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PubMed1979

Decrease of muscle extensibility and reduction of sarcomere number in soleus muscle following a local injection of tetanus toxin.

Slow soleus muscle in guinea pig developed within 4--6 days after local injection of a sublethal dose of tetanus toxin and 2--4 days after the first signs of local tetanus, a myostatic contracture characterized by a change in the passive tension--lengthening curve associated with a considerable decrease of the sarcomere number. It was demonstrated by clinical and EMG investigations that the soleus did have a tetanic spasm at least within the 2--4 day period of observation. When local tetanus was confined to slow soleus by functional suppression of rapid gastrocnemius and ankle flexor muscles, the decrease of the sarcomere number still persisted. This decrease failed to occur after section of the nerve supplying the soleus when associated with an injection of the tetanus toxin, and was much greater than when the soleus was passively shortened for the same period of time by plaster cast.

Animals↗
PubMed1981

Permanent tetraplegia as a consequence of tetanus neonatorum. Evidence for widespread lower motor neuron damage.

It is generally believed that no permanent neurological damage is found among survivors of tetanus neonatorum. Newborns dying shortly after the onset of tetanus also lack significant neurological abnormalities. In adults a variety of neuromuscular lesions have been reported; however, a uniform pathological picture is absent. We report a case of a newborn with severe tetanus in whom striking evidence of anterior horn neuronal damage was documented, causing permanent nonprogressive tetraplegia. We suggest that the mechanism responsible for this lesion involves the retrograde axoplasmic flow of tetanus toxin reaching the spinal cord via nerve endings in the infected umbilical cord stump.

Atrophy↗
PubMed1984

Cyclobenzaprine: effect on tonic vibration reflexes in local tetanus cat preparations.

Local tetanus was induced by the injection of toxin into the gastrocnemius-soleus muscle of cats. After 48 hr, longitudinal vibration of the muscle was used to elicit a reflex contraction (tonic vibration reflex, TVR). In other experiments, electrical stimulation of various regions of the central nervous system served to elicit contraction of the muscle alone or to facilitate a vibration (150 mu at 300 Hz)-induced tonic vibration reflex. In contrast with normal animals, tonic vibration reflex responses in local tetanus preparations of decerebrate cats were augmented after transection of the spinal cord at Cl. In decerebrate local tetanus preparations, a dose-related reduction of tonic vibration reflex responses, induced at all frequencies and amplitudes of muscle vibration, was observed after doses of 0.5 to 3.5 mg/kg (i.v.) of cyclobenzaprine. Muscle contractions induced by stimulation of the medial reticular formation and facilitation of tonic vibration reflex responses were more sensitive to the action of cyclobenzaprine than were similar responses activated through stimulation of Deiters' lateral vestibular nucleus. In spinal preparations, tonic vibration reflex responses were only moderately reduced after similar doses of cyclobenzaprine. Contractions induced by stimulation of the spinal cord (T6) and facilitated tonic vibration reflex responses were only moderately reduced, whereas the post-stimulus-induced facilitations were considerably attenuated. Thus, experiments in local tetanus preparations support further the concept that the major site of action of cyclobenzaprine is supraspinal, whereas its action upon spinal structures contributes to its overall skeletal muscle-relaxant activity.

Amitriptyline↗
PubMedDate not supplied

Cerebrospinal fluid changes in tetanus: raised proteins and immunoglobulins in patients with severe disease.

34 patients with tetanus were studied; all had normal serum proteins and albumin. Only 3 samples of cerebrospinal fluid (CSF) contained cells (a few polymorphonuclear leucocytes). Protein levels in the CSF were elevated in 26 patients (76.5%); 11 cases with clinically mild tetanus had a mean CSF protein of 400 +/- 250 mg/litre (normal 100-400 mg/litre), while the CSF protein levels of 10 patients who died (mean 1596 +/- 985 mg/litre; P less than 0.005) and those of 4 others with severe disease who absconded were much higher (mean 1220 +/- 562 mg/litre; P less than 0.005). As a group, 19 patients with severe disease including the 10 fatal cases also had significantly higher mean CSF protein values (1582 +/- 938 mg/litre; P less than 0.005) than did the mild cases. Immunochemical analysis of the proteins using a radial immunodiffusion assay showed that these proteins were immunoglobulin G (IgG). Polyacrylamide gel electrophoresis demonstrated an oligoclonal gammaglobulin pattern, suggesting intrathecal IgG synthesis in these patients. These studies suggest that measurement of CSF proteins may supplement clinical evaluation in tetanus and that there may be a local immune stimulus (perhaps a tetanus antigen or to some other mitogen) in the central nervous system.

Adolescent↗
PubMed1984

Affinity chromatographic purification and characterization of two iodinated tetanus toxin fractions exhibiting different binding properties.

Highly purified iodinated tetanus toxin preparations separate on ganglioside affinity columns into two distinct (A and B) fractions representing about 20% and 75% of the iodinated toxin, respectively. Fraction A, eluted by 1% NaCl, migrates like native tetanus toxin (150,000 mol. wt) on SDS polyacrylamide gel electrophoresis. It forms an aggregate of molecular weight approximately 360,000 on Sephacryl S-300 gel permeation chromatography in the presence of detergent and contains two isoforms on preparative chromatofocusing. Fraction A binds poorly to neurons in tissue culture or to synaptosomal membrane preparations. It retains, however, its antigenicity and biotoxicity. Fraction B, eluted by 6% NaCl, binds effectively to gangliosides and also to neurons or synaptosome preparations. It has a similar molecular weight and chain composition to the native toxin and displays two isoforms, precipitable during chromatofocusing. Fraction B possesses similar binding, immunological and toxic properties to the original iodinated tetanus toxin. Following excessive iodination (4-6 mCi/mg protein), toxicity can be remarkably reduced. Unlabeled toxin shows a similar chromatographic pattern to the iodinated toxin on affinity columns, suggesting that a large portion (30% by protein and 55% by toxicity) of the toxin has a poor affinity for gangliosides. The molecular pharmacokinetics of tetanus toxin with respect to affinity toward ganglioside-dependent and ganglioside-independent receptors needs re-evaluation.

Animals↗
PubMed1986

Botulinum A neurotoxin unlike tetanus toxin acts via a neuraminidase sensitive structure.

The binding and effects of tetanus and botulinum A neurotoxins were studied on mouse spinal cord cultures treated with neuraminidase. In untreated cultures both neurotoxins blocked synaptic transmission. Treatment of the cell cultures with neuraminidase, 25 mU/ml for 24 hr, decreased the potency of botulinum A neurotoxin. At 7 X 10(-11) M no toxin effect on inhibitory or excitatory synapses was observed, whereas at higher concentrations of the toxin the concentration-response curve was shifted to the right by a factor of about 30. Surprisingly, the action of tetanus toxin over a large concentration range was unaffected by pretreatment of the neurones with the enzyme. Accordingly, neurones treated with neuraminidase failed to bind 125I-botulinum A neurotoxin, whereas labelled tetanus toxin was still fixed by cell bodies, as well as by neurites, as shown by histoautoradiography. Chromatographic extraction of gangliosides from cultures prelabelled with 14C-glucosamine showed a dramatic loss in the contents of polysialogangliosides following treatment with neuraminidase. Our results indicate that neuraminidase-sensitive structures might be important for the action of botulinum A neurotoxin. The effect of tetanus toxin appears to be mediated by a different site which is insensitive to neuraminidase.

Action Potentials↗
PubMed1988

Tetanus and botulinum toxins block the release of acetylcholine from slices of rat striatum and from the isolated electric organ of Torpedo at different concentrations.

Tetanus toxin, like botulinum toxin type A, blocks cholinergic synaptic transmission at the central and peripheral nervous systems. Nevertheless, the diseases induced by the two toxins are different since tetanus toxin induces a spastic paralysis and botulinum toxin elicits a flaccid paralysis. Thus, we have investigated the sensitivity of a central and a peripheral cholinergic synapse to these two toxins. We have studied the action of both poison on the release of acetylcholine from slices of the rat striatum and from the isolated electric organ of Torpedo, which is homologous to the neuromuscular junction. Acetylcholine release from the rat striatum was continuously monitored by a chemiluminescent method. The secretion of acetylcholine from the electric organ was estimated both by measuring the amplitude of the evoked electrical discharge from stacks of electroplaques, and by continuously monitoring the neurotransmitter release from isolated nerve terminals. Tetanus toxin blocks the electrical discharge of electric organ prisms, and also impairs the release of acetylcholine from the Torpedo electric organ nerve endings. Our results on acetylcholine release show that tetanus toxin is more potent than botulinum toxin type A at the central cholinergic synapse (tetanus/botulinum toxins potency ratio about 100-200) whereas botulinum toxin is the most potent at the peripheral cholinergic synapse (botulinum/tetanus toxins potency ratio about 100).

Acetylcholine↗
PubMed1989

Gangliosides mediate inhibitory effects of tetanus and botulinum A neurotoxins on exocytosis in chromaffin cells.

Bovine chromaffin cells in monolayer culture were preloaded with 3H-NA (noradrenaline) and subsequently stimulated with carbachol. Botulinum A neurotoxin partially inhibited the evoked release of 3H-NA and the basal efflux of the hormone. The inhibition of evoked release did not exceed 40%, although the cells were exposed to 10 micrograms/ml of toxin for 6 days. The inhibitory effect of botulinum A neurotoxin was neutralized by its antibodies. In contrast to botulinum A neurotoxin, tetanus toxin at even higher concentrations did not influence evoked release. This difference in sensitivity could be explained by the ganglioside pattern of chromaffin cells. Ganglioside GD1a, a putative receptor for botulinum A neurotoxin, could be identified in lipophilic extracts, whereas the tetanus toxin binding gangliosides GT1b and GD1b could not be detected by means of thin-layer chromatography. Treatment of the cells with neuraminidase abolished both, GD1a and the inhibitory effect of botulinum A neurotoxin. Incubation of chromaffin cells with a mixture of gangliosides (21% GM1, 44% GD1a, 15% GD1b, 20% GT1b) not only increased the efficacy of botulinum A neurotoxin but also made the cells sensitive towards tetanus toxin. The concentration-response curve of botulinum A neurotoxin was shifted to the left about five-fold and the maximum inhibition of evoked release was increased up to 60%, even though the cells were exposed to the toxin for 3 days only. In contrast, the maximum inhibition that could be achieved by tetanus toxin was 40%. The results indicate that polysialogangliosides are important for the intracellular accumulation of these clostridial neurotoxins.

Adrenal Medulla↗
PubMed1990

Tetanus toxin blocks potassium-induced transmitter release and rearrangement of intramembrane particles at pure cholinergic synaptosomes.

We have studied the action of tetanus toxin on the release of acetylcholine from a subcellular fraction of cholinergic nerve terminals (synaptosomes) isolated from the Torpedo electric organ. We have also studied the morphological changes induced by chemical stimulation on the presynaptic plasma membrane of poisoned synaptosomes. These changes were studied by means of freeze-fracture techniques. We found that tetanus toxin blocks the release of acetylcholine from isolated nerve terminals in a dose-dependent manner. The maximal inhibition is achieved at a concentration of 12.5 nM in 10 min. This effect is prevented by tetanus toxin antiserum. Tetanus toxin also blocks the rearrangement of intramembrane particles at plasma membrane of poisoned synaptosomes, specifically the decrease of small (less than or equal to 9.5 nm diameter) intramembranous particles at the protoplasmic hemimembrane leaflet and the increase of large (greater than 9.5 nm diameter) intramembrane particles at the external hemimembrane leaflet induced by potassium stimulation. These results suggest that intramembrane particle rearrangement could be related to acetylcholine secretion.

Acetylcholine↗
PubMed1985

Reduced antigen-induced proliferation and surface Ia expression of peripheral blood T cells following tetanus booster immunization.

Immune regulatory changes after tetanus booster immunization were studied by measuring T-cell proliferation and surface Ia expression after cocultivation of T cells with autologous tetanus toxoid pulsed macrophages. Both soluble and aluminium-absorbed tetanus toxoid was used as antigen. T cells proliferated readily prior to immunization, if TTAl was used as an antigen (when proliferation was measured by the uptake of [3H]thymidine: mean +/- SEM 64.5 +/- 11.5 X 10(3) dpm). The proliferative response of the peripheral blood T cells studied decreased significantly to 29.3 +/- 9.3 X 10(3) dpm (P less than 0.01) 1 week postimmunization, returned to preimmunization values after 2 to 4 weeks, and increased further thereafter. Surface Ia expression paralleled the proliferative response at every point pre- and postimmunization. Serum antibody levels increased steadily during the first weeks. These results demonstrate a functional parallel to the previously described changes in T-cell phenotypes (M. M. Eibl, J. W. Mannhalter, and G. Zlabinger, N. Engl. J. Med 310, 198, 1984) after tetanus booster immunization.

Adult↗
PubMed1987

Quantitative estimation of diphtheria and tetanus toxoids. 2. Single radial immuno-diffusion tests (Mancini) and rocket immuno-electrophoresis test in comparison with the flocculation test.

The concentration in Lf units, of an unknown diphtheria or tetanus toxoid preparation is estimated in the flocculation test relative to reference preparations of tetanus and diphtheria antitoxins, respectively. By replacing the antitoxin reference preparations with toxoid reference preparations it should be possible to use immunological methods other than the flocculation test for the quantitative estimation of toxoids in Lf units. A number of diphtheria and tetanus toxoids were tested by rocket immuno-electrophoresis and single radial immuno-diffusion (Mancini test). The concentrations of the unknown toxoids were expressed relative to a diphtheria toxoid calibrated in Lf units (DIFT) and a tetanus toxoid calibrated in Lf units (TEFT), respectively. These two toxoid preparations are regarded as candidates for establishment as international standard preparations. The results obtained in the two tests were compared with those obtained in the flocculation test. In most cases the differences between the results did not exceed 10%. It is concluded, therefore, that the rocket electrophoresis or the radial immuno-diffusion tests can be used as alternatives to the flocculation test.

Diphtheria Toxoid↗
PubMed1994

Synthetic peptide antigens of tetanus toxin.

In this study the immunochemical structure of the heavy chain polypeptide from tetanus toxin was studied. Numerous antigenic determinants were identified by probing a set of overlapping peptides derived from the amino acid sequence of tetanus toxin with polyclonal anti-toxoid antibody preparations. Synthetic antigens representing continuous epitopes were prepared and used to immunize mice. The capacity of the resulting anti-peptide antibodies to react with tetanus toxin in vitro and in vivo was determined. The majority of antibodies bound to tetanus toxin and three epitopes capable of eliciting neutralizing antibodies were identified.

Amino Acid Sequence↗
PubMed1981

Expression of Thy-1, H-2, and NS-4 cell surface antigens and tetanus toxin receptors in early postnatal and adult mouse cerebellum.

The expression of several cell surface components (Thy-1, H-2 and NS-4 antigens and tetanus toxin receptors) was studied by indirect immunofluorescence in situ using histological sections and in vitro using freshly dissociated and cultured cells from mouse cerebellum. Thy-1 alloantigen is expressed in adult cerebellum predominantly in neuron-rich regions, i.e. molecular, Purkinje cell, and granular layers, however, it is not detectable at postnatal day 8. In cerebellar cultures of 6-day-old mice Thy-1 is absent from more than 99% of all cells when these are maintained as monolayers in vitro for up to 3 days. After 4 days in vitro some GFA protein-positive astrocytes and some fibronectin-positive fibroblast-like cells start to express Thy-1 antigen. After 14 days in vitro not all fibroblast-like cells and astrocytes are Thy-1 antigen-positive. Neurons with small cell bodies and oligodendrocytes never express Thy-1 at any stage examined. H-2 is not expressed sufficiently to be detectable in histological sections in early postnatal or adult cerebellum. In cerebellar cultures of 6-day-old mice H-2 becomes detectable on some fibroblast-like cells and some astrocytes after 7 days in culture. In histological sections of adult and early postnatal cerebellum NS-4 antigen and tetanus toxin receptors are expressed at higher levels on more mature granule cells. In cerebellar cultures NS-4 antigen and tetanus toxin receptors are expressed on neurons. Occasionally some astroglia can also show detectable levels of expression. NS-4 antigen is also present on some 04 antigen-positive oligodendrocytes, while tetanus toxin receptors are never detectable on these cells.

Age Factors↗
PubMed1989

Immunoenhancement with combined rabies and aluminium-adjuvanted tetanus vaccines.

The effect of combining tetanus toxoid in the same syringe with purified Vero cell rabies vaccine (PVRV) was investigated in the 2-1-1 regimen of PVRV. The 2-1-1 regimen alone was as immunogenic as the five-dose regimen, while saving one dose of vaccine and two clinic visits. When aluminium hydroxide-adsorbed tetanus toxoid was used to dissolve PVRV on days 0 (one of the two doses) and 21, the anti-rabies antibody was significantly increased. Aluminium-free tetanus toxoid was ineffective, suggesting that immunoenhancement was due to aluminium adjuvant. In addition, anti-tetanus antibody was unaffected and the side-effects were not increased by such mixing.

Adjuvants, Immunologic↗
PubMed1994

Biodegradable microspheres as controlled-release tetanus toxoid delivery systems.

Purified tetanus toxoid, a high-molecular-weight protein, was entrapped within poly(L-lactic acid) (PLA) and poly(D,L-lactic/glycolic acid) (PLGA) microspheres prepared by either a solvent extraction or a solvent evaporation method carried out in a multiple emulsion system (water-in-oil-in-water). The physical integrity and antigenicity of the protein treated under different processing conditions were investigated. A reduction of antigenicity that was related to the percentage of aggregated protein was noticed under some experimental conditions. This partial loss of antigenicity was associated with the lyophilization process and affected by the nature of the organic solvent. All types of microspheres prepared with different molecular weight PLA and PLGA displayed a high protein-loading efficiency (> 80%) but their size was strongly influenced by polymer molecular weight (3000 versus 100,000). Protein release pattern was influenced by both polymer molecular weight and composition (PLA versus PLGA). A constant release pattern after an induction period of 10 days was observed for microspheres composed of high-molecular-weight polymers (PLA and PLGA). The release rate was lower from PLA microspheres than from PLGA microspheres. In contrast, a continuously increasing release rate preceded by a burst was observed for low-molecular-weight (3000) PLGA microspheres. Microencapsulated tetanus toxoid was significantly more immunogenic in mice than fluid toxoid as determined by IgG anti-tetanus antibody levels and neutralizing antibodies. However, the magnitude and duration of the antibody response did not differ significantly from a similar dose of aluminium phosphate-adsorbed toxoid. We conclude that microencapsulated tetanus toxoid shows significant adjuvant activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Interaction of Haemophilus influenzae type b conjugate vaccines with diphtheria-tetanus-pertussis vaccine in control tests.

The effects of combining three Haemophilus influenzae type b (Hib) capsular polysaccharide vaccines, conjugated to different proteins, with DTP vaccine on the subsequent control testing were examined. The addition of the Hib vaccines had little effect on the reactogenicity or the potency of the whole-cell pertussis component. The potency of, and antibody responses to, the diphtheria component were also unaffected in all three combinations. However, combination with the Hib vaccine comprising polysaccharide conjugated to tetanus toxoid resulted in a fivefold potentiation of the tetanus potency and large increases in the antibody responses to tetanus toxin and toxoid and Hib polysaccharide. These results have implications for the control testing of combined vaccines containing a whole-cell pertussis component and Hib polysaccharide-tetanus protein conjugate vaccine.

Animals↗
PubMed1981

Tetanus toxin: a marker of amphibian neuronal differentiation in vitro.

Tetanus toxin may be used to specifically mark neurones in cultures of mammalian and chick nervous tissue. We have prepared cultures of neural ectoderm and underlying mesoderm from late neurula Xenopus laevis embryos, and shown that in this Amphibian species, neurones are labelled by tetanus toxin. Aggregate cultures of ectoderm and mesoderm from late blastula embryos were than examined to determine whether tetanus toxin might be used to mark neuronal precursor cells. Our observations suggest that this is not possible; neurones are seen to have acquired tetanus toxin receptors only after they have become morphologically differentiated.

Animals↗
PubMed1991

Cooperative action of the light chain of tetanus toxin and the heavy chain of botulinum toxin type A on the transmitter release of mammalian motor endplates.

Purified heavy chain of botulinum toxin type A and light chain of tetanus toxin were combined to form a chimeric toxin. It was active on the mouse phrenic nerve-hemidiaphragm with a potency 6 times higher than that of native tetanus toxin. Electrophysiological data from poisoned neuromuscular junctions revealed that the pattern of nerve-evoked and spontaneous transmitter release was equivalent to that seen with tetanus toxin i.e. asynchronous release, and did not resemble that after botulinum toxin type A poisoning. We conclude that the light chain of tetanus toxin alone is responsible for the characteristic effects on spontaneous and nerve-evoked transmitter release of the native toxin and that these properties can be introduced into a new, more potent complex with the heavy chain of botulinum toxin A.

Animals↗