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用于在冷冻及福尔马林固定石蜡包埋组织中对猫B淋巴细胞进行免疫组织化学染色的单克隆抗体的鉴定

Identification of monoclonal antibodies for immunohistochemical staining of feline B lymphocytes in frozen and formalin-fixed paraffin-embedded tissues.

作者信息

Monteith C E, Chelack B J, Davis W C, Haines D M

机构信息

Department of Pathology, Western College of Veterinary Medicine, Saskatoon, Saskatchewan.

出版信息

Can J Vet Res. 1996 Jul;60(3):193-8.

Abstract

Commercially-available monoclonal antibodies to B lymphocytes were evaluated for immunohistochemical staining of feline B lymphocytes in frozen and formalin-fixed, paraffin-embedded tissues using an avidin biotin complex immunoperoxidase immunohistochemical technique. Three monoclonal antibodies: F46A and F72A raised to "carnivore" B lymphocytes and RA3.6B2 raised to murine B lymphocytes, stained B lymphocyte-dependent areas of frozen feline lymphoid tissue. In addition, antibody RA3.6B2 stained B lymphocyte dependent areas in formalin-fixed, paraffin-embedded feline tissues. There was no staining of T lymphocyte-dependent areas in either frozen or formalin-fixed tissues. Dual parameter flow cytometry, using an anti-pan-T lymphocyte antibody, revealed that greater than 99% of the cells stained by RA3.6B2 are a population distinct from T lymphocytes. F46A was shown to stain a sub-population of those cells stained with RA3.6B2. These antibodies may be useful in the identification of feline B lymphocytes using immunohistochemistry and flow cytometry and thereby provide additional tools to study B lymphocyte ontogeny and the significance of lymphocyte phenotype in lymphoid neoplasia in cats.

摘要

利用抗生物素蛋白-生物素复合物免疫过氧化物酶免疫组织化学技术,对市售的抗B淋巴细胞单克隆抗体进行评估,以检测其在冷冻及福尔马林固定、石蜡包埋组织中对猫B淋巴细胞的免疫组织化学染色效果。三种单克隆抗体:针对“食肉动物”B淋巴细胞产生的F46A和F72A,以及针对鼠B淋巴细胞产生的RA3.6B2,均能对冷冻猫淋巴组织中B淋巴细胞依赖区进行染色。此外,抗体RA3.6B2还能对福尔马林固定、石蜡包埋的猫组织中的B淋巴细胞依赖区进行染色。在冷冻或福尔马林固定的组织中,T淋巴细胞依赖区均未出现染色。使用抗全T淋巴细胞抗体进行双参数流式细胞术检测发现,RA3.6B2染色的细胞中超过99%是与T淋巴细胞不同的细胞群体。结果显示,F46A能对那些被RA3.6B2染色的细胞中的一个亚群进行染色。这些抗体可能有助于利用免疫组织化学和流式细胞术鉴定猫B淋巴细胞,从而为研究猫B淋巴细胞个体发生以及淋巴细胞表型在猫淋巴肿瘤中的意义提供更多工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0423/1263832/7f30c98ea1a9/cjvetres00023-0038-a.jpg

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