{"title":"利用流式细胞术中的 LT21 克隆 CD21 抗体诊断 CD21 表型阴性的犬 B 细胞慢性淋巴白血病:病例报告。","authors":"Eun Wha Choi, Yunho Jeong, Jin-Ok Ahn","doi":"10.1186/s12917-024-04335-x","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Chronic lymphoid leukemia (CLL) is a hematological disorder characterized by the clonal expansion of small mature lymphocytes that accumulate in the blood and bone marrow. CLL can arise from B-, T-, or natural killer cell clones. The cytological evaluation of blood smears is often the simplest and least invasive method for diagnosing lymphoid leukemia. Immunophenotyping is used to further subclassify the type of lymphoid leukemia.</p><p><strong>Case presentation: </strong>A 15-year-old, 4.4-kg spayed female Shih Tzu was presented to the veterinary medical teaching hospital of Kangwon National University. Despite having a normal appetite and activity level, cervical and inguinal lymph node enlargement was noted on physical examination. Complete blood count revealed severe leukocytosis, severe lymphocytosis, and monocytosis. Splenomegaly, hepatomegaly, and lymph node enlargement were detected on radiographic and ultrasonographic examination. Immunophenotyping was performed using peripheral blood mononuclear cells (PBMCs). The majority of lymphocytes exhibited the following profiles: CD3<sup>-</sup>CD79a<sup>-</sup> (97.5%), CD4<sup>-</sup>CD8<sup>-</sup> (98.6%), CD21<sup>-</sup>CD79a<sup>-</sup> (98.4%), CD34<sup>-</sup> (0.1%), CD45<sup>+</sup> (99.6%), major histocompatibility complex class II<sup>+</sup> (99.5%), and CD14<sup>-</sup> (0.5%). Based on the immunophenotyping results, possible differentials considered included the following: the majority of lymphocytes may be natural killer (NK) cell clones, plasma cell clones, or show aberrant expression or loss of CD21 marker due to the neoplastic nature of the cells. Further flow cytometry was performed using antibodies against CD3, CD5, CD94, and granzyme B. The combined results indicated that the predominant lymphocyte subset in the PBMCs was CD3<sup>-</sup>CD5<sup>-</sup>CD21<sup>-</sup>CD94<sup>-</sup>granzyme B<sup>-</sup>. To confirm monoclonality and exclude the aberrant loss of CD markers, a polymerase chain reaction for antigen receptor rearrangement (PARR) assay was conducted. The PARR assay, using DNA from blood and lymph node samples, showed B-cell monoclonality. Immunocytochemistry using PBMCs showed that the plasma cell marker Multiple Myeloma Oncogene 1 (MUM1) was not expressed. Therefore, the diagnosis was confirmed to be B-cell CLL.</p><p><strong>Conclusion: </strong>Immunophenotyping can help subclassify the type of lymphoid leukemia; however, as tumor cells can show aberrant expression or loss of the CD21 marker, combining immunophenotyping with the PARR assay could yield a more accurate diagnosis.</p>","PeriodicalId":9041,"journal":{"name":"BMC Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.3000,"publicationDate":"2024-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11515120/pdf/","citationCount":"0","resultStr":"{\"title\":\"Diagnosis of canine B-cell chronic lymphoid leukemia with a CD21 negative phenotype using the LT21 clone CD21 antibody in flow cytometry: a case report.\",\"authors\":\"Eun Wha Choi, Yunho Jeong, Jin-Ok Ahn\",\"doi\":\"10.1186/s12917-024-04335-x\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Chronic lymphoid leukemia (CLL) is a hematological disorder characterized by the clonal expansion of small mature lymphocytes that accumulate in the blood and bone marrow. CLL can arise from B-, T-, or natural killer cell clones. The cytological evaluation of blood smears is often the simplest and least invasive method for diagnosing lymphoid leukemia. Immunophenotyping is used to further subclassify the type of lymphoid leukemia.</p><p><strong>Case presentation: </strong>A 15-year-old, 4.4-kg spayed female Shih Tzu was presented to the veterinary medical teaching hospital of Kangwon National University. Despite having a normal appetite and activity level, cervical and inguinal lymph node enlargement was noted on physical examination. Complete blood count revealed severe leukocytosis, severe lymphocytosis, and monocytosis. Splenomegaly, hepatomegaly, and lymph node enlargement were detected on radiographic and ultrasonographic examination. Immunophenotyping was performed using peripheral blood mononuclear cells (PBMCs). The majority of lymphocytes exhibited the following profiles: CD3<sup>-</sup>CD79a<sup>-</sup> (97.5%), CD4<sup>-</sup>CD8<sup>-</sup> (98.6%), CD21<sup>-</sup>CD79a<sup>-</sup> (98.4%), CD34<sup>-</sup> (0.1%), CD45<sup>+</sup> (99.6%), major histocompatibility complex class II<sup>+</sup> (99.5%), and CD14<sup>-</sup> (0.5%). Based on the immunophenotyping results, possible differentials considered included the following: the majority of lymphocytes may be natural killer (NK) cell clones, plasma cell clones, or show aberrant expression or loss of CD21 marker due to the neoplastic nature of the cells. Further flow cytometry was performed using antibodies against CD3, CD5, CD94, and granzyme B. The combined results indicated that the predominant lymphocyte subset in the PBMCs was CD3<sup>-</sup>CD5<sup>-</sup>CD21<sup>-</sup>CD94<sup>-</sup>granzyme B<sup>-</sup>. To confirm monoclonality and exclude the aberrant loss of CD markers, a polymerase chain reaction for antigen receptor rearrangement (PARR) assay was conducted. The PARR assay, using DNA from blood and lymph node samples, showed B-cell monoclonality. Immunocytochemistry using PBMCs showed that the plasma cell marker Multiple Myeloma Oncogene 1 (MUM1) was not expressed. Therefore, the diagnosis was confirmed to be B-cell CLL.</p><p><strong>Conclusion: </strong>Immunophenotyping can help subclassify the type of lymphoid leukemia; however, as tumor cells can show aberrant expression or loss of the CD21 marker, combining immunophenotyping with the PARR assay could yield a more accurate diagnosis.</p>\",\"PeriodicalId\":9041,\"journal\":{\"name\":\"BMC Veterinary Research\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.3000,\"publicationDate\":\"2024-10-26\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11515120/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"BMC Veterinary Research\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1186/s12917-024-04335-x\",\"RegionNum\":2,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"VETERINARY SCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"BMC Veterinary Research","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1186/s12917-024-04335-x","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"VETERINARY SCIENCES","Score":null,"Total":0}
引用次数: 0
摘要
背景:慢性淋巴性白血病(CLL)是一种血液病,其特征是积聚在血液和骨髓中的成熟小淋巴细胞克隆性扩增。CLL 可由 B、T 或自然杀伤细胞克隆产生。对血液涂片进行细胞学评估通常是诊断淋巴性白血病最简单、侵入性最小的方法。免疫分型可用于进一步细分淋巴白血病的类型:江原国立大学兽医教学医院接诊了一只 15 岁、体重 4.4 千克的绝育雌性西施犬。尽管食欲和活动量正常,但体格检查时发现颈部和腹股沟淋巴结肿大。全血细胞计数显示白细胞、淋巴细胞和单核细胞严重增多。影像学和超声波检查发现脾肿大、肝肿大和淋巴结肿大。使用外周血单核细胞(PBMCs)进行了免疫分型。大多数淋巴细胞表现出以下特征:CD3-CD79a-(97.5%)、CD4-CD8-(98.6%)、CD21-CD79a-(98.4%)、CD34-(0.1%)、CD45+(99.6%)、主要组织相容性复合体 II 类+(99.5%)和 CD14-(0.5%)。根据免疫分型结果,可能的鉴别包括:大多数淋巴细胞可能是自然杀伤(NK)细胞克隆、浆细胞克隆,或由于细胞的肿瘤性质而出现 CD21 标记异常表达或丢失。使用 CD3、CD5、CD94 和颗粒酶 B 抗体进一步进行了流式细胞术检测。综合结果显示,PBMCs 中的主要淋巴细胞亚群为 CD3-CD5-CD21-CD94-颗粒酶 B-。为确认单克隆性并排除 CD 标记的异常丢失,进行了抗原受体重排聚合酶链反应(PARR)检测。使用血液和淋巴结样本的 DNA 进行的 PARR 检测显示 B 细胞为单克隆。使用 PBMCs 进行的免疫细胞化学分析显示,浆细胞标志物多发性骨髓瘤癌基因 1(MUM1)没有表达。因此,确诊为 B 细胞 CLL:结论:免疫分型有助于对淋巴性白血病的类型进行亚分类;但是,由于肿瘤细胞可能会出现 CD21 标记异常表达或缺失,因此将免疫分型与 PARR 检测相结合可得出更准确的诊断结果。
Diagnosis of canine B-cell chronic lymphoid leukemia with a CD21 negative phenotype using the LT21 clone CD21 antibody in flow cytometry: a case report.
Background: Chronic lymphoid leukemia (CLL) is a hematological disorder characterized by the clonal expansion of small mature lymphocytes that accumulate in the blood and bone marrow. CLL can arise from B-, T-, or natural killer cell clones. The cytological evaluation of blood smears is often the simplest and least invasive method for diagnosing lymphoid leukemia. Immunophenotyping is used to further subclassify the type of lymphoid leukemia.
Case presentation: A 15-year-old, 4.4-kg spayed female Shih Tzu was presented to the veterinary medical teaching hospital of Kangwon National University. Despite having a normal appetite and activity level, cervical and inguinal lymph node enlargement was noted on physical examination. Complete blood count revealed severe leukocytosis, severe lymphocytosis, and monocytosis. Splenomegaly, hepatomegaly, and lymph node enlargement were detected on radiographic and ultrasonographic examination. Immunophenotyping was performed using peripheral blood mononuclear cells (PBMCs). The majority of lymphocytes exhibited the following profiles: CD3-CD79a- (97.5%), CD4-CD8- (98.6%), CD21-CD79a- (98.4%), CD34- (0.1%), CD45+ (99.6%), major histocompatibility complex class II+ (99.5%), and CD14- (0.5%). Based on the immunophenotyping results, possible differentials considered included the following: the majority of lymphocytes may be natural killer (NK) cell clones, plasma cell clones, or show aberrant expression or loss of CD21 marker due to the neoplastic nature of the cells. Further flow cytometry was performed using antibodies against CD3, CD5, CD94, and granzyme B. The combined results indicated that the predominant lymphocyte subset in the PBMCs was CD3-CD5-CD21-CD94-granzyme B-. To confirm monoclonality and exclude the aberrant loss of CD markers, a polymerase chain reaction for antigen receptor rearrangement (PARR) assay was conducted. The PARR assay, using DNA from blood and lymph node samples, showed B-cell monoclonality. Immunocytochemistry using PBMCs showed that the plasma cell marker Multiple Myeloma Oncogene 1 (MUM1) was not expressed. Therefore, the diagnosis was confirmed to be B-cell CLL.
Conclusion: Immunophenotyping can help subclassify the type of lymphoid leukemia; however, as tumor cells can show aberrant expression or loss of the CD21 marker, combining immunophenotyping with the PARR assay could yield a more accurate diagnosis.
期刊介绍:
BMC Veterinary Research is an open access, peer-reviewed journal that considers articles on all aspects of veterinary science and medicine, including the epidemiology, diagnosis, prevention and treatment of medical conditions of domestic, companion, farm and wild animals, as well as the biomedical processes that underlie their health.