R. De Palma

Anti-epidermal growth factor receptor monoclonal antibodies in cancer therapy

The epidermal growth factor receptor (EGFR) is a transmembrane tyrosine kinase receptor involved in the proliferation and survival of cancer cells. EGFR is the first molecular target against which monoclonal antibodies (mAb) have been developed for cancer therapy. Here we review the mechanisms underlying the effects of EGFR‐specific mAb in cancer therapy. The efficacy of EGFR‐specific mAb in cancer occurs thanks to inhibition of EGFR‐generated signalling; furthermore, the effects of antibodies on the immune system seem to play an important role in determining the overall anti‐tumour response. In this review, attention is focused on cetuximab and panitumumab, two mAb introduced recently into clinical practice for treatment of metastatic colorectal and head and neck cancer which target the external part of EGFR.

Th1/Th2 lymphocyte polarization in asthma

Asthma is a complex inflammatory disease of the lung characterized by variable airflow obstruction, bronchial hyperresponsiveness, and airway inflammation. Inflammation in asthma consists of airway infiltration by mast cells, lymphocytes, and eosinophils. There is accumulating evidence that CD4+ lymphocytes with a Th2‐cytokine pattern play a pivotal role in the pathogenesis of asthma. These cells orchestrate the recruitment and activation of the primary effector cells of the allergic response (mast cells and eosinophils), through the release of cytokines such as IL‐4, IL‐5, and IL‐13. Allergic inflammation is also implicated in airway epithelium changes, although the mechanisms by which inflammatory cells and, in particular, T cells interact with the epithelium are not completely clarified. This paper explores the role of T cells in the allergic inflammation of asthma.

Neopterin induces pro‐atherothrombotic phenotype in human coronary endothelial cells

Summary.  Background: Inflammation plays a pivotal role in atherothrombosis. Recent data indicate that serum levels of neopterin, a marker of inflammation and immune modulator secreted by monocytes/macrophages, are elevated in patients with acute coronary syndromes and seem to be a prognostic marker for major cardiovascular events. The aim of the present study was to determine whether neopterin might affect the thrombotic and atherosclerotic characteristics of human coronary artery endothelial cells (HCAECs). Methods and results: In HCAECs, neopterin induced TF‐mRNA transcription as demonstrated by real time polymerase chain reaction and expression of functionally active tissue factor (TF) as demonstrated by procoagulant activity assay, and of cellular adhesion molecules (CAMs) as demonstrated by FACS analysis, in a dose‐dependent fashion. These neopterin effects were prevented by lovastatin, a HMG‐CoA reductase inhibitor. Neopterin‐induced TF and CAMs expression was mediated by oxygen free radicals through the activation of the transcription factor, nuclear factor‐kappa B (NF‐κB), as demonstrated by electrophoretic mobility shift assay and by suppression of CAMs and TF expression by superoxide dismutase and by NF‐κB inhibitor, pyrrolidine‐dithio‐carbamate ammonium. Conclusions: These data indicate that neopterin exerts direct effects on HCAECs by promoting CAMs and TF expression and support the hypothesis that neopterin, besides representing a marker of inflammation, might be an effector molecule able to induce a pro‐atherothrombotic phenotype in cells of the coronary circulation.

Activation of protease‐activated receptor‐2 reduces airways inflammation in experimental allergic asthma

Background Proteinase‐activated receptors (PAR)‐2 are members of the family of G‐protein‐coupled receptors activated by proteases. These receptors are widely expressed in several tissues and in virtually all cells involved in rhinitis and asthma. In particular, proteinases activating PAR‐2 may affect airway functions and play a role in human diseases.

Human urotensin II induces tissue factor and cellular adhesion molecules expression in human coronary endothelial cells: an emerging role for urotensin II in cardiovascular disease

Summary.  Background: Human urotensin II is an 11‐aminoacid peptide with a controversial role in the human cardiovascular system. Indeed, urotensin effects on vascular reactivity and in heart failure are well documented, while its potential role in the pathophysiology of athero‐thrombosis is still unknown. This study investigates the effects of urotensin on tissue factor (TF) and VCAM‐1/ICAM‐1 expression in human coronary endothelial cells (HCAECs). Methods and results: Urotensin induced TF‐mRNA transcription as demonstrated by real time PCR and expression of TF that was functionally active as demonstrated by procoagulant activity assay. In addition, urotensin induced expression of VCAM‐1 and ICAM‐1 as demonstrated by FACS analysis. VCAM‐1 and ICAM‐1 were functionally active because they increased leukocyte adhesivity to HCAECs. Urotensin‐induced expression of TF and of VCAM‐1/ICAM‐1 was mediated through the Rho A‐activation of the transcription factor, NF‐κB, as demonstrated by EMSA. Indeed, lovastatin, an HMG‐CoA reductase inhibitor, by modulating the Rho activation, and NF‐κB inhibitors, suppressed the urotensin effects on TF and CAMs. Conclusions: Data of the present study, although in vitro, describe the close relationship existing between urotensin II and athero‐thrombosis, providing for the first time support for the view that this peptide might have not only vasoactive functions but it might be an effector molecule able to induce a pro‐atherothrombotic phenotype in cells of the coronary circulation. Although future studies are required to clarify whether these mechanisms are also important in the clinical setting, this report supports an emerging new role for urotensin II in the pathogenesis and progression of cardiovascular disease.

Peripheral T lymphocytes from patients with early systemic sclerosis co‐cultured with autologous fibroblasts undergo an oligoclonal expansion similar to that occurring in the skin

In recent years several reports have suggested that T cells may have a role in systemic sclerosis (SSc). The aim of our study was to investigate the dynamics of T cell repertoire in early SSc disease analysing a target organ, the skin, and the peripheral blood. To date, indeed, it is not clear if T cell expansions found in SSc reflect a general activation or result from specific antigen stimulation in the target organs. This is an important point to assess in order to characterize the role of T cells in the development of SSc. To address these questions we studied T cell repertoire by CDR3 length analysis in skin biopsies and peripheral blood obtained from patients affected by SSc and we found that a skewed T cell repertoire was present only in the biopsies. In order to characterize more effectively the meaning of these data, we performed co‐cultures using fibroblasts and peripheral blood mononuclear cells (PBMCs) obtained from SSc patients. These experiments showed that same T cell expansions were detectable in the skin of SSc patients and in the cultures of PBMCs and autologous fibroblasts of the patients but not in their peripheral blood. Taken together, these data suggest that fibroblasts trigger specific T cell expansions in the early phase of SSc.

Updating clinical recommendations for breast, colorectal and lung cancer treatments: an opportunity to improve methodology and clinical relevance

BACKGROUND clinical guidelines can improve quality of care summarising available knowledge and proposing recommendations for health care decisions. Being up to date is one of their quality requisites. Little experience is available on when and how guidelines should be updated. We report on the update process of evidence-based clinical recommendations on anticancer drugs. METHODS three multidisciplinary panels, supported by methodology experts, updated the recommendations. The methodologists were in charge of the qualitative and quantitative synthesis of the evidence. The panels were responsible for the final decision about risk/benefit profile of the drugs and strength of the recommendations. The GRADE (Grading of Recommendations Assessment, Development and Evaluation) approach was used. RESULTS six recommendations out of 15 were completely updated in 8 months time. In four cases, the strength of the recommendation changed; in two of them, we moved from a weak to a strong positive one. Despite the increased certainty about the positive risk/benefit profile, this was translated in a change in the strength of the recommendation only in one case out of three. Three recommendations were refined making them more clinically specific. CONCLUSIONS accumulation of evidence is an opportunity for guideline panels to refine methodological rigour, clinical relevance and to foster consensus on recommendations. This requires time and resource investments.

Use of altered peptide ligands to modulate immune responses as a possible immunotherapy for allergies

Allergies are dramatically increasing in prevalence, and the management of these diseases is a heavy burden on the health‐care systems of developed countries. In recent years, many efforts have been made to improve the therapy of allergies and to develop new approaches for immunotherapy. Here we briefly review the use of peptides to modulate T‐cell responses to allergens. We focus mainly on the possibility of using altered peptide ligands (APLs), i.e., peptides tailored on immunodominant T epitopes and bearing a single amino‐acid substitution, as a tool to modulate immune responses to allergens. These peptides may be recognized by the specific T cells triggered by the agonist peptides, but they are unable to elicit T‐cell responses; thus, they could be ideal candidates to modulate immune responses to allergens. The availability of these peptides could allow new approaches for immunotherapies.

FRI0522 Peripheral Blood Mononuclear Cells Co-Cultured with Autologous Skin Fibroblasts Up-Regulate IL-17A and Play Anti-Fibrotic Effects in Systemic Sclerosis

Background IL-17A has been recently implicated in the pathogenesis of systemic sclerosis (SSc). Objectives Therefore, we explored its expression and effects in peripheral blood mononuclear cells (PBMCs) co-cultured with autologous skin fibroblasts. Methods PBMCs and autologous skin fibroblasts from 5 patients with early (disease duration <3 years) diffuse (dc) SSc were co-cultured in presence of IL-2 20U/ml in a 1:10 ratio. Separate cultures of PBMCs with IL-2 20U/ml and unstimulated fibroblasts were used as controls. The expression of IL17A, IL1B, IL4, and TGFB1 mRNA was analyzed in co-cultured and control PBMCs after 10 days by real-time PCR. The expression of IL17RA, CCL2, CCL3, CXCL1, COL1A1, COL3A1, CTGF, TGFBR2, and SMAD3 mRNA was analyzed in co-cultured and control fibroblasts. Chemokine production was further investigated at the protein level in culture surnatants by multiplex suspension fluorescence-based immunoassay. Results Real-time PCR analysis showed an increased expression of IL17A in co-cultured PBMCs by 11.5 fold (p<0.01). No significant difference was found in the expression of IL1B, IL4, and TGFB1 between co-cultured and control PBMCs. Consistently with IL17A up-regulation in co-cultured PBMCs, mRNA levels of IL17RA were increased by 4.3 fold in co-cultured fibroblasts (p<0.05). In order to clarify whether this up-regulation ensued in the formation of a functional receptor, we analyzed the expression of CCL2, CCL3, and CXCL1, which are IL-17A target genes, in co-cultured fibroblasts. Indeed, CCL2, CCL3, and CXCL1 were up-regulated by 11.9 fold, 773.26 fold, and 29 fold respectively (p<0.05). This induction was confirmed at the protein level in surnatants from co-cultures (CCL-2 25014 pg/ml; CCL-3 2227 pg/ml; CXCL-1 605.8 pg/ml) compared to control PBMCs (CCL-2 2821 pg/ml; CCL-3 199.1 pg/ml; CXCL-1 2.4 pg/ml) and control fibroblasts (CCL-2 2894 pg/ml; CCL-3 21.56 pg/ml; CXCL-1 3.1 pg/ml) (p<0.05). Lastly, we investigated the effects of co-cultured PBMCs on the expression of pro-fibrotic genes in fibroblasts. We found that COL1A1, COL3A1, and CTGF mRNAs were down-regulated by 0.33 fold, 0.24 fold, and 0.31 fold respectively (p<0.05). In addition, we also found a down-regulation by 0.78 fold of TGFBR2 and by 0.79 of SMAD3, suggesting that co-cultured PBMCs may interfere with the TGF-beta pathway hyperactivity in SSc fibroblasts. Conclusions Here we show for the first time that PBMCs from early dcSSc co-cultured with autologous skin fibroblasts over-express IL17A and exert anti-fibrotic effects in vitro. The simultaneous up-regulation of IL17RA and IL-17A target genes in the co-cultured fibroblasts suggests that IL-17A pathway is active in early dcSSc fibroblasts. These findings are paralleled by the down-regulation of TGF-beta signalling components. We previously showed that in co-cultures performed with PBMCs and autologous skin fibroblasts from early dcSSc patients T cells are expanded and kill the autologous fibroblasts. Taken together, these novel data support the hypothesis that immune system may be primarily aimed to control fibroblast activation in the early phases of the disease, potentially opening new therapeutic approaches in SSc. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.5910