Spn Infection Research Articles

Abstract We062: Characterization of Transcriptional Responses to Streptococcus pneumoniae Infection in the Heart versus Sterile Cardiac Injury

Background: Streptococcus pneumoniae ( Spn ) remains the leading cause of community-acquired pneumonia (CAP). As many as 30% of patients hospitalized for CAP experience a major adverse cardiac event (MACE). Importantly, the elevated risk for MACE is not only limited to the time of acute infection but also extends several years post-hospitalization. Research Question: Is the host response to Spn- mediated injury in the heart different from the response to sterile injury? Goals: To identify defining features of the host response to Spn in the heart via differences in gene expression. Methods: We performed a bioinformatic analysis of bulk RNAseq data collected from hearts of mice acutely infected with Spn and compared this to publicly available data sets of mice that received experimental myocardial infarction, i.e. ischemia reperfusion and permanent ligation. Using DeSeq2, we identified 2,971 genes with differential expression between the infected and infarcted conditions. To identify gene expression differences that might be relevant to human patients, we subsequently used the Reactome GSA software to map our differential expression results onto predicted human pathways. Results: Categorization of the gene roles identified those in the classical antibody-mediated complement activation as the dominant pathway with differential expression. Reactome analysis of the top 5 differentially expressed genes (p < 0.05) at the individual level identified BAIAP2L1 , CXCL9 , IRF7 , IL18BP , RPL12 , and PSMB10 as significantly upregulated in Spn infected hearts versus infarcted hearts. As one feature of Spn mediated damage is the recruitment of immune cells to the affected sites and cardiac remodeling, we validated upregulation of CXCL9 as well as CXCL10, potent recruiters of T-cells, as enhanced during Spn infection. Conclusions: Our analyses provide information on how the host response to disseminated pneumococcal infection in the heart is distinct from sterile injury and identified mediators that may be contributing to adverse cardiac remodeling.

A novel Bacillus aerolatus CX253 attenuates inflammation induced by Streptococcus pneumoniae in childhood and pregnant rats by regulating gut microbiome

Streptococcus pneumoniae (Spn) is the predominant pathogen responsible for community-acquired pneumonia (CAP) in children under five years old, and it can induce over 17% of pregnant women. However, no more effective measures exist to prevent infection induced by Spn in these two special populations. The beneficial microbes can antagonize Spn and provide new targets for preventing pneumococcal infections. This study used 16S rRNA gene sequencing and targeted metabolomics to evaluate the role of the Bacillus aerolatus CX253 (CX253) in alleviating Spn infection. Additionally, the colonization of CX253 was observed in nose, trachea, and lung by using confocal laser scanning microscopy and fluorescent labeling techniques. Compared with the model group, the expression level of interleukin-1β was dropped 1.81-fold and 2.22-fold, and interleukin-6 was decreased 2.39-fold and 1.84-fold. The express of tumor necrosis factor-α was down 2.30-fold and 3.84-fold in prevention group of childhood and pregnant rats, respectively. The 16S rRNA sequencing results showed that CX253 administration alone significantly increased the abundance of Lactobacillus, Limosilactobacillus, and Prevotella in the gut of childhood and pregnant rats. Furthermore, the CX253 increased propionate in the gut of childhood rats and increased propionate and butyrate in the gut of pregnant rats to inhibit pulmonary inflammation. In summary, CX253 attenuated Spn-induced inflammation by regulating the gut microbiota and SCFAs. The research provides valuable information for the prevention of pneumonia.Graphical

A bacterial vesicle-based pneumococcal vaccine against influenza-mediated secondary Streptococcus pneumoniae pulmonary infection

Streptococcus pneumoniae (Spn) is a common pathogen causing a secondary bacterial infection following influenza which leads to severe morbidity and mortality during seasonal and pandemic influenza. Therefore, there is an urgent need to develop bacterial vaccines that prevent severe post-influenza bacterial pneumonia. Here, an improved Yersinia pseudotuberculosis strain (designated as YptbS46) possessing an Asd+ plasmid pSMV92 could synthesize high amounts of the Spn pneumococcal surface protein A (PspA) antigen and monophosphoryl lipid A as an adjuvant. The recombinant strain produced outer membrane vesicles (OMVs) enclosing a high amount of PspA protein (designated as OMV-PspA). A prime-boost intramuscular immunization with OMV-PspA induced both memory adaptive and innate immune responses in vaccinated mice, reduced the viral and bacterial burden, and provided complete protection against influenza-mediated secondary Spn infection. Also, the OMV-PspA immunization afforded significant cross-protection against the secondary Spn A66.1 infection and long-term protection against the secondary Spn D39 challenge. Our study implies that an OMV vaccine delivering Spn antigens can be a new promising pneumococcal vaccine candidate.

NAD+ metabolism is a key modulator of bacterial respiratory epithelial infections

Lower respiratory tract infections caused by Streptococcus pneumoniae (Spn) are a leading cause of death globally. Here we investigate the bronchial epithelial cellular response to Spn infection on a transcriptomic, proteomic and metabolic level. We found the NAD+ salvage pathway to be dysregulated upon infection in a cell line model, primary human lung tissue and in vivo in rodents, leading to a reduced production of NAD+. Knockdown of NAD+ salvage enzymes (NAMPT, NMNAT1) increased bacterial replication. NAD+ treatment of Spn inhibited its growth while growth of other respiratory pathogens improved. Boosting NAD+ production increased NAD+ levels in immortalized and primary cells and decreased bacterial replication upon infection. NAD+ treatment of Spn dysregulated the bacterial metabolism and reduced intrabacterial ATP. Enhancing the bacterial ATP metabolism abolished the antibacterial effect of NAD+. Thus, we identified the NAD+ salvage pathway as an antibacterial pathway in Spn infections, predicting an antibacterial mechanism of NAD+.

Streptococcus pneumoniaecoinfection results in increased disease in a murine model of influenza B virus infection

Abstract Secondary Streptococcus pneumoniae (Spn) infection is commonly associated with increased hospitalization and death following seasonal influenza virus infection. The animal models of influenza A virus (IAV) and secondary Spn coinfection have described the increased morbidity and mortality leading to several proposed mechanisms of enhanced disease. Influenza B virus (IBV) causes 25% of seasonal influenza cases and is associated with bacterial coinfection, yet little is known about the immune response to IBV and Spn coinfection. Here, we established a mouse model of IBV and Spn coinfection to study disease in singly and coinfected animals and define mechanism(s) of enhanced disease. Like IAV and Spn coinfection, infection with IBV followed by Spn inoculation resulted in increased morbidity and mortality compared to IBV or Spn infection alone. In IBV primed mice, Spn titers were increased early following bacterial inoculation and Spn rapidly migrated to the lung. Analysis of cytokines in the lung showed increased pro-inflammatory cytokine (TNF-α, IL-1β, and IL-6) responses with secondary Spn infection, which may be associated with enhanced disease. These data suggest that IBV infection initiates innate immune responses that are rapidly enhanced with Spn infection, leading to worsening disease. Ongoing studies include analysis of infiltrating cells during coinfection and histopathologic analysis to complement the innate response data to define mediators of clinical disease. NIH CIDER Contract #75N93021C00018

Age-dependent differences in efferocytosis determine the outcome of opsonophagocytic protection from invasive pathogens

In early life, susceptibility to invasive infection skews toward a small subset of microbes, whereas other pathogens associated with diseases later in life, including Streptococcus pneumoniae (Spn), are uncommon among neonates. To delineate mechanisms behind age-dependent susceptibility, we compared age-specific mouse models of invasive Spn infection. We show enhanced CD11b-dependent opsonophagocytosis by neonatal neutrophils improved protection against Spn during early life. The augmented function of neonatal neutrophils was mediated by higher CD11b surface expression at the population level due to dampened efferocytosis, which also resulted in more CD11bhi "aged" neutrophils in peripheral blood. Dampened efferocytosis during early life could be attributed to the lack of CD169+ macrophages in neonates and reduced systemic expressions of multiple efferocytic mediators, including MerTK. On experimentally impairing efferocytosis later in life, CD11bhi neutrophils increased and protection against Spn improved. Our findings reveal how age-dependent differences in efferocytosis determine infection outcome through the modulation of CD11b-driven opsonophagocytosis and immunity.

Cephalosporins-induced intestinal dysbiosis exacerbated pulmonary endothelial barrier disruption in streptococcus pneumoniae-infected mice.

Antibiotic abuse is growing more severe in clinic, and even short-term antibiotic treatment can cause long-term gut dysbiosis, which may promote the development and aggravation of diseases. Cephalosporins as the broad-spectrum antibiotics are widely used for prevention and treatment of community-acquired respiratory tract infection in children. However, their potential consequences in health and disease have not been fully elaborated. In this study, the effects of cefaclor, cefdinir and cefixime on intestinal microbiota and lung injury were investigated in Streptococcus pneumoniae (Spn)-infected mice. The results showed that the proportion of coccus and bacillus in intestinal microbiota were changed after oral administration with cefaclor, cefdinir and cefixime twice for 10 days, respectively. Compared with the Spn-infected group, the proportion of Bifidobacterium and Lactobacillus in intestine were significantly reduced, while Enterococcus and Candida was increased after cephalosporin treatment. Furthermore, 3 cephalosporins could obviously increase the number of total cells, neutrophils and lymphocytes in BALF as well as the serum levels of endotoxin, IL-2, IL-1β, IL-6 and TNF-α. Mechanically, cephalosporins accelerated Spn-induced pulmonary barrier dysfunction via mediating the mRNA expressions of endothelial barrier-related proteins (Claudin 5, Occludin, and ZO-1) and inflammation-related proteins (TLR4, p38 and NF-κB). However, all of those consequences could be partly reversed by Bifidobacterium bifidum treatment, which was closely related to the elevated acetate production, indicating the protective effects of probiotic against antibiotic-induced intestinal dysbiosis. Therefore, the present study demonstrated that oral administration with cephalosporins not only disrupted intestinal microecological homeostasis, but also increased the risk of Spn infection, resulting in severer respiratory inflammation and higher bacterial loads in mice.

Neutrophil Recruitment in Pneumococcal Pneumonia.

Streptococcus pneumoniae (Spn) is the primary agent of community-acquired pneumonia. Neutrophils are innate immune cells that are essential for bacterial clearance during pneumococcal pneumonia but can also do harm to host tissue. Neutrophil migration in pneumococcal pneumonia is therefore a major determinant of host disease outcomes. During Spn infection, detection of the bacterium leads to an increase in proinflammatory signals and subsequent expression of integrins and ligands on both the neutrophil as well as endothelial and epithelial cells. These integrins and ligands mediate the tethering and migration of the neutrophil from the bloodstream to the site of infection. A gradient of host-derived and bacterial-derived chemoattractants contribute to targeted movement of neutrophils. During pneumococcal pneumonia, neutrophils are rapidly recruited to the pulmonary space, but studies show that some of the canonical neutrophil migratory machinery is dispensable. Investigation of neutrophil migration is necessary for us to understand the dynamics of pneumococcal infection. Here, we summarize what is known about the pathways that lead to migration of the neutrophil from the capillaries to the lung during pneumococcal infection.

Secondary infection with Streptococcus pneumoniae decreases influenza virus replication and is linked to severe disease.

ABSTRACTSecondary bacterial infection is a common complication in severe influenza virus infections. During the H1N1 pandemic of 2009, increased mortality was observed among healthy young adults due to secondary bacterial pneumonia, one of the most frequent bacterial species being Streptococcus pneumoniae (Spn). Previous studies in mice and ferrets have suggested a synergistic relationship between Spn and influenza viruses. In this study, the ferret model was used to examine whether secondary Spn infection (strains BHN97 and D39) influence replication and airborne transmission of the 2009 pandemic H1N1 virus (H1N1pdm09). Secondary infection with Spn after H1N1pdm09 infection consistently resulted in a significant decrease in viral titers in the ferret nasal washes. While secondary Spn infection appeared to negatively impact influenza virus replication, animals precolonized with Spn were equally susceptible to H1N1pdm09 airborne transmission. In line with previous work, ferrets with preceding H1N1pdm09 and secondary Spn infection had increased bacterial loads and more severe clinical symptoms as compared to animals infected with H1N1pdm09 or Spn alone. Interestingly, the donor animals that displayed the most severe clinical symptoms had reduced airborne transmission of H1N1pdm09. Based on these data, we propose an asymmetrical relationship between these two pathogens, rather than a synergistic one, since secondary bacterial infection enhances Spn colonization and pathogenesis but decreases viral titers.

Microbicidal Activity of Hypothiocyanite against Pneumococcus.

Infections caused by Streptococcus pneumoniae (pneumococcus, Spn) manifest in several forms such as pneumonia, meningitis, sinusitis or otitis media and are associated with severe morbidity and mortality worldwide. While current vaccines and antibiotics are available to treat Spn infections, the rise of antibiotic resistance and limitations of the vaccines to only certain Spn serotypes urge the development of novel treatments against Spn. Hypothiocyanite (OSCN-) is a natural antimicrobial product produced by the body’s own innate immune system to fight a variety of pathogens. We recently showed that OSCN- is also capable of killing Spn in vitro. OSCN- is an oxidative agent attacking microbes in a nonspecific manner, is safe for the host and also has anti-inflammatory effects that make it an ideal candidate to treat a variety of infections in humans. However, OSCN- has a short life span that makes its use, dosage and administration more problematic. This minireview discusses the antimicrobial mechanism of action of OSCN- against Spn and elaborates on the potential therapeutic use of OSCN- against Spn and other infectious agents, either alone or in combination with other therapeutic approaches.

Exploration of Recombinant Fusion Proteins YAPO and YAPL as Carrier Proteins for Glycoconjugate Vaccine Design against Streptococcus pneumoniae Infection.

Pneumolysin (Ply), pneumococcal surface protein A (PspA), and pneumococcal surface adhesin A (PsaA) are promising cell surface protein antigen targets for Streptococcus pneumoniae (Spn) vaccine development. Herein, we designed and recombined two fusion proteins, named YAPO and YAPL, which contained the main antigenic epitopes of Ply, PspA, and PsaA. In-depth immunological evaluations revealed that YAPO and YAPL had strong immunocompetence to be well-qualified potential carrier proteins. To verify this possibility, a serotype 3 Spn (ST3) CPS pentasaccharide was conjugated to each fusion protein to generate the resultant glycoconjugates. Immunological studies in mice revealed that, as compared with TT conjugate, YAPO and YAPL conjugates provoked robust T-cell dependent immune responses that could provide better recognition, in vitro efficient opsonophagocytosis, and in vivo effective protection against various serotypes of Spn. Collectively, YAPO and YAPL were identified as immunopotentiating carriers that could help convert immunologically inactive ST3 pentasaccharide into a T cell-dependent antigen and provide efficient and broad spectrum of immunoprotection coverage so as to formulate functional glycoconjugate vaccines against Spn infections.

Isotype-specific outcomes in Fc gamma receptor targeting of PspA using fusion proteins as a vaccination strategy against Streptococcus pneumoniae infection

Streptococcus pneumoniae (Spn) remains a considerable threat to public health despite the availability of antibiotics and polysaccharide conjugate vaccines. The lack of mucosal immunity in addition to capsular polysaccharide diversity, has proved to be problematic in developing a universal vaccine against Spn. Targeting antigen to Fc receptors is an attractive way to augment both innate and adaptive immunity against mucosal pathogens, by promoting interactions with activating Fcγ receptors (FcγR) that mediate diverse immunomodulatory functions. The effect of targeting FcγR is highly influenced by the IgG subclass, which bares differential affinities for activating and inhibitory FcγR. In the current study we demonstrate targeting activating FcγR with fusion proteins consisting of PspA and IgG2a Fc enhance PspA-specific immune responses, and effectively protect against mucosal Spn challenge. Specifically, targeting PspA to FcγR polarized alveolar macrophage to the AM1 phenotype and increased conventional dendritic cell subsets in the lung in addition to augmenting Th1 cytokines and PspA-specific IgG and IgA. In contrast, fusion proteins consisting of PspA fused to the IgG1 Fc provided minimal benefit over administration of PspA alone, as a result of interaction with the inhibitory FcγRIIB. Protective efficacy of the IgG1 fusion protein was significantly enhanced in animals deficient for FcγRIIB accompanied by increased B cell maturation and proliferation levels in these animals. These studies demonstrate FcγR targeting is an effective strategy for inducing potent cellular and humoral responses via mucosal immunization with Fc fusion proteins, however, careful consideration of the Fc region utilized is required since Fc isotype subclass heavily influenced immunization induced effector functions and survival against lethal Spn challenge. Fc-engineering with specific attention to FcγRIIB engagement presents a valuable vaccine strategy for protecting against Spn infection.

Double Deletion of C5aR1 and C5aR2 during Streptococcal Pneumonia

Abstract The complement system is essential for the clearance of encapsulated bacteria such as Streptococcus pneumoniae (Spn), recognized as a major pathogen of pneumonia. Yet, it is not entirely clear whether C5 and its activation product C5a have beneficial or detrimental effects on the outcome of Spn infection. C5a binds to its two homologous C5a receptors (C5aR1 and C5aR2). The extent of functional overlap and role distribution between C5aR1 and C5aR2 remains enigmatic. A critical barrier to allow studies on the role distributions of C5aRs has been the absence of a mouse strain with dual deletion of both receptors - which are encoded as adjacent, linked genes on mouse chromosome 7. CRISPR/Cas9 guided gene editing was combined with zygote/pronucleus microinjections in C57BL/6J mice to generate a C5aR1/C5aR2 double-knockout strain (C5aR1/2−/−). PCR-based screening and Sanger sequencing confirmed a 12.6 kB deletion of the coding regions of C5aR1 and C5aR2 of the new mouse strain, C57BL/6J-Del( 7C5ar2-C5ar1)1Bosm. Phenotyping of C5aR1/2−/− mice was followed by functional studies including intra-tracheal injections of recombinant mouse C5a or intranasal injections of Spn TIGR4. The airway influx of Ly6G+SiglecF- neutrophils by C5a was abrogated in C5aR1/2−/− mice. Surprisingly, the dual genetic absence of C5a receptors was associated with a higher build-up of neutrophil numbers in alveolar spaces after Spn TIGR4 infection. In a transcriptome-wide screen, FACS-sorted myeloid cells from infected lungs of double and single knockout mice were subjected to RNA-sequencing. Identifying novel differentially expressed genes in the C5aR1/2−/− mice during Spn infection will help to uncover potential synergisms and redundancies of C5aR1 and C5aR2.

Isotype-specific outcomes in Fc gamma receptor targeting of PspA using fusion proteins as a vaccination strategy against Streptococcus pneumoniae infection

Abstract Streptococcus pneumoniae (Spn) remains a considerable threat to public health despite the availability of antibiotics and polysaccharide conjugate vaccines. The lack of mucosal immunity in addition to capsular polysaccharide diversity, has proved to be problematic in developing a universal vaccine against Spn. Targeting antigen to Fc receptors is an attractive way to augment both innate and adaptive immunity against mucosal pathogens. The effect of targeting Fcγ receptors (FcγR) is highly influenced by the IgG subclass, which bares differential affinities for activating and inhibitory FcγR. In the current study we demonstrate targeting activating FcγR with fusion proteins consisting of PspA and IgG2a Fc enhance PspA-specific immune responses, and effectively protect against mucosal Spn challenge. Specifically, targeting PspA to FcyR polarized alveolar macrophage to the AM1 phenotype and increased conventional dendritic cell subsets in the lung in addition to augmenting Th1 cytokines and PspA-specific IgG and IgA. In contrast, fusion proteins consisting of PspA fused to the IgG1 Fc provided minimal benefit as a result of interaction with the inhibitory FcγRIIB. Protective efficacy of the IgG1 fusion protein was significantly enhanced in animals deficient for FcγRIIB accompanied by increased B cell maturation and proliferation levels in these animals. These studies demonstrate FcγR targeting is an effective strategy for inducing potent cellular and humoral responses via mucosal immunization with Fc fusion proteins, however, careful consideration of the Fc region utilized, is required. Fc-engineering with specific attention to FcγRIIB engagement presents a valuable vaccine strategy for protecting against Spn infection.

Streptococcus pneumoniae coinfection dictates susceptibility to oral Listeria monocytogenes through suppressive Natural Killer (NK) cell activity

Abstract Coinfection with viral and bacterial pathogens is a major health concern and results in significant morbidity and mortality. Recent research has aimed to understand the complex interplay between multiple pathogens and the immune response during viral:bacterial coinfections. However, studies using bacterial-bacterial coinfections are lacking. Streptococcus pneumoniae (Spn) and Listeria monocytogenes (Lm) are both bacterial pathogens that are capable of causing severe infections in young, elderly, and immunocompromised populations. Here we show that coinfection with intratracheal (IT) Spn increases susceptibility of typically resistant mice to oral Lm infection. On day 3 post infection (p.i.), the presence of Spn increased oral Lm burdens in the spleen, liver, and lungs of coinfected C57/BL6 mice. In addition, while the anti-inflammatory cytokine Interleukin-10 (IL-10) is limited during oral Lm infection, Spn infection induced the production of IL-10 from Natural Killer (NK) cells, and that this correlated with an increase in Lm burdens during coinfection. To assess the importance of NK cell IL-10 in the absence of an ongoing infection, mice were inoculated with a recombinant Lm polypeptide, designated L1S, which stimulates NK cell IL-10 production. In the presence of L1S, the induction of NK cell IL-10 similarly increased systemic burdens of oral Lm. Lastly, BMMs stimulated with rIL-10 and infected with Lm have increased bacterial burdens when compared to untreated but infected controls. Overall, our data show that coinfection impairs host protection through the induction of NK cell production of IL-10 and dictates susceptibility to oral Lm infection.

Excessive Reactive Oxygen Species Inhibit IL-17A+ γδ T Cells and Innate Cellular Responses to Bacterial Lung Infection.

Aims: Excessive reactive oxygen species (ROS) are detrimental to immune cellular functions that control pathogenic microbes; however, the mechanisms are poorly understood. Our aim was to determine the immunological consequences of increased ROS levels during acute bacterial infection. Results: We used a model of Streptococcus pneumoniae (Spn) lung infection and superoxide dismutase 3-deficient (SOD3-/-) mice, as SOD3 is a major antioxidant enzyme that catalyses the dismutation of superoxide radicals. First, we observed that in vitro, macrophages from SOD3-/- mice generated excessive phagosomal ROS during acute bacterial infection. In vivo, there was a significant reduction in infiltrating neutrophils in the bronchoalveolar lavage fluid and reduced peribronchial and alveoli inflammation in SOD3-/- mice 2 days after Spn infection. Annexin V/propidium iodide staining revealed enhanced apoptosis in neutrophils from Spn-infected SOD3-/- mice. In addition, SOD3-/- mice showed an altered macrophage phenotypic profile, with markedly diminished recruitment of monocytes (CD11clo, CD11bhi) in the airways. Further investigation revealed significantly lower levels of the monocyte chemokine CCL-2, and cytokines IL-23, IL-1β, and IL-17A in Spn-infected SOD3-/- mice. There were also significantly fewer IL-17A-expressing gamma-delta T cells (γδ T cells) in the lungs of Spn-infected SOD3-/- mice. Innovation: Our data demonstrate that SOD3 deficiency leads to an accumulation of phagosomal ROS levels that initiate early neutrophil apoptosis during pneumococcal infection. Consequent to these events, there was a failure to initiate innate γδ T cell responses. Conclusion: These studies offer new cellular and mechanistic insights into how excessive ROS can regulate innate immune responses to bacterial infection.

Recent Advances in Pathogenic Streptococcus Vaccine Development.

Streptococcus pneumoniae (Spn) and Streptococcus pyogenes (Spy) cause many invasive and noninvasive diseases responsible for high morbidity and mortality worldwide. Safe, efficacious and affordable vaccines could have a significant, positive impact on the global infectious disease burden. Since the implementation of pneumococcal vaccine in the 1980s, the incidence of Spn infection has decreased significantly. Still so, these currently used multivalent polysaccharides and conjugated pneumococcal vaccines have some limitations. For Spy, there are even no vaccines available yet. There is an urgent need of new vaccines against Spn and Spy. Encouragingly, with the hard work of many investigators worldwide, a number of new vaccines candidates are developed with promising results. Of them, many have already entered the clinical trial stage. This review will describe the current status of Spn and Spy vaccine development, with particular focus on protein-based strategy.

Role of Inflammatory Risk Factors in the Pathogenesis of Streptococcus pneumoniae.

Streptococcus pneumoniae (Spn) is a colonizer of the human nasopharynx (NP), causing a variety of infections in humans including otitis media, pneumonia, sepsis, and meningitis. The NP is an immune permissive site which allows for the persistence of commensal bacteria. Acute or chronic respiratory airway inflammation constitutes a significant risk factor for the manifestation of Spn infections. The inflammatory conditions caused by an upper respiratory viral infection or respiratory conditions such as allergic asthma and chronic obstructive pulmonary disorders (COPDs) are implicated in the dysregulation of airway inflammation and tissue damage, which compromise the respiratory barrier integrity. These immune events promote bacterial outgrowth leading to Spn dissemination and invasion into the bloodstream. Therefore, suppression of inflammation and restoration of respiratory barrier integrity could contain Spn infections manifesting in the backdrop of an inflammatory disease condition. The gained knowledge could be harnessed in the design of novel therapeutic interventions to circumvent Spn bacterial infections.

Immunogenicity and protective efficacy of monovalent PCVs containing 22F and 33F polysaccharides in mouse models of colonization and co-infection

BackgroundIn the current transmission, we studied the immunogenicity and protective efficacy of serotypes 22F and 33F in the prevention of colonization and of invasive Streptococcus pneumoniae (Spn) pathogenesis during an influenza co-infection. Serotypes 22F and 33F are emerging Spn serotypes, which are not part of currently administered pneumococcal conjugate vaccine formulations (PCVs). Spn serotype 6A is an ingredient in the currently administered PCV13 vaccine and was therefore included in the study as a control. MethodsAdult (six weeks) and infant (two weeks) C57BL/6 mice were intranasally infected in the nasopharynx (NP) with Spn serotypes 22F, 33F, or 6A. Influenza A H1N1 A/Puerto Rico/8/193 virus (PR8) was introduced one day after the NP Spn colonization. In an immunization challenge study, mice were vaccinated with monovalent 22F, 33F, or 6A polysaccharide conjugated to the CRM197 antigen. The immunized mice were colonized or co-infected to study the vaccines efficacy. ResultsAll three Spn serotypes established colonization in adult and infant mice. The co-infected mice showed an increase in Spn NP density. Invasive Spn infection (bacteremia) was observed following the co-infection with serotypes 22F and 6A but not 33F in adult mice, whereas infant mice developed bacteremia following co-infection with all three Spn serotypes. The vaccinations led to robust serum antibody responses to capsular polysaccharides 22F, 6A, and less for 33F. The vaccinations resulted in reductions of Spn NP colonization density for all three serotypes, prevention of bacteremia, and increased survival with Spn serotypes 22F and 6A. Passive transfer of antisera was associated with a reduction of Spn colonization densities in infant mice. ConclusionVaccinations with monovalent 22F, 33F, or 6A formulations protect against Spn colonization, and the efficacy of the 22F vaccination was comparable to the 6A vaccination in preventing an invasive Spn bacterial infection during an influenza co-infection.

A double edge function of type 17 immunity in Streptococcus pneumoniae pathogenesis during a co-infection with Influenza A Virus

Abstract Streptococcus pneumoniae (Spn) is a respiratory pathogen responsible for significant global disease burden (otitis media, pneumonia, sepsis). Despite success with current Pneumococcal Conjugate Vaccines, a significant proportion of humans remain colonized with Spn, rendering them at risk for developing Spn infections. Upper respiratory nasopharyngeal (NP) colonization by Spn is a precursor to disease, and viral induced inflammation is a key risk factor for the transition of commensal colonization to disease pathogenesis. We utilized a mouse co-infection model of Spn and influenza virus to determine the role of virus induced acute inflammation in the transition of Spn colonization to disease pathogenesis. We show that early Spn NP colonization is allowed by an immune quiescence state, maintained by the lack of IL-17 expression. Introduction of influenza virus in Spn colonized mice elicits a robust IL-17 driven inflammatory response, which induces robust neutrophilia and tissue damage in the NP. The IL-17 mediated inflammatory response was associated with a significant increase in Spn NP bacterial density, leading to Spn invasion and bacteremia. Antibody mediated neutralization of IL-17 suppressed NP inflammation and bacterial density and prevented bacteremia. IL-17 receptor expression was also detected in nasal myeloid cell infiltrate. RoRγt-KO mice remained protected from bacteremia, mimicking the reduced pathogenesis of IL-17 neutralized WT mice. Using RoRγtGFP mutant mice, we detected the recruitment of GFP expressing RoRγt cell population in NP lavages and nasal associated lymphoid tissues. We conclude that IL-17 blockade could contain influenza induced NP pathology and protect against Spn invasive pathogenesis.