213 results on '"Bonnefoy JY"'
Search Results
2. Structural basis for SH2D1A mutations in X-linked lymphoproliferative disease
- Author
-
Giliani, Silvia Clara, Lappalainen, I, Franceschini, R, Bonnefoy, Jy, Duckett, C, Notarangelo, Luigi Daniele, and Vihinen, M.
- Published
- 2000
3. CD23/FC epsilon RII is constitutively expressed on human intestinal epithelium, and upregulated in cow's milk protein intolerance
- Author
-
Kaiserlian, D., Lachaux, A., Grosjean, I., Graber, P., Bonnefoy, Jy, and Deleage, Gilbert
- Subjects
[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology - Abstract
xxx
- Published
- 1995
4. Intestinal epithelial cells express the CD23/Fc epsilon RII molecule: enhanced expression in enteropathies
- Author
-
Kaiserlian, D., Lachaux, A., Grosjean, I., Graber, P., Bonnefoy, Jy, and Deleage, Gilbert
- Subjects
Receptors, IgE ,Blotting, Western ,Antibodies, Monoclonal ,Fluorescent Antibody Technique ,Epithelial Cells ,Flow Cytometry ,Immunohistochemistry ,Epithelium ,Intestines ,Intestinal Diseases ,hemic and lymphatic diseases ,[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Humans ,Research Article - Abstract
Immunohistochemical analysis of normal human intestine revealed that two anti-CD23 monoclonal antibodies (mAb), EBVCS 1 and EBVCS 2, reacted with human intestinal epithelial cells. Both mAb exhibited an exclusive reactivity with epithelial cells of the small and large bowels. Staining with both EBVCS 1 and EBVCS 2 was localized on the apical and basal sides of enterocytes. Enhanced expression of CD23 on gut epithelial cells was found in inflammatory bowel diseases, in children with food intolerance to cows' milk proteins and in a young infant with severe autoimmune enteropathy. Western blot analysis of anti-CD23 mAb reactivity with gut epithelial cell extracts showed the presence of a non-reducible 42,000-45,000 M(r) polypeptide compatible with the membrane form of the intact CD23 molecule. These data show that CD23 is constitutively expressed by intestinal epithelial cells and that its expression is enhanced in enteropathies.Immunohistochemical analysis of normal human intestine revealed that two anti-CD23 monoclonal antibodies (mAb), EBVCS 1 and EBVCS 2, reacted with human intestinal epithelial cells. Both mAb exhibited an exclusive reactivity with epithelial cells of the small and large bowels. Staining with both EBVCS 1 and EBVCS 2 was localized on the apical and basal sides of enterocytes. Enhanced expression of CD23 on gut epithelial cells was found in inflammatory bowel diseases, in children with food intolerance to cows' milk proteins and in a young infant with severe autoimmune enteropathy. Western blot analysis of anti-CD23 mAb reactivity with gut epithelial cell extracts showed the presence of a non-reducible 42,000-45,000 M(r) polypeptide compatible with the membrane form of the intact CD23 molecule. These data show that CD23 is constitutively expressed by intestinal epithelial cells and that its expression is enhanced in enteropathies.
- Published
- 1993
5. EFFECTS OF SERUM ON LECTIN-INDUCED LYMPHOCYTE PROLIFERATION
- Author
-
P. W. Ledger, Bonnefoy Jy, A. Réano, and J. Thivollet
- Subjects
biology ,Chemistry ,biology.protein ,Lectin ,Lymphocyte proliferation ,Molecular biology - Published
- 1984
6. Manno-glycoconjugates of human keratinocytes in culture: effect of retinoic acid
- Author
-
Bonnefoy Jy, P. W. Ledger, Alain Reano, and Jean Thivolet
- Subjects
chemistry.chemical_classification ,Dose-Response Relationship, Drug ,Chemistry ,Glycoconjugate ,Retinoic acid ,Retinoic acid receptor beta ,Tretinoin ,Dermatology ,Retinoic acid receptor ,chemistry.chemical_compound ,Biochemistry ,Humans ,Electrophoresis, Polyacrylamide Gel ,Mannose ,Cells, Cultured ,Glycoproteins ,Skin - Published
- 1984
7. Prospective Randomized Study of the St. Jude Medical®, BjÖrk-Shiley®, and Starr-Edwards® 6120 Valve Prostheses in the Mitral Position
- Author
-
M. Ferrini, Mikaeloff P, Coll-Mazzei J, Jegaden O, Rumolo A, and Bonnefoy Jy
- Subjects
medicine.medical_specialty ,business.industry ,medicine.medical_treatment ,Mitral valve replacement ,Valve prosthesis ,Hemodynamics ,medicine.disease ,Prosthesis ,Group B ,Surgery ,Aortic valve replacement ,Valve replacement ,medicine ,Prospective randomized study ,business - Abstract
During a 5-year period (January 1979 to December 1983), 357 patients received mitral valve replacement. Group A was comprised of 179 patients receiving a ST. JUDE MEDICAL® prosthesis in the mitral position. Group B included 178 patients; initially 113 patients were implanted with a BJORK-SHILEY® valve, and later 65 patients received a STARR-EDWARDS® 6120 valve prosthesis in the mitral position. All implants were performed by the same surgeon and the groups were randomized. Analysis of 21 preoperative (clinical and hemodynamic data) and operative variables showed the groups to be well randomized. All patients were anticoagulated postoperatively. A follow-up study was performed each year postoperatively. At the end of 1986, there was a 35-month to 95-month follow-up with a mean of 64.7 months (1596 patient-years of follow-up). Fifteen patients were lost to follow-up. The actuarial survival rate is significantly different (p < 0.05) at 5 years with 87.6 ± 4.5% for Group A vs. 77.4 ± 6% for Group B, and at 7 years with 83.4 ± 6.5% for Group A vs. 73.2 ± 7.2% for Group B. In conclusion, in the mitral position, the ST. JUDE MEDICAL prosthesis gives a significant benefit compared to BJORK-SHILEY or STARR-EDWARDS 6120 prostheses.
- Published
- 1989
8. Concanavalin (ConA)-reactive human epidermal glycoproteins
- Author
-
Bonnefoy, JY, primary, Reano, A, additional, Ledger, PW, additional, and Thivolet, J., additional
- Published
- 1984
- Full Text
- View/download PDF
9. Hepatitis C vaccine: supply and demand.
- Author
-
Inchauspé G, Honnet G, Bonnefoy JY, Nicosia A, and Strickland GT
- Published
- 2008
- Full Text
- View/download PDF
10. Therapeutic vaccination with TG4010 and first-line chemotherapy in advanced non-small-cell lung cancer: a controlled phase 2B trial.
- Author
-
Quoix E, Ramlau R, Westeel V, Papai Z, Madroszyk A, Riviere A, Koralewski P, Breton JL, Stoelben E, Braun D, Debieuvre D, Lena H, Buyse M, Chenard MP, Acres B, Lacoste G, Bastien B, Tavernaro A, Bizouarne N, and Bonnefoy JY
- Abstract
BACKGROUND: Chemotherapy is the standard of care for advanced stages of non-small-cell lung cancer (NSCLC). TG4010 is a targeted immunotherapy based on a poxvirus (modified vaccinia virus Ankara) that codes for MUC1 tumour-associated antigen and interleukin 2. This study assessed TG4010 in combination with first-line chemotherapy in advanced NSCLC. METHODS: 148 patients with advanced (stage IIIB [wet] or IV) NSCLC expressing MUC1 by immunohistochemistry, and with performance status 0 or 1, were enrolled in parallel groups in this open-label, phase 2B study. 74 patients were allocated to the combination therapy group, and received TG4010 (10(8) plaque forming units) plus cisplatin (75 mg/m(2) on day 1) and gemcitabine (1250 mg/m(2) on days 1 and 8) repeated every 3 weeks for up to six cycles. 74 patients allocated to the control group received the same chemotherapy alone. Patients were allocated using a dynamic minimisation procedure stratified by centre, performance status, and disease stage. The primary endpoint was 6-month progression-free survival (PFS), with a target rate of 40% or higher in the experimental group. Analyses were done on an intention-to-treat basis. This study is completed and is registered with ClinicalTrials.gov, number NCT00415818. FINDINGS: 6-month PFS was 43·2% (32 of 74; 95% CI 33·4-53·5) in the TG4010 plus chemotherapy group, and 35·1% (26 of 74; 25·9-45·3) in the chemotherapy alone group. Fever, abdominal pain, and injection-site pain of any grade according to National Cancer Institute Common Toxicity Criteria were more common in the TG4010 group than in the chemotherapy alone group: 17 of 73 patients (23·3%) versus six of 72 (8·3%), 12 (16·4%) versus two (2·8%), and four (5·5%) versus zero (0%), respectively. The most common grade 3-4 adverse events were neutropenia (33 [45·2%] of patients in the TG4010 plus chemotherapy group vs 31 [43·1%] in the chemotherapy alone group) and fatigue (18 [24·7%] vs 13 [18·1%]); the only grade 3-4 events that differed significantly between groups were anorexia (three [4·1%] vs 10 [13·9%]) and pleural effusion (none vs four [5·6%]). 38 of 73 patients (52·1%) in the TG4010 plus chemotherapy group and 34 of 72 (47·2%) in the chemotherapy alone group had at least one serious adverse event. INTERPRETATION: This phase 2B study suggests that TG4010 enhances the effect of chemotherapy in advanced NSCLC. A confirmatory phase 2B-3 trial has been initiated. FUNDING: Transgene SA, Advanced Diagnostics for New Therapeutic Approaches (ADNA)/OSEO. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
11. MAPN: First-in-Class Reagent for Kinetically Resolved Thiol-to-Thiol Conjugation.
- Author
-
Koniev O, Kolodych S, Baatarkhuu Z, Stojko J, Eberova J, Bonnefoy JY, Cianférani S, Van Dorsselaer A, and Wagner A
- Subjects
- Cell Line, Tumor, Humans, Immunoconjugates chemistry, Indicators and Reagents chemistry, Kinetics, Alkynes chemistry, Maleimides chemistry, Sulfhydryl Compounds chemistry
- Abstract
Thiols are among the most frequently used functional groups in the field of bioconjugation. While there exists a variety of heterobifunctional reagents that allow for coupling thiols to other functions (e.g., amines, carboxylic acids), there is no specific reagent for creating heteroconjugates using two different thiols. In response to the ever-increasing demand for bioconjugation tools, we have developed p-(maleimide)-phenylpropionitrile (MAPN)-an efficient reagent for kinetically resolved thiol-to-thiol coupling. In a comparative study with its closest commercially available analogue, p-phenylenedimaleimide, MAPN has shown substantial advantages for the preparation of thiol-thiol heteroconjugates. Namely, an antibody-drug conjugate (ADC) with mertansine (DM1), conjugated to the cysteine residues of Trastuzumab, was prepared for the first time.
- Published
- 2015
- Full Text
- View/download PDF
12. CBTF: new amine-to-thiol coupling reagent for preparation of antibody conjugates with increased plasma stability.
- Author
-
Kolodych S, Koniev O, Baatarkhuu Z, Bonnefoy JY, Debaene F, Cianférani S, Van Dorsselaer A, and Wagner A
- Subjects
- Cell Line, Halogenation, Humans, Immunoconjugates blood, Maleimides chemistry, Amines chemistry, Benzene Derivatives chemistry, Cross-Linking Reagents chemistry, Immunoconjugates chemistry, Sulfhydryl Compounds chemistry
- Abstract
Amine-to-thiol coupling is the most common route for the preparation of antibody-drug conjugates (ADC). It is usually achieved by using heterobifunctional reagents possessing an activated ester at one end and a maleimide group at the other. However, maleimide-based conjugates were recently revealed to have limited stability in blood circulation, which can compromise therapeutic efficacy of the conjugate. To address this issue, we have developed a heterobifunctional reagent, sodium 4-((4-(cyanoethynyl)benzoyl)oxy)-2,3,5,6-tetrafluorobenzenesulfonate (CBTF), for amine-to-thiol coupling. It comprises a recently described 3-arylpropionitrile (APN) function in replacement of maleimide and allows for the preparation of remarkably stable conjugates. A series of antibody-dye conjugates have been prepared using this reagent and shown superior stability in human blood plasma compared to maleimide-derived conjugates.
- Published
- 2015
- Full Text
- View/download PDF
13. Bone- and cartilage-protective effects of a monoclonal antibody against colony-stimulating factor 1 receptor in experimental arthritis.
- Author
-
Toh ML, Bonnefoy JY, Accart N, Cochin S, Pohle S, Haegel H, De Meyer M, Zemmour C, Preville X, Guillen C, Thioudellet C, Ancian P, Lux A, Sehnert B, Nimmerjahn F, Voll RE, and Schett G
- Subjects
- Animals, Antibodies, Monoclonal immunology, Antibodies, Monoclonal therapeutic use, Arthritis, Experimental drug therapy, Arthritis, Experimental metabolism, Arthritis, Rheumatoid metabolism, Bone and Bones drug effects, Bone and Bones metabolism, Cartilage drug effects, Cartilage metabolism, Case-Control Studies, Dendritic Cells metabolism, Dendritic Cells pathology, Disease Models, Animal, Female, Humans, Macrophages metabolism, Macrophages pathology, Male, Mice, Mice, Inbred DBA, Middle Aged, Monocytes metabolism, Monocytes pathology, Osteoarthritis metabolism, Osteoclasts metabolism, Osteoclasts pathology, Receptor, Macrophage Colony-Stimulating Factor drug effects, Receptor, Macrophage Colony-Stimulating Factor metabolism, Synovial Membrane drug effects, Synovial Membrane metabolism, Synovial Membrane pathology, Antibodies, Monoclonal pharmacology, Arthritis, Experimental pathology, Arthritis, Rheumatoid pathology, Bone and Bones pathology, Cartilage pathology, Osteoarthritis pathology, Receptor, Macrophage Colony-Stimulating Factor antagonists & inhibitors
- Abstract
Objective: Colony-stimulating factor 1 receptor (CSF-1R) essentially modulates monocyte proliferation, migration, and activation, which are considered important for the pathogenesis of rheumatoid arthritis (RA). We undertook this study to determine CSF-1R expression in human RA as well as the efficacy of a specific anti-CSF-1R monoclonal antibody (AFS98) in 2 different animal models of RA., Methods: CSF-1R expression was examined in blood, synovium, and bone samples from RA patients, osteoarthritis (OA) patients, and healthy subjects. The efficacy of AFS98 was examined by clinical assessment, histology, and bone histomorphometry in collagen-induced arthritis (CIA) and serum-transfer arthritis., Results: CSF-1R expression was increased in the synovium of RA patients compared to OA patients and healthy controls in fibroblast-like synoviocytes, follicular dendritic cells, macrophages, and osteoclasts. Circulating RA monocytes and neutrophils but not lymphocytes were CSF-1R+. In mice, blockade of CSF-1R abrogated cartilage damage, bone erosion, and systemic bone loss, and this was associated with the depletion of osteoclasts in both models. While blockade of CSF-1R did not affect inflammation in passive serum-transfer arthritis, it significantly reduced inflammation in CIA, and this was associated with the absence of synovial macrophages and reduced splenic CD11b+Gr-1- monocytes., Conclusion: CSF-1R was broadly expressed in human RA. Blockade of CSF-1R protected against bone and cartilage destruction in both mouse models and also showed significant antiinflammatory effects in the CIA model. These data provide evidence for CSF-1R as a therapeutic target in RA., (Copyright © 2014 by the American College of Rheumatology.)
- Published
- 2014
- Full Text
- View/download PDF
14. Yeast virus-derived stimulator of the innate immune system augments the efficacy of virus vector-based immunotherapy.
- Author
-
Claudepierre MC, Hortelano J, Schaedler E, Kleinpeter P, Geist M, Remy-Ziller C, Brandely R, Tosch C, Laruelle L, Jawhari A, Menguy T, Marchand JB, Romby P, Schultz P, Hartmann G, Rooke R, Bonnefoy JY, Preville X, and Rittner K
- Subjects
- Animals, Cell Line, Cytokines metabolism, Disease Models, Animal, Immunologic Factors isolation & purification, Immunologic Factors therapeutic use, Immunotherapy methods, Mice, Mice, Inbred C57BL, RNA, Double-Stranded isolation & purification, RNA, Double-Stranded therapeutic use, RNA, Viral isolation & purification, RNA, Viral therapeutic use, Survival Analysis, Dendritic Cells immunology, Immunologic Factors immunology, Neoplasms therapy, RNA, Double-Stranded immunology, RNA, Viral immunology, Saccharomyces cerevisiae virology, Toll-Like Receptor 3 immunology
- Abstract
Unlabelled: To identify novel stimulators of the innate immune system, we constructed a panel of eight HEK293 cell lines double positive for human Toll-like receptors (TLRs) and an NF-κB-inducible reporter gene. Screening of a large variety of compounds and cellular extracts detected a TLR3-activating compound in a microsomal yeast extract. Fractionation of this extract identified an RNA molecule of 4.6 kb, named nucleic acid band 2 (NAB2), that was sufficient to confer the activation of TLR3. Digests with single- and double-strand-specific RNases showed the double-strand nature of this RNA, and its sequence was found to be identical to that of the genome of the double-stranded RNA (dsRNA) L-BC virus of Saccharomyces cerevisiae. A large-scale process of production and purification of this RNA was established on the basis of chemical cell lysis and dsRNA-specific chromatography. NAB2 complexed with the cationic lipid Lipofectin but neither NAB2 nor Lipofectin alone induced the secretion of interleukin-12(p70) [IL-12(p70)], alpha interferon, gamma interferon-induced protein 10, macrophage inflammatory protein 1β, or IL-6 in human monocyte-derived dendritic cells. While NAB2 activated TLR3, Lipofectin-stabilized NAB2 also signaled via the cytoplasmic sensor for RNA recognition MDA-5. A significant increase of RMA-MUC1 tumor rejection and survival was observed in C57BL/6 mice after prophylactic vaccination with MUC1-encoding modified vaccinia virus Ankara (MVA) and NAB2-Lipofectin. This combination of immunotherapies strongly increased at the injection sites the percentage of infiltrating natural killer (NK) cells and plasmacytoid dendritic cells (pDCs), cell types which can modulate innate and adaptive immune responses., Importance: Virus-based cancer vaccines offer a good alternative to the treatment of cancer but could be improved. Starting from a screening approach, we have identified and characterized an unexplored biological molecule with immunomodulatory characteristics which augments the efficacy of an MVA-based immunotherapeutic agent. The immune modulator consists of the purified dsRNA genome isolated from a commercially used yeast strain, NAB2, mixed with a cationic lipid, Lipofectin. NAB2-Lipofectin stimulates the immune system via TLR3 and MDA-5. When it was injected at the MVA vaccination site, the immune modulator increased survival in a preclinical tumor model. We could demonstrate that NAB2-Lipofectin augments the MVA-induced infiltration of natural killer and plasmacytoid dendritic cells. We suggest indirect mechanisms of activation of these cell types by the influence of NAB2-Lipofectin on innate and adaptive immunity. Detailed analysis of cell migration at the vaccine injection site and the appropriate choice of an immune modulator should be considered to achieve the rational improvement of virus vector-based vaccination by immune modulators.
- Published
- 2014
- Full Text
- View/download PDF
15. 3D modeling and characterization of the human CD115 monoclonal antibody H27K15 epitope and design of a chimeric CD115 target.
- Author
-
Grellier B, Le Pogam F, Vitorino M, Starck JP, Geist M, Duong V, Haegel H, Menguy T, Bonnefoy JY, Marchand JB, and Ancian P
- Subjects
- Animals, Antibodies, Monoclonal chemistry, Antibodies, Monoclonal immunology, Computational Biology, Enzyme-Linked Immunosorbent Assay, Epitopes chemistry, Humans, Immunoglobulin Variable Region chemistry, Macrophage Colony-Stimulating Factor immunology, Mice, Models, Chemical, Protein Binding, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins immunology, Sequence Alignment, Antibodies, Monoclonal metabolism, Interleukins immunology, Macrophage Colony-Stimulating Factor metabolism, Receptor, Macrophage Colony-Stimulating Factor immunology, Recombinant Fusion Proteins metabolism
- Abstract
The humanized monoclonal antibody H27K15 specifically targets human CD115, a type III tyrosine kinase receptor involved in multiple cancers and inflammatory diseases. Binding of H27K15 to hCD115 expressing cells inhibits the functional effect of colony-stimulating factor-1 (CSF-1), in a non-competitive manner. Both homology modeling and docking programs were used here to model the human CD115 extracellular domains, the H27K15 variable region and their interaction. The resulting predicted H27K15 epitope includes mainly the D1 domain in the N-terminal extracellular region of CD115 and some residues of the D2 domain. Sequence alignment with the non-binding murine CD115, enzyme-linked immunosorbent assay, nuclear magnetic resonance spectroscopy and affinity measurements by quartz crystal microbalance revealed critical residues of this epitope that are essential for H27K15 binding. A combination of computational simulations and biochemical experiments led to the design of a chimeric CD115 carrying the human epitope of H27K15 in a murine CD115 backbone that is able to bind both H27K15 as well as the murine ligands CSF-1 and IL-34. These results provide new possibilities to minutely study the functional effects of H27K15 in a transgenic mouse that would express this chimeric molecule.
- Published
- 2014
- Full Text
- View/download PDF
16. Immunological characterization of a modified vaccinia virus Ankara vector expressing the human papillomavirus 16 E1 protein.
- Author
-
Remy-Ziller C, Germain C, Spindler A, Hoffmann C, Silvestre N, Rooke R, Bonnefoy JY, and Préville X
- Subjects
- Animals, Female, Humans, Mice, Mice, Inbred C57BL, Oncogene Proteins, Viral genetics, Papillomavirus Vaccines administration & dosage, Papillomavirus Vaccines genetics, Vaccines, Synthetic administration & dosage, Vaccines, Synthetic genetics, Vaccines, Synthetic immunology, CD8-Positive T-Lymphocytes immunology, Drug Carriers, Immunity, Cellular, Oncogene Proteins, Viral immunology, Papillomavirus Vaccines immunology, T-Lymphocytes, Cytotoxic immunology, Vaccinia virus genetics
- Abstract
Women showing normal cytology but diagnosed with a persistent high-risk human papillomavirus (HR-HPV) infection have a higher risk of developing high-grade cervical intraepithelial neoplasia and cervical cancer than noninfected women. As no therapeutic management other than surveillance is offered to these women, there is a major challenge to develop novel targeted therapies dedicated to the treatment of these patients. As such, E1 and E2 antigens, expressed early in the HPV life cycle, represent very interesting candidates. Both proteins are necessary for maintaining coordinated viral replication and gene synthesis during the differentiation process of the epithelium and are essential for the virus to complete its normal and propagative replication cycle. In the present study, we evaluated a new active targeted immunotherapeutic, a modified vaccinia virus Ankara (MVA) vector containing the E1 sequence of HPV16, aimed at inducing cellular immune responses with the potential to help and clear persistent HPV16-related infection. We carried out an extensive comparative time course analysis of the cellular immune responses induced by different schedules of immunization in C57BL/6 mice. We showed that multiple injections of MVA-E1 allowed sustained HPV16 E1-specific cellular immune responses in vaccinated mice and had no impact on the exhaustion phenotype of the generated HPV16 E1-specific CD8⁺ T cells, but they led to the differentiation of multifunctional effector T cells with high cytotoxic capacity. This study provides proof of concept that an MVA expressing HPV16 E1 can induce robust and long-lasting E1-specific responses and warrants further development of this candidate.
- Published
- 2014
- Full Text
- View/download PDF
17. Lymphocytic infiltration in the cutaneous lymphoma microenvironment after injection of TG1042.
- Author
-
Accart N, Urosevic-Maiwald M, Dummer R, Bataille V, Kehrer N, Niculescu C, Limacher JM, Chenard MP, Bonnefoy JY, and Rooke R
- Subjects
- Disease Progression, Humans, Lymphoma, T-Cell, Cutaneous pathology, Treatment Outcome, Genetic Vectors administration & dosage, Lymphocytes, Tumor-Infiltrating immunology, Lymphoma, T-Cell, Cutaneous immunology, Lymphoma, T-Cell, Cutaneous therapy, Tumor Microenvironment immunology
- Abstract
Background: Primary cutaneous lymphomas (CLs), characterized by an accumulation of clonal T or B lymphocytes preferentially localized in the skin, have been successfully treated with interferons (IFNs) which counterbalance the Th2-immunosuppressive state associated with this pathology. In a phase I/II clinical trial, we correlated the local immune infiltrate and the anti-tumor effects of repeated intralesional administrations of an adenovirus vector expressing human interferon-gamma (IFN-g) termed TG1042, in patients with advanced primary cutaneous T-cell lymphomas (CTCL) or multilesional cutaneous B-cell lymphomas (CBCL)., Methods: For each patient, variation in time of specific lymphocyte populations, defined by immunohistochemical stainings, was assessed in biopsies of injected lesions. For each patient, the change in local immune response was associated with the patient's objective response at the end of the study., Results: Immunohistochemical analyses of biopsies indicate that infiltration of CD8+ T lymphocytes and of TIA-1+ cytotoxic T-cells in lesions injected with TG1042 correlates with clinical benefit., Conclusions: These data suggest for the first time that a CD8+ cytotoxic infiltrate, induced by local expression of IFN-g correlates with a clinical response., Trial Registration: The phase I step (TG1042.01) does not have a registration number. The phase II step (TG1042.06) registration number was NCT00394693.
- Published
- 2013
- Full Text
- View/download PDF
18. Therapeutic effects of anti-CD115 monoclonal antibody in mouse cancer models through dual inhibition of tumor-associated macrophages and osteoclasts.
- Author
-
Fend L, Accart N, Kintz J, Cochin S, Reymann C, Le Pogam F, Marchand JB, Menguy T, Slos P, Rooke R, Fournel S, Bonnefoy JY, Préville X, and Haegel H
- Subjects
- Animals, Antibodies, Monoclonal immunology, Bone Neoplasms secondary, Breast Neoplasms pathology, Cell Line, Tumor, Disease Models, Animal, Female, Heterografts, Humans, Mice, Mice, Inbred C57BL, Mice, Nude, Survival Analysis, Antibodies, Monoclonal therapeutic use, Macrophages immunology, Neoplasms, Experimental therapy, Osteoclasts immunology, Receptor, Macrophage Colony-Stimulating Factor immunology
- Abstract
Tumor progression is promoted by Tumor-Associated Macrophages (TAMs) and metastasis-induced bone destruction by osteoclasts. Both myeloid cell types depend on the CD115-CSF-1 pathway for their differentiation and function. We used 3 different mouse cancer models to study the effects of targeting cancer host myeloid cells with a monoclonal antibody (mAb) capable of blocking CSF-1 binding to murine CD115. In mice bearing sub-cutaneous EL4 tumors, which are CD115-negative, the anti-CD115 mAb depleted F4/80(+) CD163(+) M2-type TAMs and reduced tumor growth, resulting in prolonged survival. In the MMTV-PyMT mouse model, the spontaneous appearance of palpable mammary tumors was delayed when the anti-CD115 mAb was administered before malignant transition and tumors became palpable only after termination of the immunotherapy. When administered to mice already bearing established PyMT tumors, anti-CD115 treatment prolonged their survival and potentiated the effect of chemotherapy with Paclitaxel. As shown by immunohistochemistry, this therapeutic effect correlated with the depletion of F4/80(+)CD163(+) M2-polarized TAMs. In a breast cancer model of bone metastasis, the anti-CD115 mAb potently blocked the differentiation of osteoclasts and their bone destruction activity. This resulted in the inhibition of cancer-induced weight loss. CD115 thus represents a promising target for cancer immunotherapy, since a specific blocking antibody may not only inhibit the growth of a primary tumor through TAM depletion, but also metastasis-induced bone destruction through osteoclast inhibition.
- Published
- 2013
- Full Text
- View/download PDF
19. A unique anti-CD115 monoclonal antibody which inhibits osteolysis and skews human monocyte differentiation from M2-polarized macrophages toward dendritic cells.
- Author
-
Haegel H, Thioudellet C, Hallet R, Geist M, Menguy T, Le Pogam F, Marchand JB, Toh ML, Duong V, Calcei A, Settelen N, Preville X, Hennequi M, Grellier B, Ancian P, Rissanen J, Clayette P, Guillen C, Rooke R, and Bonnefoy JY
- Subjects
- Animals, Antibodies, Monoclonal immunology, Cell Differentiation immunology, Cell Line, Tumor, Cell Survival drug effects, Cell Survival immunology, Cells, Cultured, Chemokine CCL2 immunology, Chemokine CCL2 metabolism, Dendritic Cells immunology, Dendritic Cells metabolism, Flow Cytometry, HL-60 Cells, Humans, Interleukin-6 immunology, Interleukin-6 metabolism, Macrophage Colony-Stimulating Factor immunology, Macrophage Colony-Stimulating Factor metabolism, Macrophages immunology, Macrophages metabolism, Mice, Mice, Inbred BALB C, Monocytes immunology, Monocytes metabolism, NIH 3T3 Cells, Osteoclasts drug effects, Osteoclasts immunology, Osteoclasts metabolism, Osteolysis immunology, Phosphorylation drug effects, Protein Binding drug effects, Protein Binding immunology, Receptor, Macrophage Colony-Stimulating Factor immunology, Receptor, Macrophage Colony-Stimulating Factor metabolism, Signal Transduction drug effects, Signal Transduction immunology, Antibodies, Monoclonal pharmacology, Cell Differentiation drug effects, Dendritic Cells drug effects, Macrophages drug effects, Monocytes drug effects, Osteolysis prevention & control
- Abstract
Cancer progression has been associated with the presence of tumor-associated M2-macrophages (M2-TAMs) able to inhibit anti-tumor immune responses. It is also often associated with metastasis-induced bone destruction mediated by osteoclasts. Both cell types are controlled by the CD115 (CSF-1R)/colony-stimulating factor-1 (CSF-1, M-CSF) pathway, making CD115 a promising target for cancer therapy. Anti-human CD115 monoclonal antibodies (mAbs) that inhibit the receptor function have been generated in a number of laboratories. These mAbs compete with CSF-1 binding to CD115, dramatically affecting monocyte survival and preventing osteoclast and macrophage differentiation, but they also block CD115/CSF-1 internalization and degradation, which could lead to potent rebound CSF-1 effects in patients after mAb treatment has ended. We thus generated and selected a non-ligand competitive anti-CD115 mAb that exerts only partial inhibitory effects on CD115 signaling without blocking the internalization or the degradation of the CD115/CSF-1 complex. This mAb, H27K15, affects monocyte survival only minimally, but downregulates osteoclast differentiation and activity. Importantly, it inhibits monocyte differentiation to CD163(+)CD64(+) M2-polarized suppressor macrophages, skewing their differentiation toward CD14(-)CD1a(+) dendritic cells (DCs). In line with this observation, H27K15 also drastically inhibits monocyte chemotactic protein-1 secretion and reduces interleukin-6 production; these two molecules are known to be involved in M2-macrophage recruitment. Thus, the non-depleting mAb H27K15 is a promising anti-tumor candidate, able to inhibit osteoclast differentiation, likely decreasing metastasis-induced osteolysis, and able to prevent M2 polarization of TAMs while inducing DCs, hence contributing to the creation of more efficient anti-tumor immune responses.
- Published
- 2013
- Full Text
- View/download PDF
20. A heterologous prime/boost vaccination strategy enhances the immunogenicity of therapeutic vaccines for hepatitis C virus.
- Author
-
Fournillier A, Frelin L, Jacquier E, Ahlén G, Brass A, Gerossier E, Holmström F, Broderick KE, Sardesai NY, Bonnefoy JY, Inchauspé G, and Sällberg M
- Subjects
- Animals, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Genotype, Hepacivirus, Hepatitis C immunology, Immunization, Secondary, Interferon-gamma blood, Interleukin-2 blood, Mice, Mice, Inbred C57BL, Mice, Transgenic, Tumor Necrosis Factor-alpha blood, Vaccines, DNA immunology, Viral Hepatitis Vaccines genetics, Antibody Formation, Hepatitis C prevention & control, Vaccination methods, Viral Hepatitis Vaccines immunology
- Abstract
Background: We explored the concept of heterologous prime/boost vaccination using 2 therapeutic vaccines currently in clinical development aimed at treating chronically infected hepatitis C virus (HCV) patients: prime with a DNA-based vaccine expressing HCV genotype 1a NS3/4A proteins (ChronVac-C) and boost with a modified vaccinia virus Ankara vaccine expressing genotype 1b NS3/4/5B proteins (MVATG16643)., Methods: Two ChronVac-C immunizations 4 weeks apart were delivered intramuscularly in combination with in vivo electroporation and subsequently 5 or 12 weeks later boosted by 3 weekly subcutaneous injections of MVATG16643. Two mouse strains were used, and we evaluated quality, magnitude, and functionality of the T cells induced., Results: DNA prime/MVA boost regimen induced significantly higher levels of interferon γ (IFN-γ) or interleukin 2 (IL-2) ELISpot responses compared with each vaccine alone, independent of the time of analysis and the time interval between vaccinations. Both CD8⁺ and CD4⁺ T-cell responses as well as the spectrum of epitopes recognized was improved. A significant increase in polyfunctional IFN-γ/tumor necrosis factor α (TNF-α)/CD107a⁺ CD8⁺ T cells was detected following ChronVac-C/MVATG16643 vaccination (from 3% to 25%), and prime/boost was the only regimen that activated quadrifunctional T cells (IFN-γ/TNF-α/CD107a/IL-2). In vivo functional protective capacity of DNA prime/MVA boost was demonstrated in a Listeria-NS3-1a challenge model., Conclusions: We provide a proof-of-concept that immunogenicity of 2 HCV therapeutic vaccines can be improved using their combination, which merits further clinical development.
- Published
- 2013
- Full Text
- View/download PDF
21. Therapeutic cancer vaccines in the treatment of non-small-cell lung cancer.
- Author
-
Limacher JM, Spring-Giusti C, Bellon N, Ancian P, Rooke R, and Bonnefoy JY
- Subjects
- Cancer Vaccines administration & dosage, Cancer Vaccines immunology, Carcinoma, Non-Small-Cell Lung immunology, Carcinoma, Non-Small-Cell Lung pathology, Clinical Trials as Topic, Humans, Lung Neoplasms immunology, Lung Neoplasms pathology, Membrane Glycoproteins administration & dosage, Membrane Glycoproteins immunology, Membrane Glycoproteins therapeutic use, Cancer Vaccines therapeutic use, Carcinoma, Non-Small-Cell Lung therapy, Immunotherapy, Active methods, Lung Neoplasms therapy
- Abstract
Therapeutic vaccines are different from the well-known prophylactic vaccines in that they are designed to treat patients already suffering from a disease instead of preventing the disease in healthy individuals. Several therapeutic vaccines are today in late-stage clinical development for non-small-cell lung cancer. These vaccines use different approaches including peptides, cell lines and viral vectors, and explore different settings within the pathology. Some are given in monotherapy while others are combined with the classic therapies used with non-small-cell lung cancer. This review gives a summary of the therapeutic vaccines currently in late-stage clinical development for non-small-cell lung cancer.
- Published
- 2013
- Full Text
- View/download PDF
22. TLR2 ligation protects effector T cells from regulatory T-cell mediated suppression and repolarizes T helper responses following MVA-based cancer immunotherapy.
- Author
-
Amiset L, Fend L, Gatard-Scheikl T, Rittner K, Duong V, Rooke R, Muller S, Bonnefoy JY, Préville X, and Haegel H
- Abstract
Cancer immunotherapy is hampered by the immunosuppression maintained by regulatory T cells (Tregs) in tumor-bearing hosts. Stimulation of the Toll-like receptor 2 (TLR2) by Pam3Cys is known to affect Treg-mediated suppression. We found that Pam3Cys increases the proliferation of both CD4(+) effector T cells (Teffs) and Tregs co-cultured in vitro, but did not induce the proliferation of Tregs alone upon CD3 and CD28 stimulation. In a mouse model of RMA-MUC1 tumors, Pam3Cys was administered either alone or in combination with a modified vaccinia ankara (MVA)-based mucin 1 (MUC1) therapeutic vaccine. The combination of Pam3Cys with MVA-MUC1 (1) diminished splenic Treg/CD4(+) T-cell ratios to those found in tumor-free mice, (2) stimulated a specific anti-MUC1 interferon γ (IFNγ) response and (3) had a significant therapeutic effect on tumor growth and mouse survival. When CD4(+) Teffs and Tregs were isolated from Pam3Cys-treated mice, Teffs had become resistant to Treg-mediated suppression while upregulating the expression of BclL-x(L). Tregs from Pam3Cys-treated mice were fully suppressive for Teffs from naïve mice. Bcl-x(L) was induced by Pam3Cys with different kinetics in Tregs and Teffs. Teff from Pam3Cys-treated mice produced increased levels of Th1 and Th2-type cytokines and an interleukin (IL)-6-dependent secretion of IL-17 was observed in Teff:Treg co-cultures, suggesting that TLR2 stimulation had skewed the immune response toward a Th17 profile. Our results show for the first time that in a tumor-bearing host, TLR2 stimulation with Pam3Cys affects both Tregs and Teffs, protects Teff from Treg-mediated suppression and has strong therapeutic effects when combined with an MVA-based antitumor vaccine.
- Published
- 2012
- Full Text
- View/download PDF
23. [Cancer immunotherapy: recent breakthroughs and perspectives].
- Author
-
Tartour E, Sandoval F, Bonnefoy JY, and Fridman WH
- Subjects
- Angiogenesis Inhibitors therapeutic use, Animals, Antibodies, Monoclonal therapeutic use, Antigens, Neoplasm immunology, Antineoplastic Agents therapeutic use, Cancer Vaccines therapeutic use, Clinical Trials as Topic, Combined Modality Therapy, Forecasting, Humans, Immunity, Cellular, Immunotherapy, Active, Mice, Molecular Targeted Therapy, Neoplasm Proteins antagonists & inhibitors, Neoplasm Proteins immunology, Neoplasms drug therapy, Neoplasms radiotherapy, Precancerous Conditions therapy, Survival Analysis, T-Lymphocyte Subsets immunology, Therapies, Investigational, Immunotherapy trends, Neoplasms therapy
- Abstract
Immunotherapy of cancer has long been considered as an attractive therapeutic approach but with no impact on clinical practice. Two clinical protocols of immunotherapy, one based on a cancer vaccine in patients with prostate cancer or melanoma and the other using an immunomodulator targeting T cells (anti-CTLA4 mAb) in melanoma patients, have demonstrated clinical efficacy in two phase III clinical trials. To improve these encouraging clinical results, biomarkers to better select patients which may benefit from this therapy are actively searched. In addition, immunosuppression associated with cancer has to be overcome to allow a better immunostimulation. In contrast to chemotherapy, clinical variables to monitor the efficacy of immunotherapy has to be revisited and overall survival appears to be a better endpoint than clinical response defined by the RECIST criteria. Combination of immunotherapy with conventional treatments (chemotherapy, anti-angiogenic, etc.) should further improve this approach both in its effectiveness and in its clinical indications., (© 2011 médecine/sciences – Inserm / SRMS.)
- Published
- 2011
- Full Text
- View/download PDF
24. A poxvirus vaccine is safe, induces T-cell responses, and decreases viral load in patients with chronic hepatitis C.
- Author
-
Habersetzer F, Honnet G, Bain C, Maynard-Muet M, Leroy V, Zarski JP, Feray C, Baumert TF, Bronowicki JP, Doffoël M, Trépo C, Agathon D, Toh ML, Baudin M, Bonnefoy JY, Limacher JM, and Inchauspé G
- Subjects
- Adult, Antibodies, Viral blood, Dose-Response Relationship, Drug, Female, Genotype, Hepacivirus genetics, Hepatitis C, Chronic drug therapy, Humans, Interferon-gamma metabolism, Male, Middle Aged, RNA, Viral blood, T-Lymphocytes immunology, T-Lymphocytes pathology, Viral Nonstructural Proteins immunology, Viral Vaccines adverse effects, Viral Vaccines therapeutic use, Hepatitis C, Chronic pathology, Hepatitis C, Chronic virology, Poxviridae immunology, T-Lymphocytes drug effects, Viral Load drug effects, Viral Vaccines pharmacology
- Abstract
Background & Aims: Therapy for chronic hepatitis C (CHC) has limited efficacy, adverse effects, and high costs. Cohort and vaccine-based preclinical studies have indicated the importance of T-cell-based immunity in controlling viral infection. TG4040 is a recombinant poxvirus vaccine that expresses the hepatitis C virus (HCV) proteins NS3, NS4, and NS5B. We performed a phase I clinical trial to assess the safety, immunogenicity, and early antiviral efficacy of TG4040 in patients with CHC., Methods: In an open-label, dose-escalating study, patients with mild CHC (genotype 1) were assigned to 3 groups of 3 patients each; they received subcutaneous injections of 10⁶, 10⁷, or 10⁸ plaque-forming units of TG4040 on study days 1, 8, and 15. Six additional patients were given the highest dose of vaccine (10⁸ plaque-forming units). Patients were followed for 6 months after the last injection. T-cell-based and antibody responses and levels of HCV RNA were measured., Results: All 3 doses of TG4040 were well tolerated, without serious adverse events. Vaccine-induced HCV-specific cellular immune responses were observed in 5 of the 15 patients (33%). A transient decrease in circulating levels of HCV RNA, from -0.52 log₁₀ to -1.24 log₁₀, was observed in 8 patients; in 5 patients, the lowest level of HCV RNA was observed on day 37, after the first injection. The most pronounced decrease in viral load occurred in 2 patients, who also had marked vaccine-induced T-cell responses., Conclusions: In patients with CHC, the viral-vector-based vaccine TG4040 had a good safety profile, induced HCV-specific cellular immune responses, and reduced viral load. This vaccine should be investigated in further clinical studies, in combination with standard of care., (Copyright © 2011 AGA Institute. Published by Elsevier Inc. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
25. Vaccination against hepatitis B and C: towards therapeutic application.
- Author
-
Inchauspe G, Bach G, Martin P, and Bonnefoy JY
- Subjects
- CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes virology, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes virology, Hepatitis B epidemiology, Hepatitis B immunology, Hepatitis B Vaccines immunology, Hepatitis C epidemiology, Hepatitis C immunology, Humans, Vaccines, DNA immunology, Viral Hepatitis Vaccines immunology, Hepatitis B prevention & control, Hepatitis B Vaccines therapeutic use, Hepatitis C prevention & control, Vaccination, Vaccines, DNA therapeutic use, Viral Hepatitis Vaccines therapeutic use
- Published
- 2009
- Full Text
- View/download PDF
26. The biomarker revolution: a step toward personalized medicine.
- Author
-
Bonnefoy JY
- Published
- 2008
- Full Text
- View/download PDF
27. Clinical development of MVA-based therapeutic cancer vaccines.
- Author
-
Acres B and Bonnefoy JY
- Subjects
- Cancer Vaccines genetics, Humans, Neoplasms immunology, Antigens, Neoplasm genetics, Antigens, Neoplasm immunology, Cancer Vaccines immunology, Genetic Vectors, Neoplasms prevention & control, Neoplasms therapy, Vaccinia virus genetics
- Abstract
Tumor-associated antigens (TAAs) have been formulated into vaccines that, combined with adjuvants, cytokines or other strategies to boost the immune response, are now in clinical development. Both humoral and cellular immune responses to TAAs have been generated using these vaccines. This approach relies on the patient's own immune system generating an effective anti-tumor immune response. The advantage of this over therapy with monoclonal antibodies for the treatment of cancer is that multiple antigenic epitopes can be involved and the immune system is able to adapt to the most effective antigenic specificity for tumor growth control and rejection. In this article, we describe the clinical use of vaccinia virus, in particular modified vaccinia virus Ankara (MVA), to express TAAs in vivo and to stimulate an effective immune response to the cancer antigens.
- Published
- 2008
- Full Text
- View/download PDF
28. alphavbeta5 integrin sustains growth of human pre-B cells through an RGD-independent interaction with a basic domain of the CD23 protein.
- Author
-
Borland G, Edkins AL, Acharya M, Matheson J, White LJ, Allen JM, Bonnefoy JY, Ozanne BW, and Cushley W
- Subjects
- Amino Acid Motifs, Amino Acid Sequence, Binding Sites, Cells, Cultured, Humans, Immunoglobulin E metabolism, Molecular Sequence Data, Protein Structure, Tertiary, Receptors, Complement 3d metabolism, Receptors, IgE metabolism, B-Lymphocytes physiology, Hematopoietic Stem Cells physiology, Integrins physiology, Oligopeptides physiology, Receptors, IgE chemistry, Receptors, Vitronectin physiology
- Abstract
CD23 is a type II transmembrane glycoprotein synthesized by hematopoietic cells that has biological activity in both membrane-bound and freely soluble forms, acting via a number of receptors, including integrins. We demonstrate here that soluble CD23 (sCD23) sustains growth of human B cell precursors via an RGD-independent interaction with the alphavbeta5 integrin. The integrin recognizes a tripeptide motif in a small disulfide-bonded loop at the N terminus of the lectin head region of CD23, centered around Arg(172), Lys(173), and Cys(174) (RKC). This RKC motif is present in all forms of sCD23 with cytokine-like activity, and cytokine activity is independent of the lectin head, an "inverse RGD" motif, and the CD21 and IgE binding sites. RKC-containing peptides derived from this region of CD23 bind alphavbeta5 and are biologically active. The binding and activity of these peptides is unaffected by inclusion of a short peptide containing the classic RGD sequence recognized by integrins, and, in far-Western analyses, RKC-containing peptides bind to the beta subunit of the alphavbeta5 integrin. The interaction between alphavbeta5 and sCD23 indicates that integrins deliver to cells important signals initiated by soluble ligands without the requirement for interactions with RGD motifs in their common ligands. This mode of integrin signaling may not be restricted to alphavbeta5.
- Published
- 2007
- Full Text
- View/download PDF
29. Cancer vaccines: challenges and outlook in the field.
- Author
-
Paul S, Acres B, Limacher JM, and Bonnefoy JY
- Subjects
- Antigen Presentation drug effects, Antigens, Neoplasm drug effects, Drug Administration Routes, Drug Screening Assays, Antitumor, Humans, Neoplasms immunology, Neoplasms prevention & control, Vaccination methods, Vaccination trends, Adjuvants, Immunologic pharmacology, Cancer Vaccines therapeutic use, Drug Design, Neoplasms drug therapy
- Abstract
Therapeutic vaccination against cancer-associated antigens represents an attractive option for cancer therapy in view of its efficacy, the possibility of long-lasting immunity against the cancer, and safety profile. Nevertheless, it is now recognized that the same vaccination strategies used for prophylactic vaccinations against infectious diseases, cannot necessarily be used for therapeutic cancer vaccination. Cancer patients are usually immunosuppressed and most cancer-associated antigens are 'self-antigens', making it a significant challenge to vaccinate patients against a cancer-associated antigen. Various immunostimulation techniques are under investigation in an effort to bolster patients' immune systems and to overcome immune tolerance to self antigens. Strategies to stimulate antigen presentation, T-cell reactivity and innate immune activity are under investigation. Several clinical trials have been conducted to evaluate therapeutic cancer vaccines in patients, and attractive protocols, including those that combine the stimulation of specific T-cells and chemotherapy, or strategies to block immune regulation, are beginning to show some success. This feature review considers strategies for the development of effective therapeutic cancer vaccines, and highlights select vaccines that have already entered the clinic.
- Published
- 2007
30. Detection of epsilon class switching and IgE synthesis in human B cells.
- Author
-
Pène J, Guilhot F, Cognet I, Guglielmi P, Guay-Giroux A, Bonnefoy JY, Elson GC, Yssel H, and Gauchat JF
- Subjects
- B-Lymphocytes cytology, CD40 Antigens immunology, CD40 Ligand immunology, Cells, Cultured, Culture Techniques, DNA, Circular metabolism, Enzyme-Linked Immunosorbent Assay methods, Fetal Blood cytology, Humans, Immunoglobulin E genetics, Immunoglobulin epsilon-Chains genetics, Immunomagnetic Separation methods, Lymphocyte Activation, Palatine Tonsil cytology, Palatine Tonsil immunology, Spleen cytology, Spleen immunology, B-Lymphocytes immunology, Immunoglobulin Class Switching, Immunoglobulin E immunology, Immunoglobulin epsilon-Chains immunology
- Abstract
We observed that mast cells, as other cells expressing the CD40 ligand CD154, can trigger IgE synthesis in B cells in the presence of interleukin (IL)-4. Numerous complementary techniques can be used to follow the succession of molecular events leading to IgE synthesis. This chapter will illustrate how human B cells (naïve or memory) can be purified, stored, and cultivated in medium that is permissive for IgE synthesis and stimulated with IL-4 or IL-13 and CD40 activation, the latter being induced by soluble CD154, anti-CD40 antibodies, or CD154-expressing cells. All these molecules are expressed by mast cells. The quantification of the epsilon-sterile transcript synthesis by polymerase chain reaction or Northern blot, the epsilon excision circles produced during immunoglobulin heavy chain locus rearrangement by polymerase chain reaction, and the IgE production by enzyme-linked immunosorbent assay will be described.
- Published
- 2006
- Full Text
- View/download PDF
31. Cancer vaccines.
- Author
-
Bonnefoy JY
- Subjects
- Adenoviridae immunology, Adjuvants, Immunologic, Animals, Antigen Presentation, Antigens, Neoplasm administration & dosage, Antigens, Neoplasm immunology, Antigens, Neoplasm therapeutic use, Biomarkers, Dendritic Cells immunology, Dendritic Cells transplantation, Humans, Immune Tolerance, Immunization, Immunocompromised Host, Neoplasms immunology, Neoplasms prevention & control, Neoplasms, Experimental immunology, Neoplasms, Experimental therapy, Poxviridae immunology, T-Lymphocytes, Cytotoxic immunology, Tumor Escape, Vaccinia virus immunology, Cancer Vaccines administration & dosage, Cancer Vaccines immunology, Cancer Vaccines therapeutic use, Neoplasms therapy
- Abstract
Cancer vaccines - a dream or a reality? There is no doubt that a time will come when this new approach to cancer treatment will provide opportunities that will be both complementary and synergistic with existing treatments. Novelty is often linked to risk, and if there is a given medical field that desperately needs therapeutic improvement, cancer is it. Recent advances on the understanding of the functioning and manipulation of the immune system and on the host-tumour relationship, and on tumour escape, will soon pay off. However, the development of cancer vaccines is not an easy undertaking and the multitude of obstacles are presented and reviewed in this article. The cancer vaccine 'detractors' that are arguing negatively on the chances of success are, to this date, correct. Nevertheless, a parallel could be drawn from the monoclonal antibodies, which have suffered from a long and difficult history, to hold promise in man. Recent technological progress has once again demonstrated that one can eventually bypass the commonly accepted barriers, as there are now monoclonal-based products currently helping patients. The entire oncology community is eagerly awaiting the next chapter in the cancer vaccine story.
- Published
- 2004
- Full Text
- View/download PDF
32. Detection of circulating soluble CD28 in patients with systemic lupus erythematosus, primary Sjögren's syndrome and systemic sclerosis.
- Author
-
Hebbar M, Jeannin P, Magistrelli G, Hatron PY, Hachulla E, Devulder B, Bonnefoy JY, and Delneste Y
- Subjects
- Adolescent, Adult, CD28 Antigens genetics, Case-Control Studies, Cell Division drug effects, Cells, Cultured, Female, Humans, Interleukin-2 metabolism, Linear Models, Male, Middle Aged, Muromonab-CD3 pharmacology, RNA, Messenger analysis, Recombinant Proteins pharmacology, Statistics, Nonparametric, T-Lymphocytes cytology, T-Lymphocytes drug effects, T-Lymphocytes metabolism, CD28 Antigens blood, Lupus Erythematosus, Systemic immunology, Scleroderma, Systemic immunology, Sjogren's Syndrome immunology
- Abstract
The aim of this study was to evaluate the presence and the role of the serum soluble costimulatory molecule CD28 in patients with systemic lupus erythematosus (SLE), primary Sjögren's syndrome (SS), and systemic sclerosis (SSc). Soluble CD28 concentration was determined by ELISA in 45 patients with SLE, 45 patients with primary SS, 30 patients with SSc, and 45 healthy subjects. We also evaluated CD28 mRNA expression by semiquantitative RT-PCR, and the biological activity of recombinant soluble CD28 on T lymphocyte activity. Concentrations of soluble CD28 were significantly higher in patients with SLE, primary SS and SSc than in healthy subjects. Soluble CD28 concentrations were higher in patients with systemic primary SS than in patients with glandular-limited primary SS. PCR analysis suggested that soluble CD28 resulted from the shedding of the membrane form. In vitro assay revealed that soluble CD28 inhibits the anti-CD3 mAb induced T cell proliferation. Soluble CD28, which modulates the proliferation of T lymphocytes, could be associated with disease severity in patients with autoimmune disease, especially primary SS. These results suggest that soluble CD28 could play an important role in the regulation of autoimmune diseases.
- Published
- 2004
- Full Text
- View/download PDF
33. IL-18 is produced by prostate cancer cells and secreted in response to interferons.
- Author
-
Lebel-Binay S, Thiounn N, De Pinieux G, Vieillefond A, Debré B, Bonnefoy JY, Fridman WH, and Pagès F
- Subjects
- Caspase 1 genetics, Caspase 1 metabolism, Caspase Inhibitors, Cell Division drug effects, Cohort Studies, DNA Primers chemistry, Flow Cytometry, Gene Expression Regulation, Neoplastic, Humans, Immunoenzyme Techniques, Interleukin-18 Receptor alpha Subunit, Male, Prostate-Specific Antigen metabolism, Prostatic Neoplasms pathology, RNA, Messenger metabolism, Receptors, Interleukin metabolism, Receptors, Interleukin-18, Reverse Transcriptase Polymerase Chain Reaction, Tetrazolium Salts, Thiazoles, Tumor Cells, Cultured drug effects, Antineoplastic Agents pharmacology, Interferon-alpha pharmacology, Interferon-gamma pharmacology, Interleukin-18 biosynthesis, Prostatic Neoplasms metabolism
- Abstract
Murine models have shown that IL-18 has antiangiogenic and antitumor effects, but little is known about IL-18 production in human tumors. We investigated IL-18 expression in clinically localized prostate cancers by immunohistochemistry and showed that 75% of the prostate cancers studied (27/36 cases) presented with tumor cells producing IL-18. Prostate tumor cell lines PC-3, DU 145 and LNCaP synthesized the immature form of IL-18 (p24). IFN-gamma produced in prostate cancers induced caspase-1 mRNA and IL-18 secretion of tumor cell lines, which was inhibited by the cell-permeable Tyr-Val-Ala-Asp-aldehyde caspase-1 inhibitor (YVAD-CHO). Interestingly, IFN-alpha also induced IL-18 secretion of the poorly differentiated cell line PC-3. PC-3 and DU 145, but not the well-differentiated cell line LNCaP, expressed IL-18R alpha (IL-1Rrp) protein and transcripts for IL-18R beta (AcPL). Exogenous IL-18 increased mitochondrial activity of both cell lines evaluated by the tetrazolium (MTT) assay but did not influence their proliferation. This indicated that prostate tumor cells could secrete IL-18 in response to IFN-gamma in the tumor microenvironment and that IL-18 could act as a autocrine/paracrine factor for the tumor. In the cohort of patients studied, IL-18 expression in prostate cancers (with up to 10% of tumor cells stained) was associated with a favorable outcome and equally predictive as pathologic stage on multivariate analysis (log rank test, p = 0.02). Tumor IL-18 production is a novel physiopathologic feature of prostate cancer and appears to be a favorable event in the course of the disease. Modulation of IL-18 production by interferons could have a beneficial clinical effect, which deserves further investigation., (Copyright 2003 Wiley-Liss, Inc.)
- Published
- 2003
- Full Text
- View/download PDF
34. Enhanced pulmonary immunopathology following neonatal priming with formalin-inactivated respiratory syncytial virus but not with the BBG2NA vaccine candidate.
- Author
-
Plotnicky H, Siegrist CA, Aubry JP, Bonnefoy JY, Corvaïa N, Nguyen TN, and Power UF
- Subjects
- Animals, Animals, Newborn, Antibodies, Viral blood, CD4-Positive T-Lymphocytes immunology, Enzyme-Linked Immunosorbent Assay, Formaldehyde, Lung drug effects, Lymphocyte Activation, Mice, Mice, Inbred BALB C, Models, Animal, Respiratory Syncytial Virus Infections pathology, Respiratory Syncytial Virus Infections prevention & control, Vaccines, Inactivated toxicity, Lung pathology, Respiratory Syncytial Virus Infections immunology, Respiratory Syncytial Virus Vaccines immunology, Respiratory Syncytial Virus Vaccines toxicity, Vaccines, Inactivated immunology
- Abstract
Prevention of respiratory syncytial virus (RSV) disease will implicate neonatal priming. However, neonatal antigen exposure frequently results into Th2-like responses, some of which are critical for formalin-inactivated RSV (FI-RSV)-associated lung immunopathology. Neonatal immunization of mice may thus represent a more stringent model of RSV-enhanced pathology than adults. Indeed, after RSV challenge, lung cell infiltration, lymphocyte activation, and eosinophilia were higher following neonatal compared with adult FI-RSV priming of BALB/c mice. Unexpectedly, similar findings were obtained with Al(OH)(3)-adsorbed live RSV. In contrast, neonatal priming with BBG2Na, a recombinant RSV subunit vaccine candidate, formulated in either Al(OH)(3) or TiterMax (a Th1-driving adjuvant) resulted in predominant Th2- or Th1-like responses, respectively, but never elicited lung immunopathology post-challenge. Importantly, our data emphasize that the induction of Th2-like responses by RSV subunit vaccines do not necessarily imply lung immunopathology.
- Published
- 2003
- Full Text
- View/download PDF
35. The immunodominant influenza matrix T cell epitope recognized in human induces influenza protection in HLA-A2/K(b) transgenic mice.
- Author
-
Plotnicky H, Cyblat-Chanal D, Aubry JP, Derouet F, Klinguer-Hamour C, Beck A, Bonnefoy JY, and Corvaïa N
- Subjects
- Animals, Disease Models, Animal, Epitopes, T-Lymphocyte immunology, Female, HLA-A2 Antigen immunology, Humans, Immunologic Memory, Influenza, Human prevention & control, Lymphocyte Activation, Mice, Mice, Inbred C57BL, Mice, Transgenic, HLA-A2 Antigen genetics, Immunodominant Epitopes immunology, Influenza A virus immunology, Influenza Vaccines immunology, T-Lymphocytes, Cytotoxic immunology, Viral Matrix Proteins immunology
- Abstract
The protective efficacy of the influenza matrix protein epitope 58-66 (called M1), recognized in the context of human HLA-A2 molecules, was evaluated in a HLA-A2/K(b) transgenic mouse model of lethal influenza infection. Repeated subcutaneous immunizations with M1 increased the percentage of survival. This effect was mediated by T cells since protection was abolished following in vivo depletion of all T lymphocytes, CD8(+), or CD4(+) T cells. The survival correlated with the detection of memory CD8(+) splenocytes able to proliferate in vitro upon stimulation with M1 and to bind M1-loaded HLA-A2 dimers, as well as with M1-specific T cells in the lungs, which were directly cytotoxic to influenza-infected cells following influenza challenge. These results demonstrated for the first time that HLA-A2-restricted cytotoxic T cells specific for the major immunodominant influenza matrix epitope are protective against the infection. They encourage further in vivo evaluation of T cell epitopes recognized in the context of human MHC molecules.
- Published
- 2003
- Full Text
- View/download PDF
36. Outer membrane protein A (OmpA) activates human epidermal Langerhans cells.
- Author
-
Godefroy S, Corvaia N, Schmitt D, Aubry JP, Bonnefoy JY, Jeannin P, and Staquet MJ
- Subjects
- Antigens, CD biosynthesis, Antigens, CD1 biosynthesis, B7-2 Antigen, Bacterial Outer Membrane Proteins pharmacology, Cell Division immunology, Cell Movement drug effects, Cells, Cultured, Flow Cytometry, HLA-DR Antigens biosynthesis, Humans, Immunophenotyping, Langerhans Cells cytology, Langerhans Cells immunology, Membrane Glycoproteins biosynthesis, Protein Binding, T-Lymphocytes cytology, T-Lymphocytes immunology, Up-Regulation drug effects, Bacterial Outer Membrane Proteins metabolism, Langerhans Cells metabolism
- Abstract
Outer membrane protein (Omp)A is highly represented and conserved in the Enterobacteriaceae family. Using a recombinant OmpA from Klebsiella pneumoniae (kpOmpA), we have analysed the interaction between this bacterial cell wall protein and human Langerhans cells (LC), the antigen-presenting cells of the epidermis and mucosa. We showed that biotinylated kpOmpA binds to human LC freshly isolated from epidermis. kpOmpA up-regulated MHC class II, CD86 and CCR7 expression, enhanced migration in response to macrophage inflammatory protein-3beta (MIP-3beta) through a reconstituted basement membrane mimicking the prerequisite passage through the dermal-epidermal basement membrane on the way to lymph nodes. The allostimulatory function of kpOmpA-treated LC was more potent than that of untreated cells. Even though the proportion of LC which binds kpOmpA was shown to vary between individuals, our data indicate that kpOmpA binds to and activates LC, and suggest that recognition of OmpA by LC may be an initiating event in the antibacterial host response.
- Published
- 2003
- Full Text
- View/download PDF
37. Interferon-gamma switches monocyte differentiation from dendritic cells to macrophages.
- Author
-
Delneste Y, Charbonnier P, Herbault N, Magistrelli G, Caron G, Bonnefoy JY, and Jeannin P
- Subjects
- Animals, Antigens, CD analysis, Bone Marrow Cells cytology, Cell Culture Techniques methods, Cell Differentiation drug effects, Cytokines pharmacology, Gene Expression Regulation drug effects, Hematopoietic Stem Cells drug effects, Immunophenotyping, Interferon-gamma pharmacology, Interleukin-6 biosynthesis, Interleukin-6 pharmacology, Macrophage Colony-Stimulating Factor biosynthesis, Macrophage Colony-Stimulating Factor drug effects, Macrophage Colony-Stimulating Factor pharmacology, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Autocrine Communication, Dendritic Cells cytology, Interferon-gamma physiology, Macrophages cytology, Monocytes cytology
- Abstract
Human monocytes differentiate into dendritic cells (DCs) or macrophages according to the nature of environmental signals. Monocytes stimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF) plus interleukin 4 (IL-4) yield DCs. We tested here whether interferon-gamma (IFN-gamma), a potent activator of macrophages, may modulate monocyte differentiation. Addition of IFN-gamma to IL-4 plus GM-CSF-stimulated monocytes switches their differentiation from DCs to CD14(-)CD64(+) macrophages. IFN-gamma increases macrophage colony-stimulating factor (M-CSF) and IL-6 production by IL-4 plus GM-CSF-stimulated monocytes by acting at the transcriptional level and acts together with IL-4 to up-regulate M-CSF but not IL-6 production. IFN-gamma also increases M-CSF receptor internalization. Results from neutralizing experiments show that both M-CSF and IL-6 are involved in the ability of IFN-gamma to skew monocyte differentiation from DCs to macrophages. Finally, this effect of IFN-gamma is limited to early stages of differentiation. When added to immature DCs, IFN-gamma up-regulates IL-6 but not M-CSF production and does not convert them to macrophages, even in the presence of exogenous M-CSF. In conclusion, IFN-gamma shifts monocyte differentiation to macrophages rather than DCs through autocrine M-CSF and IL-6 production. These data show that IFN-gamma controls the differentiation of antigen-presenting cells and thereby reveals a new mechanism by which IFN-gamma orchestrates the outcome of specific immune responses.
- Published
- 2003
- Full Text
- View/download PDF
38. Outer membrane protein A (OmpA): a new pathogen-associated molecular pattern that interacts with antigen presenting cells-impact on vaccine strategies.
- Author
-
Jeannin P, Magistrelli G, Goetsch L, Haeuw JF, Thieblemont N, Bonnefoy JY, and Delneste Y
- Subjects
- Animals, Bacterial Outer Membrane Proteins metabolism, Bacterial Vaccines, Dendritic Cells immunology, Humans, Macrophages immunology, Membrane Glycoproteins metabolism, Receptors, Cell Surface metabolism, Signal Transduction, Toll-Like Receptor 2, Toll-Like Receptors, Antigen-Presenting Cells immunology, Bacterial Outer Membrane Proteins immunology
- Abstract
Outer membrane protein A (OmpA) is a class of proteins highly conserved among the Enterobacteriaceae family and throughout evolution. We have observed that antigen presenting cells (APCs) recognize and are activated by the recombinant OmpA from Klebsiella pneumoniae (KpOmpA). KpOmpA triggers cytokine production by macrophages and dendritic cells (DC), induces DC maturation and signals via Toll-like receptor 2. KpOmpA also interacts with endocytic receptor(s) expressed on DC and macrophages. Tumor antigens coupled to KpOmpA are taken up by APCs and gain access to the MHC class I pathway, triggering the initiation of protective anti-tumor cytotoxic responses in the absence of CD4 T cell help and adjuvant. Thus, OmpA appears as a new type of pathogen-associated molecular pattern (PAMP) usable as a vector in anti-infectious and therapeutic anti-tumor vaccines to elicit CTLs.
- Published
- 2002
- Full Text
- View/download PDF
39. Passive transfer of serum antibodies induced by BBG2Na, a subunit vaccine, in the elderly protects SCID mouse lungs against respiratory syncytial virus challenge.
- Author
-
Plotnicky-Gilquin H, Cyblat-Chanal D, Goetsch L, Lacheny C, Libon C, Champion T, Beck A, Pasche H, Nguyen TN, Bonnefoy JY, Bouveret-le-Cam N, and Corvaïa N
- Subjects
- Administration, Intranasal, Age Factors, Aged, Aged, 80 and over, Animals, Antibodies, Viral biosynthesis, Dose-Response Relationship, Immunologic, Female, HN Protein genetics, Humans, Immunization, Passive, Lung virology, Mice, Mice, Inbred BALB C, Mice, SCID, Middle Aged, Recombination, Genetic, Respiratory Syncytial Virus Infections virology, Respiratory Syncytial Virus Vaccines administration & dosage, Vaccines, Subunit immunology, Viral Envelope Proteins, Antibodies, Viral administration & dosage, Respiratory Syncytial Virus Infections prevention & control, Respiratory Syncytial Virus Vaccines immunology
- Abstract
Respiratory syncytial virus (RSV) is responsible for severe low respiratory tract infections in young infants and the elderly. To investigate whether BBG2Na, a recombinant subunit vaccine comprising aa 130-230 of the RSV G protein, induced protective Abs in subjects over 60 years during phase II clinical trial, pre- and postimmunization sera of individuals immunized with BBG2Na or placebo were transferred into SCID mice before RSV challenge. These sera dose-dependently reduced lung RSV titers. However at some points of serial dilutions, postimmunization sera of BBG2Na-immunized subjects only were significantly more efficient than the corresponding preimmunization sera, in agreement with the induction of an increased Ab response against multiple epitopes on RSV-A G protein. Thus, BBG2Na is immunogenic in the elderly and confers passive protection in mice after serum transfer. To our knowledge, this is the first description of protective Abs induced by a subunit vaccine in human.
- Published
- 2002
- Full Text
- View/download PDF
40. Gamma interferon-dependent protection of the mouse upper respiratory tract following parenteral immunization with a respiratory syncytial virus G protein fragment.
- Author
-
Plotnicky-Gilquin H, Cyblat-Chanal D, Aubry JP, Champion T, Beck A, Nguyen T, Bonnefoy JY, and Corvaïa N
- Subjects
- Adoptive Transfer, Amino Acid Sequence, Animals, CD4-Positive T-Lymphocytes cytology, CD4-Positive T-Lymphocytes immunology, Disease Models, Animal, Female, Humans, Mice, Mice, Inbred BALB C, Mice, Knockout, Mice, SCID, Molecular Sequence Data, Neutralization Tests, Peptide Fragments immunology, Respiratory System immunology, Th1 Cells immunology, Vaccination, Epitopes, T-Lymphocyte immunology, Immunity, Maternally-Acquired immunology, Interferon-gamma immunology, Lung immunology, Respiratory Syncytial Virus Infections prevention & control, Respiratory Syncytial Virus Vaccines immunology, Respiratory Syncytial Virus, Human immunology, Vaccines, Synthetic immunology, Viral Proteins immunology
- Abstract
The protective mechanisms induced in the mouse upper respiratory tract (URT) after intraperitoneal immunization with G2Na, a recombinant respiratory syncytial virus (RSV) G protein fragment (amino acid residues 130 to 230), were investigated. This protection was recently shown to be mediated by CD4(+) T cells and to be critically dependent on the cysteines and amino acids 193 and 194 (H. Plotnicky-Gilquin, A. Robert, L. Chevalet, J.-F. Haeuw, A. Beck, J.-Y. Bonnefoy, C. Brandt, C.-A. Siegrist, T. N. Nguyen, and U. F. Power, J. Virol. 74:3455-3463, 2000). On G2Na, we identified a domain (amino acid residues 182 to 198) responsible for the T-helper-cell activity. This region coincided with a peptide designed AICK (residues 184 to 198) which includes the previously identified murine and human T-helper-cell epitope on the native G protein (P. W. Tebbey, M. Hagen, and G. E. Hancock, J. Exp. Med. 188:1967-1972, 1998). Immunization with AICK, in alum or complete Freund's adjuvant, significantly reduced nasal RSV titers in normal BALB/c mice. However, although lung protection was induced, in contrast to the case with live RSV, neither AICK nor G2Na was able to prevent nasal infection in gamma interferon (IFN-gamma)-knockout mice. Anti-IFN-gamma neutralizing antibodies partially inhibited URT protection after administration to G2Na-immunized BALB/c mice. Furthermore, while purified CD4(+) T cells from BALB/c mice immunized with G2Na or AICK significantly reduced lung and nasal infection of naive recipient mice after adoptive transfer, the cells from IFN-gamma-knockout mice had no effect. Together, these results demonstrated for the first time that the T-helper-cell epitope of RSV G protein induces URT protection in mice after parenteral immunization through a Th1-type, IFN-gamma-dependent mechanism.
- Published
- 2002
- Full Text
- View/download PDF
41. DDA adjuvant induces a mixed Th1/Th2 immune response when associated with BBG2Na, a respiratory syncytial virus potential vaccine.
- Author
-
Klinguer-Hamour C, Libon C, Plotnicky-Gilquin H, Bussat MC, Revy L, Nguyen T, Bonnefoy JY, Corvaïa N, and Beck A
- Subjects
- Animals, Female, Interferon-gamma metabolism, Interleukin-4 metabolism, Interleukin-5 metabolism, Mice, Mice, Inbred BALB C, Quaternary Ammonium Compounds pharmacology, Rats, Respiratory Syncytial Virus Infections immunology, Respiratory Syncytial Virus Infections virology, Respiratory Syncytial Virus Vaccines adverse effects, Sigmodontinae, Vaccines, DNA adverse effects, Vaccines, DNA immunology, Viral Proteins genetics, Adjuvants, Immunologic, Respiratory Syncytial Virus Infections prevention & control, Respiratory Syncytial Virus Vaccines administration & dosage, Respiratory Syncytial Virus Vaccines immunology, Th1 Cells immunology, Th2 Cells immunology, Vaccines, DNA administration & dosage, Viral Proteins immunology
- Abstract
Human respiratory syncytial virus (hRSV) is one of the most common causes of respiratory infection in infants and the elderly. Previous attempts to vaccinate children against RSV failed and the induction of an aberrant Th2-type immune response was shown to induce severe to fatal pulmonary disease characterised in part by eosinophilia. BBG2Na is a promising human RSV subunit vaccine candidate which successfully passed phase II clinical trials in adults in association with Adju-Phos((R)). However, this formulation is not the most suitable for use in children since aluminium salts are known to induce a Th2-based immune response. In this study, we describe a potent and safe adjuvant formulation for BBG2Na in dimethyldioctadecylammonium bromide (DDA) that induces a mixed Th1/Th2 immune response in BALB/c mice. Furthermore, BBG2Na showed the same protective efficacy against RSV challenge when formulated either in DDA or in alum in mice and cotton rats.
- Published
- 2002
- Full Text
- View/download PDF
42. Measurement of nuclear factor-kappa B translocation on lipopolysaccharide-activated human dendritic cells by confocal microscopy and flow cytometry.
- Author
-
Blaecke A, Delneste Y, Herbault N, Jeannin P, Bonnefoy JY, Beck A, and Aubry JP
- Subjects
- Animals, Cell Nucleus drug effects, Cell Nucleus immunology, Cell Nucleus metabolism, Cell Separation, Cells, Cultured, Dendritic Cells drug effects, Dendritic Cells immunology, Dose-Response Relationship, Immunologic, Escherichia coli immunology, Humans, Lipopolysaccharides pharmacology, Lymphocyte Activation, Mice, Reproducibility of Results, Dendritic Cells metabolism, Flow Cytometry methods, Image Cytometry methods, Microscopy, Confocal, NF-kappa B metabolism
- Abstract
Background: Nuclear factor kappa B (NF-kappaB) is a ubiquitously expressed transcription factor that regulates cytokine and immunoglobulin (Ig) gene expression. In most cell types, the inactive p50/p65 NF-kappaB heterodimer is located in the cytoplasm, complexed to its IkappaB inhibitory unit. Stimulation of cells by various reagents such as bacterial endotoxin or cytokines leads to a dissociation of NF-kappaB from IkappaB and a rapid translocation of free NF-kappaB to the nucleus. The aim of this article is to define optimal conditions for the measurement of NF-kappaB translocation by both confocal microscopy and flow cytometry., Methods: Four commercial anti-NF-kappaB antibodies were evaluated by confocal microscopy, after using two methods of fixation and permeabilization of the cells. These antibodies were examined further by flow cytometry on purified nuclei., Results: Paraformaldehyde-methanol treatment of dendritic cells is a good combination to visualize NF-kappaB translocation by confocal microscopy. Three of the four antibodies tested gave good results on nonactivated and on lipopolysaccharide (LPS)-activated dendritic cells. The measurement of NF-kappaB translocation by flow cytometry on purified nuclei is a quick and sensitive method. Only one of the four evaluated antibodies showed a significant difference between nonactivated and activated cells. CONCLUSIONS; Microscopy and flow cytometry are quick and reproducible methods to measure NF-kappaB translocation and can be adapted to identify new molecules that activate dendritic cells., (Copyright 2002 Wiley-Liss, Inc.)
- Published
- 2002
- Full Text
- View/download PDF
43. Streptococcus pneumoniae polysaccharides conjugated to the outer membrane protein A from Klebsiella pneumoniae elicit protective antibodies.
- Author
-
Libon C, Haeuw JF, Crouzet F, Mugnier C, Bonnefoy JY, Beck A, and Corvaïa N
- Subjects
- Animals, Female, Immunoglobulin G biosynthesis, Immunoglobulin G classification, Immunoglobulin Isotypes biosynthesis, Immunoglobulin Isotypes classification, Klebsiella pneumoniae, Mice, Mice, Inbred BALB C, Streptococcus pneumoniae immunology, Bacterial Outer Membrane Proteins immunology, Pneumococcal Infections prevention & control, Pneumococcal Vaccines immunology, Polysaccharides immunology, Vaccines, Conjugate immunology
- Abstract
Polysaccharides (PSs) derived from Streptococcus pneumoniae include more than 90 serotypes and differ greatly in their immunogenicity. In addition, immunization with PSs does not induce high affinity antibody production and no memory B-cells are generated. Coupling PSs to carrier proteins has been reported to induce B-cell maturation and to install a B-cell memory. As an alternative carrier protein, the outer membrane protein A (OmpA) derived from Klebsiella pneumoniae has been coupled to various PSs. We evaluated the immunogenicity of two PS conjugates, using PS derived from S. pneumoniae types 14 and 19. In this report, we show that anti-PS IgG responses are generated after the conjugation of PSs to P40. In addition, the humoral response generated is able to protect mice from a bacterial challenge. Our results indicate that P40 could be included in the development of new PS conjugate vaccines.
- Published
- 2002
- Full Text
- View/download PDF
44. A role for CD147 in thymic development.
- Author
-
Renno T, Wilson A, Dunkel C, Coste I, Maisnier-Patin K, Benoit de Coignac A, Aubry JP, Lees RK, Bonnefoy JY, MacDonald HR, and Gauchat JF
- Subjects
- Amino Acid Sequence, Animals, Antibodies, Monoclonal metabolism, Base Sequence, Basigin, Binding Sites, Antibody genetics, Cell Cycle genetics, Cell Cycle immunology, Cell Differentiation genetics, Cell Differentiation immunology, Cell Line, Cloning, Molecular, Female, Fetus, Humans, Membrane Glycoproteins biosynthesis, Membrane Glycoproteins genetics, Membrane Glycoproteins immunology, Membrane Proteins genetics, Membrane Proteins immunology, Membrane Proteins metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Molecular Sequence Data, Organ Culture Techniques, Receptors, Antigen, T-Cell, alpha-beta biosynthesis, Thymus Gland cytology, Thymus Gland metabolism, Tumor Cells, Cultured, Antigens, CD, Antigens, Neoplasm, Antigens, Surface, Avian Proteins, Blood Proteins, Membrane Glycoproteins physiology, Thymus Gland embryology, Thymus Gland immunology
- Abstract
We have previously identified a mAb that binds to a molecule expressed preferentially on the surface of cycling thymocytes. In this study the molecule recognized by this mAb has been identified in the mouse as CD147 (basigin) by expression cloning. We show that CD147 expression correlates with cycling of immature thymocytes even in the absence of TCRbeta selection and that ligation of this molecule on immature fetal thymocytes inhibits their further development into mature T cells.
- Published
- 2002
- Full Text
- View/download PDF
45. Functionally active fusion protein of the novel composite cytokine CLC/soluble CNTF receptor.
- Author
-
Guillet C, Lelièvre E, Plun-Favreau H, Froger J, Chabbert M, Hermann J, Benoit de Coignac A, Bonnefoy JY, Gascan H, Gauchat JF, and Elson G
- Subjects
- Amino Acid Sequence, Base Sequence, Cell Line, Cell Separation, Chromatography, Gel, DNA Primers, Flow Cytometry, Humans, Molecular Sequence Data, Phosphorylation, Recombinant Fusion Proteins metabolism, Signal Transduction, Cytokines metabolism, Receptor, Ciliary Neurotrophic Factor metabolism
- Abstract
The heterodimeric cytokine composed of the soluble ciliary neurotrophic factor receptor (sCNTFR) and the IL-6 family member cardiotrophin-like cytokine (CLC) was recently identified as a new ligand for gp130-leukemia inhibitory factor receptor (LIFR) complex [Plun-Favreau, H., Elson, G., Chabbert, M., Froger, J., deLapeyriere, O., Lelievre, E., Guillet, C., Hermann, J., Gauchat, J. F., Gascan, H. & Chevalier, S. (2001) EMBO J. 20, 1692-1703]. This heterodimer shows overlapping biological properties with LIF. Although CLC contains a putative signal peptide and therefore should enter into the classical secretory pathway, the protein has been shown to be retained within transfected mammalian cells, unless coexpressed with either sCNTFR or cytokine like factor (CLF) [Elson, G. C., Lelievre, E., Guillet, C., Chevalier, S., Plun-Favreau, H., Froger, J., Suard, I., de Coignac, A. B., Delneste, Y., Bonnefoy, J. Y., Gauchat, J. F. & Gascan, H. (2000) Nat. Neurosci. 3, 867-872]. In the present study, we demonstrate that a fusion protein comprising CLC covalently coupled through a glycine/serine linker to sCNTFR (CC-FP) is efficiently secreted from transfected mammalian cells. CC-FP shows enhanced activities in respect to the CLC/sCNTFR native complex, on a number of cells expressing gp130 and LIFR on their surface. In addition, CC-FP is able to compete with CNTF for cell binding, indicating that both cytokines share binding epitope(s) expressed by their receptor complex. Analysis of the downstream signaling events revealed the recruitment by CC-FP of the signal transducer and activator of transcription (STAT)-3, Akt and mitogen-activated protein (MAP) kinase pathways. The monomeric bioactive CLC/sCNTFR fusion protein is therefore a powerful tool to study the biological role of the recently described cytokine CLC.
- Published
- 2002
- Full Text
- View/download PDF
46. Safety and immunogenicity of a novel recombinant subunit respiratory syncytial virus vaccine (BBG2Na) in healthy young adults.
- Author
-
Power UF, Nguyen TN, Rietveld E, de Swart RL, Groen J, Osterhaus AD, de Groot R, Corvaia N, Beck A, Bouveret-Le-Cam N, and Bonnefoy JY
- Subjects
- Adolescent, Adult, Antibodies, Viral biosynthesis, Antigens, Viral adverse effects, Antigens, Viral immunology, Epitopes immunology, Humans, Middle Aged, Peptides immunology, Recombinant Proteins adverse effects, Recombinant Proteins immunology, Respiratory Syncytial Virus Infections etiology, Respiratory Tract Infections etiology, Vaccines, Subunit adverse effects, Vaccines, Subunit immunology, Viral Proteins adverse effects, Viral Proteins immunology, Respiratory Syncytial Virus Vaccines adverse effects, Respiratory Syncytial Virus Vaccines immunology
- Abstract
A novel recombinant respiratory syncytial virus (RSV) subunit vaccine, designated BBG2Na, was administered to 108 healthy adults randomly assigned to receive 10, 100, or 300 microg of BBG2Na in aluminum phosphate or saline placebo. Each subject received 1, 2, or 3 intramuscular injections of the assigned dose at monthly intervals. Local and systemic reactions were mild, and no evidence of harmful properties of BBG2Na was reported. The highest ELISA and virus-neutralizing (VN) antibody responses were evident in the 100- and 300-microg groups; second or third injections provided no significant boosts against RSV-derived antigens. BBG2Na induced > or 2-fold and > or =4-fold increases in G2Na-specific ELISA units in up to 100% and 57% of subjects, respectively; corresponding RSV-A-specific responses were 89% and 67%. Furthermore, up to 71% of subjects had > or =2-fold VN titer increases. Antibody responses to 2 murine lung protective epitopes were also highly boosted after vaccination. Therefore, BBG2Na is safe, well tolerated, and highly immunogenic in RSV-seropositive adults.
- Published
- 2001
- Full Text
- View/download PDF
47. Differential histopathology and chemokine gene expression in lung tissues following respiratory syncytial virus (RSV) challenge of formalin-inactivated RSV- or BBG2Na-immunized mice.
- Author
-
Power UF, Huss T, Michaud V, Plotnicky-Gilquin H, Bonnefoy JY, and Nguyen TN
- Subjects
- Animals, Chemokine CCL2 genetics, Chemokine CCL4, Chemokine CCL5 genetics, Chemokine CXCL2, Female, Immunization, Lung metabolism, Macrophage Inflammatory Proteins genetics, Mice, Mice, Inbred BALB C, Reverse Transcriptase Polymerase Chain Reaction, Vaccines, Inactivated immunology, Vaccines, Subunit immunology, Chemokines genetics, Lung pathology, Respiratory Syncytial Virus Vaccines immunology
- Abstract
A BALB/c mouse model of enhanced pulmonary pathology following vaccination with formalin-inactivated alum-adsorbed respiratory syncytial virus (FI-RSV) and live RSV challenge was used to determine the type and kinetics of histopathologic lesions induced and chemokine gene expression profiles in lung tissues. These data were compared and contrasted with data generated following primary and/or secondary RSV infection or RSV challenge following vaccination with a promising subunit vaccine, BBG2Na. Severe peribronchiolitis and perivascularitis coupled with alveolitis and interstitial inflammation were the hallmarks of lesions in the lungs of FI-RSV-primed mice, with peak histopathology evident on days 5 and 9. In contrast, primary RSV infection resulted in no discernible lesions, while challenge of RSV-primed mice resulted in rare but mild peribronchiolitis and perivascularitis, with no evidence of alveolitis or interstitial inflammation. Importantly, mice vaccinated with a broad dose range (20 to 0.02 microg) of a clinical formulation of BBG2Na in aluminium phosphate demonstrated histopathology similar to that observed in secondary RSV infection. At the molecular level, FI-RSV priming was characterized by a rapid and strong up-regulation of eotaxin and monocyte chemotactic protein 3 (MCP-3) relative gene expression (potent lymphocyte and eosinophil chemoattractants) that was sustained through late time points, early but intermittent up-regulation of GRO/melanoma growth stimulatory activity gene and inducible protein 10 gene expression, while macrophage inflammatory protein 2 (MIP-2) and especially MCP-1 were up-regulated only at late time points. By comparison, primary RSV infection or BBG2Na priming resulted in considerably lower eotaxin and MCP-3 gene expression increases postchallenge, while expression of lymphocyte or monocyte chemoattractant chemokine genes (MIP-1beta, MCP-1, and MIP-2) were of higher magnitude and kinetics at early, but not late, time points. Our combined histopathologic and chemokine gene expression data provide a basis for differentiating between aberrant FI-RSV-induced immune responses and normal responses associated with RSV infection in the mouse model. Consequently, our data suggest that BBG2Na may constitute a safe RSV subunit vaccine for use in seronegative infants.
- Published
- 2001
- Full Text
- View/download PDF
48. Targeting of nasal mucosa-associated antigen-presenting cells in vivo with an outer membrane protein A derived from Klebsiella pneumoniae.
- Author
-
Goetsch L, Gonzalez A, Plotnicky-Gilquin H, Haeuw JF, Aubry JP, Beck A, Bonnefoy JY, and Corvaïa N
- Subjects
- Administration, Intranasal, Amino Acid Sequence, Animals, Antibodies, Bacterial blood, Antibodies, Bacterial immunology, Antibodies, Viral blood, Antibodies, Viral immunology, Bacterial Outer Membrane Proteins genetics, Bacterial Outer Membrane Proteins metabolism, Biological Transport, Disease Models, Animal, Female, Humans, Immunity, Mucosal, Lymphoid Tissue immunology, Mice, Mice, Inbred BALB C, Molecular Sequence Data, Nasal Mucosa immunology, Recombinant Fusion Proteins immunology, Recombinant Fusion Proteins metabolism, Respiratory Syncytial Virus Infections immunology, Vaccination methods, Adjuvants, Immunologic, Antigen-Presenting Cells immunology, Antigens, Viral immunology, Bacterial Outer Membrane Proteins immunology, Klebsiella pneumoniae immunology, Respiratory Syncytial Virus Infections prevention & control, Respiratory Syncytial Virus Vaccines immunology, Vaccines, Synthetic immunology, Viral Proteins immunology
- Abstract
Administration of vaccines by the nasal route has recently proven to be one of the most efficient ways for inducing both mucosal and systemic antibody responses in experimental animals. Our results demonstrate that P40, a well-defined outer membrane protein A from Klebsiella pneumoniae, is indeed a carrier molecule suitable for nasal immunization. Using fragments from the respiratory syncytial virus subgroup A (RSV-A) G protein as antigen models, it has been shown that P40 is able to induce both systemic and mucosal immunity when fused or coupled to a protein or a peptide and administered intranasally (i.n.) to naive or K. pneumoniae-primed mice. Confocal analyses of nasal mucosa-associated lymphoid tissue after i.n. instillation of P40 showed that this molecule is able to cross the nasal epithelium and target CD11c-positive cells likely to be murine dendritic cells or macrophages. More importantly, this targeting of antigen-presenting cells following i.n. immunization with a subunit of the RSV-A molecule in the absence of any mucosal adjuvant results in both upper and lower respiratory tract protection against RSV-A infection.
- Published
- 2001
- Full Text
- View/download PDF
49. Histamine polarizes human dendritic cells into Th2 cell-promoting effector dendritic cells.
- Author
-
Caron G, Delneste Y, Roelandts E, Duez C, Bonnefoy JY, Pestel J, and Jeannin P
- Subjects
- Cell Differentiation, Cells, Cultured, Dendritic Cells drug effects, Humans, Hypersensitivity immunology, Interferon-gamma biosynthesis, Interleukin-12 biosynthesis, Lipopolysaccharides pharmacology, Myeloid Progenitor Cells drug effects, Myeloid Progenitor Cells immunology, Dendritic Cells immunology, Histamine pharmacology, Th2 Cells immunology
- Abstract
Allergic disorders are characterized by allergen-specific Th2-biased responses. Signals controlling Th2 cell polarization, especially those acting by polarizing dendritic cells (DC) into Th2-promoting DC (DC2), are not well known. Histamine, a mediator released by allergen-stimulated mast cells from allergic subjects, has been reported to activate human immature DC. We have therefore tested whether histamine affects DC polarization. We report here that histamine inhibits LPS-induced IL-12 production and polarizes uncommitted maturing DC into effector DC2. DC matured in the presence of histamine fail to produce IL-12 upon subsequent stimulation and prime Th2 responses, even in presence of IFN-gamma, a potent DC1-driving factor. All these effects are mediated through both H1 and H2 receptors. These data show that histamine is a potent DC2-polarizing factor and provide evidence for a novel mechanism that explains the initiation and maintenance of a predominant Th2 response in allergic disorders.
- Published
- 2001
- Full Text
- View/download PDF
50. Stability and CTL-activity of P40/ELA melanoma vaccine candidate.
- Author
-
Beck A, Goetsch L, Champion T, Bussat MC, Aubry JP, Klinguer-Hamour C, Haeuw JF, Bonnefoy JY, and Corvaïa N
- Subjects
- Animals, Cancer Vaccines chemistry, Cancer Vaccines therapeutic use, Chromatography, High Pressure Liquid, Melanoma immunology, Mice, Mice, Transgenic, Models, Molecular, Cancer Vaccines immunology, Melanoma therapy, T-Lymphocytes, Cytotoxic immunology
- Abstract
The decapeptide ELA (ELAGIGILTV), a Melan-A/MART-1 antigen immunodominant peptide analogue, is an interesting melanoma vaccine candidate alone or in combination with other tumour antigens. P40, the recombinant outer membrane protein A of Klebsiella pneumoniae (kpOmpA), was recently shown to target dendritic cells and to induce peptide-specific CTLs. Here we investigated the adjuvant role of P40 mixed or chemically conjugated to ELA. This compound is an N-terminal glutamic acid-containing peptide. However, it has been reported that the amino group and the gamma-carboxylic group of glutamic acids easily condense to form pyroglutamic derivatives. Usually, to overcome this stability problem, peptides of pharmaceutical interest were developed with a pyroglutamic acid instead of N-terminal glutamic acid, without loss of pharmacological properties. Unfortunately, the pyroglutamic acid derivative (PyrELA) as well as the N-terminal acetyl capped derivative (AcELA) failed to elicit CTL activity when mixed with P40 adjuvant protein. Despite the apparent minor modifications introduced by PyrELA and AcELA, these two derivatives have probably lower affinity than ELA for the class I Major Histocompatibility Complex. Furthermore, this stability problem is worse in the case of clinical grade ELA, produced as an acetate salt, like most of the pharmaceutical grade peptides. We report here that the hydrochloride shows a higher stability than the acetate and may be suitable for use in man., (Copyright 2001 The International Association for Biologicals.)
- Published
- 2001
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.