin classes. J Immunol Methods. 1979;28(1-2):149–162. doi: 10.1016/0022-1759(79)90337-5. [DOI] [PubMed] [Google Scholar]
- Bastian D., Borel H., Sasaki T., Steinberg A. D., Borel Y. Immune response to nucleic acid antigens and native DNA by human peripheral blood lymphocytes in vitro. J Immunol. 1985 Sep;135(3):1772–1777. [PubMed] [Google Scholar]
- Bell D. A., Clark C., Blomgren S. E., Vaughan J. H. Anti DNA antibody production by lymphoid cells of NZB-W mice and human systemic lupus erythematosus (SLE). Clin Immunol Immunopathol. 1973 Apr;1(3):293–303. doi: 10.1016/0090-1229(73)90046-9. [DOI] [PubMed] [Google Scholar]
- Clark C., Bell D. A., Vaughan J. H. Anti-DNA plaque-forming cells (PFC) and rosette-forming cells (RFC) in spleens of NZB-WF female (B-W) mice. J Immunol. 1972 Nov;109(5):1143–1145. [PubMed] [Google Scholar]
- Granholm N. A., Graves K., Cavallo T. Influence of avidity of circulating anti-DNA antibodies on murine lupus nephritis. J Clin Lab Immunol. 1985 Apr;16(4):211–215. [PubMed] [Google Scholar]
- JERNE N. K., NORDIN A. A. Plaque formation in agar by single antibody-producing cells. Science. 1963 Apr 26;140(3565):405–405. [PubMed] [Google Scholar]
- Jerne N. K., Henry C., Nordin A. A., Fuji H., Koros A. M., Lefkovits I. Plaque forming cells: methodology and theory. Transplant Rev. 1974;18:130–191. doi: 10.1111/j.1600-065x.1974.tb01588.x. [DOI] [PubMed] [Google Scholar]
- Kiyotaki M., Toguchi T., Abe T., Takeuchi T., Morimoto C., Homma M. In vitro anti-DNA antibody synthesis by peripheral blood lymphocytes in patients with systemic lupus erythematosus. Clin Immunol Immunopathol. 1981 Nov;21(2):237–246. doi: 10.1016/0090-1229(81)90212-9. [DOI] [PubMed] [Google Scholar]
- Loftager M. K., Koch C., Hellung-Larsen P., Andersen V. Conjugation of DNA to erythrocytes. J Immunol Methods. 1987 Aug 24;102(1):65–69. doi: 10.1016/s0022-1759(87)80010-8. [DOI] [PubMed] [Google Scholar]
- Loftager M. K., Koch C., Hellung-Larsen P., Andersen V. Immune hemolytic assay for identification of human anti-dsDNA antibodies with DNA-coated red blood cells as target cells. APMIS. 1988 Mar;96(3):194–198. doi: 10.1111/j.1699-0463.1988.tb05290.x. [DOI] [PubMed] [Google Scholar]
- Papalian M., Lafer E., Wong R., Stollar B. D. Reaction of systemic lupus erythematosus antinative DNA antibodies with native DNA fragments from 20 to 1,200 base pairs. J Clin Invest. 1980 Feb;65(2):469–477. doi: 10.1172/JCI109690. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raptis L., Menard H. A. Quantitation and characterization of plasma DNA in normals and patients with systemic lupus erythematosus. J Clin Invest. 1980 Dec;66(6):1391–1399. doi: 10.1172/JCI109992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Roder J. C., Bell D. A., Singhal S. K. Regulation of the autoimmune plaque-forming cell response to single-strand DNA (sDNA) in vitro. J Immunol. 1978 Jul;121(1):38–43. [PubMed] [Google Scholar]
- Sasaki T., Ishida S., Onodera S., Saito T., Furuyama T., Yoshinaga K. Passive hemagglutination and hemolysis tests for the detection of anti-DNA antibody. J Immunol Methods. 1978;22(3-4):327–337. doi: 10.1016/0022-1759(78)90040-6. [DOI] [PubMed] [Google Scholar]
- Swaak T., Smeenk R. Detection of anti-dsDNA as a diagnostic tool: a prospective study in 441 non-systemic lupus erythematosus patients with anti-dsDNA antibody (anti-dsDNA). Ann Rheum Dis. 1985 Apr;44(4):245–251. doi: 10.1136/ard.44.4.245. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tan E. M., Cohen A. S., Fries J. F., Masi A. T., McShane D. J., Rothfield N. F., Schaller J. G., Talal N., Winchester R. J. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum. 1982 Nov;25(11):1271–1277. doi: 10.1002/art.1780251101. [DOI] [PubMed] [Google Scholar]
- Valle R. R., Eaton R. B., Schnneider G., Schur P. H. Complement activation by antibodies to DNA in systemic lupus erythematosus measured by enzyme immunoassay. Clin Immunol Immunopathol. 1985 Mar;34(3):345–354. doi: 10.1016/0090-1229(85)90183-7. [DOI] [PubMed] [Google Scholar]
- Weill B. J., Heavey A. C., Kelley V. E., Schur P. H. Use of nuclear antigen-coated red cells in hemolytic plaque assays. J Immunol Methods. 1983 Feb 25;57(1-3):327–340. doi: 10.1016/0022-1759(83)90093-5. [DOI] [PubMed] [Google Scholar]
