Claesson M H, Tscherning T, Nissen M H, Lind K
Department of Medical Anatomy A. University of Copenhagen, Denmark.
Scand J Immunol. 1990 Dec;32(6):623-30. doi: 10.1111/j.1365-3083.1990.tb03204.x.
Non-fermenting mycoplasma species deplete culture media for arginine through arginase activity linked to their arginine deiminase pathway, resulting in proliferation arrest and cell death in mycoplasma-contaminated cell cultures. The presence of only 2-3 Mycoplasma (M.) arginini-contaminated T cells in a one-way allogeneic mixed-lymphocyte culture (MLC) significantly inhibits development of cytotoxic T-cell activity. Likewise, strong degrees of inhibition are observed after addition of nanogram doses of M. arginini extracts (MAE) to MLC or cell proliferation cultures. M. arginini-induced cell inhibition can be reversed by addition of excess arginine to the culture medium. Antisera raised against non-fermenting, but not against fermenting, mycoplasma species block the inhibitory effect of MAE. SDS-PAGE separation of MAE disclosed a broad band at 60 kDa which contained arginase activity when assayed in MLC and cell proliferation culture. SDS-PAGE followed by western blotting and reaction with antisera raised against non-fermenting mycoplasma species demonstrated a band at 43 kDa common for these micro-organisms.
非发酵支原体通过与精氨酸脱亚胺酶途径相关的精氨酸酶活性消耗培养基中的精氨酸，导致支原体污染的细胞培养物增殖停滞和细胞死亡。在单向同种异体混合淋巴细胞培养（MLC）中，仅存在2 - 3个被精氨酸支原体（M. arginini）污染的T细胞就会显著抑制细胞毒性T细胞活性的发展。同样，在向MLC或细胞增殖培养物中添加纳克剂量的精氨酸支原体提取物（MAE）后，也观察到了强烈程度的抑制作用。通过向培养基中添加过量精氨酸，可以逆转精氨酸支原体诱导的细胞抑制作用。针对非发酵支原体而非发酵支原体产生的抗血清可阻断MAE的抑制作用。MAE的SDS - PAGE分离在60 kDa处显示出一条宽带，在MLC和细胞增殖培养物中检测时，该宽带具有精氨酸酶活性。SDS - PAGE后进行蛋白质印迹分析，并与针对非发酵支原体产生的抗血清反应，结果显示这些微生物在43 kDa处有一条共同的条带。
