The Reaction of the Lupus Erythematosus (L.E.) Cell Factor With Deoxyribonucleoprotein of the Cell Nucleus

Abstract

Previous reports have presented evidence that the lupus erythematosus (L.E.) cell factor reacts with deoxyribonucleoprotein of the cell nucleus, and that deoxyribonlucleic acid (DNA) is essential for the reactioni (1,2). More recently, a number of otlher serum factors lhave been identified in systemic lupus erythematosus (SLE) whiclh react with other conistituenits of the nucleus.These include isolated DNA, purified histone and a material extractable from the lnucleus witlh isotonic buffers wlhich is neither nucleic acid nor hiistonie (3,4 Because fractionation of the cell nucleus is still in a primitive stage anid because many of the nmethods employed in this study lhave not been extensively used anid verified, the experimenital dletails will be giveni together with the results. MATERIALS AND GENERAL METHODSSera were collected and stored under sterile conditionis at 4°C., usually with 1: 10,000 concenitrations of merthiolate.Nuclei were obtained from calf thymus glands at 4°C. by the method of Mirsky and Pollister (15). Glands were minced with scissors, homogenized in a solution of 0.25 M sucrose and 0.003 M CaCl, in a Waring blendor at a speed which did not disrupt the nuclei, filtered through gauze and flannel and the nuclei separated by centrifugation. The resulting nuclei were intact microscopically and had minimal cytoplas'mic contamination.For storage, nuclei were lyophilized.Nucleoprotein was prepared from unlyophilized nuclei by extraction overnight at 40 C. with 1 N NaCl (16). The resulting viscous suspension was centrifuged at 78,000 x G for 90 minutes in the Spinco "L" ultracentrifuge and the clear viscous supernate solution of the nucleoprotein stored at 4°C. Such solutions remained stable for at least 30 days under these conditions. Nucleoprotein was obtained for experimental use by diluting the stock solution with six volumes of cold distilled water, thus bringing it to physiologic salt concentration. Silvery nucleoprotein strands precipitated. These strands were separated by centrifugation and homogenized in a Teflon tissue grinder in isotonic saline. Equal volumes of the homogenate were then removed, spun, and pellets of solid, particulate nucleoprotein used for absorption. In general, nucleoprotein pellets containing 1 to 1.5 mg. DNA were used to absorb 0.5 ml. of serum, although pellets half that size are adequate to absorb completely 0.5 ml. of active serum if an incubation time of 45 minutes at 370 C. were allowed.Human white cell nuclei were prepared from leukocytes from patients with granulocytic and monocytic leukemia. The above methods were used except that the cells were disrupted in a Teflon tissue grinder in a solution of 0.25 2059

Role of igg and igm antinuclear antibodies in formation of lupus erythematosus cells and extracellular material

“…Whether a difference in ANA specificity may explain the lack of forma- (32,34). Furthermore, the LE factor can actually be eluted from DNP-LE factor complexes (33). T h e IgM ANA preparations used i n our study produced the homogeneous nuclear staining pattern characteristically produced by anti-DNP (37) although a speckled pattern seemed to be superimposed.…”

A new autoantibody in patients with rheumatoid arthritis: characterization of the anti–hat‐1 antibody system

“…In addition, the study included 10 patients with Sjogren's syndrome with sicca complex alone and 15 RA patients with Sjogren's syndrome diagnosed according to previously defined criteria (3), 33 patients with SLE who fulfilled at least 4 of the ARA 1982 revised criteria for the classification of SLE (4), 15 patients with PSS diagnosed in accordance with the criteria of Medsger and Masi (S), 15 patients with PM diagnosed in accordance with the criteria of Bohan and Peter (6), and 12 patients with mixed connective tissue disease diagnosed on the basis of previously described clinical signs and symptoms with associated high titers of antibody to nuclear RNP (nRNP) (7). Seven patients fulfilled criteria for 2 or more rheumatic diseases (1 patient SLE, PSS, and PM; 2 patients SLE and PSS; 2 patients SLE and RA; and 2 patients PSS and PM).…”

Differing accessibility in chromatin of the antigenic sites of regions 1-58 and 63-125 of histone H2B.

“…Nuclei were prepared from calf thymus by the procedure of Holman and Deicher (14) and incubated with micrococcal nuclease (0.3 U 110 ngl/A.U of chromatin, with 1 mM CaC12) for 1 min at 37°C . EDTA was added to a final concentration of 10 mM and the reaction mixture was chilled on ice.…”