Subcutaneous Administration of a 10-Fold-Lower Dose of a Commercial Human Tuberculosis Vaccine, Mycobacterium bovis Bacillus Calmette-Guérin Danish, Induced Levels of Protection against Bovine Tuberculosis and Responses in the Tuberculin Intradermal Test Similar to Those Induced by a Standard Cattle Dose

ABSTRACTVaccination of cattle with a commercial human tuberculosis (TB) vaccine,Mycobacterium bovisbacillus Calmette-Guérin (BCG) Danish, at a dose equivalent to 5 human doses of BCG has protected these animals against TB in field and experimental trials. There is interest in determining whether a 10-fold-lower dose could still protect cattle but not induce a tuberculin intradermal test response. Two groups of calves (n= 9/group) were vaccinated subcutaneously with a lyophilized BCG Danish vaccine containing either 0.5 (1 × 105to 4 × 105CFU) or 5 (1 × 106to 4 × 106CFU) human doses of BCG Danish, with an additional group of 10 calves serving as nonvaccinated controls. Fifteen weeks after vaccination, these animals were challenged intratracheally with 5 × 103CFU of virulentM. bovisand another 15 weeks later were slaughtered and examined for the presence of tuberculous lesions. Vaccination of the calves with either 0.5 or 5 equivalent human doses of BCG Danish induced similar levels of protection against challenge withM. bovis, with both groups showing significant reductions in the pathological and microbiological parameters compared to those for the the control group (P< 0.05). Vaccination with either of the two BCG doses induced similar numbers of animals responding to the tuberculin intradermal test at 11 weeks postvaccination. Vaccination with a 0.5 equivalent human dose of a commercial lyophilized BCG vaccine can protect cattle against challenge withM. bovis.

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Improved Skin Test for Differential Diagnosis of Bovine Tuberculosis by the Addition of Rv3020c-Derived Peptides

Clinical and Vaccine Immunology ◽

10.1128/cvi.00024-12 ◽

2012 ◽

Vol 19 (4) ◽

pp. 620-622 ◽

Cited By ~ 33

Author(s):

Gareth J. Jones ◽

Adam Whelan ◽

Derek Clifford ◽

Mick Coad ◽

H. Martin Vordermeier

Keyword(s):

Differential Diagnosis ◽

Skin Test ◽

Mycobacterium Bovis ◽

Bovine Tuberculosis ◽

Johne's Disease ◽

Diagnostic Sensitivity ◽

Bacillus Calmette Guerin ◽

Content Type ◽

The Face ◽

Mycobacterial Antigens

ABSTRACTA peptide cocktail derived from the mycobacterial antigens ESAT-6, CFP-10, and Rv3615c allowed differentiation betweenMycobacterium bovis-infected and M. bovis bacillus Calmette-Guérin (BCG)-vaccinated cattle when used as a skin test reagent for a “DIVA” test (i.e., a test capable ofdifferentiatinginfected and uninfectedvaccinatedanimals). Addition of the antigen Rv3020c improves the diagnostic sensitivity without compromising specificity in the face of BCG or Johne's disease vaccination.

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Tonsils of the Soft Palate Do Not Mediate the Response of Pigs to Oral Vaccination with Heat-Inactivated Mycobacterium bovis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00221-14 ◽

2014 ◽

Vol 21 (8) ◽

pp. 1128-1136 ◽

Cited By ~ 8

Author(s):

Beatriz Beltrán-Beck ◽

Beatriz Romero ◽

Mariana Boadella ◽

Carmen Casal ◽

Javier Bezos ◽

...

Keyword(s):

Wild Boar ◽

Mycobacterium Bovis ◽

Mononuclear Cells ◽

Mammalian Species ◽

Serum Antibody ◽

Control Group ◽

Mrna Levels ◽

Oral Vaccination ◽

Peripheral Blood Mononuclear ◽

Content Type

ABSTRACTMycobacterium boviscauses animal tuberculosis (TB) in cattle, humans, and other mammalian species, including pigs. The goal of this study was to experimentally assess the responses of pigs with and without a history of tonsillectomy to oral vaccination with heat-inactivatedM. bovisand challenge with a virulentM. bovisfield strain, to compare pig and wild boar responses using the same vaccination model as previously used in the Eurasian wild boar (Sus scrofa), to evaluate the use of several enzyme-linked immunosorbent assays (ELISAs) and lateral flow tests forin vivoTB diagnosis in pigs, and to verify if these tests are influenced by oral vaccination with inactivatedM. bovis. At necropsy, the lesion and culture scores were 20% to 43% higher in the controls than those in the vaccinated pigs. MassiveM. bovisgrowth from thoracic tissue samples was observed in 4 out of 9 controls but in none of the 10 vaccinated pigs. No effect of the presence or absence of tonsils was observed on these scores, suggesting that tonsils are not involved in the protective response to this vaccine in pigs. The serum antibody levels increased significantly only after challenge. At necropsy, the estimated sensitivities of the ELISAs and dual path platform (DPP) assays ranged from 89% to 94%. In the oral mucosa, no differences in gene expression were observed in the control group between the pigs with and without tonsils. In the vaccinated group, the mRNA levels for chemokine (C-C motif) receptor 7 (CCR7), interferon beta (IFN-β), and methylmalonyl coenzyme A mutase (MUT) were higher in pigs with tonsils. Complement component 3 mRNA levels in peripheral blood mononuclear cells (PBMC) increased with vaccination and decreased afterM. bovischallenge. This information is relevant for pig production in regions that are endemic forM. bovisand for TB vaccine research.

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Interleukin-21 Induces Short-Lived Effector CD8+ T Cells but Does Not Inhibit Their Exhaustion after Mycobacterium bovis BCG Infection in Mice

Infection and Immunity ◽

10.1128/iai.00147-18 ◽

2018 ◽

Vol 86 (8) ◽

Cited By ~ 1

Author(s):

Naoto Noguchi ◽

Risa Nakamura ◽

Shinya Hatano ◽

Hisakata Yamada ◽

Xun Sun ◽

...

Keyword(s):

T Cells ◽

Mycobacterium Bovis ◽

Cd8 T Cells ◽

Absolute Number ◽

Bacillus Calmette Guerin ◽

Content Type ◽

Effector Functions ◽

Antigen 85B ◽

Interleukin 21 ◽

Recombinant Bcg

ABSTRACT Interleukin 21 (IL-21) is a pleiotropic common cytokine receptor γ chain cytokine that promotes the effector functions of NK cells and CD8+ T cells and inhibits CD8+ T cell exhaustion during chronic infection. We found that the absolute number of short-lived effector CD8+ T cells (SLECs) (KLRG1high CD127low) decreased significantly in IL-21 receptor-deficient (IL-21R−/−) mice during Mycobacterium bovis bacillus Calmette-Guérin (BCG) infection. Early effector CD8+ T cells (EECs) (KLRG1low CD127low) were normally generated in IL-21R−/− mice after infection. Exhausted CD8+ T cells (PD-1high KLRG1low) were also normally generated in IL-21R−/− mice after infection. Mixed bone marrow (BM) chimera and transfer experiments showed that IL-21R on CD8+ T cells was essential for the proliferation of EECs, allowing them to differentiate into SLECs after BCG infection. On the other hand, the number of SLECs increased significantly after infection with recombinant BCG (rBCG) that secreted an antigen 85B (Ag85B)–IL-21 fusion protein (rBCG–Ag85B–IL-21), but the number of exhausted CD8+ T cells did not change after rBCG–Ag85B–IL-21 infection. These results suggest that IL-21 signaling drives the differentiation of SLECs from EECs but does not inhibit the exhaustion of CD8+ T cells following BCG infection in mice.

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Duration of Immunity against Mycobacterium bovis following Neonatal Vaccination with Bacillus Calmette-Guérin Danish: Significant Protection against Infection at 12, but Not 24, Months

Clinical and Vaccine Immunology ◽

10.1128/cvi.00301-12 ◽

2012 ◽

Vol 19 (8) ◽

pp. 1254-1260 ◽

Cited By ~ 36

Author(s):

M. L. Thom ◽

M. McAulay ◽

H. M. Vordermeier ◽

D. Clifford ◽

R. G. Hewinson ◽

...

Keyword(s):

Lymph Node ◽

Immune Responses ◽

Mycobacterium Bovis ◽

Statistical Significance ◽

Significant Degree ◽

Bacillus Calmette Guerin ◽

Content Type ◽

Bcg Vaccination ◽

Ifn Γ ◽

Neonatal Calves

ABSTRACTVaccination of neonatal calves withMycobacterium bovisbacillus Calmette-Guérin (BCG) induces a significant degree of protection against bovine tuberculosis, caused by infection with virulentM. bovis. In two independent experiments, we assessed the duration of the protective immunity induced in calves by neonatal vaccination with BCG Danish. Protection from disease was assessed at 12 and 24 months postvaccination in cattle challenged via the endotracheal route withM. bovis. We also assessed antigen-specific immune responses to assess their utility as correlates of protection. At 12 months postvaccination, significant reductions in lung and lymph node pathologies were observed compared to nonvaccinatedM. bovis-challenged control cattle. At 24 months post-BCG vaccination, there was a reduction in lung and lymph node pathology scores and in bacterial burden. However, when comparing vaccinated and control groups, this did not reach statistical significance. Vaccination induced long-lived antigen (purified protein derivative [PPD])-specific gamma interferon (IFN-γ) release in whole-blood cultures, which remained above baseline levels for more than 20 months (approximately 90 weeks). The number of antigen-specific IFN-γ-secreting central memory T cells present at the time ofM. bovischallenge was significantly higher in vaccinated than in control animals at 12 months postvaccination, but not at 24 months. Vaccination of neonatal calves with BCG Danish induced protective immune responses against bovine TB which were maintained for at least 12 months postvaccination. These studies provide data on the immunity induced by BCG vaccination in calves; the results could inform vaccination strategies for the control of bovine TB in United Kingdom cattle herds.

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Protection by a Recombinant Mycobacterium bovis Bacillus Calmette-Guérin Vaccine Expressing Shiga Toxin 2 B Subunit against Shiga Toxin-Producing Escherichia coli in Mice

Clinical and Vaccine Immunology ◽

10.1128/cvi.00473-12 ◽

2012 ◽

Vol 19 (12) ◽

pp. 1932-1937 ◽

Cited By ~ 12

Author(s):

Jun Fujii ◽

Mariko Naito ◽

Takashi Yutsudo ◽

Sohkichi Matsumoto ◽

Daniel P. Heatherly ◽

...

Keyword(s):

Escherichia Coli ◽

Mycobacterium Bovis ◽

Shiga Toxin ◽

Bacillus Calmette Guerin ◽

Vaccine Strategy ◽

Content Type ◽

B Subunit ◽

Serum Igg ◽

Shiga Toxin 2 ◽

Potential Vaccine

ABSTRACTWe have developed a novel vaccine against Shiga toxin (Stx)-producingEscherichia coli(STEC) infection using a recombinantMycobacterium bovisBCG (rBCG) system. Two intraperitoneal vaccinations with rBCG expressing the Stx2 B subunit (Stx2B) resulted in an increase of protective serum IgG and mucosal IgA responses to Stx2B in BALB/c mice. When orally challenged with 103CFU of STEC strain B2F1 (O91: H21), the immunized mice survived statistically significantly longer than the nonvaccinated mice. We suggest that intraperitoneal immunization with rBCG expressing Stx2B would be a potential vaccine strategy for STEC.

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Ceftaroline versus Ceftriaxone in a Highly Penicillin-Resistant Pneumococcal Pneumonia Rabbit Model Using Simulated Human Dosing

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01773-09 ◽

2011 ◽

Vol 55 (7) ◽

pp. 3557-3563 ◽

Cited By ~ 24

Author(s):

Delphine Croisier-Bertin ◽

Lionel Piroth ◽

Pierre-Emmanuel Charles ◽

Aurélie Larribeau ◽

Donald Biek ◽

...

Keyword(s):

Bactericidal Activity ◽

Statistical Significance ◽

Rabbit Model ◽

Multidrug Resistant ◽

Control Group ◽

Bacterial Counts ◽

Content Type ◽

Human Dose ◽

Unit Reduction

ABSTRACTCeftaroline (CPT) is a new cephalosporin exhibiting bactericidal activity against Gram-positive organisms, including methicillin-resistantStaphylococcus aureus(MRSA) and multidrug-resistantStreptococcus pneumoniae(MDRSP), as well as common Gram-negative pathogens. This study investigated thein vivoefficacy of a 48-hour simulated human dose regimen of CPT compared with ceftriaxone (CRO) against isolates ofS. pneumoniaewith different susceptibilities to penicillin in a rabbit pneumonia model. ThreeS. pneumoniaestrains were used: CRO-susceptible penicillin-susceptibleS. pneumoniae(CRO-S PSSP), CRO-susceptible penicillin-intermediateS. pneumoniae(CRO-S PISP), and CRO-resistant penicillin-resistantS. pneumoniae(CRO-R PRSP). Animals were randomized to the control group (no treatment) (n= 22) or to a group given intravenous (IV) CPT human equivalent (HE) dosage (600 mg/12 h;n= 19) or IV CRO HE dosage (1 g/24 h;n= 19). The total doses needed to achieve the HE dosage were 71 and 82 mg/kg of body weight/24 h for CRO and CPT, respectively. One group of rabbits infected with the CRO-R PRSP strain received intramuscular (IM) administration of CPT (5 or 20 mg/kg twice daily;n= 5 for each). Evaluation of efficacy was based on bacterial counts in the lungs and spleen. For IV CPT and IV CRO, the mean areas under the concentration-time curves from 0 to 24 h (AUC0–24s) were 155 and 938 mg · h/liter, respectively, the maximum concentrations in serum (Cmaxs) were 20 and 158 mg/liter, respectively, and the minimum concentrations in serum (Cmins) were 1.3 and 6 mg/liter, respectively. Both agents effectively treated pulmonary infections caused by CRO-S PSSP or CRO-S PISP with complete bacterial eradication in the lungs and spleen after 2 days of treatment. Against PRSP, CPT demonstrated excellent bactericidal activity, reducing bacterial counts in the lungs and spleen by approximately 8 and 4 log units, respectively (P< 0.001); CRO treatment resulted in a 2-log-unit reduction in the bacterial counts in lungs that did not reach statistical significance. Twice-daily IM CPT (5 mg/kg) reduced the bacterial burden by approximately 6 log units in the lungs and 3 log units in the spleen, and the 20-mg/kg dosage effectively eradicated PRSP infection. These findings further validate thein vivobactericidal activity of CPT against pneumococci.

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Clofazimine Inhibits the Growth of Babesia and Theileria ParasitesIn VitroandIn Vivo

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01614-15 ◽

2016 ◽

Vol 60 (5) ◽

pp. 2739-2746 ◽

Cited By ~ 17

Author(s):

Bumduuren Tuvshintulga ◽

Mahmoud AbouLaila ◽

Batdorj Davaasuren ◽

Aki Ishiyama ◽

Thillaiampalam Sivakumar ◽

...

Keyword(s):

Subcutaneous Administration ◽

Blood Analysis ◽

Babesia Bovis ◽

Babesia Microti ◽

Control Group ◽

Blood Transfusions ◽

Inhibitory Effects ◽

Diminazene Aceturate ◽

Content Type

ABSTRACTThe present study evaluated the growth-inhibitory effects of clofazimine, currently used for treating leprosy, againstBabesia bovis,B. bigemina,B. caballi, andTheileria equiinin vitroculture and againstBabesia microtiin mice. The 50% inhibitory concentrations (IC50s) of clofazimine against thein vitrogrowth ofB. bovis,B. bigemina,B. caballi, andT. equiwere 4.5, 3, 4.3, and 0.29 μM, respectively. In mice infected withB. microti, treatment with 20 mg/kg of body weight of clofazimine administered orally resulted in a significantly lower peak parasitemia (5.3%) than that in the control group (45.9%), which was comparable to the subcutaneous administration of 25 mg/kg diminazene aceturate, the most widely used treatment for animal piroplasmosis. Although slight anemia was observed in both clofazimine- and diminazene aceturate-treated infected mice, the level and duration of anemia were lower and shorter, respectively, than those in untreated infected mice. Using blood transfusions and PCR, we also examined whether clofazimine completely killedB. microti. On day 40 postinfection, when blood analysis was performed, parasites were not found in blood smears; however, the DNA ofB. microtiwas detected in the blood of clofazimine-treated animals and in several tissues of clofazimine- and diminazene aceturate-treated mice by PCR. The growth of parasites was observed in mice after blood transfusions from clofazimine-treated mice. In conclusion, clofazimine showed excellent inhibitory effects againstBabesiaandTheileria in vitroandin vivo, and further study on clofazimine is required for the future development of a novel chemotherapy with high efficacy and safety against animal piroplasmosis and, possibly, human babesiosis.

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CD30 Is Required for Activation of a Unique Subset of Interleukin-17A-Producing γδ T Cells in Innate Immunity against Mycobacterium bovis Bacillus Calmette-Guérin Infection

Infection and Immunity ◽

10.1128/iai.00887-13 ◽

2013 ◽

Vol 81 (10) ◽

pp. 3923-3934 ◽

Cited By ~ 15

Author(s):

Ying Guo ◽

Xun Sun ◽

Kensuke Shibata ◽

Hisakata Yamada ◽

Hiromi Muta ◽

...

Keyword(s):

T Cells ◽

Mycobacterium Bovis ◽

Early Stage ◽

Γδ T Cells ◽

Bacillus Calmette Guerin ◽

Content Type ◽

Interleukin 17A ◽

Soluble Cd30 ◽

Infection Inhibition

ABSTRACTInterleukin-17A (IL-17A)-producing γδ T cells are known to be activated followingMycobacterium bovisbacillus Calmette-Guérin (BCG) infection. Here, we show that CD30, a member of the tumor necrosis factor (TNF) receptor superfamily, is important for activation of IL-17A-producing γδ T cells after BCG infection. Vγ1−Vγ4−γδ T cells preferentially expressing Vγ6/Vδ1 genes were identified as the major source of IL-17A in the peritoneal cavity during the early stage of BCG infection. The number of IL-17A-producing Vγ1−Vγ4−γδ T cells bearing Vγ6 increased in peritoneal exudate cells (PEC) of wild-type (WT) mice but not in those of CD30 knockout (KO) mice in response to BCG infection. Consistently, CD30 ligand (CD30L) or CD30 expression, predominantly by Vγ1−Vγ4−γδ T cells, was rapidly upregulated after BCG infection. Inhibition of CD30L/CD30 signaling byin vivoadministration of a soluble CD30 and immunoglobulin fusion protein (CD30-Ig) severely impaired activation of IL-17A-producing Vγ1−Vγ4−γδ T cells in WT mice, while stimulating CD30L/CD30 signaling byin vivoadministration of agonistic anti-CD30 monoclonal antibody (MAb) restored IL-17A production by Vγ1−Vγ4−γδ T cells in CD30L KO mice after BCG infection. These results suggest that CD30 signaling plays an important role in the activation of IL-17A-producing Vγ1−Vγ4−γδ T cells bearing Vγ6 at an early stage of BCG infection.

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Goats Primed with Mycobacterium bovis BCG and Boosted with a Recombinant Adenovirus Expressing Ag85A Show Enhanced Protection against Tuberculosis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00275-12 ◽

2012 ◽

Vol 19 (9) ◽

pp. 1339-1347 ◽

Cited By ~ 34

Author(s):

Bernat Pérez de Val ◽

Bernardo Villarreal-Ramos ◽

Miquel Nofrarías ◽

Sergio López-Soria ◽

Nadine Romera ◽

...

Keyword(s):

Mycobacterium Bovis ◽

Recombinant Adenovirus ◽

Bacterial Load ◽

Bacillus Calmette Guerin ◽

Content Type ◽

Vaccine Trials ◽

Natural Hosts ◽

Ifn Γ ◽

Cellular And Humoral Immunity ◽

Humoral Responses

ABSTRACTThis is the first efficacy study using the experimental goat model, a natural host of tuberculosis (TB), to evaluate the efficacy of heterologousMycobacterium bovisbacillus Calmette-Guérin (BCG) prime followed by boosting with a replication-deficient adenovirus expressing the antigen Ag85A (AdAg85A). Three experimental groups of 11 goat kids each were used: BCG vaccinated, BCG vaccinated and AdAg85A boosted, and nonvaccinated. Twenty-two goat kids were vaccinated with ∼5 × 105CFU of BCG (week 0), and 11 of them were boosted at week 8 with 109PFU of AdAg85A. At week 14, all goats were challenged by the endobronchial route with ∼1.5 × 103CFU ofMycobacterium caprae. The animals were euthanized at week 28. Cellular and humoral immunity induced by vaccination andM. capraeinfection was measured throughout the study. After challenge BCG-AdAg85A-vaccinated animals exhibited reduced pathology compared to BCG-vaccinated animals in lungs and in pulmonary lymph nodes. There were significant reductions in bacterial load in both groups of vaccinated goats, but the reduction was more pronounced in prime-boosted animals. Antigen-specific gamma interferon (IFN-γ) and humoral responses were identified as prognostic biomarkers of vaccination outcome depending on their correlation with pathological and bacteriological results. As far as we know, this is the first report using multidetector computed tomography (MDCT) to measure vaccine efficacy against pulmonary TB in an animal model. The use in vaccine trials of animals that are natural hosts of TB may improve research into human TB vaccines.

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Modulation effects of hexamethylene bisacetamide on growth and differentiation of cultured human malignant glioma cells

Journal of Neurosurgery ◽

10.3171/jns.1996.84.5.0831 ◽

1996 ◽

Vol 84 (5) ◽

pp. 831-838 ◽

Cited By ~ 13

Author(s):

Xiao-Nan Li ◽

Zi-Wei Du ◽

Qiang Huang

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Malignant Glioma ◽

Culture Media ◽

Morphological Changes ◽

Control Group ◽

Cell Cycle Distribution ◽

Content Type ◽

Hexamethylene Bisacetamide ◽

Human Malignant Glioma

✓ The modulation effects of hexamethylene bisacetamide (HMBA), a differentiation-inducing agent, on growth and differentiation of cells from human malignant glioma cell line SHG-44 were studied. At cytostatic doses (2.5 mM, 5 mM, 7.5 mM, and 10 mM for 15 days), HMBA exerted a marked inhibitory effect on cell proliferation. Exposure to HMBA (5 mM and 10 mM for 12 days) also resulted in an accumulation of cells in G0/G1 phase and a decrease of cells in S phase as analyzed by flow cytometry. The reversible effects of 7.5 mM HMBA and 10 mM HMBA on cell proliferation and 10 mM HMBA on disruption of cell cycle distribution were observed when HMBA was removed from culture media on Day 6 and replaced with HMBA-free media. Colony-forming efficiency (CFE) in soft agar was remarkably decreased by HMBA (2.5 mM, 5 mM, 7.5 mM, and 10 mM for 14 days), and in 7.5 mM HMBA— and 10 mM HMBA—treated cells, the CFEs were reduced to 25% and 12.5%, respectively, of that in untreated cells. Cells treated with HMBA (5 mM and 10 mM for 15 days) remained tumorigenic in athymic nude mice, but the growth rates of the xenografts were much slower than those in the control group. The effects of HMBA on cell proliferation, cell cycle distribution, CFE, and growth of xenografts were dose dependent. A more mature phenotype was confirmed by the morphological changes from spindle shape to large polygonal stellate shape and remarkably elevated expression of glial fibrillary acidic protein in cells exposed to HMBA (5 mM, 10 mM for 15 days). Our results showed that a more differentiated phenotype with marked growth arrest was induced in SHG-44 cells by HMBA.

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