Inhibition of Adenovirus DNA Synthesis In Vitro by Sera from Patients with Systemic Lupus Erythematosus

Abstract

Sera containing antinuclear antibodies from patients with systemic lupus erythematosus (SLE) and related disorders were tested for their effect on the synthesis of adenovirus (Ad) DNA in an in vitro replication system. After being heated at 60°C for 1 h, some sera from patients with SLE inhibited Ad DNA synthesis by 60 to 100%6. Antibodies to double-stranded DNA were present in 15 of the 16 inhibitory sera, and inhibitory activity copurified with anti-doublestranded DNA in the immunoglobulin G fraction. These SLE sera did not inhibit the DNA polymerases a, (, y and had no antibody to the 72,000-dalton DNAbinding protein necessary for Ad DNA synthesis. The presence of antibodies to single-stranded DNA and a variety of saline-extractable antigens (Sm, Ha, nRNP, and rRNP) did not correlate with SLE serum inhibitory activity. Methods previously developed for studying the individual steps in Ad DNA replication were used to determine the site of inhibition by the SLE sera that contained antibody to double-stranded DNA. Concentrations of the SLE inhibitor that decreased the elongation of Ad DNA by greater than 85% had no effect on either the initiation of Ad DNA synthesis or the polymerization of the first 26 deoxyribonucleotides.Sera from patients with systemic lupus erythematosus (SLE) and other connective tissue diseases contain antibodies to a wide variety of nuclear and cytoplasmic macromolecules, including single-stranded (ss) and double-stranded (ds) DNA, RNA, nucleohistones, and ribonucleoprotein (RNP) particles (29). These antibodies may distinguish between different disease entities or prognostic subclasses and, in the case of antibodies to dsDNA, provide a means for monitoring the level of SLE disease activity (23,29,35). Although the antigens recognized by these antibodies play critical roles in intracellular processes, the pathogenicity of antibodies to these macromolecules is unknown.To evaluate the possibility that the various antinuclear antibodies may alter cellular functions, we studied the effects of such antibodies on the replication of adenovirus (Ad) DNA in vitro by subcellular soluble fractions. In this model system, Ad DNA synthesis is initiated on template dsDNA which contains a 55,000-dalton

Inhibition of adenovirus DNA replication in vitro by autoimmune sera

“…2). A similar observation was made by Horwitz et al [17]. This may be caused by the presence of other weak inhibitors of DNA synthesis which are not inactivated by a 60 C treatment.…”

“…Possible interference with othcr iiitracellular processes like DNA replication may have escaped detection so far due to a lack of suitable iri 1ifr.o assay systems. As discussed earlier by Horwitz et al [17], the adenovirus DNA replication system might be usefill for further investigations of the nature of antibodies directed against DNA replication components.…”

“…Horwitz et al [17] found that Ad DNA replication in crude extracts was inhibited by sera from patients with systemic lupus erythematosus. The inhibition by all but one of these sera was related to the presence of antibodies to doublestranded (ds) DNA.…”

Effect of Enhanced Prostacyclin Synthesis by Adenovirus‐Mediated Transfer on Lipopolysaccharide Stimulation in Neuron‐Glia Cultures

“…1A). This may be due to the presence of serum proteins, such as serum albumin, which attenuate adenoviral infectivity by nonspecific binding to adenovirus 23,24. We performed double‐immunostaining of GFP with various cell markers in serum‐containing Ad‐GFP‐infected cultures.…”