Ethidium bromide-stained polyacrylamide gel showing polymerase chain reaction products derived from gene rearrangements in patients with rheumatoid arthritis and T-cell large granular lymphocyte (T-LGL) proliferations

Polyclonal expansion of T-LGLs in patient 10. Lane 1: gene rearrangement–negative, polyclonal smear (tube A); lane 2: gene rearrangement-negative, polyclonal smear (tube B); lane 3: gene rearrangement-negative, polyclonal smear (tube C); lane 4: standard 50 base pairs (bp); lane 5: gene rearrangement-negative, polyclonal smear (tube A); lane 6: gene rearrangement-negative, polyclonal smear (tube B); and lane 7: gene rearrangement-negative, polyclonal smear. Monoclonal expansion in polyclonal background in patient 7. Lane 1: gene rearrangement: monoclonal product 180 bp (i) in tube A; lane 2: gene rearrangement: monoclonal product 210 bp (ii) in polyclonal background (tube B); lane 3: gene rearrangement: monoclonal product 160 bp (iii); lane 4: (tube A) gene rearrangement-negative, polyclonal smear; lane 5: standard 50 bp; lane 6: (tube B) gene rearrangement-negative, polyclonal smear; and lane 7: (tube C) gene rearrangement-negative, polyclonal smear. Monoclonal gene rearrangements in patient 1 with T-LGL leukemia. Lane 1: gene rearrangement-negative (tube A); lane 2: gene rearrangement-positive, monoclonal product 250 bp (iv) in tube B; lane 3: (tube C): gene rearrangement-negative, polyclonal smear; lane 4: standard 50 bp; lane 5: gene rearrangement-positive, monoclonal product 230 bp (v) in tube A; lane 6: gene rearrangement-positive, monoclonal product 180 bp (vi) in tube B; and lane 7: gene rearrangement-negative, polyclonal smear. TCR, T-cell receptor.<p><b>Copyright information:</b></p><p>Taken from "Characteristics of T-cell large granular lymphocyte proliferations associated with neutropenia and inflammatory arthropathy"</p><p>http://arthritis-research.com/content/10/3/R55</p><p>Arthritis Research & Therapy 2008;10(3):R55-R55.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2483444.</p><p></p

Histopathological features of bone marrow in patients with arthritis and T-cell large granular lymphocyte (T-LGL) lymphocytosis

Patient 1 with rheumatoid arthritis (RA) and T-LGL leukemia. Staining for CD57 demonstrates intrasinusoidal linear arrays and interstitial clusters of T cells (EnVision stain, ×100). Granzyme B highlights cytotoxic granules in these cells (EnVision stain, ×200). Patient 10 with polyclonal T-LGL lymphocytosis. Staining for CD8 shows dispersed T cells (EnVision stain, ×200). Patient 9 with unclassified arthritis, T-LGL leukemia, and and gene rearrangements. CD3 staining shows interstitial and nodular infiltration of T cells (EnVision stain, ×100). Patient 9. The lymphoid nodule contains few CD20B cells (EnVision stain, ×200). Patient 7 with RA and T-LGL leukemia. A decreased count of granulocytic precursors (myeloperoxydase) is shown (EnVision stain, ×200). IGKV, immunoglobulin kappa variable; IGLV, immunoglobulin lambda variable<p><b>Copyright information:</b></p><p>Taken from "Characteristics of T-cell large granular lymphocyte proliferations associated with neutropenia and inflammatory arthropathy"</p><p>http://arthritis-research.com/content/10/3/R55</p><p>Arthritis Research & Therapy 2008;10(3):R55-R55.</p><p>Published online 12 May 2008</p><p>PMCID:PMC2483444.</p><p></p

Prognostic Impact of Copy Number Alterations’ Profile and AID/RAG Signatures in Acute Lymphoblastic Leukemia (ALL) with BCR::ABL and without Recurrent Genetic Aberrations (NEG ALL) Treated with Intensive Chemotherapy

Adult acute lymphoblastic leukemia (ALL) is associated with poor outcomes. ALL is initiated by primary aberrations, but secondary genetic lesions are necessary for overt ALL. In this study, we reassessed the value of primary and secondary aberrations in intensively treated ALL patients in relation to mutator enzyme expression. RT-PCR, genomic PCR, and sequencing were applied to evaluate primary aberrations, while qPCR was used to measure the expression of RAG and AID mutator enzymes in 166 adult ALL patients. Secondary copy number alterations (CNA) were studied in 94 cases by MLPA assay. Primary aberrations alone stratified 30% of the patients (27% high-risk, 3% low-risk cases). The remaining 70% intermediate-risk patients included BCR::ABL1pos subgroup and ALL lacking identified genetic markers (NEG ALL). We identified three CNA profiles: high-risk bad-CNA (CNAhigh/IKZF1pos), low-risk good-CNA (all other CNAs), and intermediate-risk CNAneg. Furthermore, based on RAG/AID expression, we report possible mechanisms underlying the CNA profiles associated with poor outcome: AID stratified outcome in CNAneg, which accompanied most likely a particular profile of single nucleotide variations, while RAG in CNApos increased the odds for CNAhigh/IKZF1pos development. Finally, we integrated primary genetic aberrations with CNA to propose a revised risk stratification code, which allowed us to stratify 75% of BCR::ABL1pos and NEG patients