Suppression by allogeneic-specific regulatory T cells is dependent on the degree of HLA compatibility

Copyright © 2021 The Authors. This article is distributed under the terms of the CC BY 4.0 Unported license.Regulatory T cell (Treg) infusion for graft-versus-host disease treatment has been increasingly investigated. However, polyclonal Treg may suppress the desired graft-versus-leukemia effect. Although allogeneic-specific (allo-specific) Treg may provide a more-targeted graft-versus-host disease treatment, there is the need to develop easily translatable expansion protocols and to better characterize their specificity and mechanisms of suppression. In this article, we provide a robust protocol for human allo-specific Treg expansion and characterize their phenotype, potency, and specificity of suppression by testing different expansion conditions and suppression assay milieus. We found that higher concentrations of IL-2 during expansion with allogeneic APC yielded allo-specific Treg that were more-potent suppressors and displayed a more activated phenotype. Although responses to the same APC present during expansion were the most suppressed, responses to third-party APC partially matched to the expansion APC were still significantly more suppressed than responses to fully mismatched APC. Furthermore, suppression of responses to the expansion APC was strictly contact dependent, whereas suppression of responses to mismatched APC was partially independent of contact. Finally, distinct subsets in fresh and expanded Treg could be described using multidimensional visualization techniques. We propose that allo-specific Treg are HLA specific and that the mechanisms of suppression elicited depend on their compatibility with the stimulators.This work was supported by Fundação para a Ciência e Tecnologia, Portugal, under the Harvard Medical School–Portugal Program Project Induction of Immune Tolerance in Human Allogeneic Hematopoietic Stem Cell Transplantation (HMSP-ICT/0001/2011) and Gilead Sciences, under Programa Gilead GÉNESE (FPJ001262). J.B. received a Ph.D. grant from Fundação para a Ciência e Tecnologia (PD/BD/105769/2014).info:eu-repo/semantics/publishedVersio

Mesenchymal stromal cells induce regulatory T cells via epigenetic conversion of human conventional CD4 T cells in vitro

© 2020 The Authors. S TEM CELLS published by Wiley Periodicals LLC on behalf of AlphaMed Press. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.Regulatory T cells (Treg) play a critical role in immune tolerance. The scarcity of Treg therapy clinical trials in humans has been largely due to the difficulty in obtaining sufficient Treg numbers. We performed a preclinical investigation on the potential of mesenchymal stromal cells (MSCs) to expand Treg in vitro to support future clinical trials. Human peripheral blood mononuclear cells from healthy donors were cocultured with allogeneic bone marrow-derived MSCs expanded under xenogeneic-free conditions. Our data show an increase in the counts and frequency of CD4+ CD25high Foxp3+ CD127low Treg cells (4- and 6-fold, respectively) after a 14-day coculture. However, natural Treg do not proliferate in coculture with MSCs. When purified conventional CD4 T cells (Tcon) are cocultured with MSCs, only cells that acquire a Treg-like phenotype proliferate. These MSC-induced Treg-like cells also resemble Treg functionally, since they suppress autologous Tcon proliferation. Importantly, the DNA methylation profile of MSC-induced Treg-like cells more closely resembles that of natural Treg than of Tcon, indicating that this population is stable. The expression of PD-1 is higher in Treg-like cells than in Tcon, whereas the frequency of PDL-1 increases in MSCs after coculture. TGF-β levels are also significantly increased MSC cocultures. Overall, our data suggest that Treg enrichment by MSCs results from Tcon conversion into Treg-like cells, rather than to expansion of natural Treg, possibly through mechanisms involving TGF-β and/or PD-1/PDL-1 expression. This MSC-induced Treg population closely resembles natural Treg in terms of phenotype, suppressive ability, and methylation profile.This project received funding from: Fundação para a Ciência e Tecnologia, Portugal, under the Harvard Medical School-Portugal Program project Induction of Immune Tolerance in Human Allogeneic Hematopoietic Stem Cell Transplantation (HMSP-ICT/0001/2011) and UID/BIM/50005/2019, project funded by Fundação para a Ciência e a Tecnologia (FCT)/Ministério da Ciência, Tecnologia e Ensino Superior (MCTES) through Fundos do Orçamento de Estado. We also acknowledge the funding received from POR Lisboa 2020 through the project PRECISE – Accelerating progress toward the new era of precision medicine (project no. 16394).info:eu-repo/semantics/publishedVersio

Chemical study and biological activity evaluation of two Azorean Macroalgae: Ulva rigida and Gelidium microdon

Copyright: © 2013 Silva M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credite.The green macroalga Ulva rigida C. Agardh (Chlorophyta) and the red macroalga Gelidium microdon Kützing (Rhodophyta), collected from the Azorean archipelago, were investigated for their secondary metabolites and their in vitro growth inhibitory effect on three human tumor cell lines: MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer) and A375-C5 (melanoma), as well as for their antifungal and antibacterial activities. The methanol extract of U. rigida furnished isofucosterol (1), 7(E)-3β-hydroxy-5α,6α-epoxymegastigmane (2) and (+)-dehydrovomifoliol (3) while the methanol extract of G. microdon yielded cholesterol (4) and lumichrome (5). The crude extracts of both macroalgae were found to be moderately active against the three cell lines whereas compound 1 showed a weak effect and compound 2 was inactive. The crude extracts of the two macroalgae were found to be moderately active against some fungi and bacteria while compounds 1 and 2 were inactive against all microorganisms tested

Steroid–quinoline hybrids for disruption and reversion of protein aggregation processes

Reversing protein aggregation within cells may be an important tool to fight protein-misfolding disorders such as Alzheimer’s, Parkinson’s, and cardiovascular diseases. Here we report the design and synthesis of a family of steroid−quinoline hybrid compounds based on the framework combination approach. This set of hybrid compounds effectively inhibited Aβ1−42 self-aggregation in vitro by delaying the exponential growth phase and/or reducing the quantity of fibrils in the steady state. Their disaggregation efficacy was further demonstrated against preaggregated Aβ1−42 peptides in cellular assays upon their endocytosis by neuroblastoma cells, as they reverted both the number and the average area of fibrils back to basal levels. The antiaggregation effect of these hybrids was further tested and demonstrated in a cellular model of general protein aggregation expressing a protein aggregation fluorescent sensor. Together, our results show that the new cholesterol−quinoline hybrids possess wide and marked disaggregation capacities and are therefore promising templates for the development of new drugs to deal with conformational disorders.Thanks are due to the University of Aveiro, FCT/MEC, Centro 2020 and Portugal2020, the COMPETE Program, and the European Union (FEDER Program) via the financial support to the research units LAQV-REQUIMTE (UIDB/50006/2020), IBiMED (UID/BIM/04501/2019) and CICECO- Aveiro Institute of Materials (UID/CTM/50011/2019), financed by national funds through the FCT/MCTES, to the Portuguese NMR Network, to the ThiMES Project (POCI-01- 0145-FEDER-016630), and to the PAGE Project “Protein Aggregation Across the Lifespan” (CENTRO-01-0145- FEDER-000003), including postdoctoral grants to H.M.T.A. (BPD/UI98/4861/2017) and R.N.d.S. (BPD/UI98/6327/2018). M.P. was supported by Ph.D. Grant SFRH/BD/135655/2018. A.R.S. and S.G. were supported by national funds (OE) through FCT, I.P., in the scope of the framework contract foreseen in numbers 4, 5, and 6 of Article 23 of the Decree-Law 57/2016 of August 29, changed by Law 57/2017 of July 19. Microphotographs were acquired in the LiM facility of iBiMED/UA, a member of the Portuguese Platform of BioImaging (PPBI) (POCI-01-0145-FEDER-022122).info:eu-repo/semantics/publishedVersio

Accuracy of the new rapid test for monitoring adalimumab levels

The loss of response to adalimumab (ADL) has been related to low serum concentrations at trough. Currently, most methods commercially available for the quantification of ADL are enzyme-linked immunosorbent assay (ELISA) based, with a turnaround time of approximately 8 h, delaying the target dosage adjustment to the subsequent infusion. In this study, we aimed to evaluate the performance of the newly available rapid-test ADL quantification assay by comparing it with three established ELISA methods, using spiked samples and a set of clinical samples.info:eu-repo/semantics/publishedVersio

Descrição da biodiversidade terrestre e marinha dos Açores

1. Os Açores são um arquipélago isolado de nove ilhas oceânicas, pertence à região biogeográfica da Macaronésia e está entre as regiões mais ricas em fungos, plantas e animais da Europa. Este capítulo destaca o que sabemos sobre os fungos, a fauna e a flora dos habitats terrestres, dulçaquícolas e marinhos dos Açores. 2. Neste capítulo, são apresentadas as estimativas do número total de espécies e subespécies conhecidas actualmente nos Açores. Todos os grupos taxonómicos terrestres mais importantes foram analisados: fungos, líquenes, diatomáceas dulçaquícolas, briófitos (musgos, hepáticas e antocerotas), plantas vasculares (licófitas, fetos, gimnospérmicas e angiospérmicas), platelmintes (vermes), nemátodos, anelídeos (minhocas), moluscos terrestres (lesmas e caracóis), artrópodes (insectos, aracnídeos, milípedes, etc.) e vertebrados (peixes de água doce, anfíbios, répteis, aves e mamíferos). A presente obra inclui também espécies do ambiente marinho, como as algas (macroalgas), a maioria dos filos de invertebrados do litoral e os vertebrados marinhos (répteis, peixes e mamíferos). As listas de espécies e subespécies (Capítulos 2-15) são baseadas nos taxa identificados numa grande variedade de publicações, tendo essa informação sido compilada por um vasto grupo de especialistas. 3. Actualmente, o número total de taxa (espécies e subespécies) terrestres nos Açores está estimado em cerca de 6164 (cerca de 6112 espécies). A inclusão de uma listagem exaustiva das espécies de aves não-nidificantes e de uma listagem preliminar de espécies de aves potencialmente nidificantes acrescenta 325 espécies e subespécies ao total das espécies açorianas. 4. O número total de espécies e subespécies endémicas terrestres dos Açores é de cerca de 452 (411 espécies). Os animais são os mais diversos em endemismos, com 331 taxa (Arthropoda = 266; Mollusca = 49; Vertebrata = 14; Nematoda = 2), compreendendo cerca de 73% dos endemismos terrestres dos Açores. A percentagem de endemismo nos Mollusca (44%) é notável. As plantas vasculares contam com 73 endemismos, os Fungi (incluindo os líquenes) têm 34 e, tanto as diatomáceas dulçaquícolas como os briófitos, incluem sete espécies endémicas. 5. Quando comparada com os arquipélagos vizinhos da Macaronésia (Madeira e Canárias), a fauna e flora terrestres dos Açores é caracterizada por uma menor taxa de endemismo, de apenas 7%, contrastando com os cerca de 20% para a Madeira e de 30% para as Canárias. 6. No que diz respeito aos organismos marinhos, são listados 1883 taxa pertencentes a 16 filos. O número total de espécies e subespécies marinhas endémicas dos Açores é de cerca de 39, a maior parte delas moluscos (29 espécies). 7. O número total de taxa terrestres e marinhos (espécies e subespécies) nos Açores, está estimado em cerca de 8047. Os organismos marinhos agora listados, perfazem cerca de 23% da biodiversidade dos Açores. 8. O número total de taxa terrestres e marinhos (espécies e subespécies) endémicos dos Açores está estimado em cerca de 491.ABSTRACT: 1. The Azores is a remote oceanic archipelago of nine islands which belongs to the Macaronesia biogeographical region and is among the richest regions concerning fungi, plant and animal diversity in Europe. This chapter highlights what we know about the Azorean terrestrial, freshwater and marine Fungi, Flora and Fauna. 2. In this chapter we summarize the current estimates of the total number of species and subspecies presently known to occur in the Azores. The most important terrestrial taxonomic groups were studied: Fungi, Lichens, Bacillariophyta (freshwater diatoms), Bryophyta sensu lato (mosses, liverworts and hornworts), vascular plants – Tracheobionta, including Lycopodiophyta (quillworts), Pteridophyta (ferns), Pinophyta (gymnosperms) and Magnoliophyta (angiosperms), Platyhelminthes (flatworms), Nematoda (roundworms), Annelida (earthworms), Terrestrial Mollusca (slugs and snails), Arthropoda (millipedes, centipedes, mites, spiders, insects, etc.) and Vertebrata (freshwater fishes, amphibians, reptiles, birds and mammals). In addition, we expand this list to the marine realm, including Algae (macroalgae), coastal invertebrates (most Phyla) and marine vertebrates (fishes, reptiles and mammals). The list of species and subspecies (Chapters 2 to 15) is based on the taxa recognized in primary published literature sources, compiled by a vast group of experts. 3. Currently the total number of terrestrial taxa (species and subspecies) in the Azores is estimated of about 6164 (about 6112 species). The inclusion of an exhaustive listing of non breeding species and a preliminary list of potentially breeding species adds 325 species and subspecies of birds to the Azorean list of species. 4. The total number of terrestrial endemic species and/or subspecies from the Azores is about 452 (411 species). Animals are the most represented in this respect, with 331 taxa (Arthropoda = 266; Mollusca = 49; Vertebrata = 14; Nematoda = 2), that is, about 73% of the Azorean terrestrial endemics. The percentage of endemism within Mollusca (44%) is remarkable. Vascular plants have 73 endemic taxa, while Fungi (including Lichens) have 34, freshwater diatoms and bryophytes have seven endemic species each. 5. Compared to the other nearest Macaronesian archipelagos (Madeira and Canaries), the Azorean terrestrial fauna and flora is characterized by a lower percentage of endemism (only 7%, which contrasts with nearly 20% for Madeira and 30% for the Canary islands). 6. Concerning the marine organisms, we listed about 1883 taxa belonging to 16 Phyla. The total number of marine endemic species and/or subspecies from the Azores is about 39, most of them being molluscs (29 species). 7. Currently, the total number of terrestrial and marine taxa (species and subspecies) in the Azores is estimated in about 8047. The marine organisms currently listed make up about 23% of the Azorean biodiversity. 8. Currently, the total number of terrestrial and marine endemic taxa (species and subspecies) in the Azores is estimated of about 491

Electron and Positron Scattering Cross Sections from CO\u3csub\u3e2\u3c/sub\u3e: A Comparative Study over a Broad Energy Range (0.1−5000 eV)

In this Review, we present a comparative study between electron and positron scattering cross sections from CO2 molecules over a broad impact energy range (0.1−5000 eV). For electron scattering, new total electron scattering cross sections (e-TCS) have been measured with a high resolution magnetically confined electron beam transmission system from 1 to 200 eV. Dissociative electron attachment processes for electron energies from 3 to 52 eV have been analyzed by measuring the relative O−anion production yield. In addition, elastic, inelastic, and total scattering cross section calculations have been carried out in the framework of the Independent Atom Model by using the Screening Corrected Additive Rule, including interference effects (IAM-SCARI). Based on the previous cross section compilation from Itikawa (J. Phys. Chem. Ref. Data, 2002, 31, 749−767) and the present measurements and calculations, an updated recommended e-TCS data set has been used as reference values to obtain a self-consistent integral cross section data set for the elastic and inelastic (vibrational excitation, electronic excitation, and ionization) scattering channels. A similar calculation has been carried out for positrons, which shows important differences between the electron scattering behavior: e.g., more relevance of the target polarization at the lower energies, more efficient excitation of the target at intermediate energies, but a lower total scattering cross section for increasing energies, even at 5000 eV. This result does not agree with the charge independence of the scattering cross section predicted by the first Born approximation (FBA). However, we have shown that the inelastic channels follow the FBA’s predictions for energies above 500 eV while the elastic part, due to the different signs of the scattering potential constituent terms, remains lower for positrons even at the maximum impact energy considered here (5000 eV). As in the case of electrons, a self-consistent set of integral positron scattering cross sections, including elastic and inelastic (vibrational excitation, electronic excitation, positronium formation, and ionization) channels is provided. Again, to derive these data, positron scattering total cross sections based on a previous compilation from Brunger et al. (J. Phys. Chem. Ref. Data, 2017, 46, 023102) and the present calculation have been used as reference values. Data for the main inelastic channels, i.e. direct ionization and positronium formation, derived with this procedure, show excellent agreement with the experimental results available in the literature. Inconsistencies found between different model potential calculations, both for the elastic and inelastic collision processes, suggest that new calculations using more sophisticated methods are required

A Comparative Study over a Broad Energy Range (0.1-5000 eV)

Funding Information: This study has been partially supported by the Spanish Ministerio de Ciencia e Innovación (Project PID2019-104727RB-C21) and the Spanish Ministerio de Universidades (Project PRX21/00340). Work performed at Lawrence Berkeley National Laboratory was supported by the US Department of Energy, Office of Science, Division of Chemical Sciences of the Office of Basic Energy Sciences, under Contract DE-AC02-05CH11231. M.H. was supported by the US Department of Energy Office of Science, Basic Energy Sciences under Award No. DE-SC0019482. P.L.V. acknowledges the Portuguese National Funding Agency (FCT) through Research Grants CEFITEC (UIDB/00068/2020) and PTDC/FIS-AQM/31281/2017. The work is part of COST Action CA18212 - Molecular Dynamics in the GAS phase (MD-GAS). Publisher Copyright: © 2022 American Chemical Society.In this Review, we present a comparative study between electron and positron scattering cross sections from CO2 molecules over a broad impact energy range (0.1-5000 eV). For electron scattering, new total electron scattering cross sections (e-TCS) have been measured with a high resolution magnetically confined electron beam transmission system from 1 to 200 eV. Dissociative electron attachment processes for electron energies from 3 to 52 eV have been analyzed by measuring the relative O- anion production yield. In addition, elastic, inelastic, and total scattering cross section calculations have been carried out in the framework of the Independent Atom Model by using the Screening Corrected Additive Rule, including interference effects (IAM-SCARI). Based on the previous cross section compilation from Itikawa (J. Phys. Chem. Ref. Data, 2002, 31, 749-767) and the present measurements and calculations, an updated recommended e-TCS data set has been used as reference values to obtain a self-consistent integral cross section data set for the elastic and inelastic (vibrational excitation, electronic excitation, and ionization) scattering channels. A similar calculation has been carried out for positrons, which shows important differences between the electron scattering behavior: e.g., more relevance of the target polarization at the lower energies, more efficient excitation of the target at intermediate energies, but a lower total scattering cross section for increasing energies, even at 5000 eV. This result does not agree with the charge independence of the scattering cross section predicted by the first Born approximation (FBA). However, we have shown that the inelastic channels follow the FBA's predictions for energies above 500 eV while the elastic part, due to the different signs of the scattering potential constituent terms, remains lower for positrons even at the maximum impact energy considered here (5000 eV). As in the case of electrons, a self-consistent set of integral positron scattering cross sections, including elastic and inelastic (vibrational excitation, electronic excitation, positronium formation, and ionization) channels is provided. Again, to derive these data, positron scattering total cross sections based on a previous compilation from Brunger et al. (J. Phys. Chem. Ref. Data, 2017, 46, 023102) and the present calculation have been used as reference values. Data for the main inelastic channels, i.e. direct ionization and positronium formation, derived with this procedure, show excellent agreement with the experimental results available in the literature. Inconsistencies found between different model potential calculations, both for the elastic and inelastic collision processes, suggest that new calculations using more sophisticated methods are required.publishersversionpublishe

Isolation and functional characterization of proinflammatory acidic phospholipase A2 from Bothrops leucurus snake venom

AbstractIn the present study, an acidic PLA2, designated Bl-PLA2, was isolated from Bothrops leucurus snake venom through two chromatographic steps: ion-exchange on CM-Sepharose and hydrophobic chromatography on Phenyl-Sepharose. Bl-PLA2 was homogeneous on SDS-PAGE and when submitted to 2D electrophoresis the molecular mass was 15,000Da and pI was 5.4. Its N-terminal sequence revealed a high homology with other Asp49 acidic PLA2s from snake venoms. Its specific activity was 159.9U/mg and the indirect hemolytic activity was also higher than that of the crude venom. Bl-PLA2 induced low myotoxic and edema activities as compared to those of the crude venom. Moreover, the enzyme was able to induce increments in IL-12p40, TNF-α, IL-1β and IL-6 levels and no variation of IL-8 and IL-10 in human PBMC stimulated in vitro, suggesting that Bl-PLA2 induces proinflammatory cytokine production by human mononuclear cells. Bothrops leucurus venom is still not extensively explored and knowledge of its components will contribute for a better understanding of its action mechanism