A growing body of evidence suggests that immunotoxicity provides a considerable contribution to


onset and development of various disorders, including cancer and autoimmune diseases.137-139 Nevertheless, it was not until recently that this relatively new field of toxicology emerged as an important interface between the fields of novel drug design and pharmacology. Recognition of immunosuppressive properties of new pharmaceuticals during early drug development phase is very important to eliminate potentially dangerous substances from the drug pipelines. For example, treatment of patients diagnosed with Crohn's disease and rheumatoid arthritis with inflix-imab and etanercept (both drugs represent neutralizing anti-TNF antibodies) resulted in increased incidence of tuberculosis and histoplasmosis.140-143 Although these data did not result in withdrawal of any of the products from the pharmaceutical market, they helped initiate the strategy of preparing patients for anti-TNF therapy by screening for, and treatment of, latent tuberculosis prior to administration of anti-TNF medications. Immunosuppression caused by pharmaceuticals can also lead to the development of lymphomas and acute leukemia.144-146 Undesirable immunostimulation caused by pharmacological intervention include immunogenicity, hypersensitivity, and increased risk of autoimmune response. The standard toxicology endpoints employed for safety assessment of new pharmaceuticals primarily rely on clinical chemistry and histopathological evaluation of immune organs and were developed several decades ago. Currently, there is an increasing demand for the development of new methods for immunotoxicity assessment because of drug candidates' more complex structure as well as the application of new technologies in their manufacturing. The introduction of new molecular and immune cell biology methods into the immunotoxicology assessment framework is not a trivial and straightforward process. It requires not only scrupulous validation and standardization of the new techniques but also demonstration of the physiological relevance for the proposed battery of assays. These processes are expensive, time-consuming, and necessitate cooperation across the various pharmaceutical industry players. Unlike traditional drugs, multifunctional nanomaterials combine both chemistry-based and biotechnology-derived components, and therefore their characterization using standard methodologies requires adjustments and/or modification of classical experimental protocols. Below we will attempt to summarize data on critical aspects of immunotoxicological evaluation of nanomaterials and examine challenges in the application of standard methodologies for the assessments of nanoparticle safety to the immune system.

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