title: Polyclonal expansion of T-cell receptor-gamma delta+ T lymphocytes associated with neutropenia and thrombocytopenia.
abstract: A patient with large granular lymphocyte (LGL) expansion (T-gamma lymphocytosis), neutropenia and thrombocytopenia was studied longitudinally. Analysis of peripheral blood mononuclear cells (PBMC) demonstrated an unusual large proportion of CD3+ T-lymphocytes expressing a gamma delta T-cell receptor (TcR-gamma delta). Immunofluorescence (IF) stainings with subset-specific monoclonal antibodies showed a fluctuating expansion of TcR-gamma delta+ T-cells expressing V gamma 9 and V delta 2 variable (V) gene segments. Biochemical characterization of PBMC showed the presence of a disulphide-linked TcR-gamma delta. TcR gene rearrangement studies on sorted TcR-gamma delta+ T-cells showed rearrangements of V gamma 9-J gamma 1.2 and V delta 2-J delta 1 V and joining (J) gene segments, thereby confirming the IF staining results. These data alone did not allow us to determine whether the TcR-gamma delta+ LGL expansion represented a polyclonal or monoclonal proliferation, because the combinatorial repertoire of TcR-gamma delta receptors is limited due to the availability of only a few V and J segments within the TcR-gamma and TcR-delta genes and because of the preferential usage of V gamma 9-J gamma 1.2 and V delta 2-J delta 1 rearrangements by TcR-gamma delta+ T-cells in blood of healthy individuals. We therefore used polymerase chain reaction (PCR)-mediated amplification of the TcR-gamma delta rearrangements, using specific V gamma 9, J gamma 1.2, V delta 2, and J delta 1 oligonucleotides to determine the junctional diversity of the TcR-gamma delta+ T-cell population. Sequence analysis of the PCR products obtained revealed a mixture of different junctional region sequences compatible with a polyclonal expansion. This is in contrast to the few reported TcR-gamma delta+ LGL and the majority of TcR-alpha beta+ LGL expansions, which appeared to consist of monoclonal proliferations.
authors.affiliation: Division of Immunology, The Netherlands Cancer Institute, Amsterdam.
authors.forename: J W
authors.initials: JW
authors.lastname: van Oostveen
authors.forename: T M
authors.initials: TM
authors.lastname: Breit
authors.forename: J T
authors.initials: JT
authors.lastname: de Wolf
authors.forename: R M
authors.initials: RM
authors.lastname: Brandt
authors.forename: J W
authors.initials: JW
authors.lastname: Smit
authors.forename: J J
authors.initials: JJ
authors.lastname: van Dongen
authors.forename: J
authors.initials: J
authors.lastname: Borst
authors.forename: C J
authors.initials: CJ
authors.lastname: Melief
mesh_terms: D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis / Q000235:genetics; D015139:Blotting, Southern; D017252:CD3 Complex; D006801:Humans; D016130:Immunophenotyping; D007958:Leukocyte Count; D008137:Longitudinal Studies; D008297:Male; D008875:Middle Aged; D009503:Neutropenia / Q000150:complications / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011948:Receptors, Antigen, T-Cell / Q000032:analysis / Q000235:genetics; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000032:analysis* / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*; D013921:Thrombocytopenia / Q000150:complications / Q000276:immunology*
chemical_list: D000945:Antigens, Differentiation, T-Lymphocyte; D017252:CD3 Complex; D011948:Receptors, Antigen, T-Cell; D016692:Receptors, Antigen, T-Cell, gamma-delta