Immunohistochemical identification of immune cell subsets in formalin- and zinc-fixed, paraffin-embedded tissues from chicken and duck using commercial antibodies
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引用次数: 0
Abstract
Immunohistochemical identification of immune cells in poultry has primarily been performed using frozen tissues, with limited identification in paraffin-embedded tissues. In this study, the following 18 commercially available primary antibodies associated with immune cell phenotypes were tested: anti-CD3, CD4 (clone CT-4 and 2‐35), TCRγδ, TCRαVβ1, TCRαVβ2, CD8, BAFF-R, PAX5, Bu-1a/b, Iba-1, MRC1L-B, CSF-1R, TIM4, MHC class II (clone 2D5 and 21‐1A6), MUM1, and CD45 antibodies in formalin-fixed, paraffin-embedded (FFPE) and zinc-fixed, paraffin-embedded (ZFPE) chicken and duck lymphoid tissues. In chickens, 11 antibodies in FFPE tissue and 16 in ZFPE tissue reacted with the expected antigens under some of the antigen retrieval conditions tested. Antibodies against CD4 (clone CT-4), TCRγδ, TCRαVβ1, CSF-1R, and MHC class II (clone 21‐1A6) were effective only in ZFPE tissue. In ducks, cells in both FFPE and ZFPE tissues were immunolabeled by five antibodies under some of the conditions tested. Antigen retrieval suitable for cellular membrane antigen tended to be heat for FFPE tissues and no treatment for ZFPE tissues. Heat-induced antigen retrieval allowed for better detection of nuclear antigens in both FFPE and ZFPE sections. Our results indicate that commercially available antibodies can immunohistochemically detect some of chicken and duck immune cell subsets in paraffin-embedded sections.
期刊介绍:
The journal reports basic, comparative and clinical immunology as they pertain to the animal species designated here: livestock, poultry, and fish species that are major food animals and companion animals such as cats, dogs, horses and camels, and wildlife species that act as reservoirs for food, companion or human infectious diseases, or as models for human disease.
Rodent models of infectious diseases that are of importance in the animal species indicated above,when the disease requires a level of containment that is not readily available for larger animal experimentation (ABSL3), will be considered. Papers on rabbits, lizards, guinea pigs, badgers, armadillos, elephants, antelope, and buffalo will be reviewed if the research advances our fundamental understanding of immunology, or if they act as a reservoir of infectious disease for the primary animal species designated above, or for humans. Manuscripts employing other species will be reviewed if justified as fitting into the categories above.
The following topics are appropriate: biology of cells and mechanisms of the immune system, immunochemistry, immunodeficiencies, immunodiagnosis, immunogenetics, immunopathology, immunology of infectious disease and tumors, immunoprophylaxis including vaccine development and delivery, immunological aspects of pregnancy including passive immunity, autoimmuity, neuroimmunology, and transplanatation immunology. Manuscripts that describe new genes and development of tools such as monoclonal antibodies are also of interest when part of a larger biological study. Studies employing extracts or constituents (plant extracts, feed additives or microbiome) must be sufficiently defined to be reproduced in other laboratories and also provide evidence for possible mechanisms and not simply show an effect on the immune system.