TY - JOUR T1 - Allergic Asthma Favors Growth in the Lungs of Infected Mice. JF - Front Immunol Y1 - 2018 A1 - Arnaud Machelart A1 - Potemberg, Georges A1 - Laurye Van Maele A1 - Aurore Demars A1 - Maxime Lagneaux A1 - Carl De Trez A1 - Catherine Sabatel A1 - Bureau, Fabrice A1 - De Prins, Sofie A1 - Pauline Percier A1 - Olivier J Denis A1 - Fabienne Jurion A1 - Marta Romano A1 - Vanderwinden, Jean-Marie A1 - Letesson, Jean-Jacques A1 - Eric Muraille KW - allergic asthma KW - Brucella melitensis KW - Brucellosis KW - INFECTION KW - Mycobacterium tuberculosis KW - Streptococcus pneumoniae AB -

Allergic asthma is a chronic Th2 inflammatory disease of the lower airways affecting a growing number of people worldwide. The impact of infections and microbiota composition on allergic asthma has been investigated frequently. Until now, however, there have been few attempts to investigate the impact of asthma on the control of infectious microorganisms and the underlying mechanisms. In this work, we characterize the consequences of allergic asthma on intranasal (i.n.) infection by bacteria in mice. We observed that i.n. sensitization with extracts of the house dust mite or the mold () significantly increased the number of , and in the lungs of infected mice. Microscopic analysis showed dense aggregates of infected cells composed mainly of alveolar macrophages (CD11c F4/80 MHCII) surrounded by neutrophils (Ly-6G). Asthma-induced susceptibility appears to be dependent on CD4 T cells, the IL-4/STAT6 signaling pathway and IL-10, and is maintained in IL-12- and IFN-γR-deficient mice. The effects of the sensitization protocol were also tested on and pulmonary infections. Surprisingly, we observed that sensitization strongly increases the survival of infected mice by a T cell and STAT6 independent signaling pathway. In contrast, the course of infection is not affected in the lungs of sensitized mice. Our work demonstrates that the impact of the same allergic sensitization protocol can be neutral, negative, or positive with regard to the resistance of mice to bacterial infection, depending on the bacterial species.

VL - 9 M3 - 10.3389/fimmu.2018.01856 ER - TY - JOUR T1 - Helminth-induced IL-4 expands bystander memory CD8 T cells for early control of viral infection. JF - Nat Commun Y1 - 2018 A1 - Marion Rolot A1 - Annette M Dougall A1 - Alisha Chetty A1 - Javaux, Justine A1 - Chen, Ting A1 - Xiao, Xue A1 - Machiels, Bénédicte A1 - Murray E Selkirk A1 - Rick M Maizels A1 - Cornelis Hokke A1 - Olivier J Denis A1 - Frank Brombacher A1 - Vanderplasschen, Alain A1 - Gillet, Laurent A1 - William GC Horsnell A1 - Dewals, Benjamin G AB -

Infection with parasitic helminths can imprint the immune system to modulate bystander inflammatory processes. Bystander or virtual memory CD8 T cells (T) are non-conventional T cells displaying memory properties that can be generated through responsiveness to interleukin (IL)-4. However, it is not clear if helminth-induced type 2 immunity functionally affects the T compartment. Here, we show that helminths expand CD44CD62LCXCR3CD49d T cells through direct IL-4 signaling in CD8 T cells. Importantly, helminth-mediated conditioning of T cells provided enhanced control of acute respiratory infection with the murid gammaherpesvirus 4 (MuHV-4). This enhanced control of MuHV-4 infection could further be explained by an increase in antigen-specific CD8 T cell effector responses in the lung and was directly dependent on IL-4 signaling. These results demonstrate that IL-4 during helminth infection can non-specifically condition CD8 T cells, leading to a subsequently raised antigen-specific CD8 T cell activation that enhances control of viral infection.

VL - 9 CP - 1 M3 - 10.1038/s41467-018-06978-5 ER - TY - JOUR T1 - Relationship between mold exposure, specific IgE sensitization, and clinical asthma: A case-control study. JF - Ann Allergy Asthma Immunol Y1 - 2018 A1 - Muriel Vincent A1 - Francis Corazza A1 - Camille Chasseur A1 - Sandrine Bladt A1 - Marta Romano A1 - Huygen, Kris A1 - Olivier J Denis A1 - Olivier Michel AB -

BACKGROUND: Most of the findings related to the noxious effect of mold sensitization on asthma come from investigations based on Alternaria alternata, Cladosporium herbarum, and Aspergillus fumigatus. However, species such as Penicillium spp, Cladosporium sphaerospermum, Cladosporium cladosporioides, or Aspergillus versicolor display a more pronounced indoor tropism, and their potential harmful respiratory effects cannot be neglected.

OBJECTIVE: The goal of this work was to relate mold sensitizations with asthma severity and with the level of indoor mold contamination among mold-sensitized patients with asthma and nonsensitized patients with asthma.

METHODS: A case-control study was conducted and several asthma severity markers were compared between patients with asthma with and without mold sensitization. Indoor contamination of patients' dwellings was also investigated.

RESULTS: Our findings confirmed the association between sensitization to A fumigatus and severity for patients with asthma in contrast with sensitization to other species. Indoor mold contamination was detected in approximately 90% of dwellings. Overall mold exposure was not associated with asthma severity. However, regardless of the sensitization, exposure to A fumigatus and Penicillium spp in dust was linked to an increased risk of severe asthma.

CONCLUSION: The harmful nature of mold sensitization and mold exposure for patients with asthma was not confirmed in this study. However, sensitization to A fumigatus was associated with an increased risk for severe asthma. A better investigation of the properties of Penicillium spp is recommended because its exposure was found to be associated with a more pronounced impairment of lung function.

VL - 121 CP - 3 M3 - 10.1016/j.anai.2018.06.016 ER - TY - JOUR T1 - STAT6 Mediates Footpad Immunopathology in the Absence of IL-12p40 Following Infection of Susceptible BALB/c Mice With . JF - Front Immunol Y1 - 2018 A1 - Florence Kauffmann A1 - Elyn Meert A1 - Kaat de Jonge A1 - Yvon Elkrim A1 - Hanot Mambres, Delphine A1 - Olivier J Denis A1 - Eric Muraille A1 - Magez, Stefan A1 - Carl De Trez AB -

() parasites are intracellular parasites belong to the family and are the causative agent of cutaneous leishmaniasis. This disease affects approximately 1.5 million per year worldwide and there is currently no prophylactic vaccine available. is transmitted by the bite of an infected sandfly and has been considered for decades now as a mouse model of choice to identify the factors implicated in T helper (Th)1 and Th2 polarization due to the natural resistance and susceptibility to infection of C57BL/6 and BALB/c mice, respectively. In this study, we refine the role of IL-12p40 cytokine, which is implicated the development of a protective Th1 response, and STAT6, a transcription factor involved in the signaling detrimental interleukin (IL)-4 and IL-13 associated Th2 cytokines during infection in the BALB/c model. In the absence of STAT6 and IL-12p40 signaling, double knockout (DKO) susceptible BALB/c mice displayed reduced footpad swelling and ulcerative lesion compared to IL-12p40 mice upon infection. Hence, they expressed slower upregulation of keratinocyte markers implicated in the inhibition of wound healing, such as keratin 6a (Krt6a) and Krt16. This coincides with the presence of neutrophils displaying an altered phenotype characterized by a lower expression of surface markers Ly6C, CD11b, and Ly6G. These neutrophils exhibited very lower levels of apoptosis similarly to neutrophils present in resistant STAT6 mice. Interestingly, the reduced footpad swelling in DKO mice is associated with a high footpad parasite level similar to susceptible IL-12p40 mice. In conclusion, this study demonstrate that in the absence of both STAT6 and IL-12p40 signaling, -infected mice display smaller and less ulcerated lesions, which does, however, not correlate with reduced parasite load. In addition, the presence of neutrophils with an altered phenotype is associated with reduced apoptosis and delayed immunopathologies, demonstrating the detrimental role of STAT6 in infected susceptible BALB/c mice.

VL - 9 M3 - 10.3389/fimmu.2018.00503 ER - TY - JOUR T1 - Development of a dot-blot assay for the detection of mould-specific IgE in the Belgian population JF - Mycopathologia Y1 - 2017 A1 - Muriel Vincent A1 - Marta Romano A1 - Corazza,F. A1 - K Huygen A1 - Michel,O. A1 - Olivier J Denis KW - a KW - Belgian KW - Belgian population KW - detection KW - Development KW - IgE KW - POPULATION AB -

Background

Data on mould sensitization in the general population are scarce and mostly on Aspergillus fumigatus, Alternaria alternata and Cladosporium herbarum.

Objectives

To validate a dot-blot assay for the detection of specific IgE and evaluate the prevalence of mould sensitization in a healthy population.

Methods

The dot-blot assay was validated against the CAP test. Sensitization rate to ten common indoor and outdoor mould species in 344 serum samples was calculated. For each serum with more than one reactivity, the “major sensitization” defined as the strongest response against a single mould species was calculated.

Results

Intra- and inter-assay variations were both below 20%, and the positivity threshold of the test was of 0.418 kU/L for A. fumigatus. Correlation with CAP results was strong. The overall prevalence of sensitization was 32.8%, and the commonest sensitizations were against A. alternaria, A. flavus and A. niger (around 15%). The most frequent “major reactivities” were against A. niger and A. alternata (20–30%). In 25.1% of the samples, “major reactivities” were directed against a group of moulds commonly found indoor (Penicillium spp., Aspergillus versicolor, Cladosporium sphaerospermum and Cladosporium cladosporioides).

Conclusions

The dot-blot assay was validated for the detection of mould-specific IgE. In the general population, sensitization to indoor species was common and accounted for 25% of overall mould sensitizations.

VL - 182 CP - 3-4 U1 - 37222 M3 - http://dx.doi.org/10.1007/s11046-016-0091-7 ER - TY - JOUR T1 - Inflammatory Properties and Adjuvant Potential of Synthetic Glycolipids Homologous to Mycolate Esters of the Cell Wall of Mycobacterium tuberculosis. JF - J Innate Immun Y1 - 2017 A1 - H Tima Giresse A1 - Al Dulayymi, Juma''a Raheem A1 - Olivier J Denis A1 - Pauline Lehebel A1 - Baols, Klarah Sherzad A1 - Mohammed Omar Mohsin A1 - L'Homme, Laurent A1 - Sahb, Mohaned Mohammed A1 - Potemberg, Georges A1 - Legrand, Sylvie A1 - Lang, Roland A1 - Beyaert, Rudi A1 - Piette, Jacques A1 - Baird, Mark Stephen A1 - Huygen, Kris A1 - Marta Romano KW - a KW - acid KW - Acids KW - Activation KW - an KW - analysi KW - analysis KW - article KW - AS KW - Belgium KW - Brussels KW - Cell KW - Cell Wall KW - cells KW - Class KW - Cytokines KW - disease KW - Diseases KW - electronic KW - Glucose KW - GMM KW - health KW - Hypothesis KW - Immunisation KW - immunology KW - Infectious KW - Infectious diseases KW - Institute KW - IS KW - journal KW - mechanism KW - mice KW - MODEL KW - Mycobacterium KW - Mycobacterium tuberculosis KW - Mycolic Acids KW - ON KW - Oxygen KW - pathogenic KW - production KW - public KW - public health KW - Public-health KW - Receptor KW - response KW - Responses KW - result KW - results KW - Service KW - Species KW - strain KW - structure KW - study KW - sugar KW - Synthetic KW - System KW - TDM KW - Term KW - Tuberculosis KW - vaccine KW - vaccines KW - WIV-ISP AB -

The cell wall of mycobacteria is characterised by glycolipids composed of different classes of mycolic acids (MAs; alpha-, keto-, and methoxy-) and sugars (trehalose, glucose, and arabinose). Studies using mutant Mtb strains have shown that the structure of MAs influences the inflammatory potential of these glycolipids. As mutant Mtb strains possess a complex mixture of glycolipids, we analysed the inflammatory potential of single classes of mycolate esters of the Mtb cell wall using 38 different synthetic analogues. Our results show that synthetic trehalose dimycolate (TDM) and trehalose, glucose, and arabinose monomycolates (TMM, GMM, and AraMM) activate bone marrow-derived dendritic cells in terms of the production of pro-inflammatory cytokines (IL-6 and TNF-α) and reactive oxygen species, upregulation of costimulatory molecules, and activation of NLRP3 inflammasome by a mechanism dependent on Mincle. These findings demonstrate that Mincle receptor can also recognise pentose esters and seem to contradict the hypothesis that production of GMM is an escape mechanism used by pathogenic mycobacteria to avoid recognition by the innate immune system. Finally, our experiments indicate that TMM and GMM, as well as TDM, can promote Th1 and Th17 responses in mice in an OVA immunisation model, and that further analysis of their potential as novel adjuvants for subunit vaccines is warranted.

VL - 9 CP - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/27855374?dopt=Abstract M3 - 10.1159/000450955 ER - TY - Generic T1 - Development and use of non-invasive biomarkers to monitor respiratory health of young children Y1 - 2016 A1 - S. Nauwelaerts A1 - Koen De Cremer A1 - Olivier J Denis A1 - A. Bernard A1 - Sigrid C.J. De Keersmaecker A1 - Nancy Roosens KW - Biomarkers KW - CHILDREN KW - Development KW - health KW - Institute KW - International KW - Network KW - Respiratory KW - use KW - young JF - Institute Pasteur International Network Scientific Symposium PB - WIV-ISP CY - Paris, France CP - Institut Pasteur U1 - 593 U2 - 29 november-2 december 2016 ER - TY - RPRT T1 - Surveillance des bactéries résistantes aux antibiotiques dans les hôpitaux belges : Rapport Annuel 2014 Y1 - 2015 A1 - Beatrice Jans A1 - Glupczynski, Y A1 - Goossens,H. A1 - Olivier J Denis KW - Antibiotique KW - Antimicrobial resistance KW - Belge KW - CPE KW - ESBL KW - hôpitaux KW - LE MRE KW - MRSA KW - national nosocomial KW - rapport KW - rapport annuel KW - Surveillance KW - Vancomycin Resistance AB -

NA

PB - WIV-ISP CY - Brussels, Belgium ER - TY - Generic T1 - Etude des prévalences et des profils de sensibilisation aux moisissures dans la population belge et dans une population d'asthmatiques par bio-dot Y1 - 2014 A1 - Muriel Vincent A1 - Michel,O. A1 - Corazza,F. A1 - De Prins,S. A1 - K Huygen A1 - Olivier J Denis KW - Belge KW - de KW - ET KW - PAR KW - POPULATION KW - population belge JF - Congrès Francophone d'Allergologie PB - WIV-ISP CY - Paris, France CP - . U1 - 37099 U2 - 17 April 2014 ER - TY - JOUR T1 - Increasing the Vaccine Potential of Live M. bovis BCG by Coadministration with Plasmid DNA Encoding a Tuberculosis Prototype Antigen. JF - Vaccines (Basel) Y1 - 2014 A1 - Nicolas Bruffaerts A1 - Marta Romano A1 - Olivier J Denis A1 - Fabienne Jurion A1 - Huygen, Kris KW - a KW - Activation KW - Adult KW - ALL KW - an KW - antibodies KW - Antibody KW - Antigens KW - approach KW - approaches KW - AS KW - BCG KW - Cell KW - cells KW - CODING KW - Combination KW - concept KW - Control KW - Dna KW - effective KW - Explanation KW - immune response KW - intradermal KW - IS KW - M KW - mice KW - neonate KW - neonates KW - observed KW - ON KW - protocol KW - Pulmonary KW - response KW - Responses KW - result KW - results KW - specific KW - Spleen KW - Still KW - study KW - Tuberculosis KW - Vaccination KW - vaccine KW - Vector AB -

The attenuated live M. bovis Bacille-Calmette-Guérin (BCG) is still the sole vaccine used against tuberculosis, but confers only variable efficacy against adult pulmonary tuberculosis (TB). Though no clear explanation for this limited efficacy has been given, different hypotheses have been advanced, such as the waning of memory T-cell responses, a reduced antigenic repertoire and the inability to induce effective CD8⁺ T-cell responses, which are known to be essential for latent tuberculosis control. In this study, a new BCG-based vaccination protocol was studied, in which BCG was formulated in combination with a plasmid DNA vaccine. As BCG is routinely administered to neonates, we have evaluated a more realistic approach of a simultaneous intradermal coadministration of BCG with pDNA encoding the prototype antigen, PPE44. Strongly increased T- and B-cell responses were observed with this protocol in C57BL/6 mice when compared to the administration of only BCG or in combination with an empty pDNA vector, as measured by Th1-type spleen cell cytokine secretion, specific IgG antibodies, as well as specific IFN-γ producing/cytolytic-CD8⁺ T-cells. Moreover, we observed a bystander activation induced by the coding plasmid, resulting in increased immune responses against other non-plasmid encoded, but BCG-expressed, antigens. In all, these results provide a proof of concept for a new TB vaccine, based on a BCG-plasmid DNA combination.

VL - 2 CP - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26344474?dopt=Abstract M3 - 10.3390/vaccines2010181 ER - TY - JOUR T1 - Mice genetically inactivated in interleukin-17A receptor are defective in long-term control of Mycobacterium tuberculosis infection. JF - Immunology Y1 - 2013 A1 - Freches, Danielle A1 - Korf, Hannelie A1 - Olivier J Denis A1 - Havaux, Xavier A1 - Huygen, Kris A1 - Marta Romano KW - Animals KW - Cytokines KW - Enzyme-Linked Immunosorbent Assay KW - Flow Cytometry KW - mice KW - Mice, Inbred C57BL KW - Mice, Knockout KW - Mycobacterium tuberculosis KW - Receptors, Interleukin-17 KW - Tuberculosis AB -

Interleukin-17A (IL-17A), a pro-inflammatory cytokine acting on neutrophil recruitment, is known to play an important role during Mycobacterium tuberculosis infection, but the role of IL-17A receptor signalling in immune defence against this intracellular pathogen remains poorly documented. Here we have analysed this signalling using C57BL/6 mice genetically inactivated in the IL-17 receptor A subunit (IL-17RA(-/-) ). Although early after infection bacterial growth was controlled to the same extent as in wild-type mice, IL-17RA(-/-) mice were defective in exerting long-term control of M. tuberculosis infection, as demonstrated by a progressively increasing pulmonary bacterial burden and shortened survival time. Compared with infected wild-type mice, IL-17RA(-/-) mice showed impaired recruitment of neutrophils to the lungs at the early but not the late stage of infection. Pulmonary tumour necrosis factor-α, IL-6 and particularly IL-10 levels were decreased in the absence of IL-17RA signalling, whereas IL-1β was increased. CD4(+) -mediated and γδ-mediated IL-17A production was dramatically increased in IL-17RA(-/-) mice (confirming part of their phenotype), whereas production of interferon-γ and expression of the bactericidal enzyme inducible nitric oxide synthase were not affected. Collectively, our data suggest that early but not late neutrophil recruitment is essential for IL-17A-mediated long-term control of M. tuberculosis infection and that a functional interferon-γ response is not sufficient to control M. tuberculosis growth when the IL-17RA pathway is deficient. As treatment of auto-immune diseases with anti-IL-17A antibodies is actually being tested in clinical studies, our data suggest that caution should be taken with respect to possible reactivation of tuberculosis.

VL - 140 CP - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/23721367?dopt=Abstract M3 - 10.1111/imm.12130 ER - TY - JOUR T1 - M.tuberculosis mutants lacking oxygenated mycolates show increased immunogenicity and protective efficacy as compared to M. bovis BCG vaccine in an experimental mouse model. JF - PLoS One Y1 - 2013 A1 - Hedhli, Dorsaf A1 - Olivier J Denis A1 - Barkan, Daniel A1 - Daffé, Mamadou A1 - Glickman, Michael S A1 - Huygen, Kris KW - Animals KW - Bacterial Load KW - BCG Vaccine KW - Disease Models, Animal KW - Enzyme-Linked Immunospot Assay KW - Injections, Subcutaneous KW - Interferon-gamma KW - Interleukin-17 KW - Lymph Nodes KW - mice KW - Mice, Inbred C57BL KW - Mutation KW - Mycobacterium bovis KW - Mycobacterium tuberculosis KW - Mycolic Acids KW - Organ Specificity KW - Oxygen KW - Species Specificity KW - Spleen KW - Th1 Cells KW - Th17 Cells KW - Trachea KW - Treatment Outcome KW - Tuberculosis KW - Vaccination KW - virulence AB -

The existing vaccine against tuberculosis (M. bovis BCG) exerts some protection against the extrapulmonary forms of the disease, particularly in young children, but is not very effective against the pulmonary form of TB, which often results from the reactivation of a latent M. tuberculosis (M.tb)infection. Among the new approaches in TB vaccine development, live attenuated M.tb mutants are a promising new avenue. Here we report on the vaccine potential of two highly attenuated M.tb mutants, MGM1991 and M.tbhma::hyg (HMA), lacking all oxygenated mycolates in their cell wall. In C57BL/6 mice, stronger Th1 (IFN-γ, IL-2 and TNF-α) and IL-17 responses could be induced following subcutaneous vaccination with either of the two mutants, than following vaccination with M. bovis BCG. Significantly more mycobacteria specific IFN-γ producing CD4(+) and particularly CD8(+) T cells could be detected by intracellular cytokine staining in mice vaccinated with the M.tb mutants. Finally, vaccination with either of the two mutants conferred stronger protection against intratracheal M.tb challenge than vaccination with BCG, as indicated by reduced bacterial replication in lungs at 4 to 12 weeks after challenge. Protection against M. tb dissemination, as indicated by reduced bacterial numbers in spleen, was comparable for both mutants to protection conferred by BCG.

VL - 8 CP - 10 U1 - http://www.ncbi.nlm.nih.gov/pubmed/24146869?dopt=Abstract M3 - 10.1371/journal.pone.0076442 ER - TY - JOUR T1 - Characterization of new Alternaria alternata-specific rat monoclonal antibodies JF - Mycopathologia Y1 - 2012 A1 - Olivier J Denis A1 - Van Cauwenberge,A. A1 - Treutens,G. A1 - B. Es Saadi A1 - Symoens,F. A1 - Popovic,N. A1 - K Huygen KW - Alternaria alternata KW - Dwellings KW - ELISA KW - Monoclonal antibodies KW - mould AB -

In this study, three different rat hybridoma cell lines secreting monoclonal antibodies (mAbs) recognizing the spores from Alternaria alternata, a plant pathogenic fungus, contaminant of food products and important cause of both allergic rhinitis and asthma, have been characterized. These three mAbs are all of IgM isotype. Two antibodies, A1 and F10, were cross-reactive antibodies recognizing spores from Alternaria, Cladosporium, Penicillium, Aspergillus and Stachybotrys genera, but not the yeasts Saccharomyces cerevisiae or Candida albicans. Competitive and sandwich assays demonstrated that these two mAbs were directed against the same or very close repetitive(s) epitope(s). A1-based sandwich ELISA efficiently detected this epitope in various mould (but not yeast)-soluble extracts prepared from strains grown in the laboratory. Moreover, this A1-based sandwich ELISA detected its cognate epitope in air and dust samples obtained from dwellings. The third antibody, E5, recognized only the spores of Alternaria and the phylogenetically very close Ulocladium botrytis. This E5 antibody is directed against a repetitive epitope found in Alternaria and Ulocladium laboratory extracts and can be used in a sandwich assay for the quantification of these moulds. Therefore, E5 antibody is a promising tool for the development of Alternaria-Ulocladium-specific immunoassays, while A1 and F10 could be interesting tools for the quantification of the total mould biomass

VL - 173 CP - 2-3 U1 - 38160 M3 - 10.1007/s11046-011-9466-y ER - TY - JOUR T1 - Quantification of the trichothecene Verrucarin-A in environmental samples using an antibody-based spectroscopic biosensor JF - Sensors and Actuators B: Chemical Y1 - 2012 A1 - Gosselin, E. A1 - Olivier J Denis A1 - Van Cauwenberge, A. A1 - Conti, J. A1 - Vanden Eynde, J.J. A1 - Huygen, K. A1 - De Coninck, J. KW - ELISA KW - Fluorescence KW - Fungi KW - mould KW - mycotoxin KW - Spectrometry KW - verrucarin AB -

Verrucarin A2 (VerA) is a toxic trichothecene mycotoxin that can be produced indoors at very low level by moulds contaminating dwellings and may be associated with several human health problems. In this study we describe a spectroscopic label-free biosensor for VerA. This sensor is based on the high sensitivity of Fourier transform infrared-attenuated reflection (FTIR-ATR) spectroscopic detection and the use of a new anti-VerA rat monoclonal antibody (mAb). This antibody was directly grafted at the surface of the infrared element. Competitive ELISA and FTIR-ATR techniques were compared for detection of VerA in buffer and in complex dust samples obtained from dwellings. After optimization, the competitive ELISA showed a sensitivity of 7.43 ng/ml of VerA in PBS and a dynamic range below one order of magnitude. The FTIR technique improved the detection of the VerA by three orders of magnitude (2 pg/ml in buffer and 6 pg/ml when spiked in dust samples). The dynamic range for its detection extended over four orders of magnitude. The percentage of recovery of VerA spiked (1000 ng to 0.1 ng) in a complex dust matrix ranged from 99 to 68%. Our results clearly show that this antibody-based spectroscopic biosensor allow a better detection of VerA as compared to classical immunoassays and can be very efficiently used in the field of indoor mycotoxin detection.

VL - 166-167 M3 - 10.1016/j.snb.2012.03.008 ER - TY - JOUR T1 - Fourier transform infrared immunosensors for model hapten molecules. JF - Biosens Bioelectron Y1 - 2009 A1 - E Gosselin A1 - M Gorez A1 - M Voué A1 - Olivier J Denis A1 - J Conti A1 - N Popovic A1 - A Van Cauwenberg A1 - E Noel A1 - J De Coninck KW - Antibodies, Monoclonal KW - Biosensing Techniques KW - Equipment Design KW - Equipment Failure Analysis KW - Haptens KW - Immunoassay KW - Reproducibility of Results KW - Sensitivity and Specificity KW - Spectroscopy, Fourier Transform Infrared AB -

We report experimental results concerning the detection of 2,4-dinitrophenol, under its free form or coupled to human serum albumin using Fourier transform infrared spectroscopy-based sensors. Competitive immunoreactions were carried out using several anti-dinitrophenol monoclonal antibodies. Comparison with enzyme-linked immunosorbent assays in competition is given for standard operating conditions. FTIR detection limits are comparable to those obtained by ELISA. The limits of detection are about 5-15 ng/mL for the coupled DNP. Using the LO-DNP61 antibody, a detection limit of congruent with 5 ng/mL was also estimated for the free DNP molecules but is much higher for the other antibodies.

VL - 24 CP - 8 M3 - 10.1016/j.bios.2009.01.001 ER - TY - JOUR T1 - Immunogenicity and protective efficacy of tuberculosis DNA vaccines combining mycolyl-transferase Ag85A and phosphate transport receptor PstS-3. JF - Immunology Y1 - 2006 A1 - Marta Romano A1 - Virginie Roupie A1 - Wang, Xiao M A1 - Olivier J Denis A1 - Fabienne Jurion A1 - Adnet, Pierre-Yves A1 - Laali, Rachid A1 - Huygen, Kris KW - Acyltransferases KW - Animals KW - Antigens, Bacterial KW - ATP-Binding Cassette Transporters KW - Bacterial Proteins KW - CD8-Positive T-Lymphocytes KW - Cricetinae KW - Female KW - Immunity, Cellular KW - mice KW - Mice, Inbred BALB C KW - Mice, Inbred C57BL KW - Mycobacterium tuberculosis KW - Plasmids KW - Recombinant Fusion Proteins KW - Th1 Cells KW - Transfection KW - Tuberculosis KW - Tuberculosis Vaccines KW - Vaccines, DNA AB -

DNA vaccines encoding the 32,000 MW mycolyl-transferase Ag85A and the 40,000 MW phosphate-binding protein PstS-3 can elicit protective immune responses against experimental infection with Mycobacterium tuberculosis in C57BL/6 mice. Here we have analysed the vaccine potential of a combination of both antigens using plasmid vectors expressing either a fusion protein of both antigens or the separate proteins driven by two independent promoters (in pBudCE4.1 vector). Comparable levels of Ag85A specific T helper 1 (Th1) type immune responses could be induced by the two combination vaccines and the single vaccine encoding the mycolyl-transferase, whereas induction of PstS-3 specific Th1-mediated responses was impaired in both combination vaccines. In contrast, magnitude of CD8+ mediated responses against the PstS-3 protein was comparable following combination or single DNA vaccination. Antigenic competition was also observed at the antibody level; PstS-3 specific levels being lower in mice vaccinated with the fusion vector and Ag85A specific levels being lower in mice vaccinated with the combination pBudCE4.1 vector (as compared to levels obtained following single plasmid immunization). Protection against M. tuberculosis was only modestly improved in mice vaccinated with the DNA combinations. It is possible that prior activation of Ag85A specific CD4+ T cells directed against this common mycobacterial antigen leads to cross-competition for major histocompatibility complex class II-restricted peptide complexes of the Pst-3 antigen. This may have implications for future combination vaccines using Ag85.

VL - 118 CP - 3 U1 - http://www.ncbi.nlm.nih.gov/pubmed/16827893?dopt=Abstract M3 - 10.1111/j.1365-2567.2006.02373.x ER - TY - JOUR T1 - Mycobacterium tuberculosis with disruption in genes encoding the phosphate binding proteins PstS1 and PstS2 is deficient in phosphate uptake and demonstrates reduced in vivo virulence. JF - Infect Immun Y1 - 2005 A1 - Peirs, Priska A1 - Lefèvre, Philippe A1 - Samira Boarbi A1 - Wang, Xiao-Ming A1 - Olivier J Denis A1 - Braibant, Martine A1 - Pethe, Kevin A1 - Locht, Camille A1 - Huygen, Kris A1 - Content, Jean KW - Animals KW - Bacterial Proteins KW - Culture Media KW - Gene Deletion KW - Humans KW - mice KW - Mice, Inbred BALB C KW - Mice, Inbred C57BL KW - Mycobacterium tuberculosis KW - Phosphate-Binding Proteins KW - Phosphates KW - Tuberculosis KW - virulence AB -

By measuring phosphate uptake by Mycobacterium tuberculosis strains with the pstS1 and pstS2 genes genetically inactivated, we showed that these pstS genes encode high-affinity phosphate binding proteins. In a mouse infection model, both mutants were attenuated in virulence, suggesting that M. tuberculosis encounters limiting phosphate concentrations during its intracellular life span.

VL - 73 CP - 3 U1 - http://www.ncbi.nlm.nih.gov/pubmed/15731097?dopt=Abstract M3 - 10.1128/IAI.73.3.1898-1902.2005 ER - TY - JOUR T1 - Induction of in vivo functional Db-restricted cytolytic T cell activity against a putative phosphate transport receptor of Mycobacterium tuberculosis. JF - J Immunol Y1 - 2004 A1 - Marta Romano A1 - Olivier J Denis A1 - D'Souza, Sushila A1 - Wang, Xiao-Ming A1 - Ottenhoff, Tom H M A1 - Brulet, Jean-Marc A1 - Huygen, Kris KW - Amino Acid Sequence KW - Animals KW - Antibodies, Bacterial KW - ATP-Binding Cassette Transporters KW - Bacterial Proteins KW - Epitopes, T-Lymphocyte KW - H-2 Antigens KW - Histocompatibility Antigen H-2D KW - Interferon-gamma KW - Interleukin-2 KW - mice KW - Mice, Inbred C57BL KW - Molecular Sequence Data KW - Mycobacterium tuberculosis KW - T-Lymphocytes, Cytotoxic KW - Th1 Cells KW - Tuberculosis Vaccines KW - Vaccines, DNA AB -

Using plasmid vaccination with DNA encoding the putative phosphate transport receptor PstS-3 from Mycobacterium tuberculosis and 36 overlapping 20-mer peptides spanning the entire PstS-3 sequence, we determined the immunodominant Th1-type CD4(+) T cell epitopes in C57BL/10 mice, as measured by spleen cell IL-2 and IFN-gamma production. Furthermore, a potent IFN-gamma-inducing, D(b)-restricted CD8(+) epitope was identified using MHC class I mutant B6.C-H-2(bm13) mice and intracellular IFN-gamma and whole blood CD8(+) T cell tetramer staining. Using adoptive transfer of CFSE-labeled, peptide-pulsed syngeneic spleen cells from naive animals into DNA vaccinated or M. tuberculosis-infected recipients, we demonstrated a functional in vivo CTL activity against this D(b)-restricted PstS-3 epitope. IFN-gamma ELISPOT responses to this epitope were also detected in tuberculosis-infected mice. The CD4(+) and CD8(+) T cell epitopes defined for PstS-3 were completely specific and not recognized in mice vaccinated with either PstS-1 or PstS-2 DNA. The H-2 haplotype exerted a strong influence on immune reactivity to the PstS-3 Ag, and mice of the H-2(b, p, and f) haplotype produced significant Ab and Th1-type cytokine levels, whereas mice of H-2(d, k, r, s, and q) haplotype were completely unreactive. Low responsiveness against PstS-3 in MHC class II mutant B6.C-H-2(bm12) mice could be overcome by DNA vaccination. IFN-gamma-producing CD8(+) T cells could also be detected against the D(b)-restricted epitope in H-2(p) haplotype mice. These results highlight the potential of DNA vaccination for the induction and characterization of CD4(+) and particularly CD8(+) T cell responses against mycobacterial Ags.

VL - 172 CP - 11 U1 - http://www.ncbi.nlm.nih.gov/pubmed/15153510?dopt=Abstract ER - TY - JOUR T1 - Purified bovine WC1+ gamma delta T lymphocytes are activated by staphylococcal enterotoxins and toxic shock syndrome toxin-1 superantigens: proliferation response, TCR V gamma profile and cytokines expression. JF - Immunol Lett Y1 - 2001 A1 - Fikri, Y A1 - Olivier J Denis A1 - Pastoret, P A1 - Nyabenda, J KW - Amino Acid Sequence KW - Animals KW - Bacterial Toxins KW - Base Sequence KW - Cattle KW - Cell Division KW - Cell Line KW - Cell Separation KW - Cytokines KW - Enterotoxins KW - Lymphocyte Activation KW - Mitogens KW - Molecular Sequence Data KW - Receptors, Antigen, T-Cell, gamma-delta KW - RNA, Messenger KW - Staphylococcus aureus KW - Superantigens KW - T-Lymphocyte Subsets AB -

In this study, the ability of purified bovine gammadelta T cells in vitro to be activated by superantigens (SAg) was investigated. Freshly isolated WC1(+) gammadelta T cells, in the presence of autologous glutaraldehyde-fixed or gamma-irradiated antigen presenting cells (APC) and IL-2, were incubated with staphylococcal enterotoxins A and B (SEA and SEB), and toxic shock syndrome toxin-1 (TSST-1). Both a proliferative response and the expression of particular T cell receptor genes of the gamma variable (TCR Vgamma) repertoire family were induced. Genes encoding TCR Vgamma1 and TCR Vgamma2 family, but not TCR Vgamma5 were detected. The cells also expressed cytokine transcripts, namely, those of IL-12, IFN-gamma and TNF-alpha, but not IL-2, IL-4, IL-6, IL-7 and IL-10. The activation and proliferation of freshly isolated gammadelta T cells by non-processed antigens required two signals, one originating from the APC and a second dependent on exogenous IL-2. Our results show that purified bovine WC1(+) gammadelta T cells could be driven to proliferate and to express a particular TCRVgamma profile in response to superantigen activation. The possible implication of cytokines expressed by bovine gammadelta T cells in immunopathogenesis is discussed.

VL - 77 CP - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/11377702?dopt=Abstract ER - TY - JOUR T1 - What are the immunologically active components of bacille Calmette-Guérin in therapy of superficial bladder cancer? JF - Int J Cancer Y1 - 2000 A1 - A R Zlotta A1 - Van Vooren, J P A1 - Olivier J Denis A1 - Drowart, A A1 - M Daffé A1 - Lefèvre, P A1 - L Schandene A1 - De Cock, M A1 - De Bruyn, J A1 - Vandenbussche, P A1 - Fabienne Jurion A1 - Palfliet, K A1 - J Simon A1 - C C Schulman A1 - Content, J A1 - Huygen, K KW - Antigens, Bacterial KW - Antigens, CD KW - Bacterial Outer Membrane Proteins KW - BCG Vaccine KW - CD56 Antigen KW - Humans KW - Interferon-gamma KW - Interleukin-12 KW - Interleukin-2 KW - Interleukin-6 KW - Leukocytes, Mononuclear KW - Neoplasm Proteins KW - Th1 Cells KW - Tumor Cells, Cultured KW - Urinary Bladder Neoplasms AB -

The subcomponents of bacille Calmette-Guérin (BCG) involved in the mechanism of action of intravesical BCG immunotherapy used for prophylaxis of superficial bladder cancer recurrences have been poorly investigated. We purified various BCG subcomponents and analyzed in vitro their ability to enhance a Th1 polarized immune response as well as to increase lymphocyte-mediated cytotoxicity against bladder tumors. Human peripheral blood mononuclear cells (PBMCs) from healthy purified protein derivative-positive subjects were incubated for 7 days with whole BCG and various fractions (BCG cell wall, plasma membrane, cytosol, purified polysaccharides as glucan or arabinomannan, purified native proteins from BCG culture filtrate, recombinant 22 kDa protein, phosphate transporter PstS-2 and -3 proteins). IFN-gamma, IL-12, IL-2, and IL-6 production by stimulated PBMCs was compared to unstimulated controls and the phenotype of expanded cells analyzed by flow cytometry (FACS analysis). A (51)Cr-release assay monitored the cytotoxicity of amplified effector cells against T24 bladder tumor cells. Live BCG and most of its subcomponents (with the exception of cytosol, PstS-2 and -3) significantly enhanced IFN-gamma and IL-12 secretion, expanded CD3(-)CD56(+) cells and the non-MHC-restricted cytotoxicity against bladder tumor cells compared to unstimulated controls (all P < 0.001, t-test). IL-2 receptor blockage resulted in a clear reduction in the cytotoxic activity of stimulated PBMCs. Numerous BCG subcomponents thus provide positive stimuli for Th1 cell differentiation and enhance in vitro, non-MHC-restricted cytotoxicity against bladder tumor cells. Our findings provide the basis for the therapeutic use of several of these subfractions in experimental animal models bearing bladder tumors.

VL - 87 CP - 6 ER - TY - JOUR T1 - What is the optimal regimen for BCG intravesical therapy? Are six weekly instillations necessary? JF - Eur Urol Y1 - 2000 A1 - A R Zlotta A1 - Van Vooren, J P A1 - Huygen, K A1 - Drowart, A A1 - M Decock A1 - M Pirson A1 - Fabienne Jurion A1 - Palfliet, K A1 - Olivier J Denis A1 - J Simon A1 - C C Schulman KW - Administration, Intravesical KW - Aged KW - Antigens, Bacterial KW - BCG Vaccine KW - Carcinoma, Transitional Cell KW - Cystoscopy KW - Dose-Response Relationship, Drug KW - Drug Administration Schedule KW - Female KW - Follow-Up Studies KW - Humans KW - Immunotherapy KW - Lymphocyte Count KW - Male KW - middle aged KW - Mycobacterium bovis KW - T-Lymphocytes KW - Urinary Bladder Neoplasms AB -

OBJECTIVE: For more than 20 years, BCG intravesical therapy schedule has included 6 weekly instillations. Very few studies have, however, analyzed the rationale of this regimen. We previously demonstrated that intravesical BCG induced an increased peripheral immune response against mycobacterial antigens as compared to pretreatment values. In the present work, we have studied the weekly evolution of this immune response induced by intravesical BCG instillations.

MATERIALS AND METHODS: The evolution of the lymphoproliferative response of peripheral blood mononuclear cells against BCG culture filtrate (CF), tuberculin (PPD) and BCG extract (EXT) was tested before, every week during the BCG instillations and at 3 and 6 months follow-up in 9 patients with superficial bladder cancer treated with 6 weekly BCG instillations. Lymphoproliferation was measured by means of a tritiated thymidine incorporation test.

RESULTS: A significant increase in the lymphoproliferative response against PPD, CF and EXT was observed in 9, 8 and 7 of the 9 patients, respectively, as compared to pre-BCG values. The maximal lymphoproliferation was achieved after 4 instillations in 4/5 patients initially reactive against mycobacterial antigens whereas 2 of 4 initially nonreactive patients required 6 instillations. At 6 months' follow-up, lymphoproliferation against BCG and the other mycobacterial antigens returned to pre-BCG values in all patients. In 3 patients who received additional instillations because of tumor recurrence within 1 year of follow-up, the maximum immune response was observed already after 2 instillations.

CONCLUSION: In most patients, the maximal peripheral immune response is already observed after 4 weekly instillations. However, patients not previously immunized against mycobacterial antigens may require 6 weekly instillations to achieve a maximum stimulation level. Our data support the need to further evaluate the role of this status before starting BCG instillations. It could be of interest to study whether 6 BCG instillations are really necessary in patients previously immune against mycobacterial antigens.

VL - 37 CP - 4 M3 - 10.1159/000020170 ER - TY - JOUR T1 - Characterization of the culture filtrate-specific cytotoxic T lymphocyte response induced by bacillus Calmette-Guérin vaccination in H-2b mice JF - International Immunology Y1 - 1999 A1 - Olivier J Denis A1 - Huygen, Kris KW - BCG Mycobacterium bovis bacille Calmette-Guérin KW - BFA Brefeldin A KW - CF culture filtrate KW - CTL cytotoxic T lymphocytes KW - culture filtrate antigens KW - cytotoxic T lymphocytes KW - Mycobacterium bovis bacillus Calmette-Guérin KW - TAP transporter-associated protein KW - Tuberculosis AB -

Although CD8+ T cells are supposed to play an important role in protective immunity to mycobacteria, cytotoxic T lymphocyte (CTL) responses in this infection remain poorly characterized. We previously demonstrated that bacillus Calmette-Guérin (BCG) immunization of H-2b mice induced CTL able to recognize and kill macrophages incubated with proteins from mycobacterial culture supernatant [culture filtrate (CF) antigens]. In the present study, we have further characterized the lytic activity of these CTL and the processing pathway used for the presentation of CF proteins. We show that they use the degranulation pathway (secretion of perforins and granzymes) as the main lytic mechanism of cytotoxicity and also secrete IFN-γ upon incubation with CF-pulsed macrophages. The in vitro presentation of CF proteins to CTL required a processing step inhibited in the cold but insensitive to Brefeldin A. Transporter-associated protein (TAP)-2-deficient RMA-S cells were efficiently recognized and killed by CF-specific CTL, demonstrating the lack of TAP requirement for this presentation. However, recognition of target cells by CTL was abolished when carried out in the presence of chloroquine. These results indicate that a non-classical MHC class I-processing pathway allows the recognition of a CF protein by CTL in BCG-vaccinated H-2b mice.

VL - 11 CP - 2 M3 - 10.1093/intimm/11.2.209 ER - TY - JOUR T1 - Immunogenicity and protective efficacy of tuberculosis DNA vaccines encoding putative phosphate transport receptors. JF - J Immunol Y1 - 1999 A1 - Tanghe, A A1 - Lefèvre, P A1 - Olivier J Denis A1 - D'Souza, S A1 - Braibant, M A1 - Lozes, E A1 - Singh, M A1 - Montgomery, D A1 - Content, J A1 - Huygen, K KW - Animals KW - Antibodies, Bacterial KW - Antigens, Bacterial KW - ATP-Binding Cassette Transporters KW - Bacterial Proteins KW - Bacterial Vaccines KW - Cross Reactions KW - Escherichia coli Proteins KW - Female KW - Interleukin-2 KW - Lung KW - mice KW - Mice, Inbred C57BL KW - Mycobacterium KW - Mycobacterium tuberculosis KW - Periplasmic Binding Proteins KW - Phosphate-Binding Proteins KW - Phosphates KW - Plasmids KW - Spleen KW - Tuberculosis KW - Vaccines, DNA AB -

Using culture filtrate Ag-specific mAbs generated from mycobacteria-infected H-2b haplotype mice, we have previously identified three genes in the Mycobacterium tuberculosis genome, encoding proteins homologous to the periplasmic ATP-binding cassette phosphate-binding receptor PstS of the phosphate-specific transport system of E. coli. To define the potential vaccinal properties of these phosphate-binding proteins, female C57BL/6 mice were injected i.m. with plasmid DNA encoding PstS-1, PstS-2, or PstS-3 proteins from M. tuberculosis and immunogenicity and protective efficacy against i.v. challenge with M. tuberculosis H37Rv was analyzed. Significant levels of highly Ag-specific Abs and Th1-type cytokines IL-2 and IFN-gamma could be detected following vaccination with each of the three genes. However, only mice vaccinated with PstS-3 DNA demonstrated significant and sustained reduction in bacterial CFU numbers in spleen and lungs for 3 mo after M. tuberculosis challenge, as compared with CFU counts in mice vaccinated with control DNA. Vaccination with PstS-2 DNA induced a modest reduction in CFU counts in spleen only, whereas vaccination with PstS-1 DNA was completely ineffective in reducing bacterial multiplication. In conclusion, our results indicate that DNA vaccination is a powerful and easy method for comparative screening of potentially protective Ags from M. tuberculosis and that the PstS-3 protein is a promising new subunit vaccine candidate.

VL - 162 CP - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/9916741?dopt=Abstract ER - TY - JOUR T1 - DNA vaccines against tuberculosis. JF - Novartis Found Symp Y1 - 1998 A1 - Ulmer, J B A1 - Montgomery, D L A1 - Tang, A A1 - Zhu, L A1 - Deck, R R A1 - C DeWitt A1 - Olivier J Denis A1 - I Orme A1 - Content, J A1 - Huygen, K KW - Acyltransferases KW - Animals KW - Antibodies, Bacterial KW - Antigens, Bacterial KW - BCG Vaccine KW - Female KW - Guinea Pigs KW - Immunity, Cellular KW - mice KW - Mice, Inbred BALB C KW - Tuberculosis, Pulmonary KW - Vaccination KW - Vaccines, DNA AB -

DNA plasmids encoding Mycobacterium tuberculosis antigen 85 (Ag85) were tested as vaccines in animal models. Ag85 DNA induced relevant immune responses (i.e. T helper (Th) cells, Th1 cytokines and cytotoxic T lymphocytes) and was protective in mouse and guinea pig models of mycobacterial disease. Therefore, DNA vaccination holds promise as an effective means of preventing tuberculosis in humans. Furthermore, this technique is amenable to identifying the protective antigens of M. tuberculosis.

VL - 217 ER - TY - JOUR T1 - Vaccination with plasmid DNA encoding mycobacterial antigen 85A stimulates a CD4+ and CD8+ T-cell epitopic repertoire broader than that stimulated by Mycobacterium tuberculosis H37Rv infection. JF - Infect Immun Y1 - 1998 A1 - Olivier J Denis A1 - Tanghe, A A1 - Palfliet, K A1 - Fabienne Jurion A1 - Thierry van den Berg A1 - Vanonckelen, A A1 - Ooms, J A1 - Saman, E A1 - Ulmer, J B A1 - Content, J A1 - Huygen, K KW - Amino Acid Sequence KW - Animals KW - Antigens, Bacterial KW - Bacterial Vaccines KW - CD4-Positive T-Lymphocytes KW - CD8-Positive T-Lymphocytes KW - Cross Reactions KW - Epitopes, T-Lymphocyte KW - Interferon-gamma KW - Interleukin-2 KW - mice KW - Mice, Inbred BALB C KW - Molecular Sequence Data KW - Mycobacterium tuberculosis KW - T-Lymphocytes, Cytotoxic KW - Vaccination KW - Vaccines, DNA AB -

Vaccination of mice with plasmid DNA carrying the gene for the major secreted mycobacterial antigen 85A (Ag85A) from Mycobacterium tuberculosis is a powerful technique for generating robust specific Thl helper T-cell responses, CD8+-mediated cytotoxicity, and protection against M. tuberculosis challenge (K. Huygen et al., Nat. Med. 2:893-898, 1996). We have now analyzed in more detail the antigen-specific immune CD4+- and CD8+-T-cell responses induced in BALB/c mice vaccinated with Ag85A DNA and have compared these responses to those generated by intravenous infection with M. tuberculosis. T-cell-epitope mapping, as measured by interleukin-2 and gamma interferon secretion from splenic T cells restimulated in vitro with synthetic 20-mer peptides spanning the complete mature sequence of Ag85A, demonstrated that DNA vaccination stimulated a stronger and broader T-cell response than did M. tuberculosis infection. Moreover, elevated cytotoxic T lymphocyte (CTL) activity against Ag85A-transfected and peptide-pulsed P815 target cells could be generated exclusively by vaccination with plasmid DNA, not following M. tuberculosis infection. By using DNA vaccination, three Ag85A CTL epitopes with predicted major histocompatibility complex class I binding motifs were defined. One of them was previously reported as a dominant, promiscuously recognized T-cell epitope in healthy humans with primary infections. These data strengthen the potential of DNA vaccination with respect to inducing antituberculous immunity in humans.

VL - 66 CP - 4 U1 - http://www.ncbi.nlm.nih.gov/pubmed/9529077?dopt=Abstract ER - TY - JOUR T1 - Humoral response against heat shock proteins and other mycobacterial antigens after intravesical treatment with bacille Calmette-Guérin (BCG) in patients with superficial bladder cancer. JF - Clin Exp Immunol Y1 - 1997 A1 - A R Zlotta A1 - Drowart, A A1 - Huygen, K A1 - De Bruyn, J A1 - H Shekarsarai A1 - M Decock A1 - M Pirson A1 - Fabienne Jurion A1 - Palfliet, K A1 - Olivier J Denis A1 - F Mascart A1 - J Simon A1 - C C Schulman A1 - Van Vooren, J P KW - Administration, Intravesical KW - Aged KW - Antibodies, Bacterial KW - Antibody Specificity KW - Antigens, Bacterial KW - BCG Vaccine KW - Cell Division KW - Chaperonin 60 KW - Escherichia coli KW - Escherichia coli Proteins KW - Female KW - Heat-Shock Proteins KW - HSP70 Heat-Shock Proteins KW - Humans KW - Immunodominant Epitopes KW - Immunoglobulin G KW - Immunoglobulin M KW - Immunotherapy KW - Lymphocytes KW - Male KW - middle aged KW - Mycobacterium KW - Mycobacterium bovis KW - Recombinant Proteins KW - Tetanus Toxoid KW - Tuberculin KW - Urinary Bladder Neoplasms AB -

Few studies have analysed the antibody response during intravesical BCG immunotherapy for superficial bladder cancer. We have examined the evolution in serum antibody response against several heat shock proteins (hsp), including the recombinant mycobacterial hsp65 and the native protein P64 from BCG, GroEL from Escherichia coli (hsp60 family), recombinant mycobacterial hsp70 and the E. coli DnaK (hsp70 family), against purified protein derivative of tuberculin (PPD) and the AG85 complex of Mycobacterium bovis BCG, as well as against tetanus toxoid in 42 patients with a superficial bladder tumour, 28 treated with six intravesical BCG instillations and 14 patients used as controls. We also analysed the lymphoproliferative response of peripheral blood mononuclear cells against PPD in this population. Data of antibody responses at 6 weeks post BCG were available in all 28 patients, and at 4 month follow up in 17 patients. All patients who demonstrated a significant increase in IgG antibodies against PPD at 4 months follow up had a significant increase already at 6 weeks of follow up. In contrast, IgG antibodies against hsp increased significantly from 6 weeks to 4 months post-treatment. A significant increase in IgG antibodies against PPD, hsp65, P64, GroEL, and hsp70 at 4 months follow up was observed in 10/17, 8/17, 10/17, 4/17 and 8/17 patients. Native P64 protein elicited a higher antibody response than recombinant mycobacterial hsp65. No increase in antibody response was observed against Dnak from E. coli, against AG85 or tetanus toxoid after BCG therapy. An increase in IgG antibodies against P64 at 4 months follow up compared with pretreatment values was found to be a significant predictor of tumour recurrence (P<0.01). Further studies with a larger number of patients are needed to confirm the value of the antibody response against P64 as a clinical independent prognostic factor.

VL - 109 CP - 1 ER - TY - JOUR T1 - Induction of cytotoxic T-cell responses against culture filtrate antigens in Mycobacterium bovis bacillus Calmette-Guérin-infected mice. JF - Infect Immun Y1 - 1997 A1 - Olivier J Denis A1 - Lozes, E A1 - Huygen, K KW - Animals KW - Antigen Presentation KW - Antigens, Bacterial KW - BCG Vaccine KW - CD4-Positive T-Lymphocytes KW - CD8-Positive T-Lymphocytes KW - Cell-Free System KW - Cross Reactions KW - Culture Media KW - Cytotoxicity, Immunologic KW - Epitopes KW - Female KW - H-2 Antigens KW - Interleukin-2 KW - Lymphocyte Activation KW - mice KW - Mice, Inbred BALB C KW - Mice, Inbred C3H KW - Mice, Inbred C57BL KW - Mice, Inbred DBA KW - Mycobacterium bovis KW - T-Lymphocytes, Cytotoxic AB -

CD8+ T cells are essential for protection against mycobacteria, as is clearly demonstrated by the fatal outcome of experimental infection of beta-2 microglobulin knockout mice. However, the mechanisms and antigens (Ags) leading to CD8+ T-cell activation and regulation have been poorly characterized. Here we show that, upon immunization of major histocompatibility complex (MHC)-congenic mice with Mycobacterium bovis bacillus Calmette-Guérin (BCG), a cytotoxic response against BCG culture filtrate (CF) Ags (CFAgs) is induced in H-2b and H-2bxd haplotypes but not in H-2d haplotype. This response is mediated by CD8+ T cells and absolutely requires the activation of CD4+ T cells and their secretion of interleukin 2. The lack of cytotoxic response in H-2d mice cannot be explained by impaired cytokine production or by a defect in Ag presentation by H-2d macrophages. Using the MHC class I mutant B6.C-H-2bm13 mouse strain, we demonstrate that cytotoxic T lymphocytes (CTLs) recognize CFAgs exclusively in association with D(b) molecules. These Ags are cross-reactive in mycobacteria, since BCG-induced CTLs also recognize macrophages pulsed with CF from Mycobacterium tuberculosis H37Rv and H37Ra and from two virulent strains of M. bovis. Moreover, immunization with Mycobacterium kansasii induces CTLs able to lyse macrophages pulsed with BCG CF. Finally, we have found that these Ags can be characterized as hydrophilic proteins, since they do not bind to phenyl-Sepharose CL-4B. Our results indicate that MHC-linked genes exert a profound influence on the generation of CD8+ CTLs following BCG vaccination.

VL - 65 CP - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/9009330?dopt=Abstract ER - TY - JOUR T1 - Induction of immunity by DNA vaccination: application to influenza and tuberculosis. JF - Behring Inst Mitt Y1 - 1997 A1 - Ulmer, J B A1 - Deck, R R A1 - C M De Witt A1 - J J Donnelly A1 - A Friedman A1 - Montgomery, D L A1 - Yawman, A M A1 - Orme, I M A1 - Olivier J Denis A1 - Content, J A1 - Huygen, K A1 - Liu, M A KW - Animals KW - Antibodies, Bacterial KW - Antibodies, Viral KW - Antibody Formation KW - Antigens, Bacterial KW - Antigens, Viral KW - BCG Vaccine KW - Cytokines KW - Immunity, Cellular KW - Influenza A virus KW - Influenza Vaccines KW - Lymphocyte Activation KW - mice KW - Mice, Inbred BALB C KW - Mice, Inbred C57BL KW - Mycobacterium tuberculosis KW - Nucleoproteins KW - RNA-Binding Proteins KW - Spleen KW - T-Lymphocytes, Cytotoxic KW - Vaccines, DNA KW - Viral Core Proteins AB -

DNA vaccination is an effective means of inducing both humoral and cell-mediated immunity in animal models of infectious disease. Presented here are data generated in two distinct disease models; one viral (influenza) and one bacterial (tuberculosis). Specifically, plasmid DNA encoding an influenza virus antigen (nucleoprotein; NP) and a Mycobacterium tuberculosis antigen (antigen 85; Ag85) were prepared and tested as DNA vaccines in mice. In both cases, high titer antibody responses and robust cell-mediated immune responses were induced against the respective antigens. With respect to the latter, lymphocyte proliferation, Th1-type cytokine secretion, and cytotoxic T lymphocyte responses were observed upon restimulation with antigen in vitro. Furthermore, protective efficacy in animal challenge models was demonstrated in both systems. The data support the hypothesis that DNA vaccination will prove to be a broadly applicable technique for inducing immunity against various infectious diseases.

CP - 98 ER - TY - JOUR T1 - The perinatal presence of antigen (p-azophenylarsonate) or anti-mu antibodies lead to the loss of the recurrent idiotype (CRIA) in A/J mice. JF - Int Immunol Y1 - 1995 A1 - Ryelandt, M A1 - D De Wit A1 - A Baz A1 - Vansanten, G A1 - Olivier J Denis A1 - F Huetz A1 - Nisol, F A1 - Macedo-Soares, F A1 - S Barcy A1 - Brait, M KW - Animals KW - Animals, Newborn KW - Antibodies, Anti-Idiotypic KW - Antibody Affinity KW - gamma-Globulins KW - Immune Tolerance KW - Immunoglobulin Idiotypes KW - Immunoglobulin M KW - Lymphocyte Count KW - mice KW - Mice, Inbred A KW - p-Azobenzenearsonate KW - Stem Cells AB -

The immune response of A/J mice against p-azophenylarsonate (Ars)-keyhole limpet hemocyanin (KLH) is characterized by the dominance, late in primary and during the secondary, of a recurrent idiotype called CRIA, encoded by a canonical combination of Ig gene segments. In this study, A/J mice were given Ars coupled to deaggregated human gamma globulins (dHGG) within 24 h after delivery. The offsprings from these mice were then exposed as adults to Ars-KLH. These animals developed an unusual immune response. The level of anti-Ars antibodies was nearly normal but a dramatic shift in repertoire was observed: the cross-reactive idiotype which is the hallmark of the anti-Ars response in A/J mice was completely absent. The idiotype could be recovered by injection of anti-idiotypic antibodies alone, with no need of lipopolysaccharide coupling. Therefore the presence of antigen at birth can lead to a strong perturbation of idiotype selection. Similar results were obtained with neonatal treatment using anti-IgM antibodies. After recovery of suppression, A/J mice can mount an anti-arsonate response of normal level but devoid of the dominant idiotype.

VL - 7 CP - 4 ER - TY - JOUR T1 - Achievement of serum immunoglobulin depletion in rodents by injections of anti-immunoglobulin antibodies. JF - Transplant Proc Y1 - 1994 A1 - Latinne, D A1 - Olivier J Denis A1 - M Soares A1 - Bazin, H KW - Animals KW - Antibodies, Anti-Idiotypic KW - Immunoglobulins KW - mice KW - rats KW - Rodentia VL - 26 CP - 2 ER -