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Clinical and vaccine immunology : CVI2015; 22(11); 1133-1145; doi: 10.1128/CVI.00374-15

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.
Copyright © 2015, American Society for Microbiology. All Rights Reserved.
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Publication Date: 2015-08-26 PubMed ID: 26311245PubMed Central: PMC4622111DOI: 10.1128/CVI.00374-15Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article focuses on the study and classification of B cell diseases in horses using protein and molecular methods. The main goal was to better understand these diseases by analysing their gene and protein expressions, mutations, and other bio-markers.

Research Objective

  • The study was designed to use molecular techniques to further delve into the classification of B cell lymphoproliferative diseases found in horses. Especially, it focused on five adult horses which were experiencing a rare condition of monoclonal gammopathy, B cell leukemia, and simultaneous lymphadenopathy (lymphoma/leukemia).

Methods and Techniques

  • The researchers drew from various scientific processes and techniques, including gene and cell surface molecule expression, Ig isotype concentrations, IGHV region domain sequencing, and spectratyping to phenotypically characterize the B cell neoplasms.
  • An in-depth investigation of the B cell-associated developmental genes EBF1, PAX5, and CD19, along with the plasma cell-associated marker CD38, was also conducted by measuring their mRNA expression levels.
  • Further, the dominant monoclonal peak was identified with Ig spectratyping and secreted isotype concentration.
  • IGHV domain of the leukemic cells was sequence analyzed to identify mutated Immunoglobulins.

Findings

  • It was observed that all the five horsed studied had hyperglobulinemia due to IgG1 or IgG4/7 monoclonal gammopathy
  • A high number of IgG1- or IgG4/7-positive cells was found in leukocyte immunophenotyping of peripheral blood.
  • Most leukemic cells were deprived of the surface B cell markers, CD19 and CD21
  • The gene expression of IGHG1 or IGHG4/7 was aligned with the surface protein expression
  • The mRNA expression levels of EBF1, PAX5, and CD19 genes were high when compared to that of CD38 gene.
  • The sequence analysis revealed mutations in the IGHV domain of the leukemic cells.

Conclusion

  • The study concluded that the molecular and protein techniques used could identify neoplastic cells, suggesting a developmental transition from B cell and plasma cell stages.
  • These techniques can be effective in classifying equine B cell lymphoproliferative diseases.

Cite This Article

APA
Badial PR, Tallmadge RL, Miller S, Stokol T, Richards K, Borges AS, Felippe MJ. (2015). Applied Protein and Molecular Techniques for Characterization of B Cell Neoplasms in Horses. Clin Vaccine Immunol, 22(11), 1133-1145. https://doi.org/10.1128/CVI.00374-15

Publication

Clinical and vaccine immunology : CVI
ISSN: 1556-679X
NlmUniqueID: 101252125
Country: United States
Language: English
Volume: 22
Issue: 11
Pages: 1133-1145

Researcher Affiliations

Badial, Peres R
  • 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.
Tallmadge, Rebecca L
  • Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA.
Miller, Steven
  • Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA.
Stokol, Tracy
  • Department of Population Medicine and Diagnostic Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA.
Richards, Kristy
  • Department of Biomedical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA.
Borges, Alexandre S
  • Department of Veterinary Clinical Sciences, School of Veterinary Medicine and Animal Sciences, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil.
Felippe, M Julia B
  • Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Ithaca, New York, USA mbf6@cornell.edu.

MeSH Terms

  • Animals
  • Antigens, CD19 / analysis
  • B-Lymphocytes / immunology
  • B-Lymphocytes / metabolism
  • B-Lymphocytes / pathology
  • Horse Diseases / genetics
  • Horses
  • Immunoglobulin Heavy Chains / genetics
  • Immunophenotyping
  • Leukemia, B-Cell / genetics
  • Leukemia, B-Cell / immunology
  • Leukemia, B-Cell / veterinary
  • Lymphatic Diseases / genetics
  • Lymphatic Diseases / immunology
  • Lymphatic Diseases / veterinary
  • Lymphopenia / genetics
  • Lymphopenia / immunology
  • Lymphopenia / veterinary
  • Lymphoproliferative Disorders / classification
  • Lymphoproliferative Disorders / genetics
  • Lymphoproliferative Disorders / immunology
  • Lymphoproliferative Disorders / veterinary
  • PAX5 Transcription Factor / analysis
  • Paraproteinemias / genetics
  • Paraproteinemias / immunology
  • Paraproteinemias / veterinary
  • Plasma Cells
  • Receptors, Complement 3d / analysis
  • T-Lymphocytes

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