Roger Burnett

Immunohistochemistry for light microscopy in safety evaluation of therapeutic agents: an overview.

The history of immunohistochemistry started in 1941 when Coons identified pneumococci using a direct fluorescent method. Then followed the indirect method, the addition of horseradish peroxidase, the peroxidase anti-peroxidase technique of 1979 and the use of the Avidin and Biotin complex in the early 1980s. This sequence of events can help one appreciate the differences in these various techniques and their increased sophistication and sensitivity. The technique has been applied in the field of safety evaluation of new pharmaceutical products. Examples of current projects are used to illustrate the scope of the application. The use of an antibody to detect proliferating cell nuclear antigen has, in a pilot study with the popliteal lymph node assay, provided a method of differentiating an irritant response to acetone from an immune response to hydrazine. In hydrazine-treated rats the proliferation is mainly in the follicular region whilst it is mainly sinusoidal in animals treated with acetone. In the guinea-pig maximisation test, initial work with dinitrochlorobenzene suggests that detection of Langerhans cells may aid the differentiation of an irritant from an immune response. The preclinical assessment of antibodies intended for therapeutic use in man requires immunohistochemistry to be used to identify any human tissues which show a cross-reactivity. The major problems are not in the test itself but in obtaining suitable material. Identification of hormones is a useful tool for assessing the effects of releasing factors and has proved useful in aiding tumour identification in routine carcinogenicity studies. In a rare case, detection of prolactin in cell deposits in the lungs of a rat confirmed that this was a metastasis from a pituitary carcinoma. The application of immunohistochemical techniques to preclinical assessment of drugs should always be considered. In particular, it is recommended that appropriate samples should be conserved from routine studies in order to permit these techniques to be performed, if considered appropriate in the light of findings during routine histological examination.

Histopathological lesions following intramuscular administration of saline in laboratory rodents and rabbits

This review was undertaken to assess the nature and incidence of procedure-related changes in mice, rats and rabbits which received saline solution by intramuscular injection. Data were collected on the injection sites from 7 studies representing 152 animals. The original observations by the different study pathologists from both control and treated animals were evaluated in order to create a glossary of preferred terms to be used in toxicology studies. These standardized terms were then applied to changes observed in the saline-treated animals. The review showed that the most severe of the procedure-related lesions were only of a slight level. Two days post-injection, the local reactions were mainly composed of minimal infiltration by mononuclear cells (lymphocytes and macrophages) with occasional degeneration of myofibres. From 10 to 42 days post-injection, lesions showed regeneration of myofibres and some fibrosis. In rats, the number of injections at each site influenced inflammatory infiltrate and degenerative lesions.