Tingstedt Jens-Erik, Tornehave Ditte, Lind Peter, Nielsen Jens
Department of Virology, Danish Veterinary Institute, Lindholm, DK-4771, Kalvehave, Denmark.
Vet Immunol Immunopathol. 2003 Aug 15;94(3-4):123-32. doi: 10.1016/s0165-2427(03)00096-5.
Identification of the different cell types of the immune system is important for in situ studies on the pathogenesis of infectious diseases in various animals, including the pig. Unfortunately, many monoclonal anti-leukocyte antibodies are only useful for staining frozen tissue sections with inherent poor tissue morphology, and are not readily adapted to formaldehyde fixed and paraffin embedded tissue with well preserved morphology. Seven well characterised monoclonal antibodies against porcine leukocyte antigens were tested on neutral buffered paraformaldehyde fixed and paraffin embedded porcine tissue sections using the highly sensitive tyramide signal amplification system. Combining this method with different antigen retrieval techniques enabled us to detect CD2, CD3, CD4, CD8 and SWC3 antigen expressing cells in porcine lymphoid tissue. Thus, we describe herein methods for the detection of several major cell types of the porcine immune system in fixed tissue with optimal preservation of histological details.
识别免疫系统的不同细胞类型对于包括猪在内的各种动物传染病发病机制的原位研究很重要。不幸的是,许多单克隆抗白细胞抗体仅适用于对组织形态固有较差的冷冻组织切片进行染色,并且不易适用于形态保存良好的甲醛固定石蜡包埋组织。使用高灵敏度酪胺信号放大系统,对七种针对猪白细胞抗原的特征明确的单克隆抗体在中性缓冲多聚甲醛固定石蜡包埋的猪组织切片上进行了测试。将该方法与不同的抗原修复技术相结合,使我们能够在猪淋巴组织中检测到表达CD2、CD3、CD4、CD8和SWC3抗原的细胞。因此,我们在此描述了在固定组织中检测猪免疫系统几种主要细胞类型的方法,同时能最佳地保存组织学细节。