Tokyo Strain Research Articles

Comparative transcriptomics reveals common and strain-specific responses of human macrophages to infection with Mycobacterium tuberculosis and Mycobacterium bovis BCG

Mycobacterium tuberculosis (MTB) and Mycobacterium bovis (M. bovis) are closely related pathogenic mycobacteria known to cause chronic pulmonary infections in both humans and animals. Despite sharing nearly identical genomes and virulence factors, these two bacteria display variations in host tropism, epidemiology, and clinical presentations. M. bovis Bacillus Calmette–Guérin (BCG) is an attenuated strain of M. bovis commonly utilized as a vaccine for tuberculosis (TB). Nevertheless, the molecular underpinnings of these distinctions and the intricacies of host-pathogen interactions remain areas of ongoing research. In this study, a comparative transcriptomic analysis was conducted on human leukemia macrophages (THP-1) infected with either MTB H37Rv or M. bovis BCG (Tokyo strain) to elucidate common and strain-specific responses at the transcriptional level. RNA sequencing was utilized to characterize the transcriptomes of human primary macrophages infected with MTB or BCG at 6 and 24 h post-infection. The findings indicate that both MTB and BCG induce substantial and dynamic alterations in the transcriptomes of THP-1, with a notable overlap in the quantity and extent of differentially expressed genes (DEGs). Moreover, gene ontology (GO) enrichment analysis unveiled shared pathways related to immune response, cytokine signaling, and apoptosis. The immune response of macrophages to bacterial infections at 6 h exhibited significantly greater intensity compared to that at 24 h. Furthermore, distinct gene sets displaying notable variances between MTB and BCG infections were identified. The profound impact of MTB infection on macrophage gene expression, particularly within the initial 6 h, was evident. Additionally, downregulation of pathways such as Focal adhesion, Rap1 signaling pathway, and Regulation of actin cytoskeleton was observed. The pathways associated with inflammation reactions and cell apoptosis exhibited significant differences, with BCG triggering macrophage apoptosis and MTB enhancing the survival of intracellular bacteria. Our findings reveal that MTB and BCG provoke similar yet distinct transcriptional responses in human macrophages, indicating variations in their pathogenesis and ability to adapt to host environments. These results offer novel insights into the molecular mechanisms governing host-pathogen interactions and may contribute to a deeper understanding of TB pathogenesis.

Genetic engineering employing MPB70 and its promoter enables efficient secretion and expression of foreign antigen in bacillus Calmette Guérin (BCG) Tokyo.

Vaccination is an important factor in public health. The recombinant bacillus Calmette Guérin (rBCG) vaccine, which expresses foreign antigens, is expected to be a superior vaccine against infectious diseases. Here, we report a new recombination platform in which the BCG Tokyo strain is transformed with nucleotide sequences encoding foreign protein fused with the MPB70 immunogenic protein precursor. By RNA-sequencing, mpb70 was found to be the most transcribed among all known genes of BCG Tokyo. Small oligopeptide, namely, polyhistidine tag, was able to be expressed in and secreted from rBCG through a process in which polyhistidine tag fused with intact MPB70 were transcribed by an mpb70 promoter. This methodology was applied to develop an rBCG expressing the receptor binding domain (RBD) of severe acute respiratory syndrome coronavirus 2. Immunoblotting images and mass spectrometry data showed that RBD was also secreted from rBCG. Sera from mice vaccinated with the rBCG showed a tendency of weak neutralizing capacity. The secretion was retained even after a freeze-drying process. The freeze-dried rBCG was administered to and recovered from mice. Recovered rBCG kept secreting RBD. Collectively, our recombination platform offers stable secretion of foreign antigens and can be applied to the development of practical rBCGs.

Peripheral and Central Nervous System Immune Effects of Bacillus Calmette‐Guérin (BCG) Vaccination in older adults with and without cognitive impairment

AbstractBackgroundThe live‐attenuated vaccine Bacillus Calmette‐Guérin (BCG), is widely used for the prevention of tuberculosis, and has myriad non‐specific beneficial effects in T1DM, multiple sclerosis, decreasing all‐cause childhood mortality, etc. Alzheimer’s Disease (AD) is a complex neurological disease. The innate immune system, intimately involved in the early and late phases of AD, may prove an ideal target for mono‐ or multi‐pronged therapy against AD. Data from multiple population health studies find that BCG treatment for bladder cancer is linked with decreased incidence of AD. Preclinical data also supports an AD‐beneficial effect of the vaccine. While mechanisms of some of these non‐specific vaccine effects have been elucidated, the mechanism behind BCG’s putative central nervous system (CNS) benefits is unclear. Here, we conducted a pilot open‐label study of the BCG (Tokyo strain) in the older adults to study CNS beneficial effects, particularly as they pertain to AD.MethodsBaseline measures of participants (N = 16) with or without mild cognitive impairment (MCI) included cognitive and neuropsychological testing, phlebotomy, and lumbar puncture (LP), followed by an initial intradermal BCG, with a booster given 1mo later. Neuropsychological testing, phlebotomy and LP were repeated at 3mo, 6mo (no LP), and 12mo. The primary endpoints include 1) the safety of the vaccine in older adults and 2) target engagement. Secondary and exploratory endpoints include 1) functional, immunological, and AD‐related biomarker measures and 2) cognitive and neuropsychological effects.ResultsOur interim analysis demonstrated an overall improvement in repeatable battery for neuropsychological status (RBANS) total score index (TSI), notably in subsections of delayed recall, and visuospatial and constructional indices. Our results also show robust peripheral target engagement and the induction of trained immunity in the peripheral blood cells. We will further show the effects on various AD‐related biomarkers in plasma and cerebrospinal fluid.ConclusionThese findings support that the BCG vaccine is safe in older adults and may improve cognition (enrollment of a placebo control group is underway). A further study including subjects with MCI and AD is ongoing. Our interventional studies with BCG begin to explore whether this strategy of retraining the neuroimmune axis can prevent and/or treat AD.

A Territory-wide Study Investigating the Dose and Efficacy of Different Bacillus Calmette-Guérin Strains in Patients with Intermediate- and High-risk Non–muscle-invasive Bladder Cancer

BackgroundCurrent European Association of Urology (EAU) guidelines support adjuvant intravesical Bacillus Calmette-Guérin (BCG) treatment after Transurethral Resection of Bladder Tumor (TURB) for intermediate- or high-risk Non–Muscle-Invasive Bladder Cancer (NMIBC) patients, aiming to reduce the risk of tumor recurrence. The quality of data, however, does not allow definitive conclusions on whether different strains and dosages of BCG have different efficacies on long-term survival outcomes. ObjectiveTo evaluate the long-term survival outcomes of different strains and dosages of BCG in patients with NMIBC. Design, setting, and participantsAll NMIBC patients treated with intravesical BCG therapy from 2001 to 2020 were identified using a territory-wide database in Hong Kong. InterventionBCG strains and dosages (Connaught strain 81 mg, Connaught strain 27 mg, Tokyo strain 80 mg, and Danish strain 30 mg) were retrieved from medical records. Outcome measurements and statistical analysisOverall Survival (OS), Cancer-Specific Survival (CSS), Recurrence-Free Survival (RFS), and Progression-Free Survival (PFS) were analyzed using the Kaplan-Meier method. A multivariable Cox regression analysis was used to adjust potential confounding factors, and to estimate Hazard Ratio (HR) and 95% confidence interval (CI) of different BCG strains. A further subgroup analysis on adequate versus inadequate BCG treatment was performed. Results and limitationsA total of 2602 NMIBC patients treated with intravesical BCG were identified. Among them, 1291 (49.6%) received Connaught strain 81 mg, 199 (7.6%) received Connaught strain 27 mg, 1014 (39.0%) received Tokyo strain, and 98 (3.8%) received Danish strain. The median follow-up was 11.0 years. No statistically significant differences in OS, CSS, RFS, and PFS were detected among the different groups. At the multivariable analysis, the Connaught strain 27 mg group was inferior to the Connaught strain 81 mg group in terms of OS (HR: 1.26, 95% CI: 1.05–1.51), CSS (HR: 1.69, 95% CI: 1.08–2.66), and PFS (HR: 1.86, 95% CI: 1.20–2.88). Adequate BCG treatment was associated with improved OS (HR: 0.82, 95% CI: 0.73–0.92), CSS (HR: 0.64, 95% CI: 0.47–0.86), RFS (HR: 0.80, 95% CI: 0.70–0.92), and PFS (HR: 0.52, 95% CI: 0.39–0.68). Among patients treated with adequate BCG, at the multivariable analysis the Connaught strain 27 mg group showed worse results than the Connaught strain 81 mg group in terms of CSS (HR: 1.93, 95% CI: 1.07–3.51). Compared with the Connaught strain 81 mg group, both Tokyo and Danish strains had similar survival outcomes in the whole cohort and the adequate BCG treatment subgroup. ConclusionsOur findings suggest that adequate BCG remains the most important factor in optimizing survival outcomes in patients with intermediate- and high-risk NMIBC. No significant differences in survival outcomes were observed between full-dose Connaught, Tokyo, and Danish strains. Reduced-dose Connaught strain was associated with the worst survival outcomes. Patient summaryWe evaluated the efficacy of different strains and dosages of bacillus Calmette-Guérin (BCG) in patients with intermediate- or high-risk non–muscle-invasive bladder cancer in the past two decades in Hong Kong. We conclude no significant differences in long-term survival outcomes in terms of full-dose Connaught, Tokyo, and Danish strains, while reduced-dose Connaught strain was inferior to the full-dose group. Adequate BCG treatment benefits long-term survival.

MP08-17 BACILLUS CALMETTE–GUÉRIN STRAINS DON’T MATTER: RESULTS FROM A TERRITORY-WIDE RETROSPECTIVE COHORT STUDY

You have accessJournal of UrologyCME1 Apr 2023MP08-17 BACILLUS CALMETTE–GUÉRIN STRAINS DON’T MATTER: RESULTS FROM A TERRITORY-WIDE RETROSPECTIVE COHORT STUDY Kang Liu, Hongda Zhao, Xuan Chen, Peter Ka-Fung Chiu, Chi-Fai Ng, and Jeremy Yuen-Chun Teoh Kang LiuKang Liu More articles by this author , Hongda ZhaoHongda Zhao More articles by this author , Xuan ChenXuan Chen More articles by this author , Peter Ka-Fung ChiuPeter Ka-Fung Chiu More articles by this author , Chi-Fai NgChi-Fai Ng More articles by this author , and Jeremy Yuen-Chun TeohJeremy Yuen-Chun Teoh More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003223.17AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Adjuvant intravesical Bacillus Calmette–Guérin (BCG) immunotherapy is recommended for non-muscle-invasive bladder cancer (NMIBC), except those low-risk patients. However, whether different strains of BCG have different efficacy on survival outcomes in the long term remains controversial. The current study tries to provide further evidence with a territory-wide retrospective cohort. METHODS: We retrospectively included 2,602 NMIBC patients with adjuvant intravesical BCG immunotherapy around Hong Kong from 2001 to 2020. Kaplan–Meier plotter with the log-rank test was used to explore overall survival (OS), cancer-specific survival (CSS), recurrence-free survival (RFS), and progression-free survival (PFS). Multivariable Cox regression analysis was used to balance clinical characteristics and to estimate hazard ratios (HRs) with a 95% confidence interval (CI). Adequate BCG treatment subgroup analysis was performed. RESULTS: Of 2,602 patients, 1291 (49.6%) received Connaught strain 81mg, 199 (7.6%) received Connaught strain 27mg, 1014 (49.2%) received Tokyo strain, and 98 (3.8%) received Danish strain. Although the Connaught strain 27mg group were inferior to the Connaught strain 81mg group in OS (HRs: 1.256, 95% CI: 1.047-1.507), CSS (HRs: 1.693, 95% CI: 1.079-2.657) and PFS (HRs: 1.857, 95% CI: 1.198-2.876) in the whole cohort, the differences were eliminated upon subgroup analysis focusing on patients who received adequate BCG treatment (Table 1, Figure 1). The other two strains, i.e. Tokyo and Danish strains, had similar survival outcomes with the Connaught strain 81mg group in both the whole cohort and the adequate BCG treatment subgroup. CONCLUSIONS: In this large BCG cohort, no significant differences in survival outcomes were observed between Connaught, Tokyo, and Danish strains as soon as adequate BCG treatment was given. Source of Funding: UGC Research Matching Fund 8601521 © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e101 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kang Liu More articles by this author Hongda Zhao More articles by this author Xuan Chen More articles by this author Peter Ka-Fung Chiu More articles by this author Chi-Fai Ng More articles by this author Jeremy Yuen-Chun Teoh More articles by this author Expand All Advertisement PDF downloadLoading ...

BCG Vaccine-associated Complications in a Large Cohort of Children With Combined Immunodeficiencies Affecting Cellular and Humoral Immunity.

To present the details of Bacillus Calmette-Guérin (BCG)-vaccine associated complications (VACs) in combined immunodeficiencies (CID) patients. Five centers participated in this retrospective study and completed a data form, which included general patients' information, clinical and laboratory data. Among 236 CID patients, 127 were BCG vaccinated. 41.9% of patients with family history of CID and 17.1% who were diagnosed by screening were BCG vaccinated. Twenty-three patients (18.1%) developed BCG-VACs. The median age of VACs was 6 months and the median time from vaccination to complications was 6 months. The highest rate of BCG-VACs was recorded in patients receiving the Russian BCG strain compared to the Tokyo and Danish strains. Univariate analysis of T-lymphocyte subsets showed increased odds of BCG complications in patients with CD3+, CD4+, and CD8+ counts of ≤250 cells/µL. Only CD8 + count ≤250 cells/µL had increased such odds on multivariate analysis. VACs were disseminated in 13 and localized in 10 patients. Localized complication occurred earlier after vaccination (median: 4 months) compared with disseminated ones (median: 7 months). There were no significant associations between sex, administered vaccine strain, serum immunoglobulins levels, lymphocyte subsets counts, and the chance of having either localized or disseminated BCG-related complications. Although contraindicated, many patients with CID continue to be vaccinated with BCG. Low CD8 + count is a risk factor for BCG-related complications and localized complications occurred earlier than disseminated ones. Considerations should be undertaken by health care authorities especially in countries with high incidence of CID to implement newborn screening, delay the time of BCG vaccine administration beyond 6 months of age and to use the relatively safer strains like the Danish and Tokyo ones.

Selective delipidation of Mycobacterium bovis BCG retains antitumor efficacy against non-muscle invasive bladder cancer.

Repeated instillations of bacillus Calmette et Guérin (BCG) are the gold standard immunotherapeutic treatment for reducing recurrence for patients with high-grade papillary non-muscle invasive bladder cancer (NMIBC) and for eradicating bladder carcinoma-in situ. Unfortunately, some patients are unable to tolerate BCG due to treatment-associated toxicity and bladder removal is sometimes performed for BCG-intolerance. Prior studies suggest that selectively delipidated BCG (dBCG) improves tolerability of intrapulmonary delivery reducing tissue damage and increasing efficacy in preventing Mycobacterium tuberculosis infection in mice. To address the lack of treatment options for NMIBC with BCG-intolerance, we examined if selective delipidation would compromise BCG's antitumor efficacy and at the same time increase tolerability to the treatment. Murine syngeneic MB49 bladder cancer models and in vitro human innate effector cell cytotoxicity assays were used to evaluate efficacy and immune impact of selective delipidation in Tokyo and TICE BCG strains. Both dBCG-Tokyo and dBCG-TICE effectively treated subcutaneous MB49 tumors in mice and enhanced tumor-infiltrating CD8+ T and natural killer cells, similar to conventional BCG. However, when compared to conventional BCG, only dBCG-Tokyo retained a significant effect on intratumoral tumor-specific CD8+ and γδ T cells by increasing their frequencies in tumor tissue and their production of antitumoral function-related cytokines, i.e., IFN-γ and granzyme B. Further, dBCG-Tokyo but not dBCG-TICE enhanced the function and cytotoxicity of innate effector cells against human bladder cancer T24 in vitro. These data support clinical investigation of dBCG-Tokyo as a treatment for patients with BCG-intolerant NMIBC.

Impact of maintenance therapy using a half dose of the bacillus Calmette\u2013Gu\xe9rin Tokyo strain on recurrence of intermediate and high-risk nonmuscle invasive bladder cancer: a retrospective single-center study

BackgroundData are scarce regarding intravesical maintenance therapy (MT) with the low-dose bacillus Calmette–Guérin (BCG) Tokyo strain. We investigated the efficacy and safety of MT with a half dose of the Tokyo strain for patients following transurethral resection of nonmuscle invasive bladder cancer (NMIBC).MethodsThis study retrospectively reviewed clinical data on 78 patients diagnosed with intermediate or high-risk NMIBC followed by either MT (n = 38) or IT alone (n = 40) between January 2012 and March 2018. Statistical analysis was performed to compare recurrence-free survival (RFS) and adverse effects between the two groups. BCG was instilled once weekly for 6 weeks as IT, then once weekly in 2-week for a total of 20 instillations over 3 years.ResultsKaplan–Meier analyses showed that patients undergoing MT had significantly better RFS than did those undergoing IT alone (hazard ratio (HR):0.32, 95% confidence interval (CI):0.12–0.89, P = 0.02). The 3-year RFS was 65.0% in the IT group and 89.5% in the MT group. Multivariate analysis showed that MT was associated with a reduced risk of recurrence (HR: 0.32, 95% CI:0.11–0.93, P = 0.03). One MT patient (2.6%) exhibited progression.ConclusionsThe BCG Tokyo strain showed acceptable efficacy and safety in patients undergoing MT; thus, it is a potential treatment for preventing bladder cancer recurrence.

Vaccination with Intradermal Bacillus Calmette-Guérin Provides Robust Protection against Extrapulmonary Tuberculosis but Not Pulmonary Infection in Cynomolgus Macaques.

Recently, the efficacy of Mycobacterium bovis bacillus Calmette-Guérin (BCG) vaccination is being reassessed in accordance with the achievements of clinical tuberculosis (TB) vaccine research. However, the mechanisms ultimately determining the success or failure of BCG vaccination to prevent pulmonary TB remain poorly understood. In this study, we analyzed the protective effects of intradermal BCG vaccination by using specific pathogen-free cynomolgus macaques of Asian origin that were intradermally vaccinated with BCG (Tokyo strain) followed by Mycobacterium tuberculosis (Erdman strain) infection. Intradermal BCG administration generated TB Ag-specific multifunctional CD4 T cell responses in peripheral blood and bronchoalveolar lavage and almost completely protected against the development of TB pathogenesis with aggravation of clinical parameters and high levels of bacterial burdens in extrapulmonary organs. However, interestingly, there were no differences in bacterial quantitation and pathology of extensive granulomas in the lungs between BCG-vaccinated monkeys and control animals. These results indicated that the changes in clinical parameters, immunological responses, and quantitative gross pathology that are used routinely to determine the efficacy of TB vaccines in nonhuman primate models might not correlate with the bacterial burden and histopathological score in the lung as measured in this study.

Investigation of adverse events following bacille Calmette-Guérin immunization using immunization safety surveillance system in Korea Centers for Disease Control and Prevention.

PurposeThe report of adverse events following immunization (AEFI) in Korea has continued since 1994, and the most frequently reported cases of AEFI of Korea Centers for Disease Control and Prevention (KCDC) is bacille Calmette-Guérin (BCG). Meanwhile, various inoculation methods and strains have been used in the past 6 years in Korea. Therefore, we investigated AEFI of BCG by strain types and inoculation methods using immunization safety surveillance of KCDC.Materials and MethodsWe reviewed BCG AEFIs registered in the KCDC from January 2013 to June 2018.ResultsThere were 336 AEFI cases during the period, and average time interval from vaccination to symptom onset was within 2 months. AEFI proportion was 6.4 cases per 100,000 doses for BCG percutaneous Tokyo strain, 41.6 cases per 100,000 doses of BCG intradermal Danish strain, and 25.9 cases per 100,000 doses of BCG intradermal Tokyo strain. Intradermal type was more reported AEFI than percutaneous type in the same strain. The most common adverse events were local reaction like BCG lymphadenitis and severe adverse reactions such as osteomyelitis or disseminated BCG disease were 0.1 to 0.2 cases per 100,000 doses which are correlated with the range of World Health Organization published AEFI rates.ConclusionThe AEFI reporting rate does not equal the actual proportion of AEFI occurrence. Because AEFI monitoring is a passive surveillance system, various factors might influence the number of events reported. Nevertheless, it is important to analyze BCG AEFI by vaccine strains and inoculation method using surveillance data of KCDC.

Inhibition of sodium-glucose cotransporter 2 ameliorates renal injury in a novel medaka model of nonalcoholic steatohepatitis-related kidney disease.

The effect of sodium‐glucose cotransporter 2 inhibitor (SGLT2I) on nonalcoholic steatohepatitis (NASH) has been reported, but there are few studies on its effect on NASH‐related renal injury. In this study, we examined the effect of SGLT2I using a novel medaka fish model of NASH‐related kidney disease, which was developed by feeding the d‐rR/Tokyo strain a high‐fat diet. SGLT2I was administered by dissolving it in water of the feeding tank. SGLT2I ameliorates macrophage accumulation and oxidative stress and maintained mitochondrial function in the kidney. The results demonstrate the effect of SGLT2I on NASH‐related renal injury and the usefulness of this novel animal model for research into NASH‐related complications.

Comparison of pellicle and shake flask-grown BCG strains by quality control assays and protection studies

A global BCG vaccine shortage began in 2013 which impacted availability for infant vaccinations, as well as preclinical studies and clinical trials of new TB vaccines. Stakeholders met in 2015 at McGill University in Montreal to discuss the shortage and potential mitigation strategies. Manufacturing BCG through a more tractable liquid fermentation process instead of the traditional pellicle growth method was considered a potentially viable strategy. This pilot program compared pellicle-grown and shake flask-grown BCG strains (as a first step towards modeling fermenter-produced BCG vaccine) in selected quality control assays, as well as mouse and guinea pig protection studies. Conventional pellicle-grown, lyophilized BCG WHO Reference Reagents (Danish, Moreau, Russian, Tokyo strains) were obtained from the National Institute for Biological Standards and Control (NIBSC), UK. Strains were grown in shake flasks and glycerol stocks prepared. Shake flask-grown BCG culture preparations generally met the requirements of quality control testing at NIBSC. In mouse and guinea pig protection studies there were no significant differences in lung colony forming units (CFUs) between shake flask-grown and pellicle-grown preparations, with the exception of BCG Russian, where the shake flask-grown preparation protected better in mice (P = 0.0042), but the pellicle-grown preparation protected better in guinea pigs (P = 0.0015). Producing BCG vaccines by a more tractable liquid growth process could be a viable solution to the global BCG shortage.

Post-Operative Immediate Single Instillation of Chemotherapy as Prevention of Recurrence after Transurethral Resection of Intermediate-High Risk Non-Muscle-Invasive Bladder Cancer

We retrospectively evaluated the efficacy of immediate single instillation (SI) of pirarubicine hydrochloride (THP) in the chemoprevention of intermediate and high risk patients with non-muscle-invasive bladder cancer (NMIBC). The study population consisted of 256 intermediate and high risk patients with NMIBC who underwent Bacillus Calmette-Guerin (BCG) induction therapy or delayed intravesical chemotherapy between 1999 and 2014. We introduced SI of 30 mg THP in 30 ml normal saline for all cases in 2010, and thus earlier cases could be considered as historical controls. As BCG induction therapy, patients received 80 mg of BCG Tokyo strain 2 weeks after transurethral resection of bladder tumor (TURBT), and the instillations were repeated weekly for 8 weeks. On the other hand, as delayed intravesical chemotherapy, patients received 30 mg THP in 30 ml normal saline over a period of 6 months starting 2 weeks after TURBT. The instillation schedule was once a week for 1 month, every other week for 1 month and once a month for 4 months. The patients were followed with cystoscopy and urine cytology every 3 months for the first 2 years and every 6 months thereafter. The 3-, and 5-year non-recurrence rates were 80. 3 and 80.3%, respectively, in the single immediate instillation group and 69.7 and 64.5%, respectively, in the control group. Univariate analysis revealed a significant difference between the SI group and the control group (P=0. 025). Multivariate analysis showed that there was an independent and significant recurrence risk factor in selecting chemotherapy instead of BCG in additional intravesical instillation therapy and not to perform SI. Limitations of our study are its retrospective and nonrandomized nature with a limited number of patients.

Safety and efficacy of various strains of bacille Calmette–Guérin in the treatment of bladder tumors in standard clinical practice

BackgroundThe natural progression of bladder tumors (nonmuscle-invasive bladder cancer [NMIBC]) is recurrence with a high rate of progression. Bacille Calmette–Guérin (BCG) has been shown effective in reducing these rates, but there are few comparative studies between strains. Material and methodsAn observational, prospective and multicentre registry studied 433 patients with a 12-month follow-up visit from 961 registered patients, assessing disease-free survival (DFS), progression-free survival (PFS) cancer-specific survival (CSS) and adverse effects. We studied the Tice, Russian, Tokyo, Connaught and RIVM strains. ResultsThe sociodemographic data, NMIBC history, comorbidities, size, number, stage, grade, associated carcinoma in situ and transurethral resection were well balanced. DFSThere were 85 relapses (19.6%). The median DFS time was 20months. When comparing the various strains, we detected no statistically significant differences (log-rank test; p=0.93). LPS: There were 33 cases of progression (7.62%). When comparing the various strains, we detected no statistically significant differences (log-rank test; p=0.69). CSS: Seven patients died (1.68%). When comparing the various strains, we detected no statistically significant differences (log-rank test; p=0.93).In terms of safety, 33.3% of the patients presented some type of adverse effect, mostly lower urinary symptoms (no urinary tract infections) <48h, >48h and haematuria. According to the Common Toxicity Criteria of the European Organisation for Research and Treatment of Cancer, 92.7% of the patients were grade 1. There were no statistically significant differences between the strains. ConclusionsIn this intermediate analysis, the risk of recurrence, progression, specific death and safety were independent of the BCG strain employed.

Eficacia y seguridad de distintas cepas de BCG en el tratamiento del tumor vesical en práctica clínica habitual

BackgroundThe natural progression of bladder tumours (nonmuscle-invasive bladder cancer [NMIBC]) is recurrence with a high rate of progression. Bacille Calmette-Guérin (BCG) has been shown effective in reducing these rates, but there are few comparative studies between strains. Material and methodsAn observational, prospective and multicentre registry studied 433 patients with a 12-month follow-up visit from 961 registered patients, assessing disease-free survival (DFS), progression-free survival (PFS) cancer-specific survival (CSS) and adverse effects. We studied the Tice, Russian, Tokyo, Connaught and RIVM strains. ResultsThe sociodemographic data, NMIBC history, comorbidities, size, number, stage, grade, associated carcinoma in situ and transurethral resection were well balanced.DFS: There were 85 relapses (19.6%). The median DFS time was 20months. When comparing the various strains, we detected no statistically significant differences (log-rank test; P=.93). LPS: There were 33 cases of progression (7.62%). When comparing the various strains, we detected no statistically significant differences (log-rank test; P=.69). CSS: Seven patients died (1.68%). When comparing the various strains, we detected no statistically significant differences (log-rank test; P=.93).In terms of safety, 33.3% of the patients presented some type of adverse effect, mostly lower urinary symptoms (no urinary tract infections) <48h, >48h and haematuria. According to the Common Toxicity Criteria of the European Organisation for Research and Treatment of Cancer, 92.7% of the patients were grade1. There were no statistically significant differences between the strains. ConclusionsIn this intermediate analysis, the risk of recurrence, progression, specific death and safety were independent of the BCG strain employed.

Genetic characteristics of Mycobacterium bovis BCG Thai Red Cross Society strain

Background: Bacillus Calmette-Guerin (BCG) Tokyo 172-1 strain has been used as a parent strain since 1988 by Queen Saovabha Memorial Institute, Thai Red Cross Society to manufacture BCG vaccine in Thailand. In general,the BCG Tokyo strain includes 2 BCG type populations: types I and II. Several studies have found that the type I population is higher than type II. Polymerase chain reaction has shown the presence of 2 types ofsubpopulations in the BCG Thai Red Cross Society (TRCS) strain. However, the characteristics of all genes from BCG TRCS have not been studied in depth, including whether or not the original features of the BCG Tokyo 172-1 strain are still retained. Objectives: To compare BCG TRCS and BCG Tokyo 172-1 for differences between their genomes. Methods: The whole genome of BCG TRCS was analyzed by next-generation sequencing. Results: BCG TRCS contained 23 different points and was 4 bases less in length than BCG Tokyo 172-1. However, its main features remain similar to those of the original BCG Tokyo 172-1 strain. Conclusions: The changes in the genome may be caused by a variety of factors including the strain of Tokyo 172, vaccine processing, storage, and natural changes in the genome. Keywords: BCG, BCG Tokyo, Mycobacterium bovis, next-generation sequencing

Bacteria Endosymbiont, Wolbachia, Promotes Parasitism of Parasitoid Wasp Asobara japonica.

Wolbachia is the most widespread endosymbiotic bacterium that manipulates reproduction of its arthropod hosts to enhance its own spread throughout host populations. Infection with Wolbachia causes complete parthenogenetic reproduction in many Hymenoptera, producing only female offspring. The mechanism of such reproductive manipulation by Wolbachia has been extensively studied. However, the effects of Wolbachia symbiosis on behavioral traits of the hosts are scarcely investigated. The parasitoid wasp Asobara japonica is an ideal insect to investigate this because symbiotic and aposymbiotic strains are available: Wolbachia-infected Tokyo (TK) and noninfected Iriomote (IR) strains originally collected on the main island and southwest islands of Japan, respectively. We compared the oviposition behaviors of the two strains and found that TK strain females parasitized Drosophila melanogaster larvae more actively than the IR strain, especially during the first two days after eclosion. Removing Wolbachia from the TK strain wasps by treatment with tetracycline or rifampicin decreased their parasitism activity to the level of the IR strain. Morphological and behavioral analyses of both strain wasps showed that Wolbachia endosymbionts do not affect development of the host female reproductive tract and eggs, but do enhance host-searching ability of female wasps. These results suggest the possibility that Wolbachia endosymbionts may promote their diffusion and persistence in the host A. japonica population not only at least partly by parthenogenesis but also by enhancement of oviposition frequency of the host females.

J0440203 Ti-Ni超弾性合金のカメラ保持用補助具への応用

The purpose of this study is to application of camera posture aid using Ti-Ni superelasticitc alloy. The composition of this alloy is Ti-50.8at%Ni. This alloy showed the superelastic behavior at room temperature. The maximum load of superelastic wire is 11N by three point of bending test. The superelastic component of camera posture aid was made from sixteen Ti-Ni wires of 1.0mm in diameter and 170mm in length. The camera posture aid has a part and the part of the prop to attach to an arm. The component arranged on the arm part and prop part. The arm part was made with 3D printer. The weight of camera posture aid is 200g. Mechanical property of the component was using a load cell, Tokyo Sokki kenkyujo TCLZ-5KNA and strain gage, KYOWA ELECTRONIC INSTRUMANTS. The maximum load of the camera posture aid is 7N. The camera shake was reduced about 65.5% by camera posture aid.

Development of a BCG challenge model for the testing of vaccine candidates against tuberculosis in cattle

Vaccination is being considered as part of a sustainable strategy for the control of bovine tuberculosis (BTB) in the UK. The live attenuated Mycobacterium bovis bacillus Calmette-Guerin (BCG) has been used experimentally to vaccinate cattle against BTB. However, BCG confers partial protection against BTB and therefore, there is a need to develop improved vaccines. BTB vaccine efficacy experiments require the use of biosafety level 3 facilities which are expensive to maintain, generally oversubscribed and represent a bottle neck for the testing of vaccine candidates. One indicator of the induction of protective responses would be the ability of the host's immune response to control/kill mycobacteria. In this work we have evaluated an intranodal BCG challenge for the selection of vaccine candidates at biosafety level 2 which are capable of inducing mycobactericidal responses. To our knowledge, this is the first such report. Whilst BCG only confers partial protection, it is still the standard against which other vaccines are judged. Therefore we tested the BCG intranodal challenge in BCG (Danish strain) vaccinated cattle and showed that vaccinated cattle had lower BCG cfu counts than naïve cattle at 14 and 21 days after intranodal challenge with BCG (Tokyo strain). This model could help prioritize competing TB vaccine candidates and exploration of primary and secondary immune responses to mycobacteria.

Genome sequencing and analysis of BCG vaccine strains.

BackgroundAlthough the Bacillus Calmette-Guérin (BCG) vaccine against tuberculosis (TB) has been available for more than 75 years, one third of the world's population is still infected with Mycobacterium tuberculosis and approximately 2 million people die of TB every year. To reduce this immense TB burden, a clearer understanding of the functional genes underlying the action of BCG and the development of new vaccines are urgently needed.Methods and FindingsComparative genomic analysis of 19 M. tuberculosis complex strains showed that BCG strains underwent repeated human manipulation, had higher region of deletion rates than those of natural M. tuberculosis strains, and lost several essential components such as T-cell epitopes. A total of 188 BCG strain T-cell epitopes were lost to various degrees. The non-virulent BCG Tokyo strain, which has the largest number of T-cell epitopes (359), lost 124. Here we propose that BCG strain protection variability results from different epitopes. This study is the first to present BCG as a model organism for genetics research. BCG strains have a very well-documented history and now detailed genome information. Genome comparison revealed the selection process of BCG strains under human manipulation (1908–1966).ConclusionsOur results revealed the cause of BCG vaccine strain protection variability at the genome level and supported the hypothesis that the restoration of lost BCG Tokyo epitopes is a useful future vaccine development strategy. Furthermore, these detailed BCG vaccine genome investigation results will be useful in microbial genetics, microbial engineering and other research fields.