Immunization with recombinant protein Ag43::UpaH with alum and 1,25(OH)2D3 adjuvants significantly protects Balb/C mice against urinary tract infection caused by uropathogenic Escherichia coli.

The majority of urinary tract infections (UTIs) are caused by uropathogenic Escherichia coli (UPEC). Designing a vaccine will certainly reduce the occurrence of infection and antibiotic resistance of the isolates. Antigen 43 (Ag43) and autotransporter H (UpaH) have been associated with the virulence of UPEC. In the present study, the efficacy of different formulations of a hybrid protein composed of Ag43 and UpaH with and without alum and 1,25(OH)2D3 (Vitamin D3) adjuvants were evaluated in mice model. A significant increase in IgG and cellular responses was developed against Ag43::UpaH as compared to the control mice. The addition of alum or a mixture of alum and Vitamin D3 to the protein significantly enhanced the serum IgG responses and tended to remain in a steady state until 6 months. In addition, the mentioned formulations produced significant amounts of IgG1, IL-4, and IL-17 as compared to the fusion protein alone. In addition to the mentioned formulations, the combination of protein with Vitamin D3 also resulted in significantly higher serum IgA and IFN-γ levels as compared to the fusion protein alone. Mice immunized with fusion plus alum and formulation protein admixed with both alum and Vitamin D3 significantly reduced the bacterial load in the bladders and kidneys of mice as compared to the control. In this study, for the first time, the ability of a novel hybrid protein in combination with adjuvants alum and Vitamin D3 was evaluated against UPEC. Our results indicated that fusion Ag43::UpaH admixed with alum and Vitamin D3 could be a promising candidate against UTIs.

Helicobacter pylori treatment: New perspectives using current experience.

Infection with Helicobacter pylori plays an essential role in the development of duodenal and gastric ulcers as well as in the pathobiology of gastric adenocarcinoma. Thus, successful elimination of the bacterium can reduce the risk of development of these diseases. Currently, most guidelines recommend standard triple therapy (amoxicillin+clarithromycin+proton pump inhibitor), although its efficacy is rapidly falling. Notably, traditional first-line therapy fails in almost 32% of H. pylori-carrying cases, suggesting the importance of choosing the best formulation for first-line therapy. Hence, due to the decreasing effectiveness of first-line treatment, we should be prepared to confront increasing H. pylori therapeutic defeat. Owing to increasing reports of antibiotic resistance worldwide, newer approaches and directions are necessary for managing this problematic infection. Developing and providing better anti-H. pylori strategies (probiotics, antibiotic therapy and non-traditional medicine) without using current clinical experience in treating the infection is impossible. Furthermore, development and examination of new preventive vaccines may also be a new therapeutic direction. Taken together, with regard to current experience, clinicians are highly recommended to consider all alternatives to eradicate H. pylori until a universal vaccine becomes practically available. This article aims to give an overview regarding the current status of H. pylori treatment, accordingly designing an actual overview to gain optimal strategies against this infection.

Plant-based vaccines against respiratory diseases: current status and future prospects.

INTRODUCTION: Respiratory infections have an enormous, worldwide epidemiologic impact on humans and animals. Among the prophylactic measures, vaccination has the potential to neutralize this impact. New technologies for vaccine production and delivery are of importance in this field since they offer the potential to develop new immunization approaches overriding the current limitations that comprise high cost, safety issues, and limited efficacy. Areas covered: In the present review, the state of the art in developing plant-based vaccines against respiratory diseases is presented. The review was based on the analysis of current biomedical literature. Expert commentary: Preclinical and clinical evaluations of several vaccine candidates against influenza, tuberculosis, respiratory syncytial virus, pneumonia, anthrax and asthma are discussed and placed in perspective.

A partially purified outer membrane protein VirB9-1 for low-cost nanovaccines against Anaplasma marginale.

Anaplasma marginale is a devastating tick-borne pathogen causing anaplasmosis in cattle and results in significant economic loss to the cattle industry worldwide. Currently, there is no widely accepted vaccine against A. marginale. New generation subunit vaccines against A. marginale, which are much safer, more efficient and cost-effective, are in great need. The A. marginale outer membrane protein VirB9-1 is a promising antigen for vaccination. We previously have shown that soluble recombinant VirB9-1 protein can be expressed and purified from Escherichia coli and induce a high level of humoral and cellular immunity in mice. In this study, we re-formulated the nanovaccines using the partially-purified VirB9-1 protein as the antigen and hollow nano-size silica vesicles (SV-100) as the adjuvant. We simplified the purification method to obtain the partially-purified antigen VirB9-1 with a six-fold higher yield. The new formulations using the partially-purified VirB9-1 protein achieved higher antibody and cell-mediated immune responses compared to the purified ones. This finding suggests that the partially-purified VirB9-1 protein performs better than the purified ones in the vaccination against A. marginale, and a certain level of contaminants in the protein antigen can be self-adjuvant and boost immunogenicity together with the nanoparticle adjuvant. This may lead to finding a "Goldilocks" level of contaminants. The new nanovaccine formulation using partially-purified antigens along with nanoparticle adjuvants offers an alternative strategy for making cheaper veterinary vaccines.

Strategies to develop vaccines of pediatric interest.

INTRODUCTION: The success of the vaccines available on the market has significantly increased interest in vaccine development. Areas covered: The main aim of this paper is to discuss the most important vaccines of pediatric interest that are currently being developed. New pneumococcal vaccines and vaccines against group B Streptococcus, Staphylococcus aureus and respiratory syncytial virus are analyzed in detail. Expert commentary: Advances in understanding human immunology, including human monoclonal antibody identification, sequencing technology, and the ability to solve atomic level structures of vaccine targets have provided tools to guide the rational design of future vaccines. It is likely that some of these vaccines will reach the market in the future and will thus partially contribute to the prevention of very severe diseases that significantly affect the morbidity and mortality of children. However, further studies in animals and several clinical trials in children must be performed before new vaccines become licensed.

Summary and Recommendations from the National Institute of Allergy and Infectious Diseases (NIAID) Workshop "Gonorrhea Vaccines: the Way Forward".

UNLABELLED: There is an urgent need for the development of an antigonococcal vaccine due to the increasing drug resistance found in this pathogen. The U.S. Centers for Disease Control (CDC) have identified multidrug-resistant gonococci (GC) as among 3 "urgent" hazard-level threats to the U.S. POPULATION: In light of this, on 29 to 30 June 2015, the National Institute for Allergy and Infectious Diseases (NIAID) sponsored a workshop entitled "Gonorrhea Vaccines: the Way Forward." The goal of the workshop was to gather leaders in the field to discuss several key questions on the current status of gonorrhea vaccine research and the path forward to a licensed gonorrhea vaccine. Representatives from academia, industry, U.S. Government agencies, and a state health department were in attendance. This review summarizes each of the 4 scientific sessions and a series of 4 breakout sessions that occurred during the one and a half days of the workshop. Topics raised as high priority for future development included (i) reinvigoration of basic research to understand gonococcal infection and immunity to allow intervention in processes essential for infection; (ii) clinical infection studies to establish parallels and distinctions between in vitro and animal infection models versus natural human genital and pharyngeal infection and to inform in silico modeling of vaccine impact; and (iii) development of an integrated pipeline for preclinical and early clinical evaluation and direct comparisons of potential vaccine antigens and adjuvants and routes of delivery.

Novel nanoparticle vaccines for Listeriosis.

In recent years, nanomedicine has transformed many areas of traditional medicine, and enabled fresh insights into the prevention of previously difficult to treat diseases. An example of the transformative power of nanomedicine is a recent nano-vaccine against listeriosis, a serious bacterial infection affecting not only pregnant women and their neonates, but also immune-compromised patients with neoplastic or chronic autoimmune diseases. There is a major unmet need for an effective and safe vaccine against listeriosis, with the challenge that an effective vaccine needs to generate protective T cell immunity, a hitherto difficult to achieve objective. Now utilizing a gold nanoparticle antigen delivery approach together with a novel polysaccharide nanoparticulate adjuvant, an effective T-cell vaccine has been developed that provides robust protection in animal models of listeriosis, raising the hope that one day this nanovaccine technology may protect immune-compromised humans against this serious opportunistic infection.

Current advances and challenges in the development of Acinetobacter vaccines.

Acinetobacter baumannii is a major cause of healthcare-associated infections worldwide with high morbidity and mortality. The clinical treatment of A. baumannii infections has become increasingly difficult because of the rapid emerging of multidrug and extremely drug resistant strains. Thus, there is an urgent need for the development of novel intervention strategies to combat this multidrug-resistant pathogen. Vaccine is one of the most effective medical measures for infection control and is likely to overcome the development of multidrug resistance by A. baumannii. Here we discussed the recent advances and potential challenges in development of A. baumannii vaccines with a focus on the 3 most important steps in the preclinical vaccine development: antigen selection, immune correlates of protection, and animal models for efficacy evaluation.

Evaluation of a vaccine formulation against Streptococcus pneumoniae based on choline-binding proteins.

Streptococcus pneumoniae has proteins that are attached to its surface by binding to phosphorylcholine of teichoic and lipoteichoic acids. These proteins are known as choline-binding proteins (CBPs). CBPs are an interesting alternative for the development of a cost-effective vaccine, and PspA (pneumococcal surface protein A) is believed to be the most important protective component among the different CBPs. We sought to use CBPs eluted from pneumococci as an experimental vaccine. Since PspA shows variability between isolates, we constructed strains producing different PspAs. We used the nonencapsulated Rx1 strain, which produces PspA from clade 2 (PspA2), to generate a pspA-knockout strain (Rx1 ΔpspA) and strains expressing PspA from clade 1 (Rx1 pspA1) and clade 4 (Rx1 pspA4). We grew Rx1, Rx1 ΔpspA, Rx1 pspA1, and Rx1 pspA4 in Todd-Hewitt medium containing 0.5% yeast extract and washed cells in 2% choline chloride (CC). SDS-PAGE analysis of the proteins recovered by a CC wash showed few bands, and the CBPs PspA and PspC (pneumococcal surface protein C) were identified by mass spectrometry analysis. Subcutaneous immunization of mice with these full-length native proteins without adjuvant led to significantly higher rates of survival than immunization with diluent after an intranasal lethal challenge with two pneumococcal strains and also after a colonization challenge with one strain. Importantly, immunization with recombinant PspA4 (rPspA4) without adjuvant did not elicit significant protection.

Vaccine research for gonococcal infections: where are we?

Gonorrhoea continues to seriously impact human society with an estimated 106 million new infections occurring annually. The consequence of gonorrhoea on reproductive and neonatal health is especially concerning as is its role in the spread of HIV. Current control measures rely on the identification and treatment of infected individuals and their sexual contacts. The success of this strategy, which is already inadequate, is lessened by poor diagnostic capabilities in many parts of the world and challenged by the rapid emergence of antibiotic-resistant strains. The potential of untreatable gonorrhoea is now real, and a gonorrhoea vaccine is seriously needed. Historically, gonorrhoea vaccine research has been hampered by the antigenic variability of the gonococcal surface, a lack of known protective mechanisms, and the absence of a small laboratory animal model for testing candidate vaccines and manipulating host responses. Here we discuss recent advances that have rekindled research efforts towards a gonorrhoea vaccine. Several conserved and semiconserved vaccine antigens have been identified that elicit bactericidal antibodies or inhibit target function. A mouse genital tract infection model is available for systematic testing of vaccines, and transgenic mice have been developed to relieve host restrictions. Additionally, several immunological advances have been made including the identification of mechanisms by which Neisseria gonorrhoeae suppresses the adaptive response and the demonstration that Th1 responses clear experimental infection in mice and induce a protective memory response. We also discuss important issues with respect to product development that must be considered when entering the vaccine pipeline.

The concept of "tailor-made", protein-based, outer membrane vesicle vaccines against meningococcal disease.

Protein-based, outer membrane vesicle (OMV) vaccines have previously proven to be efficacious against serogroup B meningococcal disease in Norway and Cuba. Currently, a public health intervention is going on in order to control a serogroup B epidemic in New Zealand. The scale-up and standardization of vaccine production required for controlling the New Zealand epidemic has allowed the establishment of large-scale GMP manufacturing for OMV vaccines. The outcome of this will be licensing of the vaccine in New Zealand and possibly other countries. The availability of licensed OMV vaccines raises the question of whether such vaccines may provide the opportunity to control other outbreaks and epidemics. For instance, such a vaccine could control a localised outbreak of group B meningococci in Normandy, France. "Tailor-made" vaccines, focusing on the sub-capsular antigens may also be considered for use in sub-Saharan Africa for the prevention of the recurrent outbreaks by serogroups A and W135 meningococci. This assumption is based on the epidemiological observation that meningococcal outbreaks in Africa are clonal and are strikingly stable regarding their phenotypic characteristics.

Immunization with bacterial antigens: furunculosis.

Although the nature of the antigens and the immune responses they elicit to achieve immunity to furunculosis are still not well defined, the currently available vaccines comprising A. salmonicida bacterins emulsified in oil adjuvants and delivered by intraperitoneal injection provide remarkably high levels of long-lasting protection. Despite some concern over side-effects, these vaccines have been adopted by most Atlantic salmon farmers over the last four years, transforming a situation where furunculosis outbreaks were becoming catastrophic to one where losses from the disease are negligible. Present evidence indicates that antibody responses to the polysaccharide capsule and iron regulated outer membrane proteins are associated with protection. Furthermore, cell-mediated immune responses involving antigen-induced release of cytokines from lymphocytes and the resultant activation of macrophages with the ability to kill the pathogen are also considered important protective mechanisms. Vaccines comprising whole A. salmonicida cultures grown under iron-restricted conditions and delivered by injection in an oil adjuvant are expected to induce prolonged stimulation of all the above responses. While these vaccines are suitable and effective for administration to salmon smolts there is still a need for mass vaccination by immersion or oral routes for salmonid fry. Effective means of achieving this are still required.

[The protective activity of Yersinia pseudotuberculosis antigens]. / Protektivnaia aktivnost' antigenov Yersinia pseudotuberculosis.

Samples of Y.pseudotuberculosis (serovar I) antigens, represent a high-molecular lipopolysaccharide (LPS) fraction with a mol. wt. of 22.5 kD and fractions of outer membrane proteins isolated by the method of M. Osborn and R. Munson (1974), were tested in comparison with the activity with live cells of Y. pseudotuberculosis I attenuated mutant KV 9/2, having lost its Cad plasmid of virulence with a mol. wt. of 47 MD and carrying 2 attenuating markers: resistance to crystal violet and nalidixic acid. In experiments on guinea pigs pathomorphological studies demonstrated high protective activity of Y.pseudotuberculosis I attenuated mutant KV 9/2 and a pronounced protective effect achieved after the immunization of the animals with complex biopolymers, including a high-molecular LPS fraction and outer membrane proteins.

Lack of protection against gastric Helicobacter infection following immunisation with jack bean urease: the rejection of a novel hypothesis.

The common mucosal immune system was stimulated by oral immunisation with jack bean urease and the adjuvant cholera toxin. A high level of local antibody and serum antibody was induced in mice following hyperimmunisation with this combination. No cross-reacting antibody was found against either Helicobacter pylori or Helicobacter felis. No protection was observed against oral challenge of immunised mice with living H. felis thus disproving the interesting hypothesis of Pallen and Clayton that plant urease might induce a protective immunity against helicobacter infection.

[The development of a technology for the production of live dried vaccines against avian pasteurellosis and swine erysipelas]. / Razrabotka tekhnologii proizvodstva zhivykh sukhikh vaktsin protiv pasterelleza ptits i rozhi svinei.

The technology of the production of dried live vaccine against Pasteurella infection of fowl from Pasteur's 2nd avirulent strain, strains AB and K, has been developed. This technology includes the process of batch cultivation of Pasteurella cells, controlled in such parameters as eH, pO2 and glucose concentration, in fermenters in optimized culture medium, based on Hottinger hydrolysate and fermentative casein-yeast hydrolysate, and preservation in improved saccharose-gelatin medium prepared in potassium sulfate buffer solution. The new technology makes it possible to increase the yield of preparations with stable biological activity 5- to 13-fold in comparison with the traditional technology. Furthermore, the technology of the production of live dried vaccine against swine erysipelas from Erysipelothrix insidiosa strain BP-2 has been developed. This technology is based on maintaining the optimum conditions of the batch cultivation of E. insidiosa in meat medium based on Hottinger hydrolysate and media obtained from hydrolysate of pancreatic fermentation products of microbial biomass; the preparation thus obtained is stabilized in peptone-saccharose-gelatin medium prepared in potassium phosphate buffer solution. This increases the yield of the vaccine 8-fold in comparison with the traditional technology, while ensuring the stability of bacteria after drying and during prolonged storage.

[The action of ribosomal preparations on nonspecific resistance to bacterial infection and on early tolerance to endotoxic shock]. / Deistvie ribosomal'nykh preparatov na nespetsificheskuiu rezistentnost' k bakterial'noi infektsii i ranniuiu tolerantnost' k éndotoksicheskomu shoku.

Ribosomal preparations from Shigella flexneri and Shigella sonnei, introduced parenterally into mice, enhance their resistance to infection with the causative agents of typhoid fever and staphylococci. This effect is considerably less pronounced than that produced by the preparation of homologous lipopolysaccharide isolated by Boivin's method. After the administration of ribosomes nonspecific resistance to bacterial infective agents lasts for a short time. Ribosomal preparations do not enhance the resistance of mice to the lethal action of endotoxin.

[The improvement of immunobiological preparations against leptospirosis. An experimental study of a new concentrated purified vaccine against icterohemorrhagic leptospirosis for human immunization]. / Sovershenstvovanie immunobiologicheskikh protivoleptospiroznykh preparatov. Eksperimental'noe izuchenie novoi kontsentrirovannoi ochishchennoi vaktsiny protiv ikterogemorragicheskogo leptospiroza dlia immunizatsii liudei.

The findings of the study of immunological structure of the population in regions endemic for leptospirosis indicate that the immune status of humans makes it impossible to obtain titrated blood sera for the preparation of antileptospirosis immunoglobulin. The data obtained in the study of the immunobiological properties of a new concentrated vaccine against icterohemorrhagic leptospirosis show the possibility of using this vaccine for the immunization of donors with the aim of obtaining blood sera to be used as raw material for the production of immunobiological preparations.

Immunoprotection of guinea pigs against experimental acute pyelonephritis after the administration of Escherichia coli purified fimbriae.

Experimental results of the present study evidenced the following aspects: a) the antigen, prepared from type I Escherichia coli purified fimbriae (H. 2946 strain), induced immunity at urinary tract level; b) the immunoprotection induced by oral vaccination with multiple doses of fimbriated antigen produced a significant decrease of acute pyelonephritis in newborn guinea pigs and at the same time, a local protection of the urinary tract; c) the immunoprophylaxis by vaccine prepared from fimbriae represents a preferential solution for urinary tract infections prevention in general and especially in children; d) the frequency distribution differences between "protected" and "non-protected" animals were evaluated by chi-square--test with YATES correction and proved to be statistically significant at probability levels.

Investigations on the efficacy of monovalent Pseudomonas aeruginosa vaccine for oral administration in Pseudomonas aeruginosa experimental infection.

Therapeutic efficacy of Pseudomonas aeruginosa vaccine for oral use (10(10) killed germs/ml), prepared from strain 4922, belonging to serotype XV, by Meitert-Meitert scheme, on 4 experimental models in mice (pneumonia, infected burn, septicaemia and urinary tract infection) was studied in comparison with monovalent Ps. aeruginosa vaccine serotype XV (10(9) killed germs/ml) for subcutaneous use and also with associated administration of the two vaccine variants. Mice immunization by using vaccine for oral use was performed by 0.5 ml vaccine per day, for 10 days and vaccine for subcutaneous use was administrated in a volume of 0.5 ml x 2, at 3 days interval. Mice immunization by using the two vaccine types, in association was concomitantly performed and in the same quantity as for separate immunization. In experimental pneumonia, Ps. aeruginosa vaccine for oral use protected mice in 35% of cases, those with infected burns were protected in 33.3% of cases, those with septicemia--in 96.6% of cases and those with urinary tract infection in 50% of cases. As compared to Ps. aeruginosa vaccine for subcutaneous use, the results obtained by vaccine for oral use are less favourable but associated administration of both vaccine variants led to superior results. Thus, in experimental pneumonia, it was obtained a surviving rate of 65% for animals immunized with both vaccine types, in comparison with 50% for animals immunized with vaccine for subcutaneous use only, and in Ps. aeruginosa infected burn, it was obtained a recovering rate of 79.1% for the animals immunized by using both vaccines, in comparison with 70.8% surviving for animals immunized with vaccine for subcutaneous use. In experimental septicaemia and urinary tract infection, combined use of both vaccine variants determined animals surviving and recovering in percents similar to those obtained by separate administration of vaccine for subcutaneous use (in septicemia--100% protection; in urinary tract infection--75% protection).