Utility of the Tétanos Quick Stick® in the vaccine catch-up of adult migrants without proof of prior vaccination

BackgroundUnknowing immunity status make migrants vaccine catch-up difficult. The interest of using a rapid tetanus immunotest as the Tétanos Quick Stick® (TQS®) to assess immunity status against tetanus has been evaluated in emergency rooms and it is now commonly used. The study aim was to evaluate TQS® as a tool for migrants’ vaccine catch-up.MethodsFrom December 2018 to February 2019, a prospective study was performed and included consecutively migrants who attented to the primary medicine outconsultation of a health care centre in Paris. Migrants above 18, without any records of tetanus immunization were included and a TQS® was performed during a medical consultation. Adapted vaccine catch-up was then proposed. Immunity against tetanus among migrants, factors associated with positive TQS® and costs savings were evaluated.ResultsTQS® test was positive for 32% of the 310 included patients. In the univariable analysis, factors associated to the presence of a positive TQS® test were a female gender (OR = 1.69 CI95% [1.02–2.80]) and an urban living in the country of origin (OR = 1.79 CI95% [1.07–3.02]). In the multivariable analysis, these factors were not significantly associated to a positive TQS®. Anamnesis was not correlated to the immunity status: only 26% of the migrants who reported vaccinations in childhood, adolescence and adulthood had a positive TQS® test. The use of TQS® test allowed savings of 6,522 US$ as compared to the immediate catch-up strategy for the 310 patients.ConclusionThe TQS® test is an acceptable, simple, rapid and cost saving test that could find a place in the migrants’ vaccine catch-up.     

Isolation and characterization of new human carrier peptides from two important vaccine immunogens

In the preparation of glycoconjugate vaccines in clinical practice, two highly immunogenic carrier proteins, CRM₁₉₇ and tetanus toxoid (TT), are predominantly conjugated with the capsular polysaccharides (CPSs) of bacterial pathogens. In addition, TT has long been used as an effective vaccine to prevent tetanus. While these carrier proteins play an important role in immunogenicity and vaccine design alike, their defined human major histocompatibility complex class II (MHCII) T cell epitopes are inadequately characterized. In this current work, we use mass spectrometry to identify the peptides from these carrier proteins that are naturally processed and presented by human B cells via MHCII pathway. The MHCII-presented peptides are screened for their T cell stimulation using primary CD4+ T cells from four healthy adult donors. These combined methods reveal a subset of eleven CD4+ T cell epitopes that proliferate and stimulate human T cells with diverse MHCII allelic repertoire. Six of these peptides stand out as potential immunodominant epitopes by responding in three or more donors. Additionally, we provide evidence of these natural epitopes eliciting more significant T cell responses in donors than previously published TT peptides selected from T cell epitope screening. This study serves toward understanding carrier protein immune responses and thus enables the use of these peptides in developing novel knowledge-based vaccines to combat persisting problems in glycoconjugate vaccine design.     

Tetanus incidence and immunity in Poland

The level of tetanus antibody was determined with the passive hemagglutination method in 503 sera of 10 to 90-year-old persons. Immunity to tetanus was age-dependent: the percentage of immune persons amounted to 90% –100% in persons below 40 years and then declined to 70% and 60% in persons in the 40 – 50 and above 50 year-old groups, respectively. Males above 30 years are better immunized than females. The comparison of the results of the present study with results of several periodic surveys performed in the last 27 years showed gradually increasing immunity level in all age groups. The findings are in agreement with age-dependent incidence of tetanus, which has become now, in Poland, a disease primarily of older people.     

Limited proteolysis of single-chain tetanus toxin by tissue enzymes,in cultured brain tissue and during retrograde axonal to the spinal cord

Summary Single-chain toxin was investigated in vitro and in vivo for limited proteolysis into the fully active two-chain toxin. Plasmin from serum, elastase and gelatinase from leucocytes, as well as clostripain from C. histolyticum cleaved single-chain toxin and increased by that way its ability to inhibit [³H]noradrenaline release in vitro. Cultured mouse brain generated fragments from ¹²⁵I-single-chain toxin which were cell-associated. Some of them comigrated in electrophoresis with light and heavy chain after mercaptolysis. When injected i. v. into rats, ¹²⁵I-single-chain-toxin disappeared from the blood with a half-life of about 11 h without signs of nicking. However, after its injection into the triceps surae muscle both single- and two-chain toxin were found in the ipsilateral ventral horn of the spinal cord. Thus single-chain toxin is subjected to limited proteolysis by enzymes involved in tissue damage, by cultured brain tissue, and during or after its retrograde axonal transport to the spinal cord. Limited proteolysis is necessary for the release of the light chain known to mediate the action of toxin on several systems.     

心肌营养素1/破伤风毒素重链C端片段融合蛋白的构建及其靶向大鼠嗜铬细胞瘤细胞的研究

目的：探讨心肌营养素1（cardiotrophin 1，CT1）/破伤风毒素重链C端片段（tentanus toxin C fragment,TTC）（CT1/TTC）融合蛋白的构建及其对大鼠嗜铬细胞瘤（pheochromocytoma,PC12）细胞的靶向性。方法：采用聚合酶链式反应（PCR）、T-A克隆等分子生物学方法构建CT1/TTC融合蛋白。体外培养PC12细胞,并将CT1/TTC融合蛋白与PC12细胞共培养,红色荧光免疫组化染色后在激光共聚焦显微镜下观察融合蛋白能否在TTC的靶向作用下进入PC12细胞。结果：成功构建了CT1/TTC融合蛋白,测序显示融合基因序列正确,免疫组化染色结果显示融合蛋白能够进入PC12细胞，并发出红色荧光。结论：采用PCR和T-A克隆等分子生物学方法能成功构建CT1/TTC融合蛋白．并且TTC能够将CT1靶向进入PC12细胞。     

Induction of antigen-specific isotype switching by in vitro immunization of human naive B lymphocytes

The use of in vitro immunization technology for the generation of human antigen-specific antibodies has essentially resulted in low affinity IgM antibodies, resembling an in vivo primary immune response. We now describe a detailed reproducible protocol for a two-step in vitro immunization, which yields isotype switched, antigen-specific human antibodies. The immunizing antigen was a 30aa synthetic peptide, containing both a B (15aa V3 peptide of the HIV-1) and a T helper cell epitope (15aa peptide from tetanus toxin). The immunization protocol includes: (i) a selection procedure of donors with a memory T cell response against tetatus toxoid; (ii) immunization of mature naive peripheral B lymphocytes in two distinct phases, involving a primary and a secondary step. None of the donors which were examined after primary 7immunization showed at any time an IgG anti-V3 specific antibody response, while all the donors showed an IgM response. After the secondary immunization step, anti-V3 antibodies of both IgM and IgG isotypes were detected. The switch frequency event was high among the tested donors (5/8).     

注射吸毒致破伤风的临床特点及分析

目的探讨注射吸毒致破伤风的诊断与治疗。方法回顾性分析了9例注射吸毒者破伤风的临床资料。结果9例注射吸毒致破伤风的病人中,5例治愈,4例死亡,死亡原因:3例因呼吸肌痉挛和分泌物过多阻塞呼吸道窒息死亡,1例因呼吸衰竭死亡。结论注射后皮肤和软组织感染可认为是引起破伤风外伤伤口。注射吸毒破伤风应按重症破伤风处理。及时气管切开或气管插管机械辅助呼吸可降低死亡率。     

Relations between the effect of tetanus toxin on the neuromuscular transmission and histological functional properties of various muscles of the rat

Summary Various doses of tetanus toxin were injected into three hind leg and two fore leg muscles of the rat. The neuromuscular transmission was tested by recording the mass action potential of the muscles elicited by a single electrical stimulus to the motor nerve after strong symptoms of local tetanus had developed. The muscle responses were depressed and blocked at lower toxin doses in the fast tibialis anterior than in the mixed gastrocnemius latemlis, while blocking of the slow soleus required the highest dose. The extensor carpi radialis and the flexor carpi ulnaris muscles showed medium sensitivity. In all five muscles the contraction time was measured and correlated with its individual minimal blocking dose. The more phasic (i.e., the faster) the muscle, the more sensitive its neuromuscular transmission was to tetanus toxin. The proportional distribution of red, white, and intermediate fibres, which are associated with specific end-plate types, was evaluated for the five muscles. The percentage of white fibres in the muscles displayed a very good negative correlation with the blocking dose. The relation between structures of end-plates and effects of tetanus toxin were analysed and it is suggested that the differences in sensitivity to tetanus toxin in the neuromuscular transmission in the five muscles is determined by a differential distribution of endplates with varying sensitivities to this toxin due to structural properties.This study is a part of a doctoral dissertation submitted by one of the authors (H.K.) to the Faculty of Medicine, University of Göttingen. Some of the results were presented at the 48th and 49th Congr. of German Physiol. Soc. (Kretzschmar et al., 1977, 1978) and at the 5th Internat. Conf. on Tetanus (Kretzschmar et al., 1979)     

Distinct sites of action of clostridial neurotoxins revealed by double-poisoning of mouse motor nerve terminals

(1) We investigated the effects of single- and double-poisoning with tetanus toxin (TeTx), botulinum neurotoxin type A (BoTx A) and botulinum neurotoxin type B (BoTx B) on spontaneous and nerve-evoked quantal transmitter release at motor endplates of the triangularis sterni preparation of the mouse. (2) Inhibitory effects of TeTx and BoTx B on spontaneous and nerve-evoked transmitter release were very similar, except that the action of BoTx B required 500-fold lower concentrations and was less dependent on temperature. BoTx A caused stronger inhibition of quantal release than TeTx or BoTx B, but was comparatively much easier counteracted by 4-aminopyridine (4-AP). (3) In contrast to BoTx A, with TeTx or BoTx B the increase of transmitter release following onset of 50 Hz nerve stimulation was delayed for a few seconds and synaptic latencies of quanta showed large variations. This release pattern was also evident in all double-poisoning experiments, regardless of intoxication sequence. (4) Inhibition of evoked release was found to be slightly stronger with TeTx than with BoTx B, so the amount of nerve-evoked quanta released after double-poisoning with any sequence of these toxins always approached that of TeTx. In no case supraadditive actions were observed. (5) A strong reduction of evoked quanta was observed when BoTx A was applied in addition to either of the two other toxins. With reversed poisoning sequences (BoTx A-TeTx or BoTx A-BoTx B) the resulting values remained at the extremely low level of BoTx A. (6) In the presence of 4-AP double-poisoning with any combination between BoTx A and TeTx or BoTx B (regardless of intoxication sequence) revealed supra-additive effects, since the number of quanta released was considerably lower than that obtained with any of the toxins alone (in the presence of 4-AP). (7) Our results indicate that tetanus toxin and botulinum toxin type B have a common site of action which is different and independent from that of botulinum toxin type A.This is part of the thesis of M. G. to be presented to the Fachbereich Humanmedizin, Justus-Liebig-Universität Gießen     

Analysis of tetanus toxin peptide/DR recognition by human T cell receptors reconstituted into a murine T cell hybridoma

We have previously reported that human T cell receptors (TcR) selected in the class II-restricted (HLA-DRB1*1302) response to a tetanus toxin peptide (tt830-843) frequently used the Vβ2 germ-line segment which paired with several Vα segments and that the putative CDR3 of both α and β chains showed remarkable heterogeneity. To analyze the structural basis for recognition of the tt830-843/DR complex, five of these TcR were reconstituted into a murine T cell hybridoma, 58 α^?β^?, by expressing the human α and β variable regions joined to the mouse α and β constant regions, respectively. The chimeric TcR, expressing the same Vβ germ-line segment (Vβ2), two expressing Vα21.1, twoVα17.1 and one Vα8.1 were shown to have the expected antigen specificity and DR restriction. Two lines of evidence suggested that the putative CDR3, although not conserved in these TcR, played a key role in recognition. First, two TcR with identical V germ-line segments but distinct CDR3 showed large differences in their capacity to react with the ligand. Second, interchanging the α and β chains from tt830-843/DR1302-specific TcR which differed in their CDR3 sequences invariably led to loss of recognition. We also asked whether germ-line Vα17.1 could functionally replace Vα21.1, as they appear to be related in their primary sequence. However, as in the case of CDR3 exchanges, Vα replacement abrogated TcR reactivity. Taken together, these data underline the fine interdependence of the structural components of the TcR binding site in defining a given specificity. Four of the TcR studied displaying promiscuous recognition were also tested against different DR alleles and site-directed mutants. The results of these experiments suggested that, in spite of their structural heterogeneity, anti-tt830-843 TcR may have a similar orientation with respect to the peptide/DR complex. The reconstitution system described herein should represent a valuable tool for detailed studies of human TcR specificity.