Immunophenotyping and Gene Rearrangement Analysis in Dogs with Lymphoproliferative Disorders Characterized by Small-Cell Lymphocytosis

Yagihara, Hiroko; Uematsu, Yosuke; Koike, Ayumi; Tamura, Kyoichi; Isotani, Mayu; Yamaguchi, Takahiro; Ono, Kenichiro; Washizu, Tsukimi; Bonkobara, Makoto

doi:10.1177/104063870902100203

Cited by 13 publications

(11 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The positive result was confirmed with repeat testing at CSU. The sensitivity of our assay falls within that reported for similar primer sets in other studies (5–7, 10, 20). …”

Section: Methodssupporting

confidence: 88%

“…We found a similar result in this study, in which tumor cells in dogs with AML expressed B or T cell surface antigens, yet displayed non-clonal, biclonal, or B or T cell receptor clonal rearrangements. Although non-clonal or polyclonal results on clonality testing have been used to support a diagnosis of AML in previous reports (6, 10, 14, 15), false-negative reactions for clonality do occur in lymphoid neoplasms, with reported sensitivities ranging from 67 to 98% (5–7, 10, 20), using similar primer sets to that used herein.…”

Section: Discussionmentioning

confidence: 92%

“…Recently, Keller and colleagues have also advocated against the use of clonality testing for distinguishing B from T cell neoplasms (1). However, in many cases of lymphoid neoplasia, the tumor shows fidelity between clonality and other phenotyping tests (IHC or flow cytometry) (2, 6, 20), and clonality assessment may still be useful, at the very least for confirming neoplasia, in sites where sufficient cells cannot be retrieved for other phenotyping techniques or invasive diagnostic procedures are contraindicated (12). …”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Dogs with Acute Myeloid Leukemia Have Clonal Rearrangements in T and B Cell Receptors

et al. 2017

View full text Add to dashboard Cite

Clonality testing for rearrangements in the complementarity-determining region 3 of the immunoglobulin heavy chain of B lymphocytes (B cell receptor) and the T cell receptor of T lymphocytes helps distinguish between clonal and non-clonal expansions of lymphocytes. There are rare reports of clonally rearranged T and B cell receptors in dogs with acute myeloid leukemia (AML). Our objective was to determine the frequency of clonally rearranged T and B cell receptors in dogs with AML. Archived slides from historical cases of AML (from January 2010 to June 2013) and slides or liquid specimens [blood, bone marrow (BM), body cavity fluid, or tissue aspirates] from cases of AML diagnosed between June 2013 and February 2017 were used in the study. A diagnosis of AML was made on the basis of more than 20% immature neoplastic cells (“blasts”) in blood, BM, or extramedullary tissues, displaying features of myeloid differentiation. Myeloid differentiation was based on a combination of morphologic criteria, positive flow cytometric labeling for surface antigens typical of myeloid origin (e.g., CD11b, CD11c, CD14 with a general lack of expression of T or B cell markers), or positive cytochemical staining reactions for myeloid-associated enzymes (e.g., alkaline phosphatase, chloroacetate esterase). There were 63 cases of AML diagnosed during this period; however, slides or liquid specimens with sufficient DNA for testing were only obtained from 25 dogs. Affected dogs represented various breeds and were a median of 8 years old, with more male (64%) than female (36%) dogs. Common clinical signs were peripheral or internal lymphadenopathy (10/25 dogs, 40%) and hepatomegaly or splenomegaly (10/25 dogs combined, 40%). Typical hematologic findings were bi- or pancytopenia (23/25 dogs, 92%), with circulating blasts (21/25, 84%). Solitary clonal (4 B cell, 6 T cell) and biclonal (6 B and T cell) rearrangements in B or T cell receptors were found in 16 dogs (64%). Our results indicate that dogs with AML can have a high frequency of clonally rearranged T or B cell receptors, including biclonality, and clonality testing should not be used as a tool to distinguish between acute leukemia of myeloid or lymphoid origin.

show abstract

“…The positive result was confirmed with repeat testing at CSU. The sensitivity of our assay falls within that reported for similar primer sets in other studies (5–7, 10, 20). …”

Section: Methodssupporting

confidence: 88%

Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Dogs with Acute Myeloid Leukemia Have Clonal Rearrangements in T and B Cell Receptors

et al. 2017

View full text Add to dashboard Cite

show abstract

“…1 Polymerase chain reaction (PCR) for antigen receptor rearrangement (PARR) is a method used for the detection of clonal lymphocyte populations, phenotyping of lymphocytes, detection of malignant lymphocytes in blood, and diagnosis of small-cell lymphocytosis. 2,13 It is reportedly useful for the diagnosis of canine alimentary lymphoma. 4,7 This technique is highly sensitive but tends to produce false-positive results, a phenomenon termed pseudoclonality, possibly because a single copy is not amplified during PCR because of the immunologic diversity of the target genes: immunoglobulin heavy chain (IgH) and T-cell receptor gamma (TCRc).…”

Section: <!?Show "Fnote_aff1"$^!"content-markup(/author-grp[1]/aff|/author-grp[1]/dept-list)>mentioning

confidence: 99%

Heteroduplex Polymerase Chain Reaction is Essential for Canine Receptor Rearrangement Analysis

Takanosu

Tadika

Kobayashi³

2010

J VET Diagn Invest

View full text Add to dashboard Cite

Abstract. Polymerase chain reaction (PCR) for antigen receptor rearrangement is a sensitive technique for detecting lymphocyte-proliferative disorders, but it tends to produce false-positive results, a phenomenon termed pseudoclonality. Heteroduplex analysis, which is useful to distinguish clonal reactions from pseudoclonal ones in dogs, can be applied to avoid misdiagnosis and determine the reliability of results. In the current study, PCR for antigen receptor rearrangement was used to identify clonal proliferation of lymphocytes in duodenal and lymphoid tissue from dogs presenting with chronic vomiting and enlarged peripheral lymph nodes typical of multicentric lymphoma, and the test results were verified with heteroduplex analysis. In the case of almost all of the duodenal samples, even without a histologic diagnosis of lymphoma, a distinct band similar to that observed in the case of lymphoma was obtained for both B-and T-cell clonality. All of the bands obtained from the nonneoplastic duodenum disappeared following heteroduplex analysis of the PCR product, whereas the distinct bands from the lymphoma remained. In the lymph node samples, the pseudoclonal bands that disappeared in the heteroduplex analysis were detected mainly in B cells. In conclusion, heteroduplex analysis with PCR for antigen receptor rearrangement is a suitable tool for diagnosing canine lymphoma and decreasing the possibility of misdiagnosis of pseudoclonality.

show abstract

“…55 Based on more recent studies that have identified additional less commonly expressed rearrangements and provided more detailed characterization of the complete canine TRC locus, 56,76,149 a new multiplex PARR assay has been developed for assessment of canine T-cell proliferations that has been shown to have an improved sensitivity. 55 In addition to using multiplex primers, the sensitivity of the PARR assay has also been improved by using capillary gel electrophoresis.…”

Section: Pcr To Detect Antigen Receptor Rearrangementsmentioning

confidence: 99%

Beyond H&E

et al. 2013

View full text Add to dashboard Cite

Veterinary pathology of infectious, particularly viral, and neoplastic diseases has advanced significantly with the advent of newer molecular methodologies that can detect nucleic acid of infectious agents within microscopic lesions, differentiate neoplastic from nonneoplastic cells, or determine the suitability of a targeted therapy by detecting specific mutations in certain cancers. Polymerase chain reaction-based amplification of DNA or RNA and in situ hybridization are currently the most commonly used methods for nucleic acid detection. In contrast, the main methodology used for protein detection within microscopic lesions is immunohistochemistry. Other methods that allow for analysis of nucleic acids within a particular cell type or individual cells, such as laser capture microdissection, are also available in some laboratories. This review gives an overview of the factors that influence the accurate analysis of nucleic acids in formalin-fixed tissues, as well as of different approaches to detect such targets. Keywords cancer, ISH, IHC, nucleic acid analysis, molecular pathology, PCR, virology Technical ConsiderationsTissue Handling, Fixation, and Embedding Fixation of tissues in 10% neutral buffered formalin, followed by paraffin embedding (FFPE), has long been the standard approach of tissue preparation for microscopic evaluation. The chemical principle of formalin fixation is based on the crosslinking of proteins. Formalin has been used extensively because it is inexpensive and excellently preserves morphological detail in histological sections. However, the relatively recent development and increasing use of both protein and nucleic acid detection methods have revealed some significant shortcomings associated with the formalin-based fixation process. The effects of formalin fixation on DNA are chemical modification, DNA trapping, and potentially DNA fragmentation. Chemical modification, considered the main effect, results from the direct cross-linking of proteins to DNA. 68 In addition, extensive protein-protein cross-linking and the formation of protein networks result in DNA trapping. The combination of these effects makes it much more difficult to extract DNA from FFPE tissue than from unfixed tissue, especially when methods used for extraction from fresh tissues are used on FFPE tissue without any modifications. 60,83 In addition to interfering with the ability to extract nucleic acid, formalin fixation also causes DNA fragmentation, especially when the formalin used is inadequately buffered, which then leads to formic acid formation during the fixation process. This, in turn, results in depurination of the DNA, which then becomes susceptible to cleavage by hydroxyl ions and results in the formation of single-stranded breaks.60 Similar to DNA, RNA is also degraded and chemically modified as the result of formalin fixation. The most important chemical effect of formalin on RNA is the formation of monomethylol groups with purine bases. 77 This has a significant effect on reverse transcription ...

show abstract

Immunophenotyping and Gene Rearrangement Analysis in Dogs with Lymphoproliferative Disorders Characterized by Small-Cell Lymphocytosis

Cited by 13 publications

References 16 publications

Dogs with Acute Myeloid Leukemia Have Clonal Rearrangements in T and B Cell Receptors

Dogs with Acute Myeloid Leukemia Have Clonal Rearrangements in T and B Cell Receptors

Heteroduplex Polymerase Chain Reaction is Essential for Canine Receptor Rearrangement Analysis

Beyond H&E

Contact Info

Product

Resources

About