| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2000: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
(1) Developing a method for measuring the concentration of serum amyloid A (SAA) used as ainflammatory marker
We have successfully constructed a highly expressing recombinant feline (rf) SAA expression vector which expresses feline (f) SAA in Escherichia coli. We have also purified the rfSAA (reference paper 5), as it was difficult to obtain large amount of native SAA for our purpose. Subseqeuently, a panel of monoclonal antibodies specific for fSAA have been prepared by using the rfSAA (reference paper 1). Then an ELISA technique employing the monoclonal antibodies was developed and used in the estimation of fSAA.The results showed that fSAA was able to be used as an inflammatory marker, and is highly usefut for understanding the pathological state of inflammatory diseases (unpublished data).
(2) Usage of T-cell costimulatorymolecules and cytokines related to inflammatory diseases in dogs and cats.
Canine and feline T-cell costimulatory molecules, CTLA-4 and CD28 (reference paper 2, 3, 6
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), and a cytokine, IL-4 (reference paper 7), which are related to inflammatory diseases, such as autoimmune diseases, allergies etc, have been cloned and the proteins expressed (unpublished data). The application of these molecules both in reseach as well as for clinical purposes is in study.
(3) Development of a method for detecting anti neutrophil cytoplasmic antibody (ANCA) used as a vasculitis marker in small animals.
We have found that ANCA can be detected in various diseases of dogs and cats by using Indirect Fluorescent Antibody Test and ELISA (presentation 1, 2). The method for detecting ANCA is being developed, based on the previous results, as described in Section 2.
(4) Understanding of the relationship between T-cell costimulatory molecules, cytokines, acute phase proteins (SAA, C-reactive protein), and ANCA.
The dynamic changes of acute phase proteins in diseased dogs and cats have been analyzed, which is an important indicator of disease status (reference paper 4 and 8 unpulished). In future, the relationship between the above mentioned molucules will be described, from the results of section 1, 2 and 3, which will be highly useful in clinical applications. Less
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