In vivo expression of the HBZ gene of HTLV-1 correlates with proviral load, inflammatory markers and disease severity in HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP)

<p>Abstract</p> <p>Background</p> <p>Recently, human T-cell leukemia virus type 1 (HTLV-1) basic leucine zipper factor (HBZ), encoded from a minus strand mRNA was discovered and was suggested to play an important role in adult T cell leukemia (ATL) development. However, there have been no reports on the role of HBZ in patients with HTLV-1 associated inflammatory diseases.</p> <p>Results</p> <p>We quantified the HBZ and tax mRNA expression levels in peripheral blood from 56 HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients, 10 ATL patients, 38 healthy asymptomatic carriers (HCs) and 20 normal uninfected controls, as well as human leukemic T-cell lines and HTLV-1-infected T-cell lines, and the data were correlated with clinical parameters. The spliced HBZ gene was transcribed in all HTLV-1-infected individuals examined, whereas tax mRNA was not transcribed in significant numbers of subjects in the same groups. Although the amount of HBZ mRNA expression was highest in ATL, medium in HAM/TSP, and lowest in HCs, with statistical significance, neither tax nor the HBZ mRNA expression per HTLV-1-infected cell differed significantly between each clinical group. The HTLV-1 HBZ, but not tax mRNA load, positively correlated with disease severity and with neopterin concentration in the cerebrospinal fluid of HAM/TSP patients. Furthermore, HBZ mRNA expression per HTLV-1-infected cell was decreased after successful immunomodulatory treatment for HAM/TSP.</p> <p>Conclusion</p> <p>These findings suggest that <it>in vivo </it>expression of HBZ plays a role in HAM/TSP pathogenesis.</p

Predictive impact of soluble interleukin‐2 receptor and number of extranodal sites for identification of patients at very high risk of CNS relapse in diffuse large B‐cell lymphoma

Abstract There remains an unmet clinical need to identify which patients with diffuse large B‐cell lymphoma (DLBCL) would benefit from central nervous system (CNS) prophylaxis, due to the low positive predictive value (PPV; 10%–15%) of the currently available predictive models. To stratify patients at high risk of developing CNS relapse, we retrospectively analyzed 182 patients with DLBCL initially treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R‐CHOP), or a R‐CHOP‐like regimen. Among them, 17 patients relapsed with CNS involvement, and the 2‐year rate of CNS relapse was 7.9%. Upon carrying out multivariate analysis, ≥3 extranodal sites and elevated soluble interleukin‐2 receptor (sIL‐2R) levels at diagnosis were identified as independent risk factors for CNS relapse. The 2‐year and 3.5‐year rates of CNS relapse were 57.1% and 78.6%, respectively, in patients with both elevated sIL‐2R and ≥3 extranodal sites. Furthermore, combined use of these risk factors of both elevated sIL‐2R and ≥3 extranodal sites resulted in a high PPV (71.4%), negative predictive value (93.1%), and overall accuracy (92.3%) for undergoing CNS relapse. In conclusion, we propose a simple and valuable tool to predict patients with DLBCL at very high risk of CNS relapse

Waldenstrom's macroglobulinemia-like B cell lymphoma with MYD88 L265P mutation and t(14;18)(q32;q21) involving IGH-MALT1

A 65-year-old woman was referred to the hospital for further investigation of weight loss, hyperproteinemia, and anemia. Serum immunofixation electrophoresis revealed IgM-κ M protein. Bone marrow examination revealed an increase in the number of B -cells with immunoglobulin kappa light-chain restriction. Although the MYD88 L265P mutation was identified in bone marrow mononuclear cells, which suggested the diagnosis of Waldenstrom's macroglobulinemia (WM), a fusion signal of IgH-MALT1, which is commonly observed in extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) lymphoma, was also identified. Here, we describe a rare case of low-grade B-cell lymphoma with MYD88 L265P mutations accompanying IgH-MALT1

Top 15 clones of each cell lineage in HAM/TSP#2.

<p>Top 15 clones of each cell lineage in HAM/TSP#2.</p

HTLV-1 infection in multiple lineage hematopoietic cells.

<p>Bone marrow cells from two STLV-1 infected JMs were cultured for 24 hours after removal of CD8⁺ T cells. (A, B) Tax expression in bone marrow cells of two STLV-1 infected JMs. Mononuclear cells from bone marrow were stained by various antibodies. Tax expression was shown in association with CD3, CD4, CD8, CD34 and CD33 expression.</p

Top 15 clones of each cell lineage in HAM/TSP#3.

<p>Top 15 clones of each cell lineage in HAM/TSP#3.</p

HTLV-1 infection in neutrophils and monocytes.

<p>(A) Tax expression is detected by confocal immunofluorescence microscopy in MT4 and neutrophils from HAM/TSP patients using anti-Tax antibody and secondary Alexa Fluor 488 antibody. Myeloperoxidase expression is also detected in neutrophils using anti-MPO antibody and secondary Alexa Fluor 568 antibody. Jurkat was used as a negative control. MT4, HTLV-1 infected cell line; Jurkat cells, HTLV-1 negative human T-cell line. DAPI (blue) was used for nuclei staining. (B) JET WT35 cells were co-cultured with HPB-ATL-2 cells in the presence and absence of azidothymidine (AZT) or raltegravir (RAL), or HTLV-1 uninfected T cell line, CCRF-CEM. The number of tdTomato-positive cells is presented. (C) Dendritic cells were induced from monocytes from HAM/TSP patients, and then differentiated DCs were co-cultured with JET WT35. After 48 hours, newly infected JET WT35 was shown to be red. (D) Quantitative data of co-culture between differentiated DCs and JET WT35 cells. The count of tdTomato positive cells and the number of cells used for this experiment are shown.</p

The proportions of clones with the same integration sites.

<p>The proportions of clones with the same integration sites are shown. The same integration sites are identified in 5 (red), 4 (yellow), 3 (light blue), and 2 (blue) different lineage cells. For HAM/TSP1, the proportion of clones with the same integration sites between PBMCs and neutrophils are shown in orange.</p