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Characteristics of T-cell large granular lymphocyte proliferations associated with neutropenia and inflammatory arthropathy.

Prochorec-Sobieszek M, Rymkiewicz G, Makuch-Łasica H, Majewski M, Michalak K, Rupiński R, Warzocha K, Maryniak R - Arthritis Res. Ther. (2008)

Bottom Line: Bone marrow trephines from T-LGL leukemia patients showed interstitial clusters and intrasinusoidal linear infiltrations of CD3+/CD8+/CD57+/granzyme B+ lymphocytes, reactive lymphoid nodules, and decreased or normal granulocyte precursor count with left-shifted maturation.RA and neutropenia patients represented a continuous spectrum of T-LGL proliferations, although monoclonal expansions were most frequently observed.The histopathological pattern and immunophenotype of bone marrow infiltration as well as molecular characteristics were similar in T-LGL leukemia patients with and without arthritis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathomorphology, Institute of Hematology and Transfusion Medicine, I, Gandhi 14, 02-776 Warsaw, Poland. monika.prochorec@interia.pl

ABSTRACT

Introduction: The purpose of this study was to analyze the data of patients with T-cell large granular lymphocyte (T-LGL) lymphocytosis associated with inflammatory arthropathy or with no arthritis symptoms.

Methods: Clinical, serological as well as histopathological, immunohistochemical, and flow cytometric evaluations of blood/bone marrow of 21 patients with T-LGL lymphocytosis were performed. The bone marrow samples were also investigated for T-cell receptor (TCR) and immunoglobulin (IG) gene rearrangements by polymerase chain reaction with heteroduplex analysis.

Results: Neutropenia was observed in 21 patients, splenomegaly in 10, autoimmune diseases such as rheumatoid arthritis (RA) in 9, unclassified arthritis resembling RA in 2, and autoimmune thyroiditis in 5 patients. T-LGL leukemia was recognized in 19 cases. Features of Felty syndrome were observed in all RA patients, representing a spectrum of T-LGL proliferations from reactive polyclonal through transitional between reactive and monoclonal to T-LGL leukemia. Bone marrow trephines from T-LGL leukemia patients showed interstitial clusters and intrasinusoidal linear infiltrations of CD3+/CD8+/CD57+/granzyme B+ lymphocytes, reactive lymphoid nodules, and decreased or normal granulocyte precursor count with left-shifted maturation. In three-color flow cytometry (FCM), T-LGL leukemia cells demonstrated CD2, CD3, and CD8 expression as well as a combination of CD16, CD56, or CD57. Abnormalities of other T-cell antigen expressions (especially CD5, CD7, and CD43) were also detected. In patients with polyclonal T-LGL lymphocytosis, T cells were dispersed in the bone marrow and the expression of pan-T-cell antigens in FCM was normal. Molecular studies revealed TCRB and TCRG gene rearrangements in 13 patients and TCRB, TCRG, and TCRD in 4 patients. The most frequently rearranged regions of variable genes were Vbeta-Jbeta1, Jbeta2 and Vgamma If Vgamma10-Jgamma. Moreover, in 4 patients, additional rearrangements of IG kappa and lambda variable genes of B cells were also observed.

Conclusion: RA and neutropenia patients represented a continuous spectrum of T-LGL proliferations, although monoclonal expansions were most frequently observed. The histopathological pattern and immunophenotype of bone marrow infiltration as well as molecular characteristics were similar in T-LGL leukemia patients with and without arthritis.

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Ethidium bromide-stained polyacrylamide gel showing polymerase chain reaction products derived from TCR gene rearrangements in patients with rheumatoid arthritis and T-cell large granular lymphocyte (T-LGL) proliferations. (a) Polyclonal expansion of T-LGLs in patient 10. Lane 1: TCRB gene rearrangement–negative, polyclonal smear (tube A); lane 2: TCRB gene rearrangement-negative, polyclonal smear (tube B); lane 3: TCRB gene rearrangement-negative, polyclonal smear (tube C); lane 4: standard 50 base pairs (bp); lane 5: TCRG gene rearrangement-negative, polyclonal smear (tube A); lane 6: TCRG gene rearrangement-negative, polyclonal smear (tube B); and lane 7: TCRD gene rearrangement-negative, polyclonal smear. (b) Monoclonal expansion in polyclonal background in patient 7. Lane 1: TCRG gene rearrangement: monoclonal product 180 bp (i) in tube A; lane 2: TCRG gene rearrangement: monoclonal product 210 bp (ii) in polyclonal background (tube B); lane 3: TCRD gene rearrangement: monoclonal product 160 bp (iii); lane 4: TCRB (tube A) gene rearrangement-negative, polyclonal smear; lane 5: standard 50 bp; lane 6: TCRB (tube B) gene rearrangement-negative, polyclonal smear; and lane 7: TCRB (tube C) gene rearrangement-negative, polyclonal smear. (c) Monoclonal gene rearrangements in patient 1 with T-LGL leukemia. Lane 1: TCRB gene rearrangement-negative (tube A); lane 2: TCRB gene rearrangement-positive, monoclonal product 250 bp (iv) in tube B; lane 3: TCRB (tube C): gene rearrangement-negative, polyclonal smear; lane 4: standard 50 bp; lane 5: TCRG gene rearrangement-positive, monoclonal product 230 bp (v) in tube A; lane 6: TCRG gene rearrangement-positive, monoclonal product 180 bp (vi) in tube B; and lane 7: TCRD gene rearrangement-negative, polyclonal smear. TCR, T-cell receptor.
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Figure 3: Ethidium bromide-stained polyacrylamide gel showing polymerase chain reaction products derived from TCR gene rearrangements in patients with rheumatoid arthritis and T-cell large granular lymphocyte (T-LGL) proliferations. (a) Polyclonal expansion of T-LGLs in patient 10. Lane 1: TCRB gene rearrangement–negative, polyclonal smear (tube A); lane 2: TCRB gene rearrangement-negative, polyclonal smear (tube B); lane 3: TCRB gene rearrangement-negative, polyclonal smear (tube C); lane 4: standard 50 base pairs (bp); lane 5: TCRG gene rearrangement-negative, polyclonal smear (tube A); lane 6: TCRG gene rearrangement-negative, polyclonal smear (tube B); and lane 7: TCRD gene rearrangement-negative, polyclonal smear. (b) Monoclonal expansion in polyclonal background in patient 7. Lane 1: TCRG gene rearrangement: monoclonal product 180 bp (i) in tube A; lane 2: TCRG gene rearrangement: monoclonal product 210 bp (ii) in polyclonal background (tube B); lane 3: TCRD gene rearrangement: monoclonal product 160 bp (iii); lane 4: TCRB (tube A) gene rearrangement-negative, polyclonal smear; lane 5: standard 50 bp; lane 6: TCRB (tube B) gene rearrangement-negative, polyclonal smear; and lane 7: TCRB (tube C) gene rearrangement-negative, polyclonal smear. (c) Monoclonal gene rearrangements in patient 1 with T-LGL leukemia. Lane 1: TCRB gene rearrangement-negative (tube A); lane 2: TCRB gene rearrangement-positive, monoclonal product 250 bp (iv) in tube B; lane 3: TCRB (tube C): gene rearrangement-negative, polyclonal smear; lane 4: standard 50 bp; lane 5: TCRG gene rearrangement-positive, monoclonal product 230 bp (v) in tube A; lane 6: TCRG gene rearrangement-positive, monoclonal product 180 bp (vi) in tube B; and lane 7: TCRD gene rearrangement-negative, polyclonal smear. TCR, T-cell receptor.

Mentions: The detailed results of TCR gene rearrangement tests are summarized in Table 2. Two patients (patients 10 and 11) had no rearrangement in TCR genes consistent with polyclonal lymphocytosis (Figure 3a). Patient 7, apart from monoclonal TCR rearrangement in the delta chain, showed a weak monoclonal product in the TCRG Vγ9, Vγ11-Jγ region in polyclonal background (Figure 3b). Clearly monoclonal TCR gene rearrangements were detected in 16 patients (Figure 3c). Three patients had rearrangements in genes coding beta chains, 10 showed clonality in genes coding beta and gamma chains, and 3 had rearrangement in genes coding beta, gamma, and delta chains. The spectrum of TCR gene rearrangements was quite variable, although there were some repeated uses observed. The most frequently rearranged regions of variable genes were Vβ-Jβ1, Jβ2 (13 patients) and Vγ If Vγ10-Jγ (13 patients). Moreover, 4 patients (patients 4, 7, 9, and 15) also presented rearrangements of immunoglobulin kappa variable (IGKV) or lambda variable (IGLV) genes. Classic cytogenetic tests with GTG banding of the bone marrow cells were performed in 7 patients (patients 1, 4, 9, 12, 13, 15, and 18) and revealed a normal karyotype.


Characteristics of T-cell large granular lymphocyte proliferations associated with neutropenia and inflammatory arthropathy.

Prochorec-Sobieszek M, Rymkiewicz G, Makuch-Łasica H, Majewski M, Michalak K, Rupiński R, Warzocha K, Maryniak R - Arthritis Res. Ther. (2008)

Ethidium bromide-stained polyacrylamide gel showing polymerase chain reaction products derived from TCR gene rearrangements in patients with rheumatoid arthritis and T-cell large granular lymphocyte (T-LGL) proliferations. (a) Polyclonal expansion of T-LGLs in patient 10. Lane 1: TCRB gene rearrangement–negative, polyclonal smear (tube A); lane 2: TCRB gene rearrangement-negative, polyclonal smear (tube B); lane 3: TCRB gene rearrangement-negative, polyclonal smear (tube C); lane 4: standard 50 base pairs (bp); lane 5: TCRG gene rearrangement-negative, polyclonal smear (tube A); lane 6: TCRG gene rearrangement-negative, polyclonal smear (tube B); and lane 7: TCRD gene rearrangement-negative, polyclonal smear. (b) Monoclonal expansion in polyclonal background in patient 7. Lane 1: TCRG gene rearrangement: monoclonal product 180 bp (i) in tube A; lane 2: TCRG gene rearrangement: monoclonal product 210 bp (ii) in polyclonal background (tube B); lane 3: TCRD gene rearrangement: monoclonal product 160 bp (iii); lane 4: TCRB (tube A) gene rearrangement-negative, polyclonal smear; lane 5: standard 50 bp; lane 6: TCRB (tube B) gene rearrangement-negative, polyclonal smear; and lane 7: TCRB (tube C) gene rearrangement-negative, polyclonal smear. (c) Monoclonal gene rearrangements in patient 1 with T-LGL leukemia. Lane 1: TCRB gene rearrangement-negative (tube A); lane 2: TCRB gene rearrangement-positive, monoclonal product 250 bp (iv) in tube B; lane 3: TCRB (tube C): gene rearrangement-negative, polyclonal smear; lane 4: standard 50 bp; lane 5: TCRG gene rearrangement-positive, monoclonal product 230 bp (v) in tube A; lane 6: TCRG gene rearrangement-positive, monoclonal product 180 bp (vi) in tube B; and lane 7: TCRD gene rearrangement-negative, polyclonal smear. TCR, T-cell receptor.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2483444&req=5

Figure 3: Ethidium bromide-stained polyacrylamide gel showing polymerase chain reaction products derived from TCR gene rearrangements in patients with rheumatoid arthritis and T-cell large granular lymphocyte (T-LGL) proliferations. (a) Polyclonal expansion of T-LGLs in patient 10. Lane 1: TCRB gene rearrangement–negative, polyclonal smear (tube A); lane 2: TCRB gene rearrangement-negative, polyclonal smear (tube B); lane 3: TCRB gene rearrangement-negative, polyclonal smear (tube C); lane 4: standard 50 base pairs (bp); lane 5: TCRG gene rearrangement-negative, polyclonal smear (tube A); lane 6: TCRG gene rearrangement-negative, polyclonal smear (tube B); and lane 7: TCRD gene rearrangement-negative, polyclonal smear. (b) Monoclonal expansion in polyclonal background in patient 7. Lane 1: TCRG gene rearrangement: monoclonal product 180 bp (i) in tube A; lane 2: TCRG gene rearrangement: monoclonal product 210 bp (ii) in polyclonal background (tube B); lane 3: TCRD gene rearrangement: monoclonal product 160 bp (iii); lane 4: TCRB (tube A) gene rearrangement-negative, polyclonal smear; lane 5: standard 50 bp; lane 6: TCRB (tube B) gene rearrangement-negative, polyclonal smear; and lane 7: TCRB (tube C) gene rearrangement-negative, polyclonal smear. (c) Monoclonal gene rearrangements in patient 1 with T-LGL leukemia. Lane 1: TCRB gene rearrangement-negative (tube A); lane 2: TCRB gene rearrangement-positive, monoclonal product 250 bp (iv) in tube B; lane 3: TCRB (tube C): gene rearrangement-negative, polyclonal smear; lane 4: standard 50 bp; lane 5: TCRG gene rearrangement-positive, monoclonal product 230 bp (v) in tube A; lane 6: TCRG gene rearrangement-positive, monoclonal product 180 bp (vi) in tube B; and lane 7: TCRD gene rearrangement-negative, polyclonal smear. TCR, T-cell receptor.
Mentions: The detailed results of TCR gene rearrangement tests are summarized in Table 2. Two patients (patients 10 and 11) had no rearrangement in TCR genes consistent with polyclonal lymphocytosis (Figure 3a). Patient 7, apart from monoclonal TCR rearrangement in the delta chain, showed a weak monoclonal product in the TCRG Vγ9, Vγ11-Jγ region in polyclonal background (Figure 3b). Clearly monoclonal TCR gene rearrangements were detected in 16 patients (Figure 3c). Three patients had rearrangements in genes coding beta chains, 10 showed clonality in genes coding beta and gamma chains, and 3 had rearrangement in genes coding beta, gamma, and delta chains. The spectrum of TCR gene rearrangements was quite variable, although there were some repeated uses observed. The most frequently rearranged regions of variable genes were Vβ-Jβ1, Jβ2 (13 patients) and Vγ If Vγ10-Jγ (13 patients). Moreover, 4 patients (patients 4, 7, 9, and 15) also presented rearrangements of immunoglobulin kappa variable (IGKV) or lambda variable (IGLV) genes. Classic cytogenetic tests with GTG banding of the bone marrow cells were performed in 7 patients (patients 1, 4, 9, 12, 13, 15, and 18) and revealed a normal karyotype.

Bottom Line: Bone marrow trephines from T-LGL leukemia patients showed interstitial clusters and intrasinusoidal linear infiltrations of CD3+/CD8+/CD57+/granzyme B+ lymphocytes, reactive lymphoid nodules, and decreased or normal granulocyte precursor count with left-shifted maturation.RA and neutropenia patients represented a continuous spectrum of T-LGL proliferations, although monoclonal expansions were most frequently observed.The histopathological pattern and immunophenotype of bone marrow infiltration as well as molecular characteristics were similar in T-LGL leukemia patients with and without arthritis.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathomorphology, Institute of Hematology and Transfusion Medicine, I, Gandhi 14, 02-776 Warsaw, Poland. monika.prochorec@interia.pl

ABSTRACT

Introduction: The purpose of this study was to analyze the data of patients with T-cell large granular lymphocyte (T-LGL) lymphocytosis associated with inflammatory arthropathy or with no arthritis symptoms.

Methods: Clinical, serological as well as histopathological, immunohistochemical, and flow cytometric evaluations of blood/bone marrow of 21 patients with T-LGL lymphocytosis were performed. The bone marrow samples were also investigated for T-cell receptor (TCR) and immunoglobulin (IG) gene rearrangements by polymerase chain reaction with heteroduplex analysis.

Results: Neutropenia was observed in 21 patients, splenomegaly in 10, autoimmune diseases such as rheumatoid arthritis (RA) in 9, unclassified arthritis resembling RA in 2, and autoimmune thyroiditis in 5 patients. T-LGL leukemia was recognized in 19 cases. Features of Felty syndrome were observed in all RA patients, representing a spectrum of T-LGL proliferations from reactive polyclonal through transitional between reactive and monoclonal to T-LGL leukemia. Bone marrow trephines from T-LGL leukemia patients showed interstitial clusters and intrasinusoidal linear infiltrations of CD3+/CD8+/CD57+/granzyme B+ lymphocytes, reactive lymphoid nodules, and decreased or normal granulocyte precursor count with left-shifted maturation. In three-color flow cytometry (FCM), T-LGL leukemia cells demonstrated CD2, CD3, and CD8 expression as well as a combination of CD16, CD56, or CD57. Abnormalities of other T-cell antigen expressions (especially CD5, CD7, and CD43) were also detected. In patients with polyclonal T-LGL lymphocytosis, T cells were dispersed in the bone marrow and the expression of pan-T-cell antigens in FCM was normal. Molecular studies revealed TCRB and TCRG gene rearrangements in 13 patients and TCRB, TCRG, and TCRD in 4 patients. The most frequently rearranged regions of variable genes were Vbeta-Jbeta1, Jbeta2 and Vgamma If Vgamma10-Jgamma. Moreover, in 4 patients, additional rearrangements of IG kappa and lambda variable genes of B cells were also observed.

Conclusion: RA and neutropenia patients represented a continuous spectrum of T-LGL proliferations, although monoclonal expansions were most frequently observed. The histopathological pattern and immunophenotype of bone marrow infiltration as well as molecular characteristics were similar in T-LGL leukemia patients with and without arthritis.

Show MeSH
Related in: MedlinePlus