Concha C, Holmberg O
National Veterinary Institute, Swedish University of Agricultural Sciences, Uppsala.
J Dairy Res. 1990 Feb;57(1):7-16. doi: 10.1017/s0022029900026558.
Cells were obtained by centrifuging the mammary secretion of healthy udders of 19 cows during the dry-period and during mid-lactation. The suspended cells were incubated in plastic wells. Those adhered cells classified as mammary macrophages were incubated with pokeweed mitogen (PWM). Autologous peripheral blood lymphocytes were added to wells containing untreated macrophage cultures or cultures pretreated with PWM. In seven cows autologous dry-period mammary lymphocytes were added instead of blood lymphocytes. The macrophages + lymphocyte cultures were subjected to the lymphocyte stimulation test (LST). For comparison, peripheral blood lymphocytes and dry-period secretion lymphocytes were also subjected to the LST in the presence of PWM. In all cases, mitogenic responses were higher in pretreated macrophage cultures than in background control cultures. The stimulation indices (SI) showed that PWM-pretreated dry-period mammary macrophages enhanced the proliferation of autologous peripheral blood lymphocytes to a greater extent than did blood lymphocytes plus PWM (49 +/- 10 v. 30 +/- 6; P less than or equal to 0.05). Mammary macrophages taken from the same cows but during midlactation also clearly induced proliferation of autologous peripheral blood lymphocytes but to a lesser extent than dry-period macrophages (16 +/- 2 v. 49 +/- 10; 16 +/- 2 v. 30 +/- 6; P less than or equal to 0.01 and P less than or equal to 0.05). The PWM pretreatment of mammary macrophages increased the proliferation of autologous dry-period mammary lymphocytes by at least a factor of three (28 +/- 8 v. 8 +/- 2 P less than or equal to 0.05). The present results indicate that bovine mammary macrophages pretreated with PWM enhance proliferation as well as modulation of mammary and peripheral blood lymphocytes. The modulation of lymphocyte stimulation as demonstrated here in vitro, has great significance regarding aspects of local immunostimulation related to modern treatment of mastitis.
通过对19头奶牛干奶期和泌乳中期健康乳房的乳汁进行离心获取细胞。将悬浮细胞接种于塑料孔板中。将那些被归类为乳腺巨噬细胞的贴壁细胞与商陆有丝分裂原（PWM）一起孵育。将自体外周血淋巴细胞加入含有未处理巨噬细胞培养物或经PWM预处理的培养物的孔板中。在7头奶牛中，加入自体干奶期乳腺淋巴细胞替代血液淋巴细胞。巨噬细胞+淋巴细胞培养物进行淋巴细胞刺激试验（LST）。作为对照，外周血淋巴细胞和干奶期分泌淋巴细胞也在PWM存在的情况下进行LST。在所有情况下，预处理巨噬细胞培养物中的促有丝分裂反应均高于背景对照培养物。刺激指数（SI）显示，PWM预处理的干奶期乳腺巨噬细胞比血液淋巴细胞加PWM更能增强自体外周血淋巴细胞的增殖（49±10对30±6；P≤0.05）。取自同一奶牛但处于泌乳中期的乳腺巨噬细胞也明显诱导自体外周血淋巴细胞增殖，但程度低于干奶期巨噬细胞（16±2对49±10；16±2对30±6；P≤0.01和P≤0.05）。乳腺巨噬细胞的PWM预处理使自体干奶期乳腺淋巴细胞的增殖至少增加了三倍（28±8对8±2；P≤0.05）。目前的结果表明，经PWM预处理的牛乳腺巨噬细胞可增强乳腺和外周血淋巴细胞的增殖及调节作用。本文体外实验所证明的淋巴细胞刺激调节作用，对于与乳腺炎现代治疗相关的局部免疫刺激方面具有重要意义。
