A cationic lipid-formulated plasmid DNA vaccine confers sustained antibody-mediated protection against aerosolized anthrax spores.

DNA vaccines provide an attractive technology platform against bioterrorism agents due to their safety record in humans and ease of construction, testing, and manufacture. We have designed monovalent and bivalent anthrax plasmid DNA (pDNA) vaccines encoding genetically detoxified protective antigen (PA) and lethal factor (LF) proteins and tested their immunogenicity and ability to protect rabbits from an aerosolized inhalation spore challenge. Immune responses after two or three injections of cationic lipid-formulated PA, PA plus LF, or LF pDNAs were at least equivalent to two doses of anthrax vaccine adsorbed (AVA). High titers of anti-PA, anti-LF, and neutralizing antibody to lethal toxin (Letx) were achieved in all rabbits. Eight or nine animals in each group were challenged with 100x LD(50) of aerosolized anthrax spores 5 or 9 weeks after vaccination. An additional 10 animals vaccinated with PA pDNA were challenged >7 months postvaccination. All animals receiving PA or PA plus LF pDNA vaccines were protected. In addition, 5 of 9 animals receiving LF pDNA survived, and the time to death was significantly delayed in the others. Groups receiving three immunizations with PA or PA plus LF pDNA showed no increase in anti-PA, anti-LF, or Letx neutralizing antibody titers postchallenge, suggesting little or no spore germination. In contrast, titer increases were seen in AVA animals, and in surviving animals vaccinated with LF pDNA alone. Preclinical evaluation of this cationic lipid-formulated bivalent PA and LF vaccine is complete, and the vaccine has received U.S. Food and Drug Administration Investigational New Drug allowance.

Specificity of an immunochromatographic test for anthrax.

OBJECTIVE: To evaluate the specificity of an immunochromatographic test (ICT) for anthrax in cattle. DESIGN: A comparison of an ICT with blood smear and culture in uninfected cattle. PROCEDURE: Two hundred and forty blood samples were collected from dead cattle at two knackeries within Victoria and tested on-site with an ICT for the detection of protective antigen (PA) of Bacillus anthracis. Blood smears were prepared on-site and blood samples transported to the laboratory for aerobic and anaerobic culture. The results of the ICT were compared with blood smear and culture. Animals were regarded as not infected with B anthracis if the organism was not detected in a stained blood smear or on culture. Ten healthy yearling cattle were vaccinated with live spore anthrax vaccine and blood samples collected on days 0 to 7 and day 15 were tested in the ICT for the presence of PA. RESULTS: All blood samples from the 240 knackery cattle were ICT, smear and culture negative. All blood samples from the 10 vaccinated cattle were ICT negative. CONCLUSION: The ICT is a test with high specificity in cattle (98.5 to 100%; 95% CI) and recent vaccination of cattle does not give rise to positive reactions.

Protection against anthrax lethal toxin challenge by genetic immunization with a plasmid encoding the lethal factor protein.

The ability of genetic vaccination to protect against a lethal challenge of anthrax toxin was evaluated. BALB/c mice were immunized via gene gun inoculation with eucaryotic expression vector plasmids encoding either a fragment of the protective antigen (PA) or a fragment of lethal factor (LF). Plasmid pCLF4 contains the N-terminal region (amino acids [aa] 10 to 254) of Bacillus anthracis LF cloned into the pCI expression plasmid. Plasmid pCPA contains a biologically active portion (aa 175 to 764) of B. anthracis PA cloned into the pCI expression vector. One-micrometer-diameter gold particles were coated with plasmid pCLF4 or pCPA or a 1:1 mixture of both and injected into mice via gene gun (1 microg of plasmid DNA/injection) three times at 2-week intervals. Sera were collected and analyzed for antibody titer as well as antibody isotype. Significantly, titers of antibody to both PA and LF from mice immunized with the combination of pCPA and pCLF4 were four to five times greater than titers from mice immunized with either gene alone. Two weeks following the third and final plasmid DNA boost, all mice were challenged with 5 50% lethal doses of lethal toxin (PA plus LF) injected intravenously into the tail vein. All mice immunized with pCLF4, pCPA, or the combination of both survived the challenge, whereas all unimmunized mice did not survive. These results demonstrate that DNA-based immunization alone can provide protection against a lethal toxin challenge and that DNA immunization against the LF antigen alone provides complete protection.

Azithromycin compared with rifampin for eradication of nasopharyngeal colonization by Neisseria meningitidis.

OBJECTIVES: To evaluate the efficacy and safety of azithromycin compared with rifampin for eradication of nasopharyngeal carriage of Neisseria meningitidis METHODS: Pharyngeal swabs were obtained from 500 students attending nursing school in Cairo, Egypt, to determine the colonization rate with N. meningitidis. Colonized individuals were randomized to receive azithromycin (500 mg once) or rifampin (600 mg twice daily for four doses). Subjects were then recultured 1 and 2 weeks posttreatment to determine the effectiveness of the antibiotic therapy for eradication of meningococcal nasopharyngeal colonization. RESULTS: Individuals treated with azithromycin had a 93% eradication rate at 1 and 2 weeks posttreatment comparable with 95 and 91%, respectively, for rifampin. No significant side effects were reported by any subjects treated with either antibiotic. CONCLUSION: Azithromycin is effective in the eradication of N. meningitidis from the nasopharynx of asymptomatic colonized individuals and deserves further evaluation for use as prophylaxis against N. meningitidis.

Tuberculosis meningitis, Abbassia Fever Hospital-Naval Medical Research Unit No. 3-Cairo, Egypt, from 1976 to 1996.

A total of 1,430 patients with the presumptive diagnosis of tuberculous meningitis were admitted to the U.S. Naval Medical Research Unit No. 3/Abbassia Fever Hospital in Cairo, Egypt from January 1976 to January 1996. Diagnosis was confirmed by culture of the mycobacteria from the cerebrospinal fluid CSF of 857 patients and these patients are included in the final analysis. There were 497 males and 360 females. The patients ranged in age from five months to 55 years. The number of patients admitted during the months of March, April, and May were more than double those admitted during October, November, and December. The duration of symptoms prior to admission ranged from seven to 90 days (mean = 29.5 days). Upon admission, 4% of the patients were alert, 34% were drowsy, and 62% were in a coma. Of the 857 patients studied, 490 (57%) died, 256 (30%) recovered completely, and 11 (13%) recovered with sequelae. The mortality and neurologic sequelae were directly related to the stage of disease and duration of symptoms prior to admission. Mortality was significantly lower in patients admitted in stage II and or with short duration of disease compared with those in stage III and or with prolonged duration of symptoms prior to admission. The use of dexamethasone in patients with tuberculous meningitis significantly reduced the ocular complications that occur in these patients and also significantly reduced the fatality rate.

Focal lesions of the spleen in patients with fever of unknown origin: sonographic patterns and diagnosis.

A review of ultrasonic scans performed for 811 patients admitted at the Abbassia Fever Hospital between 1990 through 1996 for investigation of fever undiagnosed for over 3 weeks revealed 28 cases of focal splenic lesions. They included lymphoma in sixteen cases, tuberculosis in six cases, echinococcal cysts in three, abscess in two, and infarction in one. Final diagnosis was obtained by correlating the ultrasonography and clinical findings, together with pathologic, bacteriologic, serologic and angiographic data.

Splenic tuberculosis in patients with fever of unknown origin.

Splenic tuberculosis is uncommon. We report here splenic tuberculosis in 5 patients with fever of over 3 weeks duration. In the 5 patients, abdominal ultrasonography and/or abdominal computed tomography revealed multiple hypoechoic and hypodense splenic lesions. Diagnosis required biopsy of cervical lymph nodes in four cases and splenectomy in one. Despite vigorous antituberculous therapy with isoniazid, rifampin, and ethambutol, the clinical condition of 2 of these patients worsened and splenectomy was done. Histological examination of a tissue specimen of the spleen showed multiple caseating granulomas.

The effect of varying time at -Gz on subsequent +Gz physiological tolerance (push-pull effect).

INTRODUCTION: Previous studies have demonstrated decreased +Gz tolerance when preceded by 0 Gz or -Gz, referred to as the "push-pull effect." The purpose of this experiment was to observe the effect of varying time duration at -Gz on the push-pull effect. METHODS: During single sessions, six subjects (three men, three women) were subjected to five relaxed exposures to +2.25 Gz on the NAMRL Coriolis Acceleration Platform (CAP). The first and last exposures were control runs that were preceded by +1 Gz. Each experimental run was preceded by -2 Gz for 2, 5, or 15 s. Blood pressure (BP) was monitored using the Finapres at the level of the clavicle. Visual light loss was assessed at +2.25 Gz using a light bar. RESULTS: Mean BP was significantly reduced when the +2.25 Gz exposures were preceded by -2 Gz. Following 15 s of -2 Gz, mean BP decreased more and was slower to recover than for 2 and 5 s of -2 Gz. Reported incidents of visual light loss were: 1 following 2 s, 2 following 5 s, and 4 following 15 s at -2 Gz. There were no reports of visual light loss during control runs. CONCLUSION: During relaxed conditions, the push-pull effect is augmented by increasing duration of the preceding -Gz.