Succinate metabolism of human peripheral lymphocytes in immunoglobulin synthesis
1987 Volume 33 Issue 4 Pages 657-666
Details
1987 Volume 33 Issue 4 Pages 657-666
Human peripheral lymphocytes were cultured with succinatc 1, 4-14C, which is a TCA cycle intermediated metabolic substance, in the stimulation with pokeweed mitogen (PWM). It was recognized that 14C was taken into the lymphocytes, utilized for cell metabolism and finally synthesized into immunoglobulins.
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, 4-14C for 1, 3, 5 or 7 days. PWM was added as mitogen. Immunobeases were used for detecting immunoglobulins.
The results were as follows:
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.
2) The intracellular 14C 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.
3) 14C 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 14C in the macromolecular substances was 0.4% of that initially added.
4) Immunoglobulins (IgG, A and M) containing 14C were detected in the supernatant after 7 days culture with PWM. The maximum level of 14C was 5.6% of that detected in the macromolecular substances in the supernatant. The order of 14C content was B<UNF<T+B fraction (p<0.05).
