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The Journal of Immunology, Vol 139, Issue 2 393-399, Copyright © 1987 by American Association of Immunologists

ARTICLES

Distribution and specificity of nucleotide-reactive autoantibodies in human SLE

TW Munns, SK Freeman, MK Liszewski and JL Kaine

An enzyme-linked immunosorbent assay (ELISA) was utilized to characterize nucleotide-reactive antibodies present in the sera of 67 human subjects: 27 active SLE, 20 inactive SLE, and 20 asymptomatic controls. This assay consisted of measuring the quantity of antibodies retained by a panel of immobilized 5'-nucleotide-BSA conjugates (AMP-, GMP-, CMP-, UMP-, and TMP-BSA) together with ssDNA and dsDNA antigens. Although the relative distribution of antibodies binding to nucleotide- BSA antigens (i.e., anti-GMP greater than anti-AMP greater than or equal to anti-TMP greater than anti-UMP greater than or equal to anti- CMP antibodies) was independent of clinical status, the sera of active SLE patients possessed three- and five-fold higher concentrations of these antibodies relative to those present in inactive SLE and control subjects, respectively. Affinity purification of the most dominant of these antibody populations with DNA- and GMP-agarose adsorbents suggested that the majority of anti-GMP antibodies were monospecific with respect to the guanine base moiety. For example, antibodies retained by GMP-agarose reacted with GMP-BSA and ssDNA but not with other nucleotide-BSA or dsDNA antigens. However, ELISA competition- inhibition studies with affinity-purified anti-GMP antibodies indicated that although the guanine base represents an important determinant, guanine-enriched oligo- and polynucleotides were preferred substrates (i.e., guanine-dependent, oligonucleotide specificity). This was exemplified by the finding that a 500- and 50-fold molar excess of dGMP and d(G)4 were required to achieve the same degree of inhibition as that observed with d(G)8. Finally, and as evaluated by indirect immunofluorescence with fixed HEp-2 cells, affinity-purified anti-GMP antibodies reacted with antigens restricted to nucleolar organelles.




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