Masters Theses

Date of Award

5-1995

Degree Type

Thesis

Degree Name

Master of Science

Major

Life Sciences

Major Professor

Stephen J. Kennel

Committee Members

Peggy Terzaghi-Howe, John Koontz, Carl Wust

Abstract

Renal cell carcinoma is responsible for 2% of all malignant tumors in adults and is one of the most challenging tumors to treat by the clinical oncologist. Unless early primary lesions (stage I or stage II) are identified and completely removed, cures are uncommon. Patients presenting with late stage or metastatic disease have a relentless progression of disease and a short survival. The exact etiology of renal cell carcinoma is poorly understood and effective measures of prevention have not been determined. Current therapeutic methodologies used in the treatment of late stage and metastatic renal cell carcinoma have been generally ineffective, failing to realize a prolonged control of tumor spread. A monoclonal antibody, 469-5A, has been generated using human renal cell carcinoma lysate as an immunogen in BALB/c mice. The monoclonal antibody recognizes a glycoprotein, designated gp 45/55, which is overexpressed in 18/18 renal cell carcinomas observed. Evidence from Western blot, including staining pattern, molecular weight range, and increased detectability after neuraminidase treatment demonstrated that monoclonal antibody 5A recognized a similar glycoprotein in both normal kidney and tumor tissues. However, Western blot data indicates that gp 45/55 expression is 2 to 6-fold greater in RCC than normal kidney tissues. Furthermore, treatment with various combinations of other glycosidases caused shifts in the detected molecular weight in Western blot, indicating that the 5A epitope was probably protein in nature. Immunohistochemistry supported Western blot data, demonstrating an increase in monoclonal antibody 5A staining in tumor tissue compared to normal kidney tissue. Monoclonal antibody 5A staining was restricted to proximal tubule epithelium in normal kidney but was detected on most of the tumor epithelial cells. Expression of 5A antigen was also detected by Western blot and/or immunohistochemistry in other normal human tissues as well as in lymphoma tissues. Based on the same identification criteria, the 5A monoclonal antibody detected a similar molecule, gp 45/55, in Western blot of other normal tissues as had been detected in normal kidney and renal cell carcinoma. However, distribution data comparing results from immunohistochemistry and from Western blot of various normal and tumor tissues demonstrated that gp 45/55 was not detected in IHC of small intestine and spleen but was detected in Western blot. The results presented demonstrate the overexpression of detectable gp 45/55 in renal carcinoma cells. It is also shown that the probable epitope for the 5A monoclonal antibody is protein in nature. Western blot data indicates that monoclonal antibody 5A recognizes the same antigen, gp 45/55, in normal kidney and tumor cells. Similarly, gp 45/55 is expressed in cells of other normal tissues as well as in lymphoma tissues. The potential of gp 45/55 as a renal carcinoma marker and of the immunodiagnostic and immunotherapeutic use of monoclonal antibody 5A is discussed.

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