Characterization of feline T cell receptor gamma (TCRG) variable region genes for the molecular diagnosis of feline intestinal T cell lymphoma.
A diagnosis of intestinal lymphoma is currently made on the basis of clinical and morphologic criteria. This can prove problematic for many reasons that include inadequate sample size, the coexistence of lymphoma and inflammation, and the inability to assess architectural integrity of all tissue compartments in biopsy specimens obtained endoscopically. The detection of a clonal population of cells in a lymphoproliferative lesion represents an important criterion for the diagnosis of neoplasia, but this has not been assessed in feline intestinal lymphoma. T cell receptor gamma (TCRG) gene rearrangement analysis using polymerase chain reaction (PCR) is a methodology that can be used to detect clonality in T cell populations. The basis of this assay depends on the assessment of the junctional diversity that results from rearrangement of TCRG V (variable) and J (joining) gene segments. Feline TCRG transcripts from normal small intestine and spleen were obtained using a rapid amplification of cDNA ends (5'RACE) method. Limited diversity of TCRG V and J gene segments was observed. The high degree of sequence homology in the TCRG V and J gene segments was exploited to develop a PCR test for the assessment of TCRG V--J junctional diversity and hence clonality determination of T cell populations in cats. Molecular clonality determination was applied to feline intestinal lymphoplasmacytic inflammatory bowel disease (IBD) (9 cats), and transmural and mucosal T cell lymphoma (28 cats). Clonal rearrangement of the TCRG V--J junction was detected in 22 of 28 intestinal T cell lymphomas, and oligoclonality was detected in 3 intestinal T cell lymphomas. This contrasted with the detection of polyclonal rearrangement in normal intestinal tissues (3 cats) and in lymphoplasmacytic IBD (9 cats). It is proposed that assessment of TCRG V--J junctional diversity for the detection of clonality represents an important adjunctive tool for the diagnosis of T cell lymphoma in the cat.
Moore
Peter F
PF
Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616, USA. pfmoore@ucdavis.edu
Woo
Jennifer C
JC
Vernau
William
W
Kosten
Sandra
S
Graham
Petra S
PS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002371:Cat Diseases / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology; D002415:Cats; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D020180:Heteroduplex Analysis / Q000662:veterinary; D015212:Inflammatory Bowel Diseases / Q000175:diagnosis / Q000235:genetics / Q000276:immunology / Q000662:veterinary; D007414:Intestinal Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D016399:Lymphoma, T-Cell / Q000175:diagnosis / Q000235:genetics / Q000276:immunology / Q000662:veterinary*; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D017386:Sequence Homology, Amino Acid; D014410:Tumor Stem Cell Assay / Q000662:veterinary
D018076:DNA, Complementary; D004273:DNA, Neoplasm