Detection and Characterization of Paraproteinemia in Canine Chronic B‐cell Lymphocytic Leukemia Using Routine and Free Light Chain Immunofixation

Abstract Background Hyperglobulinemia is reported in 26% of canine chronic B‐cell lymphocytic leukemia (B‐CLL) cases. However, few cases have been characterized by protein electrophoresis and immunofixation (IF), and the incidence of a monoclonal protein (M‐protein) is unknown using these techniques. Objective To characterize and determine the proportion of canine B‐CLL cases with an M‐protein using plasma protein electrophoresis (PPE), routine and free light chain (fLC) IF, and to assess if productive B‐CLL cases express MUM1/IRF4 by cell tube block (CTB). Methods PPE, routine (targeting IgG, IgA, IgM, IgG4, and light chain) and fLC IF were performed using 48 dog B‐CLL plasma samples from patients diagnosed via peripheral blood flow cytometry. CTB was performed on a separate cohort of 15 patients. Results Hyperproteinemia (>7.5 g/dL) was present in 17/48 cases (35%). An M‐protein was detected in 32/48 cases (67%). Of these, 19/32 cases (59%) had only complete (monoclonal heavy and light chain) M‐proteins detected, 10/32 cases (31%) had both complete and fLC M‐proteins detected, and 3/32 cases (9%) had only an fLC M‐protein detected. IgM was the most common clonal immunoglobulin isotype detected (23 cases). CD21+ cell counts were higher in cases with detectable M‐protein. Plasma fLC IF suggested β‐γ region interference, likely caused by clotting proteins. All B‐CLL cases consistently expressed PAX5 and did not express MUM1/IRF4. Conclusions Most B‐CLL cases had an M‐protein and were not hyperproteinemic. Most cases with paraproteins had a complete IgM monoclonal gammopathy; a subset had documented fLCs. The prognostic significance of heavy and fLC presence should be evaluated.

[1]  Paul R. Avery,et al.  Clinical outcome and prognostic factors in dogs with B‐cell chronic lymphocytic leukemia: A retrospective study , 2021, Journal of veterinary internal medicine.

[2]  Paul R. Avery,et al.  Diagnostic performance of routine electrophoresis and immunofixation for the detection of immunoglobulin paraproteins (M-Proteins) in dogs with multiple myeloma and related disorders: Part 2-Toward improved diagnostic performance. , 2021, Veterinary clinical pathology.

[3]  Paul R. Avery,et al.  Diagnostic performance of routine electrophoresis and immunofixation for the detection of immunoglobulin paraproteins (M-Proteins) in dogs with multiple myeloma and related disorders: Part 1 - Current performance. , 2021, Veterinary clinical pathology.

[4]  R. Harris,et al.  Light chain myeloma and detection of free light chains in serum and urine of dogs and cats , 2021, Journal of Veterinary Internal Medicine.

[5]  Paul R. Avery,et al.  Polyclonal B‐cell lymphocytosis in English bulldogs , 2020, Journal of veterinary internal medicine.

[6]  M. Varettoni,et al.  Monoclonal gammopathy and serum immunoglobulin levels as prognostic factors in chronic lymphocytic leukaemia , 2020, British journal of haematology.

[7]  A. R. Moore,et al.  Identification of canine IgG and its subclasses, IgG1, IgG2, IgG3 and IgG4, by immunofixation and commercially available antisera. , 2020, Veterinary immunology and immunopathology.

[8]  A. Avery,et al.  Validation and method comparison of the use of densitometry to quantify monoclonal proteins in canine sera. , 2019, Veterinary clinical pathology.

[9]  R. Marcos,et al.  Cell tube block: a new technique to produce cell blocks from fluid cytology samples , 2017, Veterinary clinical pathology.

[10]  P. Morley,et al.  Breed Distribution and Clinical Characteristics of B Cell Chronic Lymphocytic Leukemia in Dogs , 2016, Journal of veterinary internal medicine.

[11]  E. Guérin,et al.  IgM peak independently predicts treatment-free survival in chronic lymphocytic leukemia and correlates with accumulation of adverse oncogenetic events , 2014, Leukemia.

[12]  S. Harding,et al.  Prognostic Significance of Serum Free Light Chains in Chronic Lymphocytic Leukemia , 2013, Advances in hematology.

[13]  Jian-yong Li,et al.  Prognostic significance of serum immunoglobulin paraprotein in patients with chronic lymphocytic leukemia. , 2011, Leukemia research.

[14]  J. Cerhan,et al.  Monoclonal and polyclonal serum free light chains and clinical outcome in chronic lymphocytic leukemia. , 2010, Blood.

[15]  Michael Hallek,et al.  Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. , 2008, Blood.

[16]  Joseph A. DiGiuseppe,et al.  Laboratory, morphologic, and immunophenotypic correlates of surface immunoglobulin heavy chain isotype expression in B-cell chronic lymphocytic leukemia. , 2001, American journal of clinical pathology.

[17]  J. Ultmann,et al.  Chronic lymphocytic leukaemia and its relationship to other lymphoproliferative disorders. , 1977, Clinics in haematology.

[18]  Dailin Li,et al.  Candidate recommendations for protein electrophoresis reporting from the Canadian Society of Clinical Chemists Monoclonal Gammopathy Working Group. , 2018, Clinical biochemistry.

[19]  P. Néve,et al.  Surface membrane immunoglobulin expression in chronic lymphocytic leukemia. , 1994, Acta haematologica.

[20]  R. Matus,et al.  Chronic lymphocytic leukemia in the dog: 22 cases (1974-1984). , 1986, Journal of the American Veterinary Medical Association.