Because the T-cell receptor gamma (TCRgamma) gene is rearranged at an early stage of T-cell development in both TCRalphabeta and TCRgammadelta lineages, it has been preferentially targeted to detect T-cell clonality in human lymphoma/leukemia. We isolated 22 independent cDNA clones encoding canine TCRgamma and the following analysis of nucleotide sequences using the dog genome database revealed that the canine TCRgamma locus contains at least four repertories of variable genes that can be organized into two distinct subgroups and six repertories of joining genes belonging to two distinct subgroups according to the nucleotide sequence similarity. The findings allowed us to design PCR primers that were directed to the conserved or specific nucleotide sequences for each subgroup of variable and joining genes. By using four different combinations of primers, a PCR-based analysis was performed on cell samples collected from T-cell lymphoma/leukemia and B-cell lymphoma cases and hyperplastic and normal lymph nodes. All cell samples from 11 T-cell malignancy cases exhibited clonal amplification by two out of four primer combinations. This finding was considered to be valuable in PCR-based analysis for detecting T-cell clonality in canine lymphoma/leukemia.
"The assessment of clonal rearrangements of antigen receptor genes has also been introduced to veterinary medicine (Vernau and Moore 1999; Burnett et al. 2003; Keller et al. 2004; Moore et al. 2005; Werner et al. 2005; Tamura et al. 2006; Yagihara et al. 2007; Thilakaratne et al. 2010). The methods described in some of these studies were reliably accurate which allow their use in clinical application. "
[Show abstract][Hide abstract] ABSTRACT: The objectives of this study were to detect clonal rearrangements of antigen receptor genes by polymerase chain reaction (PCR) assay and minimal residual disease (MRD) in clinical complete remission of chemotherapeutic-treated dogs. For PCR assays to determine clonality for antigen receptor rearrangement genes and MRD from cytologic and peripheral blood samples of 14 dogs with lymphoma either before chemotherapy and during remission, clonality was detected in 13 of the lymphomas before treatment. MRD was demonstrated in seven dogs with lymphoma during remission. Detection of MRD during remission in canine lymphoma using PCR technique is considered as a useful tool for prognosis and monitoring relapsing disease.
"According to a study, the rate of leukemic involvement of peripheral blood ranges from 28% to 65% in dogs with multicentric lymphoma, and depends on the stringency of criteria used (Keller et al, 2004). Thus, the assessment of clonal rearrangements of antigen receptor genes using PCR has been introduced into veterinary medicine, recently (Vernau and Moore, 1999; Burnett et al, 2003; Keller et al, 2004; Tamura et al, 2006; Yagihara et al, 2007). Also, in Korean veterinary clinics, diagnosis of canine B cell or T cell lymphoma using PCR was often carried out in case of uncertain if it is polyclonal or monoclonal lymphocytes. "
[Show abstract][Hide abstract] ABSTRACT: Lymphoma is the most common hematopoietic malignancy in dogs. Diagnosis of lymphoma is classically performed by morphological assessment and immunohistochemistry. But some cases in the early stage are difficult to distinguish and need more objective and accurate methods. So, Polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) and flow cytometric immunophenotype of lymphoma have been developed continuously. In this study, we performed these two methods to classify lymphoma type in 3 cases. According to PARR analysis, B cell origin lymphoma was diagnosed in two of three cases by testing PBMC and lymph node. All fine needle aspiration (FNA) samples of lymph nodes had high expression of CD21 on >88% of total cell population and PBMC samples also showed high expression of CD21 on >30% of total lymphocytes in those two cases, while the expression of CD3, CD4 and CD8 was absent. These results suggest that concurrent use of PARR and flow cytometric immunophenotype is more effective and valuable tool for the diagnosis and monitoring of canine lymphoma patients.
[Show abstract][Hide abstract] ABSTRACT: A novel canine lymphoma cell line, OSW, was established from the malignant pleural effusion of a dog with peripheral T-cell lymphoma. The immunoprofile as determined by flow cytometry was as follows: positive for CD45, CD49d, CD18, CD11a; weakly positive for CD11b, CD11c, CD11d; and negative for CD45RA, CD1a, CD1c, CD3, TCRalphabeta, TCRgammadelta, CD4, CD5, CD8a, CD8b, CD90(Thy1), CD21, MHCII, CD14(TUK4), CD34, and MPO. Immunocytochemistry of cytospin preparations was negative for cytoplasmic CD3, CD79a, and MPO, but was positive for CD20. The cell line had an oligoclonal T-cell receptor gamma (TCRgamma) gene rearrangement. Array comparative genomic hybridization (aCGH) and single locus probe (SLP) analysis showed that there were copy number increases of loci on dog chromosome 13 (CFA 13), and copy number decreases were evident for regions of CFA 11, 22, 26, 30 and 32, which include several of the more common chromosomal aberrations reported previously in canine lymphoma. The OSW cell line grows rapidly in vitro and is tumorigenic as a xenograft in SCID/NOD mice. OSW represents one of only a few reported canine lymphoma cell lines and is the most thoroughly characterized. This cell line and xenograft represent significant in vitro and in vivo models, respectively, for comparative and translational lymphoma research.
Leukemia Research 01/2008; 31(12):1709-20. DOI:10.1016/j.leukres.2007.04.003 · 2.35 Impact Factor
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