Conjugates or dsDNA Linked to Human Gammaglobulin Inhibit Anti-dsDNA Antibodies In Vitro

Abstract

Previous studies have shown that both nucleosides and oligonucleotides linked to isologous gammaglobulin suppress anti-nucleic acid antibody production both in vivo and in vitro. The aim of this study was to determine whether one can make a DNA-human gammaglobulin (HGG) conjugate which can inhibit anti-double stranded DNA (dsDNA) antibodies obtained from a heterogeneous population of systemic lupus eruthematosus (SLE) sera. To do so, we constructed conjugates of sonicated dsDNA fragments of 100-400 base pairs covalently linked to HGG with varying degrees of substitution of DNA:HGG. An ELISA inhibition assay was used to determine which conjugate best inhibits the binding of anti-dsDNA antibodies. Conjugate 2, prepared with monomeric HGG (150 kD) with a high degree of substitution (3.72 DNA:HGG) inhibited the binding of anti-dsDNA antibodies from 27 of 31 SLE sera. In addition, this conjugate inhibited the spontaneous formation of anti-dsDNA in vitro by cultured lymphoid cells from selected SLE patients. Together, this data suggests that a 'generic' tolerogen may provide an antigen specific therapy for SLE.

Keywords

Systemic lupus erythematosus Anti-dsDNA antibody dsDNA-human gammaglobulin conjugates

Get full access to this article

View all access options for this article.

References

Tan EM Autoantibodies to nuclear antigens: their immunobiology and medicine . Adv Immunol 1982; 33: 167-240.

Schwartz RS , Stollar BD Origin of anti-DNA autoantibodies. J Clin Invest 1985; 75: 321-7.

Emlen W. , Pisetsky DS , Taylor RP Antibodies to DNA: a perspective. Arthritis Rheum 1986; 29: 1417-26.

Rothfield NE , Stollar BD The relation of immunoglobulin class, pattern of antinuclear antibody and complement-fixing antibodies to DNA in sera from patients with systemic lupus erythematosus. J Clin Invest 1967; 46: 1785-94.

Ter Borg EJ , Horst G. , Hummel EJ , Limburg PC , Kallenberg Cgm. Predictive value of rises in anti-double stranded DNA antibody levels for disease exacerbation in systemic lupus erythematosus: a long-term prospective study. Arthritis Rheum 1990; 33: 634-43.

Borel Y. , Lewis RM , Stollar BD Prevention of murine lupus nephritis by carrier dependent induction of immunologic tolerance toenatured DNA. Science 1973; 182: 76-8.

Borel H. , Sasaki T. , Stollar DB , Borel Y. Conjugation of DNA fragments to protein carriers by glutareldehyde: immunogenicity of oligonucleotide-hemocyanin conjugates. J Immunol Methods 1984; 67:289-302.

Borel H. , Bastian D. , Cooper B. A possible new therapy of systemic lupus erythematosus. In: Autoimmunity. Schwartz RS , Rose NR (eds). New York: Academy of Science, 1986: 296-306.

Borel H. , Borel Y. Oligonucleotide linked to human gammaglobulin specifically diminishes anti-DNA antibody formation in cultured lymphoid cells from patients with systemic lupus erythematosus. J Clin Invest 1988; 82: 1901-7.

10.

Lowry OH , Rosenbrough NJ , Farr AL , Randall RJ Protein measurement with the folin phenol reagent. J Biol Chem 1951; 193: 265-75.

11.

Smith PK , Krohn RI , Hermanson GT et al. Measurement of protein using bicinchoninic acid. Anal Biochem 1985; 150: 76-85.

12.

Tan EM , Cohen AS , Fries J. The 1982 revised criteria for the classification of systemic lupus erythematosus . Arthritis Rheum 1982; 25: 1271-7.

13.

Tan EM , Fritzler MJ , McDougal JS Reference sera for antinuclear antibodies. 1. Antibodies to native DNA, SM, nuclear RNP and SSB/ La. Arthritis Rheum 1982 ; 25: 1003-6.

14.

Koffler D. , Shur PH , Kunkel HG Immunologic studies concerning the nephritis of systemic lupus erythematosus . J Exp Med 1967; 126: 607-23.

15.

Lambert PH , Dixon FJ Pathogenesis of glomerulonephritis of the NZB/W mice. J Exp Med 1968; 127: 507-21.

16.

Winfield JB , Faiferman I. , Koffler D. Avidity af anti-DNA antibodies in serum and IgG glomerular eluates from patients with systemic lupus erythematosus: association of high avidity antinative DNA antibody with glomerulonephritis. J Clin Invest 1977; 59: 90-6.

17.

Papalian M. , Lafer E. , Wong R. , Stollar BD Reaction of systemic lupus erythematosus antinative DNA with native DNA fragments from 20 to 1200 base pairs. J Clin Invest 1980; 65: 469-77.

18.

Fournie GJ Circulating DNA in lupus nephritis. Kidney Int 1988; 33: 487-97.

19.

Li JZ , Steinman CR Plasma DNA in systemic lupus erythematosus. Arthritis Rheum 1989; 32: 726-33.

20.

Rumore PM , Steinman CR Endogenous circulating DNA in systemic lupus erythematosus. Occurrence as multimeric complexes bound to histone. J Clin Invest 1990; 86: 69-74.

21.

Gaedicke G. , Teller WM , Kohne E. et al. IgG therapy in systemic lupus erythematosus. Two case reports. Blut 1984; 48: 387-90.

22.

Corvetta A. , Della Bitta R. , Gabrielli A. et al. Use of high dose intravenous immunoglobulin in systemic lupus erythematosus: report of three cases. Clin Exp Rheum 1989; 7: 295-9.

23.

Akashi K. , Nagasawa K. , Mayumi T. et al. Successful treatment of refractory systemic lupus erythematosus with intravenous immunoglobulins. J Rhetimatol 1990; 17: 375-9.

24.

Gajdos PH , Outin H. , Elkharrat D. et al. High-dose intravenous gammaglobulin for myasthenia gravis. Lancet 1984; i: 406-7.

25.

Imbach P. , Wagner H. , Berchtold W. et al. Intravenous immunoglobulin versus oral corticosteroids in acute immune thrombocytopenic purpura in childhood. Lancet 1985; 2: 464-8.

26.

Carreras LO , Perez GN , Vega HR et al. Lupus anticoagulant and recurrent fetal loss: successful treatment with gammaglobulin. Lancet 1988; ii: 393-4.

27.

Said PB , Martinuzzo ME , Carreras LO Neutralization of lupus anticoagulant activity by human immunoglobulin in vitro. Nouv Rev Fr Hematol 1992; 34: 37-42.

28.

Sultan Y. , Kazatchkine MD , Maisonneuve P. et al. Anti-idiotypic suppression of autoantibodies to factor VIII (antihaemophilic factor) by high dose intravenous gammaglobulin. Lancet 1984; ii: 765-8.