Bakker W W, van Luijk W H, Hené R J, Desmit E M, van der Hem G K, Vos J T
Am J Nephrol. 1986;6(2):107-11. doi: 10.1159/000167064.
Peripheral mononuclear blood cells isolated from nephrotic subjects with minimal-change nephrotic syndrome (selective proteinuria greater than 3.5 g/24 h) or various other forms of glomerulonephritis (non-selective proteinuria greater than 3.5 g/24 h) were stimulated with concanavalin A and cultured for 20 h in the presence of kidney tissue under standard conditions. Identical cultures were developed with phosphate-buffered saline from normal control donors. Triplicate cultures of each subject (3 X 10(6) cells/ml) were incubated with or without 5, 10, or 20 micrograms/ml concanavalin A per milliliter serum-free tissue culture medium upon cryostat sections from normal rat kidney. The cells were subsequently removed, and the tissue sections were washed and stained for sialoprotein using the colloidal iron method and evaluated for stainability of glomerular polyanion using light microscopy. The results show that peripheral mononuclear blood cells from subjects with minimal-change nephrotic syndrome had affected glomerular polyanion in vitro during incubation with kidney tissue in a significantly (p less than or equal to 0.005) higher number of cases (15/17) as compared with the number of glomerulonephritis patients who scored positive in 4 out of 14 cases, whereas this was the case in 3 out of 18 cases of the normal donors. It is concluded that stimulated cellular immune reactivity of peripheral mononuclear blood cells from subjects with minimal-change nephrotic syndrome in vitro is associated with the potential impairment in vitro of an important part of the glomerular filtration barrier. Since this cellular activity occurred to a significant lesser extent in other nephrotic subjects, this response is not related to the nephrotic state per se.
从患有微小病变肾病综合征(选择性蛋白尿大于3.5g/24小时)或各种其他形式肾小球肾炎(非选择性蛋白尿大于3.5g/24小时)的肾病患者中分离出外周血单个核细胞,用伴刀豆球蛋白A刺激,并在标准条件下于肾组织存在的情况下培养20小时。用来自正常对照供体的磷酸盐缓冲盐水建立相同的培养物。将每个受试者的一式三份培养物(3×10⁶个细胞/ml)在不含血清的组织培养基中,每毫升加入或不加入5、10或20微克/ml伴刀豆球蛋白A,与正常大鼠肾脏的冰冻切片一起孵育。随后去除细胞,将组织切片洗涤并用胶体铁法对涎蛋白进行染色,并使用光学显微镜评估肾小球多阴离子的可染性。结果显示,与14例中有4例呈阳性的肾小球肾炎患者相比,微小病变肾病综合征患者的外周血单个核细胞在与肾组织孵育期间,在体外影响肾小球多阴离子的病例数显著更多(15/17,p≤0.005),而正常供体的18例中有3例出现这种情况。得出结论,微小病变肾病综合征患者外周血单个核细胞的刺激细胞免疫反应性在体外与肾小球滤过屏障重要部分的体外潜在损伤相关。由于这种细胞活性在其他肾病患者中发生的程度要小得多,这种反应与肾病状态本身无关。