We have studied the autoantibodies in the serum of a patient with linear scleroderma that specifically recognize the nuclear envelope of cultured cells. These antibodies bind to conserved determinants of nuclear lamins, the predominant mammalian nuclear envelope proteins. Of the three mammalian nuclear lamin proteins (P70, P68, and P60), only P70 and P60 bind the autoantibodies. In addition, two proteins of the Drosophila embryonic nuclear matrix, P70 and P68, bind these autoantibodies. We have used nuclear matrices to isolate the autoantibodies from the patient's serum that react to the nuclear lamins. At least three different IgG heavy chains were found to be involved in this autoimmune response to nuclear lamins, indicating that this response is not due to the expansion of a single B-cell clone.A strong correlation exists between systemic rheumatic diseases and the presence of circulating autoantibodies to proteinnucleic acid complexes (1). The targets of these autoantibodies include nuclear ribonucleoprotein particles (2), nucleosomes (3), nucleoli (4), kinetochores (5), and others. The range of targets for particular disease syndromes are often limited; for example, systemic lupus erythematosis is often associated with autoantibodies to nuclear ribonucleoprotein particles (2), whereas most scleroderma CREST patients produce autoantibodies to centromeric regions of chromosomes (kinetochores; ref. 5). Surprisingly, little is known of the autoimmune responses or their targets on a molecular level.In an attempt to associate the immunofluorescence pattern of sera from patients with specific autoimmune diseases, we discovered a high-titer serum directed against the nuclear envelope in a patient with linear scleroderma. We designate this serum as LS-1. We have chosen here to characterize this autoimmune response more precisely by identifying the antigens recognized by this serum and by determining the nature and complexity of the antibody response. We have found that this autoimmune response is polyclonal and directed against two of the three predominant polypeptides of the nuclear envelope, known as nuclear lamins (6).Using immunofluorescence and immunotransfers of NaDodSO4 gels, we also have detected nuclear lamins in Drosophila melanogaster embryonic nuclear matrices. This demonstrates that nuclear lamins are present in invertebrates and that these autoantibodies from LS-1 serum must be directed against highly conserved determinants. at 26°C in Echalier's medium (7).Immunofluorescence Microscopy. CHO and Kc cells were grown on glass coverslips (12-mm diameter, Corning), fixed in 1% formaldehyde in phosphate-buffered saline (Pi/NaCl) for 5 min at 22°C, and made permeable to antibodies by washing in 0.1% Triton X-100 (Sigma) in Pi/NaCl. The cells were first exposed to LS-1 serum diluted 1:200 and subsequently to a rhodamine-conjugated goat anti-human IgG (Cappel Laboratories, Cochranville, PA). Cells were photographed with a Zeiss photomicroscope with a X 100 planapo objective.Immunohistochemical Microsc...

Exposure of normal human skin in vivo to ultraviolet irradiation at wavelengths shorter than 320 nanometers stimtulated an unscheduled DNA synthesis in all of the cell layers of the epidermis and in the upper dermnial fibrocytes. The skin of patients with xeroderma pigmentosum did not show this response. correlation of these findings with previous tissue culture studies suggests that the defect in repair of the damaged DNA in xeroderma cells occurs in vivo as well as in vitro.

Acute effects of ultraviolet radiation on the mitotic cycle and macromolecular synthesis were investigated on hairless mouse epidermis in vivo. Colcemid was used to arrest mitoses in metaphase and thus allow more accurate mitotic counts. The radioactive tracers, TdR-3H, ~ytidine-~H, and the amino acids, hi~tidine-~H and methi~nine-~H were used to examine DNA, RNA and protein synthesis, respectively. Using these techniques, we found that wavelengths shorter than 320 nm markedly inhibited mitosis, increased the basal cell turnover time and depressed DNA, RNA and protein synthesis within the first few hours postirradiation. By 24 hr, recovery and acceleration of these functions were in progress, reaching a peak by 48-72 hr and persisting though to a lesser degree for 7 days. This stage of acceleration was associated with epidermal hyperplasia and most likely represented post-injury cell renewal.. METHODS A N D MATERIALS Experimental animalsOne hundred and fourteen, 2 month old, random bred, female albino hairless mice were housed in metal cages and fed on unrestricted quantities of Wayne Laboratory Blox and water. Natural sunlight was excluded and artificial white light exposure was minimal except during observation and treatment. Light source A Hanovia air cooled hot quartz contact lamp which emitted 14.96 x lo5 ergs/cm2/ sec of U.V. energy shorter than 320 nm at a distance of 3.4 cm was used as the light source. The energy was measured with a Hanovia ultraviolet light meter (Model AV-97 1). 57 58 Mitotic studiesProcedure. The right flanks of twenty mice were exposed to 4.49 X los ergs/cm2 of the short U.V. energy using the hot quartz contact source at a distance of 3.4 cm. At intervals of 1,3,7,24, and 72 hr, two mice were injected intraperitoneally with 30 pg of colcemid and autopsied 1 hr later. In addition, two irradiated animals which did not receive the colcemid were autopsied at each post-u.v. time period to provide an instantaneous picture of mitotic conditions. All autopsies were performed at 4 : 00 p.m. to eliminate diurnal variations. Flap biopsies from the irradiated right flank and nonirradiated left flank skin were obtained at that time. The specimens were fixed in Bouin's solution, embedded in paraffin, sectioned at 4 p and stained with hematoxylin and eosin (H & E). The colcemid was used to arrest mitoses in metaphase to allow accurate mitotic counts[6, 71. Preliminary studies had established that I hr was the optimal interval between colcemid injection and biopsy for these counts in our system. Metaphase counts were made along 1 cm of interfollicular epidermis.Results. An average of 1696 2 198 basal cells per cm were present in non-irradiated and irradiated epidermis. None of the basal cell counts in the irradiated tissue varied from the control average by a single standard deviation.The effects of U.V. irradiation on the metaphase counts per 1 cm of tissue are illustrated in Table 1 and Fig. 1. As noted, there was a marked depression in mitoses within 1 hr post-irradiation which persisted for at l...

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This study was designed to chemically characterize the principal structural proteins of psoriatic scales. Cornified cells were obtained from 40 patients with psoriasis, 21 patients with other scaly diseases, and 13 normal individuals. Cells were washed with Tris-HCl buffer and incubated in 8 M urea containing 2-mercaptoethanol (pH 9.0) at 30 degrees C for 7 hr. Extracted proteins were subjected to SDS polyacrylamide gel electrophoresis and protein patterns from normal and diseased scales were compared. The 67,000 dalton constituent of normal cornified cells could not be identified in protein from psoriatic scale and instead, a pair of polypeptides of approximately 54,000 and 57,000 daltons appeared. These extra bands were not found in protein extractions from other skin diseases, uninvolved skin of psoriasis patients, or normal skin. In order to analyze further normal and psoriatic scale proteins, the immunoreaction of rabbit antisera to human 67,000 dalton polypeptide with extracted psoriasis protein and with frozen biopsy sections, was studied using immunoprecipitation tests and indirect immunofluorescence microscopy. Both techniques demonstrated the existence of the 67,000 dalton protein in psoriasis, but as a minor component. These results indicate that additional unique urea mercaptoethanol soluble proteins are formed in psoriatic lesions, and this unusual protein synthesis may reflect the morphological changes in this disease.