Decreased immunoglobulin production by a human lymphoid cell line following melphalan treatment.

The effect of melphalan on immunoglobulin G (IgG) production by a human lymphoblastoid cell line (BF) was studied. The amount of secreted IgG and the percentage of cells containing cytoplasmic IgG were measured by immunoassay and cytofluorometry, respectively. Dose-response studied indicated that melphalan concentrations of 2 X 10(-8) M had no effect, while concentrations of 8 X 10(-7) M were totally toxic, after 72-hr exposures to the drug. Statistically significant, persistent, alterations in both synthesis and secretion of IgG by BF cells were observed following treatment for 72 hr with 4 X 10(-7) M melphalan, and there was an increase in population-doubling time from 24 to 72 hr in these drug-treated cells. The percentage of IgG-containing cells in melphalan-treated cultures was significantly decreased as compared to control cultures. IgG secretion was also decreased in these cultures, and the variation in IgG secretion as a function of cellular growth was significantly altered following melphalan treatment. Decreased IgG production following melphalan treatment may be related to altered cell cycle kinetics. Based on immunological analysis, there was no evident alteration in the IgG secreted by melphalan-treated cells, nor did melphalan treatment produce a cellular population lacking IgG entirely.

Separation of transfer ribonucleic acids on polystyrene anion exchangers.

The transfer RNA separation by chromatography on strong-base polystyrene exchange materials is examined and compared with the widely used reversed-phase chromatography. Results indicate important differences in some transfer RNA (tRNA) elution patterns by the anion-exchange chromatography, as compared with the reversed-phase chromatography. Transfer RNAs containing hydrophobic groups are adsorbed more strongly. The anion exchanger has twice the number of theoretical plates. Single peaks of tRNA2Glu and tRNA1Phe obtained from the reversed-phase column give multiple peaks only polystyrene anion-exchange chromatography. All six leucine tRNAs (Escherichia coli) and differences in tRNA populations synthesized during early and late stages of the dividing lymphocytes from normal human blood can be characterized by the anion-exchange chromatography. Different separation profiles are obtained by two separation systems for tyrosine tRNAs from mouse liver and mouse-plasma-cell tumor. The results indicate that, in contrast to the reversed-phase chromatography, strong-base-polystyrene anion-exchange chromatography is capable of separating tRNAs with minor structlral differences.

Isoaccepting transfer RNA's of L-M cells in culture and after tumor induction in C3H mice.

Co-chromatography in a reversed-phase column was performed for 16 aminoacyl-tRNA's prepared from L-M cells grown in serum-free suspension culture and from tumors induced in irradiated C(3)H mice by subcutaneous injection of L-M cells. The results showed that between the two sources there were (1) marked differences in aspartyl-, histidyl-, phenylalanyl-, and tyrosyl-tRNA's; (2) significant quantitative differences in isoaccepting species of alanyl-, isoleucyl-, seryl-, and threonyl-tRNA's; and (3) similar to minimally different patterns of arginyl-, methionyl-, prolyl-, tryptophanyl-, valyl-, glycyl-, leucyl-, and lysyl-tRNA's. The differences were evident when synthetases prepared either from L-M cells or from the tumors were used. When the L-M tumors were brought back into in vitro culture, their tRNA patterns were like those of L-M cells. Addition of serum to the culture medium caused the L-M cells to show very minute, but detectable, amounts of the isoaccepting tRNA's found in the tumors. The cellular mechanisms which may be related to the changes of tRNA patterns in the L-M cells are discussed.