Kaneene J M, Johnson D W, Anderson R K, Angus R D, Pietz D E, Muscoplat C C
Am J Vet Res. 1978 Apr;39(4):585-9.
Cell-mediated immune responses in cattle naturally infected with strains of Brucella abortus and in cattle vaccinated with B abortus strain 19 during calfhood were studied by an in vitro lymphocyte-stimulation procedure. Lymphocytes were prepared from peripheral bovine blood by the Ficoll-diatrizoate technique, suspended in RPMI-1640 medium (1.5 X 10(6) lymphocytes/ml), cultured with B abortus-soluble antigen or phytohemagglutinin, and incubated for 6 days. Sixteen hours prior to termination of incubation, cultures were labeled with 1 muCi of [3H]thymidine (3HdT) and, after harvesting, assayed for 3HdT incorporation into DNA by liquid scintillation spectrometry. Lymphocytes from cattle with bacteriologically confirmed isolation of B abortus underwent a significantly higher lymphocyte stimulation with B abortus-soluble antigen than did cattle vaccinated with B abortus strain 19 during calfhood (P less than 0.005). Standard seroagglutination tests were conducted simultaneously with lymphocyte-stimulation tests, but there was no apparent correlation between levels of humoral antibodies and the cell-mediated immune responses as measured by in vitro specific lymphocyte stimulation.
采用体外淋巴细胞刺激法，对自然感染流产布鲁氏菌菌株的牛以及犊牛期接种流产布鲁氏菌19号菌株的牛的细胞介导免疫反应进行了研究。通过聚蔗糖-泛影葡胺技术从牛外周血中制备淋巴细胞，将其悬浮于RPMI-1640培养基中（1.5×10⁶个淋巴细胞/毫升），与流产布鲁氏菌可溶性抗原或植物血凝素一起培养，并孵育6天。在孵育结束前16小时，用1微居里的[³H]胸腺嘧啶核苷（³HdT）对培养物进行标记，收获后，通过液体闪烁光谱法测定³HdT掺入DNA的情况。经细菌学证实分离出流产布鲁氏菌的牛的淋巴细胞，与犊牛期接种流产布鲁氏菌19号菌株的牛相比，对流产布鲁氏菌可溶性抗原的淋巴细胞刺激反应明显更高（P＜0.005）。在进行淋巴细胞刺激试验的同时进行了标准血清凝集试验，但通过体外特异性淋巴细胞刺激测定的体液抗体水平与细胞介导免疫反应之间没有明显相关性。
