The responses of two low-molecular-weight stress proteins, alpha B crystallin and HSP28, to various types of stress were determined quantitatively by specific immunoassays in a human glioblastoma cell line (U118 MG). Levels of alpha B crystallin (2-4 ng/mg protein) and HSP28 (1-1.5 micrograms/mg protein) in the soluble fraction from cells that had been cultured at 37 degrees C increased about 100-fold and 3-fold, respectively, within 24 h after heat treatment for 15 min at 45 degrees C, with a temporary decrease, due to redistribution to the insoluble fraction, during the heat treatment. Exposure of cells to arsenite (NaAsO2, 100 microM for 1 h) also induced the two proteins with a time course similar to that observed after heat stress, but without a decrease during the stress period. L-Azetidine-2-carboxylate (5 mM for 5 h) was also effective in inducing the two proteins, but to a lesser extent. Other chemicals, including CdCl2, ZnCl2, AlCl3, ethanol, caffeine, nicotine, NaN3, dibutyryl 3',5'-cyclic AMP, forskolin, and a phorbol ester, did not induce the two proteins. Expression of alpha B crystallin and HSP28 mRNAs in cells was enhanced after heat stress and after exposure to arsenite. When cells were challenged with heat stress in the presence of arsenite, the effect on the induction of the two proteins was synergistic. Ethanol (1-2%) enhanced the responses to heat stress or arsenite stress.(ABSTRACT TRUNCATED AT 250 WORDS)