Succinate metabolism of human peripheral lymphocytes in immunoglobulin synthesis.
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Human peripheral lymphocytes were cultured with succinatc 1, 4-<SUP>14</SUP>C, which is a TCA cycle intermediated metabolic substance, in the stimulation with pokeweed mitogen (PWM). It was recognized that <SUP>14</SUP>C was taken into the lymphocytes, utilized for cell metabolism and finally synthesized into immunoglobulins.<BR>The lymphocytes were separated into T cell enriched and B cell enriched fractions by En-rosette formation method, and equal numbers of these fractionated cells were mixed for T+B fraction. Each fraction was cultured with 5μCi/m/ of succinatc 1, 4-<SUP>14</SUP>C for 1, 3, 5 or 7 days. PWM was added as mitogen. Immunobeases were used for detecting immunoglobulins.<BR>The results were as follows:<BR>1) 14C concentration in the unfractionated lymphocytes (UNF) kept at low level for 1-7 days when cultured without PWM. However stimulating with PWM increased tripled the level the first day, and then they decreased with day.<BR>2) The intracellular <SUP>14</SUP>C concentration cultured with PWM for 7 days was the following order of T, UNF < T+B, B fraction (p<0.05). These differences suggested the influence of B cell ratio against T cell.<BR>3) <SUP>14</SUP>C concentration in the macromolecular substances detected in the PWM-stimulated culture supernatant increased with time from the third day on (p<0.05). The maximum value of <SUP>14</SUP>C in the macromolecular substances was 0.4% of that initially added.<BR>4) Immunoglobulins (IgG, A and M) containing <SUP>14</SUP>C were detected in the supernatant after 7 days culture with PWM. The maximum level of <SUP>14</SUP>C was 5.6% of that detected in the macromolecular substances in the supernatant. The order of <SUP>14</SUP>C content was B<UNF<T+B fraction (p<0.05).
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