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PubMed1995

Tetanus toxin mechanism of action in Torpedo electromotor system: a study on different steps in the intoxication process.

The mechanism of action of tetanus toxin was characterized in the electromotor system of Torpedo marmorata either at peripheral and central nervous system. The consecutive steps of the intoxication pathway were observed: (i) [125I]tetanus toxin specifically bound to neuronal plasma membranes isolated both from electric organ and electric lobe of Torpedo, exhibiting one and two binding sites respectively; (ii) [125I]tetanus toxin was internalized into nerve terminals and retrogradely transported to the electric lobe after its injection in the electric organ; (iii) finally, intracellular effect of tetanus toxin was studied either at electric organ and electric lobe membrane fractions. In both preparations tetanus toxin cleaved synaptobrevin, as detected by immunoblotting methods. In conclusion, our findings exhibit the presence of two different populations of acceptors for tetanus toxin in central and peripheral nervous system and show that synaptobrevin cleavage may account for intracellular toxicity in Torpedo.

Animals↗
PubMed1996

Differences in the multiple step process of inhibition of neurotransmitter release induced by tetanus toxin and botulinum neurotoxins type A and B at Aplysia synapses.

In order to gain insights into the steps (binding, uptake, intracellular effect) which differ in the inhibitory actions of tetanus toxin and botulinum neurotoxins types A or B, their temperature dependencies were investigated at identified cholinergic and non-cholinergic synapses in Aplysia. Upon lowering the temperature from 22 degrees C to 10 degrees C, extracellularly applied botulinum neurotoxin type A and B appeared unable to inhibit transmitter release whilst tetanus toxin exhibited a residual activity. Binding of each toxin to the neuronal membrane appeared virtually unaltered following this temperature change. By contrast, the intracellular effects of botulinum neurotoxin type B and tetanus toxin were strongly attenuated by temperature reduction whereas the inhibitory action of botulinum neurotoxin type A was only moderately reduced. Importantly, this discrepancy relates to the known proteolytic cleavage of different synaptic proteins by these two toxin groups. Since both the binding and intracellular activity of botulinum neurotoxin type A are minimally affected at 10 degrees C, its inability to inhibit neurotransmission at this low temperature when applied extracellularly indicated attenuation of its uptake. Due to the strict temperature dependence of the intracellular action of tetanus toxin and botulinum neurotoxin type B, but not A, an examination of the effects of changes in temperature on the internalization step was facilitated by the use of heterologous mixtures of the toxins' heavy and light chains. At 10 degrees C, heavy chain from tetanus toxin but not from botulinum neurotoxin type B mediated uptake of botulinum neurotoxin type A light chain. Collectively, these results provide evidence that, at least in Aplysia, the uptake mechanism for botulinum neurotoxin types A and B differs from that of tetanus toxin.

Animals↗
PubMed2005

Structural analysis of the catalytic domain of tetanus neurotoxin.

Clostridium neurotoxins, comprising the tetanus neurotoxin and the seven antigenically distinct botulinum neurotoxins (BoNT/A-G), are among the known most potent bacterial protein toxins to humans. Although they have similar function, sequences and three-dimensional structures, the substrate specificity and the selectivity of peptide bond cleavage are different and unique. Tetanus and botulinum type B neurotoxins enzymatically cleave the same substrate, vesicle-associated membrane protein, at the same peptide bond though the optimum length of substrate peptide required for cleavage by them is different. Here, we present the first experimentally determined three-dimensional structure of the catalytic domain of tetanus neurotoxin and analyze its active site. The structure provides insight into the active site of tetanus toxin's proteolytic activity and the importance of the nucleophilic water and the role of the zinc ion. The probable reason for different modes of binding of vesicle-associated membrane protein to botulinum neurotoxin type B and the tetanus toxin is discussed. The structure provides a basis for designing a novel recombinant vaccine or structure-based drugs for tetanus.

Amino Acid Sequence↗
PubMed1999

Tetanus toxoid and congenital abnormalities.

OBJECTIVE: To study the human teratogenic potential of tetanus vaccination during pregnancy. METHODS: Pair analysis of cases with congenital abnormalities and matched healthy controls was performed in the large population-based dataset of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1994. RESULTS: Of 35 727 pregnant women who had babies without any defects in the study period (control group), 33 (0.09%) were vaccinated with tetanus. Of 21563 pregnant women who had offspring with congenital abnormalities, 25 (0.12%) had tetanus vaccination. This difference was not significant (P = 0.39). The case-control pair analysis confirmed the safety of tetanus vaccination during pregnancy, particularly in the second and third months of gestation, i.e. during the critical period for congenital abnormalities. CONCLUSION: Tetanus vaccination during pregnancy appears not be teratogenic to the fetus. Thus, there is no contraindication, if the use of tetanus toxoid is necessary during pregnancy.

Abnormalities, Drug-Induced↗
PubMed1990

Tetanus immunization status and immunologic response to a booster in an emergency department geriatric population.

STUDY OBJECTIVES: Although effective procedures for the prevention of tetanus have long been available, serosurveys done since 1977 demonstrate that 49% to 66% of the elderly population lacks a protective antitoxin level (more than 0.01 IU/mL). This study was undertaken to assess the tetanus immunization status of patients presenting to an emergency department and to evaluate their immunologic response to a tetanus booster. SETTING: The study was conducted in a tertiary care ED. TYPE OF PARTICIPANTS: The patients enrolled were 65 or more years old and had breaks in their skin barriers. DESIGN: At each patient's initial presentation, pertinent demographic data and tetanus immunization history were recorded. The patient was then followed for 21 days. INTERVENTIONS: Each patient's antitoxin titer was determined on a serum sample by ELISA, and, if required by the Advisory Committee on Immunization Practices criteria, a booster was administered at the first visit. MEASUREMENTS AND MAIN RESULTS: Serum antitoxin assays were repeated on days 7, 14, and 21 after the initial visit until seroconversion (titer more than 0.01 IU/mL). Forty-four patients (55%) had protective levels at initial presentation, and in 36 (45%) the levels were not protective. Age and sex were not predictive of protection. Past military service and a definite history of three or more previous immunizations were good predictors of protection. Of 34 patients who were followed serially for inadequate initial titers, only 19 (56%) seroconverted by day 14. Patients who did not seroconvert were more likely to be older (P less than .05). CONCLUSIONS: This study demonstrated that a significant number of elderly patients lacked an initial protective level of tetanus antitoxin. Of these, 44% failed to seroconvert within 14 days and carried a potential risk of developing tetanus.

Aged↗
PubMed1998

Systemic and mucosal immune responses of mice to aluminium-adsorbed or aluminium-non-adsorbed tetanus toxoid administered intranasally with recombinant cholera toxin B subunit.

For the purpose of changing the immunization procedure of tetanus toxoid from intramuscular or subcutaneous injection, which has been in practice for a long time, to intranasal administration, we examined systemic and mucosal immune responses of mice to aluminium-adsorbed tetanus toxoid (aTT) and aluminium-non-adsorbed tetanus toxoid (nTT) inoculated intranasally with recombinant cholera toxin B subunit (rCTB). Intranasal immunization with aTT induced, at a concentration of 0.5 Lf, high levels of TT-specific serum IgG antibody titres and moderate levels of TT-specific serum IgA antibody titres in the presence and absence of rCTB. Induction of high or moderate levels of mucosal TT-specific IgA antibody responses was observed with and without rCTB in the lung, the nasal cavity, the small and large intestines and the vagina. Generally speaking, the co-administration of aTT and rCTB showed higher mucosal TT-specific IgA antibody titres when compared with the administration of aTT alone. In case of intranasal administration of nTT, the dose of 5 Lf was necessary and stimulated, only in the presence of rCTB (10 micrograms), high levels of tetanus toxoid (TT)-specific serum IgG antibody responses in all mice examined and moderate or slight levels of TT-specific IgA antibody responses in the nasal, pulmonary and small and large intestinal lavages of a few mice. All mice intranasally immunized with aTT alone or nTT and rCTB escaped onset of tetanus. This is the first report concerned with the mucosal adjuvant activity of an aluminium compound. Judging from these results, intranasal administration of aTT with and without rCTB or nTT with rCTB appears to be a very useful means for a vaccination against tetanus with respect to ease, safety, certainty, low cost and no need for an injection needle.

Administration, Intranasal↗
PubMed1999

[Trismus and tetanus vaccine].

BACKGROUND: Tetanus immunization, mandatory in France before the age of 18 months, is usually well tolerated. Fever and local reaction at the site of injection are the most common side effects. We report a case of trismus associated with tetanus immunization. CASE REPORT: A 12-year-old boy was hospitalized for a trimus which occurred 22 h after a booster dose of tetanus polio vaccine. The spasm of masticator muscles was so severe that the patient had difficulties in elocution and feeding. A moderate fever and a hypertonia of the lower limbs were associated. The trismus resolved itself within about 5 min after diazepam intravenous infusion. After examination of the buccal cavity, a local cause was discarded. The outcome was favorable and the clinical status of the boy was still normal six months later. DISCUSSION: Although no similar cases have been published, 13 other comparable adverse events associated with four vaccines containing tetanus toxoid were reported to the manufacturer within a period of 12 years. During the same period, more than 66 millions doses of vaccines containing tetanus toxoid were distributed. Trisms appeared to be more likely reported in adults after booster doses. Onset of the symptoms was between a few hours to 15 days. Outcome was always favorable. CONCLUSION: Although trimus associated to tetanus toxoid immunization is rare and remains unexplained, physicians should be aware of this adverse event.

Adult↗
PubMed2002

Identification of a binding site for ganglioside on the receptor binding domain of tetanus toxin.

The carboxyl-terminal region of the tetanus toxin heavy chain (H(C) fragment) binds to di- and trisialylgangliosides on neuronal cell membranes. To determine which amino acids in tetanus toxin are involved in ganglioside binding, homology modeling was performed using recently resolved X-ray crystallographic structures of the tetanus toxin H(C) fragment. On the basis of these analyses, two regions in tetanus toxin that are structurally homologous with the binding domains of other sialic acid and galactose-binding proteins were targeted for mutagenesis. Specific amino acids within these regions were altered using site-directed mutagenesis. The amino acid residue tryptophan 1288 was found to be critical for binding of the H(C) fragment to ganglioside GT1b. Docking of GD1b within this region of the toxin suggested that histidine 1270 and aspartate 1221 were within hydrogen bonding distance of the ganglioside. These two residues were mutagenized and found also to be important for the binding of the tetanus toxin H(C) fragment to ganglioside GT1b. In addition, the H(C) fragments mutagenized at these residues have reduced levels of binding to neurites of differentiated PC-12 cells. These studies indicate that the amino acids tryptophan 1288, histidine 1270, and aspartate 1221 are components of the GT1b binding site on the tetanus toxin H(C) fragment.

Animals↗
PubMed2002

An audit of the tetanus immunisation status of plastic surgery trauma and burns patients.

This study audits the tetanus immunisation management of plastic surgery trauma patients by their referring Accident and Emergency departments, and compares this to nationally published guidelines. We assessed 269 burns and trauma patients, referred from across South Wales, using a questionnaire together with their Accident and Emergency notes or letter. The precise question(s) that had been asked regarding the tetanus immunisation status of the patient, and the immunisation management based on the results of those questions, were recorded. The accurate tetanus immunisation status of the patient was established, the wound was assessed and further management was given as indicated. Only 16 patients had been asked whether they had received a course of tetanus, and 41 patients were not questioned about their tetanus immunisation status by the referring Accident and Emergency department. As a consequence of more accurate questioning, 73 patients (27%) required further action after their arrival in the Plastic Surgery unit. This audit has demonstrated that the management of tetanus immunoprophylaxis in plastic surgery trauma patients cannot be confidently left to the referring Accident and Emergency department but should form an integral part of the treatment at the admitting unit.

Adolescent↗
PubMed2005

[Assessment of the tetanus immune status in plasma donors of the Blood Donor Service of the Bavarian Red Cross].

BACKGROUND AND OBJECTIVE: Tetanus is nowadays largely under control, as a result of the preventive immunization programme. This prophylactic measure is important also because there is no specific treatment. In Germany tetanus occurs almost exclusively in elderly, not adequately immunized persons. This study determined the existing immune status of plasma donors in the region of Ingolstadt, Bavaria. SUBJECTS AND METHODS: Tetanus IgG antibody concentrations were measured with the ELISA test in serum of 1742 plasma donors (891 women, 851 men, age range 18-64 years). RESULTS: An adequately protective immunity against tetanus was found in 98.7% of men and 94.1% of women, while 5 women and 3 men had no protection. Demographic data indicated that the immunization protected males longer than females, regardless of age, especially in those over 50 years of age (85% of females, 99% of males). CONCLUSION: Adequate tetanus protection was achieved in the overwhelming majority of test subjects. This demonstrates that the prevalence of protection is lower in females than in males, a difference most marked in those over 50 years of age. It is important that this age group in particular should, as part of health care, be informed by their medical practitioner of the dangers of tetanus and the advantages of immunization.

Adolescent↗
PubMed2003

The genome sequence of Clostridium tetani, the causative agent of tetanus disease.

Tetanus disease is one of the most dramatic and globally prevalent diseases of humans and vertebrate animals, and has been reported for over 24 centuries. The manifestation of the disease, spastic paralysis, is caused by the second most poisonous substance known, the tetanus toxin, with a human lethal dose of approximately 1 ng/kg. Fortunately, this disease is successfully controlled through immunization with tetanus toxoid; nevertheless, according to the World Health Organization, an estimated 400,000 cases still occur each year, mainly of neonatal tetanus. The causative agent of tetanus disease is Clostridium tetani, an anaerobic spore-forming bacterium, whose natural habitat is soil, dust, and intestinal tracts of various animals. Here we report the complete genome sequence of toxigenic C. tetani E88, a variant of strain Massachusetts. The genome consists of a 2,799,250-bp chromosome encoding 2,372 ORFs. The tetanus toxin and a collagenase are encoded on a 74,082-bp plasmid, containing 61 ORFs. Additional virulence-related factors could be identified, such as an array of surface-layer and adhesion proteins (35 ORFs), some of them unique to C. tetani. Comparative genomics with the genomes of Clostridium perfringens, the causative agent of gas gangrene, and Clostridium acetobutylicum, a nonpathogenic solvent producer, revealed a remarkable capacity of C. tetani: The organism can rely on an extensive sodium ion bioenergetics. Additional candidate genes involved in the establishment and maintenance of a pathogenic lifestyle of C. tetani are presented.

Clostridium tetani↗
PubMed1979

Effect of tetanus toxin on the accumulation of the permeant lipophilic cation tetraphenylphosphonium by guinea pig brain synaptosomes.

Accumulation of the permeant lipophilic cation [(3)H]tetraphenylphosphonium (TPP(+)) by synaptosome preparations from guinea pig brain cerebral cortex is inhibited 1:10 by medium containing 193 mM K(+) and by veratridine. A further 1:10 to 1:15 decrease in TPP(+) uptake occurs under nitrogen and in the presence of mitochondrial inhibitors such as oligomycin, whereas starvation and succinate supplementation have no effect. These data indicate that, in analogy to intact neurons, there is an electrical potential (DeltaPsi, interior negative) of -60 to -80 mV across the synaptosomal membrane that is due primarily to a K(+) diffusion gradient (K(+) (in)-->K(+) (out)). The data also indicate that mitochondria entrapped within the synaptosome but not free mitochondria make a large contribution to the TPP(+) concentration gradients observed. Conditions are defined in which tetanus toxin binds specifically and immediately to synaptosomes in media used to measure TPP(+) uptake. Under these conditions tetanus toxin induces dose-dependent changes in TPP(+) uptake that are blocked by antitoxin and not mimicked by biologically inactivated toxin preparations. The effect of tetanus toxin on TPP(+) uptake is not evident in the presence of 193 mM K(+) or veratridine but remains under conditions known to abolish the mitochondrial DeltaPsi. Moreover, tetanus toxin has no effect on TPP(+) uptake by isolated synaptosomal mitochondria. The results thus define an in vitro action of tetanus toxin on the synaptosomal membrane that can be correlated with biological potency in vivo and is consistent with the in vivo effects of tetanus toxin on neuronal transmission.

Anaerobiosis↗
PubMed1999

Neuronal sensitivity to tetanus toxin requires gangliosides.

Tetanus toxin produces spastic paralysis in situ by blocking inhibitory neurotransmitter release in the spinal cord. Although di- and trisialogangliosides bind tetanus toxin, their role as productive toxin receptors remains unclear. We examined toxin binding and action in spinal cord cell cultures grown in the presence of fumonisin B(1), an inhibitor of ganglioside synthesis. Mouse spinal cord neurons grown for 3 weeks in culture in 20 microM fumonisin B(1) develop dendrites, axons, and synaptic terminals similar to untreated neurons, even though thin layer chromatography shows a greater than 90% inhibition of ganglioside synthesis. Absence of tetanus and cholera toxin binding by toxin-horseradish peroxidase conjugates or immunofluorescence further indicates loss of mono- and polysialogangliosides. In contrast to control cultures, tetanus toxin added to fumonisin B(1)-treated cultures does not block potassium-stimulated glycine release, inhibit activity-dependent uptake of FM1-43, or abolish immunoreactivity for vesicle-associated membrane protein, the toxin substrate. Supplementing fumonisin B(1)-treated cultures with mixed brain gangliosides completely restores the ability of tetanus toxin to bind to the neuronal surface and to block neurotransmitter release. These data demonstrate that fumonisin B(1) protects against toxin-induced synaptic blockade and that gangliosides are a necessary component of the receptor mechanism for tetanus toxin.

Animals↗
PubMed2002

Specific IgA subclass responses in serum and saliva: a 12-month follow-up study after parenteral booster immunization with tetanus toxoid.

Specific IgA subclass antibodies against tetanus toxoid in serum, parotid saliva, and whole saliva were quantified after booster immunization. Samples from 14 healthy individuals were collected before and 1, 6, and 12 months after subcutaneous injection with Duplex (0.23 ml tetanus toxoid 30 Lf/mL and diphtheria 7.3 Lf/mL). Samples of whole saliva were also collected after 2 weeks. Specific IgA1 and IgA2 subclass antibodies to tetanus toxoid were quantified by enzyme-linked immunosorbent assay (ELISA). In this quantitative method, chimeric IgA1 and IgA2 antibodies directed against NP (4-hydroxy-3-nitrophenacetyl) were used as standards. Total levels of IgA1 and IgA2 were measured using a nephalometer or ELISA. Immunization with tetanus toxoid resulted in raised mean values of specific IgA1 and IgA2 antibodies against tetanus toxoid in serum after 1 month. Compared with the baseline, the mean value of specific IgA1 antibodies showed a 2.6-fold increase (mean value 10.47 microgram/mL) in serum, and that of specific IgA2 antibodies a 2.7-fold increase (mean value 0.93 microgram/mL). Specific IgA subclass antibody levels in parotid and whole saliva were unchanged after 1 month. The ratio of specific IgA subclass antibodies to total IgA subclass antibodies was 3 to 10 times higher in parotid saliva compared with whole saliva. In conclusion, subcutaneous booster immunization with tetanus toxoid induced immune responses of both antigen-specific IgA1 and IgA2 subclass antibodies in serum with the same increase, whereas the levels of specific IgA subclass antibodies in secretory fluids were unchanged. The ratio of specific IgA subclass antibodies to immunoglobulins was higher in parotid saliva compared with whole saliva.

Adult↗
PubMed2006

Avidity of tetanus and Hib antibodies after childhood acute lymphoblastic leukaemia - implications for vaccination strategies.

AIM: To investigate the possible relationship between serum levels and avidities of antibodies against tetanus toxoid (TT) and Haemophilus influenzae type b (Hib) in children that were vaccinated after treatment for childhood acute lymphoblastic leukaemia (ALL). METHODS: The study groups were 31 paediatric patients with ALL and 18 healthy controls. All subjects were vaccinated with TT and a protein conjugated Hib vaccine. Antibody levels were analysed at three time points: At diagnosis of ALL, after cessation of treatment before vaccination and three weeks after vaccination. Avidity was measured twice, with a thiocyanate elution assay, at diagnosis of ALL and three weeks after vaccination. RESULTS: There was a correlation between level and avidity of tetanus antibodies after vaccination (r(s) = 0.59, P < 0.001). In the standard-risk and intermediate-risk ALL groups, all patients responded with protective levels of tetanus antibodies with normal avidity. In the high-risk ALL group 7/9 patients had subprotective levels of tetanus antibodies after vaccination and concomitantly the lowest avidity, implying poor protection against tetanus. No patients were found with low levels and low avidity of anti-Hib IgG, and 29/31 patients had full protection after a single dose of conjugated Hib-vaccine. CONCLUSION: The vaccination strategy after childhood ALL must be different for low-risk and high-risk ALL groups, since the high-risk group fail to elicit a recall response to tetanus.

Adolescent↗
PubMed1985

Fate of tetanus toxin bound to the surface of primary neurons in culture: evidence for rapid internalization.

We examined the nature of the tetanus toxin receptor in primary cultures of mouse spinal cord by ligand blotting techniques. Membrane components were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose sheets, which were overlaid with 125I-labeled tetanus toxin. The toxin bound only to material at or near the dye front, which was lost when the cells were delipidated before electrophoresis. Gangliosides purified from the lipid extract were separated by thin-layer chromatography and the chromatogram was overlaid with 125I-toxin. The toxin bound to gangliosides corresponding to GD1b and GT1b. Similar results were obtained with brain membranes; thus, gangliosides rather than glycoproteins appear to be the toxin receptors both in vivo and in neuronal cell cultures. To follow the fate of tetanus toxin bound to cultured neurons, we developed an assay to measure cell-surface and internalized toxin. Cells were incubated with tetanus toxin at 0 degree C, washed, and sequentially exposed to antitoxin and 125I-labeled protein A. Using this assay, we found that much of the toxin initially bound to cell surface disappeared rapidly when the temperature was raised to 37 degrees C but not when the cells were kept at 0 degree C. Some of the toxin was internalized and could only be detected by our treating the cells with Triton X-100 before adding anti-toxin. Experiments with 125I-tetanus toxin showed that a substantial amount of the toxin bound at 0 degree C dissociated into the medium upon warming of the cells. Using immunofluorescence, we confirmed that some of the bound toxin was internalized within 15 min and accumulated in discrete structures. These structures did not appear to be lysosomes, as the cell-associated toxin had a long half-life and 90% of the radioactivity released into the medium was precipitated by trichloroacetic acid. The rapid internalization of tetanus toxin into a subcellular compartment where it escapes degradation may be important for its mechanism of action.

Animals↗
PubMed1985

Inadequate tetanus protection among the rural elderly.

Three cases of tetanus occurring in a community in southern West Virginia in five years prompted a survey of the immunization status of the population of this region. An immunization history was obtained from 540 consecutive patients seen at three different health care facilities. Of these, 386 (71.5%) had received prior tetanus protection, 65 (12%) had never been immunized, and 89 (16.5%) were uncertain or had received incomplete immunization. Of the 65 nonimmunized patients, 54 (83%) were older than 50 and only 11 (17%) were younger. Compared to the total group sampled, significantly more nonimmunized patients (both male and female) were older than 50 (P less than .001). Also, significantly more of these never immunized individuals lived in rural areas (P less than .001). In a group of 222 patients identified as being at increased risk of having tetanus, 65 (29%) had never been immunized, 89 (40%) were of uncertain or incomplete status, and 68 (30.5%) had been immunized more than ten years previously. One hundred twenty (54%) high-risk patients were older than 50 and 103 (46%) were younger. Compared to the entire population sampled, high-risk patients included significantly more who were older than 50 (P less than .001). Also, significantly more high-risk patients lived in rural areas (P less than .007). When treating injured patients, it is important to recall that many older adults living in rural areas are not adequately immunized against tetanus. Such patients should receive human tetanus immunoglobulin, as well as tetanus toxoid.

Adolescent↗
PubMed1998

Prevention and control of tetanus in childhood.

Until recently, tetanus was estimated to be killing well over half a million children each year in the developing world, most commonly through neonatal tetanus, the incidence of which was around six per 1000 live births. Neonatal tetanus most commonly occurs through cord contamination of the umbilical stump. Vaccination of pregnant women and infants is an effective and inexpensive intervention that results in major health gains, although educating traditional birth attendants and mothers about safer birthing practices probably also lowers the incidence of tetanus. Recently, however, new vigor and strategies in immunization programs have resulted in substantial improvements. Immunization of pregnant women, and identification and targeting of areas and populations in which neonatal tetanus is occurring are especially effective. Tetanus will continue to be a pediatric problem until vaccination strategies are effectively implemented, surveillance systems are improved, and hygienic birthing practices become standard in all countries.

Child↗