Battafarano R J, Burd R S, Kurrelmeyer K M, Ratz C A, Dunn D L
Department of Surgery, University of Minnesota, Minneapolis.
Arch Surg. 1994 Feb;129(2):179-80. doi: 10.1001/archsurg.1994.01420260075010.
This study tried to determine whether administration of antilipopolysaccharide (LPS) murine monoclonal antibody (mAb) 2A3 to mice was associated with (1) protective capacity during experimental gram-negative bacterial sepsis, and (2) inhibition of tumor necrosis factor alpha (TNF-alpha) secretion in the systemic circulation and at the tissue level during experimental infection.
Mice received an initial intravenous injection of either saline or 100 micrograms of anti-LPS mAb 2A3, and 1 hour later underwent intraperitoneal inoculation of viable Escherichia coli 0111:B4. Mortality was assessed daily for 7 days. Separate groups of mice were treated similarly and plasma TNF-alpha concentrations were determined from blood samples obtained at 1, 3, 6, 10, and 16 hours after infection by enzyme-linked immunosorbent assay. Concurrently, splenocytes harvested from animals 3, 10, and 16 hours after infection were incubated in culture ex vivo and supernatant TNF-alpha levels were determined.
Pretreatment with anti-LPS mAb 2A3 prior to an intraperitoneal challenge of live E coli 0111:B4 was associated with the following: (1) significant protective capacity (100% vs 0% mortality, P < .001); (2) inhibition of plasma TNF-alpha levels 16 hours after infection (1257 +/- 323 pg/mL vs 292 +/- 254 pg/mL, P < .001); and (3) abrogation of TNF-alpha secretion derived from splenic macrophages isolated 16 hours after bacterial challenge (229 +/- 12 pg/mL vs 107 +/- 48 pg/mL, P < .05).
These results strongly support the contention that inhibition of LPS-induced TNF-alpha secretion at both the tissue and systemic levels is a key mechanism by which anti-LPS mAbs provide protection during gram-negative bacterial peritonitis. We believe that in vivo monitoring of macrophage cytokine secretion will be critical for elucidating the precise role of a variety of mediators in the pathogenesis of gram-negative bacterial sepsis.
本研究旨在确定给小鼠注射抗脂多糖（LPS）鼠单克隆抗体（mAb）2A3是否与以下情况相关：（1）在实验性革兰氏阴性菌败血症期间的保护能力；（2）在实验性感染期间，全身循环和组织水平上肿瘤坏死因子α（TNF-α）分泌的抑制情况。
小鼠最初静脉注射生理盐水或100微克抗LPS mAb 2A3，1小时后腹腔接种活的大肠杆菌0111:B4。连续7天每日评估死亡率。对另一组小鼠进行类似处理，并通过酶联免疫吸附测定法，测定感染后1、3、6、10和16小时采集的血样中的血浆TNF-α浓度。同时，将感染后3、10和16小时从动物体内采集的脾细胞进行体外培养，并测定培养上清液中的TNF-α水平。
在腹腔注射活的大肠杆菌0111:B4之前，用抗LPS mAb 2A3进行预处理与以下情况相关：（1）显著的保护能力（死亡率100%对0%，P <.001）；（2）感染后16小时血浆TNF-α水平受到抑制（1257±323 pg/mL对292±254 pg/mL，P <.001）；（3）细菌攻击后16小时分离的脾巨噬细胞分泌的TNF-α被消除（229±12 pg/mL对107±48 pg/mL，P <.05）。
这些结果有力地支持了以下观点，即在组织和全身水平上抑制LPS诱导的TNF-α分泌是抗LPS mAb在革兰氏阴性菌腹膜炎期间提供保护的关键机制。我们认为，体内监测巨噬细胞细胞因子分泌对于阐明多种介质在革兰氏阴性菌败血症发病机制中的精确作用至关重要。
