Effect of aluminum hydroxide adjuvant and formaldehyde in the formulation of rPA anthrax vaccine

The serological response and efficacy of Bacillus anthracis recombinant protective antigen (rPA) vaccines formulated with aluminum hydroxide adjuvant, either with or without formaldehyde, were evaluated in rabbits. Rabbits that had been injected with a single dose of 25 microg of rPA adsorbed to 500 microg of aluminum in aluminum hydroxide gel (Alhydrogel) had a significantly higher quantitative anti-rPA IgG ELISA titers (p<0.0001) and toxin neutralizing antibody (TNA) assay titers (p<0.0001) than rabbits tested at the next lowest concentration of aluminum (158 microg). Rabbits injected with two doses of 50 microg of rPA formulated with 500 microg of aluminum also had significantly higher serological responses, as measured by a quantitative anti-rPA IgG ELISA (p<0.0001) and TNA assay (p<0.0001), than sera from rabbits injected with a rPA vaccine formulated without adjuvant. Short-term protection against an aerosol spore challenge (448 LD(50)), however, was not significantly different between the two groups (12/12 and 11/12, respectively). Rabbits injected with a single dose of 50 microg of rPA formulated with 500 microg of aluminum and 0.2% formaldehyde had significantly higher ELISA (p<0.0001) and TNA assay (p<0.0001) titers than rabbits that had been injected with a rPA vaccine formulated with adjuvant but without formaldehyde. Short-term protection against a 125 LD(50) parenteral spore challenge, however, was not significantly different between the two groups (14/24 and 9/24, respectively; p=0.2476). Under the conditions tested in the rabbit animal model, significantly higher serological responses were observed in rabbits that had been injected with rPA formulated with aluminum hydroxide gel adjuvant and formaldehyde. However, differences in short-term efficacy were not observed.     

Defining a serological correlate of protection in rabbits for a recombinant anthrax vaccine

In these studies, a serological correlate of protection against anthrax was identified in New Zealand white (NZW) rabbits that had been given one or two injections of various amounts of recombinant protective antigen (rPA) combined with aluminum hydroxide adjuvant (Alhydrogel). Rabbits were subsequently challenged by the aerosol route with spores of the Ames isolate of Bacillus anthracis. Results suggested that the antibody response, as determined by the quantitative anti-rPA IgG ELISA and toxin neutralizing antibody (TNA) assay, were significant predictors (P<0.0015) of protection against a B. anthracis aerosol spore challenge in rabbits.     

A single immunization with a dry powder anthrax vaccine protects rabbits against lethal aerosol challenge

Here we confirm that intranasal (IN) dry powder anthrax vaccine formulations are able to protect rabbits against aerosol challenge 9 weeks after a single immunization. The optimum dose of rPA in our dry powder anthrax vaccine formulation in rabbits was experimentally determined to be 150microg and therefore was chosen as the target dose for all subsequent experiments. Rabbits received a single dose of either 150microg rPA, 150microg rPA+150microg of a conjugated 10-mer peptide representing the Bacillus anthracis capsule (conj), or 150microg of conj alone. All dry powder formulations contained MPL and chitosan (ChiSys). Significant anti-rPA titers and anthrax lethal toxin neutralizing antibody (TNA) levels were seen with both rPA containing vaccines, although rPA-specific IgG and TNA levels were reduced in rabbits immunized with rPA plus conj. Nine weeks after immunization, rabbits were exposed to a mean aerosol challenge dose of 278 LD50 of Ames spores. Groups immunized with rPA or with rPA+conj had significant increases in survivor proportions compared to the negative control group by Logrank test (p=0.0001 and 0.003, respectively), and survival was not statistically different for the rPA and rPA+conj immunized groups (p=0.63). These data demonstrate that a single immunization with our dry powder anthrax vaccine can protect against a lethal aerosol spore challenge 9 weeks later.     

Comparability of ELISA and toxin neutralization to measure immunogenicity of Protective Antigen in mice,as part of a potency test for anthrax vaccines

Complexities of lethal challenge models have prompted the investigation of immunogenicity assays as potency tests of anthrax vaccines. An ELISA and a lethal toxin neutralization assay (TNA) were used to measure antibody response to Protective Antigen (PA) in mice immunized once with either a commercial or a recombinant PA (rPA) vaccine formulated in-house. Even though ELISA and TNA results showed correlation, ELISA results may not be able to accurately predict TNA results in this single immunization model.     

Temperature-mediated recombinant anthrax protective antigen aggregate development: Implications for toxin formation and immunogenicity

Anthrax vaccines containing recombinant PA (rPA) as the only antigen face a stability issue: rPA forms aggregates in solution after exposure to temperatures ⩾40 °C, thus losing its ability to form lethal toxin (LeTx) with Lethal Factor. To study rPA aggregation’s impact on immune response, we subjected rPA to several time and temperature combinations. rPA treated at 50 °C for 30 min formed high mass aggregates when analyzed by gel electrophoresis and failed to form LeTx as measured by a macrophage lysis assay (MLA). Aggregated rPA-formed LeTx was about 30 times less active than LeTx containing native rPA. Mice immunized with heat-treated rPA combined with Al(OH)₃ developed antibody titers about 49 times lower than mice immunized with native rPA, as measured by a Toxicity Neutralization Assay (TNA). Enzyme Linked Immunosorbent Assay (ELISA) of the same immune sera showed anti-rPA titers only 2–7 times lower than titers elicited by native rPA. Thus, rPA’s ability to form LeTx correlates with its production of neutralizing antibodies, and aggregation significantly impairs the protein’s antibody response. However, while these findings suggest MLA has some value as an in-process quality test for rPA in new anthrax vaccines, they also confirm the superiority of TNA for use in vaccine potency.     

Immunogenicity and safety of a novel recombinant protective antigen anthrax vaccine (GC1109), a randomized,single-blind,placebo controlled phase II clinical study

BackgroundThe demand on effective and safe anthrax vaccine is increasing as a part of the preparedness for possible bioterrorism in the future. We performed a randomized, single-blind, placebo controlled phase II clinical study to evaluate the immunogenicity and safety of a novel recombinant protective antigen (rPA) anthrax vaccine, GC1109, in healthy adult volunteers.MethodsParticipants were randomized to experiment groups (0.3 mL, 0.5 mL, and 1.0 mL of GC1109) or placebo group (normal saline 0.5 mL) in 2:2:2:1 ratio. They received respective vaccines intramuscularly at 0, 4 and 8 weeks. Immunogenicity was evaluated by seroconversion rate and geometric mean titer (GMT) of lethal toxin neutralizing assay (TNA) and anti-PA IgG by ELISA. Safety was assessed by laboratory tests, and solicited and unsolicited adverse events on diary cards.Results30, 29, 30 participants were randomized to 0.3, 0.5, and 1.0 mL of GC1109 groups, respectively, while 15 to placebo group. 92 participants received all three doses. In per-protocol analysis, TNA GMTs at week 12 were 296.5, 285.2, and 433.2 in the three groups, respectively. Seroconversion rates measured by ELISA were 100% at week 12 in the three groups. Local and systemic vaccine-related adverse events were frequent; however, most of them were mild, and no serious events were observed.ConclusionsA new rPA anthrax vaccine GC1109 was immunogenic after three doses of intramuscular administration, and was well-tolerated.     

Development of an in vitro-based potency assay for anthrax vaccine

The potency assay currently used to evaluate consistency of manufacture for the anthrax vaccine is contingent upon meeting specified parameters after statistical analysis of the percent survival and time to death of vaccinated guinea pigs after challenge with spores of a virulent strain of Bacillus anthracis. During the development of a new anthrax vaccine based upon recombinant protective antigen (rPA) adsorbed to aluminum hydroxide gel (Alhydrogel), we found that the serological response of female A/J mice, as measured by a quantitative anti-rPA IgG ELISA, may be an effective method to monitor a manufacturer's consistency for rPA-based vaccines. An advantage of the proposed in vitro-based potency assay is that it will not need stringent biosafety containment measures as required by the current guinea pig potency assay.     

Immunogenicity and tolerance of ascending doses of a recombinant protective antigen (rPA102) anthrax vaccine: a randomized, double-blinded, controlled, multicenter trial

BACKGROUND: We report the results of a phase I dose escalation, safety and immunogenicity trial of a new recombinant protective antigen (rPA102) anthrax vaccine. METHODS: Hundred healthy volunteers were randomized in a 4:1 ratio to receive intramuscular doses of rPA102 in the following formulations: 5, 25, 50, or 75 microg of rPA102 in 82.5 microg aluminum hydroxide adjuvant at 0, 4, and 8 weeks; or the US licensed Anthrax Vaccine Adsorbed (AVA) at weeks 0 and 4. FINDINGS: Local reactogenicity (mostly pain) was more common with AVA than with rPA102 following the first (94.7% versus 44.4%; p < 0.001) and the second (84.2% versus 35.4%; p < 0.001) vaccinations. Systemic reactogenicity (mostly headache) was more common among rPA102 vaccinees, but only following the first vaccination (49.4% versus 15.8%; p = 0.025). A dose-response relationship for anti-PA antibodies was present after the 2nd and 3rd vaccinations. Two weeks following the 2nd vaccination, the geometric mean titers (GMT) for lethal toxin neutralization activity (TNA), for the 5, 25, 50 and 75 microg rPA102 and AVA groups were 38.6, 75.4, 373.9, 515.3, and 855.2, respectively. The geometric mean concentrations (GMC) measured by anti-PA IgG ELISA were 3.7, 11.5, 25.9, 44.1, and 171.6, respectively. Two weeks following the 3rd vaccination, TNA GMTs for the four rPA102 groups, were: 134.7, 719.7, 2116.6, 2422.4; and ELISA GMCs were: 22.9, 104.7, 196.4, and 262.6, respectively. INTERPRETATION: No clinically serious or dose-related toxicity or reactogenicity was observed. The TNA response after two injections of the 75 microg dose of rPA102 was similar to the response after two injections of AVA. The third rPA102 vaccination substantially increased the antibody response.     

Serological Correlate of Protection in Guinea Pigs for a Recombinant Protective Antigen Anthrax Vaccine Produced from Bacillus brevis

ObjectiveRecombinant protective antigen (rPA) is the active pharmaceutical ingredient of a second generation anthrax vaccine undergoing clinical trials both in Korea and the USA. By using the rPA produced from Bacillus brevis pNU212 expression system, correlations of serological immune response to anthrax protection efficacy were analyzed in a guinea pig model.MethodsSerological responses of rPA anthrax vaccine were investigated in guinea pigs that were given single or two injections (interval of 4 weeks) of various amounts of rPA combined with aluminum hydroxide adjuvant. Guinea pigs were subsequently challenged by the intramuscular injection with 30 half-lethal doses (30LD₅₀) of virulent Bacillus anthracis spores. Serum antibody titers were determined by anti-PA IgG ELISA and the ability of antibodies to neutralize the cytotoxicity of lethal toxin on J774A.1 cell was measured through the toxin neutralizing antibody (TNA) assay.ResultsTo examine correlations between survival rate and antibody titers, correlation between neutralizing antibody titers and the extent of protection was determined. Toxin neutralization titers of at least 1176 were sufficient to confer protection against a dose of 30LD₅₀ of virulent anthrax spores of the H9401 strain. Such consistency in the correlation was not observed from those antibody titers determined by ELISA.ConclusionNeutralizing-antibody titers can be used as a surrogate marker.     

Protective activity and immunogenicity of two recombinant anthrax vaccines for veterinary use

In this study, the efficacy of two experimental vaccines against Bacillus anthracis toxinaemia was evaluated in the rabbit model. A recombinant Protective Antigen (rPA) mutant and a trivalent vaccine (TV) composed by the rPA, a inactive mutant of Lethal Factor (mLF-Y728A; E735A) and a inactive mutant of Edema Factor (mEF-K346R), both emulsified with mineral oils, were evaluated for their immunogenicity and protective activity in New Zealand white rabbits. Rabbits vaccinated subcutaneously with rPA and TV rapidly produced high level of anti-PA, anti-LF and anti-EF antibodies, which were still present 6 months later. In the efficacy test, these vaccines protected 100% of rabbits challenged with B. anthracis virulent strain 0843 one week after the vaccination. Moreover, all animals vaccinated twice with rPA and TV, resisted B. anthracis infection 6 months later. Our data indicate that rPA and TV could be good vaccine candidates for inducing protection against B. anthracis infection in target animal host. They could successfully be used in an emergency with simultaneous long-acting antibiotics to halt incubating infections or during an anthrax epidemic.     

Evaluation of the protective efficacy of recombinant protective antigen vaccine (GC1109)-immunized human sera using passive immunization in a mouse model

The protective efficacy of human sera from vaccinated individuals with a new recombinant protective antigen anthrax vaccine (GC1109) against lethal spore challenge was evaluated in a mouse model. Eighteen human sera were selected from the vaccinated individuals based on their toxin neutralizing assay (TNA) titer (ED₅₀ of 55 to 668). The selected sera were diluted and passively transferred to A/J mice and the mice were subsequently challenged with 100 × LD₅₀ of Bacillus anthracis Sterne spores. The correlation between the survival rate of passively immunized mice and the TNA ED₅₀ of transferred sera was presented (r = 0.873, P-value < 0.001). The estimated TNA titer for 50% survival rate against lethal challenge was 197 (95% confidence interval of 149 and 260). The result suggest that GC1109 is protective against exposure to B. anthracis and the TNA titer of vaccinated serum can be an indicator for protective efficacy.     

Attenuated immune response to tetanus toxoid in young healthy men protected against tetanus

Tetanus booster is a routine procedure of tetanus prevention in populations with high risk of injury, independent of the levels of protection. But the immune response in already protected individuals is not well studied. We describe the kinetics of booster response in individuals by measuring tetanus antitoxin levels by indirect ELISA. A 6-month follow up was performed on 60 boosted individuals tested before, 1 week, 1, 2, 3 and 6 months after the booster. High initial protection (mean titer 1.08 IU/ml) and less than 3-fold increase after 1 month were observed. After 1 month of stable antitoxin levels, the levels slowly decreased and reached a mean titer of 1.78 IU/ml after 6 months. Individuals with initial levels <1 IU/ml had booster response after the first month twice as high compared to those with initial level >or=1 IU/ml. However, in both groups, the decline from 1 to 6 months was about 2-fold. Individuals already protected against tetanus exhibited an attenuated, short-lasting booster response to tetanus toxoid. This was more pronounced in individuals with pre-booster levels >or=1 IU/ml, who did not improve immune protection after the booster.     

Immunogenicity in mice of anthrax recombinant protective antigen in the presence of aluminum adjuvants

The only US-licensed anthrax vaccine for human use, as well as several experimental vaccines containing solely purified recombinant protective antigen (rPA), are formulated using aluminum hydroxide (Al(OH)3) as an adjuvant. It has been suggested that effective adjuvanticity of aluminum salts for protein antigens depends, at least partially, on the degree of adsorption of the antigen to the adjuvant. On the other hand, the ease of antigen desorption from the adjuvant in a quantitative fashion may facilitate the assessment of vaccine characteristics in the laboratory. In this regard, aluminum phosphate (AlPO4), although deemed a "weaker" adjuvant than Al(OH)3, appears superior to the latter. To investigate the possibility of formulating rPA vaccines with AlPO4, as well as the significance of the adsorption of this antigen to the aluminum salt for adjuvanticity, we studied the effect of AlPO4 and Al(OH)3 on the induction of anti-rPA antibodies in mice. In a first immunization experiment the adjuvanticity of AlPO4 combined with rPA was examined. Antibodies against rPA were measured using an ELISA. Results indicated that AlPO4 is able to significantly increase the antibody response to rPA, irrespective of its degree of adsorption to the adjuvant. Based on these results, in a second experiment mice were immunized twice, with different formulations of rPA containing either AlPO4 or Al(OH)3, and rPA-antibodies were measured using ELISA and an in vitro toxin neutralization assay. Comparable immune responses to rPA were obtained with both aluminum salts. Additionally, results with AlPO4 as adjuvant confirmed that, in this mouse model, binding of the protein to the adjuvant is not essential for adjuvanticity, whereas the amount of adjuvant has an influence on the antibody response induced.     

Immunogenicity and Protective Efficacy of Recombinant Protective Antigen Anthrax Vaccine (GC1109) in A/J Mice Model

A recombinant protective antigen anthrax vaccine (GC1109) is being developed as a new-generation vaccine by the Korea Disease Control and Prevention Agency. In accordance with the ongoing step 2 of phase II clinical trials, the immunogenicity and protective efficacy of the booster dose of GC1109 were evaluated in A/J mice after 3 serial vaccinations at 4-week intervals. The results indicated that the booster dose significantly increased the production of anti-protective antigen (PA) IgG and toxin-neutralizing antibody (TNA) compared with those of the group without booster. An enhanced protective effect of the booster dose was not observed because the TNA titers of the group without booster were high enough to confer protection against spore challenge. Additionally, the correlation between TNA titers and probability of survival was determined for calculating the threshold TNA titer levels associated with protection. The threshold 50 % neutralization factor (NF₅₀) of TNA showing 70 % probability of protection was 0.21 in A/J mice with 1,200 LD₅₀ Sterne spores challenge. These results indicate that GC1109 is a promising candidate as a new-generation anthrax vaccine and that a booster dose might provide enhanced protection by producing toxin-neutralizing antibodies.     

A recombinant 63-kDa form of Bacillus anthracis protective antigen produced in the yeast Saccharomyces cerevisiae provides protection in rabbit and primate inhalational challenge models of anthrax infection

Infection by Bacillus anthracis is preventable by prophylactic vaccination with several naturally derived and recombinant vaccine preparations. Existing data suggests that protection is mediated by antibodies directed against the protective antigen (PA) component of the anthrax toxin complex. PA is an 83-kDa protein cleaved in vivo to yield a biologically active 63-kDa protein. In an effort to evaluate the potential of yeast as an expression system for the production of recombinant PA, and to determine if the yeast-purified rPA63 can protect from a lethal inhalational challenge, the sequence of the 63-kDa form of PA was codon-optimized and expressed in the yeast Saccharomyces cerevisiae. Highly purified rPA63 isolated from Saccharomyces under denaturing conditions demonstrated reduced biological activity in a macrophage-killing assay compared to non-denatured rPA83 purified from Escherichia coli. Rabbits and non-human primates (NHP) immunized with rPA63 and later challenged with a lethal dose of B. anthracis spores were generally protected from infection. These results indicate that epitopes present in the 63-kDa from of PA can protect rabbits and non-human primates from a lethal spore challenge, and further suggest that a fully functional rPA63 is not required in order to provide these epitopes.     

Experimental infection of rabbits with Fasciola gigantica and treatment with triclabendazole (TCBZ) or BT6: parasitological, haematological and immunological findings

The responses of rabbits experimentally infected orally with 25 Fasciola gigantica metacercariae each, to treatment with triclabendazole (TCBZ) in a single oral dose of 10 mg/Kg body weight or with the newly synthesized drug (BT6) in repeated oral doses of 500 mg/Kg for 3 consecutive days, were monitored by faecal egg and fluke counts and the enzyme-linked immunosorbent assay (ELISA), both treatments at 4-, 8- and 12- weeks of infection. In all infected rabbits, eggs appeared in the faeces between 10 and 16 weeks post-infection (P1) and Were egg-negative in the TCBZ-treated animals. Compared with the infected untreated control (group B), the reduction in the fluke burdens were 93.75,100 and 100% in TCBZ-treated rabbits (groups C, D and E), respectively and 33.33, 27.08 and 27.08% in the BT6 treated groups F, G and H, respectively. Total red blood corpuscle (RBCs) counts, haemoglobin (Hb) concentration and packed-cell volume (PCV) values in rabbits (group B) indicated the presence of moderate anaemia from the start of infection with a significant decrease from 10-12 weeks onwards. RBCs count, Hb and PCV values from groups C, D and E were significantly higher (p < 0.001) than in those from group B on week 10, 8 and 10 PI, respectively. The increasing was continued till week 16 PI. By two weeks PI there was significant eosinophilia in infected rabbits, maintained at 4 and 6 weeks PI then dropped up to 8 weeks but still statistically higher than the control values. The ELISA detected antibodies against F. gigantica as early as week 2 PI, rising to high levels 6, 10 and 12 weeks later in groups C, D and E, respectively, whilst peaked at 10 weeks in BT6 treated animals, then tended to drop but remained positive through 16 weeks. We conclude TCBZ is a potent fasciolicidic drug. Eosinophilia and anaemia might be indicators for fascioliasis. The ELISA with adult fluke excretory - secretory (ES) antigens could be a feasible method for the diagnosis of experimental fascioliasis in rabbits and post-treatment monitoring.     

Bikes for Life: Measuring the effects of a bicycle distribution program on 6 to 12-year-old children’s BMI and health behaviors

Treating pediatric obesity is challenging. The objective was to evaluate effect of receiving a bicycle on (a) physical activity, (b) sedentary activity, (c) Body Mass Index (BMI), and (d) eating habits.A stepped-wedge randomized controlled trial of 6- to 12-year-old patients with overweight/obesity was conducted April 2012–2018. Participants were randomized to wait 0, 2, 4, or 6 months for a bicycle. Outcomes on activity, BMI and eating were collected at 3, 6, 9- and 12-months after children received a bicycle.A total of 525 participants with 387 (74%) completed 3-month follow-up questionnaire, and 346 (66%) completed 12-month follow-up visit. Participants were mostly Latino/a (71%) and low income (58%), and 31% had never ridden a bicycle. Median baseline BMI was 98th percentile. At 3 months, 62% reported bicycle use last week, on average 3.6 days. Time spent on sedentary activities decreased by 48 min/day (p = 0.04), and time spent playing sports increased by 1.7 h/week (p < 0.01). No reduction in BMI was seen. Consumption of sugary drinks decreased (by 0.59 servings/week, p < 0.01), and consumption of vegetables increased (0.71 servings/week, p = 0.04). At 12 months, sedentary time, sugary drink and vegetable consumption remained significantly more favorable than at enrollment (p < 0.01, p < 0.01, p = 0.04 respectively), but not significantly different (p = 0.47 for sedentary, p = 0.73 for sugary drink) and significantly less favorable (p < 0.01 for vegetables) than at the time of intervention.Participants reported riding bicycle, improved activity and dietary habits, though reversion towards baseline behavior was seen by one year and no change in BMI from enrollment.     

Passive immunotherapy of Bacillus anthracis pulmonary infection in mice with antisera produced by DNA immunization

Because of the high failure rate of antibiotic treatment in patients with anthrax there is a need for additional therapies such as passive immunization with therapeutic antibodies. In this study, we used codon-optimized plasmid DNAs (DNA vaccines) encoding Bacillus anthracis protective antigen (PA) to immunize rabbits for producing anti-anthrax antibodies for use in passive immunotherapy. The antisera generated with these DNA vaccines were of high titer as measured by ELISA. The antisera were also able to protect J774 macrophage cells by neutralizing the cytotoxic effect of exogenously added anthrax lethal toxin, and of the toxin released by B. anthracis (Sterne strain) spores following infection. In addition, the antisera passively protected mice against pulmonary challenge with an approximate 50 LD50 dose of B. anthracis (Sterne strain) spores. The protection in mice was obtained when the antiserum was given 1h before or 1h after challenge. We further demonstrated that IgG and F(ab')2 components purified from anti-PA rabbit hyperimmune sera retained similar levels of neutralizing activities against both exogenously added B. anthracis lethal toxin and toxin produced by B. anthracis (Sterne strain) spores. The high titer antisera we produced will enable an immunization strategy to supplement antibiotic therapy for improving the survival of patients with anthrax.     

Promoting and maintaining physical activity in people with type 2 diabetes

BACKGROUND: Limited research has investigated how to promote physical activity in people with type 2 diabetes. This study evaluated physical activity counseling over 12 months in people with type 2 diabetes. DESIGN: Participants were given standard exercise information and randomly assigned to receive physical activity counseling or not. Data were collected from September 2000 through to September 2002 and analyzed from October 2002 to February 2003. SETTING/PARTICIPANTS: Diabetes outpatient clinic. Seventy inactive people with type 2 diabetes. INTERVENTION: Physical activity counseling, based on the transtheoretical model, combined motivational theory and cognitive behavioral strategies into an individualized intervention to promote physical activity. Consultations were delivered at baseline and 6 months, with phone calls at 1 and 3 months post-consultation. MAIN OUTCOME MEASURES: Changes from baseline at 12 months in physical activity (7-day recall and accelerometer), stages and processes of exercise behavior change. RESULTS: Between-group differences were recorded in physical activity (recall and accelerometer) at 12 months (p <0.01). Experimental participants significantly increased total activity (median difference, 115 minutes; 95% confidence interval [CI]=73-150 minutes). Control participants recorded no significant change (median difference, -15 minutes; 95% CI=-53-13 minutes). The accelerometer experimental participants recorded no significant change (mean difference, 416,632 counts; 95% CI=-27,743, 1,051,007 counts/week), while control participants recorded a significant decrease (mean difference, -669,061 counts; 95% CI=-1,292,285, -45,837 counts/week). At 12 months, more experimental participants compared to controls were in active stages of behavior(6-month chi(2)=26.4, p <0.01; 12-month chi(2)=19.9, p <0.01, respectively). Between-group differences were recorded at 12 months for the frequency of using all processes (p <0.01), except dramatic relief and stimulus control. CONCLUSIONS: Physical activity counseling was effective for promoting physical activity over 12 months in people with type 2 diabetes.     

Correlation between anthrax lethal toxin neutralizing antibody levels and survival in guinea pigs and nonhuman primates vaccinated with the AV7909 anthrax vaccine candidate

The anthrax vaccine candidate AV7909 is being developed as a next generation vaccine for a post-exposure prophylaxis (PEP) indication against anthrax. AV7909 consists of the Anthrax Vaccine Adsorbed (AVA, BioThrax®) bulk drug substance adjuvanted with the immunostimulatory oligodeoxynucleotide (ODN) compound, CPG 7909. The addition of CPG 7909 to AVA enhances both the magnitude and the kinetics of antibody responses in animals and human subjects, making AV7909 a suitable next-generation vaccine for use in a PEP setting. The studies described here provide initial information on AV7909-induced toxin-neutralizing antibody (TNA) levels associated with the protection of animals from lethal Bacillus anthracis challenge. Guinea pigs or nonhuman primates (NHPs) were immunized on Days 0 and 28 with various dilutions of AV7909, AVA or a saline or Alhydrogel + CPG 7909 control. Animals were challenged via the inhalational route with a lethal dose of aerosolized B. anthracis (Ames strain) spores and observed for clinical signs of disease and mortality. The relationship between pre-challenge serum TNA levels and survival following challenge was determined in order to calculate a threshold TNA level associated with protection. Immunisation with AV7909 induced a rapid, highly protective TNA response in guinea pigs and NHPs. Surprisingly, the TNA threshold associated with a 70% probability of survival for AV7909 immunized animals was substantially lower than the threshold which has been established for the licensed AVA vaccine. The results of this study suggest that the TNA threshold of protection against anthrax could be modified by the addition of an immune stimulant such as CPG 7909 and that the TNA levels associated with protection may be vaccine-specific.