Davis W C, Marusic S, Lewin H A, Splitter G A, Perryman L E, McGuire T C, Gorham J R
Vet Immunol Immunopathol. 1987 Jul;15(4):337-76. doi: 10.1016/0165-2427(87)90005-5.
We examined the potential of developing a set of species specific and cross reactive monoclonal antibodies (MoAbs) for use in the study of the phylogenetic and functional relation of class I and class II antigens of the major histocompatibility complex (MHC) and leukocyte differentiation antigens in cattle and other species. Comparing immunization strategies demonstrated the number of hybrids producing cross reactive antibodies can be increased by hyperimmunization of mice with lymphoid cells from multiple species. Comparing various methods of assay (antibody-complement mediated cytotoxicity [CT], enzyme linked immunosorbent assay [ELISA] and flow microfluorimetry [FMF]), revealed FMF is the most useful technique for the primary assay of hybridomas producing MoAbs of potential interest. By using dual parameter and dual fluorescence analysis, we could determine whether a given MoAb reacted with mononuclear cells (lymphocytes and monocytes) and/or granulocytes, and also whether any two MoAbs of different isotype and specificity recognized antigens present on identical or separate populations of leukocytes. Comparing the patterns of MoAb reactivity with leukocytes obtained from cows, goats, sheep, pigs, horses and humans, as well as comparing the patterns of reactivity with a panel of lymphoid cell lines derived from cattle (with enzootic bovine leukemia) and humans (with various forms of leukemia), revealed sets of MoAbs reactive with unique antigenic determinants present on BoLA class I (15 MoAbs) and class II (9 MoAbs) antigens, and also MoAbs reactive with determinants present on leukocyte differentiation antigens (36 MoAbs). Dual fluorescence analysis demonstrated the antigens detected by some MoAbs are predominantly expressed on one lineage of leukocytes while others are expressed on two or more lineages of leukocytes. Dual and single fluorescence analysis also demonstrated the PNA receptor(s) is: expressed on T cells, granulocytes and class II antigen monocytes and absent or expressed in low amount on sIgM+ B cells and a newly defined Non T/Non B population of cells. The strategies described for identifying and analyzing the specificity of MoAbs demonstrate the feasibility of developing a set of cross reactive MoAbs for identifying homologous molecules in multiple species and delineating their functional and phylogenetic relation.
我们研究了开发一组物种特异性和交叉反应性单克隆抗体（MoAb）的潜力，用于研究牛和其他物种主要组织相容性复合体（MHC）的I类和II类抗原以及白细胞分化抗原的系统发育和功能关系。比较免疫策略表明，通过用来自多个物种的淋巴细胞对小鼠进行超免疫，可以增加产生交叉反应性抗体的杂交瘤数量。比较各种检测方法（抗体-补体介导的细胞毒性[CT]、酶联免疫吸附测定[ELISA]和流式微荧光测定[FMF]），发现FMF是用于初步检测产生潜在感兴趣的MoAb的杂交瘤的最有用技术。通过使用双参数和双荧光分析，我们可以确定给定的MoAb是否与单核细胞（淋巴细胞和单核细胞）和/或粒细胞反应，以及不同同种型和特异性的任意两种MoAb是否识别存在于相同或不同白细胞群体上的抗原。比较从牛、山羊、绵羊、猪、马和人类获得的白细胞的MoAb反应模式，以及比较与一组来自牛（患有地方流行性牛白血病）和人类（患有各种形式白血病）的淋巴细胞系的反应模式，发现了与BoLA I类（15种MoAb）和II类（9种MoAb）抗原上存在的独特抗原决定簇反应的MoAb组，以及与白细胞分化抗原上存在的决定簇反应的MoAb（36种MoAb）。双荧光分析表明，一些MoAb检测到的抗原主要在一种白细胞谱系上表达，而其他抗原在两种或更多种白细胞谱系上表达。双荧光和单荧光分析还表明，PNA受体在T细胞、粒细胞和II类抗原单核细胞上表达，在sIgM+B细胞和新定义的非T/非B细胞群体上不存在或低表达。所描述的用于鉴定和分析MoAb特异性的策略证明了开发一组交叉反应性MoAb以鉴定多个物种中的同源分子并描绘其功能和系统发育关系的可行性。
