Role of serum in inhibition of cultured lymphocytes by lysophosphatidylcholine.
Serum was heated at various temperatures to inactivate components which might be involved in the regulation of lysophosphatidylcholine (lysoPC) levels in rabbit lymph-node cell cultures. Cells cultured in medium containing serum preheated for 20 min at 66 degrees C ("66 degrees C-serum") were inhibited much more by exogenous lysoPC (5 microM) than were cells cultured in medium containing control serum ("38 degrees C-serum"). This was observed over a 20 h culture period as a slow increase in inhibition of cell labelling with [3H] uridine, which reflected cytotoxic cell damage. Heating serum at 66 degrees C caused (i) conversion of monomeric albumin to highly polymeric forms which were deficient in lysoPC-binding activity, (ii) transfer of lysoPC from albumin to lipoproteins, predominantly high density lipoproteins, and (iii) inhibition of two lysoPC metabolizing activities (which were detected only at low levels in control serum). Addition of albumin to cultures containing 66 degrees C-serum decreased the toxicity of lysoPC to the same extent as did the addition of control serum with an equivalent albumin content. Thus, albumin was the major heat-labile factor protecting cells against lysoPC. However, cell inhibition by lysoPC was dependent on the sequence of heating serum and lysoPC addition. Inhibition was small when lysoPC was added before heating the serum. This could not be explained by a detectable difference in the binding of lysoPC to serum components. Furthermore, although radioactive labelling of cells with [14C] lysoPC was increased in 66 degrees C-serum, this did not correlate with cell inhibition. Increased labelling with [14C] lysoPC occurred several hours before significant cell inhibition was evident and was not affected by the sequence of heating and lysoPC addition. Since preincubation of lysoPC with 66 degrees C-serum increased the inhibition, it is suggested that the heated serum lysoPC generates another factor which is responsible for the cytotoxic effects observed.
D000818:Animals; D001769:Blood; D002478:Cells, Cultured; D003470:Culture Media; D007700:Kinetics; D008214:Lymphocytes / Q000187:drug effects* / Q000502:physiology*; D008244:Lysophosphatidylcholines / Q000494:pharmacology*; D010741:Phospholipases A / Q000494:pharmacology; D012313:RNA / Q000096:biosynthesis; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013696:Temperature
D003470:Culture Media; D008244:Lysophosphatidylcholines; D012710:Serum Albumin, Bovine; D012313:RNA; D010741:Phospholipases A