bluuebunny/crossref_metadata_2025 huggingface dataset:
Consonant, Resonant and Social Relations between Firm and Consumer
Authors: Basile Gianpaolo
Published: April 2014

The aim of this chapter is to propose a conceptual framework, based on an interdisciplinary approach, which integrates the Viable Systems Perspective, Institutional and neo-Institutional Theory and Structuration Theory with the Consumer Behaviour Theory. The reason for this is to show the relational dynamics between the firm-brand and the individual-consumer, and between these and the direct-indirect stakeholder. The chapter is based on the reflection that the consumption system can be thought of as having two levels. The first is a micro-level, where the consumption system is characterized by the momentum of the creation and maintenance of consonant relationships between the firm-brand and the individual-consumer in order to achieve mutual systemic viability. The second is a macro-level, characterized by the concept that relationships between firm-brand and individual-consumer influence both direct and indirect stakeholders. Within these relationships, according to Stakeholder Theory, it is therefore very important to consider the reciprocal influences the indirect stakeholder has on and receives from the relationships. The results of these relationships affirm the legitimate conditions on which sustainability requirements (economic, social and environmental) are based. For these reasons the proposed conceptual framework will analyze the firm-brand three-dimensional social role when it is engaged to create and/or maintain and/or guarantee a long term enduring relationship.
Consumer Preferences and Key Aspects of Tourism and Hospitality Marketing on Island Destinations
Authors: Pappa Nikolaos
Published: April 2014

The decision of destination's selection is almost always accompanied by hotel selection. Hence, it is essential for tourism stakeholders to understand the marketing means' correlation toward destinations and hotels. In addition the purchasing behavior of consumers in tourism and hospitality needs to be examined in order to give the ability to optimize marketing strategies and promotional campaigns. The purpose of this chapter is to examine and present the perspectives on the island's destination and hospitality marketing held by consumers visiting Crete. The study's contribution is based on the clarification of consumers' purchase behavior and consumerism patters in tourism with reference to destination and hotel selection. It also gives an understanding for the marketing's influential importance for tourism purchases. The research was undertaken with departed tourists from Crete. As the research findings revealed, the perspectives of tourists concerning destination and hospitality marketing may vary significantly toward gender, age, education, and income. The research findings also indicate that younger and more educated consumers seem to have better knowledge and higher use of Information Technologies. On the contrary, elder people and those who are less educated are dependant to traditional ways of advertising. Furthermore, income plays a significant role dealing with additional services, better accommodation, and – in general – higher quality of provided products, whilst pricing policy still remains crucial for all tourists, no matter their financial revenues. The chapter also provides suggestions for further destination and hospitality marketing development, and information for the importance of each marketing mean used to lure tourists to visit the destination.
Consumerism and Innovation
Authors: Mosconi Enrico M. , Piccarozzi Michela , Silvestri Cecilia
Published: April 2014

In the current economic environment companies are progressively required to focus in innovation. In particular, consumerism is one of the major pulses to innovation. Today, consumers are looking for more and more innovative and qualitative products and so companies seek to heavily invest in technological advancements to meet the needs of customers. Even universities try to follow the trend of consumerism and adapt their activities to the market demands: first of all, in the way of teaching and transferring culture, and secondly, using their knowledge to create innovative companies. So, this second mission that allows universities to commercialize research results, has assumed a strategic role and primary importance in government policies, in Europe as well as in the rest of the world. University spin-off (USOs) can be considered one of the most important vehicle to create innovative firms based on scientific and new technological product. The trend of consumerism can be followed with spin-offs. The purpose of this chapter is to highlight the link between spin-off planning, especially during the business plan preparation, and consumerism. This area is investigated through a systematic review of the literature about the consumer, consumerism and the spin-off phenomenon.
Open Innovation through Customers
Authors: Aquilani Barbara , Abbate Tindara
Published: April 2014

This chapter aims at analyzing how firms can successfully embrace an Open Innovation (OI) process through customers, involving them, individually or in communities, in the co-creation of ideas, knowledge, products, services, processes, putting into action and integrating their creativity with firms' resources. Three main areas of interest are analyzed through a literature review process, to create a framework able to show the challenges organizations have to meet simultaneously externally (i.e. consumerism) and internally (i.e. organizational changes) in this shift of the innovation paradigm: consumerism features and challenges, OI approach and web-based platforms, and organizational issues involved in the OI paradigm shift. This chapter affords consumerism and OI approach, while the next, which is the sequel of this one, discusses OI platforms and organizational changes as well as the resulting framework. Four contributions distinguish this study: (i) the link between consumerism and OI; (ii) the focus on customers as a source of external innovation; (iii) the identification of alternative ways to access OI with customers and their features; (iv) the disclosure of a “hybrid” mode to develop OI through customers.
Open Innovation through Customers
Authors: Aquilani Barbara , Abbate Tindara
Published: April 2014

This chapter aims at analyzing how firms can successfully embrace an Open Innovation (OI) process through customers, involving them individually or in communities, in the co-creation of ideas, knowledge, products, services, processes, putting into action and integrating their creativity with firms' resources. Three main areas of interest, essential in the authors' opinion to meet this new challenge, have been analyzed through a literature review process, to create a framework able to show the challenges organizations have to meet externally (i.e., consumerism) and internally (i.e., organizational changes) in this shift of the innovation paradigm: consumerism features and challenges, OI approach and web-based platforms, and organizational issues involved in the OI paradigm shift. The previous chapter has discussed consumerism and OI approach, while this one will afford OI platforms and organizational changes as well as the resulting framework. Four contributions distinguish this study: (i) the link between consumerism and OI; (ii) the focus on customers as a source of external innovation; (iii) the identification of alternative ways to access OI with customers and their features; (iv) the disclosure of a “hybrid” mode to develop OI through customers.
Crowdsourcing Corporate Sustainability Strategies
Authors: Jones Peter , Hillier David , Comfort Daphne
Published: April 2014

This chapter provides cameo case studies of the way in which three large companies -- Unilever, Sony and J. Sainsbury -- have employed crowdsourcing exercises to review, refine and develop their corporate sustainability thinking, commitments and strategies. It also offers some wider reflections on this process. The chapter begins with an outline of the origins and basic characteristics of crowdsourcing, followed by a short discussion of the concept and contested meanings of sustainability. The material for the three cameo case studies is drawn from the reports Unilever, Sony and J. Sainsbury posted on the Internet about the crowdsourcing exercises they conducted to review their sustainability plans and strategies during 2010, 2011 and 2012. The findings reveal that the crowdsourcing exercises undertaken by the three companies attracted a wide range of constructive comments and produced a number of recommendations designed to develop new sustainability initiatives and to enhance their existing corporate sustainability strategies. While crowdsourcing offers a number of attractions to companies looking to review and develop sustainability strategies, not least in that it seems to offer a cost effective way of tapping into a number of other corporate organisations and to a potentially large number of consumers, it is not without its problems. The chapter provides an accessible exploratory review of how three major companies are employing crowdsourcing in developing and reviewing their corporate sustainability thinking, commitments and strategies.
Establishing Key Performance Indicators to Measure the Benefit of Introducing the Facilities Manager at an Early Stage in the Building Information Modeling Process
Authors: McAuley Barry , Hore Alan , West Roger
Published: April 2014

The Facilities Manager occupies a unique position within the lifecycle of a building asset, as he/she is one of the only Architecture/Engineering/Construction (AEC) and Facilities Management (FM) professionals who is in a position to view the product of the entire design and build process and is responsible for the operational phase which incurs approximately five times the initial capital cost. Despite the potential benefits that the Facilities Manager offers, this profession still does not command the recognition it deserves within the AEC/FM sector. This is beginning to change through the introduction of Building Information Modelling (BIM). In order for the BIM process to be maximised it is imperative that the Facilities Manager plays a much more important role within the design and construction process. Despite this, at present the role of the Facilities Manager within this process is still uncertain with no set Key Performance Indicators (KPI) or role designation being specified to date. This paper outlines how the Facilities Manager can play a pivotal role in the BIM process and, in particular, will aim to establish the basis for a number of KPIs by Facility Managers which will in turn lead to a more robust Lean FM practice.
A Theoretical Comparison of Traditional and Integrated Project Delivery Design Processes on International BIM Competitions
Authors: Serginson Michael , Mokhtar George , Kelly Graham
Published: April 2014

The Architectural Engineering and Construction (AEC) industry experiences higher rates of iteration, material wastage and poor cost management in comparison to other design industries. In an attempt to address such inefficiencies and control project budgets, various Governments are insisting that Building Information Modelling (BIM) is used by the appointed design teams on high value public buildings. Such legislation has been introduced in order to encourage a standardised level of collaborative working throughout the design process by enhancing interoperability of project information between design and construction professionals. In this paper, the MacLeamy Curve, a theoretical graphical representation of how integrated project delivery (IPD) processes improve efficiencies and allow for the reduction of costs by resolving issues during the earlier stages of the project, as well as other associated benefits are tested on both traditional and IPD design processes within two 48 hour international openBIM competition projects: Build London Live; and Build Qatar Live. The projects are compared by analysing the planned project programme against the reality, measured through recorded project exchanges, using a graphical representation. The findings of this paper suggest several recommendations, including: a collaborative design process appears to reduce iteration and results in a more comprehensive conceptual design at an early stage in comparison to a traditional process; more information and documentation is produced; and the overall programme is exceeded. Such findings suggest improved time, cost and design quality control.
Dialogue Journals and Transformational Learning
Authors: Nieves Yolanda
Published: April 2014

This article explores how dialogue journals can lead to a transformative learning experience. Adult Latino students enrolled in a community college developmental reading class agree to speak truth to power through this critical writing process. Using Mezirow's(2002) transformational learning theory, Brookfield's (2000) concepts on teaching for critical thinking, and Cranton's (2000) ideas of individuation and strategy for fostering self-awareness in students, the students and professor “talk-back” to each other through dialogue journals. The complexities of discourse, culture, and individuation or resistance to it are revealed.
Perceived Best Practices for Faculty Training in Distance Education
Authors: McVey Michael G.
Published: April 2014

Student learning style differences have been widely researched in both traditional face-to-face and online learning environments (Irani, Scherler & Harrington, 2003;Steinbronn, 2007; Williamson & Watson, 2007; Ugur, Akkoyunlu & Kurbanoglu, 2001). After conducting a literature review of adult student learning styles and teaching method analysis, it became apparent that there was not a significant difference in academic performance for students with differing learning styles whether they attended face-to-face or online classroom environments. What was not clearly indicated though, from the review of the literature, was what were the perceived best practices for online teaching from the perspective of experienced distance educators and whether the instructors' perceived learning style was incorporated in training programs to assist faculty to teach online. Thus, the purpose of this qualitative pilot study is to determine the perceived best practices to train faculty to teach in an online environment and how individual instructors' perceived learning style can be incorporated within best practices to foster competence on an individual instructor level. This study also analyzes faculty resistance to distance education and how transformative learning theory may play a role in overcoming this resistance.
Transformative Learning
Authors: Moss Leah , Brown Andy
Published: April 2014

Recognition of Acquired Competencies (RAC) as it is known in Quebec, Canada or Prior Learning Assessment (PLA), requires a learner to engage in retrospective thought about their learning path, their learning style and their experiential knowledge. This process of critical self-reflection and rigorous analysis by the learner of their prior learning is often the first exposure to the examination of their own knowledge. This article provides case studies of immigrant learners from the Montreal, Quebec area in a Recognition of Acquired Competencies process in vocational education. Through the analysis of interviews with learners, the authors suggest transformative learning is a by-product of the Recognition of Acquired Competencies process.
Towards a Decision Making Support System for the Capacitated Vehicle Routing Problem
Authors: Tlili Takwa , Krichen Saoussen , Faiz Sami
Published: April 2014

Distribution of goods is of importance in logistics and supply chain management. The target of a distribution network is to specify the beneficial way for delivering goods and commodities from supply to demand points. In distribution domain, this activity can usually be viewed as a capacitated vehicle routing problem (CVRP). As the CVRP solutions remain computationally intractable, we develop in this paper an interactive routing decision making support system (R-DMSS) that integrates a swarm based approach into a geographical information system (GIS). The empirical experiments include benchmarking instances as well as a case study over the Ezzahra area in Tunisia. The R-DMSS interfaces are presented to better understand the operational aspect of the system.
Extending QMBE Language with Clustering
Authors: Azevedo Ana , Santos Manuel Filipe
Published: April 2014

Business Intelligence (BI) is an important area of the Decision Support Systems (DSS) discipline. Over the past years, the evolution in this area has been considerable. Similarly, in the last years, there has been a huge growth and consolidation of the Data Mining (DM) field. DM is being used with success in BI systems, but a truly DM integration with BI is lacking. This creates a gap between DM and BI systems. With the purpose of closing this gap a new DM language for BI, named as Query-Models-By-Example (QMBE), was envisaged and implemented with success, but addressing only classification rules. This paper presents an extension of QMBE language to include clustering. This represents one more step towards the integration of DM with BI, which constitutes an important issue.
Relationship Conflicts Modeling and Measuring Within Work Teams
Authors: Wu Jiangning , Lei Fan
Published: April 2014

Nowadays many corporates make use of teams to complete tasks that individuals could not tackle alone. To effectively fulfill tasks, team members with different backgrounds work cooperatively and communicate constantly. In the process, relationship conflicts within the team inevitably occur due to clashes in personality and or values, which are empirically proved to be negative for completion of tasks. The paper focuses on the relationship conflicts resulted from exchanging and sharing knowledge between team members when completing the given tasks and accordingly develops a method to model and measure such conflicts. Based on the developed method, several simulations for the interaction of team members have been conducted to aim revealing the micro-mechanism of generation of relationship conflicts. Experimental results show that relationship conflicts can be generated and evolved with knowledge transfer between team members. Furthermore, impact factors on relationship conflicts as well as team creativity are deeply analyzed from the personality point of view.
Linguistic Multi-Attribute Decision Making with a Prioritization Relationship
Authors: Wei Cuiping , Tang Xijin , Wang Xiaojie
Published: April 2014

In this paper we consider linguistic information aggregation problems where a prioritization relationship exists over attributes. The authors define a prioritized 2-tuple ordered weighted averaging (PTOWA) operator to aggregate satisfactions of alternatives under attributes with a linear prioritized ordering. The authors then use the PTOWA and a TOWA operator to aggregate linguistic information where attributes are partitioned into some categories of which prioritization between categories exists. Finally, two illustrative examples are employed to show the feasibility of the proposed method.
IFN-γ-Inducible T Cell α Chemoattractant Is a Potent Stimulator of Normal Human Blood T Lymphocyte Transendothelial Migration: Differential Regulation by IFN-γ and TNF-α
Authors: Mohan Karkada , Ding Ziqiang , Hanly John , Issekutz Thomas B.
Published: April 2014

Previous studies have shown that the CXC chemokine, IFN-γ-inducible T cell α chemoattractant (I-TAC), was chemotactic for IL-2-activated human T lymphocytes, which express abundant CXCR3. However, because most memory T lymphocytes are also CXCR3+, the ability of I-TAC to promote the migration of normal human blood T cells across HUVEC monolayers in Transwell chambers was examined. I-TAC induced a marked (4- to 6-fold) increase in transendothelial migration (TEM) of T cells across unstimulated HUVEC from 5.6 to 28% of input T cells and was substantially more active than IFN-γ-inducible protein-10, another CXCR3 ligand. I-TAC significantly enhanced TEM of T cells across TNF-α, but not across IFN-γ or IFN-γ plus TNF-α-activated HUVEC. IFN-γ or IFN-γ plus TNF-α-activated HUVEC produced substantial amounts of I-TAC, in contrast to TNF-α-treated EC. Both CD4+ and CD8+ T cells migrated in response to I-TAC to a similar extent, while memory T cells migrated several fold better than naive T cells. Blockade of LFA-1 strongly inhibited I-TAC-induced T cell TEM across unstimulated HUVEC, and ∼50–60% of the TEM across cytokine-activated HUVEC. However, blocking both LFA-1 and very late Ag-4 abolished I-TAC induced T cell TEM. In vivo significant levels of I-TAC were detected in arthritic synovial fluid. Thus, I-TAC is one of the most potent chemoattractants of normal human blood CD4 and CD8 T cell TEM and is likely a major mediator of blood memory T lymphocyte migration to inflammation.
Transgenic Expression of a Human Polyreactive Ig Expressed in Chronic Lymphocytic Leukemia Generates Memory-Type B Cells That Respond to Nonspecific Immune Activation
Authors: Widhopf George F. , Brinson Diana C. , Kipps Thomas J. , Tighe Helen
Published: April 2014

We generated transgenic mice, designated SMI, expressing unmutated H and L chain Ig genes encoding a low-affinity, polyreactive human (h)IgM/κ rheumatoid factor. These animals were compared with control AB29 transgenic mice expressing a hIgM/κ rheumatoid factor specific for human IgG, with no detectable reactivity with mouse proteins. SMI B cells expressed significantly lower levels of surface hIgM/κ than did the B cells of AB29 mice, but still could be induced to proliferate by surface Ig cross-linking in vitro and could be deleted with anti-Id mAb in vivo. Transgene-expressing B cells of AB29 mice had a B-2 phenotype and were located in the primary follicle. In contrast, a relatively high proportion of hIgM-expressing B cells of SMI mice had the phenotype of B-1 B cells in the peritoneum or marginal zone B cells in the spleen, where they were located in the periarteriolar sheath, marginal zone, and interfollicular areas that typically are populated by memory-type B cells. Although the relative proportions of transgene-expressing B cells in both types of transgenic mice declined with aging, SMI mice experienced progressive increases in the serum levels of IgM transgene protein over time. Finally, SMI transgene-expressing B cells, but not AB29 transgene-expressing B cells, were induced to secrete Ab when cultured with alloreactive T cells. These results indicate that expression of polyreactive autoantibodies can allow for development of B cells that are neither deleted nor rendered anergic, but instead have a phenotype of memory-type or Ag-experienced B cells that respond to nonspecific immune activation.
Distinct Expression Profiles of the Peripheral Cannabinoid Receptor in Lymphoid Tissues Depending on Receptor Activation Status
Authors: Rayman Nazik , Lam King H. , Laman Jon D. , Simons Peter J. , Löwenberg Bob , Sonneveld Pieter , Delwel Ruud
Published: April 2014

Using two distinct anti-CB2 receptor Abs, we investigated the expression patterns of the peripheral cannabinoid receptor CB2 in human secondary lymphoid organs. Immunohistochemical analysis using an N-terminal specific anti-CB2 Ab revealed high protein expression in the germinal centers (GCs) of secondary follicles. A C-terminal specific anti-CB2 Ab, which only recognizes a nonphosphorylated inactive receptor, showed positivity in the mantle zones (MZs) and marginal zones (MGZs) of the secondary follicles where resting cells reside, and in the primary follicles. In contrast, no positivity was observed in GCs using the C-terminal Ab, suggesting that active CB2 receptors are mainly present on cells in the GCs. Dual immunohistochemical analysis revealed that B lymphocytes express the CB2 protein abundantly. In contrast to B cells in the MZ or MGZ, CB2-expressing cells in the GCs coexpress the costimulatory membrane protein CD40, which is mainly expressed in the GCs and at very low levels in the MZs and MGZs and the proliferation marker Ki-67. Using the human Raji B cell line as a model, we demonstrate in a transwell assay that moderate migration occurs upon stimulation of the CB2 receptor with the endocannabinoid 2-arachidonoylglycerol, which is enhanced by CD40 costimulation. Our findings, that GC-related cells express active CB2 and that CB2-dependent migration requires CD40 costimulation, suggest that CB2 is involved in B cell activation.
Molecular Characterization of Human 4Ig-B7-H3, a Member of the B7 Family with Four Ig-Like Domains
Authors: Steinberger Peter , Majdic Otto , Derdak Sophia V. , Pfistershammer Katharina , Kirchberger Stefanie , Klauser Christoph , Zlabinger Gerhard , Pickl Winfried F. , Stöckl Johannes , Knapp Walter
Published: April 2014

In an effort to characterize molecules with immunoregulatory potential, we raised mAbs to human dendritic cells. We selected an Ab that recognizes a molecule that is induced on monocytes differentiated in vitro toward dendritic cells. Retroviral expression cloning identified this molecule as B7-H3, a member of the B7 family described recently. In contrast to an earlier report, in which B7-H3 was described as a molecule consisting of two Ig-like domains, our cDNA encoded a type I membrane protein with four Ig-like domains, and the molecule identified by us was therefore named 4Ig-B7-H3. mRNA analysis as well as Western blotting experiments performed by us did not reveal evidence for a small B7-H3. B7-H3 is not expressed on peripheral blood lymphocytes, monocytes, or granulocytes. Upon in vitro stimulation, the expression of B7-H3 is induced on T cells, B cells, and NK cells. A number of different approaches were used to investigate the function of human B7-H3. In contrast to an earlier report, our data do not support a costimulatory role of B7-H3 in anti-CD3-mediated activation of the TCR-complex resulting in T cell proliferation and IFN-γ production.
Construction of Hevein (Hev b 6.02) with Reduced Allergenicity for Immunotherapy of Latex Allergy by Comutation of Six Amino Acid Residues on the Conformational IgE Epitopes
Authors: Karisola Piia , Mikkola Jari , Kalkkinen Nisse , Airenne Kari J. , Laitinen Olli H. , Repo Susanna , Pentikäinen Olli T. , Reunala Timo , Turjanmaa Kristiina , Johnson Mark S. , Palosuo Timo , Kulomaa Markku S. , Alenius Harri
Published: April 2014

Recently we have established that IgE Abs bind to conformational epitopes in the N- and C-terminal regions of the major natural rubber latex allergen, hevein (Hev b 6.02). To identify the critical amino acid residues that interact with IgE, the hevein sequence was scanned by using site-specific mutations. Twenty-nine hevein mutants were designed and produced by a baculovirus expression system in insect cells and tested by IgE inhibition-ELISA using sera from 26 latex allergic patients. Six potential IgE-interacting residues of hevein (Arg5, Lys10, Glu29, Tyr30, His35, and Gln38) were identified and characterized further in detail. Based on these six residues, two triple mutants (HΔ3A, HΔ3B) and hevein mutant where all six residues were mutated (HΔ6), were designed, modeled, and produced. Structural and functional properties of these combinatory mutants were compared experimentally and in silico with those of recombinant hevein. The IgE-binding affinity of the mutants decreased by three to five orders of magnitude as compared with that of recombinant hevein. Skin prick test reactivity of the triple mutant HΔ3A was drastically reduced and that of the six-residue mutant HΔ6 was completely abolished in all patients examined in this study. The approach presented in this paper offers tools for identification and modification of amino acid residues on conformational epitopes of allergens that interact with IgE. Hevein with a highly reduced ability to bind IgE should provide a valuable candidate molecule for immunotherapy of latex allergy and is anticipated to have a low risk of systemic side effects.
A Synthetic Peptide Homologous to Functional Domain of Human IL-10 Down-Regulates Expression of MHC Class I and Transporter Associated with Antigen Processing 1/2 in Human Melanoma Cells
Authors: Kurte Mónica , López Mercedes , Aguirre Adam , Escobar Alejandro , Aguillón Juan Carlos , Charo Jehad , Larsen Christian G. , Kiessling Rolf , Salazar-Onfray Flavio
Published: April 2014

Tumor cells treated with IL-10 were shown to have decreased, but peptide-inducible expression of MHC class I, decreased sensitivity to MHC class I-restricted CTL, and increased NK sensitivity. These findings could be explained, at least partially, by a down-regulation of TAP1/TAP2 expression. In this study, IT9302, a nanomeric peptide (AYMTMKIRN), homologous to the C-terminal of the human IL-10 sequence, was demonstrated to mimic these previously described IL-10 effects on MHC class I-related molecules and functions. We observed a dose-dependent down-regulation of MHC class I at the cell surface of melanoma cells after 24-h treatment with IT9302. The IL-10 homologue peptide also caused a dose-dependent inhibition of the IFN-γ-mediated surface induction of MHC class I in a melanoma cell line. We demonstrated, using Western blot and flow cytometry, that IT9302 inhibits the expression of TAP1 and TAP2 proteins, but not MHC class I H chain or low molecular protein molecules. Finally, peptide-treated melanoma cells were shown to be more sensitive to lysis by NK cells in a dose-dependent way. Taken together, these results demonstrate that a small synthetic peptide derived from IL-10 can mimic the Ag presentation-related effects mediated by this cytokine in human melanomas and increase tumor sensitivity to NK cells, which can be relevant in the designing of future strategies for cancer immune therapy.
Mouse Lysozyme-M Knockout Mice Reveal How the Self-Determinant Hierarchy Shapes the T Cell Repertoire against This Circulating Self Antigen in Wild-Type Mice
Authors: Sinha Pratima , Chi Howard H. , Kim Hong R. , Clausen Björn E. , Pederson Brian , Sercarz Eli E. , Forster Irmgard , Moudgil Kamal D.
Published: April 2014

We have studied T cell tolerance to defined determinants within ML-M using wild-type (WT; ML-M+/+) and LysMcre (ML-M−/−) C3H (H-2k) mice to determine the relative contribution of ML-M-derived epitopes vs those from other self Ags in selection of the ML-M-specific T cell repertoire. ML-M was totally nonimmunogenic in WT mice, but was rendered immunogenic in LysMcre mice. Most of the response to ML-M in LysMcre mice was directed to the immunodominant determinant region 105–119. This determinant is spontaneously displayed (without adding exogenous ML-M) by macrophages of WT, but not LysMcre, mice and is stimulatory for peptide 105–119 (p105–119)-primed T cells. Moreover, neonatal tolerization of LysMcre mice with p105–119 or ML-M abrogated the T cell response to subsequent challenge with ML-M or p105–119. Furthermore, p95–109 and p110–125 of ML-M were immunogenic in LysMcre mice, but not in WT mice, thereby representing subdominant, tolerance-inducing epitopes of ML-M. As expected, the T cell repertoire to cryptic ML determinants in WT mice was also intact in LysMcre mice. Furthermore, the pattern of response to the related homologue of ML-M, hen eggwhite lysozyme, was similar in these two groups of mice. Thus, several codominant T cell determinants within ML-M contribute significantly to tolerance induction, and the anti-cryptic T cell repertoire to ML-M was positively selected on non-ML-M self ligands. These results reveal that the induction of self tolerance to a multideterminant protein follows the quantitative hierarchy of self-determinant expression and are of relevance in understanding the pathogenesis of autoimmunity.
Endogenous Expression of Activation-Induced Cytidine Deaminase in Cell Line WEHI-231
Authors: Spillmann Freia J. X. , Wabl Matthias
Published: April 2014

Because of its susceptibility to apoptosis on Ag receptor cross-linking, cells of the mouse cell line WEHI-231 have been classified as immature B cells. Surprisingly, however, the cell line expresses activation-induced cytidine deaminase, the enzyme that mediates hypermutation and Ig class switch recombination in activated B cells. Although both cDNA sequence and protein expression of activation-induced cytidine deaminase appear normal, the cell line does not hypermutate an indicator plasmid. For the readout, the indicator plasmid depends on the removal of deoxyuridine after transition from C to U and, therefore, on functional expression of uracil N-glycosylase 2, which is normal in WEHI-231. At the endogenous Ig locus, however, WEHI-231 does undergo the canonical hypermutation of G · C to A · T base pairs to some extent. The cell line also expresses the germline transcripts of the Ig γ2b, ε, and α loci, but it does not switch its IgM surface Ig.
Blockade of CXCR3 Receptor:Ligand Interactions Reduces Leukocyte Recruitment to the Lung and the Severity of Experimental Idiopathic Pneumonia Syndrome
Authors: Hildebrandt Gerhard C. , Corrion Leigh A. , Olkiewicz Krystyna M. , Lu Bao , Lowler Kathleen , Duffner Ulrich A. , Moore Bethany B. , Kuziel William A. , Liu Chen , Cooke Kenneth R.
Published: April 2014

Idiopathic pneumonia syndrome (IPS) is a frequently fatal complication after allogeneic stem cell transplantation (allo-SCT) that responds poorly to standard immunosuppressive therapy. The pathophysiology of IPS involves the secretion of inflammatory cytokines including IFN-γ and TNF-α along with the recruitment of donor T cells to the lung. CXCR3 is a chemokine receptor that is expressed on activated Th1/Tc1 T cell subsets and the expression of its ligands CXCL9 (monokine induced by IFN-γ (Mig)) and CXCL10 (IFN-γ-inducible protein 10 (IP-10)) can be induced in a variety of cell types by IFN-γ alone or in combination with TNF-α. We used a lethally irradiated murine SCT model (B6 → bm1) to evaluate the role of CXCR3 receptor:ligand interactions in the development of IPS. We found that Mig and IP-10 protein levels were significantly elevated in the bronchoalveolar lavage fluid of allo-SCT recipients compared with syngeneic controls and correlated with the infiltration of IFN-γ-secreting CXCR3+ donor T cells into the lung. The in vivo neutralization of either Mig or IP-10 significantly reduced the severity of IPS compared with control-treated animals, and an additive effect was observed when both ligands were blocked simultaneously. Complementary experiments using CXCR3−/− mice as SCT donors also resulted in a significant decrease in IPS. These data demonstrate that interactions involving CXCR3 and its primary ligands Mig and IP-10 significantly contribute to donor T cell recruitment to the lung after allo-SCT. Therefore, approaches focusing on the abrogation of these interactions may prove successful in preventing or treating lung injury that occurs in this setting.
A Stable Aspirin-Triggered Lipoxin A4 Analog Blocks Phosphorylation of Leukocyte-Specific Protein 1 in Human Neutrophils
Authors: Ohira Taisuke , Bannenberg Gerard , Arita Makoto , Takahashi Minoru , Ge Qingyuan , Van Dyke Thomas E. , Stahl Gregory L. , Serhan Charles N. , Badwey John A.
Published: April 2014

Lipoxins and their aspirin-triggered 15-epimers are endogenous anti-inflammatory agents that block neutrophil chemotaxis in vitro and inhibit neutrophil influx in several models of acute inflammation. In this study, we examined the effects of 15-epi-16-(p-fluoro)-phenoxy-lipoxin A4 methyl ester, an aspirin-triggered lipoxin A4-stable analog (ATLa), on the protein phosphorylation pattern of human neutrophils. Neutrophils stimulated with the chemoattractant fMLP were found to exhibit intense phosphorylation of a 55-kDa protein that was blocked by ATLa (10–50 nM). This 55-kDa protein was identified as leukocyte-specific protein 1, a downstream component of the p38-MAPK cascade in neutrophils, by mass spectrometry, Western blotting, and immunoprecipitation experiments. ATLa (50 nM) also reduced phosphorylation/activation of several components of the p38-MAPK pathway in these cells (MAPK kinase 3/MAPK kinase 6, p38-MAPK, MAPK-activated protein kinase-2). These results indicate that ATLa exerts its anti-inflammatory effects, at least in part, by blocking activation of the p38-MAPK cascade in neutrophils, which is known to promote chemotaxis and other proinflammatory responses by these cells.
Protective Role of Arginase in a Mouse Model of Colitis
Authors: Gobert Alain P. , Cheng Yulan , Akhtar Mahmood , Mersey Benjamin D. , Blumberg Darren R. , Cross Raymond K. , Chaturvedi Rupesh , Drachenberg Cinthia B. , Boucher Jean-Luc , Hacker Amy , Casero Robert A. , Wilson Keith T.
Published: April 2014

Arginase is the endogenous inhibitor of inducible NO synthase (iNOS), because both enzymes use the same substrate, l-arginase (Arg). Importantly, arginase synthesizes ornithine, which is metabolized by the enzyme ornithine decarboxylase (ODC) to produce polyamines. We investigated the role of these enzymes in the Citrobacter rodentium model of colitis. Arginase I, iNOS, and ODC were induced in the colon during the infection, while arginase II was not up-regulated. l-Arg supplementation of wild-type mice or iNOS deletion significantly improved colitis, and l-Arg treatment of iNOS−/− mice led to an additive improvement. There was a significant induction of IFN-γ, IL-1, and TNF-α mRNA expression in colitis tissues that was markedly attenuated with l-Arg treatment or iNOS deletion. Treatment with the arginase inhibitor S-(2-boronoethyl)-l-cysteine worsened colitis in both wild-type and iNOS−/− mice. Polyamine levels were increased in colitis tissues, and were further increased by l-Arg. In addition, in vivo inhibition of ODC with α-difluoromethylornithine also exacerbated the colitis. Taken together, these data indicate that arginase is protective in C. rodentium colitis by enhancing the generation of polyamines in addition to competitive inhibition of iNOS. Modulation of the balance of iNOS and arginase, and of the arginase-ODC metabolic pathway may represent a new strategy for regulating intestinal inflammation.
Central Memory CD4+ T Cell Responses in Chronic HIV Infection Are Not Restored by Antiretroviral Therapy
Authors: Elrefaei Mohamed , McElroy Michael D. , Preas Christopher P. , Hoh Rebecca , Deeks Steven , Martin Jeffrey , Cao Huyen
Published: April 2014

A strong CD4+ T cell response has been correlated with better control of HIV infection. However, the effect of HIV on the maintenance of Ag-specific memory CD4+ T cells is not fully understood. We characterized the function and phenotype of memory CD4+ T cells generated by mumps and influenza A or B viruses in HIV-infected individuals receiving highly active antiretroviral therapy (n = 21), HIV-infected long-term nonprogressors (n = 10), and HIV-seronegative volunteers (n = 10). We observed significantly decreased proliferation of the Ag-specific central memory CD4+ T cell population (CD28+/CCR7+/CD45RA−) in the antiretroviral treated HIV-infected individuals compared with the seronegative controls. Restored CD4+ T cell count and decreased HIV viral load while on highly active antiretroviral therapy did not result in increased proliferation, whereas nadir CD4+ T cell count predicted the presence of Ag-specific proliferation. Our results indicate that HIV infection leads to impaired maintenance of virus-induced or vaccine-generated central memory CD4+ T cells that is not restored by HAART.
Induction of Human T Cell Leukemia Virus Type I Receptors on Quiescent Naive T Lymphocytes by TGF-β
Authors: Jones Kathryn S. , Akel Salem , Petrow-Sadowski Cari , Huang Ying , Bertolette Daniel C. , Ruscetti Francis W.
Published: April 2014

The retrovirus human T cell leukemia virus (HTLV) type I (HTLV-I) is primarily transmitted by breast-feeding or sexual contact, by cell-to-cell contact between T cells. TGF-β, which has been shown to enhance transmission of HTLV-I in vitro, is found at high levels in breast milk and semen. In this study, the ability of TGF-β to regulate expression of molecules involved in HTLV-I binding and entry was examined. Previous studies using a soluble form of the HTLV-I envelope protein SU have shown that quiescent human T cells do not express cell surface molecules that specifically bind SU. After T cell activation, HTLV SU binding proteins are rapidly induced. In this study, we report that TGF-β induces expression of proteins that bind soluble HTLV SU and HTLV virions on naive CD4+ T lymphocytes. The induction of these proteins occurred without cell cycle entry or expression of activation markers, involved TGF-β-induced intracellular signaling, and required de novo transcription and translation. Treatment of naive CD4+ T lymphocytes with TGF-β induced expression of GLUT-1, which has recently been reported to function as a receptor for HTLV. Treatment of a TGF-β-sensitive human myeloid cell line increased the titer of both HTLV-I- and HTLV-II-pseudotyped viruses. Although earlier studies suggested that HTLV SU binding proteins might be an early marker of T cell activation and/or cell proliferation, we report in this study that TGF-β induces binding of HTLV virions and expression of glucose transporter type 1 in primary CD4+ T lymphocytes that remain quiescent.
Identification of a Macrophage-Specific Chromatin Signature in the IL-10 Locus
Authors: Saraiva Margarida , Christensen Jillian R. , Tsytsykova Alla V. , Goldfeld Anne E. , Ley Steven C. , Kioussis Dimitris , O′Garra Anne
Published: April 2014

The molecular mechanisms that regulate expression of the immunosuppressive cytokine IL-10 remain poorly understood. In this study, by measuring sensitivity to DNase I digestion, we show that production of IL-10 by primary mouse bone marrow-derived macrophages stimulated through pattern recognition receptors was associated with chromatin remodeling of the IL-10 locus. We also demonstrate that the IL-10 locus is remodeled in primary Th2 cells and IL-10-producing regulatory T cells that have been differentiated in vitro. Strikingly, a novel DNase I-hypersensitive site (HSS-4.5) was identified in stimulated macrophages, but not in T cells. We show that hyperacetylated histones were recruited to this site in stimulated macrophages. Furthermore, HSS-4.5 is highly conserved and contains a putative NF-κB binding site. In support of a function for this site, NF-κB p65/RelA was recruited to HSS-4.5 in vivo and its activation was required for optimal IL-10 gene expression in LPS-stimulated macrophages.
A Critical Role for Sialylation in Cryoglobulin Activity of Murine IgG3 Monoclonal Antibodies
Authors: Kuroda Yasuhiro , Kuroki Aki , Kikuchi Shuichi , Funase Takaaki , Nakata Munehiro , Izui Shozo
Published: April 2014

Cryoprecipitating IgG3 autoantibodies have been shown to play a significant role in the development of murine lupus-like autoimmune syndrome. However, the structural basis of IgG3 cryoprecipitation still remains to be defined. In view of the implication of positively charged amino acid residues present in variable regions in IgG3 cryoglobulin activity, we explored the role of terminal sialic acids in oligosaccharide side chains for the cryogenic activity of IgG3 mAb. Comparative oligosaccharide structural analysis of different cryogenic and non-cryogenic IgG3 mAb showed an inverse correlation between the extent of sialylation and cryogenic activity. The inhibitory role of sialylation was further confirmed by the demonstration of enrichment of less and more sialylated IgG3 in cryoprecipitated and noncryoprecipitated fractions, respectively, separated from four different cryogenic IgG3 mAb. Significantly, the sialic acid contents of the latter fraction became comparable to those of non-cryogenic IgG3 mAb. Finally, we observed that highly sialylated non-cryogenic IgG3 mAb was more potent in the inhibition of cryoprecipitation of cryogenic IgG3 mAb. Our results thus suggest that the content of negatively charged sialic acids in oligosaccharide side chains is one of the critical factors to determine IgG3 cryoglobulin activity, along with amino acid sequences of the IgG3 variable regions.
Influence of β1 Integrin Intracytoplasmic Domains in the Regulation of VLA-4-Mediated Adhesion of Human T Cells to VCAM-1 under Flow Conditions
Authors: Rosenthal-Allieri Maria Alessandra , Ticchioni Michel , Breittmayer Jean Philippe , Shimizu Yoji , Bernard Alain
Published: April 2014

The VLA-4 integrin supports static cell-cell, cell-matrix adhesion, and dynamic interactions with VCAM-1. Although functions for well-conserved β1 integrin cytoplasmic domains in regulating static cell adhesion has been established, the molecular basis for β1 integrin-dependent arrest on VCAM-1 under flow conditions remains poorly understood. We have transfected the β1 integrin-deficient A1 Jurkat T cell line with β1 cDNA constructs with deletions of the NPXY motifs and specific mutations of tyrosine residues. Deletion of either NPXY motif impaired static adhesion induced by CD2 or CD47 triggering or direct β1 integrin stimulation. In contrast, PMA-induced adhesion to VCAM-1 was unaffected by deletion of the NPIY motif and only slightly impaired by deletion of NPKY. Moreover, deletion of the NPIY motif resulted in enhanced rolling and reduced arrest on VCAM-1 under shear flow conditions. In contrast, deletion of the NPKY motif did not alter arrest under flow. Although tyrosine to phenylalanine substitutions within two NPXY motifs did not alter static adhesion to VCAM-1, these mutations enhanced arrest on VCAM-1 under flow conditions. Furthermore, although deletion of the C′-terminal 5 AA of the β1 cytoplasmic domain dramatically impaired activation-dependent static adhesion, it did not impair arrest on VCAM-1 under flow conditions. Thus, our results demonstrate distinct structural requirements for VLA-4 function under static and shear flow conditions. This may be relevant for VLA-4 activity regulation in different anatomic compartments, such as when circulating cells arrest on inflamed endothelium under shear flow and when resident cells in bone marrow interact with VCAM-1- positive stromal cells.
Genetic Dissection of Systemic Lupus Erythematosus Pathogenesis: Partial Functional Complementation betweenSle1andSle3/5Demonstrates Requirement for Intracellular Coexpression for Full Phenotypic Expression of Lupus
Authors: Wakui Masatoshi , Morel Laurence , Butfiloski Edward J. , Kim Chunsun , Sobel Eric S.
Published: April 2014

Sle1 on chromosome 1 and Sle3/5 on chromosome 7 are two of the most critical lupus susceptibility loci of the New Zealand Black/White-derived NZM2410 mouse strain. In contrast to C57BL/6 mice congenic for either Sle1 (B6.Sle1) or Sle3/5 (B6.Sle3/5), strains that express only a modest lupus-related phenotype, the bicongenic B6.Sle1.Sle3/5 strain has a robust phenotype, suggesting a critical role for epistatic interactions in lupus pathogenesis. Mixed chimera experiments indicated that the two loci are functionally expressed by different cell populations and predicted that phenotypic expression of the phenotypic features of the B6.Sle1.Sle3/5 strain could be fully reproduced with a combination of B6.Sle1 and B6.Sle3/5 bone marrow. Contrary to our expectations, there was only a partial functional complementation in these mixed chimeras. Spleen enlargement, CD4:CD8 ratio elevation, and epitope spreading of autoantibodies were fully developed in B6+B6.Sle1.Sle3/5 but not in B6.Sle1+B6.Sle3/5 mixed chimeras. This study is the first to present evidence that the pathways mediated by two critical lupus susceptibility loci derived from the New Zealand White strain must be integrated intracellularly for epistatic interactions to occur. Our mixed chimera approach continues to provide novel insights into the functional genetic pathways underlying this important murine model of systemic autoimmunity.