Galectin-1 (Gal-1) has been associated with adverse prognosis in several cancers including lymphoma entities with CD30 expression. However, Gal-1 expression has not been systematically assessed in peripheral T-cell lymphomas (PTCL). Specimens from 169 nodal PTCL were assessed for intratumoural Gal-1 expression by immunohistochemistry. Overall survival (OS) in groups exhibiting high and low Gal-1 expression was compared in the cohort and in a subset analysis of CD30-positive PTCL only. Gal-1 expression was also correlated with biomarkers of the tumour microenvironment. No significant difference in OS based on Gal-1 expression was observed in the entire PTCL cohort. However, in the CD30-positive cohort, patients with high Gal-1 levels had significantly poorer outcome (5 years OS 10%, 95% confidence interval CI, 1-36) than their low Gal-1 counterparts (5 years OS 48%, 95% CI, 30-64, P = .021). In univariate analyses age 60 or younger, non-elevated lactate dehydrogenase (LDH), and performance score less than 2 correlated with superior survival but high Gal-1 expression significantly predicted adverse outcome at both univariate (HR 2.5, 95% CI, 1.1-5.7, P = .026) and multivariate levels (HR 3.2, 95% CI, 1.2-8.5, P = .017). Tumours with high Gal-1 had few cytotoxic T cells in the tumour microenvironment. High intratumoural Gal-1 expression before therapeutic intervention correlates with adverse outcome in nodal CD30(+), ALK(-) PTCL patients.

Peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) constitutes a heterogeneous category of lymphomas, which do not fit into any of the specifically defined T-cell lymphoma entities. Both the pathogenesis and tumor biology in PTCL-NOS are poorly understood. Protein expression in pretherapeutic PTCL-NOS tumors was analyzed by proteomics. Differentially expressed proteins were compared in 3 distinct scenarios: (A) PTCL-NOS tumor tissue (n = 18) vs benign lymphoid tissue (n = 8), (B) clusters defined by principal component analysis (PCA), and (C) tumors from patients with chemosensitive vs refractory PTCL-NOS. Selected differentially expressed proteins identified by proteomics were correlated with clinico-pathological features and outcome in a larger cohort of patients with PTCL-NOS (n = 87) by immunohistochemistry (IHC). Most proteins with altered expression were identified comparing PTCL-NOS vs benign lymphoid tissue. PCA of the protein profile defined 3 distinct clusters. All benign samples clustered together, whereas PTCL-NOS tumors separated into 2 clusters with different patient overall survival rates (P = .001). Differentially expressed proteins reflected large biological diversity among PTCL-NOS, particularly associated with alterations of "immunological" pathways. The 2 PTCL-NOS subclusters defined by PCA showed disturbance of "stress-related" and "protein metabolic" pathways. α-Enolase 1 (ENO1) was found differentially expressed in all 3 analyses, and high intratumoral ENO1 expression evaluated by IHC correlated with poor outcome (hazard ratio, 2.09; 95% confidence interval, 1.17-3.73; P = .013). High expression of triosephosphate isomerase (TPI1) also showed a tendency to correlate with poor survival (P = .057). In conclusion, proteomic profiling of PTCL-NOS provided evidence of markedly altered protein expression and identified ENO1 as a novel potential prognostic marker.

At HIV-1 infection, the binding of the viral envelope proteins to CD4+ is essential for viral transmission, and this process is facilitated by interaction with the highly conserved host lectin, galectin-1 (Gal-1) [1–3]. Within the tumor microenvironment, Gal-1 is expressed by both tumor and stromal cells where it promotes tumor immune escape and favors hypoxia-driven angiogenesis [4–6]. In sporadically occurring Hodgkin lymphoma, high Gal-1 expression at diagnosis is associated with poorer treatment response [7], and high soluble Gal-1 (sGal-1) correlates with adverse disease characteristics [8]. Previous studies have shown that targeted inhibition of Gal-1 prevents tumor-induced immunosuppression [9,10] and inhibits tumor growth and metastasis in various tumor models [6,11–13]. Recently, we published a proteomic profiling study of pretreatment serum samples from HIV-infected patients, identifying several differentially expressed proteins associated with lymphoma development [14]. In this cohort, we have now evaluated serum levels of sGal-1 and correlated this with clinical parameters, including lymphoma development. In addition, we have investigated the intratumoral expression and prognostic value of Gal-1 in HIV-associated lymphomas, and, for comparison, sGal-1 serum levels in 30 healthy blood donors [15] Circulating sGal-1 levels were measured using a time-resolved immunofluorometric assay and immunohistochemistry and the evaluation of tumoral Gal-1 expression were performed as described previously [7,14,15]. Pretreatment sGal-1 serum levels were assessed in 19 HIV-positive individuals at time of HIV diagnosis. There were no sex-related differences (P = 0.450) and sGal-1 levels neither correlate with peripheral CD4+ cell count nor with viral load at HIV diagnosis (ρ = −0.491 P = 0.852 and ρ = −0.009 P = 0.974, respectively). HIV-infected individuals had significantly lower levels of sGal-1 compared with healthy controls (43.6 vs. 84.9 ng/ml; P < 0.001; Fig. 1a). Within the entire study cohort (healthy controls and HIV-infected individuals), those patients who would later develop lymphoma also had significantly lower levels of sGal-1 at time of HIV-diagnosis (Fig. 1b; P = 0.016). There was no significant difference in sGal-1 within the HIV cohort (Fig. 1c, P = 0.130).Fig. 1: Serum galectin-1 (Gal-1) levels in HIV-infected individuals and controls. The line indicates the median and the box indicates the 25th and 75th percentiles. Whiskers are upper and lower adjacent values.(a) HIV-infected individuals had significantly lower levels of soluble Gal-1 at the time of HIV diagnosis compared with healthy controls (P < 0.001). (b) Soluble Gal-1 at the time of HIV diagnosis in HIV-infected individuals with subsequent lymphoma compared with the remaining cohort (healthy controls and HIV-infected individuals without lymphoma) (P < 0.016). (c) Differences in Gal-1 serum levels in HIV-infected individuals who did or did not develop lymphoma (P < 0.130).A cut-off value of 2.4 ng/ml generated by receiver operating curve (ROC) analysis separated HIV-infected individuals who later developed lymphoma from the remaining cohort of HIV patients and controls with a specificity of 82% and a sensitivity of 100%. Based on this cut-off value, 13 (31%) HIV-infected patients were allocated to the low sGal-1 group, including all future lymphoma patients (N = 5). Tumoral Gal-1 expression correlated positively with a proinflammatory signature of the microenvironment, including the macrophage marker CD68, the cytotoxic markers CD8 and granzyme B, as well as the activation marker CD30 [CD68 (ρ = 0.740; P < 0.001), CD8 (ρ = 0.379; P = 0.027), granzyme B (ρ = 0.579; P < 0.001) and CD30 (ρ = 0.467; P = 0.006)]. Clinical features of the cohort included in the tissue microarray have been described previously [14]. Gal-1 was widely expressed in all lymphoma subtypes. Based on a ROC generated cut-off value for high vs. low intratumoral Gal-1 expression, 59% (N = 10) of diffuse large B-cell lymphoma (DLBCL) patients had a high level of intratumoral Gal-1 expression (>24.8% positive cells). In the total lymphoma cohort (all diagnoses), two-thirds of the patients (N = 22; 65%) were high expressers. This latter group more often had nodal disease and B-symptoms (P = 0.006). Gal-1 did not correlate with tumoral Epstein–Barr virus (EBV) status, EBV latency type, international prognostic index (IPI), clinical stage, or cell of origin. In HIV-associated DLBCL, patients with higher levels of intratumoral Gal-1 expression had a significantly better outcome with a 5-year overall survival of 70.0% (95% confidence interval 32.9–89.2%) vs. 14.3% (95% confidence interval 0.7–46.5%). In a multivariate analysis, adjusting for IPI and rituximab treatment, both Gal-1 expression (P = 0.021) and IPI (P = 0.049) retained an independent prognostic value. HIV infection has a profound influence on the host immune system including altered cytokine and protein expression years prior to lymphoma diagnosis [14,16–18]. Gal-1 is secreted by most immune cells [19] and the significantly lower levels of sGal-1 in newly diagnosed HIV-infected individuals (compared with healthy controls), as found in our study, may reflect the dramatically altered immune constitution of these patients. This may lead to a proinflammatory although nonefficient T-cell response, ultimately leading to lymphoma development. We found a relatively high intratumoral expression of Gal-1 in our cohort of HIV-associated DLBCL, as compared with the immunocompetent setting [20]. This may partly reflect different evaluation techniques, but inherent disparities in lymphoma microenvironment may also be involved [21,22]. Gal-1 is largely produced by macrophages [23]. The correlation between high Gal-1 expression and improved outcome in HIV-associated DLBCL may therefore be explained by a higher level of macrophages because they have been shown to improve the efficacy of antibody-driven immunotherapy [24]. In conclusion, the results of our study indicate that Gal-1 is significantly associated with risk of lymphoma in HIV-infected individuals and may represent an attractive future target for the management of HIV-associated lymphoma. Acknowledgements The authors wish to thank Erik Hagen Nielsen, Vibeke Ellerup Jensen, and Kristina Lystlund Lauridsen for expert technical assistance, and Betina S. Sørensen for facilitating access to healthy donors samples. Conceived and designed the study: M.Ø.V., M.L., C.S.L., F.d’A. Provided study material: G.A.R., I.P., G.P., C.S.L., M.B.M., K.B., S.H.D. Performed the experiments: R.H., M.L. Analyzed data: M.Ø.V., M.L., B.H., P.W.D. Wrote the paper: M.Ø.V. and M.L. Final editing and approval of the manuscript: all authors. The work was supported by unrestricted grants from Dagmar Marshalls Foundation, Manufacturer Einer Willumsen's Memorial Foundation, The Harboe Foundation, The Krista and Viggo Petersens Foundation, Fonden til Lægevidenskabens fremme, Director Emil C. Hertz and his wife Inger Hertz Foundation, The Foundation of 17 December 1981, Architect Holger Hjortenberg and wife Dagmar Hjortenberg's Foundation; Frits, Georg, and Marie Cecilie Glud's Foundation, Danish Diabetes Academy supported by the Novo Nordisk Foundation, Inger and Max Wørzner's Memorial Foundation, and The MEMBRANES Center at Aarhus University. Conflicts of interest There are no conflicts of interest.

Galectin-1 (Gal-1) has been associated with prognosis in several cancers including lymphomas. CD30-dependent cell signaling mediated by Gal-1 has been proposed in CD30-positive lymphomas and cell lines. Still, Gal-1 expression has not been systematically assessed as a possible prognosticator in nodal peripheral T-cell lymphomas (PTCL).

To the editor: Peripheral T-cell lymphomas (PTCLs) represent a group of rare hematological cancers of mature T-cell or natural killer cell origin accounting for 10% to 15% of all lymphomas.[1][1] Although many patients have poor outcomes, some achieve long-term survival.[2][2],[3][3] Thus,

investigated. We hypothesized that rituximab‐based therapy would improve survival in B‐cell–mediated AID‐associated lymphoma, given its beneficial clinical effect in AID. We aimed to examine the association between pre‐existing AIDs and B‐NHL and the possible influence of AIDs on NHL outcome. Methods: In this hospital‐based case‐control study in Hadassah– Hebrew University Medical Center, we recruited 435 newly‐diagnosed adult (>18 years) CD20+ B‐NHL patients diagnosed 2009 to 2014 and 414 controls frequency‐matched by age and sex to cases. The study is based on questionnaires in Hebrew, English, and Russian that included sociodemographic variables, medical information including a history of AIDs and medications; pathology confirmation; and chart review. We examined the association between NHL and AIDs in general, B‐ and T‐cell–mediated AIDs and autoimmune thyroid diseases, using logistic regression, reporting odds ratios (OR), and 95% confidence intervals (CI). In the second part of the study, we compared overall (OS) and relapse‐free survival (RFS) in B‐NHL patients with and without AID. We constructed Kaplan‐Meier curves for univariate analysis, and multivariable Cox regression models adjusting for important patient and disease characteristics such as Ki67% staining, international prognostic index score (IPI), and histological subgroup. Results: B‐NHL risk was associated with the presence of AIDs (OR = 1.98; 95% CI, 1.01‐3.9) especially those mediated by B‐cell activation (OR = 5.97; 95% CI, 2.3‐15.6). The strongest association was observed for marginal zone lymphoma (OR = 13.2, 95% CI, 4.02‐43.6). Time to relapse for all B‐NHL patients with AIDs was significantly shorter (mean of 49.21 months [±3.22]) than for patients without AID (mean of 59.74 months [±1.62]), hazard ratio (HR) = 1.7 (95% CI, 1.03‐2.79), after adjusting for IPI, Ki67% staining, and histological subgroup. Specifically in DLBCL, in which >99% received rituximab‐based therapy, both RFS and OS were adversely affected by the presence of B‐cell mediated AIDs with HR = 7.84 (95% CI, 2.86‐21.5) and 3.99 (95% CI, 1.24‐12.6), respectively (see figure). Conclusions: Beyond the well‐known association between AIDs and B‐NHL (particularly AIDs mediated by B‐cell activation), we found in addition that AID is an adverse prognostic factor in B‐cell lymphoma. AID‐associated B‐NHL patients have poorer outcomes. Specifically, B‐cell mediated AID results in inferior RFS and OS in DLBCL, suggesting that rituximab‐based therapy does not provide adequate coverage for the subgroup of patients. Further exploration of molecular subtypes and mechanisms of resistance of B‐NHL associated with AID is warranted.

Post-transplant lymphoproliferative disorder (PTLD) incidence is difficult to determine, mainly because both early and other lesions may go unrecognized and unregistered. Few studies have included systematic pathology review to maximize case identification and decide more accurately PTLD frequency after long-term post-transplantation follow-up. A retrospective population-based cohort study including all kidney transplant recipients at two Danish centres (1990-2011; population covered 3.1 million; 2175 transplantations in 1906 patients). Pathology reports were reviewed for all patient biopsies to identify possible PTLDs. Candidate PTLDs underwent histopathological review and classification. Seventy PTLD cases were identified in 2175 transplantations (3.2%). The incidence rate (IR) after first transplantation was 5.4 cases per 1000 patient-years (95% CI: 4.0-7.3). Most PTLDs were monomorphic (58.5%), or early lesions (21.5%). Excluding early lesions and patients <18 years, IR was 3.7 (95% CI: 2.9-5.5). Ten patients with PTLD were retransplanted, 2 developing further PTLDs. Post-transplant patient survival was inferior in patients with PTLD, while death-censored graft survival was not. Using registry data together with extensive pathological review and long follow-up, a rather high incidence of PTLD was found.

Objective: HIV-infected individuals have an increased risk of developing lymphoma. We sought to identify markers predictive of lymphoma development by comparing protein expression patterns in serum obtained at the time of HIV diagnosis from patients who later developed malignant lymphoma or benign lymphadenopathy, with samples from patients with no subsequent history of neoplasia.Design: All patients were identified retrospectively from the Danish HIV cohort.Methods: Serum samples (N = 21), obtained at time of HIV diagnosis, were subjected to high-resolution two-dimensional gel electrophoresis. Differentially expressed proteins were identified by liquid chromatography-tandem mass spectrometry. A tissue microarray, containing diagnostic HIV-lymphoma tissue samples (N = 40), was used to investigate immunohistochemical expression of markers in tumoural lesions.Results: Fourteen differentially expressed protein spots were detected. Using principal components analysis, spots containing immunoglobulin J chain, apolipoprotein A-I, procollagen C-endopeptidase enhancer-1 and complement C4-A were associated with lymphoma development (P < 0.0001). Serum amyloid A-2 was increased almost 10-fold in patients with subsequent lymphoma compared with patients without subsequent lymphoma. In the tissue microarray, amyloid A was widely expressed, and high expression showed a tendency towards inferior outcome (log-rank 0.073).Conclusion: We identified several differentially expressed protein spots present already at the time of HIV diagnosis. Analysis of biological differences correlating to lymphoma development at this early stage of a possible malignant transformation may lead to the identification of predictive markers. Further investigation of the potential clinical application of differentially expressed proteins as risk stratification markers for monitoring HIV-positive individuals is warranted. Copyright (C) 2016 Wolters Kluwer Health, Inc. All rights reserved.

Introduction. HIV infected individuals have an increased risk of developing lymphoma even in the era of combined antiretroviral therapy. Galectin-1 (Gal-1) is known to promote various immunomodulatory functions, including Treg expansion (Dalotto-Moreno et al, Cancer Res 2013), promotion of tolerogenic dendritic cells (Ilarregui et al, Nat Immunol 2009) and apoptosis of fully-differentiated effector T-cells (Toscano et al, Nat Immunol 2007). In the context of cancer, Gal-1 is expressed on both tumor cells and cells in the tumor microenvironment, and is usually associated with immune privilege, tumor escape and hypoxia-driven angiogenesis (Juszczynski et al, Proc Natl Acad Sci 2007; Cedeno-Laurent et al, Blood 2012). Previously, high intratumoral Gal-1 levels have been suggested as an unfavorable outcome predictor in patients with classical Hodgkin lymphoma (cHL) (Kamper et al, Blood 2011). Furthermore, several in vitro studies revealed the benefit of Gal-1 inhibition with regards to overcoming treatment resistance e.g., after anti-VEGF and anti-CD20 therapy (Croci et al, Cell 2014; Lykken et al, Blood 2016). Thus, Gal-1 inhibition may prospectively be an important tool in lymphoma treatment. In this study, we have investigated the Gal-1 expression in pre-therapeutic tumoral tissue samples from patients with HIV-associated lymphoma and its correlation to clinicopathological features at lymphoma diagnosis.

The tumour microenvironment in classical Hodgkin's lymphoma (cHL) is characterised by a minor population of neoplastic Hodgkin and Reed-Sternberg cells within a heterogeneous background of non-neoplastic bystanders cells, including mast cells. The number of infiltrating mast cells in cHL has been reported to correlate with poor prognosis. We used immunohistochemistry to assess the degree of tumour-infiltrating mast cells in cHL tissue microarrays and correlated this with clinico-pathological features and prognosis in a cohort of homogeneously treated patients with Hodgkin's disease. A high degree of tumour mast cells was associated with nodular sclerosis (NS) subtype histology (P = 0.0002). Moreover, the number of mast cells was inversely correlated with the numbers of CD68+ and CD163+ macrophages (P = 0.0001 and P = 0.003, respectively) and with the number of granzyme+ cytotoxic cells (P = 0.004). The degree of mast cell infiltration was not a prognostic factor in cHL of nodular sclerosis subtype. In contrast, in mixed cellularity cHL a high number of intratumoral mast cells correlated with significantly poorer outcome both in terms of overall (P = 0.03) and event-free survival (P = 0.01). Further studies are warranted into the biological mechanisms underlying this adverse outcome and their possible therapeutic implications.

In W estern countries, the age distribution of H odgkin lymphoma ( HL ) follows a characteristic bimodal curve showing an early and a late peak at approximately 35 and 70 yr, respectively. Furthermore, the presence of latent E pstein‐ B arr virus ( EBV ) genome in the H odgkin R eed– S ternberg cells, the tumour cell population of classical HL (c HL ), has been found to have adverse prognostic impact in elderly, but not in younger c HL patients. We have characterised the protein expression in tumour tissue samples from younger (≤55 yr) and elderly (>55 yr) c HL patients and correlated the findings with EBV status. Differentially expressed proteins according to patients’ age as well as tumoural EBV status belonged to different biological functional domains, such as apoptosis, cytoskeletal organisation, response to oxygen levels and regulation of catabolic/metabolic processes. The differential expression of selected proteins, cytosolic aminopeptidase, heterogeneous nuclear ribonucleoprotein K , serotransferrin and alpha‐1‐antitrypsin was further validated by W estern blot analysis. Discovery‐based proteomics characterising biological features distinctive for subsets of c HL patients may be useful for the identification of novel biomarkers with potential therapeutic relevance. An evaluation of the prognostic impact of protein expression pattern in general and individually expressed proteins in particular is warranted.

The majority of untransformed follicular lymphoma (FL) follow an indolent clinical course and have a median overall survival that, in several series, exceeds one decade.1, 2, 3 Histological transformation (HT), usually to diffuse large B-cell lymphoma (DLBCL), occurs in ~30% of all patients with grade I/II FL.4, 5, 6 HT is usually associated with a rapidly progressive clinical course, treatment resistance and poor survival. Although HT is a well-described clinico-pathological event, the molecular mechanisms behind it are still largely unknown, particularly regarding changes in global protein expression. Moreover, no unequivocal prognostic tools have been identified to effectively predict the patients at risk of HT.

Post-transplant lymphoproliferative disorders (PTLDs) are potentially fatal, often Epstein-Barr virus (EBV)-driven neoplasias developing in immunocompromised hosts. Initial treatment usually consists of a reduction in immunosuppressive therapy and/or rituximab with or without chemotherapy. However, patients who relapse do poorly, and new treatment options are warranted. With the introduction of the immunoconjugate brentuximab vedotin, the CD30 antigen has become an effectively targetable molecule. Therefore, we investigated the frequency and level of CD30 expression in PTLDs. We identified 108 patients with PTLDs diagnosed during 1994-2011, of whom 62 had adequate paraffin-embedded tissue for tissue microarray construction. Immunohistochemical expression of CD30 was consistently detected in all types of PTLD (overall 85.25%), including the monomorphic subtypes, and was correlated with a more favorable outcome. For diffuse large B-cell lymphoma (DLBCL)-type PTLD this was regardless of EBV status, and remained significant in multivariate analysis. Cell-of-origin had no independent prognostic value in our series of DLBCL PTLD.

Background. Posttransplant lymphoproliferative disorder (PTLD) is a feared complication to organ transplantation, associated with substantial morbidity and inferior survival. Risk factors for PTLD include T cell-depleting induction therapy and primary infection or reactivation of Epstein-Barr virus. Possible associations between certain HLA types and the risk of developing PTLD have been reported by other investigators; however, results are conflicting. Methods. We conducted a retrospective, population-based study on 4295 Danish solid organ transplant patients from the Scandiatransplant database. Having identified 93 PTLD patients in the cohort, we investigated the association of HLA types with PTLD, Epstein-Barr virus status and time to PTLD onset. The outcomes survival and PTLD were evaluated using Cox regression; mismatching, and the PTLD-specific mortality were evaluated in a competing risk analysis. Results. Risk of PTLD was associated with male sex (odds ratio, 1.70; 95% confidence interval, 1.07-2.71), and, in women, HLA-DR13 conferred an increased risk (odds ratio, 3.22; 95% confidence interval, 1.41-7.31). In multivariate analysis, HLA-B45 and HLA-DR13 remained independent predictive factors of PTLD. Mismatching in the B locus was associated with a reduced risk of PTLD (P < 0.001). Overall survivalwas poor after a PTLD diagnosis and was significantly worse than that in the remaining transplant cohort (P < 0.001). Conclusions. Our data indicate risk-modifying HLA associations, which can be clinically useful after transplantation in personalized monitoring schemes. Given the strong linkage disequilibrium in the HLA region, the associations must be interpreted carefully. The large size, virtually complete ascertainment of cases and no loss to follow-up remain important strengths of the study.

Introduction: Peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) is a heterogeneous group of mature T-cell lymphomas, probably composed by different biologically related subsets that have not yet been conclusively identified. In the WHO classification, PTCL-NOS accounts for 25-30% of all mature T-/NK-cell malignancies. The clinical outcome is generally poor with a 5-yr overall survival of 30-35% after conventional treatment strategies. The aim of the study was to apply proteomic analysis in PTCL-NOS and to use the protein expression profiles to characterize clinically relevant subsets within this heterogeneous entity by means of unsupervised cluster analysis. Methods: Archival frozen tumor tissue samples from 20 patients diagnosed with PTCL-NOS from 1991 to 2010 were analyzed for protein expression by high-resolution two-dimensional gel electrophoresis. Individual protein spots were visualized with fluorescence staining and the expression profiles were identified. All patients were homogeneously treated with curatively intended anthracycline-containing combination regimens. Clinico-pathological features were obtained from the Danish Lymphoma Registry (LYFO) and from patient records. Hyperplastic tonsils from healthy adults were included as reference tissue (n=8). Principal component analysis and unsupervised hierarchical cluster analysis were performed on the basis of the protein expression profiles. Differentially expressed (two-fold or higher, Mann-Whitney U-test) proteins between the detected clusters were identified by liquid chromatography - tandem mass spectrometry. Results: Unsupervised cluster analysis defined three distinct clusters: one containing all reference samples and two additional ones further subdividing the PTCL-NOS cases in two separate subsets. Patients from these two PTCL-NOS subsets had significantly different responses to treatment and survival (p = 0.001). The differentially expressed proteins were primarily involved in (i) promotion of tumor growth, (ii) regulation of cellular metabolism, and (iii) immune responses. Conclusion : Proteomic analysis identified shared protein expression patterns and potential prognostic markers in subsets of PTCL-NOS. Disclosures No relevant conflicts of interest to declare.

BACKGROUND Transformation of indolent lymphomas (IL) to an aggressive histology (TIL) often results in a rapid clinical course, treatment refractoriness and shortened survival. Although rituximab-containing regimens (R-chemo) have become standard of care in CD20-positive TIL, the role of autologous stem-cell transplantation (ASCT) is still debated. The purpose of this study was to determine whether the outcome of TIL patients improved if they, at transformation, also received ASCT. Furthermore, we investigated the outcome of cases with histologically low- and high-grade components diagnosed either simultaneously or after a period of overt indolent disease. We also analyzed, whether prior rituximab treatment during the indolent course of the disease affected outcome after transformation. PATIENTS AND METHODS Eighty-five patients (≤68 years) with histologically confirmed TIL were included. Five-year overall (OS) and progression-free survival (PFS) were calculated. Selected parameters were tested in a multivariate analysis. All analyses were conducted on three cohorts: (i) whole cohort (all TIL), (ii) patients with co-existing evidence of both indolent and aggressive histology at diagnosis (Composite/discordant TIL) and (iii) patients transformed after prolonged prior indolent disease (sequential TIL). RESULTS Fifty-four patients (64%) received ASCT consolidation and 31 (36%) did not. Within the 'all TIL' cohort, the 5-year OS and PFS for R-chemo + ASCT versus R-chemo alone, were 67% versus 48% (P = 0.11) and 60% versus 30% (P = 0.02), respectively. Furthermore, in 'Composite/discordant TIL' R-chemo + ASCT showed no impact on OS (76% versus 67%; P = 0.66) or PFS (71% versus 62%; P = 0.54). Conversely, R-chemo + ASCT improved the outcome of 'sequential TIL' (OS 62% versus 36%; P = 0.07; PFS 53% versus 6%; P = 0.002), regardless of prior rituximab therapy. The beneficial effect of ASCT was significantly higher in patients who had not received rituximab at IL stage. CONCLUSIONS ASCT improved the outcome in sequential, but not composite/discordant TIL. The beneficial impact of ASCT was greater in patients, who were rituximab-naïve at transformation.

Background Limited data exist on the presence and function of immune-competent cells in chronic tendinopathic tendons, and their potential role in inflammation and tissue healing. Objectives To quantify subtypes of immune-competent cells in biopsies from non-ruptured chronic tendinopathic Achilles tendons and healthy control tendons. Methods Fifty patients with non-ruptured chronic Achilles tendinopathy and 15 healthy subjects were included. At time of inclusion, the Achilles tendons were examined clinically and evaluated with ultrasound with regard to tendon thickness and Doppler flow grade immediately before an ultrasound-guided tendon biopsy was obtained. Tissue samples were evaluated immunohistochemically by quantifying the presence of macrophages (CD68-KP1+), iron+ hemosiderophages (Perls Blue), T-lymphocytes (CD3+), B-lymphocytes (CD20+), natural killer cells (CD56+), mast cells (NaSDCl+), Schwann cells (S100+) and endothelial cells (CD34+) using a stereological technique. Cell counts were expressed as number of positive cell profiles per biopsy area (c/a), except for CD68-KP1 and CD34, which were quantified using a point counting grid to estimate number of grid hits of stained cells which were then calibrated to the total biopsy area (expressed as cell fractional area (%)). Descriptive data are reported as median and range if nothing else specified. Comparisons between groups were made with Mann-Whitney U tests or Fisher9s Exact Tests. The level of significance was set at P <0.05. Results Macrophages, T-lymphocytes, mast cells, and natural killer cells were observed in the majority (range: 52–96%) of biopsies from non-ruptured chronic tendinopathic Achilles tendons. Conclusions This study provides evidence for the presence of immune-competent cells in the majority of biopsies from non-ruptured chronic tendinopathic Achilles tendons. Macrophages and endothelial cells were significantly more numerous in chronic tendinopathic tendons than in healthy tendons. Disclosure of Interest : None declared DOI 10.1136/annrheumdis-2014-eular.1684

Systematic validation of construction and analysis parameters when using tissue microarray (TMA) in rare, morphologically heterogenous entities such as peripheral T-cell lymphoma (PTCL) is not reported. We describe a tissue-saving virtual TMA to predetermine the number of cores needed to represent whole tissue sections (WTS) from the same biopsies, using automated and traditional manual methods for the quantification of immunohistochemical stains. Whole paraffin hematoxylin and eosin- and immunohistochemical (CD2, CD30, and Ki-67)-stained sections from 30 PTCLs were digitalized. A virtual TMA with six 1-mm cores per slide was designed to compare agreements in the immunohistochemical scoring. Using digital image analysis and manual stereological counting, immunohistochemical positivity was quantified. Associations were analyzed using the Bland-Altman and correlation plots. In PTCL, we report that 4 cores are required to represent WTS results (ie, agreement within ±10%). High concordance was demonstrated between digital results obtained with WTS compared with 4-core virtual TMA (correlation coefficients: 0.89-0.98), and in the comparative evaluation of 4-core virtual TMA by digital image analysis versus manual stereology (correlation coefficients: 0.91 to 0.99). Virtual TMAs provide an efficient tool for optimizing and validating TMA construction parameters when planning a study. The method can be applied to the same tissues used in a subsequent formal study, without wasting scarce tissue resources. In PTCL, TMAs constructed with four 1-mm cores are representative of WTS. In parallel tests using TMAs and WTS from PTCLs, there is a high level of agreement comparing automated digital with manual stereological methods for the quantification of immunohistochemical biomarker staining.

Background: PTLD comprises a diverse spectrum of hematological conditions, ranging from early lesions, characterized by reactive-like proliferations, to monomorphic lesions, resembling overt lymphoma. Â In most cases, the lesions are believed to arise as the result of reduced immune surveillance secondary to the use of immunosuppressive drugs post-transplant. This view is supported by the observation that PTLDs, particularly those characterized by early or polymorphic lesions, may regress spontaneously upon reduction of the immunosuppressive treatment. Studies in sporadic lymphomas have identified distinct microenvironmental characteristics, predictive of the clinical behavior, but data is scarce in the immunocompromised setting. Therefore, the aim of this study was to investigate the tumor microenvironment in a population-based cohort of PTLD.

Background: T-cell malignancies originating from regulatory T (Treg) cells are almost exclusively confined to human T-cell leukemia virus 1 (HTLV-1) associated adult T-cell leukemia/lymphoma (ATLL), although sporadic cases of other peripheral T-cell lymphomas (PTCLs) with hypothesized Treg derivation have been reported.