Anti-Tcrg monoclonal antibody [APC]

A 9-year-old female spayed English Springer Spaniel was evaluated for a cranial mediastinal mass and lymphocytosis. Flow cytometric immunophenotyping of peripheral blood lymphocytes revealed 97% as CD3 positive, confirming a T-cell lineage. Additionally, T-cell subset assessment showed 53.2% to be double-negative T-lymphocytes, expressing neither CD4 nor CD8 surface markers. The number of double-negative lymphocytes in circulation coincided with the number of T-cell receptor (TCR) gamma delta-expressing T-cells in circulation. Molecular T-cell clonality analysis of TCR Gamma (TCRG) gene rearrangement showed a polyclonal expansion of T-lymphocytes. Histopathology confirmed the mass to be a thymoma, supporting the diagnosis of thymoma-associated T-cell lymphocytosis. Resolution of the lymphocytosis after removal of the thymoma provided further evidence for this diagnosis. To the authors' knowledge, this case is only the second report of thymoma-associated peripheral lymphocytosis in the veterinary literature, and is the first to report a confirmed thymoma-associated peripheral gamma delta T-cell lymphocytosis in a dog.

Monoclonal rearrangements of antigen receptor genes in lymphoproliferative diseases are characterized by the specific sequence and the length of their junctional region, which-can be used as markers of the proliferating clone. PCR techniques have greatly simplified routine detection of monoclonal rearrangements. But on the one hand, identification of the sequences requires sequencing methods and on the other hand, sizing of rearrangements by conventional analysis of PCR products on agarose or nondenaturing polyacrylamide gels may be uncertain. We have developed an approach based on amplification of rearranged IGH, TCRG and TCRD locus by fluorescent PCR associated to a computerized analysis of generated PCR products allowing their objective sizing. We tested this method on DNA samples from patients with acute 14 lymphoblastic leukemia and chronic lymphocytic leukemia, whose pattern of JGH and TCRG rearrangements had been previously identified by Southern blot techniques TCRG-PCR assay allowed detection of 100% of rearranged samples. No false-negative results were found but a high rate (60%) of Southern-negative and PCR-positive samples were identified TCRD PCR-assay detected VD1-JD1 or VD2-D2/3 rearrangements in both acute lymphoblastic leukemia and chronic lymphocytic leukemia samples. IGH PCR assay permitted detection of all known rearranged samples. The sensitivity of these three different PCR assays (1% leukemic cells) was equivalent to that of other published PCR protocols. These results show the validity and reliability of the fluorescent PCR method for routine detection of IGH, TCRG and TCRD rearrangements. Sizing of PCR products by computerized analysis was also validated. It provides additional information on rearrangement patterns in lymphoproliferative diseases, as clonal rearrangements can be recognized by their size. This can be of great interest in various circumstances particularly for detection and follow-up of oligoclonality.