Practical aspects of immunocytochemistry in canine lymphomas

The aim of this study was to evaluate the utility of immunocytochemistry in a standard veterinary practice and to determine the immunophenotype of tumor cells in cases of multicentric lymphoma in dogs by immunocytochemical analysis of fine-needle biopsy specimens. The study was performed on cytological samples collected from 54 dogs, in which multicentric lymphoma was recognised based on clinical data, cytology or cytology and histology, and follow-up information. Diagnosis of lymphoma was established according to the updated Kiel classification. Immunocytochemical assays were conducted using commercially available antibodies to the pan T-lymphocyte marker CD3 and B cell antigen receptor complex CD79 alpha. Among all animals examined B cell lymphoma was recognized in 42/54 (77.8%) of cases, while in the remaining 12/54 (22.2%) of dogs T cell lymphoma was recognized. In 11 animals with lymphoma recognized cytologically, in which an entire lymph node was obtained for histology, the results of routine cytology and immunocytochemistry fully corresponded with findings revealed by histology and immunohistochemistry. Immunocytochemistry can be successfully conducted in smears stored at room temperature for 24 hours without changes of staining results. It can be stated that application of standard cytopathological assessment in connection with immunocytochemistry of lymph nodes samples collected from dogs with lymphoma is a method of choice for establishing final diagnosis, and avoids the need for reexamination or collection of tissue samples for histopathology and immunohistochemistry during surgical procedures in ambiguous cases.     

Aberrant phenotypes and quantitative antigen expression in different subtypes of canine lymphoma by flow cytometry

Flow cytometry may be a useful tool to analyze lymphoma samples that are obtained from fine needle aspirations (FNA). This study aimed to determine if flow cytometric analysis add more objective and standardized information on the cellularity and morphology of lymphoma cells to conventional cytology. The typical immunophenotype of different lymphoma subtypes was assessed and leukocyte marker expression was evaluated to determine which antigens were more frequently over- or under-expressed in these lymphoma subtypes. Fifty FNA lymph node samples were evaluated from canine lymphomas. Thirty-one samples were identified to be of B-cell origin, sixteen were identified to be of T/NK-cell origin and three cases were classified as acute lymphoblastic leukaemia with lymph nodes involvement. The most common B-cell lymphoma subtypes were centroblastic lymphomas, whereas three cases were atypical and classified as B-large cell pleomorphic lymphomas. Among the T/NK lymphomas, small clear cells, large and small pleomorphic mixed cells, large granular lymphocytic cells and small pleomorphic cells were identified. Aberrant phenotypes and/or antigen under/over regulation was identified in thirty out of forty-seven lymphoma cases (64%; 18/31 B-cell=58% and 12/16 T-cell=75%). In B-cell lymphomas the most frequent finding was the diminished expression of CD79a (45%). CD34 expression was also observed in four cases (13%). Among T-cell lymphomas the prevalent unusual phenotype was the under-expression or absence of CD45 (25%). These findings reveal flow cytometry may be useful in confirming the diagnosis of lymphoma, as the technique allows one to add useful information about morphology of the neoplastic cells and identify antigenic markers and aberrant phenotypes.     

Cytopathology of canine lymphomas (100 cases)

Malignant lymphoma is one of the most common malignant tumours occurring in dogs. Fine needle aspiration biopsy (FNAB) is an excellent, specific diagnostic procedure used to assess pathological processes in lymph nodes. The aim of the present study was to conduct a cytopathological analysis of lymphoma in dogs and to analyse some epidemic aspects of occurrence of lymphoma in 100 dogs using Giemsa stained slides. Samples were obtained by fine needle aspiration biopsy, fine needle non-aspiration biopsy, lymph node impression smears and by examination of body cavity effusions. The determination of the type and subtype of tumour was made on the basis of the updated Kiel classification adopted for dogs. Based on cytopathological analysis, the lymphoma was diagnosed in 100 dogs: 44 were female and 56 male. The animals' age ranged from 1.5 year to 15 years (median: 7.5 years), the animals were of different breeds (72% of all dogs belonged to 28 different breeds) and crossbreeds (28%). In 29% of dogs the regional or general lymphadenomegaly was the only clinical sign observed, in remaining cases (71%) at least one abnormality connected to lymphoma was found. Among all diagnosed lymphomas, high-grade lymphomas were more prevalent (86% of all cases) than low-grade lymphomas (14% of all cases). The possibility of boxers having a predisposition to T cell lymphoma development could be also suspected.     

Lineage differentiation of canine lymphoma/leukemias and aberrant expression of CD molecules

Multiparameter flow cytometry analysis and specific cluster differentiation (CD) molecules were used to determine the expression profiles of B- and T-cell antigens on lymph node preparations from 59 dogs with generalized or multisystemic lymphoma. Lymph node samples from 11 healthy dogs were labeled to validate the specificity of antibodies and to formulate guidelines for interpretation of the results obtained from lymphoma samples. In normal lymph nodes, T-lymphocytes expressing CD3, CD4, or CD8 beta represented 59+/-11%, 43+/-8%, or 16+/-5% of the total cells, whereas B-lymphocytes expressing either CD21 or surface IgM (IgM) represented 37+/-9% or 14+/-5%, respectively. Small lymphocytes could be distinguished from large lymphocytes by forward light scatter. Of the patient samples 29 different breeds were represented with Golden and Labrador retriever being the most common. The lymphoma samples segregated into three groups based on CD antigen expression. Thirty cases predominantly expressed one or more combinations of CD79a, IgM, and CD21 representing a B-cell lineage. Three B-cell cases also expressed the stem cell antigen, CD34. Sixteen cases expressed one or more combinations of CD3, CD4, and CD8 consistent with a T-cell lineage and CD3+CD4+CD8--phenotype was the most common. Thirteen cases showed a mixed expression profile for T- and B-cell antigens and in three cases CD14 was highly expressed. Clinical response was poorest for T-cell lymphomas. Leukemic states occurred in all three phenotypes; but mixed cell cases had the greatest proportion. Dual immunofluorescence staining confirmed co-expression of T-cell (CD3) and B-cell antigens (CD79a or CD21) on neoplastic lymphocytes of six mixed cell cases. In one mixed cell case, dual immunostaining identified lymphocyte populations that stained mutually exclusive for CD79a and CD3. Six mixed cell lymphomas tested by PCR showed clonality for rearranged antigen receptor. Four cases that were CD79a+CD3+ had TCRgamma chain gene rearrangements, whereas two cases that were CD3+CD8+CD21+ had Ig heavy chain rearrangement. One case expressing multiple CD molecules (CD3+CD8+CD21+CD14+) was PCR negative for both Ig and TCRgamma gene rearrangement and could not be classified into a B- or T-cell lineage. We show for the first time co-expression of B- and T-cell markers on lymphoma cells that had specific T- or B-cell gene rearrangements. These findings suggest that aberrant CD molecule expression is not an uncommon finding in canine lymphomas and is a useful diagnostic marker for malignancy.     

Evaluation of Pax5 expression and comparison with BLA.36 and CD79αcy in feline non‐Hodgkin lymphoma

Paired box gene 5 (Pax5) is a widely used B‐cell marker for human and canine non‐Hodgkin's lymphoma (nHL); however, in the literature there is only one case report using Pax5 in a cat B‐cell lymphoma. The purposes of this study were to investigate the expression and detection of B‐cell specific activator protein (BSAP) using a monoclonal anti‐Pax5 antibody in feline nHL (FnHL) tissue samples to evaluate its diagnostic relevance as a B‐cell marker. A total of 45 FnHL samples in 45 cats were evaluated. B‐cell lymphoma was the most common immunophenotype (51.1%) for all the samples and T‐cell the most common immunophenotype (64.3%) for the gastrointestinal (GI) form. Pax5 stained 82.6% of all B‐cell lymphomas and no expression was found in any of the T‐cell lymphomas. Anti‐Pax5 antibody staining in FnHL is similar to that reported in human and canine counterparts and may offer an excellent B‐cell marker in cats.     

Canine indolent nodular lymphoma

Sixty-six cases of indolent canine lymphoid proliferation were reviewed. Age ranged from 1.5 to 16 years (median 9.0 years). Dogs of 26 breeds, plus 13 of mixed breeding or unknown lineage, were represented. B-Cell lymphomas (CD79a+) predominated. Marginal zone lymphoma (MZL), the largest group, involved lymph node (33 cases) and spleen (13 cases), with both tissues involved in five of these cases. Follicular lymphoma (FL) involved lymph nodes (five cases), and mantle cell lymphoma (MCL) occurred as solitary splenic masses (three cases). Nodal CD3+ T-zone lymphomas (TZL) (10 cases), were included since they resembled late-stage MZL at the architectural level. Two cases of marginal zone hyperplasia (MZH) were included to aid in differentiation of early MZL. Clonality status was determined in 54 cases by analysis of immunoglobulin heavy chain (IGH) and T-cell antigen receptor gamma (TCRG) gene rearrangement. Clonal rearrangement of IGH was detected in 28 of 35 MZL cases (80%), four of four FL cases (100%) and three of three MCL cases (100%). Concurrent cross lineage rearrangement of TCRG was detected in six MZL and two FL cases. Clonal rearrangement of TCRG was documented in five of eight TZL cases (63%). Limited survival data obtained for 18 dogs indicated that the B-cell lymphomas (MZL, MCL, and FL) and the T-cell lymphoma (TZL) were associated with indolent behavior and long survival. Although to the authors' knowledge, the true incidence of canine indolent lymphomas is unknown, the tumors are not rare and may have been underrecognized. Recognition of their architectural features, routine application of immunophenotyping, and molecular clonality assessment should alleviate this.     

High-throughput immunophenotyping of 43 ferret lymphomas using tissue microarray technology

To validate the use of the tissue microarray (TMA) method for immunophenotyping of ferret lymphomas, a TMA was constructed containing duplicate 1-mm cores sampled from 112 paraffin-embedded lymphoma tissue specimens obtained from 43 ferret lymphoma cases. Immunohistochemical (IHC) expression of CD3, CD79alpha, and Ki-67 (MIB-1) was determined by TMA and whole mount (WM) staining of each individual case for result comparison. There was a high correlation between CD79alpha and CD3 results comparing ferret TMA and WM sections (kappa statistic 0.71-0.73 for single-core TMA and 0.79-0.95 for duplicate-core TMA) and between continuous data from Ki-67 staining of ferret TMA sections and WM sections (concordance correlation coefficients 0.77 for single cores and 0.87 for duplicate cores). Subsequently, a panel of commercially available antibodies was applied to the TMA for the analysis of expression in ferret lymphomas. The results of this study confirmed previously published results suggesting specific cross-reactivity of the applied IHC markers (CD3, CD79alpha, Ki67) with ferret lymphoma tissue. Other IHC markers (CD45Ro, bcl2, bcl10, MUM1, CD30, vimentin) were also expressed in subsets of the included ferret lymphomas. Further studies are necessary to determine the usefulness of these markers for diagnostic and prognostic evaluation of ferret lymphomas. In conclusion, the TMA technology was useful for rapid and accurate analysis of protein expression in large archival cohorts of ferret lymphoma cases.     

Histopathologic classification of 171 cases of canine and feline non-Hodgkin lymphoma according to the WHO

A retrospective collection of 171 lymphoid neoplasms (123 dogs and 48 cats) was classified according to the Revised European–American Lymphoma (REAL) classification, adopted in 2002 by the World Health Organization (WHO), to evaluate the WHO system for categorization of canine and feline neoplasms. Microscopic examination was performed after standard hematoxylin–eosin staining and immunohistochemical labelling for B (CD79a) or T (CD3) cell phenotypes. B-cell lymphomas were prevalent in dogs and T-cell lymphomas in cats. B-Large cell lymphoma (B-LCL) frequently showed plasmacytoid differentiation; notably, two canine plasma cell tumours (PCT) expressed both CD79 and CD3. There were difficulties in differentiating B-lymphoblastic lymphoma (B-LBL) from Burkitt-type lymphoma. Furthermore, intestinal T-cell lymphoma (ITCL) exhibited a huge morphologic variability. Finally, multicentric mature small and thymic T-cell lymphomas were diagnosed, although these categories are not codified by the WHO classification.     

Immunohistochemical detection of multiple myeloma 1/interferon regulatory factor 4 (MUM1/IRF-4) in canine plasmacytoma: comparison with CD79a and CD20

Multiple myeloma oncogene 1/interferon regulatory factor 4 (MUM1/IRF4) is involved in lymphoid cell differentiation, particularly in the production of plasma cells. We examined the immunoreactivity of mouse monoclonal antibody Mum-1p to MUM1/IRF4 and compared it with expression of CD79a and CD20 in 109 plasmacytomas in 107 dogs. Tissues had been fixed in formalin and embedded in paraffin. One hundred one of 109 (93.5%) tumors were positive for MUM1/IRF4. The staining was nuclear with weak cytoplasmic reaction. Fifty-nine of 105 (56.2%) plasmacytomas were positive for CD79a; only 21 of 108 (19.4%) cases were positive for CD20. MUM1/IRF4 staining was performed on 139 other tumors including B- and T-cell lymphomas, histiocytic proliferations, mast cell tumors, and melanocytic tumors. The only MUM1/IRF4-positive nonplasmacytic tumors were 10 B-cell lymphomas and 1 anaplastic lymphoma. We conclude the following: 1) Antibody Mum-1p is very specific for canine plasmacytomas, 2) antibody Mum-1p is superior in sensitivity and specificity to CD79a and CD20 for the identification of canine plasmacytomas in formalin-fixed, paraffin-embedded tissues, 3) canine lymphomas that express MUM1/IRF4 are few and usually of B-cell origin, 4) other canine leukocytic and melanocytic tumors do not express MUM1/IRF4, and 5) prospective studies are needed to determine whether the expression of MUM1/IRF4, particularly in lymphomas, has prognostic significance.     

Prognostic significance of morphological subtypes in canine malignant lymphomas during chemotherapy

The aim of this study was to determine the response of different morphological subtypes of canine lymphoma to a standardized therapeutic protocol. Diagnosis of lymphoma was based on cytohistological analysis and immunophenotyping with antibodies against CD3 and CD79a of an enlarged lymph node or an extranodal mass. Fifty-seven cases were classified according to the updated Kiel classification adapted to the canine species, into 24 B-cell lymphomas (20 centroblastic polymorphic and four Burkitt-type subtypes), and 33 T-cell lymphomas (10 pleomorphic mixed, 10 lymphoblastic, eight unclassifiable high grade plasmacytoid, and five small clear-cell subtypes). All dogs were clinically staged at diagnosis. The protocol used l-asparaginase, vincristine, cyclophosphamide, doxorubicin, and prednisone. First remission duration and overall survival time were evaluated. Although the T-cell phenotype was associated, on the whole, with a poor prognosis, as previously reported in veterinary and human medicine, the study showed significant prognostic differences between the B- and the T-cell subtypes of canine lymphoma and suggests that clinico-morphological characterization of the disease is justified in dogs, as in humans.     

Characterization of lymphocytes in canine gastrointestinal lymphoma

Primary canine gastrointestinal lymphoma has been believed to be of B-cell origin based on the morphology and behavior of the neoplastic cells and the evidence from the human medical field. However, the neoplasms have not to date been characterized as to the origin of the cell population. Forty-four cases diagnosed as canine gastrointestinal lymphoma were retrieved from the records of the Veterinary Teaching Hospitals at the University of Minnesota and the University of Wisconsin-Madison. Four of the cases have been previously identified as epitheliotropic T-cell gastrointestinal lymphoma. Twenty-three of the dogs were female, with 11 intact and 12 neutered, and 21 of the dogs were male, with 12 intact and nine neutered. Sixteen breeds as well as individuals of mixed breeding were represented. The Boxer and the sharpei were the most commonly represented breeds with six individuals each. The age range of the dogs was 1.5-14.66 years, with two dogs identified as adult and two of unknown age. Archived tissue blocks of gastrointestinal samples were sectioned in duplicate and prepared for immunohistochemical staining with CD3 (T-cell marker) and CD20 (B-cell marker). In 75% of the cases examined under light microscopy, 50-95% of the neoplastic cells stained positively with CD3 and exhibited marked epitheliotropic behavior. In three of the cases, from 10% up to 50% of the neoplastic cells stained positively with CD20, with widely scattered CD3(+) cells. In the remainder of the cases, few to none of the neoplastic cells stained with either of the markers. This retrospective study shows that canine primary gastrointestinal lymphoma is more commonly of T-cell origin, rather than B-cell origin.     

Immunohistochemical Detection of Retinoid Receptors in Tumors from 30 Dogs Diagnosed with Cutaneous Lymphoma

Background: Retinoids exert their effects by binding to retinoid receptors. Two types of retinoid receptors have been described: retinoic acid receptor (RAR) and retinoid X receptor (RXR), and their subtypes α, β, and γ. The expression of subtypes varies depending on the disease process. This study intended to detect the pattern of retinoid receptor expression in cutaneous lymphomas in dogs. Hypothesis: Cutaneous lymphomas in dogs have variable expression of retinoid and retinoid X receptors. Animals: Biopsy specimens from 30 dogs with cutaneous lymphoma. Methods: Tissues of dogs with cutaneous lymphoma were evaluated by immunohistochemistry for expression of retinoid receptors. The tissues were tested for the presence of 3 RAR and RXR subtypes (α, β, and γ). Lymphoma immunophenotype was determined by the use of the immunohistochemical markers CD79a (B‐cell) and CD3 (T‐cell) in all cases. Results: Twenty‐nine of 30 dogs were CD3 positive. The retinoid receptors expressed with the greatest frequency were RARβ (87% of cases), and RXRα and RXRγ (77% of cases). The expression of RARγ was not observed. Conclusions and Clinical Relevance: Retinoid and rexinoid receptor binding drugs may have an impact on the treatment of dogs with cutaneous lymphoma.     

Prognostic factors for treated canine malignant lymphoma.

The aim of this study was to investigate the prognostic importance of different clinical, immunohistologic and tumor proliferation characteristics in dogs with malignant lymphoma treated with chemotherapy. From 74 dogs with malignant lymphoma at least one enlarged peripheral lymph node was taken for biopsy before chemotherapy following a standardized protocol (vincristine, cyclophosphamide, prednisolone, doxorubicin, and L-asparaginase). The variables evaluated as prognostic factors were age, sex, and tumor stage, as well as histomorphologic grade (Kiel classification, Working Formulation), immunophenotype (using markers for CD3 and CD79a), and cell proliferation (Ki-67, proliferation cell nuclear antigen, mitotic index, and argyrophil nucleolar organizer regions [AgNORs]) in extirpated lymph nodes. All markers were used on routinely formalin-fixed, paraffin-embedded tissues. The AgNORs were assessed qualitatively, based on the AgNOR pattern distribution, and quantitatively using image analysis and routine counting. In both univariate and multivariate survival analyses, AgNORs were a valuable prognostic marker for the treatment of canine malignant lymphomas. Based on the results of the multivariate analysis longer survival time correlated with a B-cell type, a larger mean AgNOR area, a larger total AgNOR area, a shorter distance between two AgNORs, and a smaller AgNOR area to nucleus ratio. Longer disease-free survival time correlated with a smaller number of AgNORs per nucleus, a larger mean AgNOR area, a larger maximal AgNOR area, and a larger total AgNOR area. This study clearly demonstrates the additional benefit of the use of AgNORs in predicting treatment outcome in dogs with malignant lymphoma.     

Malignant lymphoma in nasal cavity and paranasal sinuses of a dog

A case of a canine large cell type T-cell lymphoma, with features of high-grade malignancy is described. The tumour was found confined in the nasal cavity and the paranasal sinuses of a crossbred German Shepherd dog. Histological examination revealed the features of a highly malignant large cell lymphoma. Ultrastructurally, the lymphoid tumour cells bore cytoplasmic protrusions that interdigitated tightly. From a panel of tumour markers used, the neoplastic cells were stained only for vimentin. Immunophenotyping of the tumour cells by means of CD3, CD79, kappa-light chains and lambda-light chains detection was undertaken. The tumour stained only for CD3 and was classified as T-cell lymphoma.     

Primary lymphoma of the uterine horn in a Lhasa Apso dog

Primary lymphomas of the canine female genital tract are uncommon tumours. A 9-year-old intact female Lhasa Apso dog presenting with a closed pyometra underwent an ovariohysterectomy (OHE), and the hyperplastic uterine horn along with multiple follicular cysts on the right ovary was examined by histological analysis. Severe infiltration of medium-sized lymphocytes with strong positive immunoreactivity for CD79a and numerous anaplastic features was detected in the unilateral uterine horn, and the dog was diagnosed as having extranodal marginal zone B-cell lymphoma (MZBCL). The present case reports an extremely rare occurrence of primary lymphoma involving the uterine horn in a dog and describes histological characteristics of the tumour for definite diagnosis.     

Lymphoid neoplasms in swine

Seventeen cases of lymphoid neoplasms in swine were investigated and divided into eight histological types. Cases 1-3 were precursor B lymphoblastic leukemias, which occurred in three piglets from the same dam. Cases 4 and 5 were diagnosed, respectively, as a precursor B lymphoblastic lymphoma and a thymic B cell lymphoma, because there were cytological differences between the lymphomas. These five cases of immature B cell malignancies expressed CD79a and terminal deoxynucleotidyl transferase (TdT). Mature B cell lymphomas were divisible into follicular (case 6), diffuse centroblastic (case 7) and intestinal large B cell (cases 8-11) lymphomas. Unlike in case 7, the neoplastic cells in cases 8-11 showed cytological features intermediate between centroblasts and immunoblasts. The mature lymphomas were characterized by positive immunolabeling for CD79a and cytoplasmic immunoglobulins. A case of thymic γδ T cell lymphoma (case 12) were positive for CD3, CD5, WC1 and TdT. Instead of TdT, perforin was expressed in γδ T cell lymphomas (cases 13-17), whose histological characteristics were epitheliotropism, homing into T cell zones of lymphatic tissues, and cytological atypia and pleomorphism. In the present study, lymphoid neoplasms could be classified into discrete histological types, some of which were considered to be specific for swine.     

The cloning and expression of matrix metalloproteinase-2 and tissue inhibitor of matrix metalloproteinase 2 in normal canine lymph nodes and in canine lymphoma

Matrix metalloproteinase-2 (MMP-2) and its inhibitor, tissue inhibitor of matrix metalloproteinase 2 (TIMP2), are known to be important in cancer. The purposes of this study were to determine the cDNA sequence of canine MMP-2 and to investigate the expression patterns of MMP-2 and TIMP2 in normal canine lymph nodes and spontaneously arising canine lymphomas. We cloned and sequenced a PCR product containing most (1901 base pairs) of the coding sequence of canine MMP-2 that translates into a 623 amino acid protein. The cDNA and deduced amino acid sequences are highly homologous to those of other mammalian species. Canine MMP-2 and TIMP2 mRNAs were detectable in the majority of normal lymph node and lymphomatous samples evaluated. No statistical difference was identified when comparing the expression of either gene with regard to normal versus neoplastic nodes, nodal versus extranodal lymphoma, lymphoma grade, or B versus T cell immunophenotype.     

The animal‐dependent risk factors in canine T‐cell lymphomas

Malignant lymphomas are one of the most common malignancies occurring in dogs; among them T‐cell tumours are less commonly recognized. Recently, many authors have recommended cytology as a sufficient diagnostic method for canine lymphomas, especially if supported by immunocytochemistry or flow cytometry. The aim of the study was to characterize animal‐dependent risk factors in canine T‐cell lymphomas (TCLs) in Poland, including specific cytological subtypes. Determination of the type and subtype of the tumour was made based on the updated Kiel cytological classification adopted for dogs as previously described. Two breeds turned out predisposed to TCL (dog de Bordeaux and Boxer) while no predisposition to B‐cell lymphoma could be evidenced. Dogs with low‐grade lymphoma were significantly older than those with high‐grade lymphoma.     

Canine T-cell lymphomas: a morphological,immunological, and clinical study of 46 new cases

The aim of this study is to report 46 new cases of canine T-cell lymphomas among a series of 140 lymphomas studied by immunophenotyping (incidence 32.8%). According to the updated Kiel classification adapted to the canine species, 13 were classified as low-grade and 33 as high-grade lymphomas. Among the low-grade lymphomas, five were small clear-cell lymphomas, three were pleomorphic small-cell lymphomas, and five mycosis fungoides. Among the high-grade cases, there were 11 pleomorphic mixed-, small-, and large-cell lymphomas, 6 pleomorphic large-cell lymphomas, 11 lymphoblastic lymphomas, and 5 unclassifiable high-grade plasmacytoid lymphomas. The cytohistologic features were highly suggestive of a T-cell phenotype on the basis of cell morphology (irregular nuclei and clear cytoplasms) (30/46 cases), a T-cell zone pattern, and the presence of hyperplastic postcapillary venules (22/46 cases). All 46 cases were CD3+ CD79a-, and among 34 cases investigated for CD4 and CD8 expression, 13 were CD4+CD8-, 13 were CD8+CD4-, and 8 were CD4CD8 double positive or double negative. The pleomorphic mixed lymphomas were mainly CD4+CD8- (6/7) and the lymphoblastic lymphomas were double positive or double negative (6/8). The main clinical, hematologic, and biochemical features were generalized (28/46) or regional lymphadenopathy (16/46), hepatosplenomegaly (15/46), extranodal involvement (11/46), mediastinal mass (9/46), and leukemia (8/46), which were mainly present in cases of lymphoblastic lymphomas and hypercalcemia (16/46).     

Immunophenotyping lymphomas in dogs: a comparison of results from fine needle aspirate and needle biopsy samples

The reliability of utilizing cytologic samples for immunophenotyping canine lymphomas was evaluated by a systematic comparison of results from fine needle aspirate (FNA) to needle biopsy specimens stained by immunochemical methods. The specific reactivity of a selected panel of 12 antibodies to cell surface markers and intermediate filaments was assessed in 11 dogs by comparing cytologic to histologic samples. There was excellent correlation of results between immunostained cytologic and histologic samples. FNA sampling is a simple, noninvasive method for determining the lymphoid phenotype in canine lymphoma. In addition, this technique may be useful as a cytodiagnostic aid in differentiating lymphoid from non-lymphoid tumors and in assessing neoplastic vs. reactive or hyperplastic processes.