Targeting of renal tumors with mAb 138H11 against human gamma-glutamyltransferase in a novel syngeneic mouse model and cloning of recombinant 138H11 derivatives

Continuing our strategy for a targeted therapy of metastatic renal cell carcinoma (RCC), a tumor which resists chemo- or radiotherapy, we now determined the primary structure of the antigen binding region of the monoclonal antibody 138H11. MAb 138H11, produced against human renal gamma-glutamyltransferase (GGT), stained over 99% clear cell and papillary RCC as well as other carcinomas on frozen sections, showing a membranous expression of the target antigen. In contrast, in normal kidneys GGT is restricted to the brush-border in the lumen of proximal tubules and thus not accessible to the circulation. Human tumor-bearing kidneys perfused in an
extra-corporeal system with 99mTc-138H11 revealed a high, specific uptake into the tumor. In vivo, a 138H11-Calicheamicin theta conjugate was very effective in reducing the tumor size and preventing or significantly delaying the regrowth of residual tumor cells in xenografted nude mice (Cancer Res. 2000, 60, 6089-6094). However, these nude mice are not useful for evaluating ADCC of naked 138H11 or immunoactivating conjugates. For creating a syngenic, immunocompetent mouse model bearing a tumor sensitive to mAb 138H11, we stably transfected the murine RCC RENCA cell line with the human GGT-gene. FACS-analysis revealed that transfected cells were positive for mAb 138H11, in contrast to wild type cells. The transfected cells were growing s.c. in
Balb/c mice without signs of rejection by the host. The mice showed a higher uptake of 99m-Tc-labelled 138H11 into the GGT positive RENCA tumors compared to the wild type tumors. This new model will be usful in the future for evaluating therapeutic effects of potential constructs made with the newly cloned recombinant 138H11-derivatives such as stabilized scFv, diabodies and triabodies.