Kaneene J M, Anderson R K, Johnson D W, Muscoplat C C, Nicoletti P, Angus R D, Pietz D E, Klausner D J
J Clin Microbiol. 1978 Jun;7(6):550-7. doi: 10.1128/jcm.7.6.550-557.1978.
A study was conducted to develop an in vitro whole-blood lymphocyte stimulation assay for measurement of cell-mediated immune response in bovine brucellosis. A soluble antigen (BASA) prepared from killed cells of Brucella abortus 1119-3 was used. Cattle infected with B. abortus field strains, B. abortus 19 calfhood- and adult-vaccinated cattle, and nonexposed cattle were tested. Blood was diluted 10-fold in RPMI-1640 medium (without added serum) and cultured with BASA (at a concentration of 2.2 microgram per culture) at varying times of incubation. Results were assayed for [3H]thymidine incorporation into deoxyribonucleic acid. A 6-day period was found to be optimal for incubating blood cultures to achieve maximum specific lymphocyte stimulation. Serological tests and bacteriological isolation attempts were conducted simultaneously with lymphocyte stimulation tests, and there was a significant correlation between cell-mediated immune response and bacteriological findings. There was a significant correlation between cell-mediated immune response and the level of serum antibodies on a group basis, but there was little correlation between the two systems on individual infected animals. Among vaccinated animals there was little or no correlation between cell-mediated immune and humoral responses. The whole-blood assay was found to be simple, fast, sensitive, and reproducible.
开展了一项研究，以开发一种体外全血淋巴细胞刺激试验，用于测量牛布鲁氏菌病的细胞介导免疫反应。使用从流产布鲁氏菌1119-3的死菌细胞制备的可溶性抗原（BASA）。对感染了流产布鲁氏菌田间菌株的牛、接种过19号流产布鲁氏菌的犊牛和成牛以及未接触过该病的牛进行了检测。血液在RPMI-1640培养基（不添加血清）中稀释10倍，并在不同孵育时间与BASA（每培养物浓度为2.2微克）一起培养。检测结果为[3H]胸腺嘧啶核苷掺入脱氧核糖核酸的情况。发现6天的孵育期最适合全血培养，以实现最大程度的特异性淋巴细胞刺激。在进行淋巴细胞刺激试验的同时进行了血清学检测和细菌学分离尝试，细胞介导免疫反应与细菌学检测结果之间存在显著相关性。在群体基础上，细胞介导免疫反应与血清抗体水平之间存在显著相关性，但在个体感染动物中，这两个系统之间几乎没有相关性。在接种疫苗的动物中，细胞介导免疫反应与体液反应之间几乎没有相关性。发现全血试验简单、快速、灵敏且可重复。
