Institute of Pathology, Southwest Hospital Affiliated to Third Military Medical University, Chongqing 400038, China.
Vaccine. 2011 May 31;29(24):4162-72. doi: 10.1016/j.vaccine.2011.03.056. Epub 2011 Apr 5.
The objectives of this study were to detect the immunogenicity of a lipopolysaccharide (LPS) binding protein (LBP) mimic epitope peptide vaccine and evaluate its effect on controlling excessive and uncontrolled inflammatory reactions in mice with acute lung injury. The vaccine was prepared by mixing self-made LBP mimic epitope multiple antigen peptide (MAP) and Freund adjuvants in a proper proportion. Healthy mice were inoculated with the vaccine and the dynamic changes of anti-MAP antibody were measured using ELISA. Anti-MAP antibody was prepared from the immune serum of the mice based on the standard antibody preparation program. Western blot assay was used to identify LBP specificity of anti-MAP antibody. Anti-MAP antibody bioactivity was analyzed using in vitro binding activity test. Following the vaccine inoculation, the mice were injected with LPS to induce acute lung injury. Anti-MAP antibody prepared was also used to immune the mice with LPS-induced acute lung injury. TNF-α and IL-1β contents in serum and lung tissue homogenate were measured using double antibody sandwiched ELISA at different time-points after LPS challenge. At 8h time-point, total white blood cell counts, polymorphonuclear leucocyte count and protein content in bronchoalveolar lavage fluid were measured and pulmonary morphological changes were evaluated. The antibody titer was gradually rising, reaching to its peak at 8th week and lasting to the tenth week. The antibody possessed strong immunogenicity, high specificity and favorable biologic activity. Whole range inoculation of LBP mimic epitope peptide vaccine or anti-MAP antibody intervention partly eliminates LPS-mediated acute lung injury. In conclusion, LBP mimic epitope peptide vaccine successfully induced highly specific antibody with high bioactivity in mice. LBP mimic epitope peptide vaccine and anti-MAP antibody inhibited excessive and uncontrolled inflammatory reactions from LPS-mediated acute lung injury in mice.
本研究的目的是检测脂多糖（LPS）结合蛋白（LBP）模拟表位肽疫苗的免疫原性，并评价其对控制脂多糖诱导的急性肺损伤小鼠过度和失控性炎症反应的作用。该疫苗通过将自制的 LBP 模拟表位多抗原肽（MAP）与弗氏佐剂按适当比例混合制备。将健康小鼠接种疫苗，采用 ELISA 法检测抗-MAP 抗体的动态变化。根据标准抗体制备程序，从免疫小鼠的血清中制备抗-MAP 抗体。采用 Western blot 法鉴定抗-MAP 抗体的 LBP 特异性。采用体外结合活性试验分析抗-MAP 抗体的生物活性。疫苗接种后，用 LPS 对小鼠进行注射以诱导急性肺损伤。用 LPS 诱导的急性肺损伤小鼠也用抗-MAP 抗体进行免疫。在 LPS 攻击后不同时间点，采用双抗体夹心 ELISA 法测定血清和肺组织匀浆中 TNF-α和 IL-1β含量。在 8h 时间点，测定支气管肺泡灌洗液中总白细胞计数、多形核白细胞计数和蛋白含量，并评价肺组织形态学变化。抗体滴度逐渐升高，第 8 周达到峰值，并持续到第 10 周。该抗体具有较强的免疫原性、高特异性和良好的生物活性。全范围接种 LBP 模拟表位肽疫苗或抗-MAP 抗体干预部分消除了 LPS 介导的急性肺损伤。综上所述，LBP 模拟表位肽疫苗成功诱导了具有高生物活性的高度特异性抗体。LBP 模拟表位肽疫苗和抗-MAP 抗体抑制了 LPS 介导的急性肺损伤小鼠过度和失控性炎症反应。
