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Antibody synthesis in synchronized mouse spleen cells during the secondary response to sheep erythrocytes in vitro
Immunoglobulin synthesis by mouse spleen cells during the secondary response to sheep erythrocytes in vitro was examined in cells synchronized by methotrexate and adenosine treatment. Antibody synthesis by the synchronized cells was discontinuous through the cell cycle and highest in the early S phase. Previous results from this laboratory indicated that antibody secretion, as measured by plaque production to sheep erythrocytes, was also greatest in early S, and taken together these results suggest little difference in the expression of antibody synthesis and secretion through the cell cycle. Actinomycin D was employed in several experiments in an attempt to determine if the variation in antibody synthesis was regulated by transcription of messenger RNA for immunoglobulin. Lower actinomycin concentrations (0.05 mug/ml) reportedly permitting messenger, but not ribosomal, RNA synthesis as well as higher concentrations (1 mug/ml) rapidly reduced immunoglobulin and total protein synthesis. These results indicated that actinomycin was not suitable for determining the stability of messenger RNA in mouse spleen cells. Therefore, the reasons for fluctuation in antibody synthesis during the cell cycle could not be determined, but several possibilities are suggested.
Immunoglobulin synthesis by mouse spleen cells during the secondary response to sheep erythrocytes in vitro was examined in cells synchronized by methotrexate and adenosine treatment. Antibody synthesis by the synchronized cells was discontinuous through the cell cycle and highest in the early S phase. Previous results from this laboratory indicated that antibody secretion, as measured by plaque production to sheep erythrocytes, was also greatest in early S, and taken together these results suggest little difference in the expression of antibody synthesis and secretion through the cell cycle. Actinomycin D was employed in several experiments in an attempt to determine if the variation in antibody synthesis was regulated by transcription of messenger RNA for immunoglobulin. Lower actinomycin concentrations (0.05 mug/ml) reportedly permitting messenger, but not ribosomal, RNA synthesis as well as higher concentrations (1 mug/ml) rapidly reduced immunoglobulin and total protein synthesis. These results indicated that actinomycin was not suitable for determining the stability of messenger RNA in mouse spleen cells. Therefore, the reasons for fluctuation in antibody synthesis during the cell cycle could not be determined, but several possibilities are suggested.