Modulation of T Cell Function by Combination of Epitope Specific and Low Dose Anticytokine Therapy Controls Autoimmune Arthritis

Innate and adaptive immunity contribute to the pathogenesis of autoimmune arthritis by generating and maintaining inflammation, which leads to tissue damage. Current biological therapies target innate immunity, eminently by interfering with single pro-inflammatory cytokine pathways. This approach has shown excellent efficacy in a good proportion of patients with Rheumatoid Arthritis (RA), but is limited by cost and side effects. Adaptive immunity, particularly T cells with a regulatory function, plays a fundamental role in controlling inflammation in physiologic conditions. A growing body of evidence suggests that modulation of T cell function is impaired in autoimmunity. Restoration of such function could be of significant therapeutic value. We have recently demonstrated that epitope-specific therapy can restore modulation of T cell function in RA patients. Here, we tested the hypothesis that a combination of anti-cytokine and epitope-specific immunotherapy may facilitate the control of autoimmune inflammation by generating active T cell regulation. This novel combination of mucosal tolerization to a pathogenic T cell epitope and single low dose anti-TNFα was as therapeutically effective as full dose anti-TNFα treatment. Analysis of the underlying immunological mechanisms showed induction of T cell immune deviation

ATLAS Run 1 searches for direct pair production of third-generation squarks at the Large Hadron Collider

This paper reviews and extends searches for the direct pair production of the scalar supersymmetric partners of the top and bottom quarks in proton-proton collisions collected by the ATLAS collaboration during the LHC Run 1. Most of the analyses use 20 fb$^{-1}$ of collisions at a centre-of-mass energy of $\sqrt{s}$ = 8 TeV, although in some case an additional 4.7 fb$^{-1}$ of collision data at $\sqrt{s}$ = 7 TeV are used. New analyses are introduced to improve the sensitivity to specific regions of the model parameter space. Since no evidence of third-generation squarks is found, exclusion limits are derived by combining several analyses and are presented in both a simplified model framework, assuming simple decay chains, as well as within the context of more elaborate phenomenological supersymmetric models

Presence of the metabolic syndrome in testicular cancer patients and the relation with a SNP near IRS1

Background: Testicular cancer is the most common malignancy in men aged 20-40 years. The prognosis is relatively good; the current survival rate exceeds 90%. TC patients with disseminated disease receive chemotherapy. Due to the good survival a lot more cancer survivors are faced with the late effects of chemotherapy. A common late effect is the metabolic syndrome; in this study we want to investigate the prevalence, the relation with a single nucleotide polymorphism (SNP) near IRS1 (insulin receptor substrate 1) and body changes in relation to the metabolic syndrome. Methods and results: 246 patients were eligible for this study; follow-up data and IRS1 SNP data were collected from the genomic DNA of the patients. 1. For our first objective all 246 patients were analyzed and the metabolic syndrome was diagnosed in 30.9% of all patients with a median follow-up time of 6.3 years (range 1.4-20.2). 2. Our second primary objective was to test the relation between a SNP near IRS1 (rs2943650) and the presence of the metabolic syndrome during follow-up. All 246 patients were eligible for this part of the study. We found no significant relation between the presence of the metabolic syndrome and a SNP near IRS1 (rs2943650). The different genotypes were: TT (92/246), CT (117/246) and CC (37/246). 3. Our secondary objective was to detect changes in body composition and in relation with the development of the metabolic syndrome. For this analysis 141 patients were eligible because there were two follow-up visits available of these patients, so we could determine the presence of the metabolic syndrome at two moments. We studied three groups, the first group had no metabolic syndrome during follow-up, the second group developed it between the first and the second FU moment and the third group had the metabolic syndrome at both FU moments. Patients without the metabolic syndrome (N=95) had lower triglyceride and glucose levels in comparison with the groups who did develop metabolic syndrome during FU. Testosterone levels were also higher in the group without the metabolic syndrome. Another part of the study was to look at metabolic syndrome before chemotherapy. 104 patients were eligible for this part of the study because we presumed they did not have the metabolic syndrome before chemotherapy. We wanted to investigate predictors for the development of the metabolic syndrome by analyzing two groups who did not have metabolic syndrome at FU visit 1 but one group developed it at FU visit 2 and the other group did not. The data of the first follow-up visit were analyzed. We found that testosterone levels are significantly lower in the group that develops metabolic syndrome between FU 1 and 2. Insulin levels, BMI and waist circumference are increased in comparison with the group that does not develop the metabolic syndrome. Conclusion: Metabolic syndrome is a common late effect in patients with testicular cancer treated with chemotherapy. The prevalence is higher than in the general population and we investigated the relation between a known SNP near IRS1 and the presence of the metabolic syndrome, which we could not find. Body composition changes in patients developing the metabolic syndrome. Testosterone levels and insulin levels could be important predictors for the metabolic syndrome. Future research is needed to investigate the importance of these predictors and to investigate early treatment options.

Adoptive Transfer of Mandibular Lymphnode (MLN) T cells from Combination Therapy treated animals led to significant reduction of Adjuvant Arthritis (AA) in diseased animals, measured as Mean Arthritis Score as well as percentage of Disease Amelioration.

<div><p> <b>A.</b><i>Adoptive Transfer of T cells from Combination Therapy groups.</i> Adoptive Transfer Groups received 11×10⁶ Inguinal Lymphnode (ILN) cells, 13×10⁶ MLN cells, or 11×10⁶ spleen cells. Data represent Mean±SD. Disease induction and scoring was performed as described in the legend to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000087#pone-0000087-g001" target="_blank">Figure 1</a>.</p> <p> <b>B.</b><i>Percentage of Clinical Amelioration for each treatment group in AA rats</i>. The Area Under the Curve (AUC) of each individual treatment group was used to score the Clinical Amelioration (CA) of the distinct treatment groups. AUC was calculated using the curves originated by scoring the disease for the different treatment groups and plotted as percentage of CA with respect to the non-treated group. The non-treated group was considered as having an average percentage of disease = 100%. Formula is as follows: CA = 100 - %AUC.</p></div

Combination therapy as well as a full course of Etanercept treatment led to reduction of histological damage in the ankle joints.

<div><p>Joints were harvested on day 23 after the induction of arthritis.</p> <p>Formalin-fixed tissues were decalcified, and glass slides stained with H&E were prepared.</p> <p>Submitted tissue sections were examined by light microscopy and scored for severity of inflammation of the synovium, pannus formation, cartilage damage, bone marrow inflammation and periostal proliferation, with a maximum score of 4 per parameter.</p> <p>N = 3–4 per treatment group.</p> <p>H&E staining is shown of one rat per treatment, representative for the whole treatment group.</p> <p>A: Combination therapy; B: Etanercept 1×; C: Peptide Mt. 180-188 4× monotherapy; D: No treatment; E: Etanercept 3×.</p></div

The combination therapy of Etanercept and HSP60 180-188 led to an antigen specific increase of IL-10 and IL-4 production and up regulation of CTLA-4 expression in CD4+ T cells in draining Mandibular Lymphnodes (MLN).

<div><p>MLN were harvested on day 23 after the induction of arthritis.</p> <p>Cells were cultured for 72 hours with medium or antigen.</p> <p>Intracellular production of IL-4, IL-10, and expression of CTLA-4 were measured by FACS.</p> <p>Depicted are Mean Fluorescence Indexes (MFI) of MLN cells cultured with mycobacterial HSP60 peptide 180-188, of cells gated on CD4.</p> <p>Results are representative of one experiment.</p></div

Combination Therapy led to an increase in FOXP3 and IL-10 gene transcription in CD4+CD25+ cells, whereas it also led to and increase in IL-10 transcription in CD4+CD25− cells.

<div><p>TNFα transcription was abolished by the combination therapy as well as by a full course of Etanercept treatment.</p> <p>Results are expressed as the induction index (marker/housekeeping gene GAPDH) of HSP60 peptide 180-188 stimulation subtracted by media alone as measured by Real Time Quantitative PCR.</p> <p>IL-10 production Combination therapy vs. Etanercept treatment p = 0.002.</p> <p>N = 2–4 per treatment.</p></div

Combination therapy of Etanercept with mycobacterial heat shock protein 60 (HSP60) 180-188 led to significant reduction of Adjuvant Arthritis (AA).

<div><p>Arthritis was induced on day 0 with Complete Freund's Adjuvant (CFA).</p> <p>On day 9, rats were randomly divided into five treatment groups: three doses of Etanercept s.c. on day 9, 11, 13 (equivalent to a full course of Etanercept treatment); one dose of Etanercept on day 9; four doses of mycobacterial HSP60 peptide 180-188 on day 10, 13, 16, 19; combination treatment of one dose of Etanercept s.c. on day 9 followed by 180-188 nasally on day 10, 13, 16, 19; or no treatment (PBS).</p> <p>Arthritis scores were assessed every other day from day 8 onward.</p> <p>N = 15–18 rats per treatment group. Shown are mean arthritis scores.</p></div