Charles-Antoine COUPET - Academia.edu (original) (raw)

Papers by Charles-Antoine COUPET

Human Vaccines & Immunotherapeutics

<p>Groups of mice co-treated with antibiotics (ATB) and MVATG18598 or MVATGN33.1 given thri... more <p>Groups of mice co-treated with antibiotics (ATB) and MVATG18598 or MVATGN33.1 given thrice subcutaneously during antibiotic therapy were analyzed.</p

<p>Three antigen fusions were inserted in the deletion III of MVA vector. The first fusion ... more <p>Three antigen fusions were inserted in the deletion III of MVA vector. The first fusion is constituted by the fusion of Rv2029, Rv2626, Rv1733 and Rv0111 proteins and is placed under the control of p7.5K promoter. The second fusion contains a fusion of RpfB-RpfD, Ag85B, TB10.4 and ESAT-6 proteins and its expression is driven by the pH5R promoter. The third fusion is constituted by the fusion of Rv0569, Rv1813, Rv3407, Rv3478 and Rv1807 proteins and is placed under the control of pB2R promoter. SF, signal peptide of the F protein of measles virus. SR, signal peptide of the glycoprotein precursor of rabies virus ERA strain. TMR, membrane-anchoring peptide derived from the rabies glycoprotein of PG strain.</p

<p>Cells from MVATG18377-immunized BALB/c mice were stimulated with antigen peptide pools o... more <p>Cells from MVATG18377-immunized BALB/c mice were stimulated with antigen peptide pools or an irrelevant E7 peptide (Irr) and IFNγ, IL2 and TNFα intracellular cytokine staining was measured by flow cytometry. Results are presented as (<b>A</b>) the percentage of IFNγ⁺, IFNγ⁺TNFα⁺ or IFNγ⁺TNFα⁺IL2⁺ cell subsets among total CD4 T cells or (<b>B</b>) percentage of IFNγ⁺, TNFα⁺ or IFNγ⁺TNFα⁺ cell subsets among total CD8 T cells. Plain bars represent response from individual mice and hatched bars represent median response for each cell subset. Cut-off value (dotted line, 0.02%) is represented for both CD4⁺ and CD8⁺ T cell responses. Only antigens with median values above the cut-off value are represented. For these antigens, only cell subgroups with a percentage above the cut-off value are represented. No response was detected in MVATGN33.1-immunized mice (data not shown). For each cell population, background signal obtained in unstimulated cell condition was subtracted. Pie charts represent a more global analysis for each responder antigen. All analyzed single (IFNγ⁺, TNFα⁺ and IL2⁺), double (IFNγ⁺TNFα⁺, IFNγ⁺IL2⁺ and TNFα⁺IL2⁺) or triple (IFNγ⁺TNFα⁺IL2⁺) cytokine producer cells are included under the corresponding color codes. Results are representative of two independent experiments.</p

<p><b>(A)</b> Scheme of immunotherapy experiments. BALB/c mice were infected wi... more <p><b>(A)</b> Scheme of immunotherapy experiments. BALB/c mice were infected with a low dose aerosol of <i>M</i>. <i>tuberculosis</i> H37Rv. Four weeks later, mice were treated with an antibiotic regimen (RHZ) for 11 weeks. A subset of antibiotic-treated mice was vaccinated, through subcutaneous (SC) or intranasal (IN) routes, three times with a 3-week interval, with either MVATGN33.1 or MVATG18598 during (Weeks 7, 10 and 13) or after (Weeks 17, 20 and 23) chemotherapy. Twelve weeks after the end of the antibiotic regimen (Week 27), the number of viable bacteria in the lungs of animals was determined. <b>(B)</b> <i>M</i>. <i>tuberculosis</i> CFU counts after the completion of the antibiotic regimen in mice vaccinated or not (black) with either MVATGN33.1 (grey) or MVATG18598 (orange). Each symbol represents CFU value of individual mice. Dotted line indicates the lower limit of detection. Results are expressed as mean (black line) log₁₀ CFU ± S.E.M. Statistical analysis was performed using a Mann-Whitney test. *, <i>p</i><0.05. <b>(C)</b> Titers of antibodies specific to MVATG18598-expressed antigens at the relapse evaluation. Each symbol represents antibody titer (log₁₀) of individual mice vaccinated or not (black) with either MVATGN33.1 (grey) or MVATG18598 (orange). Black line represents median value for each group. Statistical analysis was performed using a Mann-Whitney test. §, <i>p</i><0.05 and §§, <i>p</i><0.01 for comparison with RHZ group. *, <i>p</i><0.05 and **, <i>p</i><0.01 for comparison between MVATGN33.1- and MVATG18598-vaccinated groups.</p

<p>Groups of mice co-treated with antibiotics (ATB) and MVATG18598 or MVATGN33.1 given thri... more <p>Groups of mice co-treated with antibiotics (ATB) and MVATG18598 or MVATGN33.1 given thrice subcutaneously during antibiotic therapy were analyzed.</p

La tuberculose (TB), maladie pulmonaire causée par le Mycobacterium tuberculosis (Mtb), reste la ... more La tuberculose (TB), maladie pulmonaire causée par le Mycobacterium tuberculosis (Mtb), reste la première cause de mortalité par un agent infectieux. La TB est responsable de près de 1,7 million de morts et de 10,4 millions de nouveaux cas par an dans le monde. L’émergence et la propagation de souches bactériennes multi-résistantes aux antibiotiques (MDR) représentent une menace majeure grandissante et reflètent l’efficacité partielle des thérapies actuelles. Le traitement des patients atteints de TB-MDR est constitué actuellement de combinaisons d’antibiotiques, souvent toxiques, administrés pendant une longue durée, avec une efficacité limitée. Il existe donc un besoin urgent de développer de nouveaux traitements antituberculeux. L’immunothérapie, dont l’objectif est d’améliorer la réponse immunitaire de l’hôte contre le Mtb, représente une approche complémentaire intéressante dans le but de diminuer la durée et d’augmenter l’efficacité des traitements actuels de la TB-MDR. Le pre...

<p>Mice were immunized twice with either MVATGN33.1 or MVATG18377. Labelled target cell pop... more <p>Mice were immunized twice with either MVATGN33.1 or MVATG18377. Labelled target cell populations pulsed with individual antigen peptide pool were adoptively transferred into those 7 days after last immunization. Results are expressed as mean ± S.D. of the percentage of specific killing based on the relative ratio of target cells present in MVATG18377-vaccinated mice compared to those in the MVATGN33.1-vaccinated mice. For each time point and antigen peptide pool (P), significant lysis is indicated by * (p<0.05) using a permutation resampling test.</p

<p>Cells from MVATG18377-immunized C57BL/6 mice were stimulated with antigen peptide pools ... more <p>Cells from MVATG18377-immunized C57BL/6 mice were stimulated with antigen peptide pools or an irrelevant E7 peptide (Irr). Results are presented as (<b>A</b>) the percentage of IFNγ⁺, IFNγ⁺TNFα⁺ or IFNγ⁺TNFα⁺IL2⁺ cell subsets among total CD4 T cells or (<b>B</b>) percentage of IFNγ⁺, TNFα⁺ or IFNγ⁺TNFα⁺ cell subsets among total CD8 T cells. Plain bars represent response from individual mice and hatched bars represent median response for each cell subset. Cut-off value (dotted line, 0.02%) is represented for both CD4⁺ and CD8⁺ T cell responses. Only antigens with median value above the cut-off value are represented. For these antigens, only cell subgroups with a percentage above the cut-off value are represented. No response was detected in MVATGN33.1-immunized mice (data not shown). For each cell population, background signal obtained in unstimulated cell condition was subtracted. Pie charts represent a more global analysis for each responder antigen. All analyzed single (IFNγ⁺, TNFα⁺ and IL2⁺), double (IFNγ⁺TNFα⁺, IFNγ⁺IL2⁺ and TNFα⁺IL2⁺) or triple (IFNγ⁺TNFα⁺IL2⁺) cytokine producer cells are included under the corresponding color codes. Results are representative of two independent experiments.</p

<p>Percentage of relapsing mice and bacterial burden in mice vaccinated SC with increasing ... more <p>Percentage of relapsing mice and bacterial burden in mice vaccinated SC with increasing vaccination number of MVATG18598 during antibiotic regimen.</p

<p><b>(A)</b> MVATG18598 contains the fusion Rv2626/T2a/Ag85B under the control... more <p><b>(A)</b> MVATG18598 contains the fusion Rv2626/T2a/Ag85B under the control of pB2R promoter, the fusion CFP10/ESAT6 under the control of pH5R promoter, the fusion TB10.4/Rv0287 under the control of pSE/L promoter, the fusion RpfB-RpfD under the control of p7.5K promoter and the fusion Rv3407/E2a/Rv1813 under the control of pA35R promoter. <b>(B)</b> CEF cells were infected or not (Mock) with MVATG18598 or MVATGN33.1 and cell extracts analyzed by Western blot. Fusion Rv2626/T2a/Ag85B was detected using a mouse monoclonal anti-Rv2626 antibody (expected molecular weight of cleaved product: 17.4 kDa) and a rabbit polyclonal anti-Ag85B antibody (expected molecular weight of cleaved product: 31.3 kDa). CFP10/ESAT6 fusion (expected molecular weight: 21.8 kDa) was detected using a mouse monoclonal anti-ESAT6 antibody. TB10.4/Rv0287 fusion (expected molecular weight: 21.2 kDa) was detected using an anti-TB10.4 rabbit polyclonal antibody. Fusion RpfB-RpfD (expected molecular weight: 37.0 kDa) was detected using a rabbit polyclonal anti-RpfB antibody. Fusion Rv3407/E2a/Rv1813 was detected using a rabbit polyclonal anti-Rv3407 antibody (expected molecular weight of cleaved product: 13.2 kDa) and a rabbit polyclonal anti-Rv1813 antibody (expected molecular weight of cleaved product: 12.1 kDa). ND; not done.</p

<p>(<b>A</b>) BALB/c, (<b>B</b>) C57BL/6 and (<b>C</b>)... more <p>(<b>A</b>) BALB/c, (<b>B</b>) C57BL/6 and (<b>C</b>) C3H/HeN mice were immunized once with either MVATGN33.1 (light grey) or MVATG18377 (dark grey). Results are shown as the number of IFNγ-producing T cells (spots-forming cells) per 10⁶ splenocytes following stimulation with either peptide pools specific of each of the 14 antigens or the irrelevant GLL peptide (Irr). For long sequence antigens (Rv2029, Rv2626, Rv1733, Rv0111, RpfB-RpfD, Ag85B, Rv3478 and Rv1807), only results obtained with the peptide pool leading to the highest response are shown. Full bars represent individual mice and hatched bars represent median values of each group. The experimental cut-off value (dotted line) is represented for each mouse strain: 51 spots/10⁶ cells for BALB/c, 56 spots/10⁶ cells for C57BL/6 and 72 spots/10⁶ cells for C3H/HeN mice. Results are representative of two independent experiments.</p

La tuberculose (TB), maladie pulmonaire causee par le Mycobacterium tuberculosis (Mtb), reste la ... more La tuberculose (TB), maladie pulmonaire causee par le Mycobacterium tuberculosis (Mtb), reste la premiere cause de mortalite par un agent infectieux. La TB est responsable de pres de 1,7 million de morts et de 10,4 millions de nouveaux cas par an dans le monde. L’emergence et la propagation de souches bacteriennes multi-resistantes aux antibiotiques (MDR) representent une menace majeure grandissante et refletent l’efficacite partielle des therapies actuelles. Le traitement des patients atteints de TB-MDR est constitue actuellement de combinaisons d’antibiotiques, souvent toxiques, administres pendant une longue duree, avec une efficacite limitee. Il existe donc un besoin urgent de developper de nouveaux traitements antituberculeux. L’immunotherapie, dont l’objectif est d’ameliorer la reponse immunitaire de l’hote contre le Mtb, represente une approche complementaire interessante dans le but de diminuer la duree et d’augmenter l’efficacite des traitements actuels de la TB-MDR. Le pre...

La presente invention concerne en general des combinaisons immunogenes comprenant au moins cinq a... more La presente invention concerne en general des combinaisons immunogenes comprenant au moins cinq antigenes d'une espece de Mycobacterium , ainsi qu'une fusion de ceux-ci, et des molecules d'acide nucleique codant pour de tels antigenes combines et une telle fusion. La presente invention concerne egalement des molecules d'acide nucleique, des vecteurs, des cellules hotes et des compositions comprenant ou codant pour lesdites combinaisons d'antigenes mycobacteriens et polypeptides de fusion, ainsi que des procedes pour la production recombinante de ceux-ci. La presente invention concerne egalement des procedes d'utilisation desdites combinaisons d'antigenes mycobacteriens, polypeptides de fusion, vecteurs, cellules hotes, compositions, en particulier pour l'induction ou la stimulation d'une reponse immunitaire dirigee contre une infection par Mycobacterium ou toute maladie provoquee par ou associee a une infection par Mycobacterium . La presente inve...

PloS one, 2018

Despite the existence of the prophylactic Bacille Calmette-Guérin (BCG) vaccine, infection by Myc... more Despite the existence of the prophylactic Bacille Calmette-Guérin (BCG) vaccine, infection by Mycobacterium tuberculosis (Mtb) remains a major public health issue causing up to 1.8 million annual deaths worldwide. Increasing prevalence of Mtb strains resistant to antibiotics represents an urgent threat for global health that has prompted a search for alternative treatment regimens not subject to development of resistance. Immunotherapy constitutes a promising approach to improving current antibiotic treatments through engagement of the host's immune system. We designed a multi-antigenic and multiphasic vaccine, based on the Modified Vaccinia Ankara (MVA) virus, denoted MVATG18598, which expresses ten antigens classically described as representative of each of different phases of Mtb infection. In vitro analysis coupled with multiple-passage evaluation demonstrated that this vaccine is genetically stable, i.e. fit for manufacturing. Using different mouse strains, we show that MVA...

PLOS ONE, 2015

Bacille Calmette-Guérin (BCG) vaccination of new born babies can protect children against tubercu... more Bacille Calmette-Guérin (BCG) vaccination of new born babies can protect children against tuberculosis (TB), but fails to protect adults consistently against pulmonary TB underlying the urgent need to develop novel TB vaccines. Majority of first generation TB vaccine candidates have relied on a very limited number of antigens typically belonging to the active phase of infection. We have designed a multi-antigenic and multiphasic vaccine, based on the Modified Vaccinia Ankara virus (MVA). Up to fourteen antigens representative of the three phases of TB infection (active, latent and resuscitation) were inserted into MVA. Using three different strains of mouse (BALB/c, C57BL/6 and C3H/HeN), we show that a single vaccination results in induction of both CD4 and CD8 T cells, displaying capacity to produce multiple cytokines together with cytolytic activity targeting a large array of epitopes. As expected, dominance of responses was linked to the mouse haplotype although for a given haplotype, responses specific of at least one antigen per phase could always be detected. Vaccination of non-human primates with the 14 antigens MVA-TB candidate resulted in broad and potent cellular-based immunogenicity. The remarkable plasticity of MVA opens the road to development of a novel class of highly complex recombinant TB vaccines to be evaluated in both prophylactic and therapeutic settings.

The Journal of Immunology, 2009

The Journal of Immunology, 2009

Most memory CD8 T cell subsets that have been hitherto defined are generated in response to infec... more Most memory CD8 T cell subsets that have been hitherto defined are generated in response to infectious pathogens. In this study, we have characterized the CD8 T cells that survive priming conditions, devoid of pathogen-derived danger signals. In both a TCR-transgenic model and a model of contact hypersensitivity, we show that the priming of naive CD8 T cells under sterile inflammatory conditions generates memory. The corresponding memory CD8 T cells can be identified by their intermediate expression levels of CD44 and CD122. We also show that CD44/122 int memory CD8 T cells spontaneously develop in wild type mice and that they display intermediate levels of several other memory traits including functional (IFN-␥ secretion capacity, CCL5 messenger stores), phenotypic, and molecular (T-bet and eomesodermin expression levels) features. We finally show that they correspond to an early differentiation stage and can further differentiate in CD44/122 high memory T cells. Altogether, our results identify a new memory CD8 T cell subset that is generated under sterile inflammatory conditions and involved in the recall contact hypersensitivity reactions that are responsible for allergic contact dermatitis.

The Journal of Immunology, 2006

The Journal of Immunology, 2012

IL-4 is one of the main cytokines produced during Th2-inducing pathologies. This cytokine has bee... more IL-4 is one of the main cytokines produced during Th2-inducing pathologies. This cytokine has been shown to affect a number of immune processes such as Th differentiation and innate immune responses. However, the impact of IL-4 on CD8 T cell responses remains unclear. In this study, we analyzed the effects of IL-4 on global gene expression profiles of Ag-induced memory CD8 T cells in the mouse. Gene ontology analysis of this signature revealed that IL-4 regulated most importantly genes associated with immune responses. Moreover, this IL-4 signature overlapped with the set of genes preferentially expressed by memory CD8 T cells over naive CD8 T cells. In particular, IL-4 downregulated in vitro and in vivo in a STAT6-dependent manner the memory-specific expression of NKG2D, thereby increasing the activation threshold of memory CD8 T cells. Furthermore, IL-4 impaired activation of memory cells as well as their differentiation into effector cells. This phenomenon could have an important clinical relevance as patients affected by Th2 pathologies such as parasitic infections or atopic dermatitis often suffer from viral-induced complications possibly linked to inefficient CD8 T cell responses.

Human Vaccines & Immunotherapeutics

<p>Groups of mice co-treated with antibiotics (ATB) and MVATG18598 or MVATGN33.1 given thri... more <p>Groups of mice co-treated with antibiotics (ATB) and MVATG18598 or MVATGN33.1 given thrice subcutaneously during antibiotic therapy were analyzed.</p

<p>Three antigen fusions were inserted in the deletion III of MVA vector. The first fusion ... more <p>Three antigen fusions were inserted in the deletion III of MVA vector. The first fusion is constituted by the fusion of Rv2029, Rv2626, Rv1733 and Rv0111 proteins and is placed under the control of p7.5K promoter. The second fusion contains a fusion of RpfB-RpfD, Ag85B, TB10.4 and ESAT-6 proteins and its expression is driven by the pH5R promoter. The third fusion is constituted by the fusion of Rv0569, Rv1813, Rv3407, Rv3478 and Rv1807 proteins and is placed under the control of pB2R promoter. SF, signal peptide of the F protein of measles virus. SR, signal peptide of the glycoprotein precursor of rabies virus ERA strain. TMR, membrane-anchoring peptide derived from the rabies glycoprotein of PG strain.</p

<p>Cells from MVATG18377-immunized BALB/c mice were stimulated with antigen peptide pools o... more <p>Cells from MVATG18377-immunized BALB/c mice were stimulated with antigen peptide pools or an irrelevant E7 peptide (Irr) and IFNγ, IL2 and TNFα intracellular cytokine staining was measured by flow cytometry. Results are presented as (<b>A</b>) the percentage of IFNγ⁺, IFNγ⁺TNFα⁺ or IFNγ⁺TNFα⁺IL2⁺ cell subsets among total CD4 T cells or (<b>B</b>) percentage of IFNγ⁺, TNFα⁺ or IFNγ⁺TNFα⁺ cell subsets among total CD8 T cells. Plain bars represent response from individual mice and hatched bars represent median response for each cell subset. Cut-off value (dotted line, 0.02%) is represented for both CD4⁺ and CD8⁺ T cell responses. Only antigens with median values above the cut-off value are represented. For these antigens, only cell subgroups with a percentage above the cut-off value are represented. No response was detected in MVATGN33.1-immunized mice (data not shown). For each cell population, background signal obtained in unstimulated cell condition was subtracted. Pie charts represent a more global analysis for each responder antigen. All analyzed single (IFNγ⁺, TNFα⁺ and IL2⁺), double (IFNγ⁺TNFα⁺, IFNγ⁺IL2⁺ and TNFα⁺IL2⁺) or triple (IFNγ⁺TNFα⁺IL2⁺) cytokine producer cells are included under the corresponding color codes. Results are representative of two independent experiments.</p

<p><b>(A)</b> Scheme of immunotherapy experiments. BALB/c mice were infected wi... more <p><b>(A)</b> Scheme of immunotherapy experiments. BALB/c mice were infected with a low dose aerosol of <i>M</i>. <i>tuberculosis</i> H37Rv. Four weeks later, mice were treated with an antibiotic regimen (RHZ) for 11 weeks. A subset of antibiotic-treated mice was vaccinated, through subcutaneous (SC) or intranasal (IN) routes, three times with a 3-week interval, with either MVATGN33.1 or MVATG18598 during (Weeks 7, 10 and 13) or after (Weeks 17, 20 and 23) chemotherapy. Twelve weeks after the end of the antibiotic regimen (Week 27), the number of viable bacteria in the lungs of animals was determined. <b>(B)</b> <i>M</i>. <i>tuberculosis</i> CFU counts after the completion of the antibiotic regimen in mice vaccinated or not (black) with either MVATGN33.1 (grey) or MVATG18598 (orange). Each symbol represents CFU value of individual mice. Dotted line indicates the lower limit of detection. Results are expressed as mean (black line) log₁₀ CFU ± S.E.M. Statistical analysis was performed using a Mann-Whitney test. *, <i>p</i><0.05. <b>(C)</b> Titers of antibodies specific to MVATG18598-expressed antigens at the relapse evaluation. Each symbol represents antibody titer (log₁₀) of individual mice vaccinated or not (black) with either MVATGN33.1 (grey) or MVATG18598 (orange). Black line represents median value for each group. Statistical analysis was performed using a Mann-Whitney test. §, <i>p</i><0.05 and §§, <i>p</i><0.01 for comparison with RHZ group. *, <i>p</i><0.05 and **, <i>p</i><0.01 for comparison between MVATGN33.1- and MVATG18598-vaccinated groups.</p

<p>Groups of mice co-treated with antibiotics (ATB) and MVATG18598 or MVATGN33.1 given thri... more <p>Groups of mice co-treated with antibiotics (ATB) and MVATG18598 or MVATGN33.1 given thrice subcutaneously during antibiotic therapy were analyzed.</p

La tuberculose (TB), maladie pulmonaire causée par le Mycobacterium tuberculosis (Mtb), reste la ... more La tuberculose (TB), maladie pulmonaire causée par le Mycobacterium tuberculosis (Mtb), reste la première cause de mortalité par un agent infectieux. La TB est responsable de près de 1,7 million de morts et de 10,4 millions de nouveaux cas par an dans le monde. L’émergence et la propagation de souches bactériennes multi-résistantes aux antibiotiques (MDR) représentent une menace majeure grandissante et reflètent l’efficacité partielle des thérapies actuelles. Le traitement des patients atteints de TB-MDR est constitué actuellement de combinaisons d’antibiotiques, souvent toxiques, administrés pendant une longue durée, avec une efficacité limitée. Il existe donc un besoin urgent de développer de nouveaux traitements antituberculeux. L’immunothérapie, dont l’objectif est d’améliorer la réponse immunitaire de l’hôte contre le Mtb, représente une approche complémentaire intéressante dans le but de diminuer la durée et d’augmenter l’efficacité des traitements actuels de la TB-MDR. Le pre...

<p>Mice were immunized twice with either MVATGN33.1 or MVATG18377. Labelled target cell pop... more <p>Mice were immunized twice with either MVATGN33.1 or MVATG18377. Labelled target cell populations pulsed with individual antigen peptide pool were adoptively transferred into those 7 days after last immunization. Results are expressed as mean ± S.D. of the percentage of specific killing based on the relative ratio of target cells present in MVATG18377-vaccinated mice compared to those in the MVATGN33.1-vaccinated mice. For each time point and antigen peptide pool (P), significant lysis is indicated by * (p<0.05) using a permutation resampling test.</p

<p>Cells from MVATG18377-immunized C57BL/6 mice were stimulated with antigen peptide pools ... more <p>Cells from MVATG18377-immunized C57BL/6 mice were stimulated with antigen peptide pools or an irrelevant E7 peptide (Irr). Results are presented as (<b>A</b>) the percentage of IFNγ⁺, IFNγ⁺TNFα⁺ or IFNγ⁺TNFα⁺IL2⁺ cell subsets among total CD4 T cells or (<b>B</b>) percentage of IFNγ⁺, TNFα⁺ or IFNγ⁺TNFα⁺ cell subsets among total CD8 T cells. Plain bars represent response from individual mice and hatched bars represent median response for each cell subset. Cut-off value (dotted line, 0.02%) is represented for both CD4⁺ and CD8⁺ T cell responses. Only antigens with median value above the cut-off value are represented. For these antigens, only cell subgroups with a percentage above the cut-off value are represented. No response was detected in MVATGN33.1-immunized mice (data not shown). For each cell population, background signal obtained in unstimulated cell condition was subtracted. Pie charts represent a more global analysis for each responder antigen. All analyzed single (IFNγ⁺, TNFα⁺ and IL2⁺), double (IFNγ⁺TNFα⁺, IFNγ⁺IL2⁺ and TNFα⁺IL2⁺) or triple (IFNγ⁺TNFα⁺IL2⁺) cytokine producer cells are included under the corresponding color codes. Results are representative of two independent experiments.</p

<p>Percentage of relapsing mice and bacterial burden in mice vaccinated SC with increasing ... more <p>Percentage of relapsing mice and bacterial burden in mice vaccinated SC with increasing vaccination number of MVATG18598 during antibiotic regimen.</p

<p><b>(A)</b> MVATG18598 contains the fusion Rv2626/T2a/Ag85B under the control... more <p><b>(A)</b> MVATG18598 contains the fusion Rv2626/T2a/Ag85B under the control of pB2R promoter, the fusion CFP10/ESAT6 under the control of pH5R promoter, the fusion TB10.4/Rv0287 under the control of pSE/L promoter, the fusion RpfB-RpfD under the control of p7.5K promoter and the fusion Rv3407/E2a/Rv1813 under the control of pA35R promoter. <b>(B)</b> CEF cells were infected or not (Mock) with MVATG18598 or MVATGN33.1 and cell extracts analyzed by Western blot. Fusion Rv2626/T2a/Ag85B was detected using a mouse monoclonal anti-Rv2626 antibody (expected molecular weight of cleaved product: 17.4 kDa) and a rabbit polyclonal anti-Ag85B antibody (expected molecular weight of cleaved product: 31.3 kDa). CFP10/ESAT6 fusion (expected molecular weight: 21.8 kDa) was detected using a mouse monoclonal anti-ESAT6 antibody. TB10.4/Rv0287 fusion (expected molecular weight: 21.2 kDa) was detected using an anti-TB10.4 rabbit polyclonal antibody. Fusion RpfB-RpfD (expected molecular weight: 37.0 kDa) was detected using a rabbit polyclonal anti-RpfB antibody. Fusion Rv3407/E2a/Rv1813 was detected using a rabbit polyclonal anti-Rv3407 antibody (expected molecular weight of cleaved product: 13.2 kDa) and a rabbit polyclonal anti-Rv1813 antibody (expected molecular weight of cleaved product: 12.1 kDa). ND; not done.</p

<p>(<b>A</b>) BALB/c, (<b>B</b>) C57BL/6 and (<b>C</b>)... more <p>(<b>A</b>) BALB/c, (<b>B</b>) C57BL/6 and (<b>C</b>) C3H/HeN mice were immunized once with either MVATGN33.1 (light grey) or MVATG18377 (dark grey). Results are shown as the number of IFNγ-producing T cells (spots-forming cells) per 10⁶ splenocytes following stimulation with either peptide pools specific of each of the 14 antigens or the irrelevant GLL peptide (Irr). For long sequence antigens (Rv2029, Rv2626, Rv1733, Rv0111, RpfB-RpfD, Ag85B, Rv3478 and Rv1807), only results obtained with the peptide pool leading to the highest response are shown. Full bars represent individual mice and hatched bars represent median values of each group. The experimental cut-off value (dotted line) is represented for each mouse strain: 51 spots/10⁶ cells for BALB/c, 56 spots/10⁶ cells for C57BL/6 and 72 spots/10⁶ cells for C3H/HeN mice. Results are representative of two independent experiments.</p

La tuberculose (TB), maladie pulmonaire causee par le Mycobacterium tuberculosis (Mtb), reste la ... more La tuberculose (TB), maladie pulmonaire causee par le Mycobacterium tuberculosis (Mtb), reste la premiere cause de mortalite par un agent infectieux. La TB est responsable de pres de 1,7 million de morts et de 10,4 millions de nouveaux cas par an dans le monde. L’emergence et la propagation de souches bacteriennes multi-resistantes aux antibiotiques (MDR) representent une menace majeure grandissante et refletent l’efficacite partielle des therapies actuelles. Le traitement des patients atteints de TB-MDR est constitue actuellement de combinaisons d’antibiotiques, souvent toxiques, administres pendant une longue duree, avec une efficacite limitee. Il existe donc un besoin urgent de developper de nouveaux traitements antituberculeux. L’immunotherapie, dont l’objectif est d’ameliorer la reponse immunitaire de l’hote contre le Mtb, represente une approche complementaire interessante dans le but de diminuer la duree et d’augmenter l’efficacite des traitements actuels de la TB-MDR. Le pre...

La presente invention concerne en general des combinaisons immunogenes comprenant au moins cinq a... more La presente invention concerne en general des combinaisons immunogenes comprenant au moins cinq antigenes d'une espece de Mycobacterium , ainsi qu'une fusion de ceux-ci, et des molecules d'acide nucleique codant pour de tels antigenes combines et une telle fusion. La presente invention concerne egalement des molecules d'acide nucleique, des vecteurs, des cellules hotes et des compositions comprenant ou codant pour lesdites combinaisons d'antigenes mycobacteriens et polypeptides de fusion, ainsi que des procedes pour la production recombinante de ceux-ci. La presente invention concerne egalement des procedes d'utilisation desdites combinaisons d'antigenes mycobacteriens, polypeptides de fusion, vecteurs, cellules hotes, compositions, en particulier pour l'induction ou la stimulation d'une reponse immunitaire dirigee contre une infection par Mycobacterium ou toute maladie provoquee par ou associee a une infection par Mycobacterium . La presente inve...

PloS one, 2018

Despite the existence of the prophylactic Bacille Calmette-Guérin (BCG) vaccine, infection by Myc... more Despite the existence of the prophylactic Bacille Calmette-Guérin (BCG) vaccine, infection by Mycobacterium tuberculosis (Mtb) remains a major public health issue causing up to 1.8 million annual deaths worldwide. Increasing prevalence of Mtb strains resistant to antibiotics represents an urgent threat for global health that has prompted a search for alternative treatment regimens not subject to development of resistance. Immunotherapy constitutes a promising approach to improving current antibiotic treatments through engagement of the host's immune system. We designed a multi-antigenic and multiphasic vaccine, based on the Modified Vaccinia Ankara (MVA) virus, denoted MVATG18598, which expresses ten antigens classically described as representative of each of different phases of Mtb infection. In vitro analysis coupled with multiple-passage evaluation demonstrated that this vaccine is genetically stable, i.e. fit for manufacturing. Using different mouse strains, we show that MVA...

PLOS ONE, 2015

Bacille Calmette-Guérin (BCG) vaccination of new born babies can protect children against tubercu... more Bacille Calmette-Guérin (BCG) vaccination of new born babies can protect children against tuberculosis (TB), but fails to protect adults consistently against pulmonary TB underlying the urgent need to develop novel TB vaccines. Majority of first generation TB vaccine candidates have relied on a very limited number of antigens typically belonging to the active phase of infection. We have designed a multi-antigenic and multiphasic vaccine, based on the Modified Vaccinia Ankara virus (MVA). Up to fourteen antigens representative of the three phases of TB infection (active, latent and resuscitation) were inserted into MVA. Using three different strains of mouse (BALB/c, C57BL/6 and C3H/HeN), we show that a single vaccination results in induction of both CD4 and CD8 T cells, displaying capacity to produce multiple cytokines together with cytolytic activity targeting a large array of epitopes. As expected, dominance of responses was linked to the mouse haplotype although for a given haplotype, responses specific of at least one antigen per phase could always be detected. Vaccination of non-human primates with the 14 antigens MVA-TB candidate resulted in broad and potent cellular-based immunogenicity. The remarkable plasticity of MVA opens the road to development of a novel class of highly complex recombinant TB vaccines to be evaluated in both prophylactic and therapeutic settings.

The Journal of Immunology, 2009

The Journal of Immunology, 2009

Most memory CD8 T cell subsets that have been hitherto defined are generated in response to infec... more Most memory CD8 T cell subsets that have been hitherto defined are generated in response to infectious pathogens. In this study, we have characterized the CD8 T cells that survive priming conditions, devoid of pathogen-derived danger signals. In both a TCR-transgenic model and a model of contact hypersensitivity, we show that the priming of naive CD8 T cells under sterile inflammatory conditions generates memory. The corresponding memory CD8 T cells can be identified by their intermediate expression levels of CD44 and CD122. We also show that CD44/122 int memory CD8 T cells spontaneously develop in wild type mice and that they display intermediate levels of several other memory traits including functional (IFN-␥ secretion capacity, CCL5 messenger stores), phenotypic, and molecular (T-bet and eomesodermin expression levels) features. We finally show that they correspond to an early differentiation stage and can further differentiate in CD44/122 high memory T cells. Altogether, our results identify a new memory CD8 T cell subset that is generated under sterile inflammatory conditions and involved in the recall contact hypersensitivity reactions that are responsible for allergic contact dermatitis.

The Journal of Immunology, 2006

The Journal of Immunology, 2012

IL-4 is one of the main cytokines produced during Th2-inducing pathologies. This cytokine has bee... more IL-4 is one of the main cytokines produced during Th2-inducing pathologies. This cytokine has been shown to affect a number of immune processes such as Th differentiation and innate immune responses. However, the impact of IL-4 on CD8 T cell responses remains unclear. In this study, we analyzed the effects of IL-4 on global gene expression profiles of Ag-induced memory CD8 T cells in the mouse. Gene ontology analysis of this signature revealed that IL-4 regulated most importantly genes associated with immune responses. Moreover, this IL-4 signature overlapped with the set of genes preferentially expressed by memory CD8 T cells over naive CD8 T cells. In particular, IL-4 downregulated in vitro and in vivo in a STAT6-dependent manner the memory-specific expression of NKG2D, thereby increasing the activation threshold of memory CD8 T cells. Furthermore, IL-4 impaired activation of memory cells as well as their differentiation into effector cells. This phenomenon could have an important clinical relevance as patients affected by Th2 pathologies such as parasitic infections or atopic dermatitis often suffer from viral-induced complications possibly linked to inefficient CD8 T cell responses.