Protection against bacterial superantigen staphylococcal enterotoxin B by passive vaccination

We investigated the ability of two overlapping fragments of staphylococcal enterotoxin B (SEB), which encompass the whole toxin, to induce protection and also examined if passive transfer of chicken anti-SEB antibodies raised against the holotoxin could protect rhesus monkeys against aerosolized SEB. Although both fragments of SEB were highly immunogenic, the fragments failed to protect mice whether they were injected separately or injected together. Passive transfer of antibody generated in chickens (immunoglobulin Y [IgY]) against the whole toxin suppressed cytokine responses and was protective in mice. All rhesus monkeys treated with the IgY specific for SEB up to 4 h after challenge survived lethal SEB aerosol exposure. These findings suggest that large fragments of SEB may not be ideal for productive vaccination, but passive transfer of SEB-specific antibodies protects nonhuman primates against lethal aerosol challenge. Thus, antibodies raised in chickens against the holotoxin may have potential therapeutic value within a therapeutic window of opportunity after SEB encounter.     

Recombinant C fragment of botulinum neurotoxin B serotype (rBoNTB (HC)) immune response and protection in the rhesus monkey

Botulinum neurotoxin B (BoNTB) is a distinct protein subtype of a family of neurotoxins with the potential for use in biological warfare or terrorist attacks. This study is one in a series evaluating the immunogenicity and protective effects of recombinant vaccines against the different subtypes of botulinum toxin. The recombinant subunit vaccines encoding the C fragment portion (50 kDa) of the toxins are produced in the yeast, Pichia pastoris. In this study, groups of rhesus monkeys were vaccinated with three doses (1 and 5 μg per dose) of rBoNTB(H_c) vaccine. Total and neutralizing antibody titers were determined at various times during and postvaccination. Two groups of vaccinated monkeys plus non-vaccinated controls were actively challenged with B toxin by aerosol exposure. All monkeys receiving vaccine were protected from the toxin and no clinical signs of disease were observed, while controls displaying classic signs of botulism succumbed to the toxin challenge. Two additional groups of monkeys receiving the same vaccine regiment as the first two groups had significant levels of circulating neutralizing antibody titers up to 24 months postvaccination. This non-human primate study demonstrated the short- and long-term immunity afforded by the rBoNTB(H_c) vaccine.     

Correlation of body temperature with protection against staphylococcal enterotoxin B exposure and use in determining vaccine dose-schedule

The immunoprotective potential of a recombinant vaccine against the incapacitating effect of aerosolized staphylococcal enterotoxin B (SEB) in nonhuman primates is reported. SEB belongs to a family of structurally related superantigens responsible for serious, life threatening pathologies. Injecting the recombinant SEB vaccine did not induce temperature elevation in rhesus monkeys, a classical symptom of toxic-shock syndrome. No temperature elevation was noted following injection with control tetanus toxoid. In addition to 100% survival, we observed a clear correlation between vaccine dose and mitigation of temperature elevation after a lethal SEB aerosol challenge. We conclude that the recombinant SEB vaccine is non-pyrogenic and that monitoring changes in body temperature is an important biomarker of toxic shock in a primate animal model.     

Characterization and use of monoclonal and polyclonal antibodies against the mouse interferon-gamma receptor.

To facilitate investigation of its physical and functional properties, 11 monoclonal antibodies (mAbs) and a goat polyclonal IgG specific for the mouse interferon- (IFN-gamma) receptor were characterized and their potential uses studied. Eight of the mAbs interacted with epitopes on the extracellular domain of the receptor, two interacted with epitopes on the intracellular domain, and one interacted with an epitope that could not be localized definitively to either region. Of the 11 mAbs, the majority (8) were IgGs, 2 were IgMs, and 1 was an IgA. Relative avidities of the seven that could be determined ranged from 333 to 0.002 microM-1. Both the polyclonal goat IgG and mAb GR-20 (the latter specific for an epitope in the binding site for IFN-gamma) blocked binding of the ligand and, as expected, prevented induction by IFN-gamma of priming of macrophages for tumor cell killing. None of the other mAbs had an effect despite the fact that GR-22 partially (greater than 50%) blocked binding of IFN-gamma. Neither the polyclonal IgG nor any of the mAbs had an agonist effect. The relative usefulness of the antibodies for immunoprecipitation, immunoblotting, immunoassay, and cell staining with and without prior fixation is described. The results of immunocytochemical staining directly confirmed that the majority of immunologically reactive receptor protein expressed by cells is intracellular. To facilitate use by other investigators, the hybridomas that produce these mAbs will be offered to the American Type Culture Collection.     

Contact sensitizers specifically increase MHC class II expression on murine immature dendritic cells

Contact sensitivity is a T-cell-mediated immune disease that can occur when low-molecular-weight chemicals penetrate the skin. In vivo topical application of chemical sensitizers results in morphological modification of Langerhans cells (LC). Moreover, within 18 h, LC increase their major histocompatibility complex (MHC) class II antigens expression and migrate to lymph nodes where they present the sensitizer to T lymphocytes. We wanted to determine if such an effect could also be observed in vitro. However, because of the high genetic diversity encountered in humans, assays were performed with dendritic cells (DC) obtained from a Balb/c mouse strain. The capacity of a strong sensitizer, DNBS (2,4-dinitrobenzene sulfonic acid), to modulate the phenotype of bone marrow-derived DC in vitro, was investigated. A specific and marked increase of MHC class II molecules expression was observed within 18 h. To eliminate the use of animals in sensitization studies, the XS52 DC line was tested at an immature stage. A 30-min contact with the strong sensitizers DNBS and oxazolone, or the moderate mercaptobenzothiazole, resulted in upregulation of MHC class II molecules expression, analyzed after 18-h incubation. This effect was not observed with irritants (dimethyl sulfoxide and sodium lauryl sulfate) nor with a neutral molecule (sodium chloride). These data suggested the possibility of developing an in vitro model for the identification of the sensitizing potential of chemicals, using a constant and non animal-consuming material.     

Respiratory effects of brevetoxin and saxitoxin in awake guinea pigs

Ptychodiscus brevis toxin (brevetoxin) is associated with 'Florida red tide' and cause neurotoxic shellfish poisoning. Saxitoxin is the agent of paralytic shellfish poisoning. Clinical reports of human intoxication suggest that both toxins affect the respiratory system. The toxins were administered by slow intravenous infusion. The effects of the toxins on respiratory function of awake guinea pigs in a pressure plethysmograph were studied. Both toxins caused lactic acidosis of unknown etiology, which was compensated for by increased minute volume with brevetoxin (PbTx-3)- but not with saxitoxin-intoxicated animals. In general, brevetoxin increased ventilation, before respiratory failure, while saxitoxin had a depressive effect on ventilation. Airways resistance was not increased, nor was dynamic compliance decreased during intoxication, although the data suggest that respiratory system failure was the primary cause of death. The responses seen in these experiments are consistent with the dissimilar molecular actions of these toxins.     

Neonatal consequences of premature rupture of membranes (more than five days) before the 34th week of gestation]

Twenty-six cases of premature braking of the membranes which occurred before week 34 of amenorrhea and lasted for more than 5 days are assessed retrospectively. The mean age when the membranes broke was 26.6 weeks of amenorrhea. Delivery occurred on average at 31.5 +/- 2 WA, with an interval of between 6 and 91 days (mean 35 +/- 23 days). In 4 cases, chorioamniotitis complicated the premature breaking of the membranes. The perinatal mortality rate was 5 out of 27, including 2 still births. Nine of the neonates showed respiratory distress which required artificial ventilation. Four cases of pulmonary hypoplasia were confirmed by pathological examination. In all cases, this was associated with a reduction in the volume of the amniotic fluid, reduced fetal mobility and delayed intrauterine growth. In contrast, when these three factors were absent the prognosis was always good, regardless of the date at which the membranes broke. In the long term, the surviving children showed no neurological sequelae.     

Cardiorespiratory effects of brevetoxin (PbTx-2) in conscious, tethered rats

The cardiorespiratory effects of various doses of brevetoxin (0-100 micrograms PbTx-2/kg) were studied in conscious, tethered rats, After surgical preparation and a 24 hr recovery, toxin or vehicle was infused into the tethered, awake rats for 1 hr. They were then monitored for 6 hr or until death. Toxin-infused rats had decreased core and peripheral temperatures and decreased respiratory rates; these values were all low in the 100 micrograms/kg group at the time of death. Blood gas values remained within normal limits, except terminally. Electrocardiographic (ECG) disturbances, noted in all groups given greater than or equal to 25 micrograms/kg, included heart block, premature ventricular contractions and idioventricular rhythms. It was concluded that brevetoxin causes changes in cardiac conduction and multiple changes in nervous system function.