von Asmuth E J, Leeuwenberg J F, Ceska M, Buurman W A
Department of Surgery, University of Limburg, Maastricht, The Netherlands.
Eur Cytokine Netw. 1991 Aug-Sep;2(4):291-7.
In this in vitro study, the influence of serum-concentration, heat inactivation of the serum and the origin of the serum on the responsiveness of cultured human umbilical vein endothelial cells (HUVEC) to immunological challenges was investigated. Addition of human serum during stimulation with 1 microgram/ml bacterial lipopolysaccharide (LPS) increased endothelial cell ELAM-1 expression and interleukin (IL)-6 release five to ten-fold. Full endothelial cell responsiveness to LPS required 10 to 50% human serum and was largely abrogated after heating the serum for 30 minutes at 56 degrees C. Addition of newborn or fetal bovine serum instead of human serum, induced even higher IL-6 release and ELAM-1 expression in response to LPS, whilst heat-inactivation of these serum-batches only moderately decreased endothelial cell responses. Endothelial cell IL-6 release and ELAM-1 expression after stimulation with IL-1 beta and tumor necrosis factor-alpha (TNF-alpha) were less influenced by heat inactivation of the serum and by omission of serum, whilst responses to PMA remained completely unaffected by such modifications in assay media. Finally, we demonstrated that endothelial cell IL-8 release also and ICAM-1 expression in response to LPS and cytokines were increased by addition of human serum, indicating that the use of serum-free assay media, or the use of media enriched with heat-inactivated (HI) human serum interferes with physiological endothelial cell responsiveness.(ABSTRACT TRUNCATED AT 250 WORDS)
在这项体外研究中，研究了血清浓度、血清热灭活以及血清来源对培养的人脐静脉内皮细胞（HUVEC）免疫应答反应性的影响。在用1微克/毫升细菌脂多糖（LPS）刺激期间添加人血清，可使内皮细胞ELAM-1表达和白细胞介素（IL）-6释放增加5至10倍。内皮细胞对LPS的完全反应性需要10%至50%的人血清，并且在56℃将血清加热30分钟后反应性基本消除。添加新生牛或胎牛血清而非人血清，可诱导对LPS产生更高的IL-6释放和ELAM-1表达，而这些血清批次的热灭活仅适度降低内皮细胞反应。在用IL-1β和肿瘤坏死因子-α（TNF-α）刺激后，内皮细胞IL-6释放和ELAM-1表达受血清热灭活和血清缺失的影响较小，而对佛波酯（PMA）的反应完全不受检测培养基此类改变的影响。最后，我们证明添加人血清也可增加内皮细胞对LPS和细胞因子的IL-8释放以及ICAM-1表达，表明使用无血清检测培养基或使用富含热灭活（HI）人血清的培养基会干扰内皮细胞的生理反应性。（摘要截选至250字）
