title: Applied Protein and Molecular Techniques for Characterization of B Cell Neoplasms in Horses.
abstract: Mature B cell neoplasms cover a spectrum of diseases involving lymphoid tissues (lymphoma) or blood (leukemia), with an overlap between these two presentations. Previous studies describing equine lymphoid neoplasias have not included analyses of clonality using molecular techniques. The objective of this study was to use molecular techniques to advance the classification of B cell lymphoproliferative diseases in five adult equine patients with a rare condition of monoclonal gammopathy, B cell leukemia, and concurrent lymphadenopathy (lymphoma/leukemia). The B cell neoplasms were phenotypically characterized by gene and cell surface molecule expression, secreted immunoglobulin (Ig) isotype concentrations, Ig heavy-chain variable (IGHV) region domain sequencing, and spectratyping. All five patients had hyperglobulinemia due to IgG1 or IgG4/7 monoclonal gammopathy. Peripheral blood leukocyte immunophenotyping revealed high proportions of IgG1- or IgG4/7-positive cells and relative T cell lymphopenia. Most leukemic cells lacked the surface B cell markers CD19 and CD21. IGHG1 or IGHG4/7 gene expression was consistent with surface protein expression, and secreted isotype and Ig spectratyping revealed one dominant monoclonal peak. The mRNA expression of the B cell-associated developmental genes EBF1, PAX5, and CD19 was high compared to that of the plasma cell-associated marker CD38. Sequence analysis of the IGHV domain of leukemic cells revealed mutated Igs. In conclusion, the protein and molecular techniques used in this study identified neoplastic cells compatible with a developmental transition between B cell and plasma cell stages, and they can be used for the classification of equine B cell lymphoproliferative disease.
authors: Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA Department of Veterinary Clinical Sciences, School of Veterinary Medicine and Animal Sciences, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil.
authors: Peres R
authors: PR
authors: Badial
authors: Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA.
authors: Rebecca L
authors: RL
authors: Tallmadge
authors: Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA.
authors: Steven
authors: S
authors: Miller
authors: Department of Population Medicine and Diagnostic Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA.
authors: Tracy
authors: T
authors: Stokol
authors: Department of Biomedical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA.
authors: Kristy
authors: K
authors: Richards
authors: Department of Veterinary Clinical Sciences, School of Veterinary Medicine and Animal Sciences, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil.
authors: Alexandre S
authors: AS
authors: Borges
authors: Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA mbf6@cornell.edu.
authors: M Julia B
authors: MJ
authors: Felippe
mesh_terms: D000818:Animals; D018941:Antigens, CD19 / Q000032:analysis; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D006734:Horse Diseases / Q000235:genetics*; D006736:Horses; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D016130:Immunophenotyping; D015448:Leukemia, B-Cell / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D008206:Lymphatic Diseases / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D008231:Lymphopenia / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D008232:Lymphoproliferative Disorders / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D051757:PAX5 Transcription Factor / Q000032:analysis; D010265:Paraproteinemias / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D010950:Plasma Cells; D017464:Receptors, Complement 3d / Q000032:analysis; D013601:T-Lymphocytes
chemical_list: D018941:Antigens, CD19; D007143:Immunoglobulin Heavy Chains; D051757:PAX5 Transcription Factor; D017464:Receptors, Complement 3d