McCauliffe D P, Yin H, Wang L X, Lucas L
Department of Dermatology, University of North Carolina at Chapel Hill 27599.
J Rheumatol. 1994 Jun;21(6):1073-80.
To determine the reactivity of recombinant 52 and 60 kDa Ro(SSA) (Ro) proteins with sera from 3 subsets of patients with Ro autoantibody associated disease.
Complementary DNA (cDNA) clones that encode the human 52 and 60 kDa Ro autoantigens were isolated by the polymerase chain reaction and utilized to express recombinant glutathione S-transferase (GST) fusion proteins. Double immunodiffusion (ID) defined Ro positive autoimmune sera from 12 patients with neonatal lupus erythematosus (NLE), 16 with subacute cutaneous lupus erythematosus (SCLE) and 12 with primary Sjögren's syndrome (SS) were tested by enzyme linked immunosorbent assay (ELISA) against the recombinant 52 and 60 kDa Ro fusion proteins.
Seventy-five percent of NLE, 56% of SCLE and 83% of SS sera reacted with the 52 kDa fusion protein. Seventy-five percent of NLE, 63% of SCLE and 83% of SS sera reacted with the 60 kDa fusion protein. Seventeen percent of NLE sera, 25% of SCLE sera and 8% of SS sera were nonreactive to both full length fusion proteins. Eight (57%) of 14 ID defined Ro negative NLE, SCLE and SS sera were reactive with both Ro fusion proteins by ELISA: ELISA studies with recombinant 52 and 60 kDa Ro protein fragments revealed at least 2 major epitopes on each Ro protein. A fragment of the 52 kDa Ro protein that contains a putative leucine zipper motif reacted with 100% of ID defined Ro positive SS sera.
Our data demonstrate that the ID assay and the recombinant Ro ELISA together are more sensitive in detecting Ro antibodies than either assay alone, and that multiple epitopes are present on both Ro proteins.
确定重组52 kDa和60 kDa Ro(SSA)（Ro）蛋白与Ro自身抗体相关疾病3个亚组患者血清的反应性。
通过聚合酶链反应分离编码人52 kDa和60 kDa Ro自身抗原的互补DNA（cDNA）克隆，并用于表达重组谷胱甘肽S-转移酶（GST）融合蛋白。通过酶联免疫吸附测定（ELISA），用重组52 kDa和60 kDa Ro融合蛋白检测12例新生儿红斑狼疮（NLE）、16例亚急性皮肤型红斑狼疮（SCLE）和12例原发性干燥综合征（SS）患者经双向免疫扩散（ID）确定的Ro阳性自身免疫血清。
75%的NLE、56%的SCLE和83%的SS血清与52 kDa融合蛋白反应。75%的NLE、63%的SCLE和83%的SS血清与60 kDa融合蛋白反应。17%的NLE血清、25%的SCLE血清和8%的SS血清对两种全长融合蛋白均无反应。14例经ID确定为Ro阴性的NLE、SCLE和SS血清中有8例（57%）通过ELISA与两种Ro融合蛋白反应：对重组52 kDa和60 kDa Ro蛋白片段的ELISA研究显示，每种Ro蛋白上至少有2个主要表位。含有假定亮氨酸拉链基序的52 kDa Ro蛋白片段与100%经ID确定的Ro阳性SS血清反应。
我们的数据表明，ID试验和重组Ro ELISA联合检测Ro抗体比单独使用任何一种试验更敏感，并且两种Ro蛋白上都存在多个表位。
