Autoantibody to th ribonucleoprotein (nucleolar 7–2 rna protein particle) in patients with systemic sclerosis

Abstract

We studied sera of 371 consecutive new patients with systemic sclerosis (SSc; scleroderma) who were first evaluated during 1984-1988. All sera were tested for antinuclear antibodies by immunofluorescence staining using HEp-2 cells as substrate. We excluded 219 sera showing dark nucleoli and screened for antibodies to Th in the remaining 152 sera by immunoprecipitation of a 32P-labeled HeLa cell extract. Fifteen (4.0%) of 371 sera were anti-Th+. Anti-Th antibodies were present in 14 (8.4%) of 167 SSc patients with limited cutaneous involvement, in 1 of 167 with diffuse cutaneous involvement, and in 0 of 37 with SSc overlap syndrome. Among 244 controls with other connective tissue diseases, antiTh was detected in only 3 patients, all having primary Raynaud's phenomenon of <2 years duration. In the subgroup with SSc with limited cutaneous involvement, the 14 anti-Th+ patients had a significantly greater frequency of puffy lingers, small bowel involvement, and hypothyroidism, and a significantly lower frequency of arthralgia andor arthritis. Their cumulative survival rate from the time of onset of symptoms was lower than that for anti-Th-patients (78% versus 91% at 10 years), primarily due to 3 deaths from pulmonary arterial hypertension (2 from primary pulmonary hy-

A comparison between anti‐Th/To– and anticentromere antibody–positive systemic sclerosis patients with limited cutaneous involvement

“…All sera were diluted 1:40 in 10 mM phosphate buffered saline, pH 7.3, and tested for ACA by indirect immunofluorescence (IIF) performed on HEp-2 substrate (Diasorin, Stillwater, MN). Specimens exhibiting a nucleolar pattern on IIF were evaluated for anti-Th/To by RNA immunoprecipitation as previously described (4). Briefly, a 20-l serum sample was bound overnight at 4°C to 2 mg protein A-Sepharose CL-4B beads (Amersham Biosciences, Piscataway, NJ), washed 3 times with immunoprecipitation buffer (10 mM Tris HCl, pH 8.0, 500 mM NaCl, 0.1% Igepal), and incubated for 2 hours at 4°C with unlabeled extract from ϳ6 ϫ 10 6 rapidly dividing K562 cells.…”

“…In 1990, investigators in our group compared 14 anti-Th/To-positive lcSSc patients with a large group of anti-Th/To-negative lcSSc patients and showed that the patients with anti-Th/To antibody more often had puffy fingers, small bowel involvement, hypothyroidism, and poor survival (4). Two limitations of this and other prior reports on anti-Th/To antibody are the small number of patients evaluated and the relatively brief period of patient followup.…”

Clinical and Prognostic Associations Based on Serum Antinuclear Antibodies in Japanese Patients with Systemic Sclerosis

“…We performed indirect immunofluorescence tests using commercially prepared slides of monolayer HEp-2 cells (MBL, Nagano, Japan) as substrate (lo), double-immunodiffusion tests using calf thymus extracts as antigen source ( 3 3 , and immunoprecipitation assays using 32P-orthophosphate-labeled (15) (10). Anti-top0 I was considered positive if the serum antibody produced a precipitin line identical to that produced by the reference serum on double immunodiffusion, or if it immunoprecipitated a protein with a molecular weight of -100 kd from "S-methionine-labeled cell extract, inhibited top0 I enzymatic activity in vitro using commercially prepared calf thymus top0 I (Takara, Kyoto, Japan) according to the method of Goto et a1 (37), and reacted with any top0 I fusion protein including B cell epitope(s) on immunoblots, as described elsewhere (35).…”

Autoantibody to U3 Nucleolar Ribonucleoprotein (Fibrillarin) In Patients with Systemic Sclerosis

“…A serum sample was obtained from each patient at the time of the first visit. The patients were classified as having diffuse or limited cutaneous involvement, as described previously (9). One hundred ninety-one had diffuse cutaneous SSc, 174 had limited cutaneous SSc, and 51 were not classified.…”

“…A complete history, physical examination, and laboratory evaluation were obtained on each patient as previously described (9). The criteria used for organ system involvement were those described previously (9), but with several exceptions.…”