Competition between tetanus toxoid and toxin

Experiments on albino mice showed that preliminary injection of tetanus toxoid increases the resistance of animals to tetanus toxin, as manifested by an increase in LD₅₀. The effect is enhanced by increasing the dose of toxoid or by giving it in fractional doses. The use of protagon and unpurified mitochondrial fraction, isolated from the brain, as receptor of tetanus toxin in the nerve tissue revealed competition for substrate between the tetanus toxoid and toxin. The results of these experiments confirm the writers' earlier hypothesis that the tetanus toxin molecule contains different functional groups responsible for binding the toxin with the receptor in brain tissue, for the pathogenic action of the toxin, and for the binding of the toxin with antitoxin.     

The mechanism of immunity to tetanus toxin at early periods after active immunization

Summary Experiments on white rats have established that after primary immunization with a single, relatively large dose of adsorbed tetanus toxoid the resistance of the animals to tetanus toxin develops prior to the appearance of antitoxin in the blood. However, the administration at that period of various substances, such as tetanus toxin andBac. perfringens toxoid, or cortisone, makes the animals react by rapid production of antitoxin in the blood in detectable amounts. This phenomenon is regarded as a nonspecific stimulation of the antitoxin-forming apparatus (regional lymph modes) periods after immunization and in the absence of antitoxin in the blood at the moment of toxin administration.Presented by Active Member AMN SSSR V. N. Chernigovskii     

Routes of tetanus toxin entrance into the central nervous system and some problems of experimental tetanus pathogenesis

Summary As shown by experiments on monkeys (table 1) and dogs (table 2), the main and direct route of passage of the tetanus toxin from the muscles into the spinal cord are the anterior spinal roots. This result coincides completely with the data of previous investigations on albino rats, guinea pigs, rabbits and cats [4,5,6]. After blocking the circulatory route of spread of the toxin by means of antiserum, the administration of toxin into the muscles of the posterior extremity causes a fatal ascending tetanus in dogs (Fig.2); by the time of the animal's death the antitoxin may circulate in the animal's blood (table 3). Thus, the spread of the toxin with the blood is not a required condition for the development of general fatal tetanus. However, this factor plays an important role in determining the clinical form and the outcome of the disease. Both in the monkeys (Fig.1, d, f) and in dogs (Fig. 2, f, e) there is present a phenomenon of generalization of excitation in the central nervous system upon stimulation of the extremity into which the toxin has been introduced; this phenomenon is characteristic of the ascending general tetanus and was described by the authors earlier (3). A discussion is given on the clinical forms of the disease and the route of the tetanus spread of toxin in connection with the current concepts on the pathogenesis of tetanus.(Presented by AMN SSSR Active Member of V. V. Parin) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 52, No. 11, pp. 35–43, November, 1961     

Relation between the level of specific antitoxin in the blood and the degree of immunity to tetanus toxin in wilte mice immunized with adsorbed tetanus toxoid

Summary Levels of the specific antitoxin (AE) in blood and the immunity intensity against tetanus toxin (LD₅₀) were simultaneously studied on white mice immunized with the adsorbed tetanus toxic. The blood for the antitoxin determination was taken from the blood vessels of the tail (cut off): AE and LD₅₀ increased with an increase of the antigen dose, the growth rate of the latter being much greater. The ratio between the growth rates of AE and LD₅₀ changed in the course of the development of immunity.LD₅₀ being constant, AE level in the blood corresponded to the period elapsed after the antigen injection.Presented by Member of the Academy of Medical Sciences of the USSR, P. F. Zdrodovsky     

Routes of entrance of tetanus toxin into the central nervous system and certain questions in the pathogenesis of experimental tetanus

Summary In donkeys as well as in other animals tetanus toxin injected into the muscles of the shank passes along the sciatic nerve and reaches the spinal cord through the anterior roots (Table 1). In the usual conditions, after the administration of the toxin (lethal doses) into the muscles of extremities the course of tetanus in these animals is of the so-called descending type (Fig. 1). After blocking the circulatory route of the toxin spread by means of tetanus antiserum, tetanus develops according to the ascending type (Fig. 2), starting with local tetanus with increased electrical activity of the muscles into which the toxin was injected (Fig. 3c). Local tetanus may also be induced by minimal doses of the toxin (1/50 DLM) administered into the muscles of extremities (Fig. 3b). Ascending tetanus is based upon toxin spread from the site of administration along the regional nerve and the anterior roots into the spinal cord, whereas descending tetanus is based upon toxin spread by way of circulation.When the main mass of toxin enters the circulation and a relatively small amount of it passes along the nerve general tetanus develops before local tetanus has time to appear. This mechanism takes place in donkeys. It is suggested that the mechanism of descending tetanus in other cases, including man, is analogous to this.(Presented by Active Member of the Akad. Med. Nauk SSSR, V. V. Parin) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 52, No. 12, pp. 30–38, December, 1961     

Ultrastructural changes in experimental tetanus

In experiments on rats an electron-microscopic study was undertaken of the caudal part of the hypothalamic-hypophyseal neurosecretory system in experimental tetanus. Activation of the system was found in the early stages after injection of tetanus toxin, evidently on account of the injection of heterologous protein, for a similar effect was given by injection of the inactivated toxin. Parallel changes were observed in vascular permeability, lipid metabolism, and the clotting system of the blood.Central Research Laboratory, Rostov Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR A. M. Chernukh.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 85, No. 4, pp. 489–493, April, 1978.     

The incubation period in passive immunity to tetanus toxin

Summary It was established in experiments on white rats that the time of the maximal immunity against tetanus depends on the method of introduction of antitetanus serum and tetanus toxin. The resistance of the animals to the toxin depends directly on the content of the antitoxin in their blood. The mechanism of action of tetanus antitoxin and conditions of appearance of the passive immunity are discussed.Presented by Active Member of the Acad. Med. Sci. USSR V. N. Chernigovskii     

Some properties of low-molecular-weight tetanus antitoxin

After intravenous injection of tetanus antitoxin obtained by tryptic digestion of Diaferm-3 horse immunoglobulin, purification, and concentration of the active fragments, the antitoxin was excreted by rabbits 3 times faster than after injection of the original Diaferm-3 antitoxin. After injection of the fragmented antitoxin its excretion continued until the 6th days, whereas after injection of Diaferm-3 antitoxin its excretion continued until the 19th day; in the first case much less antitoxin was excreted than in the second (2% and 3.5% respectively). In both cases the antitoxin excreted in the urine consisted of monovalent Fab' fragments which caused delay of precipitation in the cross reaction in agar gel between tetanus toxoid and antitetanus serum. The Fab' gragment obtained by this method possessed anaphylactogenic properties.L'vov Medical Institute. Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 81, No. 3, pp. 331–333, March, 1976.     

Determination of cerebrosides in different parts of the brain of rabbits suffering from generalized tetanus

Summary The cerebroside content of the subcortical formations of the cerebral hemispheres of rabbits falls following injection of a lethal dose of tetanus toxin into a hind leg to a minimum on the 7–8th day, when the animals are moribund. This effect is more pronounced in the hemisphere contralateral to the injection site.(Presented by S. E. Severin, Active Member, Academy of Medical Sciences, USSR) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 51, No. 4, pp. 66–70, April, 1961     

Use of acetylcholinesterase reactivators and of central cholinolytics in the treatment of experimental tetanus

The therapeutic efficacy of acetylcholinesterase reactivators dipyroxime and isonitrozin, and also of the central cholinolytics benactyzine and adiphenine, was studied in experiments on rabbits with tetanus produced by intravenous injection of one lethal dose of tetanus toxin. Dipyroxime in a dose of 25 mg/kg had no therapeutic action, and in doses of up to 30–40 mg/kg caused death of the animals. Benactyzine and adiphenine, in a dose of 3–4 mg/kg, abolished tonic convulsions for 1.5–2 h, and isonitrozin in a dose of 25 mg/kg did so for 4–5 h. After combined administration of reactivators and cholinolytics, convulsions were abolished for 4–5 h.Central Research Laboratory, L'vov Medical Institute. (Presented by Academician of the Academy of Medical Sciences of the USSR A. M. Chernukh.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 7, pp. 40–42, July, 1978.     

Neuropathological effects of injection of tetanus toxin into certain structures of the rat brain

The neuropathological effects of local injection of tetanus toxin (TT) into various structures of the brain were studied in experiments on rats. Definite neuropathological changes were observed in the animals, different from those found after injection of TT elsewhere. As a rule the action of TT in a given region of the brain was local. The experiments confirm the theory of generator mechanisms of neuropathological syndromes, according to which specific manifestations of the corresponding syndrome are determined by the location of a generator of pathologically enhanced excitation in a certain brain structure.Laboratory of General Pathology of the Nervous System, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. M. Chernukh.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 2, pp. 124–126, February, 1979.     

Effect on Diphtheria Immunity of Combined Tetanus and Diphtheria Booster Vaccination in Adults

 The effect of a single booster injection of an adult formulation of a combined diphtheria–tetanus vaccine (Td) on diphtheria-specific immunity was evaluated. The booster injection, containing 2 IU diphtheria toxoid per dose, was given as part of the surgical wound management for adults with open soft tissue injuries. Diphtheria antitoxin concentrations were determined in serum samples from 534 patients (199 women and 335 men, aged 18–70 years) using an enzyme immunoassay and a tissue culture toxin neutralization assay. Seroimmunity against diphtheria toxin was classified at three levels: susceptibility, basic protection, and full protection against the toxic manifestations of the disease. Before vaccination, 27.1% of the subjects were susceptible to diphtheria, 26.5% had basic protection, and 46.4% were fully protected. Six weeks (minimum 25 days, maximum 98 days) after a single booster injection, 89.7% of the subjects achieved full protection against diphtheria, and only 3.9% had antitoxin levels below the protective level. The median increase from the prevaccination to postvaccination antitoxin concentration was found to be 14-fold (4.4–47; quartiles Q25 to Q75). The change in antitoxin levels after revaccination was higher in older age groups (P<0.001), whereas neither sex (P=0.86) nor the country of previous immunization with a different national immunization schedule (P=0.61) had a significant influence on the revaccination effect. Systemic adverse reactions were rare, and local reactions of clinical significance were reported in only 1.9% of subjects.     

Antibodies against the light chain of tetanus toxin in human sera.

Tetanus toxoid elicits protective antibodies against tetanus toxin in humans and animals. It has been reported that antitoxin from immunized humans contains no anti-light chain antibodies, based on immunodiffusion and quantitative precipitin analyses. We confirmed the absence of precipitating anti-light chain antibodies in tetanus immune globulin. However, the presence of antibodies against the light chain of the toxin was shown by direct binding and inhibition analyses, using enzyme-linked immunosorbent assays. Using a neutralization inhibition test, we also found that about one-fourth of the neutralizing antibodies in tetanus immune globulin are directed against the light chain. These results suggest that the light chain of tetanus toxin contains immunogenic determinants and that antibodies directed against it may have a role in the prevention of tetanus or treatment of tetanus or both.     

Experimental simulation of neuropathological syndromes by creating pathologically enhanced excitation generators in the rabbit hypothalamus

Experiments on rabbits under free behavior conditions showed that injection of tetanus toxin into motivation-emotiogenic pacemaker regions of the hypothalamus, with the object of producing generators of pathologically enhanced excitation in those regions gives rise to stable and prolonged disturbances of the animals' motivational-emotional behavior. These changes were expressed as a marked increase in foodgetting activity by the animals, including an increase in secondary motivational responses, increased motor activity, the appearance of an excessive number of automated chewing movements, aggressiveness, a fear reaction, and the corresponding autonomic changes. The character of these responses depended on the region into which the toxin was injected and on its dose. Investigations showed that the formation of long-acting generators of pathologically enhanced excitation in motivation-emotiogenic pacemaker centers of the hypothalamus with the aid of tetanus toxin can be used to simulate psychopathological states in animals. The results confirm, by the use of this new model, the theory of generator mechanisms of neuropathological syndromes characterized by hyperactivity of systems.Laboratory of General Pathology of the Nervous System, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR. Laboratory of Emotions and Emotional Stresses, P. K. Anokhin Institute of Normal Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 84, No. 10, pp. 405–408, October, 1977.     

Tetanus toxin and synaptic inhibition in the substantia nigra and striatum of the rat.

1. The effects of tetanus toxin were determined on GABA-mediated synaptic inhibition of substantia nigra neurones evoked by striatal stimulation and on the presumed dopamine- and 5-hydroxytryptamine-mediated synaptic inhibition of striatal neurones evoked by nigral and dorsal raphe nucleus stimulation, respectively, in the urethane-anaesthetized rat. 2. Following an intranigral injection of tetanus toxin, striatal-evoked inhibition of substantia nigra neurones, which is sensitive to bicuculline, was rapidly abolished. This effect was not accompanied by any significant change in the responses of nigral neurones to ionophoretically administered GABA or other putative neurotransmitters and thus indicates a presynaptic site of action of the toxin. 3. The rate of onset of action of the toxin in the substantia nigra was extremely rapid (1-4 min) and appeared to be related to the rate of activation of the inhibitory pathway. 4. Injections into the substantia nigra of tetanus toxin neutralized with antitoxin had no significant effect on striatal-evoked inhibition in the substantia nigra. 5. Injections of tetanus toxin into the striatum failed to influence the inhibition of striatal neurones evoked by stimulation of the ipsilateral substantia nigra or the dorsal raphe nucleus, suggesting that tetanus toxin does not impair monoamine-mediated inhibition in the central nervous system. 6. Synaptic excitation which preceded substantia-nigra-evoked inhibition in striatal neurones and which occasionally preceded striatal-evoked inhibition in nigral neurones was also unaffected by tetanus toxin. 7. It is suggested that tetanus toxin selectively abolishes GABA-mediated synaptic inhibition in the central nervous system and may be a useful tool in the identification of such synaptic inhibitory mechanisms.     

Analgesic effects of inhibitory mediators in a pain syndrome of spinal origin

A pain syndrome of spinal origin was induced by creating a generator of pathologically enhanced excitation (GPEE) in the dorsal horn of the lumbosacral segments of the spinal cord in rats with the aid of tetanus toxin, strychnine, penicillin, KCl, and ouabain. The substances were applied to the dorsal surface of the spinal cord. An agar wafer method was developed to enable local prolonged and dose-dependent effects of the substances applied to be produced and studied in freely behaving animals. The effects of the inhibitory mediators were determined by the nature of the generator produced and, in particular, by the state of the mediator-receptor system of the neuronal membranes of cells forming in GPEE. Glycine prevented the development of the pain syndrome in the case of generators induced with tetanus toxin, penicillin, KCl, and ouabain. GABA was effective in the case of generators produced by tetanus toxin, potassium, and ouabain. Injection of glycine into the region of a generator produced by tetanus toxin at the height of development of the pain syndrome abolished that syndrome for the duration of action of the mediator. The pain syndrome could thus be specifically abolished or prevented for the period of action of the mediator by depressing the GPEE with the appropriate inhibitory mediators, depending on the nature of the GPEE, i.e., on the features distinguishing the state of the mediator apparatus of its neurons. Under these conditions inhibitory mediators, although not analgesics in the pharmacological sense, have a specific analgesic effect.Laboratory of General Pathology of the Nervous System, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 87, No. 6, pp. 525–528, June, 1979.     

Induction of a Mucosal Antitoxin Response and Its Role in Immunity to Experimental Canine Cholera

The induction of a jejunal antitoxin response was studied in dogs immunized with cholera toxin or toxoid. Single doses of toxoid given subcutaneously (s.c.) or of toxin given intraluminally (i.l.) were each effective in priming the mucosal immune system, whereas toxoid given i.l. was much less effective. In contrast, toxin and toxoid given i.l. were each effective as booster antigens. The local secondary response was rapid and brief, the peak occurring at about 7 days after i.l. boosting and declining by 90% after 2 more weeks. After s.c. priming and i.l. boosting with toxoid, antitoxin-containing plasma cells appeared predominantly in the portion of jejunum exposed to the i.l. booster. The appearance of antitoxin-containing plasma cells in jejunal lamina propria correlated with the amount of antitoxin recovered in jejunal washings which, in turn, correlated with protection against challenge with cholera toxin. Thus, lamina propria antitoxin-containing plasma cells appeared to be the source of protective antitoxin. However, after sequential s.c.-oral immunization with toxoid, protection against challenge with Vibrio cholerae far outlasted the major systemic and local antitoxin responses and was not obviously explained by either. These studies reveal methods for induction of a mucosal antitoxin response, but leave in question the mechanism of prolonged protection induced by s.c.-oral immunization of dogs.     

Food hypermotivation syndromes in rabbits with hyperactivity of lateral hypothalamic structures

A focus of hyperactivity in the initiating centers for food motivation was created in experiments on unrestrained rabbits by blocking inhibitory mechanisms in the lateral hypothalamus (LH) by injection of tetanus toxin (group 1) or by chronic electrical stimulation of the same region of the brain (group 2). The syndromes thus evoked differed in the intensity of food hypermotivation. The syndrome evoked by injection of tetanus toxin was characterized by very rapid development, clear symptoms of hyperphagia, and a severe course. Marked epileptiform activity was recorded in LH, evidence of the formation of a generator of pathologically enhanced excitation in the pacemaker of food motivation. Mechanisms of adequate correction of behavior depending on the result obtained were completely disturbed in the animals. The syndrome evoked by chronic electrical stimulation of LH was characterized by slower formation and a more favorable course. Relatively weak epileptiform activity was recorded in LH. Enhanced food-getting activity of the rabbits also was observed in the late periods after the beginning of electrical stimulation, but the animals' behavior did not lose its adaptive character in this case. The formation of the two syndromes of food hypermotivation described above was connected with the formation of two excitation generators, differing in the intensity of excitation produced by them.Laboratory of General Pathology of the Nervous System, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. Department of Pathological Physiology, Leningrad San.-Gig. Medical Institute. Group for Neurophysiology of Motivations, Institute of Normal Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 89, No. 3, pp. 281–284, March, 1980.     

Action of tetanus toxin on neuromuscular transmission

Summary Comparison of the effects produced by tetanus toxin and proserine (neostigmine) demonstrated in experiments on cats that tetanus toxin was not a cholinesterase inhibitor. There exists a functional antagonism between the action of the tetanus toxin and that of proserine. At the same time, functional synergism was found to exist between the action of the toxin and that of d-paracurarine as evidenced by the fact that curare block developed more rapidly in the toxin-poisoned muscle than in the normal one. With increase of intoxication periods there was a drop in the action potential amplitude in the muscle, and curare block developed more rapidly. A characteristic response of the intoxicated muscle to indirect rhythmic stimulation with a fairly high frequency (pessimal for the normal muscle) was an increase in the action potential during the initial stimuli, and a relative maintenance of its value during the later stimuli.The results of this work permit a conclusion that tetanus toxin disturbs the neuromuscular transmission. The possible mechanisms of this effect are discussed. It is very likely that tetanus toxin influences the excretion of the mediator by the presynaptic apparatus.Presented by V. V. Parin, Active Member, Academy of Medical Sciences, USSR Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 58, No. 10, pp. 65–70, October, 1964     

Pathogenetic factors of experimental photogenic epilepsy

The character of formation of an experimental syndrome of photogenic epilepsy, obtained by creating a generator of pathologically enhanced excitation in the lateral geniculate body by local injection of tetanus toxin, was studied in chronic experiments on rats. The initial manifestations appearing in the animals were shown to be due to pathological enhancement of specific sensory excitation in the lateral geniculate body and to be accompanied by a marked increase in amplitude of the evoked potential in the visual cortex. The subsequent development of a neuropathological syndrome was connected with diffuse disturbances of rhythmic electrical activity of the brain, characteristic of a state of increased epileptic predisposition in the experimental animals. The results of these experiments indicate an important role of specific and nonspecific factors of epileptogenesis in the formation of experimental photogenic epilepsy.Laboratory of General Pathology of the Nervous System, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 87, No. 1, pp. 8–12, January, 1979.