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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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Flow cytometric analyses of patient 15 with T-cell large granular lymphocyte leukemia. (a) CD8+CD4- leukemic cells. (b) CD3+/CD45RA+ leukemic cells. (c) CD2+/CD7- leukemic cells and double-stained population in the region R2 consistent with normal T lymphocytes. (d) CD5-/CD25- leukemic cells and CD5+/CD25-/+ expression on normal T lymphocytes in the region R3. (e) CD7- leukemia cells express slightly weaker levels of CD43 compared with normal CD43+higherCD7+ cells consistent with normal T lymphocytes in the region R2. (f) CD3+ population with coexistence of CD16 antigens. (g) CD3+CD56- leukemic cells. (h) CD2+CD57- leukemic cells. CD2+CD57+-reactive T lymphocytes in the region R2. (i) Leukemic cells are positive for CD3 and TCRαβ. FITC, fluorescein isothiocyanate; PE, phycoerythrin; TCR, T-cell receptor.
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Figure 2: Flow cytometric analyses of patient 15 with T-cell large granular lymphocyte leukemia. (a) CD8+CD4- leukemic cells. (b) CD3+/CD45RA+ leukemic cells. (c) CD2+/CD7- leukemic cells and double-stained population in the region R2 consistent with normal T lymphocytes. (d) CD5-/CD25- leukemic cells and CD5+/CD25-/+ expression on normal T lymphocytes in the region R3. (e) CD7- leukemia cells express slightly weaker levels of CD43 compared with normal CD43+higherCD7+ cells consistent with normal T lymphocytes in the region R2. (f) CD3+ population with coexistence of CD16 antigens. (g) CD3+CD56- leukemic cells. (h) CD2+CD57- leukemic cells. CD2+CD57+-reactive T lymphocytes in the region R2. (i) Leukemic cells are positive for CD3 and TCRαβ. FITC, fluorescein isothiocyanate; PE, phycoerythrin; TCR, T-cell receptor.

Mentions: The results of lymphocyte surface marker analysis performed in 15 patients are summarized in Table 3. The typical immunophenotype of T-LGL leukemia cells was CD45+bright, CD2+bright, CD3+bright, CD4-, CD8+bright, CD25-, and CD43+weaker. CD5 and CD7 expression was variable (bright, dim, or negative) on all or part of the T-LGL leukemia cells, whereas in 3 cases lymphocytes showed an absence of both antigens. In all studied cases, T-LGL leukemia cells expressed a slightly weaker level of CD43 as compared with normal expression of CD43+higher on T lymphocytes. Aberrant expression of CD3 was found in only 1 patient. All tested cases expressed CD16. However, 10 cases showed only partial expression of this antigen, with 20% to 95% of the T-LGL leukemia cells showing reactivity. Lack of CD56 expression was noted in 10 cases; in 2 cases, CD56 was expressed in more than 50% of the T-LGL leukemia cells. In 10 cases, 20% to 100% of the T-LGL leukemia cells showed expression of CD57, whereas 3 cases were negative. HLA-DR was expressed in all tested cases in varying percentages. TCR proteins were tested in 10 cases, 8 of them expressing TCRαβ and 2 TCRγδ (Figure 2). Patient 5 with TCRγδ protein expression had two immunophenotypically different populations of T-LGL leukemia cells and is the subject of a separate report. In analyzed cases both normal reactive peripheral blood CD57+ T lymphocytes and CD57+ T-LGL leukemia cells were found. In the former no loss of expression of any pan-T antigens was observed, in the latter typical abnormalities of pan-T-cell antigens were noted. In patients 10 and 11, with no rearrangement of the TCR genes, T-LGLs were characterized by normal expression of T-cell antigens.


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)

Flow cytometric analyses of patient 15 with T-cell large granular lymphocyte leukemia. (a) CD8+CD4- leukemic cells. (b) CD3+/CD45RA+ leukemic cells. (c) CD2+/CD7- leukemic cells and double-stained population in the region R2 consistent with normal T lymphocytes. (d) CD5-/CD25- leukemic cells and CD5+/CD25-/+ expression on normal T lymphocytes in the region R3. (e) CD7- leukemia cells express slightly weaker levels of CD43 compared with normal CD43+higherCD7+ cells consistent with normal T lymphocytes in the region R2. (f) CD3+ population with coexistence of CD16 antigens. (g) CD3+CD56- leukemic cells. (h) CD2+CD57- leukemic cells. CD2+CD57+-reactive T lymphocytes in the region R2. (i) Leukemic cells are positive for CD3 and TCRαβ. FITC, fluorescein isothiocyanate; PE, phycoerythrin; TCR, T-cell receptor.
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Related In: Results  -  Collection

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Show All Figures
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Figure 2: Flow cytometric analyses of patient 15 with T-cell large granular lymphocyte leukemia. (a) CD8+CD4- leukemic cells. (b) CD3+/CD45RA+ leukemic cells. (c) CD2+/CD7- leukemic cells and double-stained population in the region R2 consistent with normal T lymphocytes. (d) CD5-/CD25- leukemic cells and CD5+/CD25-/+ expression on normal T lymphocytes in the region R3. (e) CD7- leukemia cells express slightly weaker levels of CD43 compared with normal CD43+higherCD7+ cells consistent with normal T lymphocytes in the region R2. (f) CD3+ population with coexistence of CD16 antigens. (g) CD3+CD56- leukemic cells. (h) CD2+CD57- leukemic cells. CD2+CD57+-reactive T lymphocytes in the region R2. (i) Leukemic cells are positive for CD3 and TCRαβ. FITC, fluorescein isothiocyanate; PE, phycoerythrin; TCR, T-cell receptor.
Mentions: The results of lymphocyte surface marker analysis performed in 15 patients are summarized in Table 3. The typical immunophenotype of T-LGL leukemia cells was CD45+bright, CD2+bright, CD3+bright, CD4-, CD8+bright, CD25-, and CD43+weaker. CD5 and CD7 expression was variable (bright, dim, or negative) on all or part of the T-LGL leukemia cells, whereas in 3 cases lymphocytes showed an absence of both antigens. In all studied cases, T-LGL leukemia cells expressed a slightly weaker level of CD43 as compared with normal expression of CD43+higher on T lymphocytes. Aberrant expression of CD3 was found in only 1 patient. All tested cases expressed CD16. However, 10 cases showed only partial expression of this antigen, with 20% to 95% of the T-LGL leukemia cells showing reactivity. Lack of CD56 expression was noted in 10 cases; in 2 cases, CD56 was expressed in more than 50% of the T-LGL leukemia cells. In 10 cases, 20% to 100% of the T-LGL leukemia cells showed expression of CD57, whereas 3 cases were negative. HLA-DR was expressed in all tested cases in varying percentages. TCR proteins were tested in 10 cases, 8 of them expressing TCRαβ and 2 TCRγδ (Figure 2). Patient 5 with TCRγδ protein expression had two immunophenotypically different populations of T-LGL leukemia cells and is the subject of a separate report. In analyzed cases both normal reactive peripheral blood CD57+ T lymphocytes and CD57+ T-LGL leukemia cells were found. In the former no loss of expression of any pan-T antigens was observed, in the latter typical abnormalities of pan-T-cell antigens were noted. In patients 10 and 11, with no rearrangement of the TCR genes, T-LGLs were characterized by normal expression of T-cell antigens.

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