bioRxiv Science⌕ Search

Biology subjects

Ghanem, E. N. B.

Publications and source records attributed to Ghanem, E. N. B..

3 recordsLinked to original sources

Host aging induces a senescent-like phenotype in neutrophils and altered transcriptional responses to Streptococcus pneumoniae

Aging drives increased susceptibility to respiratory infections by Streptococcus pneumoniae (pneumococci). Polymorphonuclear leukocytes (PMNs) are among the first responders in the lung following pneumococcal infection and are required for bacterial clearance. However, PMN antimicrobial function declines with age. To identify mechanisms underlying this decline, we performed RNA sequencing on PMNs in the lungs of young and old mice following pulmonary infection with S. pneumoniae. We observed significant transcriptomic differences across host age. Transcriptional analysis followed by functional validation revealed that in infected mice, PMNs from aged hosts failed to upregulate several effector activities including glycolysis and subsequent mitochondrial reactive oxygen species (ROS) production, which are necessary for bacterial killing by PMNs. Analysis of potential transcription factors controlling these changes indicated differential regulation by E2f2 in aged mice, which was linked to lower PMN differentiation resulting in more immature PMNs in the lungs of aged mice compared to young controls. Conversely, PMNs in aged mice displayed a higher senescence-associated secretory phenotype (SASP) score and upregulated pathways involved in cellular senescence. Follow-up functional characterization found that in uninfected hosts, PMNs in aged mice expressed higher levels of SASP factors IL-10, TNF, and ROS, had lower incidence of apoptosis, and had a higher proportion of cells positive for senescence-associated {beta}-galactosidase, features of a senescent-like phenotype. In conclusion, host aging is associated with altered PMN phenotypes, including a shift toward senescent-like energy-deficient cells, which may contribute to impaired host defense and represent potential targets for improved interventions against infection in older adults.

immunology↗

Basal activation of ERK1/2 blunts the antimicrobial activity of neutrophils from aged hosts against antibody opsonized Streptococcus pneumoniae

The decline in neutrophil antimicrobial function renders vaccinated aged hosts less protected against Streptococcus pneumoniae infection. In vaccinated hosts, activation of neutrophils via phagocytic receptors, like complement and Fc{gamma} receptors, mediates bacterial uptake and killing. However, the mechanisms behind changes in signaling downstream of these receptors with age is not known. Mitogen-activated protein kinases (MAPK) are signaling cascades activated in response to cell surface receptors. Using bone marrow neutrophils isolated from young and aged mice, and MAPK phosphorylation array, we found differences in MAPK activation in an opsonin dependent manner (complement vs antibody). Neutrophils from aged mice failed to increase MAPK phosphorylation upon infection with pneumococci opsonized with heat-inactivated (HI) sera from hosts immunized with the pneumococcal conjugate vaccine (PCV), indicating an age-related defect in Fc{gamma}R signaling. Neutrophils from aged mice had higher basal levels of MAPK phosphorylation when compared to young controls, including a 15-fold increase in ERK1/2 phosphorylation. Inhibition of ERK1/2 signaling blunted pneumococcal killing by PMNs from young mice, but improved killing by neutrophils from aged mice, only when the bacteria were opsonized with HI immune but not naive sera. In young human participants, in vitro inhibition of ERK1/2 in neutrophils resulted in decreased pneumococcal killing, but only following PCV vaccination, suggesting that the effects of ERK1/2 inhibition are clinically relevant in vaccinated hosts. This study identifies age-related changes in ERK1/2 activation and demonstrates that balanced activation of this pathway is crucial for neutrophil antimicrobial activity against antibody opsonized bacteria following vaccination in both mice and humans. Summary sentenceImbalanced ERK1/2 activation impairs neutrophil activity in aged hosts. LEGENDS O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=134 SRC="FIGDIR/small/656427v1_ufig1.gif" ALT="Figure 1"> View larger version (21K): org.highwire.dtl.DTLVardef@179c31forg.highwire.dtl.DTLVardef@f95d4borg.highwire.dtl.DTLVardef@15c1dcborg.highwire.dtl.DTLVardef@a8076e_HPS_FORMAT_FIGEXP M_FIG C_FIG Graphical AbstractGraphic Abstract Created in BioRender. Bou Ghanem, E. (2025) https://BioRender.com/jjoodsu.

immunology↗

Neutrophil-driven cardiac damage during invasive Streptococcus pneumoniae infection is regulated by CD73

Streptococcus pneumoniae (pneumococcus)-induced cardiac events are one of the life-threatening infection outcomes of invasive pneumococcal disease. S. pneumoniae has the ability to invade the myocardium and damage cardiomyocytes, however the contribution of the immune response during this process is not fully understood. We previously found that polymorphonuclear cells (PMNs) are crucial for host defense against S. pneumoniae lung infection and that extracellular adenosine (EAD) production, by exonucleosidases CD39 and CD73, controlled the anti-bacterial functions of these cells. The objective of this study was to explore the role of PMNs and the EAD-pathway in host cardiac damage during invasive pneumococcal infection. Upon intra-peritoneal (i.p.) injection with invasive S. pneumoniae TIGR4 strain, hearts of C57BL/6 mice showed an increased influx of PMNs as determined by flow cytometry. However, the increased PMN numbers failed to contain the bacterial burden in the heart and showed positive correlation with serum levels of the cardiac damage marker Troponin-1. Influx of PMNs into the heart was associated with constant presence of neutrophil degranulation products in the cardiac tissue. Depletion of PMNs prior infection reduced pneumococcal burden in the heart and lowered the Troponin-1 levels thus, indicating their role in cardiac damage. While exploring the mechanisms underlying the damaging PMN response, we found that by 24hpi, there was a significant reduction in the expression of CD39 and CD73 on cardiac PMNs. The role of CD73 in regulating cardiac damage was tested in vivo using CD73-/- mice which had significantly higher bacterial burden and cardiac damage compared to wild type mice despite similar PMN numbers. The role of CD73 expression on PMNs was also tested ex vivo using the HL-1 cardiomyocyte cell line which upon S. pneumoniae infection, showed increased cell death in presence of CD73-/- PMNs. Our findings have identified a detrimental role for PMNs in cardiac damage during invasive pneumococcal infection that is in part driven by reduced expression of EAD-producing enzymes in late disease stages.

immunology↗