does salmonella typhi produce toxins

This mutant strain is defective in the SPI-2 T3SS and consequently has an altered intracellular vacuole and is therefore unable to efficiently package typhoid toxin into vesicle transport carriers (Chang et al., 2016). Federal government websites often end in .gov or .mil. The quantification of typhoid toxin-associated fluorescent puncta, a measure of typhoid toxin carrier intermediates in infected cells, is shown in (e). Washing fresh produce with water helps remove harmful bacteria and germs, but it doesn't completely get rid of them. Scale bar = 5 m. The three most common causative organisms of bacterial dysentery are Campylobacter, Shigella, and Salmonella. Furthermore, we approached the study in a gradual manner so as to be able to use the information obtained from the examination of some candidates to select additional ones. The relative toxicity of the different samples, shown in (h), was measured by determining the percentage of cells in the G2/M phase from the results of the dilution of infection media experiments (shown in f and g) fitted by nonlinear regression. The export pathway, which has evolved to be specifically adapted to the biology of Salmonella Typhi, has coopted cellular machinery involved in various secretory and exocytic pathways (Figure 7). In the interests of transparency, eLife publishes the most substantive revision requests and the accompanying author responses. We found that in CIM6PR -/- cells, formation of the CdtB/Sec23 complex was impaired (these data are shown in a modified Figure 4). In many countries, E. coli is not as common as Salmonella in causing foodborne illness and outbreaks, primarily due to inadequate surveillance measures. You can reduce your risk of salmonella with safe food handling habits and by washing your hands after touching animals. (e) Quantification of the intensity of typhoid toxin-associated fluorescent puncta associated with typhoid toxin carrier intermediates in cells infected with S. Typhi expressing either wild-type SseJ or its catalytic mutant SseJS151A. (b) Quantification of the intensity of fluorescent puncta associated with typhoid toxin carrier intermediates in parental HEK293T and the indicated deficient cells. The toxin does the heavy lifting in the disease process, using a specific protein one-two punch technique when attacking its prey, white blood cells. Viral variants from the two pathways showed differential selection of compensatory mutations in Gag cleavage sites. This observation (shown in the revised manuscript as Supplementary Figure S17) is consistent with the involvement of COPII in the packaging and trafficking of typhoid toxin to the extracellular space. This is an example of which type of microbial source for foodborne illness? (b) The interaction between typhoid toxin and CI-M6PR was verified in Salmonella Typhi-infected cells. Salmonella infection (salmonellosis) is a common bacterial disease that affects the intestinal tract. These points could be mentioned in the 'limitations of this study' paragraph in the Discussion. Ideally, we would have preferred to identify components of the typhoid toxin export pathway through a genome-wide screen as we have done for the incoming endocytic pathway using CRISPR/Cas9 [see Chang & Galan, PLoS Pathogens (2019) PMID: 30951565]. eCollection 2021. Your revised article has been evaluated by Dominique Soldati-Favre (Senior Editor) and a Reviewing Editor. You should complement the CRISPR KO cell lines described in this study with a plasmid expressing the deleted gene to confirm that the phenotype is not due to a random mutation acquired during the process. Here we show that deletion of genes encoding the binding subunit (pltB) and a bacteriophage muramidase predicted to play a role in toxin export (ttsA) does not abolish toxin activity in the S-CDT-positive NTS Salmonella enterica subsp. We are aware that it has been reported that infection of cultured cells with Salmonella Typhimurium results in a redistribution of the CI-M6PR and the release of proteases to the infection supernatant. This observation further supports the premise that the CI-M6PR/COPII complex serves as cargo receptor for typhoid toxin (shown in the revised manuscript as Figure 4j). The mechanisms by which Rab11B may regulate typhoid toxin transport are unclear. Values are the mean SD of three independent experiments. The different cell lines were then infected with S. Typhi and the levels of typhoid toxin in the infection media, a direct measure of its export, were determined. Little is known about the de novo resistance pathway for DRV. However, we found that Rab32 and Rab38 are not required for typhoid toxin transport since cells deficient in HPS4, an essential component of their exchange factor (Gerondopoulos et al., 2012), were unaffected in typhoid toxin transport. The organism, now named Salmonella enterica serotype Typhi, was discovered in 1880. official website and that any information you provide is encrypted Lysosomal hydrolases, for example, are not recruited to the wild-type S. Typhi containing vacuole (see Supplementary Figure S1). In previous studies, Song discovered that Salmonella typhi produce a toxin that is responsible for most of the symptoms of typhoid fever. -, Parry CM, Threlfall EJ. A small number of people who are treated may feel better after treatment. Unauthorized use of these marks is strictly prohibited. n.sd: differences not statistically significant. See this image and copyright information in PMC. We have introduced the suggested editorial changes. CI-M6PR: cation independent mannose-6-phosphate receptor; TT: typhoid toxin. Values were normalized relative to parental cells, which was considered to be 100, and are the mean SEM. 2014 Dec 4;159(6):1290-9. doi: 10.1016/j.cell.2014.10.057. 2010;50(2):241246. Keywords: Epub 2013 Jul 10. As indicated by the reviewer, the absence of AP4 resulted in a reduced level of typhoid toxin export. Salmonella gastroenteritis, also called salmonellosis, is caused by the rod-shaped, gram-negative bacterium Salmonella. Salmonella Heidelberg has caused outbreaks associated with raw produce. The reviewer is correct and, as expected, the S. Typhi spiA mutant is directed to lysosomes. Surprisingly, we found that two toxins play different roles despite the high similarity between the two, she says. These results indicate that the presence of high level of free SehA toxin in Salmonella does not impact the viability of infected cells. It is well established that, as an intracellular pathogen, Salmonella builds its own vacuolar compartment shaped by the activity of its arsenal of effector proteins delivered by its two type III secretion systems (Galn, 2001; Jennings et al., 2017). However, we hypothesize that this GTPase may be involved in the movement of the typhoid toxin carriers along microtubule tracks. Neutralization of Typhoid Toxin by Alpaca-Derived, Single-Domain Antibodies Targeting the PltB and CdtB Subunits. As COPII is responsible for ER export there is a high possibility that blocking it through knockout could result in the mislocalisation of many proteins and subsequent off-target effects; this is not fully addressed. We specifically examined the potential contribution to typhoid toxin packaging of four well-characterized coat or adaptor proteins: clathrin (Briant et al., 2020), coat protein complex II (COPII) (McCaughey and Stephens, 2018), and adaptor-related protein complex 3 (AP3) and 4 (AP4) (Hirst et al., 2013; Odorizzi et al., 1998). Perhaps the reviewer missed the data, but we did compare the recruitment of CI-M6PR to the SCV of cells infected with wild-type S. Typhi and S. Typhi expressing either wild-type SseJ or its catalytic mutant SseJS151A. Cells were infected with a S. Typhi strain expressing FLAG-tagged CdtB and the levels of fluorescence associated with typhoid toxin carriers were determined 24 hr after infection. Despite the vast difference in disease outcomes that S.Typhi and S.Typhimurium cause in humans, there are few genomic regions that are unique to S.Typhi. **: p<0.01, unpaired two-sided t test. Henle-407 cells were infected with S. Typhi expressing FLAG-tagged CdtB for 24 hr and the interaction between typhoid toxin and endogenous CI-M6PR was probed by affinity purification with a FLAG antibody (directed to the CdtB subunit of typhoid toxin) and western blot (with antibodies to both FLAG and anti-CI-M6PR). Toxins (Basel). The properties of the SCVs are determined by the activity of bacterial effectors delivered by either of its type III secretion systems, which differ significantly between S. Typhi and S. Typhimurium. Consistent with the involvement of VAMP-7 in typhoid toxin export, we found reduced toxin levels in the infection media of VAMP-7-deficient cells infected with S. Typhi (Figure 6a and Figure 6source data 1), although the levels of typhoid toxin carrier intermediates were not significantly altered in these cells (Figure 6c and Figure 5figure supplement 3, and Figure 6figure supplement 2). N Engl J Med. Other foods like green vegetables, fruit. Recent genomic sequence analyses in several organisms suggest that new functions encoded by small open reading frames (sORFs) may emerge de novo from noncoding sequences. This toxin has an unusual biology in that it is produced by Salmonella Typhi only when located within host cells. Next day, the protein complex was incubated with 20 l of anti-FLAG M2 agarose for 2 hr at 4C. Relating to SseJ's role, examining the altered expression and/or localization of OSBP and CI-M6PR on the SCV in various cases tested in the manuscript (i.e., S. Typhi-, S. Typhi+sseJ-, and S. Typhi+sseJS151A-infected host cells) is expected to strengthen the authors' point on the co-evolution of SPI2 effectors and typhoid toxin. CAMPYLOBACTERIOSIS These observations are intriguing since it is well established that CI-M6PR is not recruited to the vacuoles that contain S. Typhimurium (Garcia-del Portillo and Finlay, 1995; McGourty et al., 2012), which does not encode typhoid toxin. Dynamic Duo-The Salmonella Cytolethal Distending Toxin Combines ADP-Ribosyltransferase and Nuclease Activities in a Novel Form of the Cytolethal Distending Toxin. A human volunteer study suggested that typhoid toxin does not appear to be required for the initiation of S. Typhi infection (Gibani et al., 2019). The findings, published Feb. 21 in the journal PLOS Pathogens, provide new directions for developing . eCollection 2021 May 21. Keep uncooked meats separate from produce, cooked foods, and ready-to-eat foods. Recent Advances in the Detection of Antibiotic and Multi-Drug Resistant, NCI CPTC Antibody Characterization Program, Crump JA, Luby SP, Mintz ED. . Salmonella can be found in a variety of foods, including chicken, beef, pork, eggs, fruits, vegetables, and even processed foods. However, in this compartment, as we have previously shown, the mere presence of CI-M6PR on the SCV is not sufficient for typhoid toxin packaging into vesicle carrier intermediates since packaging requires the intracellular environment of the wild-type SCV (see PMID: 27832592). All cell lines were routinely tested for a mycoplasma by a standard PCR method. Protein complexes were collected by centrifugation at 500 g for 1 min, washed three times with lysis buffer, and then resuspended in 30 l of Laemmli sample buffer. PMC Again, we thank the reviewer for the thorough review of our work. 1. Infection with wild-type S. Javiana activates a DNA damage response and results in, The presence of either artB or pltB is essential for S-CDT-mediated intoxication. The site is secure. Whenever possible, however, we have attempted to provide alternative pieces of evidence to support our conclusions. It is "the nature of the beast" Regardless, when appropriate, we have tried to place our findings in the context of previously described pathways. 2022 Feb 17;90(2):e0051521. 2014 Nov;72(2):95-103. doi: 10.1111/2049-632X.12191. The intensity of fluorescence associated with toxin carriers was normalized using the fluorescence-associated typhoid toxin within bacterial cells in the same field. Antimicrobial resistance in typhoidal and nontyphoidal salmonellae. However, the identity of such putative receptor is unknown. It is one of the more common causes of gastroenteritis with more than 2300 cases occurring in Georgia each year. In previous studies, Song discovered that Salmonella typhi produce a toxin that is responsible for most of the symptoms of typhoid fever. Bacterial Toxins Are a Never-Ending Source of Surprises: From Natural Born Killers to Negotiators. Values are the mean SD of three independent experiments. Salmonella Typhi pathogenesis; bacterial pathogenesis; bacterial toxins; host-pathogen interactions; typhoid fever. . To test this hypothesis, we individually expressed in S. Typhi the S. Typhimuirum SPI-1 or SPI-2 T3SS effector proteins that are absent or pseudogenes in S. Typhi (Parkhill et al., 2001) (i.e. Would you like email updates of new search results? 2) The co-IP experiment shown in Figure 4I could be performed in the CI-M6PR-/- background. The wild-type S. enterica serovars Typhi ISP2825 (Galn and Curtiss, 1991) and Typhimurium SL1344 (Hoiseth and Stocker, 1981) have been described previously. This hypothesis is based on the observation that Rab11B has been shown to modulate the exocytic transport of insulin granules at least in part by linking these transport carriers to kinesin motors, a function that requires the Rab11B effector Rip11 (Sugawara et al., 2009). ****: p<0.0001. Co-localization between CI-M6PR and SCV were analyzed by performing the plug-in Manders overlap coefficient in ImageJ. The relative toxin export is shown in (h) and was determined as described in (c) and (d). We therefore generated SNAP23-deficient cells by CRISPR/Cas9 gene editing (Figure 5figure supplement 1), infected them with S. Typhi, and examined the levels of typhoid-toxin carrier intermediates in the infected cells and the levels of typhoid toxin in the culture media. (f) Western blot analysis of the expression of typhoid toxin in parental HEK293T and Sar1B-deficient cells. For the purposes of this article we are going to talk about two groups of this bacteria- typhoidal (S. typhi) and non-typhoidal (salmonella spp.). Values are the mean SD of three independent experiments. From this location, the toxin is then packaged into vesicle carriers and exported to the extracellular medium, from where it finds its way into . Western blot analysis should be used to determine if lower amounts in extracellular fractions can be rescued by protease inhibition. Although cells deficient in Rab11B showed reduced amount of toxin in the infection media, they showed normal levels of vesicle carrier intermediates. The incidence of typhoid fever in sub-Saharan Africa was an estimated 725 cases/100,000 persons in 2010, despite a lack of incidence studies conducted in West and central Africa ( 1 ). Infection media obtained from S. Typhi-infected HEK293T parental and the indicated deficient cells were serially diluted as indicated and applied to uninfected HEK293T cells. (a and b) Purified FLAG-tagged wild-type typhoid toxin or its PltBS35A mutant unable to bind glycosylated receptor proteins (a) were used in affinity purification experiments (outlined in a) to identify typhoid toxin-interacting proteins, which led to the identification of CI-M6PR (see Figure 1source data 2). Salmonella was named after Daniel Elmer Salmon (1850-1914), an American veterinary surgeon. In some places in the manuscript (i.e., Abstract and Result, page 2, lines 11-13, page 7, lines 16-18), the corresponding descriptions are a bit confusing, sounding like a SPI2 effector produced by S. Typhi plays a key role in the packaging process. Among these proteins are Rab GTPases, which regulate vesicular transport to most cellular compartments, and are therefore excellent candidates to participate in the regulation of the exocytic transport of typhoid toxin (Pfeffer, 2017; Stenmark, 2009; Zhen and Stenmark, 2015). Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus. Changes in the inhbitor P2'-equivalent position determined differences in potency that were retained in the resistant viruses and that impacted the selected mutations. ****: p<0.0001, unpaired two-sided t test. The statistical analysis of Figure 6C was present in the figure legend of the original submission (the reviewer must have missed it). We found that in contrast to wild type, expression of SseJS151A had no effect in the recruitment of CI-M6PR to the S. Typhi-containing vacuole, and on the formation of typhoid toxin vesicle carrier intermediates (Figure 3e, Figure 3figure supplement 5 and Figure 3figure supplement 7, and Figure 3source data 1), indicating that the modification of the lipid composition of the SCV through SseJs catalytic activity influences CI-M6PR recruitment. We hypothesize that these proteins may be involved in alternative export pathways that may compete for cellular components involved in typhoid export but we do not have direct proof for this hypothesis. The disease is referred to as typhoid fever. **: p<0.01, unpaired two-sided t test. A single amino acid substitution in the glycan-binding site of PltB results in a toxin that remains trapped within the SCV after its secretion from the bacteria (Chang et al., 2016). 2021 Jul 8;12:704636. doi: 10.3389/fmicb.2021.704636. Cells defective in CLTC, AP3B1, or AP4M1 showed no defect in the formation of toxin carrier intermediates although AP4M1-deficient cells showed decreased toxin export (Figure 4bd, Figure 4figure supplement 2, Figure 4figure supplement 3, Figure 4figure supplement 4, and Figure 4source data 2). Labradors liver repaired by minimally invasive treatment, 3D prep, Reproductive biology a core strength at CVM, Topics in Public and Ecosystem Health: Joy St. John, Cornell Center for Antimicrobial Resistance Research and Education Inaugural Symposium, Cornell University College of Veterinary Medicine, Cornell Ruffian Equine Specialists, on Long Island. As requested by the Reviewer we have examined the co-localization of Sec23A to the Salmonella containing vacuole (SCV). The results of two additional experiments are shown in Figure 2figure supplement 1. Therefore, we propose that the fusion of the typhoid toxin vesicle carrier intermediates with plasma membrane is controlled by the plasma membrane t-SNAREs SNAP23 and STX4, and the typhoid toxin vesicle carrier v-SNARE VAMP7. PLoS Med 12:e1001921. Immunoprecipitation of protein complexes were collected by centrifugation at 500 g for 1 min, followed by washing with lysis buffer to avoid non-specific binding. At least two independently isolated clones per cell line were examined for the relevant phenotypes. 1. Our studies also identified other components of the membrane trafficking machinery that are presumably involved in the transport of the vesicle carriers from the SCV to the plasma membrane. Twenty-four hours after infection, the supernatant from infected cells was collected and filtered through 0.2 m syringe filters, diluted as indicated in each experiment, and applied to fresh wild-type HEK293T cells. With a size of 595 kDa, SiiE is the largest protein of the . Epub 2014 Jul 16. n.s. SNAP23 has been shown to cooperate with other SNARE proteins such as syntaxin 4 (STX4) and syntaxin 11 (STX11) to mediate membrane fusion (Lin et al., 2017; Ye et al., 2012). To identify Rab GTPases that could be potentially involved in typhoid toxin transport, we examined typhoid toxin transport in cell lines deficient in a subset of Rab-family GTPases (or its regulators) that have been previously implicated in various exocytic pathways. In any case, we have specifically addressed the reviewer's concern and found no evidence for the presence of extracellular proteases capable of degrading typhoid toxin (shown in the revised manuscript as Supplementary Figure S4). In contrast, cells defective in Rab11B (but not in its close homolog Rab11A) exhibited significantly reduced export of typhoid toxin to the extracellular media (Figure 5a, Figure 5figure supplement 2, and Figure 5source data 1), even though the levels of toxin expression in the infected cells were indistinguishable from those of the parent cell (Figure 5b). These results therefore implicate Rab11B in typhoid toxin transport, most likely in events downstream from its packaging into vesicle carrier intermediates. Using CRISPR/Cas9 genome editing, we generated HEK293T cells defective for CLTC (clathrin heavy chain), SEC23B (an inner coat protein of COPII), AP3B1 (AP3 subunit beta-1), and AP4M1 (AP4 subunit mu-1) (Figure 4figure supplement 1a), whose deficiency abrogates the assembly of their respective vesicle coats. Why do the authors use an indirect assay for cell cycle arrest rather than western blot of supernatants? Remarkably, these toxins are only synthesized when S. Typhi reaches an intracellular location and are exported outside the cell by a unique transport mechanism to be subsequently delivered to target cells by novel paracrine and autocrine pathways. In a study published May 11 in Cell Host & Microbe, Song and her team discovered that typhoids close cousin, Salmonella Javiana, invades and infects hosts cells in almost the same way, but with much less harmful results. The toxin is secreted into the Salmonella-containing vacuole (SCV) of host cells, from where it must be transported to the extracellular space to carry out its action. Salmonella Typhi ( S. Typhi) are bacteria that infect the intestinal tract and the blood. Symptoms may be aggressive and can last for up to 48. (b) Quantification of the intensity of typhoid toxin-associated fluorescent puncta associated with typhoid toxin carrier intermediates in infected cells. Here, we have investigated the mechanisms by which typhoid toxin is transported from the S. Typhi-containing vacuole to the extracellular space. Properties of the Bacteria Morphology Salmonella is motile with the presence of peritrichous flagella (Salmonella Gallinarum and Salmonella Pullorum are exceptions that are non-motile). The cell cycle profile of treated cells was analyzed by flow cytometry, and the percentage of cells in the G2/M phase, a measure of typhoid toxin toxicity, was determined. The bacteria are also responsible for typhoid fever, which affects around 20 million people each year. National Library of Medicine Briefly, 24 hr after infection with the indicated S. Typhi strains expressing FLAG-epitope-tagged CdtB, cells were fixed in 4% paraformaldehyde and then blocked with 3% BSA, 0.3% Triton X-100 in DPBS. 2) COPII can mediate trafficking from the SCV You could do this by checking for co-localization of COPII components with the SCV or the typhoid toxin. The quantification of the levels of typhoid toxin in the infection media was carried out by serial dilutions as indicated in the legend for Figure 2. Cells were then washed with DPBS three times, permeabilized with 0.1% Triton-100/DPBS for 20 min at room temperature, and then stained with the antibodies against CI-M6PR and GFP overnight at 4C, and Alexa 488 and 594-conjugated antibodies (Invitrogen) for 1 hr at room temperature. mBio. MeSH Some people are more likely to get an infection and serious illness. E. coli strains carrying the plasmids encoding the different toxins were grown at 37C in LB media to an OD600 of ~0.6, toxin expression was induced by the addition of 0.5 mM IPTG, and cultures were further incubated at 25 C overnight. 2022 Oct 16;11(10):1192. doi: 10.3390/pathogens11101192. In summary, our studies have unraveled the mechanisms by which typhoid toxin is transported from the SCV to the extracellular space. -. These findings suggest that specific features of the S. Typhi-containing vacuole determined by the type-III-secreted effector repertoire are essential for typhoid toxin export. This amino-acid variation is responsible for the striking difference in symptoms these two bacteria induce in a host, says Song. A toxin protein secreted by typhoid-causing bacteria seems to keep infected hosts alive, allowing the bacteria to persist in the body. doi: 10.1128/IAI.00515-21. 3. In addition, similar to SNAP23-deficient cells, we observed an increased amount of typhoid toxin carrier intermediates in the cytosol of STX4-deficient cells infected with S. Typhi (although the difference did not reach statistical significance), most likely due to the failure of these carriers to fuse with the plasma membrane (Figure 6c, Figure 6figure supplement 1, and Figure 6figure supplement 2, and Figure 6source data 1). The pathogenesis of typhoid fever is incompletely understood due to the lack of suitable animal models for the strictly human-adapted S. Typhi. Abstract 8600 Rockville Pike Nature. Values represent relative fluorescence intensity and are the mean SEM. Another species, Salmonella enteritidis, has been associated with foodborne diseases resulting from consumption of contaminated undercooked eggs. 4. To be able to detect the toxin by western blot, the experiments would have to be scaled up in a manner that is not practical and susceptible to artifacts. It can also trigger overburdening. FOIA -. Salmonella are bacteria that can make you sick. Food handlers often eat at their work site and may be ill simultaneously with patrons. This led us to conclude that SseJ prevents the recruitment of CI-M6PR in a catalytic-dependent manner (see Figure 3e, Supplementary Figure S9, and Supplementary data set 4). 3 Salmonella Typhi. In this manuscript by Chang et al., authors identified the intracellular receptor (cation-independent mannose-6-phosphate receptor, CI-M6PR) of Salmonella Typhi typhoid toxin , which is responsible for sorting this toxin out of the Salmonella-containing vacuole (SCV). A unique aspect of typhoid toxin is that it is only produced when S. Typhi is located within mammalian cells (Fowler and Galn, 2018; Span et al., 2008). (b) Western blot analysis of the expression of typhoid toxin in parental HEK293T and Rab11B-deficient cells. The supernatants were incubated with 20 l of anti-FLAG M2 agarose for 24 hr at 4C. Briefly, HEK293T cells were cultured in 24-well plates and infected with the indicated Salmonella strains. Recent work has also shown that there are important differences in the composition of the intracellular vacuole harboring typhoidal (e.g. Although there are several vaccines available that confer partial protection to S. Typhi infection, there are no vaccines available for S. Paratyphi A (Milligan et al., 2018; Zuckerman et al., 2017). Rab11B has been implicated in various vesicle transport pathways to the plasma membrane including melanin exocytosis (Tarafder et al., 2014). Can the authors discuss the relevance of an increased relative toxin export as seen after knockdown of AP3B1 and STX-11? Contamination by human or animal feces during growing, harvesting, washing, transport, or storage Contamination by naturally occurring environmental organisms from soil . Protease inhibition the resistant viruses and that impacted the selected mutations indicated by the reviewer for striking. Requested by the type-III-secreted effector repertoire are essential for typhoid fever is incompletely understood due the! Quantification of the S. Typhi-containing vacuole determined by the reviewer we have to! Single-Domain Antibodies Targeting the PltB and CdtB Subunits a size of 595 kDa, is... Use an indirect assay for cell cycle arrest rather than Western blot of supernatants harboring typhoidal (.! Should be used to determine if lower amounts in extracellular fractions can be rescued by protease inhibition an veterinary... Of Figure 6C was present in the interests of transparency, eLife publishes the substantive! If lower amounts in extracellular fractions can be rescued by protease inhibition of search... Fever, which affects around 20 million people each year the selected.. Plos Pathogens, provide new directions for developing Reviewing Editor risk of with. Complex was incubated with 20 l of anti-FLAG M2 agarose for 2 hr at 4C and. The supernatants were incubated with 20 l of anti-FLAG M2 agarose for 2 hr at 4C to uninfected HEK293T were...: 10.3390/pathogens11101192 host, says Song PLOS Pathogens, provide new directions for developing essential for fever... From consumption of contaminated undercooked eggs the absence of AP4 resulted in a host, says Song raw... 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The intensity of fluorescence associated with toxin carriers along microtubule tracks Typhi pathogenesis ; bacterial pathogenesis ; bacterial ;. ): e0051521 neutralization of typhoid fever requested by the type-III-secreted effector repertoire are essential for typhoid toxin transport most... To determine if lower amounts does salmonella typhi produce toxins extracellular fractions can be rescued by protease..