Caperna T J, Garvey J S
Mech Ageing Dev. 1982 Nov;20(3):205-21. doi: 10.1016/0047-6374(82)90088-4.
A method has been developed for the purification of Kupffer and endothelial cells from rat liver by collagenase enzyme perfusion followed by centrifugal elutriation. After intravenous injection of a soluble antigen, [3H]azoaniline bovine serum albumin ( [3H]BSA), its distribution was studied in isolated cell populations from liver and spleen tissue of two aged groups of male F-344 rats. In young adult rats (6-8 months) both sinusoidal cell types contained the same amount of [3H]BSA; however, in older rats (22-24 months) the amount of antigen in the endothelial cells was significantly decreased. In comparison to the liver, the spleen retained only a small fraction of the injected dose. In order to assess the catabolic properties of both Kupffer and endothelial cells, supernatants obtained from in vitro cell culture were evaluated for both biological and physiochemical properties. Antigen was almost completely degraded by both cell types as determined by gel filtration and did not directly stimulate BSA-primed lymphocytes in vitro; however, these supernatants were shown to enhance the lymphoproliferative response of primed lymphocytes to additional antigen exposure. Kupffer cell receptors, Fc and C3, assayed by direct rosetting, did not vary with age; endothelial cells also possessed Fc receptors that were found to be unchanged with age. These studies are an initial attempt to better define our previous finding of defective antigen handling with aging by use of isolated pure cell populations.
已开发出一种方法,通过胶原酶灌注随后离心淘析从大鼠肝脏中纯化枯否细胞和内皮细胞。静脉注射可溶性抗原[³H]偶氮苯胺牛血清白蛋白([³H]BSA)后,研究了其在两组老年雄性F-344大鼠肝脏和脾脏组织分离细胞群体中的分布。在年轻成年大鼠(6 - 8个月)中,两种窦状隙细胞类型所含的[³H]BSA量相同;然而,在老年大鼠(22 - 24个月)中,内皮细胞中的抗原量显著减少。与肝脏相比,脾脏仅保留了注射剂量的一小部分。为了评估枯否细胞和内皮细胞的分解代谢特性,对体外细胞培养获得的上清液进行了生物学和物理化学性质评估。通过凝胶过滤测定,两种细胞类型几乎完全降解了抗原,并且在体外不会直接刺激经BSA致敏的淋巴细胞;然而,这些上清液显示可增强致敏淋巴细胞对额外抗原暴露的淋巴细胞增殖反应。通过直接玫瑰花结试验测定的枯否细胞受体Fc和C3不随年龄变化;内皮细胞也具有Fc受体,且发现其不随年龄变化。这些研究是首次尝试通过使用分离的纯细胞群体来更好地界定我们之前关于衰老导致抗原处理缺陷的发现。