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Veterinary Immunology and... Jul 2024Bovine tuberculosis (bTB) represents a threat to livestock production. Mycobacterium bovis is the main causative agent of bTB and a pathogen capable of infecting...
Bovine tuberculosis (bTB) represents a threat to livestock production. Mycobacterium bovis is the main causative agent of bTB and a pathogen capable of infecting wildlife and humans. Eradication programs based on surveillance in slaughterhouses with mandatory testing and culling of reactive cattle have failed to eradicate bTB in many regions worldwide. Therefore, developing effective tools to control this disease is crucial. Using a computational tool, we identified proteins in the M. bovis proteome that carry predictive binding peptides to BoLADRB3.2 and selected Mb0309, Mb1090, Mb1810 and Mb3810 from all the identified proteins. The expression of these proteins in a baculovirus-insect cell expression system was successful only for Mb0309 and Mb3810. In parallel, we expressed the ESAT-6 family proteins EsxG and EsxH in this system. Among the recombinant proteins, Mb0309 and EsxG exhibited moderate performance in distinguishing between cattle that test positive and negative to bTB using the official test, the intradermal tuberculin test (IDT), when used to stimulate interferon-gamma production in blood samples from cattle. However, when combined as a protein cocktail, Mb0309 and EsxG were reactive in 50 % of positive cattle. Further assessments in cattle that evade the IDT (false negative) and cattle infected with Mycobacterium avium paratuberculosis are necessary to determine the potential utility of this cocktail as an additional tool to assist the accurate diagnosis of bTB.
Topics: Mycobacterium bovis; Animals; Cattle; Antigens, Bacterial; Tuberculosis, Bovine; Bacterial Proteins; Tuberculin Test; Recombinant Proteins
PubMed: 38838485
DOI: 10.1016/j.vetimm.2024.110788 -
Frontiers in Veterinary Science 2024Ante-mortem diagnosis of bovine tuberculosis (bTB) is based mainly on the tuberculin skin test (TST) and the ɣ-IFN release assay (IGRA). Some infected animals escape...
Ante-mortem diagnosis of bovine tuberculosis (bTB) is based mainly on the tuberculin skin test (TST) and the ɣ-IFN release assay (IGRA). Some infected animals escape screening tests, thus, limit herd sanitation. Previous reports have suggested a predominant pattern of multi-organ lesions attributable to (the causative agent of bTB) bacteraemia. A case-control study was conducted to investigate blood PCR as an alternative tool for improving ante-mortem detection of TST false-negative bovines. Cases comprised 70 TST false-negative bovines (cases), which were serology positive, and controls included 81 TST positive bovines; all of them confirmed as infected with . Detection of the IS target through touchdown blood-PCR (IS TD-PCR) was performed. The positivity of the blood-PCR was 27.2% in the control group. This performance was similar to the 15% obtained among cases ( = 0.134). Most cases identified by the IS TD-PCR exhibited focalized lesions ( = 0.002). Results demonstrated that blood-PCR could detect TST false-negative cattle, even if they are negative for IGRA. Considering that cases exhibited humoral response to , further studies conducted in a pre-serological stage could provide evidence about the real contribution of the technique in herds.
PubMed: 38835898
DOI: 10.3389/fvets.2024.1359205 -
Frontiers in Immunology 2024Bacillus Calmette-Guérin (BCG) is the first line treatment for bladder cancer and it is also proposed for melanoma immunotherapy. BCG modulates the tumor...
Bacillus Calmette-Guérin (BCG) is the first line treatment for bladder cancer and it is also proposed for melanoma immunotherapy. BCG modulates the tumor microenvironment (TME) inducing an antitumor effective response, but the immune mechanisms involved still poorly understood. The immune profile of B16-F10 murine melanoma cells was assessed by infecting these cells with BCG or stimulating them with agonists for different innate immune pathways such as TLRs, inflammasome, cGAS-STING and type I IFN. B16-F10 did not respond to any of those stimuli, except for type I IFN agonists, contrasting with bone marrow-derived macrophages (BMDMs) that showed high production of proinflammatory cytokines. Additionally, we confirmed that BCG is able to infect B16-F10, which in turn can activate macrophages and spleen cells from mice in co-culture experiments. Furthermore, we established a subcutaneous B16-F10 melanoma model for intratumoral BCG treatment and compared wild type mice to TLR2, TLR3, TLR4, TLR7, TLR3/7/9, caspase 1, caspase 11, IL-1R, cGAS, STING, IFNAR, MyD88deficient animals. These results demonstrate that MyD88 signaling is important for BCG immunotherapy to control melanoma in mice. Also, BCG fails to induce cytokine production in the co-culture experiments using B16-F10 and BMDMs or spleen cells derived from MyD88 compared to wild-type (WT) animals. Immunotherapy with BCG was not able to induce the recruitment of inflammatory cells in the TME from MyD88 mice, impairing tumor control and IFN-γ production by T cells. In conclusion, MyD88 impacts on both innate and adaptive responses to BCG leading to an efficient antitumor response against melanoma.
Topics: Animals; Myeloid Differentiation Factor 88; Melanoma, Experimental; Mice; Signal Transduction; BCG Vaccine; Immunotherapy; Mice, Knockout; Mice, Inbred C57BL; Tumor Microenvironment; Cell Line, Tumor; Macrophages; Mycobacterium bovis; Cytokines
PubMed: 38835781
DOI: 10.3389/fimmu.2024.1380069 -
Journal of Vascular Surgery Cases and... Aug 2024Bacillus Calmette-Guérin (BCG), a live attenuated form of is part of the therapy for non-muscle-invasive bladder cancer. Cases of vascular graft infection in the...
Bacillus Calmette-Guérin (BCG), a live attenuated form of is part of the therapy for non-muscle-invasive bladder cancer. Cases of vascular graft infection in the context of BCG dissemination are rarely reported in the literature. We report a case of a 77-year-old man, who underwent intravesical instillation of BCG approximately 10 years earlier and presented to our hospital with acute thrombosis of a previous aortobisiliac graft, which tested positive for BCG infection. Aortic graft infections due to BCG dissemination are rare, but possible, complications. A prompt and multidisciplinary approach is necessary.
PubMed: 38818509
DOI: 10.1016/j.jvscit.2024.101504 -
PLoS Neglected Tropical Diseases May 2024Bovine tuberculosis (bTB) is a chronic zoonotic disease affecting cattle of all age groups including wild animals. It poses a significant threat to public health and...
Bovine tuberculosis (bTB) is a chronic zoonotic disease affecting cattle of all age groups including wild animals. It poses a significant threat to public health and high economic losses to dairy farmers. While the disease has been eradicated from most of the developed countries through extensive surveillance, testing and culling strategy, it is endemic in Africa, Asia, and the Middle East countries. Currently, there is limited research regarding the prevalence of bTB in cattle in Bhutan. This study aimed to determine the seroprevalence of bTB in cattle in six districts of eastern Bhutan. A two-stage probability proportional to size (PPS) sampling strategy was used to determine the number of animals from which serum samples needed to be collected in each district and sub-district. All farms and cattle for sampling were randomly selected from the data in the annual livestock census of 2020. The samples were tested using bTB ELISA test kit. The seroprevalence and their 95% confidence intervals were calculated. Logistic regression models were constructed to assess the influence of various individual animal and environmental risk factors (breed, age, sex, source of animal, body condition scores of animals, respiratory system status) associated with sero-positivity in animals. The study revealed an apparent seroprevalence of 2.57% (25/971 cattle; 95% CI:1.58-3.57), with an estimated true seroprevalence of 0.91% (95% CI: 0.0-2.81). However, none of the variables were found to be significantly associated with bTB seroprevalence in cattle. We recommend, further sampling and employment of confirmatory testing to fully ascertain the extent of bTB in the cattle herds in eastern Bhutan for prevention and control.
Topics: Animals; Cattle; Bhutan; Seroepidemiologic Studies; Tuberculosis, Bovine; Risk Factors; Female; Male; Mycobacterium bovis; Prevalence; Antibodies, Bacterial
PubMed: 38805568
DOI: 10.1371/journal.pntd.0012223 -
Frontiers in Immunology 2024Mycobacteria are known to exert a range of heterologous effects on the immune system. The mycobacteria-based Freund's Complete Adjuvant is a potent non-specific...
INTRODUCTION
Mycobacteria are known to exert a range of heterologous effects on the immune system. The mycobacteria-based Freund's Complete Adjuvant is a potent non-specific stimulator of the immune response used in immunization protocols promoting antibody production, and Mycobacterium bovis Bacille Calmette Guérin (BCG) vaccination has been linked with decreased morbidity and mortality beyond the specific protection it provides against tuberculosis (TB) in some populations and age groups. The role of heterologous antibodies in this phenomenon, if any, remains unclear and under-studied.
METHODS
We set out to evaluate antibody responses to a range of unrelated pathogens following infection with Mycobacterium tuberculosis (M.tb) and vaccination with BCG or a candidate TB vaccine, MTBVAC, in non-human primates.
RESULTS
We demonstrate a significant increase in the titer of antibodies against SARS-CoV-2, cytomegalovirus, Epstein-Barr virus, tetanus toxoid, and respiratory syncytial virus antigens following low-dose aerosol infection with M.tb. The magnitude of some of these responses correlated with TB disease severity. However, vaccination with BCG administered by the intradermal, intravenous or aerosol routes, or intradermal delivery of MTBVAC, did not increase antibody responses against unrelated pathogens.
DISCUSSION
Our findings suggest that it is unlikely that heterologous antibodies contribute to the non-specific effects of these vaccines. The apparent dysregulation of B cell responses associated with TB disease warrants further investigation, with potential implications for risk of B cell cancers and novel therapeutic strategies.
Topics: Animals; BCG Vaccine; Tuberculosis; Mycobacterium tuberculosis; Vaccination; Antibodies, Bacterial; Antibodies, Viral; Tuberculosis Vaccines; Female; Macaca mulatta; SARS-CoV-2; COVID-19; Immunity, Heterologous; Male
PubMed: 38799468
DOI: 10.3389/fimmu.2024.1387454 -
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi =... May 2024Objective To explore the regulatory role of dual-specificity phosphatase 5 (DUSP5) in BCG-mediated inflammatory response in mouse RAW264.7 macrophages. Methods Western...
Objective To explore the regulatory role of dual-specificity phosphatase 5 (DUSP5) in BCG-mediated inflammatory response in mouse RAW264.7 macrophages. Methods Western blot analysis was employed to detect the expression changes of DUSP5 in BCG-infected RAW264.7 macrophages at the period of 0.5, 1, 2, 4, 6, 8, 12 and 24 hours. Intracellular DUSP5 was reduced by small interfering RNA (siRNA) and transfected RAW264.7 macrophages were divided into siRNA-negative control (si-NC) group, DUSP5 knockdown (si-DUSP5) group, si-NC combined BCG infection group, and si-DUSP5 combined BCG infection group. Real-time quantitative PCR was conducted to measure the mRNA expression of interleukin 1β (IL-1β), IL-6, tumor necrosis factor α (TNF-α), and IL-10 in cells. ELISA was performed to measure the concentration of the cytokines in cell culture medium. Western blot analysis was performed to detect the expression changes of cellular nuclear factor κB (NF-κB) and phosphorylated NF-κB (p-NF-κB). Results BCG infection upregulated DUSP5 protein expression in RAW264.7 macrophages with the expression of DUSP5 reaching the peak after 4 hours' BCG stimulation. Comparing with si-NC combined BCG infection group, DUSP5 knockdown inhibited the expression and secretion of pro-inflammatory factors IL-1β, IL-6, and TNF-α, while the expression of the anti-inflammatory factor IL-10 was not affected by DUSP5. Moreover, knockdown of DUSP5 inhibited the phosphorylation of NF-κB in cells. Conclusion DUSP5 knockdown inhibites BCG-mediated macrophage inflammatory response via blocking NF-κB signaling activation.
Topics: Animals; Mice; RAW 264.7 Cells; Dual-Specificity Phosphatases; NF-kappa B; Signal Transduction; Macrophages; Inflammation; Gene Knockdown Techniques; Mycobacterium bovis; Cytokines
PubMed: 38790096
DOI: No ID Found -
Scientific Reports May 2024We have previously reported the transcriptomic and lipidomic profile of the first-generation, hygromycin-resistant (Hyg) version of the BCGΔBCG1419c vaccine candidate,... (Comparative Study)
Comparative Study
Comparison of the transcriptome, lipidome, and c-di-GMP production between BCGΔBCG1419c and BCG, with Mincle- and Myd88-dependent induction of proinflammatory cytokines in murine macrophages.
We have previously reported the transcriptomic and lipidomic profile of the first-generation, hygromycin-resistant (Hyg) version of the BCGΔBCG1419c vaccine candidate, under biofilm conditions. We recently constructed and characterized the efficacy, safety, whole genome sequence, and proteomic profile of a second-generation version of BCGΔBCG1419c, a strain lacking the BCG1419c gene and devoid of antibiotic markers. Here, we compared the antibiotic-less BCGΔBCG1419c with BCG. We assessed their colonial and ultrastructural morphology, biofilm, c-di-GMP production in vitro, as well as their transcriptomic and lipidomic profiles, including their capacity to activate macrophages via Mincle and Myd88. Our results show that BCGΔBCG1419c colonial and ultrastructural morphology, c-di-GMP, and biofilm production differed from parental BCG, whereas we found no significant changes in its lipidomic profile either in biofilm or planktonic growth conditions. Transcriptomic profiling suggests changes in BCGΔBCG1419c cell wall and showed reduced transcription of some members of the DosR, MtrA, and ArgR regulons. Finally, induction of TNF-α, IL-6 or G-CSF by bone-marrow derived macrophages infected with either BCGΔBCG1419c or BCG required Mincle and Myd88. Our results confirm that some differences already found to occur in Hyg BCGΔBCG1419c compared with BCG are maintained in the antibiotic-less version of this vaccine candidate except changes in production of PDIM. Comparison with previous characterizations conducted by OMICs show that some differences observed in BCGΔBCG1419c compared with BCG are maintained whereas others are dependent on the growth condition employed to culture them.
Topics: Animals; Myeloid Differentiation Factor 88; Mice; Macrophages; Transcriptome; Lipidomics; BCG Vaccine; Cyclic GMP; Mycobacterium bovis; Biofilms; Cytokines; Membrane Proteins; Gene Expression Profiling; Lectins, C-Type
PubMed: 38789479
DOI: 10.1038/s41598-024-61815-8 -
Journal of Parkinson's Disease May 2024Deep brain stimulation (DBS) is an advanced treatment in Parkinson's disease. We describe a 71-year-old patient in whom the DBS got infected with Mycobacterium bovis...
Deep brain stimulation (DBS) is an advanced treatment in Parkinson's disease. We describe a 71-year-old patient in whom the DBS got infected with Mycobacterium bovis shortly after intravesical BCG instillations as an adjuvant treatment of bladder cancer. The DBS internal pulse generator and extension wires had to be replaced, and the patient was treated successfully with rifampicin, isoniazid, and ethambutol during three months. This case suggests that physicians need to be aware of the risk of this kind of infection and add a specific Mycobacterial test to the regular cultures.
PubMed: 38788088
DOI: 10.3233/JPD-230426 -
Veterinary Sciences May 2024Bovine tuberculosis is a zoonotic disease of significant impact, particularly in countries where a pastoral economy is predominant. Despite its importance, few studies...
INTRODUCTION
Bovine tuberculosis is a zoonotic disease of significant impact, particularly in countries where a pastoral economy is predominant. Despite its importance, few studies have analysed the disease's behaviour in Colombia, and none have developed maps using geographic information systems (GIS) to characterise it; as such, we developed this study to describe the temporal-spatial distribution of bovine tuberculosis in Colombia over a period of 19 years.
METHODS
A retrospective cross-sectional descriptive study, based on reports by the Colombian Agricultural Institute (ICA), surveillance of tuberculosis on cattle farms in Colombia from 2001 to 2019 was carried out. The data were converted into databases using Microsoft Access 365, and multiple epidemiological maps were generated with the QGIS version 3.36 software coupled to shape files of all the country's departments.
RESULTS
During the study period, 5273 bovine tuberculosis cases were identified in multiple different departments of Colombia (with a mean of 278 cases/year). Regarding its temporal distribution, the number of cases varied from a maximum of 903 cases (17.12% of the total) in 2015 to a minimum of 0 between 2001 and 2004 and between 2017 and 2019 (between 2005 and 2016, the minimum was 46 cases, 0.87%).
CONCLUSIONS
GIS are essential for understanding the temporospatial behaviour of zoonotic diseases in Colombia, as is the case for bovine tuberculosis, with its potential implications for the Human and One Health approaches.
PubMed: 38787192
DOI: 10.3390/vetsci11050220