RESUMO
Reptile white blood cell (WBC) morphological features are strikingly variable across species. In the Argentine black and white tegu (Salvator merianae), red tegu (Salvator rufescens), and Savannah monitor (Varanus exanthematicus), previous reports described a WBC type with a single distinct, clear, linear- to ovoid- to crescent-shaped inclusion of presumptive monocytic origin. The objective of this study was to further investigate the origin of this unique WBC type with crescent-shaped inclusions. Blood samples from two Argentine black and white tegus, tegu 1, a 4-year-old female, and tegu 2, a 2-year-old presumed male, were submitted for routine hematological evaluation. Additional blood films were prepared and stained with these cytochemical stains: alkaline phosphatase (ALP; naphthol AS-MX phosphate substrate), alpha-naphthyl butyrate esterase, alpha-chloroacetate esterase, myeloperoxidase, Periodic acid-Schiff, and Sudan black B. Blood films from tegu 1 were also stained with a second ALP stain (5-bromo-4-chloro-3-indoxyl-phosphate and nitroblue tetrazolium substrate), Luna, luxol fast blue, and toluidine blue. The blood from tegu 1 was cytocentrifuged to isolate and fix the buffy coat in glutaraldehyde 2.5% aqueous solution for transmission electron microscopy. Six morphologically distinct WBC types were identified from tegu 1, including heterophils, basophils, monocytes, azurophils, lymphocytes, and the unique WBC type, which were identified as eosinophils with inclusions. WBC types in tegu 2 were similar; however, eosinophils lacked a discernable inclusion. Proper WBC identification will be useful in obtaining accurate hemogram data for this species.
RESUMO
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.
Assuntos
Linfócitos B , Doenças dos Cavalos/genética , Leucemia de Células B/veterinária , Doenças Linfáticas/veterinária , Linfopenia/veterinária , Transtornos Linfoproliferativos/veterinária , Paraproteinemias/veterinária , Animais , Antígenos CD19/análise , Linfócitos B/imunologia , Linfócitos B/metabolismo , Linfócitos B/patologia , Cavalos , Cadeias Pesadas de Imunoglobulinas/genética , Imunofenotipagem , Leucemia de Células B/genética , Leucemia de Células B/imunologia , Doenças Linfáticas/genética , Doenças Linfáticas/imunologia , Linfopenia/genética , Linfopenia/imunologia , Transtornos Linfoproliferativos/classificação , Transtornos Linfoproliferativos/genética , Transtornos Linfoproliferativos/imunologia , Fator de Transcrição PAX5/análise , Paraproteinemias/genética , Paraproteinemias/imunologia , Plasmócitos , Receptores de Complemento 3d/análise , Linfócitos TRESUMO
BACKGROUND: Detection of systemic inflammation, which is important for proper diagnosis and prompt treatment, can be challenging. HYPOTHESIS: Measurement of plasma iron concentration is a sensitive method for detecting systemic inflammation in horses compared with measurements of plasma fibrinogen concentration, a traditional marker for inflammation in the horse. ANIMALS: Ninety-seven horses hospitalized with diseases causing systemic inflammation, 22 horses with localized inflammation, and 12 clinically normal horses were included in this study. METHODS: A retrospective study was made on hospitalized horses that had both plasma iron and fibrinogen concentrations measured on hospital admission. RESULTS: Plasma iron concentration was lower in horses with systemic inflammation (64 +/- 45 microg/dL) than the reference interval minimum (105 microg/dL) and were significantly lower (P = .001) than the value in a group of horses with local inflammation (123 +/- 45 microg/dL) and in healthy transported horses (143 +/- 29 microg/dL). Low plasma iron and high fibrinogen concentrations were both sensitive indicators of systemic inflammation in horses with sensitivity of 90 and 82%, respectively. There was a similar correlation between either continued decreases in iron concentration (Rsp of 0.239) or increases in fibrinogen concentration (Rsp of 0.280) during hospitalization and a worse prognosis. CONCLUSIONS AND CLINICAL IMPORTANCE: Measurement of plasma iron concentration better reflected acute inflammation than did fibrinogen concentration.