METABOLIC CHANGES IN ACTIVATED T-CELLS - AN NMR-STUDY OF HUMAN PERIPHERAL-BLOOD LYMPHOCYTES
METABOLIC CHANGES IN ACTIVATED T-CELLS - AN NMR-STUDY OF HUMAN PERIPHERAL-BLOOD LYMPHOCYTES
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DOI：
10.1002/mrm.1910290307
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发表时间：
1993-03-01
影响因子：
3.3
通讯作者：
DEUTSCH, C
中科院分区：
文献类型：
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作者：
BENTAL, M;DEUTSCH, C
使用NMR光谱，我们研究了纯化的人T淋巴细胞在无血清培养基中。将纯化的细胞包埋在琼脂糖珠内，并通过促分裂剂佛波醇-12-肉豆蔻酸酯-13-乙酸酯和离子霉素诱导增殖。标准培养物和琼脂糖珠内的T淋巴细胞表现出相当的活力，DNA合成和白细胞介素-2分泌的程度和动力学相似。31 P-NMR显示刺激两天或更长时间的细胞中磷酸单酯减少，磷酸二酯含量增加。13 C-葡萄糖利用率和13 C-乳酸生产率显示85%的利用的葡萄糖被转化为乳酸盐。灌注介质的1H-NMR光谱表明，乳酸盐也从葡萄糖或糖原以外的底物产生。葡萄糖占静止细胞产生的乳酸的25%，并且占刺激细胞产生的乳酸的67%。糖酵解在刺激后的前2小时内增强6倍，到48或96小时增强15倍。到48小时，有氧乳酸盐产量增加了3倍，在刺激的前12小时仅略有增加。我们的研究结果表明，在细胞周期进程中，T淋巴细胞在G 0到G1过渡的前90分钟内产生的乳酸从主要是有氧的转变为主要是厌氧的。
Using NMR spectroscopy, we studied purified, human T lymphocytes in a serum‐free medium. Purified cells were entrapped inside agarose beads and induced to proliferate by the mitogens phorbol‐12‐myristate‐13‐acetate and ionomycin. T lymphocytes in standard culture and inside agarose beads exhibit comparable viability, and similar extent and kinetics of DNA synthesis and Interleukin‐2 secretion.31P‐NMR revealed decreased phosphomonoester and increased phosphodiester content in cells stimulated for two days or longer.13C‐glucose utilization and13C‐lactate production rates showed that 85% of the utilized glucose was converted to lactate.1H‐NMR spectra of the perfusing media indicated that lactate was also produced from substrates other than glucose or glycogen. Glucose accounted for 25% of the lactate produced by quiescent cells, and for 67% of lactate production by stimulated cells. Glycolysis was enhanced 6‐fold within the first 2 hours following stimulation, and 15‐fold by 48 or 96 h. Aerobic lactate production was increased 3‐fold by 48 h, with only a minor enhancement during the first 12 h of stimulation. Our results indicate a shift from mostly aerobic to mostly anaerobic lactate production in T lymphocytes within the first 90 min of theG0toG1transition during cell cycle progression.