To evaluate the effect of XG-102 (formerly D-JNKI1), a TAT-coupled dextrogyre peptide that selectively inhibits the c-Jun N-terminal kinase, in the treatment of endotoxin-induced uveitis (EIU).
EIU ...was induced in Lewis rats by LPS injection. XG-102 was administered at the time of LPS challenge. The ocular biodistribution of XG-102 was evaluated using immunodetection at 24 hours after either 20 microg/kg IV (IV) or 0.2 microg/injection intravitreous (IVT) administrations in healthy or uveitic eyes. The effect of XG-102 on EIU was evaluated using clinical scoring, infiltration cell quantification, inducible nitric oxide synthase (iNOS) expression and immunohistochemistry, and cytokines and chemokines kinetics at 6, 24, and 48 hours using multiplex analysis on ocular media. Control EIU eyes received vehicle injection IV or IVT. The effect of XG-102 on c-Jun phosphorylation in EIU was evaluated by Western blot in eye tissues.
After IVT injection, XG-102 was internalized in epithelial cells from iris/ciliary body and retina and in glial and microglial cells in both healthy and uveitic eyes. After IV injection, XG-102 was concentrated primarily in inflammatory cells of uveitic eyes. Using both routes of administration, XG-102 significantly inhibited clinical signs of EIU, intraocular cell infiltration, and iNOS expression together with reduced phosphorylation of c-Jun. The anti-inflammatory effect of XG-102 was mediated by iNOS, IFN-gamma, IL-2, and IL-13.
This is the first evidence that interfering with the JNK pathway can reduce intraocular inflammation. Local administration of XG-102, a clinically evaluated peptide, may have potential for treating uveitis.
Single-domain antibodies specific to methotrexate (MTX) were obtained after immunization of one llama (
Llama glama). Specific VHH domains (V–D–J-REGION) were selected by panning from an immune-llama ...library using phage display technology. The antibody fragments specific to MTX were purified from
Escherichia coli (C41 strain) periplasm by immobilized metal affinity chromatography with an expression level of around 10
mg/L. A single band around 16,000
Da corresponding to VHH fragments was found after analysis by SDS-PAGE and Western blotting, while competition ELISA demonstrated selective binding to soluble MTX. Surface plasmon resonance (SPR) analysis showed that anti-MTX VHH domains had affinities in the nanomolar range (29–515
nM) to MTX-serum albumin conjugates. The genes encoding anti-MTX VHH were found by IMGT/V-QUEST to be similar to the previously reported llama and human IGHV germline genes. The V–D and D–J junction rearrangements in the seven anti-MTX CDR3 sequences indicate that they were originated from three distinct progenitor B cells. Our results demonstrate that camelid single-domain antibodies are capable of high affinity binding to low molecular weight hydrosoluble haptens. Furthermore, these anti-MTX VHH give new insights on how the antigen binding repertoire of llama single-domain antibody can provide combining sites to haptens in the absence of a VL. This type of single-domain antibodies offers advantages compared to murine recombinant antibodies in terms of production rate and sequence similarity to the human IGHV3 subgroup genes.
FcγRIII (CD16) plays an important role in the anti-tumor effects of therapeutic antibodies. Bi-specific antibodies (bsAbs) targeting FcγRIII represent a powerful alternative to the recruitment of the ...receptor via the Fc fragment, but are not efficiently produced. Single-domain antibodies (sdAbs) endowed with many valuable structural features might help to bypass this problem. In the present work, we have isolated anti-FcγRIII sdAbs (C21 and C28) from a phage library generated from a llama immunized with FcγRIIIB extra-cellular domains. These sdAbs bind FcγRIIIAsup+ NK cells and FcγRIIIBsup+ polymorphonuclear cells, but not FcγRIsup+ or FcγRIIsup+ cells, as detected by indirect immunofluorescence. Competition experiments showed that C21 and C28 sdAbs bind different FcγRIII epitopes, with C21 recognizing a linear and C28 a conformational epitope of the receptor. Surface plasmon resonance experiments showed that C21 and C28 sdAbs bind FcγRIII with a KsubD in the 10 and 80 nM range, respectively. Importantly, the engagement by both molecules of FcγRIIIA expressed by transfected Jurkat T cells or by NK cells derived from peripheral blood induced a strong IL-2 and IFN-γ production, respectively. These anti-FcγRIII sdAbs represent versatile tools for generating bsAbs under various formats, able to recruit FcγRIII killer cells to target and destroy tumor cells.
FcgammaRIII (CD16) plays an important role in the anti-tumor effects of therapeutic antibodies. Bi-specific antibodies (bsAbs) targeting FcgammaRIII represent a powerful alternative to the ...recruitment of the receptor via the Fc fragment, but are not efficiently produced. Single-domain antibodies (sdAbs) endowed with many valuable structural features might help to bypass this problem. In the present work, we have isolated anti-FcgammaRIII sdAbs (C21 and C28) from a phage library generated from a llama immunized with FcgammaRIIIB extra-cellular domains. These sdAbs bind FcgammaRIIIA+ NK cells and FcgammaRIIIB+ polymorphonuclear cells, but not FcgammaRI+ or FcgammaRII+ cells, as detected by indirect immunofluorescence. Competition experiments showed that C21 and C28 sdAbs bind different FcgammaRIII epitopes, with C21 recognizing a linear and C28 a conformational epitope of the receptor. Surface plasmon resonance experiments showed that C21 and C28 sdAbs bind FcgammaRIII with a K(D) in the 10 and 80 nM range, respectively. Importantly, the engagement by both molecules of FcgammaRIIIA expressed by transfected Jurkat T cells or by NK cells derived from peripheral blood induced a strong IL-2 and IFN-gamma production, respectively. These anti-FcgammaRIII sdAbs represent versatile tools for generating bsAbs under various formats, able to recruit FcgammaRIII killer cells to target and destroy tumor cells.
Presst-Next, the European study carried out between 2004 and 2006, highlighted the importance of teamwork in reducing the frequency of professional exhaustion and the early departure of care staff. ...On the basis of these results, the Armand-Trousseau Hospital in Paris (AP-HP 75) has committed to a participative ergonomics approach at its paediatric emergency service.
We present the results of the XMM-Newton and NuSTAR observations taken as part of the ongoing, intensive multi-wavelength monitoring program of the Seyfert 1 galaxy Mrk 817 by the AGN Space Telescope ...and Optical Reverberation Mapping 2 (AGN STORM 2) Project. The campaign revealed an unexpected and transient obscuring outflow, never before seen in this source. Of our four XMM-Newton/NuSTAR epochs, one fortuitously taken during a bright X-ray state has strong narrow absorption lines in the high-resolution grating spectra. From these absorption features, we determine that the obscurer is in fact a multi-phase ionized wind with an outflow velocity of \(\sim\)5200 km s\(^{-1}\), and for the first time find evidence for a lower ionization component with the same velocity observed in absorption features in the contemporaneous HST spectra. This indicates that the UV absorption troughs may be due to dense clumps embedded in diffuse, higher ionization gas responsible for the X-ray absorption lines of the same velocity. We observe variability in the shape of the absorption lines on timescales of hours, placing the variable component at roughly 1000 \(R_g\) if attributed to transverse motion along the line of sight. This estimate aligns with independent UV measurements of the distance to the obscurer suggesting an accretion disk wind at the inner broad line region. We estimate that it takes roughly 200 days for the outflow to travel from the disk to our line of sight, consistent with the timescale of the outflow's column density variations throughout the campaign.
Les historiens ont longtemps privilégié le facteur technique dans l’approche des révolutions industrielles. Dans cette logique monocausale, le progrès technique était assimilé à une succession ...d’inventions apparues dans des secteurs pionniers, moteurs de la croissance, entraînant le reste de l’économie, dite traditionnelle, dans son sillage. L’un des paradoxes de cette approche consistait à valoriser l’innovation tout en évitant d’interroger les pratiques inventives. La dynamique interne du progrès technique et les traits de génie des inventeurs tenaient lieu de modèles explicatifs. La remise en cause de ces approches suscite bien des interrogations de méthode. Comment repérer les formes de l’invention ordinaire, en cerner les acteurs ? Comment assigner une origine à des nouveautés dont l’antériorité se perd dans la mémoire commune ? Comment appréhender des savoirs pratiques instables et non codifiés que ne livrent pas les corpus constitués de sources ? Comment concilier les définitions construites de l’invention et de l’inventeur, les catégories déjà forgées par les institutions et le corps social, et les mentions informelles ou indirectes de l’invention ? Ces questions débordent l’écrit. Cet ouvrage, issu d’un colloque international tenu à Paris en 2003, élargit le concept de sources : au-delà des « sources-textes », il considère les dessins, les enregistrements sonores, les instruments et outils, les installations, les échantillons, les modèles, les prototypes, etc. Il propose une réflexion originale sur le statut des archives de l’invention, sur leur mode de production et sur les méthodologies mises en œuvre dans leur exploitation.
FcgRIII (CD16) plays an important role in the anti-tumor effects of therapeutic antibodies. Bi-specific anti-bodies (bsAbs) targeting FcgRIII represent a powerful alternative to the recruitment of ...the receptor via the Fc fragment, but are not efficiently produced. Single-domain antibodies (sdAbs) endowed with many valuable structural features might help to bypass this problem. In the present work, we have isolated anti-FcgRIII sdAbs (C21 and C28) from a phage library generated from a llama immunized with FcgRIIIB extra-cellular domains. These sdAbs bind FcgRIIIA 1 NK cells and FcgRIIIB 1 poly-morphonuclear cells, but not FcgRI 1 or FcgRII 1 cells, as detected by indirect immunofluorescence. Competition experiments showed that C21 and C28 sdAbs bind different FcgRIII epitopes, with C21 recognizing a linear and C28 a conformational epitope of the receptor. Surface plasmon resonance experiments showed that C21 and C28 sdAbs bind FcgRIII with a K D in the 10 and 80 nM range, respectively. Importantly, the engagement by both molecules of FcgRIIIA expressed by transfected Jurkat T cells or by NK cells derived from peripheral blood induced a strong IL-2 and IFN-g production, respectively. These anti-FcgRIII sdAbs represent versatile tools for generating bsAbs under various formats, able to recruit FcgRIII killer cells to target and destroy tumor cells.
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