Study Of Host-pathogen Interactions Research Articles

Changes in Caenorhabditis elegans immunity and Staphylococcal virulence factors during their interactions

The nematode Caenorhabditis elegans is used as a model system for the study of host-pathogen interactions. Lipoteichoic acid (LTA) is one of the major virulent and immunostimulatory components found in gram positive bacteria. The current study used LTA isolated from Staphylococcus aureus and pathogenic and non-pathogenic Staphylococcus epidermidis. The overall physiological assays revealed that LTA exposed C. elegans show a significant reduction in the life span, production of eggs and progenies. To understand the involvement of innate immune specific players at the mRNA level, the regulation of few candidate antimicrobial genes was studied during Staphylococcal LTA exposures. qPCR analysis indicated an upregulation of antimicrobial peptides during LTA exposures. To understand the involvement of LTA and other virulent genes during infection, the regulation of LTA synthase and a few virulence genes was monitored during host exposure. The qPCR analyses indicated the upregulation of ltaS and other virulence genes (atoxin, sak, ssaA and fbe) during infection. Ability of the pathogens to modify their internal machinery during host presence was monitored by Fourier transform infrared spectroscopy, electrochemical impedance spectroscopy and cyclic voltametric analyses. The FTIR results indicated distinct alterations of peaks from Staphylococcal LTA composition between control and the host exposed. Further, EIS and CV data displayed clear differences between the host exposed Staphylococcal samples compared to their respective unexposed controls. The pathogenic and non-pathogenic strains showed different types of regulations and interactions during host exposures. The observed modifications clearly suggest that the Gram positive pathogen changes its LTA production and possibly the structure to cause a severe pathogenic effect on an interacting host.

Reactivation of dormant “non-culturable” Mycobacterium tuberculosis developed in vitro after injection in mice: Both the dormancy depth and host genetics influence the outcome

Three stocks of Mycobacterium tuberculosis H37Rv were cultured in vitro under prolonged hypoxic or acidified conditions until partial or complete loss of the capacity to form colonies on agar medium was achieved. Such dormant “non-culturable” mycobacteria were assessed for the growth resuscitation after intra-tracheal injection into mice of the two inbred strains with different genetic susceptibility to M. tuberculosis-triggered disease: hyper-susceptible I/St and relatively resistant B6. The results indicate that bacteria which are able to resuscitate spontaneously in liquid medium in vitro started to multiply in organs of infected mice, and that the outcome of such infection strongly depended upon the level of genetic TB susceptibility. However, dormant bacteria required inducers for resuscitation in vitro lost the capacity to multiply even in genetically susceptible mice. The established model of dormancy/reactivation is suitable for the studying host–pathogen interactions and testing vaccine and drug candidates specifically targeting latent TB.

A Comparison of the Chicken and Turkey Proteomes and Phosphoproteomes in the Development of Poultry-Specific Immuno-Metabolism Kinome Peptide Arrays.

The use of species-specific peptide arrays for the study of animal kinomes has a proven track record of success. This technique has been used in a variety of species for the study of host–pathogen interactions and metabolism. Species-specific peptide arrays have been designed previously for use with chicken but a turkey array has never been attempted. In addition, arrays designed around individual cellular functions have been designed and utilized, but cross-function immuno-metabolic arrays have not been considered previously. Antecedent to designing separate chicken and turkey immuno-metabolic kinome peptide arrays, we show that while the chicken and turkey genomes are quite similar, the two species are much more distinct at the proteome and phosphoproteome levels. Despite a genome identity of approximately 90%, we observe that only 83% of chicken and turkey orthologous proteins display sequence matches between the two species. Further, less than 70% of kinase recognition target sequences are exact matches between chicken and turkey. Thus, our analysis shows that, at the proteome and kinome level, these two species must be considered separately in the design of novel peptide arrays. Our ultimate array design covers numerous immune and metabolic processes including innate and adaptive immunity, inflammatory responses, carbohydrate, protein, and fat metabolism, and response to hormones. We have shown the proteomic and phosphoproteomic diversity of chicken and turkey and have designed a valuable research tool for the study of immuno-metabolism within these two species.

Significant Effects Due to Peptone in Kelman Medium on Colony Characteristics and Virulence of Ralstonia solanacearum in Tomato

The study was taken up to assess if the media constituents played any role in governing the variable colony characteristics or pathogenicity of the bacterial wilt pathogen, Ralstonia solanacearum cultured on the widely employed Kelman medium. The effects due to the constituents 2,3,5-triphenyl tetrazolium chloride (TTC), peptone, casein hydrolysate and glucose on colony characteristics were investigated using -80°C stored culture of strain ‘NH-Av01’ (race 1, biovar 3) isolated from tomato. Comparing the pigment inducing TTC from two brands, its source or mode of storage/incorporation did not impart any significant effects. The source of peptone, on the other hand, displayed striking effects on the extent of colony growth, fluidity and red pigmentation depending on type, brand or batch / lot of manufacture as documented with 20 different formulations. Significant differences in the pathogenicity of isolate derived from different peptone sources in seedling-challenge assay on tomato were observed. The observations on peptone effects were endorsed with four other isolates belonging to distinct geographic locations, crops (eggplant, chilli, ginger) or races (race 1 or 4). The peptone source did not influence the pathogen-responses in biovar tests but notably altered the pattern of lawn formation and inhibition zone development during antagonistic assays. Casein hydrolysate displayed some variable effects while glucose source had no effect. This study brings to light the significant modifying effects by the peptone-constituent in Kelman medium on the physiology of R. solanacearum and the virulence of isolate and the need to consider the source of media components during culture maintenance, host-pathogen interaction studies or microbe-microbe interaction investigations.

Quantitative proteome analysis of Caenorhabditis elegans upon exposure to nematicidal Bacillus thuringiensis.

This study identified system-wide effects of Bt intoxication on C. elegans at protein level, expanding the catalogue of immune effectors potentially acting towards the pathogen, and provide verification for numerous gene products implicated in previous transcriptomic studies. The data present evidence in support of both a general defense response as well as a specific reaction against the Bt toxin within the nematode. The described findings will also contribute to a deeper understanding of host-microbe interaction in other organisms, including humans, and may provide key information that touches far reaching aspects of coevolutionary processes.

Next-generation sequencing in veterinary medicine: how can the massive amount of information arising from high-throughput technologies improve diagnosis, control, and management of infectious diseases?

The development of high-throughput molecular technologies and associated bioinformatics has dramatically changed the capacities of scientists to produce, handle, and analyze large amounts of genomic, transcriptomic, and proteomic data. A clear example of this step-change is represented by the amount of DNA sequence data that can be now produced using next-generation sequencing (NGS) platforms. Similarly, recent improvements in protein and peptide separation efficiencies and highly accurate mass spectrometry have promoted the identification and quantification of proteins in a given sample. These advancements in biotechnology have increasingly been applied to the study of animal infectious diseases and are beginning to revolutionize the way that biological and evolutionary processes can be studied at the molecular level. Studies have demonstrated the value of NGS technologies for molecular characterization, ranging from metagenomic characterization of unknown pathogens or microbial communities to molecular epidemiology and evolution of viral quasispecies. Moreover, high-throughput technologies now allow detailed studies of host-pathogen interactions at the level of their genomes (genomics), transcriptomes (transcriptomics), or proteomes (proteomics). Ultimately, the interaction between pathogen and host biological networks can be questioned by analytically integrating these levels (integrative OMICS and systems biology). The application of high-throughput biotechnology platforms in these fields and their typical low-cost per information content has revolutionized the resolution with which these processes can now be studied.The aim of this chapter is to provide a current and prospective view on the opportunities and challenges associated with the application of massive parallel sequencing technologies to veterinary medicine, with particular focus on applications that have a potential impact on disease control and management.

Effects of antibiofilm compounds on the cellular and bacterial colonization of polimeric surfaces: a step towards biofunctionalization of implantable devices

Background Periodontal and peri-implant diseases are infection pathologies related to the pathogenicity of the microorganisms involved [1]. The worldwide practice of indiscriminate and continuous use of antibiotics for the control and prophylaxis of bacterial pathogens has led to the development of bacterial resistance to most available antimicrobials [2]. Concurrently, the rapid increase in bacterial resistance to currently available antimicrobial drugs has led researchers to search for new sources of molecules active against bacterial pathogens, outlining a new generation of anti-infective drug development. A promising approach for the development of a new generation of antimicrobial drugs has arisen from the studies of host-pathogen interactions, which prompted a shift of drug targets from bacterial survival to pathogenicity control. Examples of potential nonconventional targets for microbial control are molecules and receptors involved in bacterial adherence to biotic and abiotic surfaces as well as signal systems controlling bacterial group behaviour of populations organised in biofilms, such as quorum sensing (QS) [3]. In contrast to conventional antibiotics, antimicrobial drugs directed against such unconventional targets do not jeopardise bacterial survival, imposing a low selection pressure and thus avoiding the development of resistance [4]. Methods Here we analysed the effects of novel synthetic lactams, compounds derived from furanones which display recognized antibiofilm activity. Three synthetic lactams were tested against biofilm of Enterococcus faecalis grown over an implantable polymeric material (PLGA-HA). Inhibition assays of adhesion and biofilm formation, and SEM observation were used to quantify and qualify E. faecalis’ biofilm formation. To assay for biofilm inhibition, bacterial strains were grown statically for 20 h at 37°C in TSB. Polystyrene 96-well microtitre plates were inoculated with 100 μL/well of bacterial suspension previously diluted to 5 × 10 CFU/mL in TSB supplemented with 4% sucrose (w/v) and 3.5% (v/v) dimethyl sulphoxide (DMSO). Compounds to be tested for antibiofilm activity were prepared at a concentration of 175 μg/ml, in the same media as before, and added to the test wells. Biofilm inhibition capacity was assessed by CFU counting after bacterial disaggregation from the surfaces and microscopic analysis of the polymers. Cytotoxicity of the lactams against human fibroblasts and keratinocytes was tested, at four different concentrations (43,75; 87,5; 131,25 and 175 μg/ml), by MTT assay [5].

Nachweis von Bakterien, Pilzen und Viren im Hochdurchsatz

Next generation sequencing (NGS) not only provides a powerful tool for genome-wide studies of host-pathogen interactions, but also allows the unbiased detection of known as well as novel pathogens of fungal, bacterial and viral origin in clinical samples. Given this, NGS-based diagnostic approaches have a unique potential to fundamentally improve the level of preparedness for infectious disease outbreaks. Here, we describe the advantages and limitations of current approaches for the identification of pathogens by NGS.

A small intestinal organoid model of non-invasive enteric pathogen-epithelial cell interactions.

Organoids mirror in vivo tissue organization and are powerful tools to investigate the development and cell biology of the small intestine. However, their application for the study of host-pathogen interactions has been largely unexplored. We have established a model using microinjection of organoids to mimic enteric infection, allowing for direct examination of pathogen interactions with primary epithelial cells in the absence of confounding variables introduced by immune cells or the commensal microbiota. We investigated the impact of Paneth cell α-defensin antimicrobial peptides on bacterial growth. We demonstrate that organoids form a sealed lumen which contains concentrations of α-defensins capable of restricting growth of multiple strains of Salmonella enterica serovar Typhimurium for at least 20 h post-infection. Transgenic expression of human defensin 5 (HD5) in mouse organoids lacking functional murine α-defensins partially restored bacterial killing. We also found that organoids from NOD2−/− mice were not impaired in α-defensin expression or antibacterial activity. This model is optimized for the study of non-invasive bacteria, but can be extended to other enteric pathogens and is amenable to further genetic manipulation of both the host and microbe to dissect this critical interface of host defense.

Amoebae as a tool to isolate new bacterial species, to discover new virulence factors and to study the host–pathogen interactions

Amoebae are unicellular protozoan present worldwide in several environments mainly feeding on bacteria. Some of them, the amoebae-resistant bacteria (ARBs), have evolved mechanisms to survive and replicate inside amoebal species. These mainly include legionella, mycobacteria and Chlamydia-related bacteria. Amoebae can provide a replicative niche, can act as reservoir for bacteria whereas the cystic form can protect the internalized bacteria. Moreover, the amoebae represent a Trojan horse for ARBs to infect animals. The long interaction between amoebae and bacteria has likely selected for bacterial virulence traits leading to the adaptation towards an intracellular lifestyle, and some ARBs have acquired the ability to infect mammals.This review intends to highlight the important uses of amoebae in several fields in microbiology by describing the main tools developed using amoebal cells. First, amoebae such as Acanthamoeba are used to isolate and discover new intracellular bacterial species by two main techniques: the amoebal co-culture and the amoebal enrichment. In the second part, taking Waddlia chondrophila as example, we summarize some important recent applications of amoebae to discover new bacterial virulence factors, in particular thanks to the amoebal plaque assay. Finally, the genetically tractable Dictyostelium discoideum is used as a model organism to study host–pathogen interactions, in particular with the development of several approaches to manipulate its genome that allowed the creation of a wide range of mutated strains largely shared within the Dictyostelium community.

Editorial (Thematic Issue: Current Status and Perspective on Drug Targets in Tubercle Bacilli and Drug Design of Antituberculous Agents Based on Structure-Activity Relationship)

Editorial (Thematic Issue: Current Status and Perspective on Drug Targets in Tubercle Bacilli and Drug Design of Antituberculous Agents Based on Structure-Activity Relationship)

The utilization of oropharyngeal intratracheal PAMP administration and bronchoalveolar lavage to evaluate the host immune response in mice.

The host immune response to pathogens is a complex biological process. The majority of in vivo studies classically employed to characterize host-pathogen interactions take advantage of intraperitoneal injections of select bacteria or pathogen associated molecular patterns (PAMPs) in mice. While these techniques have yielded tremendous data associated with infectious disease pathobiology, intraperitoneal injection models are not always appropriate for host-pathogen interaction studies in the lung. Utilizing an acute lung inflammation model in mice, it is possible to conduct a high resolution analysis of the host innate immune response utilizing lipopolysaccharide (LPS). Here, we describe the methods to administer LPS using nonsurgical oropharyngeal intratracheal administration, monitor clinical parameters associated with disease pathogenesis, and utilize bronchoalveolar lavage fluid to evaluate the host immune response. The techniques that are described are widely applicable for studying the host innate immune response to a diverse range of PAMPs and pathogens. Likewise, with minor modifications, these techniques can also be applied in studies evaluating allergic airway inflammation and in pharmacological applications.

Agrobacterium tumefaciens-mediated transformation for targeted disruption and over expression of genes in the poplar pathogenSphaerulina musiva

Summary Sphaerulina musiva causes both leaf spots and cankers on poplar. Leaf spots can lead to defoliation and cankers on branches and primary stems can lead to stem breakage and tree mortality. The recent availability of both the S. musiva and Populus trichocarpa genomes offers a great opportunity to study host–pathogen interactions. To better understand the factors involved in S. musiva pathology, we present a strategy for the transformation of this species using Agrobacterium tumefaciens. Binary plasmids were generated with hygromycin B phosphotransferase (hph) flanked by upstream and downstream sequences of polyketide synthase-like (PKS-L1) gene to generate targeted gene disruptants by homologous recombination. Plasmids were also constructed for constitutive expression reporter genes eGFP and mCherry to help with histological characterization of the pathogen during infection. Gene knockouts were identified by PCR and confirmed by sequencing and Southern blotting. No significant differences were observed in melanin production between PKS-L1 disruptants and wild type isolates. Colonies expressing reporter genes were identified by fluorescent stereomicroscopy. This method is a promising tool for the characterization of pathogen genes through reverse and forward genetics and for introducing markers for histopathological study.

Intravital two-photon imaging to understand bacterial infections of the mammalian host.

Intravital two-photon microscopy (2PM) is an advanced fluorescence based imaging technique that allows for a cinematic study of physiological events occurring within tissues of the live animal. Based on this real-time imaging platform, the pathophysiology of bacterial infections can be studied in the most relevant of model systems-the live host. Whereas traditional animal models of host-pathogen interaction studies rely on end stage analysis of dissected tissues, noninvasive intravital imaging allows for real-time monitoring of infection during shorter or extended time frames. Here we describe the use of advanced surgical techniques for initiation of spatially and temporally well-controlled kidney infections in rats, and how the bacterial whereabouts can be studied while at the same time monitoring the host's altered tissue homeostasis based on real-time deep tissue imaging on the 2PM platform. Whereas this chapter focuses on pyelonephritis induced by uropathogenic Escherichia coli (UPEC) in rats, the major concepts can easily be translated to numerous infections in a variety of organs.

Can Mycobacterial Genomics Generate Novel Targets as Speed-Breakers Against the Race for Drug Resistance

The rise of multi-drug resistant and extensively drug resistant M. tuberculosis around the world poses a great threat to human health, and necessitates development of new, effective and inexpensive anti-tubercular agents. The availability of knowledge on molecular biology of M. tuberculosis infection coupled with whole genome sequences, transcriptomic, proteomic and metabolomic data sets have provided insights on the genes/proteins indispensable for initiation and maintenance of persistence, cross-talk with and/or sensing the host immune response, and finally the reactivation of persistent M. tuberculosis to a growing state. The review will focus on analysis of current state of M. tuberculosis genomic resources, host-pathogen interaction studies in the context of pathogen persistence, and the efforts made or required in the development and utilization of computational tools, models and metabolic network analyses to speed up the process of drug target discovery, particularly eradicating the dormant infections.

Host Adaptation Is Contingent upon the Infection Route Taken by Pathogens

Evolution of pathogen virulence is affected by the route of infection. Also, alternate infection routes trigger different physiological responses on hosts, impinging on host adaptation and on its interaction with pathogens. Yet, how route of infection may shape adaptation to pathogens has not received much attention at the experimental level. We addressed this question through the experimental evolution of an outbred Drosophila melanogaster population infected by two different routes (oral and systemic) with Pseudomonas entomophila. The two selection regimes led to markedly different evolutionary trajectories. Adaptation to infection through one route did not protect from infection through the alternate route, indicating distinct genetic bases. Finally, relatively to the control population, evolved flies were not more resistant to bacteria other than Pseudomonas and showed higher susceptibility to viral infections. These specificities and trade-offs may contribute to the maintenance of genetic variation for resistance in natural populations. Our data shows that the infection route affects host adaptation and thus, must be considered in studies of host-pathogen interaction.

The ovarian transcriptome of the cattle tick, Rhipicephalus (Boophilus) microplus, feeding upon a bovine host infected with Babesia bovis

BackgroundCattle babesiosis is a tick-borne disease of cattle with the most severe form of the disease caused by the apicomplexan, Babesia bovis. Babesiosis is transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus. The most prevalent species is Rhipicephalus (Boophilus) microplus, which is distributed throughout the tropical and subtropical countries of the world. The transmission of B. bovis is transovarian and a previous study of the R. microplus ovarian proteome identified several R. microplus proteins that were differentially expressed in response to infection. Through various approaches, we studied the reaction of the R. microplus ovarian transcriptome in response to infection by B. bovis.MethodsA group of ticks were allowed to feed on a B. bovis-infected splenectomized calf while a second group fed on an uninfected splenectomized control calf. RNA was purified from dissected adult female ovaries of both infected and uninfected ticks and a subtracted B. bovis-infected cDNA library was synthesized, subtracting with the uninfected ovarian RNA. Four thousand ESTs were sequenced from the ovary subtracted library and annotated.ResultsThe subtracted library dataset assembled into 727 unique contigs and 2,161 singletons for a total of 2,888 unigenes, Microarray experiments designed to detect B. bovis-induced gene expression changes indicated at least 15 transcripts were expressed at a higher level in ovaries from ticks feeding upon the B. bovis-infected calf as compared with ovaries from ticks feeding on an uninfected calf. We did not detect any transcripts from these microarray experiments that were expressed at a lower level in the infected ovaries compared with the uninfected ovaries. Using the technique called serial analysis of gene expression, 41 ovarian transcripts from infected ticks were differentially expressed when compared with transcripts of controls.ConclusionCollectively, our experimental approaches provide the first comprehensive profile of the R. microplus ovarian transcriptome responding to infection by B. bovis. This dataset should prove useful in molecular studies of host-pathogen interactions between this tick and its apicomplexan parasite.

Gastrointestinal Cell Lines Form Polarized Epithelia with an Adherent Mucus Layer when Cultured in Semi-Wet Interfaces with Mechanical Stimulation

Mucin glycoproteins are secreted in large quantities by mucosal epithelia and cell surface mucins are a prominent feature of the glycocalyx of all mucosal epithelia. Currently, studies investigating the gastrointestinal mucosal barrier use either animal experiments or non-in vivo like cell cultures. Many pathogens cause different pathology in mice compared to humans and the in vitro cell cultures used are suboptimal because they are very different from an in vivo mucosal surface, are often not polarized, lack important components of the glycocalyx, and often lack the mucus layer. Although gastrointestinal cell lines exist that produce mucins or polarize, human cell line models that reproducibly create the combination of a polarized epithelial cell layer, functional tight junctions and an adherent mucus layer have been missing until now. We trialed a range of treatments to induce polarization, 3D-organization, tight junctions, mucin production, mucus secretion, and formation of an adherent mucus layer that can be carried out using standard equipment. These treatments were tested on cell lines of intestinal (Caco-2, LS513, HT29, T84, LS174T, HT29 MTX-P8 and HT29 MTX-E12) and gastric (MKN7, MKN45, AGS, NCI-N87 and its hTERT Clone5 and Clone6) origins using Ussing chamber methodology and (immuno)histology. Semi-wet interface culture in combination with mechanical stimulation and DAPT caused HT29 MTX-P8, HT29 MTX-E12 and LS513 cells to polarize, form functional tight junctions, a three-dimensional architecture resembling colonic crypts, and produce an adherent mucus layer. Caco-2 and T84 cells also polarized, formed functional tight junctions and produced a thin adherent mucus layer after this treatment, but with less consistency. In conclusion, culture methods affect cell lines differently, and testing a matrix of methods vs. cell lines may be important to develop better in vitro models. The methods developed herein create in vitro mucosal surfaces suitable for studies of host-pathogen interactions at the mucosal surface.

Multitask learning for host–pathogen protein interactions

Motivation: An important aspect of infectious disease research involves understanding the differences and commonalities in the infection mechanisms underlying various diseases. Systems biology-based approaches study infectious diseases by analyzing the interactions between the host species and the pathogen organisms. This work aims to combine the knowledge from experimental studies of host–pathogen interactions in several diseases to build stronger predictive models. Our approach is based on a formalism from machine learning called ‘multitask learning’, which considers the problem of building models across tasks that are related to each other. A ‘task’ in our scenario is the set of host–pathogen protein interactions involved in one disease. To integrate interactions from several tasks (i.e. diseases), our method exploits the similarity in the infection process across the diseases. In particular, we use the biological hypothesis that similar pathogens target the same critical biological processes in the host, in defining a common structure across the tasks.Results: Our current work on host–pathogen protein interaction prediction focuses on human as the host, and four bacterial species as pathogens. The multitask learning technique we develop uses a task-based regularization approach. We find that the resulting optimization problem is a difference of convex (DC) functions. To optimize, we implement a Convex–Concave procedure-based algorithm. We compare our integrative approach to baseline methods that build models on a single host–pathogen protein interaction dataset. Our results show that our approach outperforms the baselines on the training data. We further analyze the protein interaction predictions generated by the models, and find some interesting insights.Availability: The predictions and code are available at: http://www.cs.cmu.edu/∼mkshirsa/ismb2013_paper320.htmlContact: j.klein-seetharaman@warwick.ac.ukSupplementary information: Supplementary data are available at Bioinformatics online.

Characterization of abalone Haliotis tuberculata–Vibrio harveyi interactions in gill primary cultures

The decline of European abalone Haliotis tuberculata populations has been associated with various pathogens including bacteria of the genus Vibrio. Following the summer mortality outbreaks reported in France between 1998 and 2000, Vibrio harveyi strains were isolated from moribund abalones, allowing in vivo and in vitro studies on the interactions between abalone H. tuberculata and V. harveyi. This work reports the development of primary cell cultures from abalone gill tissue, a target tissue for bacterial colonisation, and their use for in vitro study of host cell-V. harveyi interactions. Gill cells originated from four-day-old explant primary cultures were successfully sub-cultured in multi-well plates and maintained in vitro for up to 24days. Cytological parameters, cell morphology and viability were monitored over time using flow cytometry analysis and semi-quantitative assay (XTT). Then, gill cell cultures were used to investigate in vitro the interactions with V. harveyi. The effects of two bacterial strains were evaluated on gill cells: a pathogenic bacterial strain ORM4 which is responsible for abalone mortalities and LMG7890 which is a non-pathogenic strain. Cellular responses of gill cells exposed to increasing concentrations of bacteria were evaluated by measuring mitochondrial activity (XTT assay) and phenoloxidase activity, an enzyme which is strongly involved in immune response. The ability of gill cells to phagocyte GFP-tagged V. harveyi was evaluated by flow cytometry and gill cells-V. harveyi interactions were characterized using fluorescence microscopy and transmission electron microscopy. During phagocytosis process we evidenced that V. harveyi bacteria induced significant changes in gill cells metabolism and immune response. Together, the results showed that primary cell cultures from abalone gills are suitable for in vitro study of host-pathogen interactions, providing complementary assays to in vivo experiments.