Highland Margaret A, Schneider David A, White Stephen N, Madsen-Bouterse Sally A, Knowles Donald P, Davis William C
Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164-7040, USA; Animal Disease Research Unit, United States Department of Agriculture, Pullman, WA 99164-6630, USA.
Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, WA 99164-7040, USA.
Comp Immunol Microbiol Infect Dis. 2016 Jun;46:40-6. doi: 10.1016/j.cimid.2016.04.006. Epub 2016 Apr 14.
Although both domestic sheep (DS) and bighorn sheep (BHS) are affected by similar respiratory bacterial pathogens, experimental and field data indicate BHS are more susceptible to pneumonia. Cross-reactive monoclonal antibodies (mAbs) for use in flow cytometry (FC) are valuable reagents for interspecies comparative immune system analyses. This study describes cross-reactive mAbs that recognize leukocyte differentiation molecules (LDMs) and major histocompatibility complex antigens on DS and BHS leukocytes. Characterization of multichannel eosinophil autofluorescence in this study permitted cell-type specific gating of granulocytes for evaluating LDMs, specifically on neutrophils, by single-label FC. Evaluation of relative abundances of LDMs by flow cytometry revealed greater CD11a, CD11b, CD18 (β2 integrins) and CD 172a (SIRPα) on DS neutrophils and greater CD14 (lipopolysaccharide receptor) on BHS neutrophils. Greater CD25 (IL-2) was identified on BHS lymphocytes following Concavalin A stimulation. While DS and BHS have similar total peripheral blood leukocyte counts, BHS have proportionately more neutrophils.
虽然家养绵羊(DS)和大角羊(BHS)都会受到相似的呼吸道细菌病原体的影响,但实验和现场数据表明,大角羊更容易患肺炎。用于流式细胞术(FC)的交叉反应性单克隆抗体(mAb)是种间比较免疫系统分析的宝贵试剂。本研究描述了识别家养绵羊和大角羊白细胞上白细胞分化分子(LDM)和主要组织相容性复合体抗原的交叉反应性单克隆抗体。本研究中对多通道嗜酸性粒细胞自发荧光的表征允许通过单标记流式细胞术对粒细胞进行细胞类型特异性门控,以评估LDM,特别是在中性粒细胞上。通过流式细胞术评估LDM的相对丰度发现,家养绵羊中性粒细胞上的CD11a、CD11b、CD18(β2整合素)和CD172a(信号调节蛋白α)更多,而大角羊中性粒细胞上的CD14(脂多糖受体)更多。在刀豆蛋白A刺激后,大角羊淋巴细胞上的CD25(白细胞介素-2)更多。虽然家养绵羊和大角羊的外周血白细胞总数相似,但大角羊的中性粒细胞比例更高。