(Immunohistochemistry)

By

( ‫ )ساندي عادل اسحاق‬Sage-3

PRINCIPLES OF IMMUNOHISTOCHEMISTRY (IHC).

The selection of antibodies for the immunohistochemically testing is made

on the basis of their tumor specificity and the likelihood that they will react
with the tumor under evaluation. After tissue sections are incubated with
the prospective antibodies, positive reactions (tumor antigen- antibody
binding)
are identified through the application of one of several detection systems.
Those that have the greatest sensitivity use a secondary antibody, reactive
against the primary antibody, which is conjugated or linked to an enzyme
marker. This system tends to be very sensitive because it allows for the
attachment of a relatively large number of enzyme molecules, such as
peroxidise, at the antigen site. The color of the reaction is determined by
the selection of a precipitating chromogen, usually diaminobenzidine
(brown)or aminoethylcarbazole (red), with which the enzyme
Applications
Since IHC involves specific antigen–antibody reactions, it has apparent
advantage over traditionally used special enzyme staining techniques that
identify only a limited number of proteins, enzymes, and tissue structures.
Therefore, IHC has become a crucial technique and is widely used in many
medical research laboratories as well as clinical diagnostics . Prognostic
Markers in Cancer To predict the prognosis of tumors by identification of
enzymes, tumor-specific antigens, oncogenes, tumor suppressor genes, and
tumor cell proliferation markers. Analysis of tumors by these methods is a
significant improvement over the conventional prognostic considerations by
clinical staging and histologic grading. IHC is used for disease diagnosis, drug
development, and biological research. Using specific tumor markers,
physicians use IHC to diagnose a cancer as benign or malignant, determine the
stage and grade of a tumor, and identify the cell type and origin of a metastasis
to find the site of the primary tumor. IHC is also used in drug development to
test drug efficacy by detecting either the activity or the up- or down-regulation
of disease targets.
Prediction of Response to Therapy
IHC is widely used to predict therapeutic response in two important tumors,
i.e. carcinoma of breast and prostate. Both these tumors are under the growth
regulation of the hormones estrogen and androgen, respectively. The specific
receptors for these growth regulating hormones are located on respective tumor
cells. Tumors expressing high level of receptor positivity would respond
favorably to removal of the endogenous source of such hormones or hormonal
therapy is administered to lower their levels – estrogen therapy in prostate
cancer and androgen therapy in breast cancer. Neurodegenerative Disorders
Degenerative disorders of the nervous system include a wide range of diseases
characterized by the dysfunction and death of specific, selectively vulnerable
populations of nerve cells. It has played an increasingly important role in the
subclassification of neurodegenerative disorders and the development of
consensus criteria for their diagnosis.