Combinations but Not a Single PlpE Epitope Induces Host Protective Immunity against Pasteurella multocida.

Pasteurella multocida is the causative agent of a wide range of diseases (pasteurellosis) and a zoonotic pathogen in humans. Recombinant subunit vaccines are hot spots in recent pasteurellosis vaccine development. A chimeric vaccine is also constructed for rabbit hemorrhagic disease virus (RHDV) protective antigen VP60 chimeric with fragments of Pasteurella multocida protective antigen PlpE. The protective efficacy of the chimeric vaccine against P. multocida is not as high as that of PlpE, and the reason is not well known. In this study, we analyzed the linear B-cell epitopes of PlpE and then assessed the protective efficacy of these epitopes and their combinations. It was found that the immunodominant region of PlpE was mainly located in the region between the 21st to the 185th amino acids from the N terminus. Overlapping peptide scanning results demonstrated that this region contained six nonoverlapping epitopes, and epitope E was the predominant epitope. Chimeric protein antigens were constructed of single nonoverlapping PlpE epitopes or their combinations chimeric with the RHDV VP60 P domain. Immunization with recombinant antigen chimeric with a single PlpE epitope exhibited poor immunoprotection, whereas immunization with recombinant antigen chimeric with PlpE epitope combinations (epitopes A and E; epitopes C and E; epitopes A, C, and E; and epitopes B, D, and F) exhibited significant immunoprotection. In a word, P. multocida protective antigen PlpE contained six nonoverlapping linear B-cell epitopes, and combinations but not a single epitope induced host protective immunity. Our work will give help for future chimeric vaccine design.

MAIT cells in bacterial infectious diseases: heroes, villains, or both?

Due to the aggravation of bacterial drug resistance and the lag in the development of new antibiotics, it is crucial to develop novel therapeutic regimens for bacterial infectious diseases. Currently, immunotherapy is a promising regimen for the treatment of infectious diseases. Mucosal-associated invariant T (MAIT) cells, a subpopulation of innate-like T cells, are abundant in humans and can mount a rapid immune response to pathogens, thus becoming a potential target of immunotherapy for infectious diseases. At the site of infection, activated MAIT cells perform complex biological functions by secreting a variety of cytokines and cytotoxic substances. Many studies have shown that MAIT cells have immunoprotective effects because they can bridge innate and adaptive immune responses, leading to bacterial clearance, tissue repair, and homeostasis maintenance. MAIT cells also participate in cytokine storm generation, tissue fibrosis, and cancer progression, indicating that they play a role in immunopathology. In this article, we review recent studies of MAIT cells, discuss their dual roles in bacterial infectious diseases and provide some promising MAIT cell-targeting strategies for the treatment of bacterial infectious diseases.

A novel live attenuated duck Tembusu virus vaccine targeting N7 methyltransferase protects ducklings against pathogenic strains.

Duck Tembusu virus (DTMUV), an emerging pathogenic flavivirus, causes markedly decreased egg production in laying duck and neurological dysfunction and death in ducklings. Vaccination is currently the most effective means for prevention and control of DTMUV. In previous study, we have found that methyltransferase (MTase) defective DTMUV is attenuated and induces a higher innate immunity. However, it is not clear whether MTase-deficient DTMUV can be used as a live attenuated vaccine (LAV). In this study, we investigated the immunogenicity and immunoprotection of N7-MTase defective recombinant DTMUV K61A, K182A and E218A in ducklings. These three mutants were highly attenuated in both virulence and proliferation in ducklings but still immunogenic. Furthermore, a single-dose immunization with K61A, K182A or E218A could induce robust T cell responses and humoral immune responses, which could protect ducks from the challenge of a lethal-dose of DTMUV-CQW1. Together, this study provides an ideal strategy to design LAVs for DTMUV by targeting N7-MTase without changing the antigen composition. This attenuated strategy targeting N7-MTase may apply to other flaviviruses.

Vitamin D3 can effectively and rapidly clear largemouth bass ranavirus by immunoregulation.

Largemouth bass ranavirus (LMBV) is a highly destructive pathogen that causes significant mortality rates among largemouth bass populations. Unfortunately, there is a dearth of drug development efforts specifically aimed at treating LMBV. To address this, our study sought to investigate the potential effectiveness of incorporating varying doses of VD3 into the diet as a treatment for LMBV. Through qRT-PCR and semi-qPCR, we observed significant suppression and clearance of LMBV pathogens in largemouth bass fed with 15000 IU/Kg and 20000 IU/Kg of VD3 within 14 days. In addition, VD3 treatment significantly increased the expression levels of key immune-related genes such as IL-1ß, IFN-γ, Mx, and IgM. Encouragingly, we observed that VD3 significantly increased antioxidant and immune activities such as TSOD, TAOC and C3 in serum and maintained total protein levels. Additionally, tissue pathology sections highlighted a dose-dependent relationship between VD3 supplementation and tissue damage, with the 15000 IU and 20000 IU groups exhibiting minimal damage. In conclusion, a reasonable concentration of VD3 effectively reduced LMBV replication and tissue damages, while improved immune-related genes expression and serum biochemical indices. These findings declare the considerable therapeutic potential of VD3 supplementation for combating LMBV disease and provide an alternative treatment option for fish farming.

Novel Hepatic Schistosomula Antigens as Promising Targets for Immunodiagnosis and Immunoprotection of Schistosomiasis japonica.

BACKGROUND: Antigens of migrating schistosomula are promising candidates as schistosomiasis vaccine targets, since immune attack on hepatic schistosomula would interrupt the parasites life cycle and reduce egg burden on the host. METHODS: In this study, we report a collection of Schistosoma japonicum schistosomula proteins (SjScPs) that are highly expressed in hepatic schistosomula. The expression characteristics, antigenicity and immune protection of these proteins were studied by western blot, ELISA, immunofluorescence and challenge assays. RESULTS: We found that several of these SjScPs were highly antigenic and could effectively stimulate humoral immune responses in both human and other mammalian hosts. In particular, SjScP25, SjScP37, SjScP41, SjScP80, and SjScP88 showed high potential as biomarkers for schistosomiasis immunodiagnosis. Furthermore, we demonstrated that immunization with several of the recombinant SjScPs were able to protect mice from S japonicum challenge infection, with SjScP25 generating the most protective results. CONCLUSIONS: Our work represents a group of novel schistosome immunogens, which may be promising schistosomiasis japonica diagnosis and vaccine candidates.

Cell delivery systems: Toward the next generation of cell therapies for type 1 diabetes.

Immunoprotection and oxygen supply are vital in implementing a cell therapy for type 1 diabetes (T1D). Without these features, the transplanted islet cell clusters will be rejected by the host immune system, and necrosis will occur due to hypoxia. The use of anti-rejection drugs can help protect the transplanted cells from the immune system; yet, they also may have severe side effects. Cell delivery systems (CDS) have been developed for islet transplantation to avoid using immunosuppressants. CDS provide physical barriers to reduce the immune response and chemical coatings to reduce host fibrotic reaction. In some CDS, there is architecture to support vascularization, which enhances oxygen exchange. In this review, we discuss the current clinical and preclinical studies using CDS without immunosuppression as a cell therapy for T1D. We find that though CDS have been demonstrated for their ability to support immunoisolation of the grafted cells, their functionality has not been fully optimized. Current advanced methods in clinical trials demonstrate the systems are partly functional, physically complicated to implement or inefficient. However, modifications are being made to overcome these issues.

Treatment of protoscoleces with gamma radiation: Potential immunoprotective effect against experimental murine echinococcosis.

Cystic echinococcosis (CE) is one of the most important zoonotic diseases with a worldwide distribution. It is caused by the larval stage of the dog tapeworm "Echinococcus granulosussensu lato" and constitutes a major economic and public health problem in several countries. Protoscoleces are one component of this larval stage that can interact with both definitive and intermediate hosts. The aim of the present study was to investigate the potential role of using a radio-attenuated form of these protoscoleces for immunoprophylaxis against experimental murine echinococcosis. However, mice were immunized twice at 15-day intervals with gamma (γ) irradiated protoscoleces at doses of 0.4, 0.8, 1.2 and 1.4 kGy then challenged with the intact parasites. Macroscopic and histological analyses with cytokine measurements were performed in order to estimate the number and diameter of cysts, microscopic changes and cytokine profile. An improvement in protection against the challenge dose was observed with increasing dose, giving percentages of 47.7, 49, 55.23 and 70.6%, for the 0.4, 0.8, 1.2 and 1.4 kGy-groups respectively. These data suggest that immunization with radio-attenuated protoscoleces may induce satisfactory protective immunity by reducing successfully the formation of cysts, caused by challenge infection.

Immune enhancement effects of inactivated vaccine against extracellular products of Aeromonas caviae AC-CY on crucian carp.

Aeromonas caviae is a zoonotic pathogen that can cause disease in aquatic organisms and mammals, including humans, and it is widespread in nature, especially in freshwater environments. Previous research has reported that extracellular products (ECPs) secreted by pathogens during growth are effective protective antigens that can induce the host immune response and protect the host from pathogens. However, little is known about how ECPs enhance immunity. Here, we prepared extracellular products by the cellophane plate method, determined the total protein concentration, and analysed the protein composition of the extracellular products by SDS-PAGE. Subsequently, their enzyme activity and pathogenicity were evaluated separately. Crucian carp were randomly divided into four groups to receive formalin-inactivated A. caviae vaccine (FKC), ECPs mixed with the same amount of Freund's complete adjuvant, the same amount of ECPs mixed with an equal volume of A. caviae inactivated vaccine (FKC + ECPs), sterile PBS alone via intraperitoneal injection. On Days 7, 14, 21, and 28 after immunization, the expression levels of IgM, SOD, and CAT and the lysozyme (LYS) activity in the serum were detected by ELISA, and the relative expression levels of the TNF-α, IFN-γ, IL-1ß, and IL-10 genes in the liver, kidney, spleen, intestine, and gills were measured by qPCR. The extracellular products generated five clearly visible protein bands and exhibited lipase, protease, amylase, DNase and lysozyme but no urease or lecithinase activities. In addition, the median lethal doses of A. caviae and ECPs to crucian carp were 411.64 µg/fish and 1.6 × 105 CFU/mL, respectively. Compared with those of the control group, the IgM, SOD, and CAT contents and serum LYS activity were significantly increased in the experimental groups, and the qRT-PCR results showed that the relative expression levels of TNF-α, IFN-γ, IL-1ß, and IL-10 genes in the liver, kidney, spleen, and intestine were significantly increased after injection immunization. In addition, the relative immune protection rates of the three experimental groups were 60%, 65%, and 45%, all of which were significantly higher than those of the control group. Collectively, our findings show that the extracellular products of A. caviae can be used as a vaccine to significantly improve the immune level of crucian carp and have obvious anti-infection ability. This may represent a promising approach to prevent and control infection by A. caviae and provides strong theoretical support for the development of new inactivated vaccines.

Screening the potential part of the G protein antigen is an achievable strategy to improve the immune effect of DNA vaccine against MSRV infection.

DNA vaccines, as an effective prophylactic technology to induce both humoral and cellular immune responses, have already been widely studied to prevent and control viral and bacterial infections in aquaculture. To find a more effective and safer way to control Micropterus salmoides rhabdovirus (MSRV) infection in largemouth bass, two different DNA vaccines expressing partial (pcDNA3.1-G2) and full-length (pcDNA3.1-G) of the MSRV G protein were developed and injected intramuscularly with different doses. The immune effect was comprehensively compared and evaluated by detecting immune-related parameters including serum antibody levels, immune-related physiological indexes, immune-related gene expression and relative survival rates in this study. The results showed that compared with the pcDNA3.1-G vaccine, the pcDNA3.1-G2 vaccine induced higher serum antibody levels, a lower nonspecific immune response in serum (ACP, SOD and T-AOC activities), higher immune-related gene expression and a higher relative survival rate. Moreover, the immune effect of pcDNA3.1-G2-vaccinated fish showed gradually higher with the increasing pcDNA3.1-G2 concentration, especially in pcDNA3.1-G2 (10µg/per fish) group, the relative survival rate reached to 82.5%, which was significant higher (p < 0.05) than pcDNA3.1-G (10µg/per fish) group. This study indicated that screening the potential core part of an antigen is an achievable strategy to improve the immunogenicity and immunoprotective effect of DNA vaccine.

Immunoprotection against mixed Eimeria spp. infections in goat kids induced by X-irradiated oocysts.

Strategies to control goat coccidiosis traditionally rely on the use of management practices combined with anticoccidial treatments, and limited effort has been made, so far, to address immunological control of caprine Eimeria infections. Previously, we showed that monospecific immunization with X-Rad-attenuated Eimeria ninakohlyakimovae oocysts induced considerable immunoprotection upon challenge. In the present study, we conducted a similar vaccination trial but using a mixture of caprine Eimeria species typically present in natural infected goats. For immunization, sporulated oocysts were attenuated by X irradiation (20 kilorad). All infections were performed orally applying 105 sporulated oocysts of mixed Eimeria spp. per animal. In total, 18 goat kids were grouped as follows: (G1) immunized + challenge infected; (G2) primary + challenge infected; (G3) challenge infection control; and (G4) non-immunized/non-infected control. Overall, goat kids infected with attenuated oocysts (= immunized) shed less oocysts in the faeces and showed a lower degree of clinical coccidiosis than animals infected with non-attenuated oocysts. Animals of both challenge groups (G1 and G2) showed partial immunoprotection upon reinfection when compared to challenge infection control (G3). However, the degree of immunoprotection was less pronounced than recently reported for monospecific vaccination against Eimeria ninakohlyakimovae, most probably due to the complexity of the pathogenesis and related immune responses against mixed Eimeria spp. infections. Nevertheless, the data of the present study demonstrate that immunization with attenuated Eimeria spp. oocysts may be worth pursuing as a strategy to control goat coccidiosis.

Selenium: An Antioxidant with a Critical Role in Anti-Aging.

Aging is characterized by an imbalance between damage inflicted by reactive oxygen species (ROS) and the antioxidative defenses of the organism. As a significant nutritional factor, the trace element selenium (Se) may remodel gradual and spontaneous physiological changes caused by oxidative stress, potentially leading to disease prevention and healthy aging. Se is involved in improving antioxidant defense, immune functions, and metabolic homeostasis. An inadequate Se status may reduce human life expectancy by accelerating the aging process or increasing vulnerability to various disorders, including immunity dysfunction, and cancer risk. This review highlights the available studies on the effective role of Se in aging mechanisms and shows the potential clinical implications related to its consumption. The main sources of organic Se and the advantages of its nanoformulations were also discussed.

Development and evaluation of polymeric nanoparticles as a delivery system for snake envenoming prevention.

Polymer-based nanoparticles have become an appealing carrier for improving vaccine delivery efficiency. In this study, we investigated an interesting approach based on PLGA nanoparticles encapsulating Cerastes cerastes venom as an intranasal vaccine delivery system for snake envenomation prevention. Particles were synthesized by double emulsion solvent evaporation method and characterized for their size, morphology, distribution, and venom-nanoparticles interactions. An immunization trial was performed in mice by the intranasal route to evaluate the immune response, the reactogenicity, and the protective effect of this nanovaccine. The physicochemical and structural characteristics of Cc-loaded PLGA NPs revealed that the particles exhibited a spherical shape with a diameter of 370 nm, and a negatively charged surface with a zeta potential value of 19,9 mV. The immunization with Cc-PLGA NPs can induce a systemic innate and humoral immune response and confers protection against Cerastes cerastes venom (Cc) over than 6 LD50 with a cross-protection against Vipera lebetina venom (Vl) over than 5 LD50. Nano-encapsulation of Cc venom reduced its toxicity and the induced tissue alterations. Our results confirm that the nano-formulation Cc-PLGA NPs is a potent adjuvant system that improve the humoral immune response and provide protection against high lethal doses of viper venoms.

Mechanism by which the combination of SjCL3 and SjGAPDH protects against Schistosoma japonicum infection.

A vaccine is an important method to control schistosomiasis. Molecules related to lung-stage schistosomulum are considered potential vaccine candidates. We previously showed that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and cathepsin L3 (CL3) displayed differential expression in the lung-stage schistosomula of Schistosoma japonicum cocultured with host cells. In the present study, we prepared the two proteins and detected the protective effects of SjGAPDH by immunizing mice with this protein alone and in combination with SjCL3 with or without Freund's adjuvant. Then, we investigated the possible mechanisms underlying S. japonicum infection. The results showed that vaccination of adjuvanted SjGAPDH decreased the worm burden (37.8%) and egg load (38.1%), and the combination of adjuvanted SjGAPDH and SjCL3 further decreased the worm burden (65.6%) and egg load (70.9%) during Schistosoma japonicum infection. However, the immunization of a combination of adjuvant-free SjGAPDH and SjCL3 displayed a lower protective effect (< 15%) than those of the adjuvanted SjCL3, the adjuvanted SjGAPDH, and a combination of adjuvanted SjGAPDH and SjCL3. Flow cytometric results showed that the frequency of regulatory T cells (Tregs) was lower (P < 0.05) in the group with adjuvanted SjGAPDH and SjCL3 (2.61%) than the remaining groups. The enzyme-linked immunosorbent assay (ELISA) results indicated that except for the uninfected and infected control groups, the remaining groups displayed a Th1-type shift in immune responses. These results showed the immunization of SjGAPDH resulted in partial protection (approximately 38%); inoculation with a combination of SjCL3 and SjGAPDH in Freund's adjuvant resulted in a high immunoprotective effect (> 65%) against Schistosoma japonicum infection in mice, which was possibly caused by the reduced percentage of Tregs and a Th1-type shift in immune responses; and SjCL3 has no adjuvant-like effect, dissimilar to SmCL3.

Immunoprotective potential of Ayurvedic herb Kalmegh (Andrographis paniculata) against respiratory viral infections - LC-MS/MS and network pharmacology analysis.

INTRODUCTION: Immunity boosting has emerged as a global strategy to fight the SARS-CoV-2 pandemic situation. In India, AYUSH systems of medicine have been promoted as an immune-protection strategy. Andrographis paniculata (Burm. F) Nees (AP) mentioned in Ayurveda has been widely used for treating sore throat, flu, and upper respiratory tract infections which may provide possible novel therapeutic approaches, exclusively targeting SARS-CoV-2 and its pathways. OBJECTIVES: The present work uses liquid chromatography-tandem mass spectrometry (LC-MS/MS) metabolomics and combination synergy analysis based on network pharmacology to mine multimode evidence to understand the possible mechanism of action, diseases association, protein-protein interaction and major pathways involved therein. MATERIAL AND METHODS: Metabolite profiling was performed by Agilent QTOF LC-MS/MS system. Network pharmacology analysis was performed by using functional annotation analysis based on databases like Binding DB, STRING, DAVID and KEGG for further data mining. Further combination synergy was evaluated using "neighbourhood approach" and networks were constructed through Cytoscape 3.2.1. RESULTS: The molecules from kalmegh provides immune-protection and anti-viral response via involving different pathways, like toll-like receptor pathway, PI3/AKT pathway and MAP kinase pathways against COVID-19 infection. The KEGG analysis showed that in a vast majority of the most enriched pathways, AP were associated with viral infections and upper respiratory tract infections. CONCLUSIONS: The results suggest a synergy between andrographolide and other molecules identified as safe and efficacious anti-inflammatory agent having effects on upper respiratory tract infections and can significantly decrease the production of cytokines and pro-inflammatory factors in viral infections.

Recombinant Vibrio parahaemolyticus ghosts protect zebrafish against infection by Vibrio species.

Aquatic animals are frequently threated by bacterial pathogens. The most economic and efficient protection against bacterial infection are through vaccine immunization. The various serotypes of the pathogens, such as Vibrios, hurdle the development of the vaccines, especially polyvalent vaccines. Here, we demonstrate that recombinant bacterial ghost is a good candidate for multivalent vaccine. By expressing PhiX174 gene E alone or co-expressing the gene E with two genes encoding outer membrane proteins (VP1667 and VP2369) in V. parahaemolyticus, we generated the recombinant V. parahaemolyticus ghosts VPG and rVPGs respectively. Fish immunized with either VPG or rVPG showed increased survival against the infection by either V. parahaemolyticus or V. alginolyticus, with a better protective effect by immunization with rVPG. Our furthermore studies show that rVPG stimulates stronger innate immune responses by increasing the expression of tnfα, il1ß, il6, il8 and il10 as well as that of c3b, lyz, and tlr5, the key players linking the innate and adaptive immune responses upon microbial stimulation. In summary, VPG and rVPG can protect zebrafish against the infection from at least two Vibrio species, suggesting its potential value for further aquaculture vaccines development.

Dietary supplementation of marine yeast Yarrowia lipolytica modulates immune response in Litopenaeus vannamei.

The immunostimulatory potential of the marine yeast Yarrowia lipolytica (D1 and N6 strains) administered orally was evaluated in the white shrimp Litopenaeus vannamei. Yeasts and commercial glucans were mixed with a commercial feed to formulate diets with a 1.1% concentration of immunostimulants. The shrimp were fed daily for a period of 21 days. Weekly determinations were performed for immunological parameters in hemolymph, such as total hemocyte count (THC), lysozyme activity (LYZ), prophenoloxidase activity, antioxidant enzymatic activities (superoxide dismutase [SOD], catalase [CAT], and peroxidases), and bactericidal activity against Vibrio parahaemolyticus. Expression profiles of penaeidin (PEN), lysozyme (LYZ), and prophenoloxidase (proPO) immune genes were evaluated in hemocytes. In general, an increase in the immune parameters was observed in shrimp fed yeast diet compared to glucan and the control diets. Yarrowia lipolytica, especially strain N6, provided maximum immunostimulatory effects evidenced by the increase of immune parameters (THC, LYZ, SOD, CAT) and gene expression profile. In conclusion, this study demonstrated that Y. lipolytica had immunostimulatory effects and increased bactericidal activity in L. vannamei hemocytes against V. parahaemolyticus. These findings open the path for the potential application of Y. lipolytica-based immunostimulant for shrimp aquaculture.

An effective live attenuated vaccine against Streptococcus agalactiae infection in farmed Nile tilapia (Oreochromis niloticus).

Streptococcus agalactiae is an important pathogen associated with various aquatic animals, especially tilapia. Streptococcosis has greatly limited the healthy development of tilapia aquaculture in recent times. The development of novel effective vaccines is important for the prevention and control of streptococcosis in fish. We previously constructed a non-encapsulated S. agalactiae strain △cps by the in-frame deletion method. Here, we evaluated whether this mutant △cps is safe for tilapia and suitable for protection against streptococcosis. We observed that the △cps strain was non-pathogenic to tilapia, and there was no reversion of virulence when it was passaged in tilapia. Moreover, the △cps strain survived for at least 11 d in the main immune organs of tilapia. The tilapia vaccinated via intraperitoneal (IP) injection with △cps strain induced a high antibody titer, and the IgM antibody levels were significantly higher in the vaccinated group than in the control group. The vaccination protected tilapia against the S. agalactiae challenge with a relative percent survival of 90.47%. In addition, tilapia immunized with the △cps strain showed significantly higher expression level of IFN-γ, IL-1ß, MyD88, IgM, and MHC-Iα in the head kidney than those in the control during the entire observation period. The expression of MHC-IIß was inhibited during 1-7 d of immunization. These results revealed that the △cps strain is able to induce humoral and cell-mediated immune response in tilapia. Therefore, the strain △cps has a broad application prospect as a target for attenuation in vaccine development.

Sequential Versus Combined Heart-Liver Transplantation in the USA.

INTRODUCTION: Combined heart-liver transplantation (CHLT) has resulted in acceptable survival rates compared to orthotopic liver transplantation (OLT) alone and orthotopic heart transplantation alone. Using the US transplant registry, we compared outcomes following sequential and combined HLT. METHODS: We conducted a retrospective cohort study. De-identified data were obtained from the United Network Organ Sharing Registry. The primary outcome was patient survival from the date of OLT. Secondary outcomes included liver allograft survival and heart allograft survival. RESULTS: The study cohort included 301 CHLT recipients and six sequential heart-liver transplantation (SHLT) recipients. Patient survival after CHLT was 88% at 1 year, 84% at 3 years, and 82% at 5 years compared to 83%, 67%, and 50% in the SHLT group (p = 0.010). Liver allograft survival at 1, 3, and 5 years was 88%,83% and 82%, respectively, in the CHLT group compared to 83% and 67%, and 50%, respectively, in the SHLT group (p = 0.009). After OLT, heart allograft survival at 1, 3, and 5 years was 86%, 79%, and 74% in the CHLT group, respectively, compared to 83%, 67%, and 50% in the SHLT group (p = 0.037). CONCLUSIONS: Despite the limited size of the SHLT cohort, we found that CHLT was superior to SHLT in survival rate and graft survival. The better outcomes noted in CHLT may relate to immunoprotection provided by liver transplantation from the same donor.

Lactobacillus reuteri protects mice against Salmonella typhimurium challenge by activating macrophages to produce nitric oxide.

Lactobacillus reuteri, a typical intestinal symbiotic bacterium, plays an important role in maintaining intestinal flora stability and host health. However, the effect of Lactobacillus reuteri on peritoneal macrophages has not been thoroughly studied. Our study indicated that Lactobacillus reuteri could activate macrophages and that macrophages treated with Lactobacillus reuteri have an enhanced ability to phagocytose and to kill intracellular Salmonella typhimurium. Lactobacillus reuteri may reduce the inflammatory response caused by Salmonella typhimurium by regulating NO, thus effectively protecting mice against Salmonella typhimurium invasion and dissemination to the liver and spleen. Taken together, these data demonstrated the protective effect of Lactobacillus reuteri on macrophages and mice challenged with Salmonella typhimurium through in vitro and in vivo experiments.

Evaluation of the immunogenicity of an omp A and staphylococcal target of RNAIII activating fusion protein displayed on the surface of Escherichia coli.

The purpose of this investigation was to construct a recombinant Escherichia coli strain displaying the Staphylococcus aureus target of RNAIII activating protein (TRAP) on its surface, and to investigate the strain for its immunogenicity. The lpp'ompA and lpp'ompA-TRAP genes were fused by the overlap polymerase chain reaction and then ligated into expression plasmid pQE30 producing pLO and pLO-TRAP. These two recombinant plasmids were transformed into E. coli XL1-Blue, resulting in XL1-Blue/pLO and XL1-Blue/pLO-TRAP, which were induced to express protein. The expressed TRAP protein was displayed on the surface of XL1-Blue as judged by whole cell ELISA, flow cytometric analysis, and laser scanning confocal microscopy using the lpp'ompA surface display system. ICR mice were intramuscularly immunized with recombinant strains XL1-Blue/pLO and XL1-Blue/pLO-TRAP as well as recombinant protein TRAP. Immunized mice were assessed for anti-TRAP antibody and lymphocytes for secreted IL-4 and IFN-γ by ELISPOT and secreted IL-17A by indirect ELISA. Immunized mice were challenged with S. aureus Newman and HLJ23-1 strains. The results showed both XL1-Blue/pLO-TRAP and TRAP protein immunized mice to produce better cellular and humoral immunity than XL1-Blue/pLO and PBS injected mice.