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Biology subjects

Marinho, C. R. F.

Publications and source records attributed to Marinho, C. R. F..

6 recordsLinked to original sources

Extrafollicular activation of B cells contributes to the long-term survival of CD28KO mice infected with Sylvio X10/4Trypanosoma cruzi parasites

In the initial phase of infections, prior to the development of the follicular antibody response, an extrafollicular B cell response develops in lymphoid organs, with activation of cells displaying B cell receptors with low affinity to the different epitopes of the invading pathogen. To evaluate if this response allows early protection in the murine host, we investigated its role during infection with Sylvio X10/4 (a low virulence Trypanosoma cruzi parasite) of CD28KO mice, a mouse strain that does not develop a follicular antibody response, in parallel with infected C57BL/6 mice (used as control). We observed that T. cruzi infection allowed CD28KO mice to survive for a long period of time, ranging from a few weeks to almost a year. However, the CD28KO mice showed higher levels of subpatent parasitemia than C57BL/6 mice, as well as more intense and long-lasting inflammatory infiltrates in the heart. Following the CD28KO mice throughout the infection, we found a significant reduction in the number of CD8+ T cells in the spleen, but not CD4+ T cells, as well as a strong fluctuation in the number of B cells. Meanwhile, although the production of IFN-{gamma} in the spleen was not affected by CD28 deficiency, a reduction in IL-2 production was observed in anti-CD3 stimulated CD4+ T cells from infected CD28KO mice. Regarding the humoral response, after confirming that the T follicular helper (TFH) cells and germinal center B cells (GC-B) were absent in the infected CD28KO mice, we demonstrated that the extrafollicular antibodies, present in CD28KO infected mice, allowed an early protection in the murine host, since the transfer of serum from chronic CD28KO mice to naive CD28KO mice, which were infected with T. cruzi 24 hours later, promoted a significant reduction in parasitemia. These results demonstrate that low-affinity anti-T cruzi extrafollicular antibodies contribute to the control of circulating parasites, allowing Sylvio X10/4-infected CD28KO mice to survive for a long time. AUTHOR SUMMARYLow-affinity anti-T cruzi extrafollicular antibodies contribute to the control of circulating parasites, allowing Sylvio X10/4-infected CD28KO mice to survive for a long time.

immunology↗

Protein arginylation is regulated during SARS-CoV-2 infection

In 2019, the world witnessed the onset of an unprecedented pandemic. In September 2021, the infection by SARS-CoV-2 had already been responsible for the death of more than 4 million people worldwide. Recently, we and other groups discovered that SARS-CoV-2 infection induces ER-stress and activation of unfolded protein response (UPR) pathway. The degradation of misfolded/unfolded proteins is an essential element of proteostasis and occurs mainly in lysosomes or proteasomes. The N-terminal arginylation of proteins is characterized as an inducer of ubiquitination and proteasomal degradation by the N-end rule pathway. Here we present, for the first time, data on the role of arginylation during SARS-CoV-2 infection. We studied the modulation of protein arginylation in Vero CCL-81 and Calu-3 cells infected after 2h, 6h, 12h, 24h, and 48h. A reanalysis of in vivo and in vitro public omics data combined with immunoblotting was performed to measure the levels of ATE1 and arginylated proteins. This regulation is seen specifically during infections by coronaviruses. We demonstrate that during SARS-CoV-2 infection there is an increase in the expression of the ATE1 enzyme associated with regulated levels of specific arginylated proteins. On the other hand, infected macrophages showed no ATE1 regulation. An important finding revealed that modulation of the N-end rule pathway differs between different types of infected cells. We also confirmed the potential of tannic acid to reduce viral load, and furthermore, to modulate ATE1 levels during infection. In addition, the arginylation inhibitor merbromin (MER) is also capable of both reducing viral load and reducing ATE1 levels. Taken together, these data show the importance of arginylation during the progression of SARS-CoV-2 infection and open the door for future studies that may unravel the role of ATE1 and its inhibitors in pathogen infection.

cell biology↗

SARS-CoV-2 activates ER stress and Unfolded protein response

Coronavirus disease-2019 (COVID-19) pandemic caused by the SARS-CoV-2 coronavirus infection is a major global public health concern affecting millions of people worldwide. The scientific community has joint efforts to provide effective and rapid solutions to this disease. Knowing the molecular, transmission and clinical features of this disease is of paramount importance to develop effective therapeutic and diagnostic tools. Here, we provide evidence that SARS-CoV-2 hijacks the glycosylation biosynthetic, ER-stress and UPR machineries for viral replication using a time-resolved (0-48 hours post infection, hpi) total, membrane as well as glycoproteome mapping and orthogonal validation. We found that SARS-CoV-2 induces ER stress and UPR is observed in Vero and Calu-3 cell lines with activation of the PERK-eIF2-ATF4-CHOP signaling pathway. ER-associated protein upregulation was detected in lung biopsies of COVID-19 patients and associated with survival. At later time points, cell death mechanisms are triggered. The data show that ER stress and UPR pathways are required for SARS-CoV-2 infection, therefore representing a potential target to develop/implement anti-CoVID-19 drugs.

biochemistry↗

Critical contribution of endothelial protein C receptor in experimental malaria-associated acute respiratory distress syndrome

The severity of Plasmodium falciparum malaria is associated with parasite cytoadherence, but there is limited knowledge about the effect of parasite cytoadherence in malaria-associated acute respiratory distress syndrome (ARDS). Our objective was to evaluate the cytoadherence of infected red blood cells (iRBCs) in a murine model of ARDS and to appraise a potential function of endothelial protein C receptor (EPCR) in ARDS pathogenesis. DBA/2 mice infected with P. berghei ANKA were classified as ARDS- or hyperparasitemia (HP)-developing mice according to respiratory parameters and parasitemia. Lungs, blood and bronchoalveolar lavage were collected for gene expression or protein analyses. Primary cultures of microvascular lung endothelial cells from DBA/2 mice were analyzed for iRBC interactions. Lungs from ARDS-developing mice showed evidence of iRBC accumulation along with an increase in EPCR and TNF concentrations. Furthermore, TNF increased iRBC adherence in vitro. Dexamethasone-treated infected mice showed low levels of TNF and EPCR mRNA expression and, finally, decreased vascular permeability, thus protecting mice from ARDS. In conclusion, we identified that increased iRBC cytoadherence in the lungs underlies malaria-associated ARDS in DBA/2-infected mice and that inflammation increased cytoadherence capacity, suggesting a participation of EPCR and a conceivable target for drug development.

pathology↗

Malaria during pregnancy and newborns outcome in an unstable transmission area in Brazil: a population-based record linkage study

BackgroundMalaria during pregnancy is one of the major causes of mortality in tropical regions, causing maternal anemia, intrauterine growth retardation, preterm birth, and low birth weight (LBW). The integration of the information systems is crucial to assess the dimension of gestational malaria in a wide and useful way, to improve decision making and maternal-child health.\n\nMethods and FindingsAn observational population-based study acquired information retrospectively from all live births that occurred between 2006 and 2014 in Cruzeiro do Sul (Acre, Brazil). Social and clinical data of the mother and newborn was extracted from the Information System of Live Births. Malaria episodes information was obtained from the Brazilian Epidemiological Surveillance Information System Malaria. A deterministic record linkage was performed to assess malaria impact on pregnancy. The studied population presented a malaria incidence of 8.9%, of which 63.9% infected by Plasmodium (P.) vivax. Reduction of newborns birth weight at term (small for gestational age (SGA) and LBW) has been found associated with P. vivax infection during pregnancy (SGA - OR 1.24, 95% CI 1.02-1.52, p=0.035; term LBW - OR 1.39, 95% CI 1.03-1.88, p=0.033). Additionally, P. falciparum infection during pregnancy has been found to be associated with preterm births (OR 1.54, 95% CI 1.09-2.18, p=0.016), which is related with late preterm births (OR 1.59, 95% CI 1.11-2.27, p=0.011).\n\nConclusionsDespite the decrease of malaria cases during the evaluated period, we present evidence of the deleterious effects of gestational malaria in a low transmission area in the Amazonian region. Regardless of Plasmodium species, malaria during pregnancy poses a risk for newborns birth weight reduction, highlighting the impact that P. vivax has on the fetus.\n\nFundingSao Paulo Research Foundation - FAPESP/Brazil.

epidemiology↗

Plasmodium falciparum infection during pregnancy impairs fetal head growth: prospective and populational-based retrospective studies

BackgroundMalaria in pregnancy is associated with adverse effects on the fetus and newborns. However, the outcome on a newborns head circumference (HC) is still unclear. Here, we show the relation of malaria during pregnancy with fetal head growth.\n\nMethodsClinical and anthropometric data were collected from babies in two cohort studies of malaria-infected and non-infected pregnant women, in the Brazilian Amazon. One enrolled prospectively (PCS, Jan. 2013 to April 2015) through volunteer sampling, and followed until delivery, 600 malaria-infected and non-infected pregnant women. The other assembled retrospectively (RCS, Jan. 2012 to Dec. 2013) clinical and malaria data from 4697 pregnant women selected through population-based sampling. The effects of malaria during pregnancy in the newborns were assessed using a multivariate logistic regression. According with World Health Organization guidelines babies were classified in small head (HC < 1 SD below the median) and microcephaly (HC < 2 SD below the median) using international HC standards.\n\nResultsAnalysis of 251 (PCS) and 232 (RCS) malaria-infected, and 158 (PCS) and 3650 (RCS) non-infected women with clinical data and anthropometric measures of their babies was performed. Among the newborns, 70 (17.1%) in the PCS and 934 (24.1%) in the RCS presented with a small head (SH). Of these, 15 (3.7%) and 161 (4.2%), respectively, showed microcephaly (MC). The prevalence of newborns with a SH (30.7% in PCS and 36.6% in RCS) and MC (8.1% in PCS and 7.3% in RCS) was higher among babies born from women infected with Plasmodium falciparum during pregnancy. Multivariate logistic regression analyses revealed that P. falciparum infection during pregnancy represents a significant increased odds for the occurrence of a SH in newborns (PCS: OR 3.15, 95% CI 1.52-6.53, p=0.002; RCS: OR 1.91, 95% CI 1.21-3.04, p=0.006). Similarly, there is an increased odds of MC in babies born from mothers that were P. falciparum-infected (PCS: OR 5.09, 95% CI 1.12-23.17, p=0.035). Moreover, characterization of placental pathology corroborates the association analysis, particularly through the occurrence of more syncytial nuclear aggregates and inflammatory infiltrates in placentas from babies with the reduced head circumference.\n\nConclusionsThis work indicates that falciparum-malaria during pregnancy presents an increased likelihood of occurring reduction of head circumference in newborns, which is associated with placental malaria.\n\nTrial Registrationregistered as RBR-3yrqfq in the Brazilian Clinical Trials Registry

epidemiology↗