Heterogeneity of RNP and Sm autoantigens in relation to the cell sources and the activated state of the cells.

The extracts of rabbit thymus (RTE), HeLa cells, human histiocytic lymphoma cell line U-937, human promyelocytic cell line HL-60, Ehrlich ascites tumor cells (EACs), and peripheral white blood cells (WBCs) were tested for their composition, molecular weight, and amount of Sm and RNP autoantigens on immunoblotting. The molecular weight of the so-called 68 kDa U1 RNP antigen, which is associated with mixed connective tissue disease (MCTD), was 64.5 kDa in RTE, 62.5 kDa in HeLa cells and HL-60 cells, and 59 kDa in WBCs. Surprisingly, in both WBCs and U-937 cells, the main protein band bearing the 68 kDa U1 RNP antigenic determinants was 30.5 kDa in molecular weight, which was confirmed using antibodies purified by affinity chromatography. After stimulation with phytohemagglutinin (PHA), there was in the human lymphocytes a diminished amount of the 30.5 kDa protein and simultaneously an increased synthesis of several proteins of higher molecular weight, especially the 57 kDa protein bearing the 68 kDa antigenic determinants. The concentrations of the A, B/B', C, D, and E proteins also increased with PHA stimulation. Our results indicate that the expression of Sm and RNP autoantigens may depend on the cell source as well as the activated state of cells. These differences should be taken into consideration in the detection of anti-RNP and anti-Sm antibodies by immunoblotting.