266 research outputs found
Endogenously produced urokinase amplifies tumor necrosis factorâα secretion by THPâ1 mononuclear phagocytes
This study examined the effects of endogenous urokinase (uPA) on lipopolysaccharide (LPS)âstimulated tumor necrosis factor α (TNFâα) secretion in THPâ1 mononuclear phagocytes. AntiâuPA monoclonal antibody (mAb) suppressed LPSâdriven TNFâα secretion by 61.6 ± 5.9% (P < .001), and PAIâ1, a uPA inhibitor, suppressed it to 53.1 ± 8.2% of the control value (P < .001). Upâregulation of TNFâα mRNA was suppressed in parallel with secreted TNFâα protein. TNFâα secretion was unaffected by depleting plasminogen or by aprotinin, a plasmin inhibitor. When endogenous uPA was displaced from the cell, exogenous highâmolecularâweight (intact) uPA augmented LPSâdriven TNFâα secretion. By contrast, a uPA fragment containing the catalytic domain was inhibitory, and the uPA receptorâbinding domain had no effect. We conclude that endogenous uPA amplifies TNFâα neosynthesis of UPSâstimulated THPâ1 mononuclear phagocytes. The effect requires intact uPA and is independent of plasmin activity. This represents a novel mechanism by which a mononuclear phagocyteâderived protease contributes to generating proinflammatory signals.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/142208/1/jlb0302.pd
Formulation development of a recombinant protein based non-replicating rotavirus (NRRV) vaccine candidate: Antigen-adjuvant-preservative interactions
Rotavirus is the leading cause of acute diarrhea and gastroenteritis among infants and young children worldwide. Over 215,000 children under five years of age die from rotavirus infection each year, mostly in developing world1. Currently two live attenuated oral rotavirus vaccines are available globally (RotarixÂź and RotaTeqÂź) to reduce the burden of this disease with an efficacy of \u3e90% in developed countries2. Vaccine efficacy is lower, however, in developing countries due to a variety of factors3. To this end, a non-replicating rotavirus (NRRV) vaccine candidate, containing three recombinant protein antigens (P2-VP8-P[4], P2-VP8-P[6] and P2-VP8-P[8]), is being developed by PATH and its partners as a trivalent vaccine for use in the developing world4. This trivalent rotavirus vaccine candidate includes the three antigens from the most prevalent serotypes associated with \u3e90% of rotavirus gastroenteritis worldwide.
In the present study, the following formulation development issues were examined: (1) establish stability-indicating physicochemical assays for a NRRV protein antigen (P[8]) bound to an aluminum hydroxide adjuvant (AlhydrogelÂź), which include primary and higher-order structures, chemical and conformational stability of the protein on Alhydrogel, and the ability to desorb the antigen from Alhydrogel; (2) examine the adsorptive capacity and coefficients of AlhydrogelÂź for the P[8] antigen in several candidate drug product formulations; (3) investigate the effects of binding to AlhydrogelÂź and the addition of two antimicrobial preservatives (2-phenoxyethanol or thimerosal) on the structural integrity and conformational stability of P[8], the latter of which were found to be potent destabilizers of the antigen; and (4) monitor the real-time and accelerated storage stability over 3 months of P[8] bound to AlhydrogelÂź in several candidate formulations with and without thimerosal at different temperatures. In the absence of preservative, the P[8] protein antigen was overall stable with only a small amount of Asn deamidation observed in samples stored under real-time (4ËC) or accelerated (25ËC) temperatures. Similarly, little (if any) changes were observed in the real-time stability of the antigen on AlhydrogelÂź in the presence of thimerosal. Under accelerated storage temperatures (25 or 37ËC) however, the preservative caused an increase in inter-molecular disulfide bonding, decrease of apparent enthalpy as measured by DSC, and a decrease in in-vitro antigenicity. Similar stability studies are currently ongoing with the P[4] and P[6] protein antigens.
Acknowledgements: Funding provided by the Bill & Melinda Gates Foundation
References:
1. Tate et al 2016. Clinical Infectious Diseases 62:S96-S105
2. Tissera et al. 2017. Human Vaccines & Immunotherapeutics 13(4):921-927
3. Glass et al. 2014. Journal of Infection 68: S9-S18.
4. Groome et al. 2017. Lancet Infectious Diseases17(8): 843-853
Concordance of in vitro and in vivo measures of non-replicating rotavirus vaccine potency
Rotavirus infections remain a leading cause of morbidity and mortality among infants residing in low- and middle-income countries. To address the large need for protection from this vaccine-preventable disease we are developing a trivalent subunit rotavirus vaccine which is currently being evaluated in a multinational Phase 3 clinical trial for prevention of serious rotavirus gastroenteritis. Currently, there are no universally accepted in vivo or in vitro models that allow for correlation of field efficacy to an immune response against serious rotavirus gastroenteritis. As a new generation of non-replicating rotavirus vaccines are developed the lack of an established model for evaluating vaccine efficacy becomes a critical issue related to how vaccine potency and stability can be assessed. Our previous publication described the development of an in vitro ELISA to quantify individual vaccine antigens adsorbed to an aluminum hydroxide adjuvant to address the gap in vaccine potency methods for this non-replicating rotavirus vaccine candidate. In the present study, we report on concordance between ELISA readouts and in vivo immunogenicity in a guinea pig model as it relates to vaccine dosing levels and sensitivity to thermal stress. We found correlation between in vitro ELISA values and neutralizing antibody responses engendered after animal immunization. Furthermore, this in vitro assay could be used to demonstrate the effect of thermal stress on vaccine potency, and such results could be correlated with physicochemical analysis of the recombinant protein antigens. This work demonstrates the suitability of the in vitro ELISA to measure vaccine potency and the correlation of these measurements to an immunologic outcome
Revolution from above in English schools: neoliberalism, the democratic commons and education
The ideas of the New Left and the recently emerged alter-globalisation movements are marginal within current policy debates concerning the English education system. Here I seek to demonstrate the interconnections between the New Left and the alter-globalisation movement and suggest that these ideas contain a powerful corrective to the increasingly authoritarian present. The next part of the article considers the development of neoliberalism both in a theoretical context and since the arrival of the new ConservativeâLiberal government in the UK. Here I outline the rapid transformation of English schools under the academies programme and look at how it has been explicitly linked to ideas of âmoral collapseâ evident in the popular discourse of âBroken Britainâ. Especially significant in this respect has been the labelling of comprehensive schools as âfailuresâ and the explicit imposition of more authoritarian understandings of pedagogy. I seek to explore both the rapidity of this transformation in the context of the dissatisfaction with the idea of comprehensive schools shown by the political Right and the Third Wayâs reworking of socialism. Finally I briefly consider more progressive alternatives for schools and education by returning to the idea of the democratic commons. In this respect, the cultural Left needs to explore more radical alternatives beyond the defence of comprehensive schooling which sounds both nostalgic and misplaced within our global times
Are social innovation paradigms incommensurable?
This paper calls attention to the problematic use of the concept of social innovation which remains undefined despite its proliferation throughout academic and policy discourses. Extant research has thus far failed to capture the socio-political contentions which surround social innovation. This paper therefore draws upon the work of Thomas Kuhn and conducts a paradigmatic analysis of the field of social innovation which identifies two emerging schools: one technocratic, the other democratic. The paper identifies some of the key thinkers in each paradigm and explains how the struggle between these two paradigms reveals itself to be part of a broader conflict between neoliberalism and it opponents and concludes by arguing that future research focused upon local contextualised struggles will reveal which paradigm is in the ascendancy
Rethinking Radical Democracy
Over the course of three decades, vocabularies of radical democracy have pressed their stamp on democratic thought. Trading on the intuition that there is more to democracy than elections, they have generated critical insights into the important role that practices of pluralisation and critique play in bettering institutional politics. As a result, few would today deny the radical democratic contribution to democratic thought. What many might question, however, is its continuing traction. The paper probes this question, focusing on the nuanced place of democracy in contemporary radical work. It grapples with the difficulties that this poses for radical democrats and it suggests that a way of overcoming these difficulties â that threaten to undermine the coherence of radical democracy â is to rethink and reconstruct the distinctiveness of its vocabulary. The paper attempts such a reconstruction. It develops the idea of the promissory rule of the many and it discusses the ways it rejuvenates broader democratic thought
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