Generation of Kidney Cancer-Specific Antitumor Immune Responses Using Peripheral Blood Monocytes Transduced With a Recombinant Adenovirus Encoding Carbonic Anhydrase 9

Abstract

Purpose: Carbonic anhydrase 9 (CA9) is the most promising molecular marker described for renal cell carcinoma (RCC) to date. We investigated whether transduction of monocytes from peripheral blood with adenovirus encoding the CA9 gene (AdV-CA9) could stimulate a T-cell mediated immune response against cancer cells expressing CA9. The ability to consistently generate a T-cell response is an important step toward the development of a CA9-specific RCC vaccine.Experimental Design: AdV-CA9 was generated using the AdEasy system. AdV-CA9-transduced peripheral blood mononuclear cell (PBMC)-derived monocytes were used to raise CTLs from autologous peripheral blood lymphocytes (PBLs). The ability of CTLs to lyse targets expressing CA9 was assessed by 51 Cr-release. Results: Monocytes were efficiently transduced with AdV-CA9. In five of six experiments, AdV-CA9-transduced monocytes were able to induce a population of CTLs from bulk PBLs. CTLs were capable of lysing autologous, but not allogeneic monocytes expressing CA9. Furthermore, CTLs were able to lyse autologous RCC tumor cells expressing CA9. The ability of CTLs to lyse relevant targets was blocked by anti-CD3, anti-CD8, and anti-MHC class I antibodies demonstrating a MHC class I restricted response.Conclusions: These results suggest that PBMC-derived monocytes transduced with AdV-CA9 can generate RCCspecific MHC class I restricted CTLs capable of lysing CA9-expressing cancer cells. Transduction of PBMCderived monocytes with adenovirus provides a simple and effective alternative to the use of dendritic cells for the induction of antigen-specific CTL.

Ontogeny and Oncogenesis Balance the Transcriptional Profile of Renal Cell Cancer

“…Although the highest level of CA II expression was noted in chromophobe RCCs, several clear cell RCCs expressed this gene, suggesting that, at least at the RNA level, there is overlap between the two types of RCCs (8). Interestingly, CA IX expression was specific for RCCs compared with normal renal tissue, confirming that this gene may be a better target for drug or immune therapy (32). Interestingly, the expression of CA IX was shared by other primary epithelial cancers but not melanoma metastases, suggesting that the expression of this gene is at least partly associated with oncogenesis and that it may not be solely responsible for the immune responsiveness of these diseases to immune therapy.…”

Molecular and cytogenetic insights into the pathogenesis, classification, differential diagnosis, and prognosis of renal epithelial neoplasms

“…CAIX expression correlates with tumor stage, grade, lymph node involvement, distant metastasis, survival, and response to immunotherapy [158,159]. Overexpression of CAIX has been seen in 70% to 96% of clear cell renal cell carcinomas [160,161]. Followup study of 63 cases of clear cell renal cell carcinoma showed that metastasis-free survival rate correlates with intensity of CAIX expression and is significantly better for patients whose tumors highly express CAIX as compared with patients whose tumors express it less strongly [157].…”

Biomarkers in renal cell carcinoma

“…In most cancers, CAIX is a marker for hypoxia with expression increased in bulky tumours and greatest expression in tumour cells immediately adjacent to areas of necrosis [6]. However, in clear cell RCC (ccRCC), CAIX expression is overexpressed due to inactivation of the VHL gene product and as a result overexpression of HIF-1a, a transcription factor for CAIX [7].In ccRCC, CAIX is an ideal biomarker that establishes diagnosis [8-10], determines prognosis [11][12][13][14], predicts treatment response [15,16] and serves as a target for therapy [17][18][19]. CAIX is a good biomarker for RCC (detected in 86% of RCC samples and only 9% of normal kidney [20], and present in more than 80% of primary and metastatic RCC [21]), and ccRCC in particular (94-100% of ccRCC stained positively for CAIX [12,21], whereas staining was absent in benign tumours such as oncocytoma [12]) (Fig.…”