Multiple myeloma in dogs: Use of the cell block technique as a new diagnostic tool.

BACKGROUND: The diagnosis of multiple myeloma (MM) in dogs may be challenging and complex. The cell blocks are a diagnostic technique that allows the characterization of neoplastic cells and, therefore, might help in the diagnosis of atypical MM. OBJECTIVE: The objective of the present work is to describe three clinical cases in which the cell blocks and immunohistochemistry contributed to the definitive diagnosis of canine MM. METHODS: Three dogs, one female and two males, with different clinical signs, were presented for consultation with anemia, hyperproteinemia with monoclonal gammopathy, and the presence of plasmacytosis in the bone marrow. Cytologic analysis of the spleen was performed in two dogs and was suggestive of the presence of lymphocytes or plasma cells of a neoplastic nature in one of the cases and plasma cell hyperplasia associated with extramedullary hematopoiesis in the other. Given the hypotheses of lymphoid neoplasms with a plasma cell phenotype, cell blocks from aspiration punctures were performed for immunohistochemical analysis with anti-CD3, CD20, CD79αcy, PAX5, and MUM1 antibodies. RESULTS: The results revealed positive staining for MUM1 in 80% of the cells in the spleen cell block and for CD20 and MUM1 in 70% of the cells in the bone marrow cell blocks, with negative staining for the other antibodies. The immunophenotyping results allowed the diagnosis of MM in the three cases and excluded other lymphoid neoplasms. CONCLUSIONS: This work reinforces the importance of using cell blocks in the diagnosis of neoplasms by demonstrating their potential to aid the diagnosis of MM.

Cyclical 10-day dosing of melphalan for canine multiple myeloma.

Canine multiple myeloma (MM) is typically treated with melphalan chemotherapy. A protocol with repeated 10-day cyclical dosing of melphalan has been used at our institution but has not been described in the literature. Our objectives were to describe the outcome and adverse events of this protocol in a retrospective case series. We hypothesised the cyclical 10-day protocol would have similar outcomes compared to other reported chemotherapy protocols. Dogs diagnosed with MM that received melphalan treatment at Cornell University Hospital for Animals were identified through a database search. Records were retrospectively reviewed. Seventeen dogs met inclusion criteria. Lethargy was the most common presenting complaint. The median duration of clinical signs was 53 days (range, 2-150 days). Seventeen dogs had hyperglobulinemia with 16/17 having monoclonal gammopathies. Sixteen dogs had bone marrow aspiration and cytology performed at initial diagnosis and plasmacytosis was diagnosed in all. Based on serum globulin concentrations, 10 of 17 dogs (59%) achieved complete response (CR), and 3 dogs (18%) achieved partial response (PR), for an overall response rate of 76%. The median overall survival time was 512 days (range, 39-1065). Retinal detachment (n = 3) and maximum response of CR/PR (n = 13) were associated with overall survival on multivariate analysis (p = .045 and .046, respectively). Adverse events were minimal with diarrhoea being the most reported (n = 6). This cyclical 10-day protocol was better-tolerated with fewer adverse events than with other reported chemotherapy protocols, but response rate was also lower, likely due to a lower dosing intensity.

Multiple myeloma presenting as blepharitis in a horse.

Myeloma-related disorders, including multiple myeloma, extramedullary plasmacytoma, and solid osseous plasmacytoma, are rare in horses. Clinical complaints for myeloma-related disorders are nonspecific, and when present, M-protein location is more variable on protein electrophoresis in horses relative to dogs and cats. Here, we describe a case of a 15-year-old Thoroughbred mare who presented with recurrent blepharitis. Marked hyperglobulinemia was an incidental finding on routine hematologic and biochemical testing. Bone marrow aspiration consisted of >30% plasma cells, and serum protein electrophoresis demonstrated a monoclonal gammopathy in the alpha 2 fraction leading to a diagnosis of multiple myeloma. Immunofixation and radial immunodiffusion confirmed the presence of an IgG M-protein. Based on a restricted peak in the alpha 2 location, the specific M-protein is suspected to be IgG(T), an IgG isotype unique to horses. M-protein migration in horses is variable relative to dogs and cats, yet immunofixation can still be used to identify equine IgG M-protein isotypes. The unique clinical presentation in this case also serves as a reminder to consider neoplasia in horses with unusual or nonspecific clinical signs.

A retrospective study of canine oral extramedullary plasmacytoma over a 15-year period (July 2004-July 2019): Treatment, histologic parameters and clinical outcomes.

A total of 45 cases of canine oral extramedullary plasmacytomas (EMPs) presented to a tertiary referral institution over a 15-year period were examined. Histologic sections of 33 of these cases were examined for histopathologic prognostic indicators. Patients underwent variable treatment including surgical intervention, chemotherapy and/or radiation therapy. Long term survival was observed in the majority of dogs with a median survival time of 973 days (2-4315 days). However, almost 1/3 of dogs had progression of plasma cell disease, including two cases with myeloma-like progression. Histologic characterization of these tumours did not reveal criteria to predict tumour malignancy. However, cases without tumour progression did not exceed 28 mitotic figures in ten 400× fields (2.37 mm2 ). All cases with tumour related death showed at least moderate nuclear atypia. Oral EMPs may represent a local manifestation of systemic plasma cell disease or singular focal neoplasia.

Computed tomographic findings in dogs with multiple myeloma.

BACKGROUND: Computed tomography (CT) is considered the first-line imaging modality for human patients with suspected multiple myeloma (MM). Recently the diagnostic criteria for human MM have been updated. OBJECTIVES: To describe and provide a baseline of the CT features and distribution of osseous lesions in dogs diagnosed with MM and to describe the change of initial osseous lesions after the start of treatment in a subset of dogs. METHODS: Single-centre, retrospective, descriptive, case series. Dogs were included when they met the updated MM criteria and if a staging CT was performed at the time of diagnosis and prior to initiation of treatment. When available, change of osseous lesions was assessed on serial imaging studies. RESULTS: Thirteen dogs met the inclusion criteria. All dogs had involvement of the axial skeleton and 9/13 (69%) had concurrent involvement of the appendicular skeleton. Large (≥1 cm), lytic, 'punched out' or expansile bony lesions and regions of permeative lysis were most common and mainly affected the vertebral column. Discrete intramedullary soft tissue attenuating lesions of the proximal appendicular skeleton were observed in 8/13 dogs (61%) and bilateral involvement of both humeri and femurs was seen in 4/9 dogs (44%). A subset of dogs underwent serial imaging and progressive replacement of the contrast-enhancing lesions with fat attenuating tissue was observed for all dogs, corresponding with clinical improvement. CONCLUSIONS: This case series provides a baseline knowledge of the initial and follow-up CT features in dogs diagnosed with MM based on updated criteria.

Diagnosing Multiple Myeloma and Related Disorders.

This review provides current information on myeloma-related disorders, a group of plasma cell or immunoglobulin (Ig) secreting neoplasms including multiple myeloma, extramedullary plasmacytoma (both cutaneous and noncutaneous variants), solitary osseous plasmacytoma, Waldenström macroglobulinemia/lymphoplasmacytic lymphoma, Ig-secretory B-cell lymphoma, plasma cell leukemia, and monoclonal gammopathy of undetermined significance. The diagnostic procedures commonly used to characterize myeloma-related disorders, including cytopathology, histopathology, polymerase chain reaction for antigen receptor rearrangement, flow cytometry, and electrophoretic techniques are outlined and discussed.

Multiple Myeloma with Aberrant CD3 Expression in a Red-Lored Amazon Parrot (Amazona autumnalis).

A 20-year-old, female, red-lored Amazon parrot (Amazona autumnalis) was presented for a 2-week history of weakness. On physical examination, the bird was quiet, fluffed, weak, and had a distended coelom. Radiographic and ultrasound imaging revealed coelomic distention, increased pulmonary parenchymal opacity, renomegaly, dilated intestines, and a thickened ventricular wall. The results of a complete blood cell count indicated the patient was anemic (28%) and had intermediate to large lymphocytes with immature chromatin that were suspected to be neoplastic. Immunocytochemistry on peripheral blood determined that the suspected circulating neoplastic cells were cluster of differentiation (CD) 3+ and occasionally expressed multiple myeloma oncogene 1 (MUM1). Abnormalities from a plasma biochemistry panel were moderate hyperphosphatemia (6.8 mg/dL), marked hyperproteinemia (13.6 g/L), analbuminemia (0 g/dL), and marked hyperglobulinemia (13.6 g/dL). Agarose gel plasma protein electrophoresis documented the presence of albumin (1.2 g/dL) and monoclonal bands which, on reduced lithium dodecyl sulfate polyacrylamide gel electrophoresis, resolved as 60-kd and ∼25-kd bands consistent with immunoglobulin Y heavy and light chains. On the basis of these findings, multiple myeloma was diagnosed. Because of a poor prognosis, the bird was euthanized for postmortem examination. Bone marrow cytology from samples collected during the postmortem examination revealed 17.4% plasma cells and 24% large immature cells with occasional plasmacytoid features. Histopathologic findings included aggregates of neoplastic plasma cells in the bone marrow, spleen, kidney, liver, gastrointestinal tract, muscle, ovary, and brain. The neoplastic cells were strongly immunoreactive for MUM1 and cluster of differentiation 3 (CD3), but negative for CD79a, paired box protein 5, and CD20. This confirmed the clinical diagnosis of multiple myeloma. This report describes an avian immunoglobulin Y-secreting multiple myeloma with aberrant CD3 expression and pseudoanalbuminemia. Aberrant CD3 expression by avian multiple myeloma may explain previously published cases of birds with a monoclonal gammopathy and apparent T-cell lymphoma diagnosed by CD3 immunoreactivity.

A feline case of multiple myeloma treated with bortezomib.

BACKGROUND: Multiple myeloma (MM) is an uncommon neoplasm in cats. There is no established standard of treatment due to the rare occurrence of this disease in cats. Bortezomib is a proteasome inhibitor that serves as the first-line drug for MM in humans, but its effectiveness currently is unknown in feline MM. We present here the case report of a feline MM that exhibited a favorable response to bortezomib. CASE PRESENTATION: The case was an 11-year-old non-castrated male domestic cat with light-chain MM presenting with clinical symptoms (anorexia, fatigue, and vomiting), mild azotemia, and pancytopenia. The cat failed on melphalan with prednisolone (MP), so bortezomib (Velcade) was initiated on Day 88. A total of 6 cycles of the treatment was performed, with each treatment cycle consisting of twice-weekly subcutaneous administration for 2 weeks followed by a 1-week rest. The dose of bortezomib was 0.7 mg/m2 for first week and 1.0 mg/m2 for second week in the first cycle. A dose of 0.7 mg/m2 was used for subsequent cycles. Prednisolone was used concomitantly in the first 2 cycles. Following treatment with bortezomib, clinical symptoms disappeared and a decrease in serum globulin and recovery of pancytopenia were noted. A monoclonal gammopathy, overproduction of serum immunoglobulin light chain, and Bence-Jones proteinuria that existed at diagnosis were undetectable on Day 123. A monoclonal gammopathy also was not detectable at the end of the bortezomib treatment (Day 213). Anorexia, fatigue, and marked bone marrow toxicity were experienced when bortezomib was administrated at a dose of 1.0 mg/m2, while no recognizable toxicity was observed at a dose of 0.7 mg/m2 throughout the treatment period. The case was placed on follow-up and there was no evidence of relapse as of Day 243. CONCLUSIONS: Bortezomib was effective and durable for the treatment of this case of feline MM after failure with MP. Bortezomib was well-tolerated in this cat at a dose of 0.7 mg/m2, but not at 1.0 mg/m2. Bortezomib appears to be a drug worthy of further study for the treatment of feline MM.

Spurious laboratory results associated with immunoglobulin M gammopathy in a dog with multiple myeloma.

An 11 year old female-neutered Labrador presented for facial swelling. Clinicopathological abnormalities included hyperglobulinemia, azotemia, hypercalcemia, nonregenerative anemia, thrombocytopenia, and spurious hypoglycemia. Normoglycemia was subsequently confirmed using a cage-side analyzer (AlphaTRAK, Zoetis, UK). Serum and urine protein electrophoresis documented monoclonal (immunoglobulin M) gammopathy with Bence-Jones proteinuria. Computed tomography imaging revealed a monostotic osteolytic bone-lesion, and bone marrow cytology and histopathology documented plasmacytosis with multiple myeloma oncogene 1 / interferon regulatory factor 4 positivity, consistent with multiple myeloma. Infectious disease testing initially indicated seropositivity for Leishmania, Borrelia, and Anaplasma spp.; however, Leishmania PCR (splenic and bone marrow aspirates), and paired serological titers for Borrelia and Anaplasma were negative. Consequently, initial serological results were considered to be false positive because of paraproteinemia-associated assay interference. Chemotherapy (prednisolone and melphalan combination therapy) was initiated, but the dog was euthanased 30 days later because of the development of pericardial effusion. This is a report of spurious serological (and other laboratory) results occurring secondary to monoclonal gammopathy in a dog.

Multiple myeloma involving the gastrointestinal tract in an English Springer Spaniel.

A 10-year-old, entire male, English Springer Spaniel was referred for evaluation of weight loss, polyuria, polydipsia and gastrointestinal tract signs including melena/haematochezia for the previous six months. Results of serum protein electrophoresis, urine analysis, computed tomography of the thorax/abdomen, bone marrow aspiration and core biopsy, and splenic and mesenteric lymph node cytology were consistent with multiple myeloma. Endoscopically obtained gastrointestinal tract biopsies identified marked plasma cell infiltration within the duodenum, ileum and colon; immunohistochemistry showed positive labelling to MUM1 and Lambda confirming clonal plasma cell involvement. The dog entered complete clinical remission seven weeks after starting a melphalan/prednisolone protocol. The dog was euthanised 475 days after starting treatment due to cervical pain and collapse. At the time of euthanasia, blood work was not supportive of a relapse of multiple myeloma. To the authors' knowledge, this is the first report of multiple myeloma involving the gastrointestinal tract in a dog.

Comparative Oncology Assessment of a Novel Inhibitor of Valosin-Containing Protein in Tumor-Bearing Dogs.

Pet dogs with naturally occurring cancers play an important role in studies of cancer biology and drug development. We assessed tolerability, efficacy, and pharmacokinetic/pharmacodynamic relationships with a first-in-class small molecule inhibitor of valosin-containing protein (VCP/p97), CB-5339, administered to 24 tumor-bearing pet dogs. Tumor types assessed included solid malignancies, lymphomas, and multiple myeloma. Through a stepwise dose and schedule escalation schema, we determined the maximum tolerated dose to be 7.5 mg/kg when administered orally on a 4 days on, 3 days off schedule per week for 3 consecutive weeks. Adverse events were minimal and mainly related to the gastrointestinal system. Pharmacokinetic/pharmacodynamic data suggest a relationship between exposure and modulation of targets related to induction of the unfolded protein response, but not to tolerability of the agent. An efficacy signal was detected in 33% (2/6) of dogs with multiple myeloma, consistent with a mechanism of action relating to induction of proteotoxic stress in a tumor type with abundant protein production. Clinical trials of CB-5339 in humans with acute myelogenous leukemia and multiple myeloma are ongoing.

Erythrophagocytic multiple myeloma in a dog.

A 5-y-old spayed female Golden Retriever dog was referred because of severe normocytic normochromic nonregenerative anemia and thrombocytopenia. Serum analysis revealed hyperproteinemia and monoclonal or oligoclonal gammopathy. Fine-needle aspiration of the spleen revealed a highly erythrophagocytic population of neoplastic round cells, morphologically suggestive of plasma cells. After euthanasia, histologic assessment of the spleen and liver revealed an erythrophagocytic round cell tumor. Immunohistochemical analysis of the tumor population was positive for MUM1p and negative for CD3, CD20, and Iba-1, confirming the plasma cell origin of the tumor. Erythrophagocytic multiple myeloma is a very rare neoplastic condition in dogs.

Urinary bladder wall mass with neoplastic lymphoid cells in the urine: Diagnosis of an IgG secretory B-cell lymphoma with Bence-Jones proteinuria in a dog.

In this study, we describe a multimodal approach to diagnose a unique case of myeloma-related disease, extranodal secretory B-cell lymphoma with urinary bladder involvement, an IgG4 monoclonal gammopathy, and Bence-Jones proteinuria in a dog with a 6-year history of hyperglobulinemia that had not been further evaluated. A 12-year-old dog was presented for evaluation of a 1-week history of tenesmus. Urine sediment cytologic evaluation revealed low to moderate numbers of intermediate to large-sized lymphocytes. We describe a technique that yielded adequate numbers of viable neoplastic cells in shipped urine sediment for PARR and flow cytometry. Those studies demonstrated a clonal immunoglobulin gene rearrangement and an expansion of CD21-positive and MHC Class II-negative B cells, respectively. Protein electrophoresis with immunofixation and proteomic evaluation revealed a serum and urine IgG4 monoclonal gammopathy with Bence-Jones proteinuria. MUM1 immunocytochemistry performed on the urine sediment slides failed to label the neoplastic cells; thus, a plasma cell tumor was considered unlikely. Lack of response to a cyclophosphamide, vincristine, and prednisone chemotherapy regimen led to euthanasia without necropsy 21 days after diagnosis. Lymphoma is the most common hematopoietic malignancy and accounts for up to a quarter of all neoplasms in dogs, but lymphoid neoplasms arising primarily from extranodal sites are infrequently reported. Urinary tract neoplasia can be diagnosed by urine evaluation in about one-third of canine cases, but the diagnosis of lymphoid neoplasia via urine evaluation is rarely reported. This case highlights the utility of ancillary diagnostics on urine for detection of lymphoid malignancies.

Atypical multiple myeloma in 3 young dogs.

Three dogs under 12 months old were diagnosed with atypical multiple myeloma (MM), having an aggressive multifocal anaplastic round cell sarcoma in bone marrow, viscera, and/or peripheral blood, which were confirmed by cytology and immunohistochemistry to be of plasma cell origin. The intramedullary sarcomas caused myelophthisis, osteolysis, and hypercalcemia. Complete or free light chain monoclonal gammopathy in the serum and/or urine was demonstrated by protein electrophoresis and immunofixation. The polymerase chain reaction for antigen receptor rearrangement assay performed on 2 cases identified a clonally rearranged immunoglobulin gene. Neoplastic cells lacked expression of CD45, CD3, CD18, CD21, CD34, and MHCII by flow cytometry. Immunohistochemistry revealed MUM1 immunoreactivity of the neoplastic cells. Combining all data, the diagnosis was MM. An aggressive form of MM in young dogs should be a differential diagnosis for patients with an immunoglobulin-productive, B cell-clonal, CD45-negative, MUM1-positive discrete cell neoplasm arising from the bone marrow.

Solitary osseous plasmacytomas in dogs: 13 cases (2004-2019).

OBJECTIVES: To further characterise solitary osseous plasmacytoma in dogs, an extremely rare disease. To describe diagnosis, disease progression and treatment outcomes in dogs with solitary osseous plasmacytoma. MATERIALS AND METHODS: Retrospective review of dogs with solitary osseous plasmacytomas that were diagnosed and treated at a single institution from 2005 to 2019. Kaplan-Meier single group survival analysis was used to estimate median survival time and progression-free interval. RESULTS: Thirteen dogs met the inclusion criteria for the study, and of those, 11 were treated. The median age at diagnosis was 8 years (range 4 to 11). Most solitary osseous plasmacytomas occurred in the vertebrae (n=8). Other sites included the maxilla (n=2), the mandible (n=1), the tibia (n=1) and the carpus (n=1). The median survival time for all dogs with solitary osseous plasmacytoma was 912 days (range 5 to 2179), and the progression-free interval for treated dogs was 310 days (range 22 to 2179). Most dogs were treated with radiation therapy (n=10) with nine of 10 receiving a definitive, daily fractionated protocol and with five of ten having had neoadjuvant surgery. Seven dogs received chemotherapy, which was initiated after progressive disease in five dogs. The median survival time for dogs that completed radiation therapy (n=9) was 1166 days (range 545 to 2179). While five dogs developed lesions at other sites, no dogs progressed to multiple myeloma. CLINICAL SIGNIFICANCE: Canine solitary osseous plasmacytomas can be managed long term with appropriate local therapy. This observation reflects the biologic behaviour observed in humans.

Multiple myeloma and primary erythrocytosis in a dog.

A 13-year-old spayed female mixed breed dog was referred for impaired ambulation, limb tremors, back pain, hypergammaglobulinemia on cellulose acetate electrophoresis, and mild proteinuria. Conventional radiology and magnetic resonance imaging (MRI) suggested multifocal neoplastic bone lesions. At the referral examination, lameness and bright red mucous membranes were observed. Severe erythrocytosis, a monoclonal peak in the ß-2 globulin detected by capillary zone electrophoresis, severe proteinuria, bone marrow infiltration of plasma cells, and low serum erythropoietin concentrations were reported. The final diagnosis was multiple myeloma associated with severe primary erythrocytosis. This presentation in a dog is interesting because the combination of both disorders is rare in humans and has not been reported in dogs. Key clinical message: Although rare, multiple myeloma and primary erythrocytosis can occur together in dogs.

Myélome multiple et érythrocytose primaire chez un chien. Une chienne de race mixte stérilisée âgée de 13 ans a été référée pour troubles de la marche, tremblements des membres, maux de dos, hypergammaglobulinémie à l'électrophorèse sur acétate de cellulose et protéinurie légère. La radiologie conventionnelle et l'imagerie par résonance magnétique (IRM) suggéraient des lésions osseuses néoplasiques multifocales. Lors de l'examen de référence, une boiterie et des muqueuses rouge vif ont été observées. Une érythrocytose sévère, un pic monoclonal de la globuline ß-2 détecté par électrophorèse capillaire, une protéinurie sévère, une infiltration de la moëlle osseuse par des plasmocytes et de faibles concentrations sériques d'érythropoïétine ont été rapportés. Le diagnostic final était un myélome multiple associé à une érythrocytose primaire sévère. Cette présentation chez un chien est intéressante car l'association des deux conditions est rare chez l'homme et n'a pas été rapportée chez le chien.Message clinique clé :Bien que rares, le myélome multiple et l'érythrocytose primaire peuvent survenir simultanément chez le chien.(Traduit par Dr Serge Messier).

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.

BACKGROUND: The diagnostic performance of routine electrophoresis (agarose gel electrophoresis [AGE] and capillary zone electrophoresis [CZE]) and species-specific immunofixation (IF) for the detection of immunoglobulin paraproteins (M-proteins) and diagnosis of secretory myeloma-related disorders (sMRD) can be improved. Available canine IF targets were IgG-FC, IgA, IgM, light chain (LC), IgG4, and free LC (fLC) antibodies. OBJECTIVE: We aimed to review specific features associated with the presence of M-proteins in canine serum samples and the common features causing inaccurate reporting of M-proteins to improve the diagnostic performance of routine electrophoresis and IF for the detection of M-proteins. METHODS: Features found in AGE, CZE, routine IF, IgG4 IF, and fLC IF of 100 canine serum samples from Part 1 of this study were evaluated by simple and multivariate logistic regression to identify factors associated with the presence of M-proteins. Cases falsely called negative or positive for M-proteins were reviewed to identify the common features that could be used to increase the diagnostic performance of SPE and IF for M-protein detection. RESULTS: The presence of hypogammaglobulinemia or any peak taller than albumin was associated with an M-protein. Total protein concentrations, globulin concentrations, or peaks wider than albumin were not associated with an M-protein. Free LC sMRD cases were not diagnosed by SPE and routine IF. Cases with infectious and inflammatory etiologies had a restricted polyclonal gammopathy with multiple γ-globulin restrictions resulting in some false-positive results. SPE combined with all available IF results and the specific features identified in this study had an estimated sensitivity of 95.1% and specificity of 81.4%. CONCLUSIONS: The identified criteria of this study increase the diagnostic performance of the electrophoretic evaluation for M-proteins.

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.

BACKGROUND: Routine electrophoresis [agarose gel electrophoresis (AGE) and capillary zone electrophoresis (CZE)] and species-specific immunofixation (IF) can be used alone or in combination to detect immunoglobulin paraprotein (M-protein) and diagnose secretory myeloma-related disorders (sMRD). OBJECTIVE: We aimed to evaluate the performance of AGE, CZE, CZE plus IF (CZE-IF), and AGE plus IF (AGE-IF) for detecting canine serum M-proteins. METHODS: One hundred canine cases that had AGE, CZE, and routine IF performed on serum, and where B-cell lineage neoplasia (such as B-cell lymphoma and plasma cell tumors) had been diagnosed or excluded, were evaluated. Routine IF protocols targeted IgG-FC, IgA, and IgM heavy chains and light chains. IgG4 IF and free light chain IF were also performed. B-cell lineage neoplasms with an M-protein detected, using any available method, were classified as sMRD. Datasets from AGE, CZE, IF, CZE-IF, and AGE-IF (electrophoretograms, gel images, and fraction concentrations) were composed and reviewed. The sensitivity, specificity, and Youden's index for M-protein detection were determined for each dataset. RESULTS: The combination of AGE-IF or CZE-IF was more sensitive (82.9%) than CZE alone (72.0%) or AGE alone (64.6%) and more specific (66.1%, 48.3%, 51.7%, respectively). Immunofixation could be used alone to detect M-proteins (sensitivity 82.9%, specificity 61.9%), but there were technical challenges that complicated the performance and evaluation of the test. Myeloma with free light chains only was found in 5/41 cases of sMRD. CONCLUSIONS: Adding routine IF to routine electrophoresis increases the ability to accurately identify M-proteins; however, there is still room for further diagnostic performance improvements.

Light chain myeloma and detection of free light chains in serum and urine of dogs and cats.

BACKGROUND: Detection of free light chains (fLC) in animals relies on protein electrophoresis or the Bence-Jones protein test on urine. OBJECTIVE: To describe the detection of both serum fLC (sfLC) and urine fLC (ufLC) in 8 dogs and 2 cats using a commercially available human immunofixation (IF) kit. ANIMALS: Archived serum or urine samples from 27 dogs and 2 cats submitted to the Colorado State University Veterinary Diagnostic Laboratory for routine diagnostics. METHODS: Retrospective study evaluating the presence of fLC in dogs and cats using agarose gel electrophoresis and routine and fLC IF performed on serum and urine. The performance of the fLC IF reagents was evaluated using samples characterized by routine IF, tandem mass spectrometry, and a combination of fLC IF and western blotting. Free light chains were documented by paired electrophoresis and fLC IF. RESULTS: The fLC only myeloma case developed end-stage renal failure 5 months post initial diagnosis. All electrophoresis-defined urinary Bence-Jones proteins were labeled by the anti-free λ light chain (anti-fλ) reagent; none were labeled by the anti-free κ light chain (anti-fκ); 2 of these were identified as fκ by mass spectrometry. An electrophoretically identical protein restriction that was labeled by the anti-fλ reagent was present in the paired serum from 5/8 of cases, documenting sfLC. CONCLUSIONS AND CLINICAL IMPORTANCE: Commercially available human IF reagents identified sfLC and ufLC in both dogs and cats. Free light chains may be nephrotoxic in dogs.

Retrospective evaluation of the use of the International Myeloma Working Group response criteria in dogs with secretory multiple myeloma.

BACKGROUND: Current recommendations for monitoring disease progression and response to treatment in humans with multiple myeloma include evaluation of serum paraprotein (M-protein) concentration. Densitometry, species-specific radial immunodiffusion (RID) and ELISA methods can be used to quantify M-proteins. OBJECTIVE: Retrospectively evaluate use of the International Myeloma Working Group (IMWG) response criteria for humans in dogs with multiple myeloma. ANIMALS: Sixteen dogs with a diagnosis of multiple myeloma, M-protein documented by serum protein electrophoresis (SPE) and immunofixation (IF) in an initial sample and subsequent electrophoretic evaluation of serial samples. METHODS: Retrospectively, densitometric M-proteins, RID and globulins were measured and characterized according to IMWG criteria. Available clinical history was reviewed. Overall survival time (OST) was calculated from initial electrophoretic evaluation to death or last contact. RESULTS: All cases received some form of nonstandardized chemotherapy. Complete response (CR), a lack of detectable M-protein by SPE and IF, was documented in 1 case. Median survival was longer for dogs that attained ≥90% densitometric M-protein reduction (630 days) than for those that did not attain at least 50% reduction in densitometric M-protein (284 days; log rank P = .006). Five dogs were defined as having progressive disease (M-protein increase of >25% and at least 0.5 g/dL from nadir), which correlated with concurrent or subsequent clinical deterioration. Response criteria categorized by serum globulins or RID was not correlated with OST or clinical findings. CONCLUSIONS AND CLINICAL IMPORTANCE: Densitometric M-protein characterized using IMWG response criteria correlated with OST and clinical findings. Densitometric M-protein detection should be used to monitor dogs with multiple myeloma.