Showing papers on "Anthrax vaccines published in 2003"

Poly(γ-d-glutamic acid) protein conjugates induce IgG antibodies in mice to the capsule of Bacillus anthracis: A potential addition to the anthrax vaccine

Abstract: Both the protective antigen (PA) and the poly(γ-d-glutamic acid) capsule (γdPGA) are essential for the virulence of Bacillus anthracis. A critical level of vaccine-induced IgG anti-PA confers immunity to anthrax, but there is no information about the protective action of IgG anti-γdPGA. Because the number of spores presented by bioterrorists might be greater than encountered in nature, we sought to induce capsular antibodies to expand the immunity conferred by available anthrax vaccines. The nonimmunogenic γdPGA or corresponding synthetic peptides were bound to BSA, recombinant B. anthracis PA (rPA), or recombinant Pseudomonas aeruginosa exotoxin A (rEPA). To identify the optimal construct, conjugates of B. anthracis γdPGA, Bacillus pumilus γdLPGA, and peptides of varying lengths (5-, 10-, or 20-mers), of the d or l configuration with active groups at the N or C termini, were bound at 5–32 mol per protein. The conjugates were characterized by physico-chemical and immunological assays, including GLC-MS and matrix-assisted laser desorption ionization time-of-flight spectrometry, and immunogenicity in 5- to 6-week-old mice. IgG anti-γdPGA and antiprotein were measured by ELISA. The highest levels of IgG anti-γdPGA were elicited by decamers of γdPGA at 10 –20 mol per protein bound to the N- or C-terminal end. High IgG anti-γdPGA levels were elicited by two injections of 2.5 μg of γdPGA per mouse, whereas three injections were needed to achieve high levels of protein antibodies. rPA was the most effective carrier. Anti-γdPGA induced opsonophagocytic killing of B. anthracis tox–, cap+. γdPGA conjugates may enhance the protection conferred by PA alone. γdPGA-rPA conjugates induced both anti-PA and anti-γdPGA.

Effective Mucosal Immunity to Anthrax: Neutralizing Antibodies and Th Cell Responses Following Nasal Immunization with Protective Antigen

Abstract: Mucosal, but not parenteral, immunization induces immune responses in both systemic and secretory immune compartments. Thus, despite the reports that Abs to the protective Ag of anthrax (PA) have both anti-toxin and anti-spore activities, a vaccine administered parenterally, such as the aluminum-adsorbed anthrax vaccine, will most likely not induce the needed mucosal immunity to efficiently protect the initial site of infection with inhaled anthrax spores. We therefore took a nasal anthrax vaccine approach to attempt to induce protective immunity both at mucosal surfaces and in the peripheral immune compartment. Mice nasally immunized with recombinant PA (rPA) and cholera toxin (CT) as mucosal adjuvant developed high plasma PA-specific IgG Ab responses. Plasma IgA Abs as well as secretory IgA anti-PA Abs in saliva, nasal washes, and fecal extracts were also induced when a higher dose of rPA was used. The anti-PA IgG subclass responses to nasal rPA plus CT consisted of IgG1 and IgG2b Abs. A more balanced profile of IgG subclasses with IgG1, IgG2a, and IgG2b Abs was seen when rPA was given with a CpG oligodeoxynucleotide as adjuvant, suggesting a role for the adjuvants in the nasal rPA-induced immunity. The PA-specific CD4(+) T cells from mice nasally immunized with rPA and CT as adjuvant secreted low levels of CD4(+) Th1-type cytokines in vitro, but exhibited elevated IL-4, IL-5, IL-6, and IL-10 responses. The functional significance of the anti-PA Ab responses was established in an in vitro macrophage toxicity assay in which both plasma and mucosal secretions neutralized the lethal effects of Bacillus anthracis toxin.

The history of anthrax

Abstract: Anthrax, a potentially fatal infection, is a virulent and highly contagious disease. Descriptions of this disease begin in antiquity, with the best ancient account being by the Roman poet Virgil. During the 19th century, anthrax was the infection involved in several important medical developments. It served as the prototype for Koch's postulates regarding the causation of infectious disease. The first vaccine containing attenuated live organisms was Louis Pasteur's veterinary anthrax vaccine. In the 1900s, human inhalation anthrax occurred sporadically in the United States among textile and tanning workers, but the incidence of the illness had declined dramatically. An outbreak of inhalation anthrax occurred in Sverdlovsk near a Soviet military microbiology facility in 1979. This epidemic represented the largest documented outbreak of human inhalation anthrax in history. In October and November 2001, 22 cases of confirmed or suspected inhalation and cutaneous anthrax were reported associated with the intentional release of the organism in the United States. An additional case of cutaneous disease occurred in March of 2002.

A dually active anthrax vaccine that confers protection against both bacilli and toxins.

Abstract: Systemic anthrax is caused by unimpeded bacillar replication and toxin secretion. We developed a dually active anthrax vaccine (DAAV) that confers simultaneous protection against both bacilli and toxins. DAAV was constructed by conjugating capsular poly-γ-d-glutamic acid (PGA) to protective antigen (PA), converting the weakly immunogenic PGA to a potent immunogen, and synergistically enhancing the humoral response to PA. PGA-specific antibodies bound to encapsulated bacilli and promoted the killing of bacilli by complement. PA-specific antibodies neutralized toxin activity and protected immunized mice against lethal challenge with anthrax toxin. Thus, DAAV combines both antibacterial and antitoxic components in a single vaccine against anthrax. DAAV introduces a vaccine design that may be widely applicable against infectious diseases and provides additional tools in medicine and biodefense.

Bacillus anthracis Virulence in Guinea Pigs Vaccinated with Anthrax Vaccine Adsorbed Is Linked to Plasmid Quantities and Clonality

Abstract: Bacillus anthracis is a bacterial pathogen of great importance, both historically and in the present. This study presents data collected from several investigations and indicates that B. anthracis virulence is associated with the clonality and virulence of plasmids pXO1 and pXO2. Guinea pigs vaccinated with Anthrax Vaccine Adsorbed were challenged with 20 B. anthracis isolates representative of worldwide genetic diversity. These same isolates were characterized with respect to plasmid copy number by using a novel method of quantitative PCR developed for rapid and efficient detection of B. anthracis from environmental samples. We found that the copy numbers for both pXO1 and pXO2 differed from those in previously published reports. By combining the data on survival, plasmid copy numbers, and clonality, we developed a model predicting virulence. This model was validated by using a randomly chosen set of 12 additional B. anthracis isolates. Results from this study will be helpful in future efforts to elucidate the basis for variation in the virulence of this important pathogen.

Venezuelan Equine Encephalitis Virus-Vectored Vaccines Protect Mice against Anthrax Spore Challenge

Abstract: Anthrax, a disease usually associated with herbivores, is caused by the bacterium Bacillus anthracis. The current vaccine licensed for human use requires a six-dose primary series and yearly boosters and causes reactogenicity in up to 30% of vaccine recipients. A minimally reactogenic vaccine requiring fewer inoculations is warranted. Venezuelan equine encephalitis (VEE) virus has been configured for use as a vaccine vector for a wide variety of immunogens. The VEE vaccine vector is composed of a self-replicating RNA (replicon) containing all of the VEE virus nonstructural genes and a multiple-cloning site in place of the VEE structural genes. Four different anthrax vaccines were constructed by cloning the protective antigen (PA) gene from B. anthracis into the VEE vaccine vector. The anthrax vaccines were produced by assembling the vectors into propagation-deficient VEE replicon particles in vitro. A/J mice inoculated subcutaneously with three doses of the mature 83-kDa PA vaccine were completely protected from challenge with the Sterne strain of B. anthracis. Similar results were obtained with vaccines composed of the PA gene fused to either the B. anthracis secretory sequence or to a tissue plasminogen activator secretory sequence in three additional mouse strains. Mice were unprotected from challenge after inoculation with the carboxy-terminal 63-kDa PA vaccine. These results suggest that these VEE-vectored vaccines may be suitable as candidate vaccines against anthrax.

Salmonella enterica Serovar Typhimurium Expressing a Chromosomally Integrated Copy of the Bacillus anthracis Protective Antigen Gene Protects Mice against an Anthrax Spore Challenge

Abstract: Protective immunity against infection with Bacillus anthracis is almost entirely based on a response to the protective antigen (PA), the binding moiety for the two other toxin components. We cloned the PA gene into an auxotrophic mutant of Salmonella enterica serovar Typhimurium as a fusion with the signal sequence of the hemolysin (Hly) A gene of Escherichia coli to allow the export of PA via the Hly export system. To stabilize the export cassette, it was also integrated into the chromosome of the live Salmonella carrier. When S. enterica serovar Typhimurium with the chromosomally integrated PA gene was given intravenously to A/J mice, they developed high levels of antibody to PA. These mice were protected against intraperitoneal challenge with 100 or 1,000 50% lethal doses of B. anthracis strain STI. This work contributes to the development of a Salmonella-based orally delivered anthrax vaccine.

Characterisation of the immune response to the UK human anthrax vaccine.

Abstract: The UK human anthrax vaccine consists of the alum-precipitated culture supernatant of Bacillus anthracis Sterne. In addition to protective antigen (PA), the key immunogen, the vaccine also contains a number of other bacteria- and media-derived proteins. These proteins may contribute to the transient side effects experienced by some individuals and could influence the development of the PA-specific immune response. Bacterial cell-wall components have been shown to be potent immunomodulators. B. anthracis expresses two S-layer proteins, EA1 and Sap, which have been demonstrated to be immunogenic in animal studies. These are also immunogenic in man so that convalescent and post-immunisation sera contain specific antibodies to Ea1, and to a lesser extent, to Sap. To determine if these proteins are capable of modifying the protective immune response to PA, A/J mice were immunised with equivalent amounts of recombinant PA and S-layer proteins in the presence of alhydrogel. IgG isotype profiles were determined and the animals were subsequently challenged with spores of B. anthracis STI. The results suggest that there was no significant shift in IgG isotype profile and that the presence of the S-layer proteins did not adversely affect the protective immune response induced by PA.

Serious adverse events among participants in the Centers for Disease Control and Prevention's Anthrax Vaccine and Antimicrobial Availability Program for persons at risk for bioterrorism-related inhalational anthrax.

Abstract: On 20 December 2001, the Centers for Disease Control and Prevention (CDC) initiated the Anthrax Vaccine and Antibiotic Availability Program (hereafter, the "Program") under an investigational new drug application with the US Food and Drug Administration. This Program provided options for additional preventive treatment for persons at risk for inhalation anthrax as a result of recent bioterrorism attacks who had concluded or were concluding a 60-day course of antimicrobial prophylaxis. Participants were offered an additional 40 days of antibiotic therapy (with ciprofloxacin, doxycycline, or amoxicillin) or antibiotic therapy plus 3 doses of anthrax vaccine. By 11 February 2002, a total of 5420 persons had received standardized education about the Program and 1727 persons (32%) had enrolled. Twelve participants have been identified as having serious adverse events (SAEs). One SAE, which occurred in a participant with ciprofloxacin-induced allergic interstitial nephritis, was considered to be probably associated with treatment received in the Program. No SAEs were associated with anthrax vaccine. CDC will continue to monitor Program participants during the next 2 years.

Lymphocytic vasculitis associated with the anthrax vaccine: case report and review of anthrax vaccination.

Abstract: Anthrax is caused by the spore-forming bacteria Bacillus anthracis. It occurs naturally, but recently has been manufactured as a biological warfare agent. This makes prophylaxis for anthrax an urgent concern and efforts are ongoing for the production of an efficient and safe vaccine. Side effects to the current anthrax vaccine are usually minor and mainly consist of local skin reactions. Occasionally an unusual complication may occur; a case of a patient with lymphocytic vasculitis temporally associated with the anthrax vaccine is reported.

Codon-optimized polynucleotide-based vaccines against Bacillus anthracis infection

Abstract: The invention is related to polynucleotide-based anthrax vaccines. In particular, the invention is plasmids operably encoding Bacillus anthracis antigens, in which the naturally-occurring coding regions for the B. anthracis antigens have been modified for improved translation in human or other mammalian cells through codon optimization. In certain embodiments, the coding regions are also modified so as to remove potential N-linked glycosylation sites. B. anthracis antigens which are useful in the invention include, but are not limited to protective antigen (PA), lethal factor (LF), and fragments, variants or derivatives of either of these antigens. The invention is further directed to methods to induce an immune response to B. anthracis in a mammal, for example, a human, comprising delivering a plasmid encoding a codon-optimized B. anthracis antigen as described above. The invention is also directed to pharmaceutical compositions comprising plasmids encoding a codon-optimized B. anthracis antigen as described above, and further comprising adjuvants, excipients, or immune modulators.

Improved anthrax vaccines and delivery methods

Abstract: Immunogenic anthrax compositions, such as anthrax vaccines, and methods for delivering immunogenic anthrax compositions and/or vaccines to a mammal. Methods includes delivery to mucosal membranes and to skin.

Vaccines and bioterrorism: smallpox and anthrax.

Abstract: Because of the success of vaccination and the ring strategy in eradicating smallpox from the world, smallpox vaccine has not been recommended for the United States civilian populations for decades. Given the low but possible threat of bioterrorism, smallpox vaccination is now recommended for those teams investigating potential smallpox cases and for selected personnel of acute-care hospitals who would be needed to care for victims in the event of a terrorist attack. Treatment and post-exposure prophylaxis for anthrax are ciprofloxacin or doxycycline. Anthrax vaccine alone is not effective for post-exposure prevention of anthrax; vaccination is accompanied by 60 days of antibiotic therapy. In addition to military use, anthrax vaccine is recommended for pre-exposure use in those persons whose work involves repeated exposure to Bacillus anthracis spores.

Anthrax refusers: a 2nd infantry division perspective.

Abstract: The Department of Defense anthrax vaccination program has been in the news often recently. Concerns are cited over the safety and usefulness of the vaccine. This brief report describes some of the characteristics of anthrax vaccine refusers. This report examines the implementation of an anthrax vaccination program in a well-disciplined, forward-deployed Army unit facing a hostile enemy with access to anthrax biological warfare stocks.

Molecular characterization of anthrax in positive powders: a Mexican experience.

Abstract: Recently we read contributions from La Scola et al. ([4][1]), Luna et al. ([5][2]), and Bell et al. ([1][3]) published in this journal about their experiences with screening and laboratory analysis carried out to identify anthrax in human samples (nasal swabs) and environmental samples (mail pieces

AIDS research cut to pay for anthrax vaccine.

Abstract: Erika Check,Washington The Bush administration is to proceed with plans to skim more than $200 million from research grant programmes to pay for the rapid production of an anthrax vaccine, brushing off the protests of biologists. As a result of the decision, 375 AIDS researchers and other grant-holders at the National Institute of Allergy and Infectious Diseases (NIAID) will this year lose the initial six months of funding on their awards. The NIAID will spend $233 million on the research, development and purchase of a ‘next-generation’ anthrax vaccine by 2004, the White House Office of Management and Budget says. The transfer of money from the NIAID’s civilian research programmes for the vaccine work is just 2% of its total research budget. But research organizations say they are afraid that the decision is a harbinger of how the NIAID’s swelling biodefence mission may compromise its research programmes. “I don’t think anyone opposes doing research to make a better anthrax vaccine — but that work should be funded out of the bioterrorism budget,not by raiding the AIDS budget,” says Daniel Kuritzkes, director of AIDS research at the Partners AIDS Research Center in Cambridge,Massachusetts. Congress and the administration have been wrangling over the anthrax-vaccine project since February last year, when President Bush’s budget request asked for $233 million for the NIAID to spend on a vaccine. Congress denied the request and divided up the money between several parts of the National Institutes of Health. But the White House then demanded that the NIAID find a way to fulfil its request anyway. In a letter sent to legislators on 2 July, White House budget director Joshua Bolten said that the NIAID would spend up to $117 million this year and $116 million next year on the “advanced development” of an anthrax vaccine, including the purchase of up to 9 million doses of vaccine. The decision disappointed the Infectious Diseases Society of America, which says that the vaccine purchase could endanger the NIAID’s larger research mission. The group argues that another branch of government, such as the Department of Homeland Security, should be paying for the vaccine. On 11 July, Congressman Henry Waxman (Democrat,California) and Senator Jeff Bingaman (Democrat, New Mexico) wrote to the president protesting against the decision,which they called a “serious mistake”. But NIAID officials are putting on a brave face.“The Office of Management and Budget’s position is that there is a critical need for the nation to rapidly develop a vaccine and there’s nothing else out there to support this now,” says Ralph Tate, the NIAID’s budget director. Janet Shoemaker,public-affairs director at the American Society for Microbiology, says NIAID officials are making the best of a difficult situation.“The anthrax issue has become less urgent in most people’s minds, but in the minds of the people making the decisions it is still a very high priority,”she says. ■ At the sharp end: anthrax vaccines for US troops are a high priority for the Bush administration.

Anthrax vaccines and delivery methods

Abstract: Immunogenic anthrax compositions, such as anthrax vaccines, and methods for delivering immunogenic anthrax compositions and/or vaccines to a mammal. Methods include delivery to mucosal membranes and to skin.

Method for complex prophylaxis of anthrax

Abstract: FIELD: medicine, infectious diseases. SUBSTANCE: prophylaxis is carried out by the following schedule: for the first 6 h after infection antibiotic- -resistant mixed anthrax vaccine is administrated subcutaneously as a single human-dose and immunomodulating agent and antibiotic of broad spectrum effect are administrated for 5 days. Invention provides to create preventive protection for a long time against infection with high pathogen doses (100 LD50) from the first day of combined using the complex of agents with specific and nonspecific prophylaxis of anthrax. Invention can be used for complex prophylaxis of anthrax. EFFECT: enhanced effectiveness of method. 4 tbl

Mucosal and systemic Immune Responses Induced with Live Attenuated Anthrax Vaccine A16R in Mice

Abstract: Objective To explore the mucosal find systemic immune mechanisms of live attenuated anthrax vaccine A16R in mice.Methods Divide the BALB/c mice aged 6-8 weeks into 3 groups randomly,5 mice per group. Inoculate s.c. the mice in 2 test groups with live attenuated anthrax vaccine A16R at dosages of 5 x 107 cfu(equal to 1/20 human dose) and 2 x 108 cfu(equal to 1/5 human dose) respectively,and those in control group with 100 ;J normal saline. Collect venous blood every week for detecting the specific IgG to PA in sera by indirect ELISA.Kill the mice 8 weeks after inoculation and separate NALT,NP,spleens,PP nodes and groin lymphocytes to detect cell phenotype by FAGS, and collect lung washings for detecting specific slgA to PA by ELISA. Results IgG were induced in the mice inoculated with the vaccine at 1/5 human dose rapidly and maintained at a high level even 8 weeks after inoculation. However, little slgA was detected in the lung washings of them. No slgA was detected in the lung washings of mice inoculated with 1/20 human dose.No significant difference were observed in the CD3+ , CD4+ , CD8+ and CD45R+ cells in NALT, NP, PP and groin lymph node of the 3 groups. However,compared with those in control group, the CD/ cell constituent ratio in spleen cells of mice in the 2 test groups was low,CD45R+ cell constituent ratio was high,and T/B value was low. No significant difference were observed in the ratios in the 2 test groups. The cells phenotypes in lymph tissue at other sites showed no significant change. Conclusion Anthrax vaccine A16R induced systemic immune response but no effective respiratory or mucosal immune response.

Comparative evaluation of protective antigen produced from Bacillus anthracis & Escherichia coli

Abstract: Interpretation & conclusion: The purified PA preparations obtained in the present study may possibly be utilized for detection of anti-PA antibodies in the sera of anthrax patients for timely diagnosis of the disease and, might also be tested for their efficacy and use as human anthrax vaccine

[Anthrax and carbuncle: two sides of the same coin].

Abstract: The disease caused by Bacillus anthracis is one of the most critical concerns to the general public and public health authorities due both to the anthrax cases caused by the intentional release of the germ in the USA at the close of 2001 when letters and packages were contaminated with anthrax spores, and the current threat of biological warfare. After a brief excursus on the history of the terms Anthrax and Carbuncle, we survey the main evidence of anthrax found in the ancient literature, and deal with the identification of the pathogenic agent responsible for the disease and the subsequent discovery of the first anthrax vaccine and its use in order to control the spread of the disease in the cattle. Finally, we examine some of the most important episodes of occupational exposure to the Bacillus anthracis that occurred in the past two centuries and the preventive measures applied both to employees and the workplace.

Method of preparing native spore suspension for anthrax vaccine preparing

Abstract: FIELD: medicine, microbiology. SUBSTANCE: invention relates to production of anthrax vaccines. Preparing native spore suspension in submerged aerobic culturing vaccine strains of Bacillus anthrax is carried out for two stages: at the first stage culturing is carried out at continuous stirring and aeration with air and maintaining temperature 30-34 C. After termination of spores maturation the second stage is carried out by rise of temperature to 35-38 C and cultural fluid is stirred for short time and periodically. Method allows to increase the content of full-value matured spores in cultural liquid to almost 100%, to determine time of culturing process termination more exactly and to decrease its duration by 2 h. EFFECT: improved method of spore suspension preparing. 3 cl, 2 tbl, 2 ex

Genomics and Proteomics in Vaccine Design

Abstract: In 1881, Louis Pasteur, the father of bacterial vaccines and immunology, demonstrated publicly the first vaccine against a bacterial infection. His vaccine, against anthrax in sheep, consisted of Bacillus anthracis attenuated by high-temperature growth in his laboratory. At Pouilly-Le-Fort, a small village close to Paris, he vaccinated 25 sheep then challenged these plus 25 controls with a virulent strain of B. anthracis. All 25 control sheep died, and all 25 immunized sheep survived. This remarkably successful experiment silenced even his most vocal detractors and paved the way for the development of antibacterial vaccines for use in man. Remarkably, the anthrax vaccine produced today and used to immunize American soldiers is produced in a similar fashion except that a stable partially attenuated strain is used. The second bacterial vaccine was produced by Ramon in 1924 and was essentially a formaldehyde-inactivated supernatant from cultures of Corynebacterium diphtheriae. Again, current vaccines against diphtheria are produced in the same way except that the inactivated toxin that confers protection is partially purified from the culture supernatant. The story continues with the inactivated whole-cell vaccine against Bordetella pertussis (whooping cough) first produced in the late 1940s and still used today in developing countries. It is unlikely that these vaccines would gain FDA approval today. They are still accepted because several decades of use has demonstrated that they are reasonably safe and very effective. It should not be forgotten that these vaccines, together with the smallpox and polio vaccines, essentially eliminated the major causes of childhood mortality in the industrialized world.