Value through Innovation22 December 2014

Scientific Publications

Boehringer Ingelheim offers an overview of scientific publications on the following pages. This overview represents all publications of the last three years (YTD) where employees of Boehringer Ingelheim worldwide were involved.

30 publications regarding Immunology
  • Author:
    El-Far M; Pellerin C; Pilote L; Fortin J-F; Lessard IA D; Peretz Y; Wardrop E; Salois P; Bethell RC; Cordingley MG; Kukolj G
    Title:
    CD160 isoforms and regulation of CD4 and CD8 T-cell responses
    Source:
    J Transl Med 12 (1) art. no. 217 (2014)
    Abstract:
    Background: Coexpression of CD160 and PD-1 on HIV-specific CD8+ T-cells defines a highly exhausted T-cell subset. CD160 binds to Herpes Virus Entry Mediator (HVEM) and blocking this interaction with HVEM antibodies reverses T-cell exhaustion. As HVEM binds both inhibitory and activatory receptors, our aim in the current study was to assess the impact of CD160-specific antibodies on the enhancement of T-cell activation.Methods: Expression of the two CD160 isoforms; glycosylphosphatidylinositol-anchored (CD160-GPI) and the transmembrane isoforms (CD160-TM) was assessed in CD4 and CD8 primary T-cells by quantitative RT-PCR and Flow-cytometry. Binding of these isoforms to HVEM ligand and the differential capacities of CD160 and HVEM specific antibodies to inhibit this binding were further evaluated using a Time-Resolved Fluorescence assay (TRF). The impact of both CD160 and HVEM specific antibodies on enhancing T-cell functionality upon antigenic stimulation was performed in comparative ex vivo studies using primary cells from HIV-infected subjects stimulated with HIV antigens in the presence or absence of blocking antibodies to the key inhibitory receptor PD-1.Results: We first show that both CD160 isoforms, CD160-GPI and CD160-TM, were expressed in human primary CD4+ and CD8+ T-cells. The two isoforms were also recognized by the HVEM ligand, although this binding was less pronounced with the CD160-TM isoform. Mechanistic studies revealed that although HVEM specific antibodies blocked its binding to CD160-GPI, surprisingly, these antibodies enhanced HVEM binding to CD160-TM, suggesting that potential antibody-mediated HVEM multimerization and/or induced conformational changes may be required for optimal CD160-TM binding. Triggering of CD160-GPI over-expressed on Jurkat cells with either bead-bound HVEM-Fc or anti-CD160 monoclonal antibodies enhanced cell activation, consistent with a positive co-stimulatory role for CD160-GPI. However, CD160-TM did not respond to this stimulation, likely due to the lack of optimal HVEM binding. Finally, ex vivo assays using PBMCs from HIV viremic subjects showed that the use of CD160-GPI-specific antibodies combined with blockade of PD-1 synergistically enhanced the proliferation of HIV-1 specific CD8+ T-cells upon antigenic stimulation.Conclusions: Antibodies targeting CD160-GPI complement the blockade of PD-1 to enhance HIV-specific T-cell responses and warrant further investigation in the development of novel immunotherapeutic approaches.
  • Author:
    .krnjug I; Rueckert C; Libanova R; Lienenklaus S; Weiss S; Guzmán CA
    Title:
    The mucosal adjuvant cyclic di-AMP exerts immune stimulatory effects on dendritic cells and macrophages
    Source:
    PLoS ONE 9 (4) art.no.e95728 (2014)
    Abstract:
    The cyclic di-nucleotide bis-(3.,5.)-cyclic dimeric adenosine monophosphate (c-di-AMP) is a candidate mucosal adjuvant with proven efficacy in preclinical models. It was shown to promote specific humoral and cellular immune responses following mucosal administration. To date, there is only fragmentary knowledge on the cellular and molecular mode of action of c-di-AMP. Here, we report on the identification of dendritic cells and macrophages as target cells of c-di-AMP. We show that c-di-AMP induces the cell surface up-regulation of T cell co-stimulatory molecules as well as the production of interferon-.. Those responses were characterized by in vitro experiments with murine and human immune cells and in vivo studies in mice. Analyses of dendritic cell subsets revealed conventional dendritic cells as principal responders to stimulation by c-di-AMP. We discuss the impact of the reported antigen presenting cell activation on the previously observed adjuvant effects of c-di-AMP in mouse immunization studies. © 2014 .krnjug et al.
  • Author:
    Profita M; Albano GD; Riccobono L; Di Sano C; Montalbano AM; Gagliardo R; Anzalone G; Bonanno A; Pieper MP; Gjomarkaj M
    Title:
    Increased levels of Th17 cells are associated with non-neuronal acetylcholine in COPD patients
    Source:
    Immunobiology 219 (5), 392-401 (2014)
    Abstract:
    T-lymphocytes, including Th17-cells and T-cells expressing acetylcholine (ACh), are key components of systemic inflammation in chronic obstructive pulmonary disease (COPD). We investigated whether ACh promotes Th17 cells in COPD.ACh, IL-17A, IL-22, ROR.t, FOXP3 expression and AChIL-17A, AChIL-22, AChROR.t coexpression was evaluated in peripheral blood mononuclear cells (PBMC) from COPD patients (n=16), healthy smokers (HS) (n=12) and healthy control subjects (HC) (n=13) (cultured for 48h with PMA) by flow cytometry. Furthermore, we studied the effect of Tiotropium (Spiriva®) (100nM) and Olodaterol (1nM) alone or in combination, and of hemicholinium-3 (50.M) on AChIL-17A, AChIL-22, AChROR.t, and FOXP3 expression in CD3+PBT-cells of PBMC from COPD patients (n=6) cultured for 48h with PMA.CD3+PBT-cells expressing ACh, IL-17A, IL-22 and ROR.t together with CD3+PBT-cells co-expressing AChIL-17A, AChIL-22 and AChROR.t were significantly increased in COPD patients compared to HC and HS subjects with higher levels in HS than in HC without a significant difference. CD3+FOXP3+PBT-cells were increased in HS than in HC and COPD. Tiotropium and Olodaterol reduced the percentage of CD3+PBT-cells co-expressing AChIL-17A, AChIL-22, and AChROR.t while increased the CD3+FOXP3+PBT-cells in PBMC from COPD patients, cultured in vitro for 48h, with an additive effect when used in combination. Hemicholnium-3 reduced the percentage of ACh+IL-17A+, ACh+IL-22+, and ACh+ROR.t+ while it did not affect FOXP3+ expression in CD3+PBT-cells from cultured PBMC from COPD patients.We concluded that ACh might promote the increased levels of Th17-cells in systemic inflammation of COPD. Long-acting .2-agonists and anticholinergic drugs might contribute to control this event. © 2014 Elsevier GmbH.
  • Author:
    Krug N; Gupta A; Badorrek P; Koenen R; Mueller M; Pivovarova A; Hilbert J; Wetzel K; Hohlfeld JM; Wood C
    Title:
    Efficacy of the oral chemoattractant receptor homologous molecule on T H2 cells antagonist BI 671800 in patients with seasonal allergic rhinitis
    Source:
    J Allergy Clin Immunol 133 (2), 414-419 e8 (2014)
    Abstract:
    Background The inflammatory response in patients with seasonal allergic rhinitis (SAR) is partly mediated by the prostaglandin D2 receptor chemoattractant receptor homologous molecule on TH2 cells (CRTH2). Objective We sought to investigate the efficacy and safety of the oral CRTH2 antagonist BI 671800 (50, 200, and 400 mg twice daily), fluticasone propionate nasal spray (200 .g once daily), or oral montelukast (10 mg once daily) administered for 2 weeks in patients with SAR. Methods In this randomized, double-blind, placebo-controlled, partial-crossover study, participants aged 18 to 65 years with a positive skin prick test to Dactylis glomerata pollen were exposed to out-of-season allergen in the environmental challenge chamber for 6 hours. The primary efficacy variable was the total nasal symptom score assessed as the area under the curve (AUC)0-6h. Results In total, 146 patients (63.7% male; mean age, 36.1 years) were randomized. The adjusted mean total nasal symptom score AUC0-6h was significantly reduced versus placebo with 200 mg of BI 671800 (absolute difference, -0.85; percentage difference, -17%; P =.0026), montelukast (absolute difference, -0.74; percentage difference, -15%; P =.0115), and fluticasone propionate (absolute difference, -1.64; percentage difference, -33%; P <.0001). Compared with placebo, BI 671800 significantly reduced nasal eosinophil values (P <.05 for all doses), significantly inhibited nasal inflammatory cytokine levels (IL-4 and eotaxin, P <.05; 200 mg twice daily), and induced a dose-related reduction in ex vivo prostaglandin D2-mediated eosinophil shape change. Conclusion Two hundred milligrams of BI 671800 twice daily demonstrated efficacy in treating SAR symptoms induced by environmental challenge chamber allergen exposure and had a favorable safety profile. © 2013 American Academy of Allergy, Asthma & Immunology
  • Author:
    Profita M; Albano GD; Riccobono L; Di Sano C; Montalbano AM; Gagliardo R; Anzalone G; Bonanno A; Pieper MP; Gjomarkaj M
    Title:
    Increased levels of Th17 cells are associated with non-neuronal acetylcholine in COPD patients
    Source:
    Immunobiology Article in press (2014)
    Abstract:
    T-lymphocytes, including Th17-cells and T-cells expressing acetylcholine (ACh), are key components of systemic inflammation in chronic obstructive pulmonary disease (COPD). We investigated whether ACh promotes Th17 cells in COPD. ACh, IL-17A, IL-22, ROR.t, FOXP3 expression and AChIL-17A, AChIL-22, AChROR.t coexpression was evaluated in peripheral blood mononuclear cells (PBMC) from COPD patients (n = 16), healthy smokers (HS) (n = 12) and healthy control subjects (HC) (n = 13) (cultured for 48 h with PMA) by flow cytometry. Furthermore, we studied the effect of Tiotropium (Spiriva®) (100 nM) and Olodaterol (1 nM) alone or in combination, and of hemicholinium-3 (50 .M) on AChIL-17A, AChIL-22, AChROR.t, and FOXP3 expression in CD3+PBT-cells of PBMC from COPD patients (n = 6) cultured for 48 h with PMA. CD3+PBT-cells expressing ACh, IL-17A, IL-22 and ROR.t together with CD3+PBT-cells co-expressing AChIL-17A, AChIL-22 and AChROR.t were significantly increased in COPD patients compared to HC and HS subjects with higher levels in HS than in HC without a significant difference. CD3+FOXP3+PBT-cells were increased in HS than in HC and COPD. Tiotropium and Olodaterol reduced the percentage of CD3+PBT-cells co-expressing AChIL-17A, AChIL-22, and AChROR.t while increased the CD3+FOXP3+PBT-cells in PBMC from COPD patients, cultured in vitro for 48 h, with an additive effect when used in combination. Hemicholnium-3 reduced the percentage of ACh+IL-17A+, ACh+IL-22+, and ACh+ROR.t+ while it did not affect FOXP3+ expression in CD3+PBT-cells from cultured PBMC from COPD patients. We concluded that ACh might promote the increased levels of Th17-cells in systemic inflammation of COPD. Long-acting .2-agonists and anticholinergic drugs might contribute to control this event. © 2014.
  • Author:
    Teixeira LH; Tararam CA; Lasaro MO; Camacho AGA; Ersching J; Leal MT; Herrera S; Bruna-Romero O; Soares IS; Nussenzweig RS; Ertl HCJ; Nussenzweig V; Rodrigues MM
    Title:
    Immunogenicity of a prime-boost vaccine containing the circumsporozoite proteins of plasmodium vivax in rodents
    Source:
    Infect Immun 82 (2), 793-807 (2014)
    Abstract:
    Plasmodium vivax is the most widespread and the second most prevalent malaria-causing species in the world. Current measures used to control the transmission of this disease would benefit from the development of an efficacious vaccine.In the case of the deadly parasite P. falciparum, the recombinant RTS,S vaccine containing the circumsporozoite antigen (CSP) consistently protects 30 to 50% of human volunteers against infection and is undergoing phase III clinical trials in Africa with similar efficacy. These findings encouraged us to develop a P. vivax vaccine containing the three circulating allelic forms of P. vivax CSP. Toward this goal, we generated three recombinant bacterial proteins representing the CSP alleles, as well as a hybrid polypeptide called PvCSP-All-CSP-epitopes. This hybrid contains the conserved N and C termini of P. vivax CSP and the three variant repeat domains in tandem. We also generated simian and human recombinant replication-defective adenovirus vectors expressing PvCSP-All-CSP-epitopes. Mice immunized with the mixture of recombinantproteins in a formulation containing the adjuvant poly(I.C) developed high and long-lasting serum IgG titers comparable to those elicited by proteins emulsified in complete Freund's adjuvant. Antibody titers were similar in mice immunized with homologous (protein-protein) and heterologous (adenovirus-protein) vaccine regimens. The antibodies recognized the three allelic forms of CSP, reacted to the repeated and nonrepeated regions of CSP, and recognized sporozoites expressing the alleles VK210 and VK247. The vaccine formulations described in this work should be useful for the further development of an anti-P. vivax vaccine. © 2014, American Society for Microbiology.
  • Author:
    Krug N; Gupta A; Badorrek P; Koenen R; Mueller M; Pivovarova A; Hilbert J; Wetzel K; Hohlfeld JM; Wood C
    Title:
    Efficacy of the oral chemoattractant receptor homologous molecule on TH2 cells antagonist BI 671800 in patients with seasonal allergic rhinitis
    Source:
    J Allergy Clin Immunol Article in press (2013)
    Abstract:
    Background: The inflammatory response in patients with seasonal allergic rhinitis (SAR) is partly mediated by the prostaglandin D2 receptor chemoattractant receptor homologous molecule on TH2 cells (CRTH2). Objective: We sought to investigate the efficacy and safety of the oral CRTH2 antagonist BI 671800 (50, 200, and 400 mg twice daily), fluticasone propionate nasal spray (200 .g once daily), or oral montelukast (10 mg once daily) administered for 2 weeks in patients with SAR. Methods: In this randomized, double-blind, placebo-controlled, partial-crossover study, participants aged 18 to 65 years with a positive skin prick test to Dactylis glomerata pollen were exposed to out-of-season allergen in the environmental challenge chamber for 6 hours. The primary efficacy variable was the total nasal symptom score assessed as the area under the curve (AUC)0-6h. Results: In total, 146 patients (63.7% male; mean age, 36.1 years) were randomized. The adjusted mean total nasal symptom score AUC0-6h was significantly reduced versus placebo with 200 mg of BI 671800 (absolute difference, -0.85; percentage difference, -17%; P = .0026), montelukast (absolute difference, -0.74; percentage difference, -15%; P = .0115), and fluticasone propionate (absolute difference, -1.64; percentage difference, -33%; P < .0001). Compared with placebo, BI 671800 significantly reduced nasal eosinophil values (P < .05 for all doses), significantly inhibited nasal inflammatory cytokine levels (IL-4 and eotaxin, P < .05; 200 mg twice daily), and induced a dose-related reduction in ex vivo prostaglandin D2-mediated eosinophil shape change. Conclusion: Two hundred milligrams of BI 671800 twice daily demonstrated efficacy in treating SAR symptoms induced by environmental challenge chamber allergen exposure and had a favorable safety profile. © 2013 American Academy of Allergy, Asthma & Immunology
  • Author:
    Noyes A; Boesch A; Godavarti R; Titchener-Hooker N; Coffman J; Mukhopadhyay T
    Title:
    High throughput quantification of capsular polysaccharides for multivalent vaccines using precipitation with a cationic surfactant
    Source:
    Vaccine 31 (48), 5659-5665 (2013)
    Abstract:
    The increasing requirement for multivalent vaccines containing diverse capsular polysaccharides has created an unmet need for a fast and straightforward assay for polysaccharide titer. We describe a novel and robust assay for the quantitation of anionic capsular polysaccharides. The binding of hexadecyltrimethyammonium bromide (Hb) to anionic capsular polysaccharides results in a precipitation reaction wherein the suspension turbidity is proportional to polysaccharide titer. The turbidity can be quickly measured as absorbance across a range of wavelengths that resolve scattering light. Carbohydrates comprised of repeating units of one to seven monosaccharides with phosphodiester groups, uronic acids, and sialic acids all reacted strongly and there does not appear to be specificity with respect to the particular anionic moiety. The assay is compatible with an array of common buffers across a pH range of 3.0-8.75 and with NaCl concentration exceeding 400. mM. Interference from DNA can be eliminated with a short incubation step with DNase. With these treatments, the assay has been employed in samples as complex as fermentation broth. A two-log dynamic range has been established with a mean relative standard deviation less than 10% across this range although inferior performance has been observed in fermentation broth.The precipitation assay enables the rapid quantitation of anionic polysaccharides. The resulting procedure can robustly measure the titer of myriad anionic capsular polysaccharides (CPS) in 96 samples in less than 30. min using low toxicity reagents and routine laboratory equipment. This development will greatly reduce the effort required to measure polysaccharide titer and yield during process development of polysaccharide vaccines. © 2013 The Authors.
  • Author:
    Bethell R; Scherer J; Witvrouw M; Paquet A; Coakley E; Hall D
    Title:
    Short communication: Phenotypic protease inhibitor resistance and cross-resistance in the clinic from 2006 to 2008 and mutational prevalences in HIV from patients with discordant tipranavir and darunavir susceptibility phenotypes.
    Source:
    AIDS Res Hum Retroviruses 28 (9), 1019-1024 (2012)
    Abstract:
    To test tipranavir (TPV) or darunavir (DRV) as treatment options for patients with phenotypic resistance to protease inhibitors (Pis), including lopinavir, saquinavir, atazanavir, and fosamprenavir, the PhenoSense GT database was analyzed for susceptibility to DRV or TPV among Pl-resistant isolates. The Monogram Bioseiences HIV database (South San Francisco, CA) containing 7775 clinical isolates (2006-2008) not susceptible to at least one first-generation PI was analyzed. Phenotypic responses [resistant (R), partially susceptible (PS), or susceptible (S)] were defined by upper and lower clinical cut-offs to each PI. Genetypes were screened for amino acid substitutions associated with TPV-R/DRV-S and TPV-S/DRV-R phenotypes. ln all, 4.9% (378) of isolates were resistant to all six Pis and 31.0% (2407) were resistant to none. Among isolates resistant to all four first-generation Pis, DRV resistance increased from 21.2% to 41.9% from 2006 to 2008, respectively, and resistance to TPV remained steady (53.9 to 57.3%, respectively). Higher prevalence Substitutions in DRV-S/TPV-R isolates versus DRV-R/TPV-S isolates, respectively, were 82L/T (44.4% vs. 0%) and 830 (5.8% vs. 0%). Higher prevalence Substitutions in DRV-R/TPV-S virus were 50V (0.0% vs. 28.9%), 54L (1.0% vs. 36.1%), and 76V (0.4% vs. 15.5%). Mutations to help predict discordant susceptibility to DRV and TPV in isolates with reduced susceptibility to other Pis were identified. DRV resistance mutations associated with improved virologic response to TPV were more prevalent in DRV-R/TPV-S isolates. TPV resistance mutations were more prevalent in TPV-R and DRV-S isolates. These results confirm the impact of genotype on phenotype, illustrating how HIV genotype and phenotype data assist regimen optimization.
  • Author:
    Engel Michael; Schmidt Hendrick; Moroni-Zentgraf Petra
    Title:
    Efficacy of tiotropium in patients with asthma in relation to allergic status.
    Source:
    J Allergy Clin Immunol 131 (2) (1), AB1 (2013)
    Abstract:
    RATIONALE: The response of patients with asthma to some medications may be influenced by their allergic status. Tiotropium, a long-acting anticholinergic, has demonstrated efficacy and safety in patients with asthma (Kerstjens et al. NEJM 2012), but the relationship between response and patient allergic status has not been published in detail. METHODS: Analysis of prespecified subgroups was performed using data from two replicate, 48-week, placebo-controlled trials (N5912) conducted in patients with asthma on at least ICS+LABA randomized to adding tiotropium 5.mu.g Respimat or placebo once daily. The subgroup of patients with potentially allergic asthma was identified using three criteria: total serum IgE, blood eosinophils, or clinician judgment (CJ). Allergic status was positive if serum IgE was >430 .mu.g/L, blood eosinophils were >0.6.times.10 /L, or CJ was .mchlt.yes..mchgt. RESULTS: Peak FEV1 improved with tiotropium irrespective of allergic status in Trial 1 (IgE [subgroup3treatment interaction, P=0.86] and eosinophils [P=0.46]) and in Trial 2 (IgE [P=0.98]; eosinophils P=0.18]; and CJ [P=0.29]). Predose (trough) FEV1improved with tiotropium compared with placebo, irrespective of allergic status, across all three criteria (Trial 1 [IgE, P=0.85; eosinophils, P=0.83; and CJ, P=0.15]; Trial 2 [IgE, P=0.58; eosinophils, P=0.38; and CJ, P=0.85]). Pooled data analyses revealed that time to first severe asthma exacerbation and time to first asthma worsening were both increased with tiotropium compared with placebo, regardless of allergic status, based on the three criteria. Overall, adverse events were balanced. CONCLUSIONS: Tiotropium improved lung function and asthma control in patients with poorly controlled asthma despite treatment with ICS+LABA, irrespective of their allergic status.
  • Author:
    Quester I; Peters M; Klein S; Deschl U; Wohlsein P
    Title:
    Case report: Infections with Toxoplasma gondii and Frenkelia sp. in chinchillas (Chinchilla lanigera) [Fallbericht: Infektionen mit Toxoplasma gondii und Frenkelia sp. bei Chinchillas (Chinchilla lanigera)]
    Source:
    Prakt Tierarzt 93 (6), 494-502 (2012)
    Abstract:
    A four to five years old male chinchilla died after a few days of reduced food intake. A one year old female chinchilla was also submitted for necropsy. It originated from a different breeding herd, where twelve dams and four offspring had died within three weeks with apathy and head tilt. Histologically, both animals suffered from a protozoal necrotising encephalitis with intra-and perilesional protozoal microorganisms that were found free or aggregated in cysts of 40-80 .m. In addition, in the brain of the dam numerous protozoal cysts up to 250 .m in diameter without inflammatory response were detected suggestive of Frenkelia microti cysts. In several internal organs of the male including heart, lung, gastro-intestinal tract, liver, pancreas, musculature, testis, adrenal and thyroid glands inflammatory changes were present. Placenta and liver of the dam also showed inflammation. Immunohistology revealed Toxoplasma gondii in both animals. In chinchillas with central nervous signs or sudden death toxoplasmosis should be considered as differential diagnosis. © Schlütersche Verlagsgesellschaft mbH & Co. KG.
  • Author:
    Miriyala S; Panchatcharam M; Ramanujam M; Puvanakrishnan R
    Title:
    A novel tetrapeptide derivative exhibits in vitro inhibition of neutrophil-derived reactive oxygen species and lysosomal enzymes release
    Source:
    Oxidative Med Cell Longevity art no 853210 (2013)
    Abstract:
    Neutrophil infiltration plays a major role in the pathogenesis of myocardial injury. Oxidative injury is suggested to be a central mechanism of the cellular damage after acute myocardial infarction. This study is pertained to the prognostic role of a tetrapeptide derivative PEP1261 (BOC-Lys(BOC)-Arg- Asp-Ser(tBu)-OtBU), a peptide sequence (39-42) of lactoferrin, studied in the modulation of neutrophil functions in vitro by measuring the reactive oxygen species (ROS) generation, lysosomal enzymes release, and enhanced expression of C proteins. The groundwork experimentation was concerned with the isolation of neutrophils from the normal and acute myocardial infarct rats to find out the efficacy of PEP1261 in the presence of a powerful neutrophil stimulant, phorbol 12-myristate 13 acetate (PMA). Stimulation of neutrophils with PMA resulted in an oxidative burst of superoxide anion and enhanced release of lysosomal enzymes and expression of complement proteins. The present study further demonstrated that the free radicals increase the complement factors in the neutrophils confirming the role of ROS. PEP1261 treatment significantly reduced the levels of superoxide anion and inhibited the release of lysosomal enzymes in the stimulated control and infarct rat neutrophils. This study demonstrated that PEP1261 significantly inhibited the effect on the ROS generation as well as the mRNA synthesis and expression of the complement factors in neutrophils isolated from infarct heart. © 2013 Sumitra Miriyala et al.
  • Author:
    Miriyala S; Panchatcharam M; Landar A; Ramanujam M; Chatterjee S; Muthuswamy A
    Title:
    The janus of oxidative stress signaling in different pathophysiological conditions
    Source:
    Oxidative Med Cell Longevity art no 708796 (2013)
    Abstract:
    no abstract available
  • Author:
    Wu H S; Lunter A-K; Spillner E; Schmitz N; Heider K-H; Zeis M
    Title:
    A Novel Anti-CD37 antibody shows superior antibody dependent cellualar cytotoxic activity (ADCC) compared to Rituximab against patient-derived Lymphoma cells
    Source:
    DGHO Jahrestagung, Wien 18-22.10. 2013
  • Author:
    Sathish JG; Sethu S; Bielsky M-C; De Haan L; French NS; Govindappa K; Green J; Griffiths CEM; Holgate S; Jones D; Kimber I; Moggs J; Naisbitt DJ; Pirmohamed M; Reichmann G; Sims J; Subramanyam M; Todd MD; Van Der Laan JW; Weaver RJ; Park BK
    Title:
    Challenges and approaches for the development of safer immunomodulatory biologics
    Source:
    Nat Rev Drug Discov 12 (4), 306-324 (2013)
    Abstract:
    Immunomodulatory biologics, which render their therapeutic effects by modulating or harnessing immune responses, have proven their therapeutic utility in several complex conditions including cancer and autoimmune diseases. However, unwanted adverse reactions-including serious infections, malignancy, cytokine release syndrome, anaphylaxis and hypersensitivity as well as immunogenicity-pose a challenge to the development of new (and safer) immunomodulatory biologics. In this article, we assess the safety issues associated with immunomodulatory biologics and discuss the current approaches for predicting and mitigating adverse reactions associated with their use. We also outline how these approaches can inform the development of safer immunomodulatory biologics.
  • Author:
    Freebern W J; Bigwarfe T J; Price K D; Haggerty H G
    Title:
    Methods: Implementation of in vitro and ex vivo phagocytosis and respiratory burst function assessments in safety testing
    Source:
    J Immunotoxicol 10 (1), 106-117 (2013)
    Abstract:
    Abstract Functional innate immune assessments, including phagocytosis and respiratory burst, are at the forefront of immunotoxicology evaluation in pre-clinical animal species. Although in the clinic and in academic science, phagocytosis, and respiratory burst assessments have been reported for over two decades, the implementation of phagocytosis and respiratory burst analyses in toxicology safety programs is just recently gaining publicity. Discussed herein are general methods, both microtiter plate-based and flow cytometric-based, for assessing phagocytosis and respiratory burst in pre-clinical species including mouse, rat, dog, and monkey. This methods-centric discussion includes a review of technologies and descriptions of method applications, with examples of results from analyses testing reported inhibitors (rottlerin, wortmannin, and SB203580) of phagocytosis and respiratory burst. Justification of implementation, strategic experimental design planning, and feasibility aspects of evaluating test article effects on phagocytosis and respiratory burst function are described within the context of a case study. The case study involves investigation of the effects of a small molecule p38 kinase inhibitor, BMS-582949, on phagocytosis and respiratory burst functions in rat and monkey neutrophils and monocytes in vitro, as well as ex vivo in these innate immune cells from monkeys administered BMS-582949 during a 1-week repeat dose investigative study. The results of the in vitro and ex vivo assessments demonstrated that BMS-582949 inhibited phagocytosis and respiratory burst. These findings correlated with incidences of opportunistic infections observed in rat and monkey toxicity studies. © 2013 Informa Healthcare USA, Inc.
  • Author:
    Tilp, C.; Kapur, V.; Loging, W.; Erb, K.J.
    Title:
    Prerequisites for the pharmaceutical industry to develop and commercialise helminths and helminth-derived product therapy
    Source:
    Int. J. Parasitol 43 (3-4), 319-325 (2013)
    Abstract:
    During the past 10. years, immunologists, epidemiologists and parasitologists have made many new exciting discoveries in the field of helminth-mediated immune regulation. In addition, many animal experiments have shown that certain helminths or products derived from helminths can protect mice from developing allergic or autoimmune disease. Some clinical trials utilising Trichuris suis or Necator americanus for the treatment of allergic disorders and inflammatory bowel disease have been conducted. The outcomes of these trials suggest that they may be used to treat these disorders. However, to date no helminth therapy is routinely being applied to patients and no helminth-derived product therapy has been developed. In order to bring new drugs to the market and shoulder the enormous costs involved in developing such therapies, pharmaceutical companies need to be involved. However, currently the resources from the pharmaceutical industry devoted to this concept are relatively small and there are good reasons why the industry may have been reluctant to invest in developing these types of therapies. In this review article, the hurdles that must be overcome before the pharmaceutical industry might invest in these novel therapies are outlined. © 2013 Australian Society for Parasitology Inc.
  • Author:
    Ernsting A; Gellert P; Schneider M; Lippke S
    Title:
    A mediator model to predict: workplace influenza vaccination behaviour - an application of the health action process approach.
    Source:
    Psychol Health Article in Press (2012)
  • Author:
    Moss N; Xiong Z; Burke M; Cogan D; Gao DA; Haverty K; Heim-Riether A; Hickey ER; Nagaraja R; Netherton M; O'Shea K; Ramsden P; Schwartz R; Shih D-T; Ward Y; Young E; Zhang Q
    Title:
    Exploration of cathepsin S inhibitors characterized by a triazole P1-P2 amide replacement.
    Source:
    Bioorg Med Chem Lett 22 (23), 7189-7193 (2012)
  • Author:
    Jenh C-H; Cox MA; Cui L; Reich E-P; Sullivan L; Chen S-C; Kinsley D; Qian S; Kim SH; Rosenblum S; Kozlowski J; Fine JS; Zavodny PJ; Lundell D
    Title:
    A selective and potent CXCR3 antagonist SCH 546738 attenuates the development of autoimmune diseases and delays graft rejection.
    Source:
    BMC Immunol 13 art.no.2 (2012)
    Abstract:
    Background: The CXCR3 receptor and its three interferon-inducible ligands (CXCL9, CXCL10 and CXCL11) have been implicated as playing a central role in directing a Th1 inflammatory response. Recent studies strongly support that the CXCR3 receptor is a very attractive therapeutic target for treating autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis and psoriasis, and to prevent transplant rejection. We describe here the in vitro and in vivo pharmacological characterizations of a novel and potent small molecule CXCR3 antagonist, SCH 546738.Results: In this study, we evaluated in vitro pharmacological properties of SCH 546738 by radioligand receptor binding and human activated T cell chemotaxis assays. In vivo efficacy of SCH 546738 was determined by mouse collagen-induced arthritis, rat and mouse experimental autoimmune encephalomyelitis, and rat cardiac transplantation models. We show that SCH 546738 binds to human CXCR3 with a high affinity of 0.4 nM. In addition, SCH 546738 displaces radiolabeled CXCL10 and CXCL11 from human CXCR3 with IC 50ranging from 0.8 to 2.2 nM in a non-competitive manner. SCH 546738 potently and specifically inhibits CXCR3-mediated chemotaxis in human activated T cells with IC 90about 10 nM. SCH 546738 attenuates the disease development in mouse collagen-induced arthritis model. SCH 546738 also significantly reduces disease severity in rat and mouse experimental autoimmune encephalomyelitis models. Furthermore, SCH 546738 alone achieves dose-dependent prolongation of rat cardiac allograft survival. Most significantly, SCH 546738 in combination with CsA supports permanent engraftment.Conclusions: SCH 546738 is a novel, potent and non-competitive small molecule CXCR3 antagonist. It is efficacious in multiple preclinical disease models. These results demonstrate that therapy with CXCR3 antagonists may serve as a new strategy for treatment of autoimmune diseases, including rheumatoid arthritis and multiple sclerosis, and to prevent transplant rejection.
  • Author:
    Uto-Konomi A; Miyauchi K; Ozaki N; Motomura Y; Suzuki Y; Yoshimura A; Suzuki S; Cua D; Kubo M
    Title:
    Dysregulation of suppressor of cytokine signaling 3 in keratinocytes causes skin inflammation mediated by interleukin-20 receptor-related cytokines
    Source:
    PLoS ONE 7 (7) art.no.e40343 (2012)
    Abstract:
    Homeostatic regulation of epidermal keratinocytes is controlled by the local cytokine milieu. However, a role for suppressor of cytokine signaling (SOCS), a negative feedback regulator of cytokine networks, in skin homeostasis remains unclear. Keratinocyte specific deletion of Socs3 (Socs3 cKO) caused severe skin inflammation with hyper-production of IgE, epidermal hyperplasia, and S100A8/9 expression, although Socs1 deletion caused no inflammation. The inflamed skin showed constitutive STAT3 activation and up-regulation of IL-6 and IL-20 receptor (IL-20R) related cytokines, IL-19, IL-20 and IL-24. Disease development was rescued by deletion of the Il6 gene, but not by the deletion of Il23, Il4r, or Rag1 genes. The expression of IL-6 in Socs3 cKO keratinocytes increased expression of IL-20R-related cytokines that further facilitated STAT3 hyperactivation, epidermal hyperplasia and neutrophilia. These results demonstrate that skin homeostasis is strictly regulated by the IL-6-STAT3-SOCS3 axis. Moreover, the SOCS3-mediated negative feedback loop in keratinocytes has a critical mechanistic role in the prevention of skin inflammation caused by hyperactivation of STAT3. © 2012 Uto-Konomi et al.
  • Author:
    Horvath C; Andrews L; Baumann A; Black L; Blanset D; Cavagnano J; Hastings K L; Hutto D L; MacLachlan T K; Milton M; Reynolds T; Robert S; Rogge M; Sims J; Treacy G; Warner G; Green J D
    Title:
    Storm forecasting: Additional lessons from the CD28 superagonist TGN1412 trial
    Source:
    Nat Rev Immunol 12 (10), 740 (2012)
    Abstract:
    no Abstract available
  • Author:
    Fu W; Ergun A; Lu T; Hill J A; Haxhinasto S; Fassett M S; Gazit R; Adoro S; Glimcher L; Chan S; Kastner P; Rossi D; Collins J J; Mathis D; Benoist C
    Title:
    A multiply redundant genetic switch 'locks in' the transcriptional signature of regulatory T cells
    Source:
    Nature Immunol 13 (10), 972-980 (2012)
    Abstract:
    The transcription factor Foxp3 participates dominantly in the specification and function of Foxp3 + CD4 + regulatory T cells (T reg cells) but is neither strictly necessary nor sufficient to determine the characteristic T reg cell signature. Here we used computational network inference and experimental testing to assess the contribution of other transcription factors to this. Enforced expression of Helios or Xbp1 elicited distinct signatures, but Eos, IRF4, Satb1, Lef1 and GATA-1 elicited exactly the same outcome, acting in synergy with Foxp3 to activate expression of most of the T reg cell signature, including key transcription factors, and enhancing occupancy by Foxp3 at its genomic targets. Conversely, the T reg cell signature was robust after inactivation of any single cofactor. A redundant genetic switch thus 'locked in' the T reg cell phenotype, a model that would account for several aspects of T reg cell physiology, differentiation and stability. © 2012 Nature America, Inc. All rights reserved.
  • Author:
    Wagner C L; Visvanathan S; Elashoff M; Mcinnes I B; Mease P J; Krueger G G; Murphy F T; Papp K; Gomez-Reino JJ; Mack M; Beutler A; Gladman D; Kavanaugh A
    Title:
    Markers of inflammation and bone remodelling associated with improvement in clinical response measures in psoriatic arthritis patients treated with golimumab
    Source:
    Ann Rheum Dis 72 (1), 83-88 (2013)
    Abstract:
    Objective: To determine serum biomarker associations with clinical response to golimumab treatment in patients with psoriatic arthritis (PsA). Methods: GO-REVEAL was a randomised, placebo-controlled study of golimumab in patients with active PsA. Samples were collected from 100 patients at baseline, week 4 and week 14, and analysed for serum-based biomarkers and protein profiling (total 92 markers); data were correlated with clinical measures at week 14. Results: Serum levels of a subset of proteins (apolipoprotein C III, ENRAGE, IL-16, myeloperoxidase, vascular endothelial growth factor, pyridinoline, matrix metalloproteinase 3, C-reactive protein (CRP), carcinoembryonic antigen, intercellular adhesion molecule 1 and macrophage inflammatory protein 1?) at baseline or week 4 were strongly associated with American College of Rheumatology 20% improvement (ACR20) response and/or disease activity score in 28 joints (DAS28) at week 14. A smaller subset of proteins was significantly associated with a 75% improvement in the psoriasis area and severity index score (PASI75) at week 14, (adiponectin, apolipoprotein CIII, serum glutamic oxaloacetic transaminase, and tumour necrosis factor ?). Subsets of proteins were identified as potentially predictive of clinical response for each of the clinical measures, and the power of these biomarker panels to predict clinical response to golimumab treatment was stronger than for CRP alone. Conclusions: This analysis provides insight into several panels of markers that may have utility in identifying PsA patients likely to have ACR20, DAS28, or PASI75 responses following golimumab treatment. Copyright Article author (or their employer) 2012.
  • Author:
    Horvath C; Andrews L; Baumann A; Black L; Blanset D; Cavagnaro J; Hastings K L; Hutto D L; MarLachlan T K; Milton M; Reynolds T; Roberts S; Roge M; Sims J; Treacy G; Warner G; Green J D
    Title:
    Storm forecasting: additional lessons from the CD28 superagonist TGN1412 trial
    Source:
    Nat Rev Immunol Article in Press (2012)
    Abstract:
    -
  • Author:
    Nakatsuji Y; Okuno T; Moriya M; Sugimoto T; Kinoshita M; Takamatsu H; Nojima S; Kimura T; Kang S; Ito D; Nakagawa Y; Toyofuku T; Takata K; Nakano M; Kubo M; Suzuki S; Matsui-Hasumi A; Uto-Konomi A; Ogata A; Mochizuki H; Sakoda S; Kumanogoh A
    Title:
    Elevation of Sema4A implicates Th cell skewing and the efficacy of IFN-beta therapy in multiple sclerosis
    Source:
    J Immunol 188 (10), 4858-4865 (2012)
    Abstract:
    Multiple sclerosis (MS) is a demyelinating autoimmune disease of the CNS and a leading cause of lasting neurologic disabilities in young adults. Although the precise mechanism remains incompletely understood, Ag presentation and subsequent myelin-reactive CD4 + T cell activation/differentiation are essential for the pathogenesis of MS. Although semaphorins were initially identified as axon guidance cues during neural development, several semaphorins are crucially involved in various phases of immune responses. Sema4A is one of the membrane-type class IV semaphorins, which we originally identified from the cDNA library of dendritic cell (DC). Sema4A plays critical roles in T cell activation and Th1 differentiation during the course of experimental autoimmune encephalomyelitis, an animal model of MS; however, its pathological involvement in human MS has not been determined. In this study, we report that Sema4A is increased in the sera of patients with MS. The expression of Sema4A is increased on DCs in MS patients and shed from these cells in a metalloproteinase- dependent manner. DC-derived Sema4A is not only critical for Th1 but also for Th17 cell differentiation, and MS patients with high Sema4A levels exhibit Th17 skewing. Furthermore, patients with high Sema4A levels have more severe disabilities and are unresponsive to IFN-b treatment. Taken together, our results suggest that Sema4A is involved in the pathogenesis of MS by promoting Th17 skewing. Copyright © 2012 by The American Association of Immunologists, Inc.
  • Author:
    Fu W; Ergun A; Lu T; Hill J A; Haxhinasto S; Fassett M S; Gazit R; Adoro S; Glimcher L; Chan S; Kastner P; Rossi D; Collins J J; Mathis D; Benoist C
    Title:
    A multiply redundant genetic switch 'locks in' the transcriptional signature of regulatory T cells
    Source:
    Nat Immun Article in Press (2012)
    Abstract:
    The transcription factor Foxp3 participates dominantly in the specification and function of Foxp3 +CD4 + regulatory T cells (T reg cells) but is neither strictly necessary nor sufficient to determine the characteristic T reg cell signature. Here we used computational network inference and experimental testing to assess the contribution of other transcription factors to this. Enforced expression of Helios or Xbp1 elicited distinct signatures, but Eos, IRF4, Satb1, Lef1 and GATA-1 elicited exactly the same outcome, acting in synergy with Foxp3 to activate expression of most of the T reg cell signature, including key transcription factors, and enhancing occupancy by Foxp3 at its genomic targets. Conversely, the T reg cell signature was robust after inactivation of any single cofactor. A redundant genetic switch thus 'locked in' the T reg cell phenotype, a model that would account for several aspects of T reg cell physiology, differentiation and stability.
  • Author:
    Dey AK; Burke B; Sun Y; Sirokman K; Nandi A; Hartog K; Lian Y; Geonnotti AR; Montefiori D; Franti M; Martin G; Carfi A; Kessler P; Martin L; Srivastava IK; Barnett SW
    Title:
    Elicitation of neutralizing antibidies directed against CD4-induced epitope(s) using a CD4 mimetic cross-linked to a HIV-1 envelope gllycoprotein.
    Source:
    PLoS ONE 7 (1) art. no. e30233 (2012)
    Abstract:
    The identification of HIV-1 envelope glycoprotein (Env) structures that can generate broadly neutralizing antibodies (BNAbs) is pivotal to the development of a successful vaccine against HIV-1 aimed at eliciting effective humoral immune responses. To that end, the production of novel Env structure(s) that might induce BNAbs by presentation of conserved epitopes, which are otherwise occluded, is critical. Here, we focus on a structure that stabilizes Env in a conformation representative of its primary (CD4) receptor-bound state, thereby exposing highly conserved "CD4 induced" (CD4i) epitope(s) known to be important for co-receptor binding and subsequent virus infection. A CD4-mimetic miniprotein, miniCD4 (M64U1-SH), was produced and covalently complexed to recombinant, trimeric gp140 envelope glycoprotein (gp140) using site-specific disulfide linkages. The resulting gp140-miniCD4 (gp140-S-S-M64U1) complex was recognized by CD4i antibodies and the HIV-1 co-receptor, CCR5. The gp140-miniCD4 complex elicited the highest titers of CD4i binding antibodies as well as enhanced neutralizing antibodies against Tier 1 viruses as compared to gp140 protein alone following immunization of rabbits. Neutralization against HIV-2 7312/V434M and additional serum mapping confirm the specific elicitation of antibodies directed to the CD4i epitope(s). These results demonstrate the utility of structure-based approach in improving immunogenic response against specific region, such as the CD4i epitope(s) here, and its potential role in vaccine application.
  • Author:
    Yi T; Fogal B; Hao Z; Tobiasova Z; Wang C; Rao DA; Al-Lamki RS; Kinkiles-Smith NC; Kulkami S; Bradley JR; Bothwell ALM; Sassa WC; Tellides G; Pober JS
    Title:
    Reperfusion injury intensifies the adaptive human T cell alloresponse in a human mouse chimeric artery model.
    Source:
    Arterioscler Thromb Vasc Biol 32 (2), 353-360 (2012)
  • Author:
    Gurtner K; Hessel F; Eicheler W; Doerfler A; Zips D; Heider K-H; Krause M; Baumann M
    Title:
    Combined treatment of the immunoconjgate bivatuzumab mertansine and fractionated irradiation improves local tumour control in vivo.
    Source:
    Radiother Oncol 102 (3), 444-449 (2012)
    Abstract:
    Background and purpose: To test whether BIWI 1 (bivatuzumab mertansine), an immunoconjugate of the humanized anti-CD44v6 monoclonal antibody BIWA 4 and the maytansinoid DM1, given simultaneously to fractionated irradiation improves local tumour control in vivo compared with irradiation alone. Material and methods: For growth delay, FaDu tumours were treated with 5 intravenous injections (daily) of phosphate buffered saline (PBS, control), BIWA 4 (monoclonal antibody against CD44v6) or BIWI 1 (bivatuzumab mertansine) at two different dose levels (50 ?g/kg DM1 and 100 ?g/kg DM1). For local tumour control, FaDu tumours received fractionated irradiation (5f/5d) with simultaneous PBS, BIWA 4 or BIWI 1 (two dose levels). Results: BIWI 1 significantly improved local tumour control after irradiation with 5 fractions already in the lower concentration. The dose modifying factor of 1.9 is substantial compared to the majority of other modifiers of radiation response. Conclusion: Because of the magnitude of the curative effect, this approach is highly promising and should be further evaluated using similar combinations with improved tumour-specificity.