Lewin H A, Calvert C C, Bernoco D
Am J Vet Res. 1985 Apr;46(4):785-8.
Ficoll-thrombin purified suspensions of bovine, equine, ovine, and porcine peripheral blood lymphocytes were fractionated on nylon-wool columns. The percentages of surface immunoglobulin (SIg+)-bearing lymphocytes in the adherent (B-cell enriched) and nonadherent (T-cell enriched) fractions were determined for individual animals using fluorescein isothiocyanate conjugated species-specific anti-Ig sera. Subsequently, the human leukocyte antigen DR-specific monoclonal antibody, H4, was tested for its ability to recognize a cross-reactive antigen on the fractionated lymphocytes, using the microcytotoxicity technique. The H4 plus complement killed a percentage of lymphocytes equivalent to the percentage of SIg+ lymphocytes in the adherent and nonadherent fractions. In a parallel experiment, a 2 fluorochrome technique was used to visualize bovine lymphocytes that were SIg+ and H4+. Lymphocytes that were SIg+ also stained with ethidium bromide (orange fluorescence) after complement-mediated cytotoxicity. Seemingly, H4 recognizes an evolutionarily conserved major histocompatibility complex encoded class-II-like determinant on the B lymphocytes of cattle, horses, sheep, and swine.
用Ficoll-凝血酶纯化牛、马、羊和猪外周血淋巴细胞悬液，然后在尼龙毛柱上进行分离。使用异硫氰酸荧光素偶联的种属特异性抗Ig血清，测定个体动物贴壁（富含B细胞）和非贴壁（富含T细胞）组分中携带表面免疫球蛋白（SIg+）的淋巴细胞百分比。随后，使用微量细胞毒性技术，检测人白细胞抗原DR特异性单克隆抗体H4识别分离淋巴细胞上交叉反应抗原的能力。H4加补体杀死的淋巴细胞百分比与贴壁和非贴壁组分中SIg+淋巴细胞的百分比相当。在一项平行实验中，使用双荧光染料技术观察SIg+和H4+的牛淋巴细胞。补体介导的细胞毒性作用后，SIg+淋巴细胞也用溴化乙锭染色（橙色荧光）。似乎，H4识别牛、马、羊和猪B淋巴细胞上由进化上保守的主要组织相容性复合体编码的类II样决定簇。
