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Pitfalls in TCR gene clonality testing: teaching cases.

Groenen PJ, Langerak AW, van Dongen JJ, van Krieken JH - J Hematop (2008)

Bottom Line: Several immunobiological and technical pitfalls that should be taken into account to avoid misinterpretation of data are addressed in this report.Furthermore, we discuss the need to integrate the molecular data with those from immunohistology, and preferably also flow cytometric immunophenotyping, for appropriate interpretation.Such an interactive, multidisciplinary diagnostic model guarantees integration of available data to reach the most reliable diagnosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Radboud University Nijmegen Medical Centre, Geert Grooteplein 24, 6525 GA, Nijmegen, The Netherlands, P.Groenen@pathol.umcn.nl.

ABSTRACT
Clonality testing in T-lymphoproliferations has technically become relatively easy to perform in routine laboratories using standardized multiplex polymerase chain reaction protocols for T-cell receptor (TCR) gene analysis as developed by the BIOMED-2 Concerted Action BMH4-CT98-3936. Expertise with clonality diagnostics and knowledge about the biology of TCR gene recombination are essential for correct interpretation of TCR clonality data. Several immunobiological and technical pitfalls that should be taken into account to avoid misinterpretation of data are addressed in this report. Furthermore, we discuss the need to integrate the molecular data with those from immunohistology, and preferably also flow cytometric immunophenotyping, for appropriate interpretation. Such an interactive, multidisciplinary diagnostic model guarantees integration of available data to reach the most reliable diagnosis.

No MeSH data available.


Related in: MedlinePlus

Histology of the brain biopsy, showing a mainly lymphoid infiltrate with a mixture of lymphocytes, including some larger atypical ones that are CD3 positive
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Fig7: Histology of the brain biopsy, showing a mainly lymphoid infiltrate with a mixture of lymphocytes, including some larger atypical ones that are CD3 positive

Mentions: Case 2 concerns a patient with an atypical T-cell infiltrate in the cerebellum, shown in Fig. 7. Clonality assessment was performed to assess whether infiltrated T cells were clonal, which may fit to a malignant T-cell process. FFPE tissue, containing 30% of T cells, most of which were suspected to be malignant, was available for clonality testing. Amplification of the extracted DNA was good (400 bp products could be amplified). Due to the genomic organization of the TCRG locus and the primer compositions of the BIOMED-2 TCRG multiplex tubes, the multiple (at least three) “clonal” TCRG peaks observed as shown in Fig. 8 are consistent with the presence of at least two dominant T-cell clones (one clone with TCRG biallelic rearrangements and one with a single TCRG rearrangement) in a polyclonal background of T cells. The molecular data support the morphological findings of a suspect lesion in the cerebellum, which now is classified as a peripheral T-cell lymphoma.Fig. 7


Pitfalls in TCR gene clonality testing: teaching cases.

Groenen PJ, Langerak AW, van Dongen JJ, van Krieken JH - J Hematop (2008)

Histology of the brain biopsy, showing a mainly lymphoid infiltrate with a mixture of lymphocytes, including some larger atypical ones that are CD3 positive
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2713482&req=5

Fig7: Histology of the brain biopsy, showing a mainly lymphoid infiltrate with a mixture of lymphocytes, including some larger atypical ones that are CD3 positive
Mentions: Case 2 concerns a patient with an atypical T-cell infiltrate in the cerebellum, shown in Fig. 7. Clonality assessment was performed to assess whether infiltrated T cells were clonal, which may fit to a malignant T-cell process. FFPE tissue, containing 30% of T cells, most of which were suspected to be malignant, was available for clonality testing. Amplification of the extracted DNA was good (400 bp products could be amplified). Due to the genomic organization of the TCRG locus and the primer compositions of the BIOMED-2 TCRG multiplex tubes, the multiple (at least three) “clonal” TCRG peaks observed as shown in Fig. 8 are consistent with the presence of at least two dominant T-cell clones (one clone with TCRG biallelic rearrangements and one with a single TCRG rearrangement) in a polyclonal background of T cells. The molecular data support the morphological findings of a suspect lesion in the cerebellum, which now is classified as a peripheral T-cell lymphoma.Fig. 7

Bottom Line: Several immunobiological and technical pitfalls that should be taken into account to avoid misinterpretation of data are addressed in this report.Furthermore, we discuss the need to integrate the molecular data with those from immunohistology, and preferably also flow cytometric immunophenotyping, for appropriate interpretation.Such an interactive, multidisciplinary diagnostic model guarantees integration of available data to reach the most reliable diagnosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Radboud University Nijmegen Medical Centre, Geert Grooteplein 24, 6525 GA, Nijmegen, The Netherlands, P.Groenen@pathol.umcn.nl.

ABSTRACT
Clonality testing in T-lymphoproliferations has technically become relatively easy to perform in routine laboratories using standardized multiplex polymerase chain reaction protocols for T-cell receptor (TCR) gene analysis as developed by the BIOMED-2 Concerted Action BMH4-CT98-3936. Expertise with clonality diagnostics and knowledge about the biology of TCR gene recombination are essential for correct interpretation of TCR clonality data. Several immunobiological and technical pitfalls that should be taken into account to avoid misinterpretation of data are addressed in this report. Furthermore, we discuss the need to integrate the molecular data with those from immunohistology, and preferably also flow cytometric immunophenotyping, for appropriate interpretation. Such an interactive, multidisciplinary diagnostic model guarantees integration of available data to reach the most reliable diagnosis.

No MeSH data available.


Related in: MedlinePlus