Regulation of miR-61 and col-19 via TGF-ß and Notch signalling in Caenorhabditis elegans against Klebsiella aerogenes infection.

Klebsiella aerogenes, previously known as Enterobacter aerogenes, is a gram-negative bacterium typically present in the gastrointestinal tract. While numerous studies reported the pathogenicity and drug resistance of this bacterium there remains a lack of comprehensive research on K. aerogenes induced alterations in the host cellular mechanisms. In this study, we identify a previously uncharacterized C. elegans miR-61 that defines an evolutionarily conserved miRNA important for development and innate immunity regulation through Notch and TGF-ß signaling pathway. We employed C. elegans wild-type (N2) as well as mutant strains, such as TGF-ß (sma-6) and notch-signaling pathway mutants (adm-4 and mir-61). Our results have demonstrated that the K. aerogenes infected mutants exhibited significantly reduced survival rate, reduced pharyngeal pumping, altered swimming and chemotactic behavior. Moreover, K. aerogenes affects the healthspan by increasing ROS level in the mutants. The gene expression analysis revealed that K. aerogenes upregulated egl-30, tph-1 and sod-1 in adm-4, mir-61 mutants not in sma-6. The in-silico analysis indicated an interaction between mir-61 and col-19, which was confirmed by the upregulation of miR-61 expression and the downregulation of col-19 in sma-6, adm-4, and wild-type strains. These findings suggest that C. elegans activates mir-61 and col-19 regulation through the Notch and TGF-ß signaling pathway against K. aerogenes infection.

Cronobacter sakazakii Cue for the Attraction and Its Impact on the Immunity of Caenorhabditis elegans.

Cronobacter sakazakii, an opportunistic foodborne pathogen prevalently detected in contaminated powdered infant formula, is associated with different diseases, including meningitis. It can cross the blood-brain barrier and affects the CNS. The impact of C. sakazakii on host neuronal cells and behavior is largely unknown. Hence, detailed molecular data are required to understand its severity. Caenorhabditis elegans is a unique model for studying chemical communication, as it relies on chemosensation for searching nutritional supplements. Although, C. sakazakii is pathogenic to C. elegans, our analysis indicated that C. elegans was highly attracted toward C. sakazakii compared to its food source, E. coli OP50. To study the cue for the attraction, bioactive components (RNA/Protein/Lipopolysaccharides/Metabolites) of C. sakazakii were isolated and used for observing the chemotaxis behavior of C. elegans. The results signified that C. elegans was more attracted toward acid extracted metabolites than those of the other extraction methods. The combined action of acid extracted metabolites of C. sakazakii and a candidate pathogen drastically reduced the survival of C. elegans. In addition, qPCR analysis suggested that the exposure of isolated metabolites through acid extraction to C. elegans for 24 h modified the candidate immune regulatory genes involved in pathogen recognition and kinase activity such as clec-60, clec-87, lys-7, akt-2, pkc-1, and jnk-1.

Reduced Reelin Expression Induces Memory Deficits through Dab-1/ NMDAR Signaling Pathway: Cronobacter sakazakii Infection in a Rat Model of Experimental Meningitis.

The purpose of this study was to examine whether the Cronobacter sakazakii infection-induced inflammation alters the Reelin signaling pathway that is involved in learning and memory. To test this, postnatal day (PND)-15 rat pups were either treated with Luria Bertani broth/Escherichia coli OP50/C. sakazakii through oral gavage or maintained as control and allowed to stay with their mothers until PND-24. Experimental groups' rats were subjected to long-term novel object recognition test during their adolescent age PND-30-32. Observed behavioral data showed that C. sakazakii infection causes a deficit in recognition of novel objects from known objects. Further, our analysis showed that C. sakazakii infection-mediated inflammation decreases the Reelin expression by proteolytic cleavage and alters its receptor apolipoprotein E-receptor (ApoER)-2 splice variants ApoER2 (ex19) and ApoER2 (Δ). Subsequently, downregulated Reelin alters the phosphorylation of disabled adapter protein (Dab)-1 and leads to differential expression of N-methyl-D-aspartate (NMDA) receptor subunits 2A and 2B. Further, the NMDA receptor influences the expression of postsynaptic density (PSD)-95 protein and brain-derived neurotrophic factor (BDNF). Observed results suggest a deficit in recognition of novel objects possibly due to the alternation in Reelin signaling pathway.

Proteomic analysis of Caenorhabditis elegans against Salmonella Typhi toxic proteins.

Bacterial effector molecules are crucial infectious agents that can cause pathogenesis. In the present study, the pathogenesis of toxic Salmonella enterica serovar Typhi (S. Typhi) proteins on the model host Caenorhabditis elegans was investigated by exploring the host's regulatory proteins during infection through the quantitative proteomics approach. Extracted host proteins were analyzed using two-dimensional gel electrophoresis (2D-GE) and differentially regulated proteins were identified using MALDI TOF/TOF/MS analysis. Of the 150 regulated proteins identified, 95 were downregulated while 55 were upregulated. The interaction network of regulated proteins was predicted using the STRING tool. Most downregulated proteins were involved in muscle contraction, locomotion, energy hydrolysis, lipid synthesis, serine/threonine kinase activity, oxidoreductase activity, and protein unfolding. Upregulated proteins were involved in oxidative stress pathways. Hence, cellular stress generated by S. Typhi proteins in the model host was determined using lipid peroxidation as well as oxidant and antioxidant assays. In addition, candidate proteins identified via extract analysis were validated by western blotting, and the roles of several crucial molecules were analyzed in vivo using transgenic strains (myo-2 and col-19) and mutant (ogt-1) of C. elegans. To the best of our knowledge, this is the first study to report protein regulation in host C. elegans exposed to toxic S. Typhi proteins. It highlights the significance of p38 MAPK and JNK immune pathways.

Survival upon Staphylococcus aureus mediated wound infection in Caenorhabditis elegans and the mechanism entailed.

Infection following injury is one of the major threats which causes huge economic burden in wound care management all over the world. Injury often results with poor healing when coupled by following infection. In contrast to this, we observed enhanced survival of wound infected worms compared to wounded worms in Caenorhabditis elegans wound model while infecting with Staphylococcus aureus. Hence, the study was intended to identify the mechanism for the enhanced survival of wound infected worms through LCMS/MS based high throughput proteomic analysis. Bioinformatics analyses of the identified protein players indicated differential enrichment of several pathways including MAPK signaling, oxidative phosphorylation and phosphatidylinositol signaling. Inhibition of oxidative phosphorylation and phosphatidylinositol signaling through chemical treatment showed complete reversal of the enhanced survival during wound infection nevertheless mutant of MAPK pathway did not reverse the same. Consequently, it was delineated that oxidative phosphorylation and phosphatidylinositol signaling are crucial for the survival. In this regard, elevated calcium signals and ROS including O- and H2O2 were observed in wounded and wound infected worms. Consequently, it was insinuated that presence of pathogen stress could have incited survival in wound infected worms with the aid of elevated ROS and calcium signals.

Vitexin prevents Aß proteotoxicity in transgenic Caenorhabditis elegans model of Alzheimer's disease by modulating unfolded protein response.

Alzheimer's disease (AD) accounts for an estimated 60% to 80% of all dementia cases. The present study is aimed at evaluating the neuroprotective efficacy of vitexin, an apigenin flavone glycoside using transgenic Caenorhabditis elegans strain (CL2006) of AD. The neuroprotective effect of vitexin was determined using physiological assays, quantitative polymerase chain reaction, and Western blotting. The results of survival and paralysis assay indicate that vitexin (200 µM) significantly extended the lifespan of the nematodes. Vitexin-treated nematodes showed a significant reduction in the expression of Aß, ace-1, and ace-2 genes when compared to control. Further, vitexin significantly upregulated the expression of acr-8 and dnj-14, and increased the lifespan of the nematodes. Vitexin was also found to modulate the unfolded protein response genes (hsp-4, pek-1, ire-1, and xbp-1) and suppress the expression of Aß. Overall, the results show that vitexin acts as a neuroprotective agent and protects transgenic C. elegans strains from Aß proteotoxicity.

A Paradigm Gap in Host-Pathogen Interaction Studies: Lesson from the COVID-19 Pandemic.

INTRODUCTION: COVID-19 outbreak displayed presumably an increased accumulation of SARS-CoV-2 virus during comorbid complications and a substantial variation in the mortality within and between the countries, which in turn siren us the lack of knowledge in host-pathogen interactions (HPIs). Our aim is to describe the lessons taught by the COVID-19 pandemic in the existing/missing investigations on HPI. METHODS: This was from a retrospective meta-analysis of literature on "COVID-19 and comorbidity" to expose the existing paradigm gap in HPI by highlighting the omitted concepts/areas of research and new approaches to consider for the development of future therapeutics. RESULTS: Literature on "COVID-19 and comorbidity" apparently depicted the disparity in HPI during comorbid/immune-challenged conditions, which was reflected in the poor prognosis of the disease and failed therapeutics upon clinical trials. Moreover, the entry, adherence, multiplication, and the following establishment of infection were also varied in groups with various comorbidities. This edified that the mode of interaction of an infectious agent could vary according to the immunological and health status of the host system and hence the efficiency/success rate of treatment modalities. In addition, limited number of literature on HPI upon comorbid and immune-challenged conditions of the host manifestly indicated that there is a lack of our focus/attention on consideration of the host immune/health-specific factors in HPI studies. These alert us that the development of unambiguous therapeutic approaches is needed for a better/successful treatment of novel infectious agents in the future. CONCLUSION: By understanding the immunological state exhibited in SARS-CoV-2 infection, we conclude that the COVID-19 pandemic has taught us a great lesson that our current understanding of HPI is insufficient to fight and conquer novel infections in real life. Hence, newer approaches are obligatory to understand HPI in order to combat COVID-19-like outbreaks in the future, if any, and also to design novel immunogenic/nutraceutical-based therapeutics.

Modulation of the host cell mitochondrial proteome by PemKSa toxin protein exposure.

Mitochondria are essential organelles involved in abundant cellular functions ranging from energy metabolism to cell survival. The inhibition of these mitochondrial functions by bacterial toxin proteins promotes disease and inhibits cell growth. Prominent evidence proposes that mitochondria provide a platform for innate immune response signalling pathways. To investigate how a bacterial toxin manipulates the mitochondrial environment of the host Caenorhabditis elegans at the molecular level, a quantitative proteomic study of mitochondria following exposure to the PemKSa toxin was performed. In this study, we purified C. elegans mitochondria and performed a comprehensive proteomic analysis using a shotgun proteomic approach (LC-MS/MS). LC-MS/MS data were analysed using various bioinformatics tools, which revealed the role and involvement of several regulatory proteins and pathways associated with mitochondrial functions. We detected variation in protein expression in key metabolic pathways, including oxidative phosphorylation (OXPHOS), the tricarboxylic acid (TCA) cycle, carbon metabolism, glycolysis and apoptosis, which suggests global reprogramming of host mitochondria metabolism by the toxin. Our results provide new horizons for mitochondria-associated protein functions and the classification of mitochondrial diseases during host-toxin interactions.

Lactobacillus gastricus BTM 7 prevents intestinal colonization by biofilm forming Cronobacter sakazakii in Caenorhabditis elegans model host.

The study reports protective role of potential probiotic cultures against infection by biofilm forming Cronobacter sakazakii in Caenorhabditis elegans model system. Among the fifteen indigenous potential probiotics, the cell free supernatant of Lactobacillus gastricus BTM7 possessed highest antimicrobial action and biofilm inhibition against C. sakazakii. The competitive exclusion assays revealed that preconditioning with probiotics resulted in increased mean life span of the nematode to 12-13 days as compared to 5-6 days when the pathogen was administered alone. Enhanced expression of the marker genes (pmk-1, daf-16 and skn-1) was observed during the administration of probiotic cultures. The highest expression of pmk-1 (2.5 folds) was observed with administration of L. gastricus BTM7. The principal component analysis on selected variables revealed that L. gastricus BTM7 has the potential to limit the infection of C. sakazakii in C. elegans and enhance the expression of key genes involved in extending life span of the worm.

Probiotic mediated colonization resistance against E.coli infection in experimentally challenged Caenorhabditis elegans.

This study investigates the antimicrobial potential of sixteen indigenous probiotic bacteria in Caenorhabditis elegans infected with different serotypes of clinical Escherichia coli isolates. The probiotic cultures exhibited varying degree of antimicrobial activity against the pathogenic strains. The Cell Free Supernatant (CFS) of Lactobacillus plantarum K90 exhibited maximum antimicrobial activity against all indicator strains. Further, the pathogenic potential of the clinical strains was determined using liquid killing assay in C. elegans, where the pathogenic strains resulted in complete killing of the worm in 5 days as compared to 60% survival of worms fed with standard food of E. coli OP50. The clinical strains also resulted in impaired pharynx and internal hatching of the eggs in the worms. The protective effects of probiotics against the pathogenic strains was determined via competition, exclusion and displacement assays with different stages of intervention of probiotic culture. No significant increase in mean life span (MLS) of the worm was observed in competition and displacement assay. Among the tested strains in exclusion assay, a pretreatment with L. gastricus BTM 7 was found to result in better protection of the worm against infection with pathogenic E. coli strains by extending its life by three days and no other adverse effect on physiology and morphology of the worm. The results suggest that preconditioning with probiotic strains can be used as an effective way to reduce the invasion and colonization by the pathogens.

Amyloid-ß induced neuropathological actions are suppressed by Padina gymnospora (Phaeophyceae) and its active constituent α-bisabolol in Neuro2a cells and transgenic Caenorhabditis elegans Alzheimer's model.

The inhibition of Aß peptide development and aggregation is a hopeful curative approach for the discovery of disease modifying drugs for Alzheimer's disease (AD) treatment. Recent research mainly focuses on the discovery of drugs from marine setting due to their immense therapeutic potential. The present study aims to evaluate the brown macroalga Padina gymnospora and its active constituent α-bisabolol against Aß25-35 induced neurotoxicity in Neuro2a cells and transgenic Caenorhabditis elegans (CL2006 and CL4176). The results of the in vitro study revealed that the acetone extract of P. gymnospora (ACTPG) and its active constituent α-bisabolol restores the Aß25-35 induced alteration in the oxidation of intracellular protein and lipids. In addition, ACTPG and α-bisabolol inhibited cholinesterase and ß-secretase activity in Neuro2a cells. Moreover, the intracellular reactive oxygen species (ROS) and reactive nitrogen species (RNS) production was reduced by ACTPG and α-bisabolol in Neuro2a cells. The decrease in the expression level of apoptotic proteins such as Bax and caspase-3 in ACTPG and α-bisabolol treated group indicates that the seaweed and its bioactive compound have anti-apoptotic property. Further, the in vivo study revealed that the ACTPG and α-bisabolol exerts neuroprotective effect against Aß induced proteotoxicity in transgenic C. elegans strains of AD. Moreover it altered the Aß mediated pathways, lifespan, macromolecular damage and down regulated the AD related gene expression of ace-1, hsp-4 and Aß, thereby preventing Aß synthesis. Overall, the outcome of the study signifies the neuroprotective effect of ACTPG and α-bisabolol against Aß mediated AD pathology.

In vitro and in vivo efficacy of Caenorhabditis elegans recombinant antimicrobial protein against Gram-negative bacteria.

Antimicrobial peptides (AMPs) are short, positively charged host defense peptides, found in various life forms from microorganisms to humans. AMPs are gaining more attention as substitutes for antibiotics in order to combat the risk posed by multi-drug- resistant pathogens. The nematode Caenorhabditis elegans relies solely on its innate immune defense to cope with its challenging life-style. Bacterial infection in C. elegans leads to induction of antimicrobial proteins, defensins, nemapores, cecropins, and neuropeptide-like proteins, which act to limit bacterial proliferation. This study reports how the C. elegans recombinant antibacterial factor (ABF-1) rapidly inhibited bacterial growth (Salmonella Typhi, Klebsiella pneumonia, Shigella sonnei and Vibrio alginolyticus). The ABF-1 exposure on S. Typhi, showed differential regulation in cell-cycle, DNA repair mechanism, membrane stability, and stress related proteins. The exogenous supply of ABF-1 protein has extended C. elegans survival by reducing the bacterial colony forming units on the nematode intestine. Together, these findings indicate the valuable and potential therapeutic applications of ABF-1 protein as antimicrobial agents against intracellular pathogens.

Green synthesized silver nanoparticles demonstrating enhanced in vitro and in vivo antibiofilm activity against Candida spp.

Candida species are opportunistic fungal pathogens, which are known for their biofilm associated infections on implanted medical devices in clinical settings. Broad spectrum usage of azole groups and other antifungal agents leads to the occurrence of drug resistance among Candida species. Most of the antifungal agents have failed to treat the biofilm mediated Candida infections. In the present study, silver nanoparticles (AgNPs) were synthesized using Dodonaea viscosa and Hyptis suoveolens methanolic leaf extracts and characterized by ultraviolet-visible absorption spectroscopy, X-ray diffraction analysis, Fourier transform infrared spectroscopy and Scanning electron microscopy, Dynamic light scattering, and Zeta potential analysis. The main goal of this study was to assess the AgNPs for their antibiofilm efficacy against Candida spp. through microscopic analysis and in vitro virulence assays. The results revealed that AgNPs strongly inhibited more than 80% biofilm formed by Candida spp. Furthermore, the AgNPs also reduced the yeast-to-hyphal transition, exopolysaccharide biosynthesis, secreted aspartyl proteinase production which are the major virulence factors of Candida species. This study reveals that biosynthesized AgNPs can be considered for the treatment of biofilm related Candida infections.

A combination of ellagic acid and tetracycline inhibits biofilm formation and the associated virulence of Propionibacterium acnes in vitro and in vivo.

Propionibacterium acnes is an opportunistic pathogen which has become notorious owing to its ability to form a recalcitrant biofilm and to develop drug resistance. The current study aimed to develop anti-biofilm treatments against clinical isolates of P. acnes under in vitro and in vivo conditions. A combination of ellagic acid and tetracycline (ETC; 250 µg ml(-1) + 0.312 µg ml(-1)) was determined to effectively inhibit biofilm formation by P. acnes (80-91%) without affecting its growth, therefore potentially limiting the possibility of the bacterium attaining resistance. In addition, ETC reduced the production of extracellular polymeric substances (EPS) (20-26%), thereby making P. acnes more susceptible to the human immune system and antibiotics. The anti-biofilm potential of ETC was further substantiated under in vivo conditions using Caenorhabditis elegans. This study reports a novel anti-biofilm combination that could be developed as an ideal therapeutic agent with broad cosmeceutical and pharmaceutical applicability in the era of antibiotic resistance.

An in silico, in vitro and in vivo investigation of indole-3-carboxaldehyde identified from the seawater bacterium Marinomonas sp. as an anti-biofilm agent against Vibrio cholerae O1.

Biofilm formation is a major contributing factor in the pathogenesis of Vibrio cholerae O1 (VCO1) and therefore preventing biofilm formation could be an effective alternative strategy for controlling cholera. The present study was designed to explore seawater bacteria as a source of anti-biofilm agents against VCO1. Indole-3-carboxaldehyde (I3C) was identified as an active principle component in Marinomonas sp., which efficiently inhibited biofilm formation by VCO1 without any selection pressure. Furthermore, I3C applications also resulted in considerable collapsing of preformed pellicles. Real-time PCR studies revealed the down-regulation of virulence gene expression by modulation of the quorum-sensing pathway and enhancement of protease production, which was further confirmed by phenotypic assays. Furthermore, I3C increased the survival rate of Caenorhabditis elegans when infected with VCO1 by significantly reducing in vivo biofilm formation, which was corroborated by a survivability assay. Thus, this study revealed, for the first time, the potential of I3C as an anti-biofilm agent against VCO1.

Alterations in Caenorhabditis elegans and Cronobacter sakazakii lipopolysaccharide during interaction.

Lipopolysaccharide is one of the pathogen-associated molecular patterns of Gram-negative bacteria which are essential for its pathogenicity. Cronobacter sakazakii is an opportunistic, emergent pathogen, which infects and cause mortality in Caenorhabditis elegans. In this study, modifications in host and C. sakazakii LPS during infections were evaluated. The physiological assays revealed that LPS alone is sufficient to affect the host pharyngeal pumping rate, brood size and cause lethality. FTIR spectra of LPS revealed that C. sakazakii modifies its LPS to escape from the recognition of host immune system. These results indicate that LPS plays a key role in C. sakazakii pathogenicity. qPCR studies revealed that LPS modulated the expression of selected host immune (clec-60, clec-87, lys-7, ilys-3, F08G5.6, atf-7, scl-2, cpr-2) and aging-related genes (skn-1, clk-2, bra-2, age-1, bec-1, daf-16, daf-2). Moreover, it was confirmed that p38 MAPK pathway has a major role in host immune response against LPS-mediated challenges.

Modulation of Caenorhabditis elegans immune response and modification of Shigella endotoxin upon interaction.

To analyze the pathogenesis at both physiological and molecular level using the model organism, Caenorhabditis elegans at different developmental stages in response to Shigella spp. and its pathogen associated molecular patterns such as lipopolysaccharide. The solid plate and liquid culture-based infection assays revealed that Shigella spp. infects C. elegans and had an impact on the brood size and pharyngeal pumping rate. LPS of Shigella spp. was toxic to C. elegans. qPCR analysis revealed that host innate immune genes have been modulated upon Shigella spp. infections and its LPS challenges. Non-destructive analysis was performed to kinetically assess the alterations in LPS during interaction of Shigella spp. with C. elegans. The modulation of innate immune genes attributed the surrendering of host immune system to Shigella spp. by favoring the infection. LPS appeared to have a major role in Shigella-mediated pathogenesis and Shigella employs a tactic behavior of modifying its LPS content to escape from the recognition of host immune system.

Proteomic investigation of Vibrio alginolyticus challenged Caenorhabditis elegans revealed regulation of cellular homeostasis proteins and their role in supporting innate immune system.

Caenorhabditis elegans has been the preferred model system for many investigators to study pathogenesis. In the present investigation, regulation of C. elegans proteome was explored against V. alginolyticus infection using quantitative proteomics approach. Proteins were separated using 2D-DIGE and the differentially regulated proteins were identified using PMF and MALDI TOF/TOF analysis. The results thus obtained were validated using Western blotting for candidate proteins. The corresponding transcriptional regulation was quantified subsequently using real-time PCR. Interaction network for candidate proteins was predicted using search tool for the retrieval of interacting genes/proteins (STRING) and functional validation was performed using respective mutant strains. Out of the 25 proteins identified, 21 proteins appeared to be upregulated while four were downregulated. Upregulated proteins included those involved in stress-response (PDI-2, HSP-6), immune-response (protein kinase -18, GST-8) and energy-production (ATP-2) while proteins involved in structural maintenance (IFB-2) and lipid metabolism (SODH-1) were downregulated. The roles of these players in the host system during Vibrio infection was analyzed in vivo using wild type and mutant C. elegans. Survival assays using mutants lacking pdi-2, ire-1, and xbp-1 displayed enhanced susceptibility to V. alginolyticus. Cellular stress generated by V. alginolyticus was determined using ROS assay. This is the first report of proteome changes in C. elegans against V. alginolyticus challenge and highlights the significance of unfolded protein response (UPR) pathway during bacterial infection.

Physiological and Immunological Regulations in Caenorhabditis elegans Infected with Salmonella enterica serovar Typhi.

Studies pertaining to Salmonella enterica serovar Typhimurium infection by utilizing model systems failed to mimic the essential aspects of immunity induced by Salmonella enterica serovar Typhi, as the determinants of innate immunity are distinct. The present study investigated the physiological and innate immune responses of S. Typhi infected Caenorhabditis elegans and also explored the Ty21a mediated immune enhancement in C. elegans. Ty21a is a known live vaccine for typhoidal infection in human beings. Physiological responses of C. elegans infected with S. Typhi assessed by survival and behavioral assays revealed that S. Typhi caused host mortality by persistent infection. However, Ty21a exposure to C. elegans was not harmful. Ty21a pre-exposed C. elegans, exhibited significant resistance against S. Typhi infection. Elevated accumulation of S. Typhi inside the infected host was observed when compared to Ty21a exposures. Transcript analysis of candidate innate immune gene (clec-60, clec-87, lys-7, ilys-3, scl-2, cpr-2, F08G5.6, atf-7, age-1, bec-1 and daf-16) regulations in the host during S. Typhi infection have been assessed through qPCR analysis to understand the activation of immune signaling pathways during S. Typhi infections. Gene silencing approaches confirmed that clec-60 and clec-87 has a major role in the defense system of C. elegans during S. Typhi infection. In conclusion, the study revealed that preconditioning of host with Ty21a protects against subsequent S. Typhi infection.

Cronobacter sakazakii infection implicates multifaceted neuro-immune regulatory pathways of Caenorhabditis elegans.

The neural pathways of Caenorhabditis elegans play a crucial role in regulating host immunity and inflammation during pathogenic infections. To understand the major neuro-immune signaling pathways, this study aimed to identify the key regulatory proteins in the host C. elegans during C. sakazakii infection. We used high-throughput label-free quantitative proteomics and identified 69 differentially expressed proteins. KEGG analysis revealed that C. sakazakii elicited host immune signaling cascades primarily including mTOR signaling, axon regeneration, metabolic pathways (let-363 and acox-1.4), calcium signaling (mlck-1), and longevity regulating pathways (ddl-2), respectively. The abrogation in functional loss of mTOR-associated players deciphered that C. sakazakii infection negatively regulated the lifespan of mutant worms (akt-1, let-363 and dlk-1), including physiological aberrations, such as reduced pharyngeal pumping and egg production. Additionally, the candidate pathway proteins were validated by transcriptional profiling of their corresponding genes. Furthermore, immunoblotting showed the downregulation of mTORC2/SGK-1 during the later hours of pathogen exposure. Overall, our findings profoundly provide an understanding of the specificity of proteome imbalance in affecting neuro-immune regulations during C. sakazakii infection.