Immunophenotypic analysis of adult patients with T-cell lymphoblastic lymphoma treated with hyper-CVAD

<p><b>Objectives:</b> Immunophenotype is an important prognostic factor for childhood and adult T-cell acute lymphoblastic leukemia. However, immunophenotypic data from adult patients with T-cell lymphoblastic lymphoma (T-LBL) are scarcely available.</p> <p><b>Methods:</b> Subjects were unselected adult patients with T-LBL who were treated with intensive chemotherapy. Immunophenotyping of tumor cells was performed according to standard techniques.</p> <p><b>Results:</b> A total of eight patients with a median age of 31 years were analyzed who received hyper-CVAD treatment for LBL. Immunophenotypic analysis showed that the most common tumor type was cortical T-cell type [early T (<i>n</i> = 2), cortical T (<i>n</i> = 4), and medullary T (<i>n</i> = 2)]. Two patients diagnosed with early T-cell type had early disease progression.</p> <p><b>Conclusions:</b> Assessment of T-cell differentiation stages in malignant T lymphoblasts would be important in choosing treatment strategies for adult patients with T-LBL.</p

Samples with <i>A20</i> deletions and/or the absence of A20 by immunohistochemistry.

<p>Abbreviations: PAL, pyothorax-associated lymphoma; NKTL, NK/T cell lymphoma, nasal type; DLBCL-e, EBV positive diffuse large B<i>-</i>cell lymphoma of the elderly; MTX-LPD, methotrexate-related lymphoproliferative disorders; LOH, loss of heterozygosity; EBER1, Epstein-Barr virus encoded RNA1; LMP-1, latent membrane protein-1; EBNA-2, EBV nuclear antigen-2; +, positive (50% or more); p+, intermediate expression (less than 50%); −, negative (0%); +/−, equivocal positive; u.d., undetermined.</p

Mono- and Bi-allelic deletions of <i>A20</i> as determined by fluorescent <i>in situ</i> hybridization.

<p>Abbreviations: PAL, pyothorax-associated lymphoma; NKTL, NK/T cell lymphoma, nasal type; DLBCL-e, EBV positive diffuse large B<i>-</i>cell lymphoma of the elderly; MTX-LPD, methotrexate-related lymphoproliferative disorders.</p

Incidence of <i>A20</i> deletion and latent membrane protein-1 (LMP-1) status.

<p>Abbreviations: PAL, pyothorax-associated lymphoma; NKTL, NK/T cell lymphoma, nasal type; DLBCL-e, EBV positive diffuse large B<i>-</i>cell lymphoma of the elderly; MTX-LPD, methotrexate-related lymphoproliferative disorders; LMP-1: +, positive (50% or more); +/−, intermediate expression (less than 50%); −, negative (0%).</p

A20 monoallelic deletion in pyothorax-associated lymphoma.

<p>(A) Diffuse proliferation of lymphoid cells, (hematoxylin-eosin stain, Olympus BX51, magnification ×200; inset ×400). (B) Positive signals in the nucleus of almost all tumor cells, (Epstein-Barr virus encoded RNA1, Olympus BX51, magnification ×400). (C) Positive signals in >50% of the tumor cells, (latent membrane protein-1, Olympus BX51, magnification ×400). (D) Monoallelic deletion of A20 detected by fluorescent <i>in situ</i> hybridization. A20 probe (orange) and chromosome 6 centromeric probe (green) (Olympus IX71, colors corrected after acquisition with Adobe Photoshop).</p

Frequency of literature-reported <i>A20</i> deletions by fluorescent <i>in situ</i> hybridization.

<p>Abbreviations: EMZL, extranodal marginal zone lymphoma; NMZL, nodal marginal zone lymphoma; SMZL, splenic marginal zone lymphoma; DLBCL-GI, gastrointestinal diffuse large B-cell lymphoma; ARL, AIDS-related lymphoma.</p

Angiotensin II Receptor Blocker Ameliorates Stress-Induced Adipose Tissue Inflammation and Insulin Resistance

<div><p>A strong causal link exists between psychological stress and insulin resistance as well with hypertension. Meanwhile, stress-related responses play critical roles in glucose metabolism in hypertensive patients. As clinical trials suggest that angiotensin-receptor blocker delays the onset of diabetes in hypertensive patients, we investigated the effects of irbesartan on stress-induced adipose tissue inflammation and insulin resistance. C57BL/6J mice were subjected to 2-week intermittent restraint stress and orally treated with vehicle, 3 and 10 mg/kg/day irbesartan. The plasma concentrations of lipid and proinflammatory cytokines [Monocyte Chemoattractant Protein-1 (MCP-1), tumor necrosis factor-α, and interleukin-6] were assessed with enzyme-linked immunosorbent assay. Monocyte/macrophage accumulation in inguinal white adipose tissue (WAT) was observed with CD11b-positive cell counts and mRNA expressions of CD68 and F4/80 using immunohistochemistry and RT-PCR methods respectively. The mRNA levels of angiotensinogen, proinflammatory cytokines shown above, and adiponectin in WAT were also assessed with RT-PCR method. Glucose metabolism was assessed by glucose tolerance tests (GTTs) and insulin tolerance tests, and mRNA expression of insulin receptor substrate-1 (IRS-1) and glucose transporter 4 (GLUT4) in WAT. Restraint stress increased monocyte accumulation, plasma free fatty acids, expression of angiotensinogen and proinflammatory cytokines including MCP-1, and reduced adiponectin. Irbesartan reduced stress-induced monocyte accumulation in WAT in a dose dependent manner. Irbesartan treatment also suppressed induction of adipose angiotensinogen and proinflammatory cytokines in WAT and blood, and reversed changes in adiponectin expression. Notably, irbesartan suppressed stress-induced reduction in adipose tissue weight and free fatty acid release, and improved insulin tolerance with restoration of IRS-1 and GLUT4 mRNA expressions in WAT. The results indicate that irbesartan improves stress-induced adipose tissue inflammation and insulin resistance. Our results suggests that irbesartan treatment exerts additive benefits for glucose metabolism in hypertensive patients with mental stress.</p></div

Accumulation of monocytes in inguinal adipose of stressed mice.

<p>Stressed mice were individually subjected to 2 h/day of immobilization stress for two weeks. Animals received oral vehicle, 3, or 10 mg/kg/day of irbesartan during the same period. Inguinal adipose tissues from stressed and control (non-stressed) mice were analyzed by H&E staining (A), CD11b immunostaining (B and C), and quantitative RT-PCR for CD68 and F4/80 (D and E). <b>A:</b> Accumulation of mononuclear cells in inguinal adipose tissues following the 2-week restraint stress. Top panel, ×40 magnification, bar = 250 µm. Inset, ×200 magnification, bar = 50 µm. <b>B:</b> Increased accumulation of CD11b-positive cells (monocytes) in adipose tissue of stressed mice (×200 magnification, bar = 50 µm). <b>C:</b> Quantitative analysis of CD11b-positive cells relative to total nuclear number. Data are mean±SD. n = 10 for all the groups.*<i>P<</i>0.001, compared with the vehicle-treated control mice, **<i>P<</i>0.001, compared with the vehicle-treated and stressed mice. <b>D</b> and <b>E</b>: Quantitative analysis of F4/80 (D) and CD68 (E) expression levels in adipose tissue. Data are mean±SD. n = 10 for all the groups. Values are expressed relative to the vehicle-treated control mice. (<b>D</b>) *<i>P<</i>0.001, compared with the vehicle-treated control mice, **<i>P<</i>0.001, compared with the vehicle-treated and stressed mice, respectively. (<b>E</b>) *<i>P<</i>0.001, compared with the vehicle-treated control mice, ^†<i>P<</i>0.012, compared with vehicle-treated and stressed mice, ^#<i>P<</i>0.02, compared with the stressed mice treated with a lower dose of irbesartan (3 mg/kg/day), respectively.</p

Irbesartan rescued stress-induced decline in insulin sensitivity.

<p><b>A:</b> Glucose tolerance was comparable between the stressed mice treated with vehicle and irbesartan (10 mg/kg/day) after stress. Insulin tolerance showed significant recovery in the irbesartan-treated and stressed mice (lower panel). Data are mean ± SD of 10 mice per group. *<i>P<</i>0.05, and **<i>P<</i>0.02, compared with the vehicle-treated and stressed mice. <b>B:</b> Quantitative analysis of IRS-1 and GLUT4 expression in inguinal adipose tissue and skeletal muscle (adductor muscle) of the stressed mice treated with vehicle or irbesartan (10 mg/kg/day). Data are mean ± SD of 10 mice per group. *<i>P<</i>0.05, compared with the vehicle-treated and stressed mice.</p

Irbesartan reduced the expression of stress-induced proinflammatory adipokines and restored adiponectin expression in adipose tissue.

<p>Inguinal adipose tissues from control mice treated with vehicle or irbesartan (10 mg/kg/day), and stressed mice treated with vehicle or irbesartan (3 or 10 mg/kg/day) were analyzed by quantitative RT-PCR for angiotensinogen (<b>A</b>), MCP-1 (<b>B</b>), TNF-α (<b>C</b>), IL-6 (<b>D</b>), and adiponectin (<b>E</b>). Values are expressed relative to the vehicle-treated control mice. Plasma levels of MCP-1, TNF-α, and IL-6 from these groups were also measured. Data are mean ± SD of 10 mice for RT-PCR, 6 mice for ELISA per group. (<b>A</b>) *<i>P<</i>0.001, compared with the vehicle-treated control mice, **<i>P<</i>0.046, compared with the vehicle-treated and stressed mice, ^†<i>P<</i>0.042, compared with the stressed mice treated with a lower dose of irbesartan (3 mg/kg/day), respectively. (<b>B</b>) *<i>P<</i>0.001, compared with the vehicle-treated control mice, **<i>P<</i>0.001, compared with the vehicle-treated and stressed mice, ^†<i>P<</i>0.003, compared with the vehicle-treated and stressed mice, respectively. (<b>C</b>) *<i>P<</i>0.001, compared with the vehicle-treated control mice, **<i>P<</i>0.001, compared with the vehicle-treated and stressed mice, ^†<i>P<</i>0.004, compared with the stressed mice treated with a lower dose of irbesartan (3 mg/kg/day), ^††<i>P<</i>0.05, compared with the vehicle-treated and stressed mice, respectively. (<b>D</b>) *<i>P<</i>0.001, compared with the vehicle-treated control mice, **<i>P<</i>0.003, compared with the vehicle-treated and stressed mice, ^†<i>P<</i>0.004, compared with stressed mice treated with a lower dose of irbesartan (3 mg/kg/day), ^††<i>P<</i>0.002, compared with the vehicle-treated control mice, ^#<i>P<</i>0.02, compared with the vehicle-treated and stressed mice, respectively. (<b>E</b>) *<i>P<</i>0.001, compared with the vehicle-treated control mice, **<i>P<</i>0.05, compared with the vehicle-treated and stressed mice, respectively.</p