Stokol Tracy, Thomas Sophie Isabella, Hoffman Martha, Zhao Shay
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
Front Vet Sci. 2024 Aug 19;11:1405297. doi: 10.3389/fvets.2024.1405297. eCollection 2024.
CD80, a co-stimulatory molecule required for optimal T cell activation, is expressed on antigen-presenting cells, including monocytes and dendritic cells, in dogs and humans. We hypothesized that CD80 would be expressed on tumor cells in dogs from acute myeloid leukemia (AML) but not dogs with lymphoid neoplasms.
We first evaluated the cellular staining pattern of a hamster anti-murine CD80 antibody (clone 16-10A1, ThermoFisher Scientific Cat# 17-0801-82, RRID: AB_469417) in blood and bone marrow aspirates from healthy dogs. Using flow cytometric analysis and examination of modified Wright's-stained cytologic smears of unsorted and flow cytometric or immunomagnetic bead-sorted leukocytes, we show that the antibody binds to mature and immature neutrophils and monocytes, but not lymphocytes or eosinophils, in blood and bone marrow. We then added the antibody to routine flow cytometric panels for immunophenotyping hematopoietic neoplasms in dogs. We found that the antibody labeled tumor cells in 72% of 39 dogs with AML and 36% of 11 dogs with acute leukemia expressing lymphoid and myeloid markers ("mixed lineage") but none of the dogs with B ( = 37) or T ( = 3) lymphoid neoplasms. A higher proportion of tumor cells in dogs with AML were labeled with the anti-CD80 antibody vs antibodies against other myeloid-associated antigens, including CD4 (36%, = 0.003), CD11b (44%), CD11c (46%), CD14 (38%, = 0.006) and CD18 (59%, clone YFC118). In contrast, antibodies against CD11b and CD11c bound to tumor cells in 8-32% of the lymphoid neoplasms.
We show that CD80, as detected by antibody clone 16-10A1, is a sensitive and specific marker for AML and would be useful to include in flow cytometric immunophenotyping panels in dogs.
CD80是最佳T细胞活化所需的共刺激分子，在犬类和人类的抗原呈递细胞（包括单核细胞和树突状细胞）上表达。我们假设CD80会在患有急性髓性白血病（AML）的犬类肿瘤细胞上表达，但在患有淋巴肿瘤的犬类中不表达。
我们首先评估了仓鼠抗小鼠CD80抗体（克隆号16 - 10A1，赛默飞世尔科技产品编号17 - 0801 - 82，RRID: AB_469417）在健康犬类血液和骨髓穿刺物中的细胞染色模式。通过流式细胞术分析以及对未分选和经流式细胞术或免疫磁珠分选的白细胞的改良瑞氏染色细胞学涂片进行检查，我们发现该抗体与血液和骨髓中的成熟及未成熟中性粒细胞和单核细胞结合，但不与淋巴细胞或嗜酸性粒细胞结合。然后，我们将该抗体添加到用于犬类造血肿瘤免疫表型分析的常规流式细胞术检测板中。我们发现，在39只患有AML的犬类中，72%的肿瘤细胞被该抗体标记；在11只表达淋巴和髓系标志物（“混合谱系”）的急性白血病犬类中，36%的肿瘤细胞被标记；而在37只B淋巴细胞肿瘤犬类和3只T淋巴细胞肿瘤犬类中，无一例肿瘤细胞被标记。与针对其他髓系相关抗原（包括CD4（36%，P = 0.003）、CD11b（44%）、CD11c（46%）、CD14（38%，P = 0.006）和CD18（59%，克隆号YFC118））的抗体相比，患有AML犬类中的肿瘤细胞被抗CD80抗体标记的比例更高。相比之下，针对CD11b和CD11c的抗体在8% - 32%的淋巴肿瘤中与肿瘤细胞结合。
我们表明，通过抗体克隆号16 - 10A1检测到的CD80是AML的一种敏感且特异的标志物，将其纳入犬类流式细胞术免疫表型分析检测板会很有用。
