Immunohistochemical Markers for the Rodent Immune System

Department of Molecular Cell Biology and Immunology, VU University Amsterdam, Amsterdamo, North Holland, Netherlands
Toxicologic Pathology (Impact Factor: 2.14). 02/2006; 34(5):616-30. DOI: 10.1080/01926230600941340
Source: PubMed


The responses to insults including chemical toxins, irradiation and infectious agents involve morphologic, biochemical and molecular changes in the immune system. The changes in specific tissues and cells often can be detected by histopathology and its associated field of immunohistochemistry (IHC). Cells normally express specific proteins (antigens) that can be detected by IHC. When responses to xenobiotics occur, cells often up or down regulate proteins. The art of IHC requires specialized procedures for detection of antigens. Fixation, tissue processing, immunoreactions and antigen retrieval methods are important elements of IHC. We review the antibodies, their sources, use of frozen or fixed paraffin-embedded tissues and specific IHC methods including antigen retrieval and illustrate how they can be effectively used to characterize the immunotoxicologic effects of agents.

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    • "The primary antibodies were labeled with FITC-conjugated goat anti-rabbit IgG [H + L] (green, ZSGB-BIO) and TRITC-conjugated goat anti-mouse IgG [H + L] (red, Boster), Dylight™594 goat anti-Rabbit IgG [H + L], and Dylight™488 goat anti-Mouse IgG [H + L] (Muletiscience). The protocols were performed as in previous articles [18–20]. "
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    • "Some parts of lymph node specimens prepared by perfusion-fixation were infiltrated in PBS containing 30% sucrose [44], and 6 µm cryosections were cut. They were immunostained for LYVE-1 with the common immunoperoxidase-DAB method as reported before [40, 41]. "
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    • "Mice were observed for 8 to 12 months and necropsied when signs of splenomegaly, lymphadenopathy, labored breathing or lethargy were noted or the experiment was terminated at 12 months. Diagnosis was based on gross findings, microscopic examination of H&E stained formalin fixed, paraffin embedded tissues or studied by IHC using the anti-p30 antibody, anti-CD3 for T-cell lineage identification (DAKO Corporation, Carpinteria, CA Catalog # A452), and anti-PAX5 for B-cell lineage (Goat anti-Pax 5, Santa Cruz Biotechnology, Santa Cruz, CA, Catalog #sc-1974) [14]. Criteria for histopathological diagnosis were as described [15]. "
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