21 research outputs found
Inhibition of BMP2 and BMP4 Represses Barrettâs Esophagus While Enhancing the Regeneration of Squamous Epithelium in Preclinical Models
European Research Council (ERC) starting grant: ERC-StG
282079 TargetS4BarretERC-POC 632258 BMP4EACDutch
government grant: LSH-TKI-PPP 2017Plan Estatal de InvestigaciĂłn
CientĂfica y TĂ©cnica y de InnovaciĂłn 2013â2016, ISCIII-SubdirecciĂłn General
de EvaluaciĂłn y Fomento de la InvestigaciĂłnMinisterio de EconomĂa y
Competitividad, Spain (grant PI16/01642 & PI10/01096
Menstrual bloodâderived stromal cells modulate functional properties of mouse and human macrophages
Menstrual blood-derived stromal cells (MenSCs) are emerging as a strong candidate for cell-based
therapies due to their immunomodulatory properties. However, their direct impact on innate immune
populations remains elusive. Since macrophages play a key role in the onset and development of
inflammation, understanding MenSCs implication in the functional properties of these cells is required
to refine their clinical effects during the treatment of inflammatory disorders. In this study, we
assessed the effects that MenSCs had on the recruitment of macrophages and other innate immune
cells in two mouse models of acute inflammation, a thioglycollate (TGC)-elicited peritonitis model
and a monobacterial sepsis model. We found that, in the TGC model, MenSCs injection reduced the
percentage of macrophages recruited to the peritoneum and promoted the generation of peritoneal
immune cell aggregates. In the sepsis model, MenSCs exacerbated infection by diminishing the
recruitment of macrophages and neutrophils to the site of infection and inducing defective bacterial
clearance. Additional in vitro studies confirmed that co-culture with MenSCs impaired macrophage
bactericidal properties, affecting bacterial killing and the production of reactive oxygen intermediates.
Our findings suggest that MenSCs modulate the macrophage population and that this modulation
must be taken into consideration when it comes to future clinical applications.Plan Estatal de InvestigaciĂłn CientĂfica y TĂ©cnica y de InnovaciĂłn 2013-2016, ISCIII-SubdirecciĂłn General de EvaluaciĂłn y Fomento de la InvestigaciĂłn, Ministerio de EconomĂa y Competitividad, Spain
PI16/01642
PI10/01096European Union (EU)Catedra de InvestigaciĂłn Antonio Chamorro-Alejandro Otero, Universidad de Granada
CACH2017-
SLAMF8 Downregulates Mouse Macrophage Microbicidal Mechanisms via PI3K Pathways
SUPPLEMENTARY MATERIAL
The Supplementary Material for this article can be found online at:
https://www.frontiersin.org/articles/10.3389/fimmu.2022.910112/
full#supplementary-materialACKNOWLEDGMENTS
SR-P is a PhD student belonging to the Official Doctoral
Program in Biomedicine of the University of Granada. The
authors thank Dr. Ana Santos Carro and Dr. David Porcell
from the Center of Technical Instrumentation, University of Granada, for their excellent technical assistance with confocal
microscopy, and Dr. M.C. Ruiz-Ruiz and Dr. Silvia Calpe-Flores
for revising the manuscript.Signaling lymphocytic activation molecule family 8 (SLAMF8) is involved in the negative modulation of NADPH oxidase activation. However, the impact of SLAMF8 downregulation on macrophage functionality and the microbicide mechanism remains elusive. To study this in depth, we first analyzed NADPH oxidase activation pathways in wild-type and SLAMF8-deficient macrophages upon different stimulus. Herein, we describe increased phosphorylation of the Erk1/2 and p38 MAP kinases, as well as increased phosphorylation of NADPH oxidase subunits in SLAMF8-deficient macrophages. Furthermore, using specific inhibitors, we observed that specific PI3K inhibition decreased the differences observed between wild-type and SLAMF8-deficient macrophages, stimulated with either PMA, LPS, or Salmonella typhimurium infection. Consequently, SLAMF8-deficient macrophages also showed increased recruitment of small GTPases such as Rab5 and Rab7, and the p47phox subunit to cytoplasmic Salmonella, suggesting an impairment of Salmonella-containing vacuole (SCV) progression in SLAMF8-deficient macrophages. Enhanced iNOS activation, NO production, and IL-6 expression were also observed in the absence of SLAMF8 upon Salmonella infection, either in vivo or in vitro, while overexpression of SLAMF8 in RAW264.7 macrophages showed the opposite phenotype. In addition, SLAMF8-deficient macrophages showed increased activation of Src kinases and reduced SHP-1 phosphate levels upon IFNÎł and Salmonella stimuli in comparison to wild-type macrophages. In agreement with in vitro results, Salmonella clearance was augmented in SLAMF8-deficient mice compared to that in wild-type mice. Therefore, in conclusion, SLAMF8 intervention upon bacterial infection downregulates mouse macrophage activation, and confirmed that SLAMF8 receptor could be a potential therapeutic target for the treatment of severe or unresolved inflammatory conditions.Plan Estatal de InvestigaciĂłn CientĂfica y TĂ©nica y de InnovaciĂłn, ISCIII SubdireccioÌn
General de EvaluacioÌn y Fomento de la InvestigaciĂłn, Ministerio de EconomĂa y Competitividad, Spain
(Grants PI16/01642 and PI10/01096
Decreased breadth of the antibody response to the spike protein of SARS-CoV-2 after repeated vaccination
Introduction: The rapid development of vaccines to prevent COVID-19 has raised the need to compare the capacity of different vaccines in terms of developing a protective humoral response. Previous studies have shown inconsistent results in this area, highlighting the importance of further research to evaluate the efficacy of different vaccines.
Methods: This study utilized a highly sensitive and reliable flow cytometry method to measure the titers of IgG1 isotype antibodies in the blood of healthy volunteers after receiving one or two doses of various vaccines administered in Spain. The method was also used to simultaneously measure the reactivity of antibodies to the S protein of the original Wuhan strain and variants B.1.1.7 (Alpha), B.1.617.2 (Delta), and B.1.617.1 (Kappa).
Results: Significant differences were observed in the titer of anti-S antibodies produced after a first dose of the vaccines ChAdOx1 nCov-19/AstraZeneca, mRNA-1273/Moderna, BNT162b2/Pfizer-BioNTech, and Ad26.COV.S/Janssen. Furthermore, a relative reduction in the reactivity of the sera with the Alpha, Delta, and Kappa variants, compared to the Wuhan strain, was observed after the second boosting immunization.
Discussion: The findings of this study provide a comparison of different vaccines in terms of anti-S antibody generation and cast doubts on the convenience of repeated immunization with the same S protein sequence. The multiplexed capacity of the flow cytometry method utilized in this study allowed for a comprehensive evaluation of the efficacy of various vaccines in generating a protective humoral response. Future research could focus on the implications of these findings for the development of effective COVID-19 vaccination strategies.This work was funded by intramural grant CSIC-COVID19-
004: 202020E081 (to BA) and CSIC-COVID19-004: 202020E165
(to MF). LH has been supported by an FPI fellowship from the
Spanish Ministry of Science and Innovation. IB has been supported
by an H2020-MSCA-ITN-2016 training network grant of the
European Union (GA 721358)
Decreased breadth after boost
The rapid development of vaccines to prevent infection by SARS-CoV-2 virus causing COVID-19 makes necessary to compare the capacity of the different vaccines in terms of development of a protective humoral response. Here, we have used a highly sensitive and reliable flow cytometry method to measure the titers of antibodies of the IgG1 isotype in blood of volunteers after receiving one or two doses of the vaccines being administered in Spain. We took advantage of the multiplexed capacity of the method to measure simultaneously the reactivity of antibodies with the S protein of the original strain Wuhan-1 and the variant B.1.1.7 (Alpha). We found significant differences in the titer of anti-S antibodies produced after a first dose of the vaccines ChAdOx1 nCov-19/AstraZeneca, mRNA-1273/Moderna, BNT162b2/Pfizer-BioNTech and Ad26.COV.S/Janssen. Most important, we found a relative reduction in the reactivity of the sera with the B.1.1.7 versus the Wuhan-1 variant after the second boosting immunization. These data allow to make a comparison of different vaccines in terms of anti-S antibody generation and cast doubts about the convenience of repeatedly immunizing with the same S protein sequence.This work was funded by intramural grant CSIC-COVID19-004: 202020E081 (to B.A.)
and CSIC-COVID19-004: 202020E165 (to MF). L.H has been supported by an FPI fellowship from
the Spanish Ministry of Science and Innovation. I.B. has been supported by an H2020-MSCA-ITN2016 training network grant of the European Union (GA 721358)Peer reviewe
Decreased breadth of the antibody response to the spike protein of SARS-CoV-2 after repeated vaccination
IntroductionThe rapid development of vaccines to prevent COVID-19 has raised the need to compare the capacity of different vaccines in terms of developing a protective humoral response. Previous studies have shown inconsistent results in this area, highlighting the importance of further research to evaluate the efficacy of different vaccines.MethodsThis study utilized a highly sensitive and reliable flow cytometry method to measure the titers of IgG1 isotype antibodies in the blood of healthy volunteers after receiving one or two doses of various vaccines administered in Spain. The method was also used to simultaneously measure the reactivity of antibodies to the S protein of the original Wuhan strain and variants B.1.1.7 (Alpha), B.1.617.2 (Delta), and B.1.617.1 (Kappa).ResultsSignificant differences were observed in the titer of anti-S antibodies produced after a first dose of the vaccines ChAdOx1 nCov-19/AstraZeneca, mRNA-1273/Moderna, BNT162b2/Pfizer-BioNTech, and Ad26.COV.S/Janssen. Furthermore, a relative reduction in the reactivity of the sera with the Alpha, Delta, and Kappa variants, compared to the Wuhan strain, was observed after the second boosting immunization.DiscussionThe findings of this study provide a comparison of different vaccines in terms of anti-S antibody generation and cast doubts on the convenience of repeated immunization with the same S protein sequence. The multiplexed capacity of the flow cytometry method utilized in this study allowed for a comprehensive evaluation of the efficacy of various vaccines in generating a protective humoral response. Future research could focus on the implications of these findings for the development of effective COVID-19 vaccination strategies
Inhibitor of apoptosis proteins, NAIP, cIAP1 and cIAP2 expression during macrophage differentiation and M1/M2 polarization
Monocytes and macrophages constitute the first line of defense of the immune system
against external pathogens. Macrophages have a highly plastic phenotype depending on
environmental conditions; the extremes of this phenotypic spectrum are a pro-inflammatory
defensive role (M1 phenotype) and an anti-inflammatory tissue-repair one (M2 phenotype).
The Inhibitor of Apoptosis (IAP) proteins have important roles in the regulation of several cellular
processes, including innate and adaptive immunity. In this study we have analyzed the
differential expression of the IAPs, NAIP, cIAP1 and cIAP2, during macrophage differentiation
and polarization into M1 or M2. In polarized THP-1 cells and primary human macrophages,
NAIP is abundantly expressed in M2 macrophages, while cIAP1 and cIAP2 show
an inverse pattern of expression in polarized macrophages, with elevated expression levels
of cIAP1 in M2 and cIAP2 preferentially expressed in M1. Interestingly, treatment with the
IAP antagonist SMC-LCL161, induced the upregulation of NAIP in M2, the downregulation
of cIAP1 in M1 and M2 and an induction of cIAP2 in M1 macrophages.This work was supported by Universidad
de Granada, Plan Propio 2015;#P3B: FAM, VMC
(http://investigacion.ugr.es/pages/planpropio/2015/
resoluciones/p3b_def_28072015); Universidad
de Granada CEI BioTic;#CAEP2-84: VMC (http://
biotic.ugr.es/pages/resolucionprovisional
enseaanzapractica22demayo/!); and Canadian nstitutes of Health Research;#231421, #318176,
#361847: STB, ECL, RK (http://www.cihr-irsc.gc.
ca/e/193.html). The funders had no role in study
design, data collection and analysis, decision to
publish, or preparation of the manuscript
Outcomes from elective colorectal cancer surgery during the SARS-CoV-2 pandemic
This study aimed to describe the change in surgical practice and the impact of SARS-CoV-2 on mortality after surgical resection of colorectal cancer during the initial phases of the SARS-CoV-2 pandemic
Human IgG and IgA responses to SARS-CoV-2 vaccination
The emergence of COVID-19 has led to a worldwide challenge for the rapid
development of vaccines. Several types of safe and effective vaccines have been
available in a time frame never seen before. Comparative studies to know the extent of
protection and the immune response elicited by the different vaccines are of outstanding
utility. Here, as a correlate for protection, we perform a comparative study of the
humoral response to three vaccines, ChAdOx1 (Oxford-AstraZeneca), mRNA-1273
(Moderna), and BNT162b2 (Pfizer-BioNTech) by applying a flow cytometry-based
highly sensitive method that we had previously developed. We have found that mRNA
vaccines (mRNA-1273 and BNT162b2) induce a stronger humoral response that lasts
for at least 6 months after vaccination. We also show that only one dose of BNT162b2
is enough to achieve the maximum response in seropositive pre-vaccination donors.This work was funded by intramural grant CSIC-COVID19-004: 202020E081 (to B.A.).N
Vaccine Type-, Age- and Past Infection-Dependence of the Humoral Response to SARS-CoV-2 Spike S Protein
The emergence of COVID-19 has led to a worldwide challenge for the rapid development of vaccines. Several types of safe and effective vaccines have been available in a time frame never seen before. Now that several hundred million people have been vaccinated there is an opportunity to compare vaccines in terms of protection and immune response. Here, we have applied a highly sensitive multiplexed flow cytometry method to measure simultaneously IgM, IgG1 and IgA anti-spike protein antibodies generated in response to three vaccines: ChAdOx1 (Oxford-AstraZeneca), mRNA-1273 (Moderna), and BNT162b2 (Pfizer-BioNTech). We have found that mRNA vaccines (mRNA-1273 and BNT162b2) induce a stronger humoral response, both after the first and the second dose, than the adenovirus-based ChAdOx1 vaccine. We also found that, in the elderly, antibody titers negatively correlate with the age of the donor but, also, that antibody titers remain stable for at least 6 months after complete vaccination. Finally, we found that one dose of BNT162b2 is sufficient to induce the highest antibody titers in seropositive pre-vaccination donors. We hope these data will help to guide future decisions on vaccination strategies.This work was funded by intramural grant CSIC-COVID19-004: 202020E081 (to BA). L.H has been supported by an FPI fellowship from the Spanish Ministry of Science and Innovation.Peer reviewe