Perspectives on Oncolytic Salmonella in Cancer Immunotherapy-A Promising Strategy.

Since the first reported spontaneous regression of tumors in patients with streptococcus infection, cancer biological therapy was born and it evolved into today's immunotherapy over the last century. Although the original strategy was unable to impart maximal therapeutic benefit at the beginning, it laid the foundations for the development of immune checkpoint blockade and CAR-T which are currently used for cancer treatment in the clinics. However, clinical applications have shown that current cancer immunotherapy can cause a series of adverse reactions and are captious for patients with preexisting autoimmune disorders. Salmonellae was first reported to exert antitumor effect in 1935. Until now, numerous studies have proved its potency as an antitumor agent in the near future. In this review, we summarize the currently available data on the antitumor effects of Salmonella, and discussed a possibility of integrating Salmonella into cancer immunotherapy to overcome current obstacles.

Beneficial effect of immunobiotic strains on attenuation of Salmonella induced inflammatory response in human intestinal epithelial cells.

Probiotic bacteria have the ability to modulate host immune responses and have potent therapeutic functional effects against several diseases, including inflammatory diseases. However, beneficial effects of probiotics are strain specific and their interactions with host immune cells to modulate inflammatory response are largely unknown. Intestinal epithelial cells (IECs), which are the first line of defense against invading pathogens, and connects between commensals/probiotics and immune system; therefore, in this study, we used human IECs to assess the probiotic effects of three selected Lactobacillus strains in vitro. An HT-29 colonic epithelial cell and HT-29/blood mononuclear cells co-culture system were stimulated with Lactobacillus followed by Salmonella for different hours, after which the mRNA level of cytokines, ß-defensin-2 and negative regulators for TLR signaling and protein levels of ZO-1 and IκB-α were analyzed by real-time polymerase chain reaction and western blot analysis. L. brevis decreased Salmonella induced IL-6, IL-8, MCP-1 and IL-1ß levels, whereas L. pentosus suppressed IL-6 and MCP-1 in HT-29 cells. Moreover, L. brevis was able to increase the mRNA levels of A20, Tollip, SIGIRR and IRAKM, while L. pentosus reduced the levels of A20, and IRAKM in response to Salmonella. In addition, decrease in protein level of TNF-α and increase in mRNA level of IL-10 was observed in L. brevis and L. pentosus treated HT-29 cells. Lactobacillus strains were differentially modulated ZO-1 and p-IκB-α in HT-29 cells treated with Salmonella. Overall, the results of this study indicate that Lactobacillus strains attenuate Salmonella induced inflammatory responses through beneficial modulation of TLR negative regulators and the NF-κB pathway.

PART I: FIGHTING CANCER WITH DEADLY BACTERIA.

A 200-year-old observation might provide a new way to eliminate tumors by infecting cancer patients with bacteria. Kristie Nybo explores a new approach that could transform cancer treatment.

PART II: BREAKTHROUGHS IN TUMOR-SEEKING BACTERIA.

Newly engineered bacterial strains propel an immune-mediated cancer therapy toward clinical applications. Hannah Martin Lawrenz reports on the latest developments.

Vaccines for viral and bacterial pathogens causing acute gastroenteritis: Part II: Vaccines for Shigella, Salmonella, enterotoxigenic E. coli (ETEC) enterohemorragic E. coli (EHEC) and Campylobacter jejuni.

In Part II we discuss the following bacterial pathogens: Shigella, Salmonella (non-typhoidal), diarrheogenic E. coli (enterotoxigenic and enterohemorragic) and Campylobacter jejuni. In contrast to the enteric viruses and Vibrio cholerae discussed in Part I of this series, for the bacterial pathogens described here there is only one licensed vaccine, developed primarily for Vibrio cholerae and which provides moderate protection against enterotoxigenic E. coli (ETEC) (Dukoral(®)), as well as a few additional candidates in advanced stages of development for ETEC and one candidate for Shigella spp. Numerous vaccine candidates in earlier stages of development are discussed.

Reduction of subclinical Salmonella infection in fattening pigs after dietary supplementation with a ß-galactomannan oligosaccharide.

AIMS: To assess the efficacy of a ß-galactomannan oligosaccharide (ß-GMOS) for the control of Salmonella infection in fattening pigs. METHODS AND RESULTS: Three different doses (0.5, 3 and 2 kg ß-GMOS per ton of feed) were used during the entire period of growing in three similar and independent field trials carried out in a small fattening unit (≈ 100 pigs). Treatment was randomly assigned to half of the pens. Individual serum samples (20-25 per group) were collected at different times during the fattening period and a similar number of faecal samples during the fattening period and at slaughter. In addition, mesenteric lymph nodes were collected at slaughter. Herdcheck(®) Swine Salmonella ELISA was used for serological analyses, the ISO 6579:2002/Amd 1 : 2007 for bacteriology and the PFGE for molecular characterization of Salmonella strains. The addition of ≥ 2 kg t(-1) of ß-GMOS to the pig diet during the entire fattening period was associated with a reduction in Salmonella prevalence, shedding and seroconversion. CONCLUSIONS: Feed supplementation with ß-GMOS may be a useful complementary tool for the control of salmonellosis in fattening pigs. SIGNIFICANCE AND IMPACT OF THE STUDY: ß-GMOS may be a complementary way of reducing Salmonella shedding and infection in fattening pigs.

Passive immunity stimulated by vaccination of dry cows with a Salmonella bacterial extract.

BACKGROUND: Diarrhea because of Salmonella infection is a cause of neonatal calf diarrhea. The stimulation of passive immunity in the calf by vaccinating the dam for Salmonella has shown some success in previous studies; however, there are no data on the use of currently licensed vaccines in the United States. OBJECTIVE: To determine whether vaccinating cows at dry-off with a commercially available Salmonella bacterial extract would stimulate Salmonella-specific antibodies in the colostrum of cows at calving and whether these antibodies would be transferred to the calf. ANIMALS: Sixty Holstein cattle and 59 calves from a herd presumed to be naïve to Salmonella. METHODS: Prospective clinical trial. Thirty cows were vaccinated at dry-off with a Salmonella enterica serovar Newport bacterial extract and again 4 weeks later. An additional 30 cows received only saline. Calves fed fresh colostrum from their dam within 4 hours of birth had blood collected 24 hours later. RESULTS: Vaccinated cattle had increased Salmonella Newport antibody titers at calving in blood (P = .01) and colostrum (P = .011). Calves that received colostrum from vaccinated cattle also had significant increase in Salmonella antibodies (1.04 ± 0.03) as compared to calves born to unvaccinated cows (0.30 ± 0.02). CONCLUSIONS AND CLINICAL IMPORTANCE: The results indicate that the use of a commercially available Salmonella vaccine can stimulate antibodies that are passed on to the calf via colostral transfer. Further studies need to be done to determine whether these antibodies will offer protection against Salmonella challenge.

Salmonella gastroenteritis in children (clinical characteristics and antibiotic susceptibility): comparison of the years 1995-2001 and 2002-2008.

We document herein the prevalence and serotype distribution among Salmonella enterica strains isolated from children treated for diarrhea over two seven-year periods spanning 14 years. Four hundred and eight (1.38%) S. enterica cases were isolated among 29,601 diarrheal admissions. Among the Salmonella isolates, 63.7% were serogroup D and 29.9% were serogroup B. Overall, 21.7% of cases were under one year of age, with 2.1% being younger than three months. Bloody diarrhea was found in 18.8% of the cases. The resistance rates were 25.8%, 18.2%, 7.0%, 4.7%, and 0.3%, to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, ceftriaxone, and ciprofloxacin, respectively. In conclusion, our study has revealed that the predominance of Salmonella serogroup D continues. The clinical features of our patients were mostly mild, with no deaths or severe complications. While resistance to antimicrobial agents changes constantly, it is important to keep these strains under surveillance in order to formulate policies for the rational use of antimicrobial agents.

MyD88-dependent SHIP1 regulates proinflammatory signaling pathways in dendritic cells after monophosphoryl lipid A stimulation of TLR4.

We previously showed that monophosphoryl lipid A (MLA) activates TLR4 in dendritic cells (DCs) in a Toll/IL-1R domain-containing adaptor inducing IFN-ß (TRIF)-biased manner: MLA produced from Salmonella minnesota Re595 induced signaling events and expression of gene products that were primarily TRIF dependent, whereas MyD88-dependent signaling was impaired. Moreover, when tested in TRIF-intact/MyD88-deficient DCs, synthetic MLA of the Escherichia coli chemotype (sMLA) showed the same activity as its diphosphoryl, inflammatory counterpart (synthetic diphosphoryl lipid A), indicating that TRIF-mediated signaling is fully induced by sMLA. Unexpectedly, we found that the transcript level of one proinflammatory cytokine was increased in sMLA-treated cells by MyD88 deficiency to the higher level induced by synthetic diphosphoryl lipid A, which suggested MyD88 may paradoxically help restrain proinflammatory signaling by TRIF-biased sMLA. In this article, we demonstrate that sMLA induces MyD88 recruitment to TLR4 and activates the anti-inflammatory lipid phosphatase SHIP1 in an MyD88-dependent manner. At the same time, MyD88-dependent signaling activity at the level of IL-1R-associated kinase 1 is markedly reduced. Increased SHIP1 activity is associated with reductions in sMLA-induced IκB kinase α/ß and IFN regulatory factor 3 activation and with restrained expression of their downstream targets, endothelin-1 and IFN-ß, respectively. Results of this study identify a pattern that is desirable in the context of vaccine adjuvant design: TRIF-biased sMLA can stimulate partial MyD88 activity, with MyD88-dependent SHIP1 helping to reduce proinflammatory signaling in DCs.

Early eradication of persistent Salmonella infection primes antibody-mediated protective immunity to recurrent infection.

Typhoid fever is a systemic, persistent infection caused by host-specific strains of Salmonella. Although the use of antibiotics has reduced the complications associated with primary infection, recurrent infection remains an important cause of ongoing human morbidity and mortality. Herein, we investigated the impacts of antibiotic eradication of primary infection on protection against secondary recurrent infection. Using a murine model of persistent Salmonella infection, we demonstrate protection against recurrent infection is sustained despite early eradication of primary infection. In this model, protection is not mediated by CD4(+) or CD8(+) T cells because depletion of these cells either alone or in combination prior to rechallenge does not abrogate protection. Instead, infection followed by antibiotic-mediated clearance primes robust levels of Salmonella-specific antibody that can adoptively transfer protection to naïve mice. Thus, eradication of persistent Salmonella infection primes antibody-mediated protective immunity to recurrent infection.

Specific IgG activity against diarrheagenic bacteria in bovine immune milk and effect of pH on its antigen-binding activity upon heating.

Bovine colostrum and milk antibodies of calving and lactating cows immunized with a multivalent vaccine consisting of whole cells of three different species of pathogenic bacteria including four strains of enterotoxigenic Escherischia coli, five strains of enteropathogenic Esch. coli, three strains of enteroinvasive Esch. coli, two strains of Samonella typhi, and one strain each of Shigellia dysenteriae, Sh. sonnei and Sh. flexneri were generated, respectively. A significantly elevated activity and titre of specific IgG from bovine immune colostrum were seen for only 5 days after calving of immunized cows, however, the levels of specific IgG could be obtained continuously from the milk of immunized lactating cows until the 11th week of the entire experiment period. Subsequently, we observed that the high specific IgG activity in immune milk was relatively stable under pH 5.0-7.0 at 37 degrees C. Of importance, we identified that the specific IgG preserved its biological function for high antigen-binding activity at pH 5.5-6.5 for 30 min of heat treatment at 70 degrees C and for 350 s at 72 degrees C. Our findings suggest that the specific IgG from milk antibodies of immunized lactating cows may be used as an abundant source of hyper-immune products for prevention of multibacteria-induced diarrhea, however, the effect of pH on its antigen-binding activity upon heating should be carefully considered and designed.

Epidemiology of salmonella infections in pig units and antimicrobial susceptibility profiles of the strains of Salmonella species isolated.

One hundred and thirteen finishing pig units and 74 sow units in Catalonia, Spain, were examined to determine the prevalence of salmonella infections and the factors that could be associated with them. Pooled faecal samples were taken from the finishing units, and samples of faeces were collected from individual sows. The Salmonella isolates were serotyped, phage typed and examined for their antimicrobial susceptibility to 18 common antimicrobial drugs. In addition, blood samples from pigs on 141 farms were analysed by ELISA. In both the bacteriological and serological surveys, a questionnaire with 84 questions was completed for each farm. Salmonella species were isolated from 20 per cent of the finishing units and 24 per cent of the sow units; 14 serotypes were detected in the finishing pigs and 11 in the sows. More than 30 per cent of the strains were resistant to tetracycline, sulphonamides, ampicillin or streptomycin, and 69 per cent of the strains were resistant to three or more agents up to 10 compounds. Seventy-seven per cent of the farms had at least one seropositive animal, and 26 per cent of these farms had an individual seroprevalence of 50 per cent or more. The factors associated (P<0.05) with the excretion of Salmonella species in the finishing units were the practice of raising livestock other than pigs (odds ratio [OR]=6.18), the herd size (OR=5.87), and a past history of clinical salmonellosis (OR=4.97). For the sows, the factors associated (P<0.05) with the excretion of Salmonella species were having open-flushed drainage of sewage (OR=34.48), a lack of rodent control measures (OR=0.05) and the number of sows in the unit (OR=9.26). Factors associated with seropositivity in the finishing units were a lack of bird-proof nets (OR=0.30) and the use of water from private wells (OR=3.64).

Effects of carrageenans on the binding, phagocytotic, and killing abilities of macrophages to salmonella.

The effects of carrageenans (CGNs) on the host defense mechanisms of macrophages against Salmonella infection were examined in vitro by using macrophage-like J774.1 cells. Iota-CGN reduced the Salmonella-binding and phagocytotic activities of J774.1 cells, but it increased the killing activity of the cells. Kappa-CGN increased the binding activity, but reduced the killing ability. CGNs would affect the host defense mechanisms by modulating the macrophage functions.

Seroprevalence and antibiotic sensitivity of serotypes of Salmonella enterica in Greek pig herds.

Blood samples were taken from 50 pigs in each of 59 farrow-to-finish production herds and from 40 pigs in each of four of five registered multiplying herds. Samples of feed and faeces were also collected from 17 of the production herds and from the four multiplying herds. The sera were tested for antibodies to Salmonella enterica by the Danish mix-ELISA, and the organisms were isolated, serotyped and sensitivity tested by standard techniques. The average within-herd seroprevalence was 3.4 per cent and at least one pig tested seropositive in 21 of the 59 herds. In the multiplying herds, only a single seroreactor was detected. Salmonellae were isolated from only five of 95 feed samples, from two of the 17 herds sampled, Salmonella tennessee in four of five samples from one herd and an untypable strain in one of five samples from another. Four infected faecal samples were detected in four herds; they harboured Salmonella typhimurium, Salmonella bredeney or Salmonella london. No salmonellae were isolated from the samples of feed and faeces taken from the multiplying herds. The S london and S typhimurium had a low sensitivity to streptomycin, kanamycin and neomycin, and the S typhimurium also had low sensitivity to amoxycillin, ticarcillin, piperacillin, amoxycillin + clavulanic acid, cefalotin and cefoperazone. The other isolates were sensitive to all the antimicrobial agents tested.

Evaluation of an autogenous Salmonella bacterin and a modified live Salmonella serotype Choleraesuis vaccine on a commercial dairy farm.

OBJECTIVE: To compare the efficacy of a Salmonella bacterin and a modified live Salmonella ser. Choleraesuis vaccine on a commercial dairy. ANIMALS: 450 cows in late gestation and 80 calves. PROCEDURE: Group-1 cows (n = 150) were vaccinated once with a modified live S. Choleraesuis (serogroup C1) strain 54 (SC54) vaccine, group-2 cows (150) were vaccinated on enrollment and 30 days later with a Salmonella ser. Montevideo (serogroup C1) bacterin, and group-3 cows (150) served as unvaccinated controls. One gallon of colostrum harvested from the first 80 cows to calve was fed to each calf. Outcome assessments included fecal shedding of Salmonella spp for the first 10 days after parturition (cows) or birth (calves), milk production, involuntary culling rate, mastitis incidence, antimicrobial use, and mortality rate. RESULTS: Salmonellae were isolated from 306 of 309 (99%) cows and 64 of 74 (86.5%) calves. Shedding frequency was less in SC54-vaccinated cows and calves that received colostrum from those cows, compared with the other groups, and vaccination was specifically associated with less shedding of serogroup C1 salmonellae. Production data were similar among groups. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination of pregnant cows with an autogenous Salmonella bacterin had no effect on fecal shedding of salmonellae, whereas vaccination with a modified live S. Choleraesuis vaccine reduced the frequency of fecal shedding of serogroup C1 salmonellae during the peripartum period. A commercial S. Choleraesuis vaccine licensed for use in swine may be more efficacious than autogenous Salmonella bacterins on dairies infected with serogroup C1 salmonellae.

[The corrective action of antibodies in experimental intestinal dysbacteriosis]. / Korektiruiushchee deistvie antitel pri éksperimental'nom kishechnom disbakterioze.

As shown in this work, antisera obtained after the immunization of animals with vaccines, prepared from Salmonella minnesota strain R595 (Re mutant) or Escherichia coli O14 having enterobacterial common antigen (ECA), as well as human antisera with elevated titers of antibodies to Re glycolipid or to LPS O14 (ECA), inhibited the development of experimental intestinal dysbacteriosis in white mice, induced by the administration of ampyox in large doses. The degree of the inhibiting action of the antisera was proportional to antibody titers, which was indicative of the fact that antibodies possibly played some role in the regulation of the amount of intestinal microflora.

Dietary zinc-methionine enhances mononuclear-phagocytic function in young turkeys. Zinc-methionine, immunity, and Salmonella.

The ability of dietary zinc-methionine (Zn-Met) to enhance mononuclear-phagocytic function against Salmonella arizona and enteritidis was investigated in young turkeys. Feed/gain and body wt gain at 21 d of age were not affected by Zn-Met. The addition of 30 or 45 ppm Zn from Zn-Met to a Zn adequate diet significantly increased cutaneous basophil hypersensitivity to phytohemagglutinin-P. The clearance of intravenously administered S. enteritidis from blood was not affected by 30 ppm of supplemental Zn from Zn-Met. However, 30 ppm Zn from Zn-Met increased the reduction of intravenously administered S. arizona from spleen. Percentages of myeloid and mononuclear-phagocytic cells before and after S. enteritidis infection were not affected by supplemental Zn-Met. Turkeys supplemented with Zn-Met showed enhanced in vitro phagocytosis of S. enteritidis by Sephadex-elicited abdominal exudate cells. The phagocytosis of S. arizona was unaffected by Zn-Met.

[The use of monoclonal antibodies to lipid A for the correction of the hemodynamic disorders in endotoxemia]. / Ispol'zovanie monoklonal'nykh antitel k lipidu A dlia korrektsii gemodinamicheskikh narushenii pri éndotoksemii.

Endotoxemia induced by gram-negative bacteria leads to endotoxic shock pathogenetically stemming from the integral component of the bacterial wall--lipid A. The study made to define the ability of lipid A monoclonal antibodies to correct hemodynamic disturbances due to endotoxemia in dog experiments showed the efficacy of the antibodies administration. ReLPS isolated from Salmonella Minnesota was used as an antigen. Administration of the complex monoclonal antibodies-endotoxin caused no hemodynamic impairment.

Titres of class-specific antibodies against Shigella and Salmonella lipopolysaccharide antigens in colostrum and breast milk of Costa Rican, Swedish and Vietnamese mothers.

Enzyme immunoassays (EIA) were used to estimate titres of class-specific antibodies against purified and chemically defined phenol-water-extracted lipopolysaccharide (LPS) antigens of Salmonella serogroup B (BO), Shigella dysenteriae type I, Plesiomonas shigelloides (the same O-antigen as Shigella sonnei) and Shigella flexneri Y. Titres in colostrum and breast milk of Swedish, Vietnamese and Costa Rican mothers from various socioeconomic conditions were compared. The antibodies were mainly of the IgA isotype. IgM antibodies were also present, but only very low concentrations of IgG were found. In Costa Rican mothers, the IgA antibody titres were significantly higher (P less than 0.05) in women of low and middle socioeconomical conditions than were those in mothers of high socioeconomical level. The low titres in the last group were comparable to those found in Swedish mothers. The IgA antibody titres found in Vietnamese mothers were similar to those of Costa Rican mothers from the low and middle socioeconomic conditions, being highest against S. flexneri Y LPS. The IgM antibody titres were also highest in Vietnamese mothers, immediately followed by the Costa Rican mothers of low socioeconomic conditions. The low IgM titres in the Costa Rican women of high socioeconomic level were comparable to those seen in Swedish mothers. The results suggest that, in Costa Rica and Vietnam, S. flexneri is the most prevalent Shigella sp. causing infection and that Salmonella serogroup B infections are rare in all three countries. The results also show that the antibody repertoire in colostrum and breast milk varies. Furthermore, in addition to the prevalence of a specific micro-organism in a determined geographical area, such differences may be associated mainly with exposure to certain pathogens in particular socioeconomic conditions.