PURPOSE: Recombinant subunit vaccines provide safe and targeted protection against microbial infections. However, the protective efficacy of recombinant subunit vaccines tends to be less potent than the whole cell vaccines, especially when they are administered through mucosal routes. We have reported that a bacterial flagellin has strong mucosal adjuvant activity to induce protective immune responses. In this study, we tested whether FlaB could be used as a fusion partner of subunit vaccine for tetanus. MATERIALS AND METHODS: We constructed fusion proteins consisted with tetanus toxin fragment C (TTFC), the nontoxic C-terminal portion of tetanus toxin, and a Toll-like receptor 5 agonist from Vibrio vulnificus (FlaB). Mice were intranasally administered with fusion protein and protective immune responses of the vaccinated mice were analyzed. RESULTS: FlaB-TTFC recombinant protein induced strong tetanus-specific antibody responses in both systemic and mucosal compartments and prolonged the survival of mice after challenge with a supra-lethal dose of tetanus toxin. CONCLUSION: This study establishes FlaB as a successful fusion partner for recombinant subunit tetanus vaccine applicable through mucosal route, and it further endorses our previous observations that FlaB could be a stable adjuvant partner for mucosal vaccines.
Animals ; Antibody Formation ; Flagellin* ; Mice ; Tetanus ; Tetanus Toxin* ; Tetanus Toxoid ; Toll-Like Receptor 5 ; Vaccines ; Vaccines, Subunit ; Vibrio vulnificus

In the present study, we investigated the effect of Tetaus toxin (TeT) on cell proliferation and neuroblast differentiation using specific markers: 5-bromo-2-deoxyuridine (BrdU) as an exogenous marker for cell proliferation, Ki-67 as an endogenous marker for cell proliferation and doublecortin (DCX) as a marker for neuroblasts in the mouse hippocampal dentate gyrus (DG) after TeT treatment. Mice were intraperitoneally administered 2.5 and 10 ng/kg TeT and sacrificed 15 days after the treatment. In both the TeT-treated groups, no neuronal death occurred in any layers of the DG using neuronal nuclei (NeuN, a neuron nuclei maker) and Fluoro-Jade B (F-J B, a high-affinity fluorescent marker for the localization of neuronal degeneration). In addition, no significant change in glial activation in both the 2.5 and 10 ng/kg TeT-treated-groups was found by GFAP (a marker for astrocytes) and Iba-1 (a marker for microglia) immunohistochemistry. However, in the 2.5 ng/kg TeT-treated-group, the mean number of BrdU, Ki-67 and DCX immunoreactive cells, respectively, were apparently decreased compared to the control group, and the mean number of each in the 10 ng/kg TeT-treated-group was much more decreased. In addition, processes of DCX-immunoreactive cells, which projected into the molecular layer, were short compared to those in the control group. In brief, our present results show that low dosage (10 ng/kg) TeT treatment apparently decreased cell proliferation and neuroblast differentiation in the mouse hippocampal DG without distinct gliosis as well as any loss of adult neurons.
Adult ; Animals ; Bromodeoxyuridine ; Cell Proliferation ; Dentate Gyrus ; Exotoxins ; Fluoresceins ; Gliosis ; Humans ; Immunohistochemistry ; Mice ; Neurogenesis ; Neurons ; Tetanus ; Tetanus Toxin

Tetanus is a neurologic disorder caused by a tetanospasmin released from Clostridium tetani and usually occurs following a stab wound or dirty abrasion. Tetanus is uncommon in Korea due to the introduction of vaccination programs. Furthermore, tetanus associated with a gangrenous perforation of the small bowel is extremely rare. We report a case of tetanus developed in a patient who was diagnosed with a gangrenous perforation of the small bowel. This is the first reported case in Korea.
Clostridium tetani ; Digestive System Surgical Procedures ; Humans ; Intestinal Perforation ; Korea ; Metalloendopeptidases ; Nervous System Diseases ; Tetanus ; Tetanus Toxin ; Vaccination ; Wounds, Stab

Synthetic cannabinoids are one of most abused new psychoactive substances. The recreational use of abused drug has aroused serious concerns about the consequences of these drugs on infection. However, the effects of synthetic cannabinoid on resistance to tetanus toxin are not fully understood yet. In the present study, we aimed to determine if the administration of synthetic cannabinoids increase the susceptibility to tetanus toxin-induced motor behavioral deficit and functional changes in cerebellar neurons in mice. Furthermore, we measured T lymphocytes marker levels, such as CD8 and CD4 which against tetanus toxin. JWH-210 administration decreased expression levels of T cell activators including cluster of differentiation (CD) 3ε, CD3γ, CD74p31, and CD74p41. In addition, we demonstrated that JWH-210 induced motor impairment and decrement of vesicle-associated membrane proteins 2 levels in the cerebellum of mice treated with tetanus toxin. Furthermore, cerebellar glutamatergic neuronal homeostasis was hampered by JWH-210 administration, as evidenced by increased glutamate concentration levels in the cerebellum. These results suggest that JWH-210 may increase the vulnerability to tetanus toxin via the regulation of immune function.
Animals ; Cannabinoids ; Cerebellar Diseases* ; Cerebellum ; Glutamic Acid ; Homeostasis ; Immunosuppression* ; Mice* ; Neurons ; R-SNARE Proteins ; T-Lymphocytes ; Tetanus ; Tetanus Toxin

PURPOSE: We conducted the study to evaluate the immunogenicity and safety of three component DTaP vaccine (Infanrix(R)) in a group of Korean healthy infants on a three-dose primary vaccination. And we compared the immunogenicity of this DTaP vaccine with two component DTaP vaccine which has been widely used in Korea. METHODS: We enrolled one hundred fifty one healthy infants aged 8-9 weeks. These infants were vaccinated at age 2, 4 and 6 months of age with three component DTaP vaccine. Solicited adverse events were actively monitored for 72 hours following each vaccination, and all adverse events after each vaccination were observed for three weeks. Anti-diphtheria toxoid Ab., anti-tetanus toxoid Ab., anti-pertussis toxin Ab., anti-filamentous hemagglutinin Ab., and anti-pertactin Ab. were measured using ELISA for assessing immunogenicity of study vaccine in 60 infants. Immunogenicity analysis of two component DTaP vaccine was performed with same methods in 14 infants as control. RESULTS: The seroconversion rates of anti-diphtheria toxoid Ab, anti-tetanus toxoid Ab. anti- filamentous hemagglutinin Ab. were 100% in both group. Seroconversion rate of anti-pertactin Ab in study group was 100%, but the rate in control group was 50%. However, geometric mean concentration of anti-pertussis toxin Ab. was higher in control group. Mild local and systemic reactions were observed within three days after vaccination, and no serious adverse events related study vaccine were happened during study period. CONCLUSIONS: Our study results suggest that three component DTaP vaccine (Infanrix(R)) is a well- tolerable and high immunogenic vaccine, especially anti-Pertactin Ab. of the study vaccine is very immunogenic. It can be available as routine DTaP vaccination in our infants.
Diphtheria-Tetanus-acellular Pertussis Vaccines* ; Enzyme-Linked Immunosorbent Assay ; Hemagglutinins ; Humans ; Infant* ; Korea ; Pertussis Toxin ; Vaccination

Tetanus is a neurologic disease which features the muscle spasm as the hallmark. It is an infectious disease with high mortality rate triggered by tetanospasmin produced by Clostridium tetani. This report concerns incidence of tetanus consequent to oriental medical care such as acupuncture and moxibustion. Although the tetanus occurrence has shown a remarkable decline since nationwide vaccinations in some of the developed countries, including Korea, it still remains to be an important issue, to be dealt within Korea, as the majority of the patients are old aged and Korean population is rapidly becoming an aging society. Furthermore, since more elders are coming to rely on Oriental medicine in Korea, the Korean elders are at a higher risk than elsewhere. The lack of medical experiences, including those in oriental medical field, has been hindering early diagnosis of Tetanus. This study aims to encourage rapid and accurate decisions in diagnosis and treatment through reviewing symptoms particularly specific to tetanus, and also to arouse attention to the riskiness of invasive procedures involving skin puncture.
Acupuncture ; Aged ; Aging ; Clostridium tetani ; Communicable Diseases ; Developed Countries ; Early Diagnosis ; Humans ; Incidence ; Korea ; Medicine, East Asian Traditional ; Metalloendopeptidases ; Moxibustion ; Punctures ; Skin ; Spasm ; Tetanus ; Tetanus Toxin ; Vaccination

BACKGROUND: Tetanus toxin selectively blocks inhibitory synapses in the brainstem as well as the spinal cord. Therefore, in contradiction to Stiff Person syndrome, patients with generalized tetanus usually show abnormal masseter silent periods as well as abnormal F/M amplitude or H/M amplitude ratios. This study aimed to verify the characteristics of electrophysiological findings of generalized tetanus. METHODS: The authors retrospectively studied clinical and electrophysiological characteristics of 7 patients with generalized tetanus, who were admitted to the neurology department of Hallym Medical Center from 1995 to 2005. RESULTS: All the seven patients showed abnormal masseter silent periods. Three of them showed somewhat improvement in the silent period at follow-up study as trismus was improving. Full NCSs done in two patients did not show any abnormalities except an increased F/M amplitude ratio. One patient with a wound site in his left finger showed an abnormal F/M amplitude ratio only in the right upper extremity without involvement of other extremities. Another patient showed an increased H/M amplitude ratio without an increased F/M amplitude ratio. (In this patient we did not conduct full NCS tests.) CONCLUSIONS: The Masseter silent period could be used as a diagnostic tool and parameter of clinical improvement in patients with generalized tetanus.
Brain Stem ; Extremities ; Fingers ; Follow-Up Studies ; H-Reflex ; Humans ; Neurology ; Retrospective Studies ; Spinal Cord ; Stiff-Person Syndrome ; Synapses ; Tetanus Toxin ; Tetanus* ; Trismus ; Upper Extremity ; Wounds and Injuries

Recently we have shown that a bacterial flagellin, Vibrio vulnifiucs FlaB (Vv-FlaB), has a strong adjuvant activity to induce protective immune response. In order to investigate the adjuvanticity of Vv-FlaB, we prepared highly purified recombinant protein by using an intein fusion protein purification system. However, in the process of the purification, we unexpectedly encountered a contamination with a 70 kDa protein. We proved the 70 kDa protein as the heat shock protein 70 (HSP70) by Western blotting. Unfortunately, it was reported that the HSP70 has a strong adjuvanticity. In this study we investigated the role of contaminating HSP70 on the Vv-FlaB-mediated adjuvanticity. We separated Vv-FlaB and HSP70 by using a high performance protein purification chromatography and compared adjuvant activities of Vv-FlaB, HSP70 and Vv-FlaB/HSP70 mixture. Using an intranasal immunization mouse model, we observed that co-administration of the flagellin with tetanus toxoid (TT) induced significantly enhanced TT-specific antibody (Ig) responses. However contaminating doses of HSP70 did not affect the adjuvanticity of Vv-FlaB and furthermore HSP70 alone did not enhance TT-specific Ig response and protective immunity against lethal challenge with tetanus toxin. These results show that the HSP70 contaminating Vv-FlaB preparations did not affect the adjuvanticity of Vv-FlaB.
Animals ; Blotting, Western ; Chromatography ; Flagellin* ; Heat-Shock Proteins* ; Hot Temperature* ; HSP70 Heat-Shock Proteins* ; Immunization ; Inteins ; Mice ; Staphylococcal Protein A ; Tetanus Toxin ; Tetanus Toxoid ; Vibrio vulnificus ; Vibrio*

BACKGROUND: The incidence of tetanus in adults and neonatal tetanus have been markedly reduced by world-wide use of DTP vaccines. But, tetanus is still one kind of major health problems in many developing countries, and several serosurvey stduies in developed countries revealed that substantial proportions of adult population may lack immunity against tetanus and immunity level against tetanus is continuously decreasing by age. In Korea, tetanus outbreaks have been disappeared since the 1980s by high acceptant DTaP vaccination rates. Annually, few tetanus patient has been reported since 1990s. But, there have been no seroepidemiological studies to tetanus, no trials to assess tetanus immunizations. And we do not use Td vaccine in adults for maintaning tetanus immunity. In this aspect, we conducted age related survey of immunity to tetanus and indirectly assessed the immunogenecity of tetanus vaccines, used in Korea. METHODS: For the evaluation of age related serosurvey of tetanus immunity in Korean population, study subjects were classified into 16 groups (A~J group; below 10 years with one year interval, K~O group; 11~60 yrs with 10 years interval, P group; over 60 yrs). The numbers of each group were 100, and sex distributions of each group were almostly equal. And for the indirect assessment of tetanus immunization in Korean children, children under 15 years old age were classified into 6 groups (I~VI) according to the status of DTaP vaccination. The numbers of this each group were 50, and sex ratio was almostly equal. Specific IgG antibody to tetanus toxin were detected by ELISA. And the ANOVA repeated t-test was used to compare antibody levels in study groups. RESULTS: In age related groups, the antibody levels to tetanus toxin were well maintained until 20 years old age group (L group), but thereafter the titers abruptly decreased below 0.1 IU/mL and over 75% populations among the groups over 30 years old age needed maintenance of protective immunity to tetanus. The antibody level of male was statistically higher than that of female in P group. In the groups related DTaP vaccination status, the antibody titer was very low in prevaccination group (I), but the titers after primary vaccinations were sharply increased and highly maintained until 15 years. CONCLUSOIN: The results of our study revealed that the immunity to tetanus was dramatically decreased in age groups over 30 years old. This result indicates that Td vaccination program in adults should be considered for maintenance of immunity to tetanus. And our study indicate that DTaP vaccination programs and vaccines, used in Korea, are effective for acquisition and maintenance of tetanus immunity in Korean children.
Adolescent ; Adult ; Child ; Developed Countries ; Developing Countries ; Disease Outbreaks ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunization ; Immunoglobulin G ; Incidence ; Korea ; Male ; Seroepidemiologic Studies ; Sex Distribution ; Sex Ratio ; Tetanus Toxin ; Tetanus* ; Vaccination ; Vaccines ; Young Adult

OBJECTIVETo obtain highly purified tetanus toxin fragment C (TTC) with good immunogenicity.

METHODSThe gene fragment encoding TTC was amplified from Clostridium tetani plasmid DNA by PCR, inserted into the vector pET43.1a (+) and expressed in E. coli BL21(DE3)plysS. After purification using Ni2+-chelate affinity chromatography, the expressed fusion protein was digested by thrombin and the resultant TTC protein was purified with Ni2+-chelate affinity chromatography followed by identification with SDS-PAGE and Western blotting. The purifed TTC protein was then used to immunize mice to test its immunogenecity.

RESULTSThe 1373-bp gene fragment encoding TTC was obtained, and the constructed recombinant expression vector pET43.1a (+)-TTC was successfully expressed in E. coli BL21(DE3)plysS. SDS-PAGE identified a recombinant fusion protein with relative molecular mass (Mr) of 117 000, which accounted for 22% of the total bacterial protein. The TTC protein with Mr of 50 000 was obtained after purification of the thrombin digestion products of the fusion protein, with a purity reaching 95.5%. Both the fusion protein and TTC protein could be recognized by anti-tetanus toxin antibody as shown by Western blotting. The titer of the anti-serum from mice immunized with the TTC protein was 1:25 600, and the anti-serum could specifically bind to tetanus toxin.

CONCLUSIONHighly purified and immunogenetic TTC protein has been successfully obtained, which provides a good model antigen for studying antigen presentation and immune responses in vivo.

Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Peptide Fragments ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Tetanus Toxin ; biosynthesis ; genetics ; immunology ; Tetanus Toxoid ; immunology