• 대한수의학회지
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• 제61권3호
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• pp.21.1-21.6
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• 2021

Canine T-zone lymphoma (TZL) is a mature T-cell lymphoma in dogs. The diagnosis and sub-classification are impossible without biopsy or immunophenotyping by flow cytometry. An 11-year-old, spayed, female Golden Retriever presented with lymph node enlargement. Clinical examination was consistent with canine multicentric lymphoma. However, immunophenotyping revealed positive for CD3, CD4, CD5, CD8, CD21, TCRαβ, and MHCII but negative for CD34, CD45, CD79a, and TCRγδ. Histopathology revealed lymphocytes expanding to the cortex-preserving architecture and thinning of the nodal capsule, and CD3 positive but PAX-5 negative. Owing to the indolent nature of TZL, careful monitoring approach without clinical intervention was utilized.

• 한국임상수의학회지
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• 제40권4호
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• pp.298-302
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• 2023

A 7-year-old neutered male, domestic shorthair cat presented anorexia and lethargy. The complete blood cell count revealed severe non-regenerative anemia, lymphocytic leukocytosis, neutropenia, and thrombocytopenia. On the peripheral blood smear examination, medium to large lymphoblastic cells with moderate amounts of basophilic cytoplasm were observed in up to 70% of peripheral leukocytes. Feline leukemia and immunodeficiency viruses were not detected using a commercial diagnostic kit. While splenomegaly and blunt margins of the caudoventral liver were observed in abdominal radiography, changes in the intra-abdominal lymph nodes were not remarkable. Ultimately, flow cytometric immunophenotyping from the peripheral blood revealed a negative for B-cell markers (CD21-/CD79a-) and T-cell markers (CD3-/CD4-/CD5-/CD8-). Based on the hematological examination and the immunophenotyping assay, the cat was diagnosed with non-B, non-T acute lymphoblastic leukemia. Here, we report a rare case of non-B, non-T acute lymphoblastic leukemia to raise awareness and provide information on clinical symptoms and laboratory test and immunophenotyping analysis results.

• 대한수의학회지
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• 제58권1호
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• pp.33-37
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• 2018

Various trials have been conducted to develop therapies for serious untreatable diseases. Among these, those using stem cells have shown great promise, and adipose-derived mesenchymal stem cells (ADMSCs) are easier to obtain than other types of stem cells. Prior to clinical trials, characterization of ADMSCs with monoclonal antibodies should be performed. However, it is difficult to use species-specific antibodies for veterinarians. This study was conducted to confirm the panel of human antibodies applicable for use in immunophenotypic characterization of canine adipose-derived stem cells and feline ADMSCs extracted from subcutaneous adipose tissue collected during ovariohysterectomy. For flow cytometric immunophenotyping, the third passages of canine ADMSC and feline ADMSC and human CD31, CD34, CD42, CD44, CD62 and CD133 antibodies were used. Of these, CD133 reacted with canine cells (3.74%) and feline cells (1.34%). CD133 is known as a marker related with more primitive stem cell phenotype than other CD series. Because this human CD133 was not a species-specific antibody, accurate percentages of immunoreactivity were not confirmed. Nevertheless, the results of this study confirmed human CD133 as a meaningful marker in canine and feline ADMSCs.

• Asian Pacific Journal of Cancer Prevention
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• 제17권6호
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• pp.2909-2916
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• 2016

Background: Canine malignant lymphoma is classified into B- or T-cell origin, as in the human case. Due to differences in prognosis, a suitable method needs to be developed for lineage identification. Aims: To determine the accuracy of immunophenotypic and molecular information between three methods: immunocytochemistry (ICC), immunohistochemistry (IHC) and heteroduplex polymerase chain reaction for antigen receptor rearrangements (hPARR) in spontaneous canine lymphomas. Materials and Methods: Peripheral blood, fine needle aspiration and tissue biopsies from enlarged peripheral lymph nodes prior to treatment of 28 multicentric lymphoma patients were collected. Cytopathology and histopathology were examined and classified using the updated Kiel and WHO classifications, respectively. Anti-Pax5 and anti-CD3 antibodies as B- and T-cell markers were applied for immunophenotyping by ICC and IHC. Neoplastic lymphocytes from lymph node and white blood cell pellets from peripheral blood were evaluated by hPARR. Results: In this study, low grade B-cell lymphoma accounted for 25% (7/28), high grade B-cell lymphoma for 64.3% (18/28) and high grade T-cell lymphoma for 10.7% (3/28). According to the WHO classification, 50% of all cases were classified as diffuse large B-cell lymphoma. In addition, ICC showed concordant results with IHC; all B-cell lymphomas showed Pax5+/CD3, and all T-cell lymphomas exhibited Pax5-/CD3+. In contrast to hPARR, 12 B-cell lymphomas featured the IgH gene; seven presented the $TCR{\gamma}$ gene; five cases showed both IgH and $TCR{\gamma}$ genes, and one case were indeterminate. Three T-cell lymphomas showed the $TCR{\gamma}$ gene. The percentage agreement between hPARR and ICC/IHC was 60%. Conclusions: Immunophenotyping should not rely on a single method. ICC or IHC with hPARR should be used concurrently for immunophenotypic diagnosis in canine lymphomas.

• 한국임상수의학회지
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• 제37권6호
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• pp.350-354
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• 2020

Feline chronic lymphocytic leukemia (CLL) is a rare disease. Its diagnosis is not simple because of the absence of clinical signs and the presence of mature lymphocytosis. An 11-year-old female spayed Russian Blue cat was referred to the veterinary medical teaching hospital for lethargy, diarrhea, weight loss, and inappetence. Marked lymphocytic leukocytosis and a significantly increased number of small-to-intermediate-sized lymphocytes in the peripheral blood were found on hematological examination. The results of the feline leukemia virus and immunodeficiency virus test were negative. Further, mild splenomegaly was detected. Bone marrow aspirate analysis revealed mature lymphocytosis and a clonally rearranged T cell receptor gene with the polymerase chain reaction (PCR) for antigen receptor rearrangement assay. Flow cytometric immunophenotyping showed a homogeneous population of CD5+/CD21-T-cells in the peripheral blood and bone marrow. According to the results of the aforementioned examinations, CLL was diagnosed. Treatment was not initiated at the time of diagnosis because the clinical signs were mild and did not affect the quality of life. This report describes the clinical findings and use of advanced diagnostic tools such as molecular clonality analysis and immunophenotyping for the diagnosis of feline CLL.

• 대한수의학회지
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• 제63권4호
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• pp.38.1-38.6
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• 2023

Canine T-zone lymphoma (TZL) is an indolent form of T-cell lymphoma. Conservative management is usually recommended; however, chemotherapy may be considered for symptomatic or progressive cases. Herein, we describe two dogs with generalized peripheral lymphadenopathy and peripheral blood lymphocytosis at presentation. One dog presented with gross lesions on the tongue. Flow cytometric immunophenotyping and cytological examinations demonstrated findings consistent with those of TZL. Chemotherapy with chlorambucil and prednisolone was administered, which resulted in improvement of the condition without any adverse events. Chemotherapy with chlorambucil may be considered as an appropriate choice for treating canine TZL.

• IMMUNE NETWORK
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• 제13권2호
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• pp.43-54
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• 2013

Over the past 40 years, flow cytometry has emerged as a leading, application-rich technology that supports high-resolution characterization of individual cells which function in complex cellular networks such as the immune system. This brief overview highlights advances in multiparameter flow cytometric technologies and reagent applications for characterization and functional analysis of cells modulating the immune network. These advances significantly support highthroughput and high-content analyses and enable an integrated understanding of the cellular and molecular interactions that underlie complex biological systems.

• IMMUNE NETWORK
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• 제14권6호
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• pp.289-295
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• 2014

Flow cytometric immunophenotyping of peripheral blood lymphocyte subsets is a powerful tool for evaluating cellular immunity and monitoring immune-mediated diseases. The numbers and proportions of blood lymphocyte subsets are influenced by factors such as gender, age, ethnicity, and lifestyle. This study aimed to establish reference ranges for peripheral blood lymphocyte subsets in a healthy Korean population. Blood samples from 294 healthy adults were collected. Lymphocyte subsets were analyzed using a single-platform method with a flow cytometer; white blood cells and lymphocytes were analyzed using an automated hematology analyzer. The mean value of the white blood cell count was $5,665cells/{\mu}l$, and the mean values of the subtype counts (percentages) were as follows: lymphocytes, $1,928cells/{\mu}l$ (35.08%); $CD3^+$ cells, $1,305cells/{\mu}l$ (67.53%); $CD3^+CD4^+$ cells, $787cells/{\mu}l$ (40.55%); $CD3^+CD8^+$ cells, $479cells/{\mu}l$ (25.23%); $CD3^-CD19^+$ cells, $203cells/{\mu}l$ (10.43%); and $CD3^-CD56^+$ cells, $300cells/{\mu}l$ (15.63%). Additionally, the $CD4^+/CD8^+$ ratio was 1.81. In this study, gender and age significantly influenced blood lymphocyte subsets. Our results demonstrate that, as with other populations, a healthy Korean population has its own, region-specific, lymphocyte subset reference ranges.

• 대한수의학회지
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• 제48권3호
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• pp.363-367
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• 2008

A 4-year-old female Cocker spaniel was presented with respiratory distress and abdominal distension. Pleural effusion, ascites, hepatosplenomegaly, and superficial lymphadenopathy were observed and multicentric lymphoma was diagnosed by cytological examination. Immunophenotyping of lymph node and bone marrow using polymerase chain reaction for antigen receptor rearrangement identified a stage V lymphoma originating from T-cell. Despite of systemic chemotherapy using L-asparagenase, vincristine, cyclophoaphamide and prednisolone, neurologic deficits came out and progressed. Cerebrospinal fluid analysis revealed neoplastic lymphocytic pleocytosis indicating central nervous system involvement of lymphoma. The postmortem diagnosis was confirmed based on the histology and imunohistochemistry.

• 대한임상검사과학회지
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• 제54권1호
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• pp.68-72
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• 2022

혈관면역모세포성 T세포 림프종(AITL)은 성숙 T follicular helper cells의 림프증식성 질환이다. 80세 여성의 골수에서 비정형 림프구가 관찰되었고 B세포에 대한 유세포검사에서 CD19+, CD20-, CD22- 및 lambda 경쇄 restriction과 같은 비정상 표현형이 관찰되었다. 처음에는 유세포검사에서 관찰된 비정상 B세포에 대해 B세포 림프종을 의심하였다. 그러나 환자는 골수를 침범한 AITL로 진단되었다. 유세포검사의 재분석에서 소수의 비정상 T세포 군집을 발견했다. 표면 lambda 경쇄 restriction은 비특이적 결합으로 생각되었다. EBV 양성 T세포 림프종 환자에서 형질세포와 유사한 면역표현형을 가진 B세포가 관찰될 수 있다. 이러한 비정상 B세포 혹은 형질세포의 증식이 기저에 존재하는 T세포 림프종을 가릴 수 있다. 정확한 진단을 위해서는 통합적인 접근이 필요할 것이다.