Anti-Interleukin-10R1 Monoclonal Antibody Enhances Bacillus Calmette-Guérin Induced T-Helper Type 1 Immune Responses and Antitumor Immunity in a Mouse Orthotopic Model of Bladder Cancer

Bockholt, Nathan A.
Knudson, Matthew J.
Henning, Jonathan R.
Maymí, José L.
Weady, Peter
Smith, George J. III
Eisenbraun, Michael D.
Fraser, James D.
O'Donnell, Michael A.
Luo, Yi

Department of Urology, University of Iowa, Iowa City, Iowa, and Pfizer, Inc. (PW, GJS, MDE, JDF), San Diego, California

^‡Correspondence: Department of Urology, University of Iowa, 375 Newton Rd., 3202 MERF, Iowa City, Iowa 52242 (telephone: 319-335-9835; FAX: 319-384-4652)

E-mail: [email protected]

^*Financial interest and/or other relationship with Astellas and GlaxoSmithKline.

E-mail: [email protected]

^†Financial interest and/or other relationship with Pfizer.

E-mail: [email protected]

Submitted August 31, 2011.

Study received University of Iowa animal care and use committee approval.

Supported by Pfizer.

Journal of Urology 187(6):p 2228-2235, June 2012.

Purpose:

Proper induction of the T-helper type 1 immune response is required for effective bacillus Calmette-Guérin immunotherapy for bladder cancer. Interleukin-10 down-regulates the T-helper 1 response and is associated with bacillus Calmette-Guérin failure. We investigated whether blocking interleukin-10 receptor 1 would enhance the bacillus Calmette-Guérin induced T-helper type 1 immune response and anti-bladder cancer immunity in a mouse model.

Materials and Methods:

Splenocytes were incubated with bacillus Calmette-Guérin or bacillus Calmette-Guérin plus control IgG1, anti-interleukin-10 receptor 1 mAb or anti-interleukin-10 neutralizing mAb, followed by enzyme-linked immunosorbent assay of interferon-γ production. Bladder RNA was extracted after intravesical bacillus Calmette-Guérin plus intraperitoneal IgG1 or anti-interleukin-10 receptor 1 mAb and analyzed by reverse transcriptase and/or quantitative polymerase chain reaction. Urine was collected and analyzed by enzyme-linked immunosorbent assay. Mice bearing a luciferase expressing MB49 orthotopic tumor were treated with intravesical bacillus Calmette-Guérin plus intraperitoneal IgG1 or anti-interleukin-10 receptor 1 mAb. Tumor response was assessed by bioluminescent imaging and bladder weight measurement.

Results:

Bacillus Calmette-Guérin plus anti-interleukin-10R1 mAb induced significantly higher interferon-γ production by splenocytes than bacillus Calmette-Guérin plus anti-interleukin-10 mAb. Bacillus Calmette-Guérin plus anti-interleukin-10 receptor 1 mAb also induced significantly higher interferon-γ mRNA and protein in bladder and urine, respectively, in a dose dependent manner. Treatment with phosphate buffered saline, bacillus Calmette-Guérin plus control IgG1 and bacillus Calmette-Guérin plus anti-interleukin-10 receptor 1 mAb showed a 0% tumor-free rate with a 20% death rate, a 20% tumor-free rate with a 20% death rate and a 40% tumor-free rate with a 0% death rate, respectively. Bladder weight also revealed the effect of anti-interleukin-10 receptor 1 mAb on the bacillus Calmette-Guérin induced bladder tumor response.

Conclusions:

Anti-interleukin-10 receptor 1 mAb enhanced the bacillus Calmette-Guérin induced T-helper type 1 immune response and anti-bladder cancer immunity. A humanized form of this mAb warrants future investigation for bacillus Calmette-Guérin treatment of bladder cancer.

Abbreviations and Acronyms: BCG: bacillus Calmette-Guérin; ELISA: enzyme-linked immunosorbent assay; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; IFN: interferon; IL: interleukin; IL-10R: IL-10 receptor; IL-12p35: IL-12, p35 subunit; IP-10: IFN-inducible protein 10; Luc: luciferase; MCP-1: monocyte chemoattractant protein 1; MDC: macrophage-derived chemokine; OD: optical density; PBS: phosphate buffered saline; PMA: phorbol myristate acetate; qPCR: quantitative polymerase chain reaction; RT-PCR: reverse transcriptase-polymerase chain reaction; TH: T-helper type; TNF: tumor necrosis factor.