THE ESSENTIAL ROLE OF L-GLUTAMINE IN LYMPHOCYTE DIFFERENTIATION INVITRO
THE ESSENTIAL ROLE OF L-GLUTAMINE IN LYMPHOCYTE DIFFERENTIATION INVITRO
复制标题
DOI：
10.1002/jcp.1041240216
复制
发表时间：
1985-01-01
影响因子：
5.6
通讯作者：
COHEN, HJ
中科院分区：
文献类型：
--
作者：
CRAWFORD, J;COHEN, HJ
人类B淋巴细胞分化为浆细胞的生物化学机制尚不完全清楚。在商陆有丝分裂原刺激的人外周血单核细胞培养中，淋巴细胞转化和浆细胞形成似乎都需要L-谷氨酰胺。在含有10%热灭活和透析胎牛血清的GLN缺乏的RPMI1640中培养的细胞在72小时不能掺入~3H-胸腺嘧啶核苷或发生形态淋巴母细胞转化，仅用0.08 mM的L-谷氨酰胺或含APPRX的未透析热灭活胎牛血清可最大限度地恢复~3H-胸腺嘧啶核苷掺入。1 mm L-Gln.在随后的培养中，使用含有10%未透析热灭活胎牛血清的缺乏谷氨酰胺的RPMI1640，无论是否补充L-谷氨酰胺，淋巴母细胞转化都是相同的。只有添加2 mM L-谷氨酰胺的培养细胞才能进行浆细胞分化，用荧光素标记的抗Ig进行胞浆染色。用~3H-亮氨酸掺入免疫球蛋白来分析细胞免疫球蛋白合成和分泌的动力学时，加入2 mM L-谷氨酰胺的细胞合成和分泌分别增加2-5倍和3-10倍。免疫组织化学显示，只有添加谷氨酰胺的细胞粗面内质网发育与免疫球蛋白生成活跃一致。补充L-谷氨酰胺对细胞恢复、存活率、%B细胞、%T细胞、%单核细胞、%辅助性T细胞和抑制性T细胞均无明显影响。L-谷氨酰胺是淋巴细胞转化和浆细胞分化所必需的。未来对培养细胞的选择性营养需求的研究应该会对免疫细胞分化和功能的生化控制产生进一步的见解。
The biochemistry of human B lymphocyte differentiation to plasma cells is incompletely understood. L-Gln appears to be required for both lymphoblastic transformation and plasma cell formation in pokeweed-mitogen-stimulated human peripheral blood mononuclear cell cultures. Cells cultured with pokeweed mitogen in Gln-deficient RPMI-1640 with 10% heat-inactivated and dialyzed fetal bovine serum were unable to incorporate 3H-thymidine or undergo morphologic lymphoblastic transformation assessed at 72 h. 3H-thymidineincorporation could be maximally restored with as little as 0.08 mM L-Gln or by using nondialyzed heat-inactivated fetal bovine serum, containing .apprx. 1 mM L-Gln. In subsequent cultures, using Gln-deficient RPMI-1640 with 10% nondialyzed heat-inactivated fetal bovine serum, lymphoblastic transformation was equivalent with or without additional L-Gln supplementation. Only cultures with 2 mM L-Gln supplementation underwent plasma cell differentiation as assessed by cytoplasmic staining with fluorescein-conjugated anti-Ig. When the kinetics of cellular Ig synthesis and secretion were analyzed by 3H-Leu incorporation into Ig, synthesis was 2-5 fold greater, and secretion 3-10-fold greater in cell cultures with 2 mM L-Gln supplementation. By EM, only the Gln-supplemented cells showed development of rough endoplasmic reticulum consistent with active Ig production. L-Gln supplementation had no apparent effect on cell recovery, viability, % B cells, % T cells, % monocytes, or % helper and suppressor T cells. L-Gln is essential for both lymphoblastic transformation and plasma cell differentiation. Future investigation of the selective nutritional requirements of cultured cells should yield further insights into the biochemical control of immune cell differentiation and function.