Converting fluid-based cytologic specimens to histologic specimens for immunohistochemistry.

BACKGROUND: Cavitary effusions are often evaluated cytologically to determine if there is an underlying neoplastic cause. Differentiation of neoplastic epithelial from mesothelial populations within effusions can be difficult using routine cytology. In addition, cytology alone cannot provide information on the immunophenotype of round cell populations. Gel foam techniques can be used to convert effusions into cell blocks for immunohistochemical (IHC) staining which can then be used to differentiate mesothelial from epithelial cell populations, and also allow immunophenotyping of round cell populations within effusions. OBJECTIVES: The aim of this study was to evaluate a gel foam cell block technique for converting potential neoplastic cells in cavitary effusions into cell blocks to characterize these further by IHC. RESULTS: Thirteen canine and 7 feline samples with cohesive cell populations were evaluated using gel foam cell blocks and IHC. Samples evaluated by routine cytology were categorized as (1) epithelial cells, (2) cohesive cell population without cytologic distinction between mesothelial and epithelial cells, and (3) mesothelial cells with signs of atypia. Antibody-mediated staining of vimentin and cytokeratin of the effusion cell blocks yielded further classification of cohesive cell populations. In addition, a total of 4 effusions with malignant round cells were evaluated; they were immunophenotyped as either B- or T-cell lymphoma. CONCLUSION: The use of cytokeratin and vimentin IHC on gel foam cell blocks from cavitary effusions provided robust staining and allowed characterization of cohesive cells as mesothelial or epithelial and immunophenotyping of lymphoid cell populations. In addition, this method is cost and time effective.

Significance of mastocytemia in cats.

BACKGROUND: In cats, mastocytemia is considered to be confined to animals with mast cell tumors (MCT), whereas in dogs it is associated with diverse diseases. OBJECTIVE: The objective of this retrospective study was to investigate the diagnostic and prognostic significance of mastocytemia in cats. METHODS: All blood smears and buffy coat (BC) smears on which mast cells were identified over a 6-year period were retrospectively reviewed and mast cells counted. Mastocytemic cats were classified based on their clinical diagnosis. RESULTS: Mastocytemia was identified on 40 blood smears and 13 BC smears from 33 cats. The incidence of mastocytemia detected in cats during routine CBCs was 0.33% (40/12,116 CBCs). Twenty-two of 33 mastocytemic cats (67%) had visceral (n = 17) or cutaneous MCT (n = 7), including 2 that had concurrent visceral and cutaneous involvement. In 3 additional cases (9%), visceral MCT was clinically suspected, but no cytologic or histopathologic evaluation of visceral organs was performed. MCT was excluded in 3 of 33 mastocytemic cats (9%) with a final diagnosis of lymphoid neoplasia (n = 2) and multiorgan hemangiosarcoma (n = 1). Five additional animals (15%) had a diagnosis other than MCT, including lymphoma (n = 2) and chronic renal failure (n = 3), but no cytologic or histopathologic evaluation of the spleen was performed. Blood smears from cats with confirmed MCT had 1-113 mast cells per smear, whereas cats in which MCT was excluded had 1-2 mast cells per smear. CONCLUSIONS: Data confirm that mastocytemia is rare and most commonly found in cats with visceral MCT; however, rare circulating mast cells may also be seen with neoplasms other than MCT.

Cytologic features of clear cell adnexal carcinoma in 3 dogs.

BACKGROUND: Clear cell adnexal carcinoma is a rare cutaneous adnexal neoplasm without definitive apocrine, sebaceous, or follicular differentiation. Differential diagnoses include sebaceous carcinoma, liposarcoma, and balloon cell melanoma. Immunohistochemical analysis, with positive immunoreactivity for pancytokeratin and vimentin, aids in the diagnosis. Cytologic features of clear cell adnexal carcinoma have not been described previously. OBJECTIVE: The aim of this study was to describe cytologic features of canine clear cell adnexal carcinoma. METHODS: Fine-needle aspirates (FNA) obtained prior to biopsy of cutaneous neoplasms with a histologic diagnosis of clear cell adnexal carcinoma confirmed by immunohistochemical analysis were reviewed retrospectively. Slides prepared from FNA had been stained with modified Wright-Giemsa or automated aqueous Romanowsky stain. RESULTS: Of 20 neoplasms diagnosed as clear cell adnexal carcinoma in dogs, FNA of the mass had been performed in 3 cases. Cytologic features were similar and included high cellularity, marked cellular pleomorphism, loose arrangement of neoplastic cells, and a light blue to gray background resulting from streaming of cytoplasm from ruptured cells. Neoplastic cells were oval to polygonal to spindle-shaped with wispy cytoplasmic projections. Cytoplasmic eosinophilic stippling, globular deposits, or pink needle-shaped inclusions were noted. Criteria of malignancy included marked anisocytosis, anisokaryosis and anisonucleoleosis, multinucleation, karyomegaly, macronucleoli, and atypical mitotic figures. CONCLUSIONS: Clear cell adnexal carcinoma should be included in the cytologic differential diagnosis for a canine cutaneous mass even if an epithelial origin is not readily identified owing to lack of characteristic epithelial features, such as highly cohesive cell clusters, evident cell-to-cell junctions, and distinct cytoplasmic edges.

The skin, a novel niche for recirculating B cells.

B cells infiltrate the skin in many chronic inflammatory diseases caused by autoimmunity or infection. Despite potential contribution to disease, skin-associated B cells remain poorly characterized. Using an ovine model of granulomatous skin inflammation, we demonstrate that B cells increase in the skin and skin-draining afferent lymph during inflammation. Surprisingly, skin B cells are a heterogeneous population that is distinct from lymph node B cells, with more large lymphocytes as well as B-1-like B cells that coexpress high levels of IgM and CD11b. Skin B cells have increased MHC class II, CD1, and CD80/86 expression compared with lymph node B cells, suggesting that they are well-suited for T cell activation at the site of inflammation. Furthermore, we show that skin accumulation of B cells and Ab-secreting cells during inflammation increases local Ab titers, which could augment host defense and autoimmunity. Although skin B cells express typical skin-homing receptors, such as E-selectin ligand and α-4 and ß-1 integrins, they are unresponsive to ligands for chemokine receptors associated with T cell homing into skin. Instead, skin B cells migrate toward the cutaneously expressed CCR6 ligand CCL20. Our data support a model in which B cells use CCR6-CCL20 to recirculate through the skin, fulfilling a novel role in skin immunity and inflammation.

Biodistribution and pharmacodynamics of recombinant human alpha-L-iduronidase (rhIDU) in mucopolysaccharidosis type I-affected cats following multiple intrathecal administrations.

The storage disorder mucopolysaccharidosis type I (MPS I) is caused by a deficiency in lysosomal α-L-iduronidase activity. The inability to degrade glycosaminoglycans (GAG) results in lysosomal accumulation and widespread tissue lesions. Many symptoms of MPS I are amenable to treatment with recombinant human α-L-iduronidase (rhIDU), however, peripherally administered rhIDU does not cross the blood-brain barrier and has no beneficial effects in the central nervous system (CNS). A feline model of MPS I was used to evaluate the CNS effects of rhIDU following repeated intrathecal (IT) administration. Twelve animals were randomized into four groups based on the time of euthanasia and tissue evaluation following three repeat IT administrations of 0.1 mg/kg rhIDU or placebo on Study Days 1, 4 or 5, and 9. Two days after the final IT injection, the mean tissue α-L-iduronidase (IDU) activity in the brains of the two treated animals were approximately 3-times higher (50.1 and 54.9 U/mg protein) than the activity found in normal cat brains (mean of 18.3 U/mg), and remained higher than untreated MPSI brain at 1 month (2.4 and 4.1 U/mg protein) before returning to near-baseline levels after 2 months. This activity corresponded with decreased brain GAG concentrations after 2 days (1.4 and 2.0 µg/mg) and 1 month (0.9 and 1.1 µg/mg) which approached levels observed in normal animals (0.7 µg/mg). Attenuation of GAG, gangliosides GM2 and GM3, and cholesterol reaccumulation was identified at both two days and one month following final IT injection. No adverse effects attributable to IT rhIDU administration were observed. IT rhIDU may be an effective means for providing enzyme replacement therapy for the central manifestations of MPS I.

Fibrosarcoma arising at the site of a retained surgical sponge in a cat.

An 8-year-old female spayed domestic shorthair cat had an abdominal mass palpated as an incidental finding on physical examination. Cytologic findings in ultrasound-guided fine-needle aspirates of the mass were most compatible with a sarcoma, with abundant mineralized material and mixed inflammation. The mass was removed surgically and on gross examination was white-tan, firm, associated with the mesentery, and when transected contained a gauze sponge in its center. On histopathologic examination, an area of central necrosis with mineralization and numerous refractile fibers consistent with sponge material was surrounded by dense fibrous connective tissue (gossypiboma). Within the connective tissue was a population of highly pleomorphic spindle cells consistent with a fibrosarcoma. Immunohistochemically, most neoplastic cells stained strongly positive for vimentin and a low number of cells were positive for smooth muscle actin. The results were consistent with a fibrosarcoma arising at the site of a retained surgical sponge. At a follow-up visit 2 months postoperatively, ultrasonographic and cytologic evidence of metastasis was found in the spleen and mesentery. To our knowledge, this is the first report of malignant transformation at the site of a retained surgical sponge in a cat and the first report of a fibrosarcoma arising within a gossypiboma in a domestic animal.

Multiple myeloma in 16 cats: a retrospective study.

BACKGROUND: There is limited published information regarding feline multiple myeloma. Diagnostic criteria are derived from canine studies and to our knowledge, have not been critically reviewed for cats. OBJECTIVE: To evaluate the clinical and laboratory findings in cats with multiple myeloma and appraise diagnostic criteria. METHODS: Retrospective evaluation of medical records was performed. Inclusion required an antemortem diagnosis of multiple myeloma using 2 of 4 criteria: 1) >or=20% plasma cells in the bone marrow, or >or=10% if atypical plasma cells; 2) paraproteinemia; 3) radiographically-evident osteolysis; 4) light chain proteinuria. Alternatively, a postmortem diagnosis was based on the findings of multiple plasma cell neoplasms, with marrow involvement. RESULTS: Sixteen cats were diagnosed with multiple myeloma between 1996 and 2004, with a median age of 14.0 years; 9 of 16 (56%) were castrated males, and 7 of 16 (44%) were spayed females. Laboratory abnormalities included hyperglobulinemia (14/16, 87.5%), with 11/14 (78.5%) monoclonal and 3/14 (21.4%) biclonal gammopathies; hypoalbuminemia (4/16, 25%); light chain proteinuria, (4/9, 44.4%); hypocholesterolemia (11/16, 68.7%); hypercalcemia, (3/15, 20%); nonregenerative anemia, (11/16, 68.7%); regenerative anemia, (1/16, 6.2%); neutropenia (5/15, 33.3%); thrombocytopenia (8/16, 50%); and marrow plasmacytosis (14/15, 93.3%). Plasma cells were markedly immature, atypical, or both in 10 of 12 (83.3%) cats. Focal or multifocal osteolysis was noted in 6 of 12 (50%) cats for which radiographs were available for review; generalized osteopenia was found in 1 (8.3%) cat. Noncutaneous, extramedullary tumors were found in all cats assessed, 7/7 (100%), including spleen (6), liver (3), and lymph nodes (4). The disease in 1 of 2 cats with cutaneous tumors progressed to plasmacytic leukemia. CONCLUSIONS: Common findings in feline multiple myeloma include atypical plasma cell morphology, hypocholesterolemia, anemia, bone lesions, and multi-organ involvement. Based on the results of this study, we advocate modifying diagnostic criteria in cats to include consideration of plasma cell morphology and visceral organ infiltration.