A Cytochemical Study of the L.E. Bodies of Systemic Lupus Erythematosus

Godman, Gabriel C.; Deitch, Arline D.

doi:10.1084/jem.106.4.575

Cited by 30 publications

(11 citation statements)

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“…The requirement of both DNA and histone is compatible with the results of current histochemical studies of the L.E. cell phenomenon (25).…”

Section: Discussionsupporting

confidence: 89%

“…The above methods were used except that the cells were disrupted in a Teflon tissue grinder in a solution of 0. 25 Two 100 per cent units of complemenit were used and visual endpoinits recorded with appropriate serum and antigein controls. When materials containin1g DNase were used for complement fixation, sodlium citrate was added in excess of the magnesiumii conceintrationi and all reagents employed were free of both calcium and magnesium.…”

Section: Materials and General Methodsmentioning

confidence: 99%

See 1 more Smart Citation

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

Holman¹,

Deicher²

1959

J. Clin. Invest.

135

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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

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“…The requirement of both DNA and histone is compatible with the results of current histochemical studies of the L.E. cell phenomenon (25).…”

Section: Discussionsupporting

confidence: 89%

Section: Materials and General Methodsmentioning

confidence: 99%

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

Holman¹,

Deicher²

1959

J. Clin. Invest.

135

View full text Add to dashboard Cite

show abstract

“…It has been reported that fibroblasts in vivo incorporate DNA by phagocytosis (11), as do white blood cells in the lupus phenomena (12). Determina?…”

Section: Cleus the Cells Were Fixed In A Solu? Tion Of Acetic Acid Amentioning

confidence: 99%