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In vitro Canine Distemper Virus Infection of Canine Lymphoid Cells: A Prelude to Oncolytic Therapy for Lymphoma

Departments of ¹ Surgical and Radiological Sciences and ² Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, California; ³ INRS-Institut Armand-Frappier, Université du Québec, Quebec, Canada; and ⁴ Molecular Medicine Program, Mayo Clinic, Rochester, Minnesota

Requests for reprints: Cheryl A. London, Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, 2112 Tupper Hall, One Shields Avenue, Davis, CA 95616. Phone: 530-754-5011; Fax: 530-754-4918; E-mail: calondon{at}ucdavis.edu.

Purpose: Measles virus (MV) causes the regression of human lymphoma xenografts. The purpose of this study was to determine if canine lymphoid cells could be infected in vitro with MV or canine distemper virus (CDV, the canine Morbillivirus equivalent of MV) and determine if in vitro viral infection leads to apoptotic cell death.

Experimental Design: Reverse transcriptase-PCR was used to examine the expression of both signal lymphocyte activation molecule (CD150) and membrane cofactor molecule (CD46) mRNA. An attenuated CDV expressing enhanced green fluorescent protein was used to infect canine cells in vitro. Both flow cytometry and reverse transcriptase-PCR was used to document CDV infection. Cell death was examined using a propidium iodide staining assay and Annexin V binding.

Results: Canine lymphoid cell lines and neoplastic B and T lymphocytes collected from dogs with spontaneous lymphoma expressed the Morbillivirus receptor CD150 mRNA. In contrast, only neoplastic lymphocytes expressed detectable levels of CD46 mRNA. Although MV did not infect canine cells, CDV efficiently infected between 40% and 70% of all three canine lymphoid lines tested. More importantly, CDV infected 50% to 90% of neoplastic lymphocytes isolated from dogs with both B and T cell lymphoma. Apoptosis of CDV-infected cell lines was documented.

Conclusions: Attenuated CDV may be a useful treatment for canine lymphoma. As such, dogs with lymphoma may represent a biologically relevant large animal model to investigate the feasibility, safety, and efficacy of Morbillivirus therapy in a clinical setting with findings that may have direct applicability in the treatment of human non-Hodgkin's lymphoma.

Key Words: oncolysis • Morbillivirus • non-Hodgkin's lymphoma

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