Classification of myeloid neoplasms: a comparative review

Classification of myeloid neoplasms in veterinary medicine was modeled in the early 1990s after French-American-British and National Cancer Institute systems used in human medicine. Recently our physician counterparts, in collaboration with oncologists, constructed a new World Health Organization (WHO) standard. WHO revisions lower the blast threshold from 30% to 20% for diagnosing acute myeloid leukemia (AML) and expand and redefine AML categories. AML is now subdivided into 4 broad groups: 1) AML with recurrent genetic abnormalities, 2) AML with multilineage dysplasia, 3) AML with previous chemotherapy and/or radiation, and 4) AML, not otherwise categorized. AML alphanumeric designations (M1, M2, etc) have been discontinued as numbers of subtypes have increased. The lower blast percentage eliminates one category of myelodysplastic syndrome (MDS): refractory anemia with excess blasts in transformation. A new MDS category was created: refractory cytopenia with multilineage dysplasia (RCMD), with lineage dysplasia assessed using newly defined percentage limits. At least 10% of cells from each of 2 lineages must display atypia for a diagnosis of RCMD. That threshold is 50% for diagnosing AML with multilineage dysplasia. Chronic myelomonocytic leukemia has been removed from the MDS category and included in a new category of diseases that have features of both MDS and chronic leukemia. WHO revisions are a signal to veterinary clinical pathologists to assess the validity of our system, which was built on premises now questioned.     

Subleukaemic acute myeloid leukaemia with myelodysplasia in a horse

Acute myeloid leukaemia (AML) is rarely reported in horses and myelodysplastic syndrome (MDS) has been described only in one case. Acute myeloid leukaemia is defined as the presence of at least 20% blasts in the marrow or blood. On the other hand, MDS is characterised by morphologic abnormalities in one or more cell lineages with hypercellular marrow and peripheral cytopenias due to ineffective haematopoiesis. We report a case of acute myelomonocytic leukaemia with myelodysplasia-related features in a horse. A supposed diagnosis was based on abnormal morphology of circulating neoplastic cells and bone marrow cytology. A final diagnosis was made by using flow cytometry (FC) in conjunction with cytochemistry (CC) rarely reported in the haematopoietic neoplasms of the horse.     

Myelodysplasia, hypophosphataemia, vitamin D and iron deficiency in an alpaca.

A pregnant 2-year-old alpaca was presented for evaluation of progressive weight loss, decreased appetite and lethargy that developed in winter. Haematologic and serum biochemical analyses revealed marked anaemia, leukopenia, severe hypophosphataemia and mild hypocalcaemia. Evaluation of bone marrow core biopsies and aspirates revealed an increased proportion of immature haematopoietic cells, without sufficient numbers of blast cells to be termed an acute myeloid leukaemia (AML). 1 The haematological and bone marrow findings were suggestive of myelodysplastic syndrome (MDS). The anaemia, leukopenia, lethargy and weight loss remained refractory to medical therapy and the alpaca was euthanased on humane grounds.     

Anti-CD71 antibody immunohistochemistry in the diagnosis of acute myeloid leukemia,subtype acute erythroid leukemia with erythroid dominance (AML M6-Er), in a retrovirus-negative cat

CD71 is an immunohistochemical marker used in diagnosing acute myeloid leukemia (AML) M6-Er in humans; however, to our knowledge, it has not been reportedly used for immunohistochemistry in veterinary medicine. We evaluated the pathologic features of AML M6-Er in a retrovirus-negative cat and used CD71 to support the diagnosis. A 4-y-old spayed female Scottish Fold cat was presented with lethargy, anorexia, and fever. Whole-blood PCR assay results for pro feline leukemia virus/pro feline immunodeficiency virus and feline vector-borne diseases were negative. Early erythroid precursors were observed in the peripheral blood smear. Fine-needle aspiration of the enlarged spleen and splenic lymph node showed many early erythroid precursors. Bone marrow aspirate smears revealed erythroid hyperplasia with 68.4% erythroid lineage and 3.6% rubriblasts. Dysplastic cells infiltrated other organs. The patient was diagnosed with myelodysplastic syndrome, progressing to the early phase of AML M6-Er. The patient died on day 121 despite multidrug treatments. Postmortem examination revealed neoplastic erythroblasts infiltrating the bone marrow and other organs. Neoplastic cells were immunopositive for CD71 but immunonegative for CD3, CD20, granzyme B, von Willebrand factor, CD61, myeloperoxidase, and Iba-1. Although further studies are necessary for the application of CD71, our results supported the morphologic diagnosis of AML M6-Er.     

Acute myeloblastic leukemia with associated BCR-ABL translocation in a dog

An 8-year-old male neutered Labrador Retriever was referred to the University of Wisconsin Veterinary Medical Teaching Hospital with a presumptive diagnosis of leukemia. Hematologic abnormalities included normal neutrophil count with a left shift, monocytosis, eosinophilia, thrombocytopenia, and circulating immature mononuclear cells. Bone marrow was effaced by immature hematopoietic cells of various morphologic appearances. In addition, large multinucleated cells were observed frequently. Flow cytometric analysis of nucleated cells in blood revealed 34% CD34(+) cells, consistent with acute leukemia. By immunocytochemical analysis of cells in blood and bone marrow, some mononuclear cells expressed CD18, myeloperoxidase, and CD11b, indicating myeloid origin; some, but not all, large multinucleated cells expressed CD117 and CD42b, the latter supporting megakaryocytic lineage. The diagnosis was acute myeloblastic leukemia without maturation (AML-M1). To identify genetic aberrations associated with this malignancy, cells from formalin-fixed paraffin-embedded bone marrow were analyzed cytogenetically by multicolor fluorescence in situ hybridization (FISH). Co-localization of bacterial artificial chromosome (BAC) containing BCR and ABL was evident in 32% of cells. This confirmed the presence of the canine BCR-ABL translocation or Raleigh chromosome. In people, the analogous translocation or Philadelphia chromosome is characteristic of chronic myelogenous leukemia (CML) and is rarely reported in AML. BCR-ABL translocation also has been identified in dogs with CML; however, to our knowledge this is the first report of AML with a BCR-ABL translocation in a domestic animal.     

Cabot rings as a result of severe dyserythropoiesis in a dog

An 11-year-old female Dachshund was presented with depression, diarrhea, weight loss, and radiographic evidence of masses involving the liver, spleen, and cranial lobe of the right lung. Results of a CBC included severe nonregenerative anemia (HCT 14.2%, hemoglobin, 4.3 g/dL, reticulocytes 66,000/microL) with marked metarubricytosis (nucleated RBCs 6.39 x 10(3)/microL). Examination of the peripheral blood smear revealed marked erythroid dysplasia, including marked anisocytosis with a prevalence of macrocytes, Howell-Jolly bodies, diffuse basophilic stippling, and multinucleated and atypical nucleated RBCs. Neutrophil hypersegmentation and giant forms were also noted. Numerous erythrocytes, particularly polychromatophilic cells, contained inclusions consistent with Cabot rings, which appeared as delicate red-purple ellipsoid or figure 8 structures. Rarely, Cabot rings were observed extracellularly. The dog was treated symptomatically with blood transfusions, prednisone, erythropoietin, and vitamin supplementation, but the anemia progressively worsened. The dog was euthanized 2 months after presentation. Bone marrow aspirate and core biopsy specimens obtained at the time of euthanasia revealed marked dysplastic changes in all cell lines, especially dyserythropoiesis, along with infiltrating carcinoma cells. A necropsy was performed, and histologic examination revealed poorly differentiated adenocarcinoma of the lung with multiple metastases to the marrow, spleen, and liver. The final diagnosis was marked myelodysplasia secondary to metastatic adenocarcinoma. Cabot rings are found rarely in humans with myelodysplasia, but have not been described previously in dogs. Based on the findings in this case, Cabot rings may occur rarely in dogs with severe dyserythropoiesis.     

MYELOID NEOPLASIA IN A DOG

A 4-year-old Scotch Collie bitch was presented for examination because of hyperthermia and anaemia. Haematological examination and bone marrow biopsy led to a diagnosis of myeloid neoplasia. At autopsy there was slight enlargement of the liver, spleen and some lymph nodes. Microscopic examination revealed extensive infiltration of these tissues by neoplastic myeloid cells.     

Tuberculosis in alpaca (Lama pacos) on a farm in Ireland. 1. A clinical report

This case report describes tuberculosis (TB) due to infection with Mycobacterium bovis (M. bovis) in alpaca (Lama pacos) on a farm in Ireland. Two severely debilitated alpaca were presented to the University Veterinary Hospital, University College Dublin in November 2004. Bloods were taken, and haematology and biochemistry results were indicative of chronic infection. Radiological examination showed evidence of diffuse granulomatous pneumonia suggestive of tuberculosis. On necropsy there were granulomatous lesions present throughout many body organs including lung, liver, kidney, intestine as well on peritoneum and mesentery. Culture of acid-fast bacilli from lesions led to a diagnosis of tuberculosis due to M. bovis. The use of intradermal skin testing proved inefficient and unreliable for ante mortem diagnosis of tuberculosis in alpaca. Infection due to M. bovis should be considered among the differential diagnoses of debilitating diseases in alpaca, particularly those farmed in areas known to be traditional black spots for tuberculosis in cattle.     

Bladder outlet obstruction in a 6-month-old alpaca secondary to pelvic displacement of the urinary bladder

A 6-month-old female alpaca was presented for stranguria. Based on the history, physical examination findings, and radiographic studies, the alpaca was diagnosed with bladder outlet obstruction, secondary to pelvic displacement of the bladder, a condition previously unreported in camelids. Cystopexy was performed and the alpaca recovered unremarkably.     

Renal dysplasia and urinary tract infection in a Bull Mastiff puppy

Renal dysplasia and concurrent pyelonephritis were diagnosed in a 15-week-old female Bull Mastiff which presented with clinical signs compatible with chronic renal failure. Ultrasound examination was consistent with renal dysplasia and clinical pathology supported a diagnosis of chronic renal failure and urinary tract infection. Renal histological examination demonstrated inflammation and scarring with concurrent tubular dysplasia and persisting primitive glomeruli, although the presence of chronic inflammation complicated interpretation. It is uncertain whether renal dysplasia was inherited or due to a urinary tract infection acquired in the neonatal period. Renal dysplasia has not previously been reported in the Bull Mastiff breed.     

Spontaneous hemangiosarcoma in the spleen and liver of a young rat

Spontaneous hemangiosarcoma in young rats is rare. In this report, we describe a case of a spontaneous hemangiosarcoma in the spleen and liver of young rats. At necropsy, multiple pale red masses were observed in the spleen. Histopathologically, solid growth and haphazardly arranged neoplastic cells were observed, although no characteristic growth pattern was observed. In contrast, irregularly sized small slit-shaped spaces containing erythrocytes were found among the neoplastic cells. Reticular fibers incompletely surrounding the neoplastic cells were observed by silver staining. Immunohistochemistry revealed that the neoplastic cells were positive for vWF and CD34. Electron microscopic examination revealed that the neoplastic cells had erythrocytes in the lumen and Weibel-Palade bodies in the cytoplasm and were arranged along a discontinuous basal lamina. These features indicate that the tumor originated from vascular endothelial cells. Based on these results, the tumor was diagnosed as a hemangiosarcoma in the spleen and liver.     

Bilateral cystic rete testis in an alpaca (Lama pacos).

A 9-year-old intact male alpaca (Lama pacos) was examined because of marked enlargement of the left scrotum. Ultrasound examination revealed a thin-walled anechoic structure in the area of the left testis. Aspirated fluid contained spermatozoa, many of which had abnormal morphology. Castration was performed and the left testis was markedly enlarged with a clear fluid-filled cyst. The cyst was lined by a single layer of squamous to cuboidal epithelial cells consistent with those originating from rete testis. The right testis was of a comparable size and shape to that of normal alpaca testis, but the rete testis was mildly to moderately dilated. Additional findings included chronic inflammation of the right testis and epididymis and epididymal fibrosis with ductal hyperplasia on the left. The diagnosis was bilateral cystic rete testis, most likely secondary to chronic inflammation.     

Small intestine large granular lymphoma in a horse

A 12-year-old Appaloosa gelding was referred to the Texas Veterinary Medical Center with a history of chronic diarrhea and weight loss. At necropsy, numerous oval, craterlike ulcers were observed throughout the small intestine. Histologically, these lesions were composed of a neoplastic proliferation of round cells with intracytoplasmic phosphotungstic acid-hematoxylin-positive granules. The tumor cells stained positively for the CD3 antigen and negatively for a B-cell marker. A diagnosis of large granular lymphoma was based on the morphologic and immunohistochemical characteristics of the neoplasm. The postmortem presentation of this case depicted unusual multifocal, ulcerative lymphomatous lesions throughout the small intestine without involvement of the regional lymph nodes. The histologic and ultrastructural morphology of the neoplastic lymphocytes was similar to that in previously reported cases of abdominal equine large granular lymphomas, but in this case the neoplasm was restricted to the small intestine.     

Proposed criteria for classification of acute myeloid leukemia in dogs and cats

Blood and bone marrow smears from 49 dogs and cats, believed to have myeloproliferative disorders (MPD), were examined by a panel of 10 clinical pathologists to develop proposals for classification of acute myeloid leukemia (AML) in these species. French-American-British (FAB) group and National Cancer Institute (NCI) workshop definitions and criteria developed for classification of AML in humans were adapted. Major modifications entailed revision of definitions of blast cells as applied to the dog and cat, broadening the scope of leukemia classification, and making provisions for differentiating erythremic myelosis and undifferentiated MPD. A consensus cytomorphologic diagnosis was reached in 39 (79.6%) cases comprising 26 of AML, 10 of myelodysplastic syndrome (MDS), and 3 of acute lymphoblastic leukemia (ALL). Diagnostic concordance for these diseases varied from 60 to 81% (mean 73.3 +/- 7.1%) and interobserver agreement ranged from 51.3 to 84.6% (mean 73.1 +/- 9.3%). Various subtypes of AML identified included Ml, M2, M4, M5a, M5b, and M6. Acute undifferentiated leukemia (AUL) was recognized as a specific entity. M3 was not encountered, but this subclass was retained as a diagnostic possibility. The designations M6Er and MDS-Er were introduced where the suffix "Er" indicated preponderance of erythroid component. Chief hematologic abnormalities included circulating blast cells in 98% of the cases, with 36.7% cases having >30% blast cells, and thrombocytopenia and anemia in approximately 86 to 88% of the cases. Bone marrow examination revealed panmyeloid dysplastic changes, particularly variable numbers of megaloblastoid rubriblasts and rubricytes in all AML subtypes and increased numbers of eosinophils in MDS. Cytochemical patterns of neutrophilic markers were evident in most cases of Ml and M2, while monocytic markers were primarily seen in M5a and M5b cases. It is proposed that well-prepared, Romanowsky-stained blood and bone marrow smears should be examined to determine blast cell types and percentages for cytomorphologic diagnosis of AML. Carefully selected areas of stained films presenting adequate cellular details should be used to count a minimum of 200 cells. In cases with borderline diagnosis, at least 500 cells should be counted. The identity of blast cells should be ascertained using appropriate cytochemical markers of neutrophilic, monocytic, and megakaryocytic differentiation. A blast cell count of > 30% in blood and/or bone marrow indicates AML or AUL, while a count of < 30% blasts in bone marrow suggests MDS, chronic myeloid leukemias, or even a leukemoid reaction. Myeloblasts, monoblasts, and megakaryoblasts comprise the blast cell count. The FAB approach with additional criteria should be used to distinguish AUL and various subtypes of AML (Ml to M7 and M6Er) and to differentiate MDS, MDS-ER, chronic myeloid leukemias, and leukemoid reaction. Bone marrow core biopsy and electron microscopy may be required to confirm the specific diagnosis. Immunophenotyping with lineage specific antibodies is in its infancy in veterinary medicine. Development of this technique is encouraged to establish an undisputed identity of blast cells. Validity of the proposed criteria needs to be substantiated in large prospective and retrospective studies. Similarly, clinical relevance of cytomorphologic, cytochemical, and immunophenotypic characterizations of AML in dogs and cats remains to be determined.     

Acute myeloid leukemia with mastocytic and megakaryocytic differentiation in a calf

A diagnosis of acute myeloid leukemia was made in a 10-month-old Holstein female calf. The leukemia was macroscopically characterized by great enlargement of the spleen and moderate enlargement of some lymph nodes. Histochemical and immunohistochemical examination disclosed the presence of neoplastic cells either containing metachromatic and tryptase-positive granules or expressing factor VIII-related antigen. The granules, which were positive for naphthol AS-D chloroacetate esterase and did not have particulate contents, were distinct from those of basophilic leukemia cells. This leukemia was thought to be derived from a common myeloid progenitor capable of giving rise to megakaryocyte-erythrocyte progenitors and granulocyte-monocyte progenitors with the ability to differentiate into mast cells.     

Calcific band keratopathy in an alpaca

A 4‐year‐old female Suri alpaca was presented for evaluation of acute onset weakness, lethargy, and recent development of opacities in both eyes. On ophthalmic examination, bilaterally symmetrical corneal opacities were noted along the interpalpebral fissures with a few corneal blood vessels intermingled. A presumed diagnosis of calcific band keratopathy was made based on location and appearance. The patient was euthanized a short while after diagnosis due to reasons unrelated to the eyes and histologic examination of the corneas revealed subepithelial calcium and vascularization, consistent with calcific band keratopathy. This case report is the first to document this ocular condition in an alpaca.     

A case of chronic granulocytic leukemia in a dog.

A case of chronic granulocytic leukemia was diagnosed in a ten year old miniature poodle and was observed for four and one half years. Methods of diagnosis and characteristic features are described. A persistent granulocytosis with a preponderance of mature forms and the absence of a detectable underlying pyogenic process were key diagnostic features which enabled distinction of this neoplastic process from acute granylocytic leukemia and a leukemoid reaction. Other features included dysplastic granulocytes in various developmental stages, marginal anemia and hyperplastic bone marrow (myeloid elements). No blast crisis occurred. This dog was euthanatized in August 1975.     

Identification of acute myeloid leukemia in dogs using flow cytometry with myeloperoxidase, MAC387, and a canine neutrophil-specific antibody

BACKGROUND: Flow cytometry may be used to determine immunophenotype or lineage of leukemic cells, but few antibodies are available that are specific for cells of monocytic and granulocytic lineage. OBJECTIVE: The purpose of this study was to evaluate the flow cytometric staining patterns of 3 commercial monoclonal antibodies for monocytes and granulocytes in clinically healthy dogs and in dogs with acute myeloid leukemia (AML). METHODS: Mouse antihuman macrophage antibody (MAC387), mouse anti-human myeloperoxidase (MPO), and a canine neutrophil-specific antibody (NSA) were evaluated using flow cytometry on blood from 6 clinically healthy control dogs, and on blood (n = 7) and/or bone marrow (n = 2) from 8 dogs with AML. A diagnosis of acute leukemia was confirmed by >30% blasts in bone marrow or >30% blasts in peripheral blood, together with bi- or pancytopenia, circulating CD34-positive blast cells, and clinical signs of disease. Leukemic samples also were evaluated using a wide panel of monoclonal antibodies. RESULTS: MAC387 stained neutrophils and monocytes from control dogs, although the staining profiles for the 2 cell types differed. MPO and NSA resulted in strong positive staining of neutrophils; MPO also stained monocytes weakly. Lymphocytes did not stain with any of the antibodies. One case was classified as AML of granulocytic lineage (AML-M1), 6 cases were classified as acute monocytic leukemia (AML-M5), and 1 case was classified as acute myelomonocytic leukemia (AML-M4). Neoplastic myeloblasts in the dog with granulocytic AML were positive for MPO, NSA, MAC387, and CD4. All monoblasts from the dogs with AML-M5 were positive for CD14, 5 of 6 were positive for MAC387, and 2 were positive for MPO. NSA staining was negative in the 2 dogs with AML-M5 in which it was evaluated. In the dog with AML-M4 variable percentages of blast cells were positive for CD14, MPO, MAC387, CD4, and NSA. CONCLUSIONS: Antigens identified by antibodies to MAC387, MPO, and NSA were expressed not just by normal mature neutrophils and monocytes, but also by neoplastic myeloblasts and monoblasts. These 3 antibodies may be useful as part of a wider panel for immunophenotyping AML in dogs.     

Malignant glucagonoma associated with hyperglycaemia,hypertriglyceridaemia, Candidiasis and bacterial pneumonia in a 12-year-old Arabian gelding

A 12-year-old Arabian gelding was presented for severe, rapid weight loss and polyuria. Physical examination revealed a body condition score of 1/9 with severe muscle wasting, moderate tachycardia, profound hyperglycaemia and hypertriglyceridaemia. The horse's clinical condition deteriorated rapidly resulting in euthanasia 16 h following admission. Post-mortem examination revealed multifocal nests of neoplastic endocrine cells within the liver which had immuno-positive glucagon reactivity. Based on the combination of cachexia, marked hyperglycaemia and glucagon-positive neoplastic cells, a diagnosis of malignant glucagonoma, which was presumed to be functional, was made. This is the first case of malignant glucagonoma to be reported in horses.     

Hydroxyurea-induced onychomadesis in a dog with chronic myeloid leukemia: A case report

A 12-year-old Rottweiler dog was presented with a history of prostration, weight loss and hyporexia for six months. Based on complete blood tests (hematological and biochemical analyses), bone marrow examination and imaging analysis, a diagnosis of chronic myeloid leukemia was made. Treatment with hydroxyurea at a dosage of 18?mg/kg twice daily was not effective in controlling the high count of white blood cells. Furthermore, after 35 days of hydroxyurea treatment, the animal developed onycholysis, with sloughing of the claws of the left pelvic and left thoracic limbs and exposure of the distal phalanx. Interruption of the medication was implemented, with clinical healing of the ungual lesions observed three months after initiation of the drug. White blood cells returned to normal after using cyclophosphamide. Currently, the animal is in complete remission, having a disease-free interval of 575 days without chemotherapy. To the authors' knowledge, this is the first report of hydroxyurea-induced onycholysis within a short-term period in a dog diagnosed with chronic myeloid leukemia.