In the past decade, single-cell transcriptomics has helped to uncover new cell types and states and led to the construction of a cellular compendium of health and disease. Despite this progress, some difficult-to-sequence cells remain absent from tissue atlases. Eosinophils-elusive granulocytes that are implicated in a plethora of human pathologies1-5-are among these uncharted cell types. The heterogeneity of eosinophils and the gene programs that underpin their pleiotropic functions remain poorly understood. Here we provide a comprehensive single-cell transcriptomic profiling of mouse eosinophils. We identify an active and a basal population of intestinal eosinophils, which differ in their transcriptome, surface proteome and spatial localization. By means of a genome-wide CRISPR inhibition screen and functional assays, we reveal a mechanism by which interleukin-33 (IL-33) and interferon-γ (IFNγ) induce the accumulation of active eosinophils in the inflamed colon. Active eosinophils are endowed with bactericidal and T cell regulatory activity, and express the co-stimulatory molecules CD80 and PD-L1. Notably, active eosinophils are enriched in the lamina propria of a small cohort of patients with inflammatory bowel disease, and are closely associated with CD4+ T cells. Our findings provide insights into the biology of eosinophils and highlight the crucial contribution of this cell type to intestinal homeostasis, immune regulation and host defence. Furthermore, we lay a framework for the characterization of eosinophils in human gastrointestinal diseases.
- MeSH
- Single-Cell Gene Expression Analysis MeSH
- B7-1 Antigen metabolism MeSH
- Eosinophils * classification cytology immunology metabolism MeSH
- Inflammatory Bowel Diseases immunology MeSH
- Immunity * MeSH
- Interferon-gamma MeSH
- Interleukin-33 MeSH
- Colitis * immunology pathology MeSH
- Humans MeSH
- Mice MeSH
- Proteome MeSH
- Intestines * immunology pathology MeSH
- T-Lymphocytes MeSH
- Transcriptome MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
OBJECTIVE: We measured and compared changes in the percentage of cells expressing CD80, CD86, CD40, HLA-DR and the expression of these molecules on B cells and monocytes of patients who underwent either on-pump, mini on-pump or off-pump cardiac surgery. METHODS: Blood samples from patients who underwent either on-pump, mini on-pump or off-pump cardiac surgery were collected before surgery, instantly after surgery and on the 1(st), 3(rd) and 7(th) days after surgery. Surface expression of CD80, CD86, CD40 and HLA-DR molecules was determined by flow cytometry. RESULTS: Our results show that all three surgical techniques altered the expression of these molecules, as well as the percentage relative number of specific cell populations. We identified statistically significant differences when comparing different surgical techniques. On-pump surgery revealed a more pronounced impact on the phenotype of immune system cells than the other techniques. Therefore, it is likely that the function of immune cells is changed the most by on-pump surgery. We found a lower decrease in the number of CD80(+) monocytes and a lower drop in the CD40 expression on monocytes in off-pump patients in comparison with on-pump patients. CONCLUSION: All the types of cardiac surgical techniques, off-pump, on-pump and modified mini-invasive on-pump, are associated with changes in CD80, CD86, CD40 and HLA-DR expression. We found several significant differences in the expression of the selected molecules when we compared all three groups of patients.
- MeSH
- CD40 Antigens analysis MeSH
- B7-1 Antigen analysis MeSH
- B7-2 Antigen analysis MeSH
- B-Lymphocytes immunology MeSH
- HLA-DR Antigens analysis MeSH
- Cardiac Surgical Procedures * MeSH
- Middle Aged MeSH
- Humans MeSH
- Monocytes immunology MeSH
- Prospective Studies MeSH
- Aged MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Keywords
- Coleyovy toxiny, chimérický antigenní receptor,
- MeSH
- CTLA-4 Antigen antagonists & inhibitors administration & dosage therapeutic use MeSH
- Antigens, CD19 MeSH
- Antigens, CD20 MeSH
- Programmed Cell Death 1 Receptor antagonists & inhibitors administration & dosage therapeutic use MeSH
- CD28 Antigens MeSH
- B7-1 Antigen antagonists & inhibitors therapeutic use MeSH
- Autoimmune Diseases complications prevention & control MeSH
- T-Lymphocytes, Cytotoxic immunology MeSH
- Dendritic Cells immunology MeSH
- Hematologic Neoplasms diagnosis therapy MeSH
- Immune System Phenomena drug effects MeSH
- Immunologic Techniques MeSH
- Immunotherapy, Adoptive * history methods utilization MeSH
- Immunotherapy * history methods utilization MeSH
- Remission Induction MeSH
- Medical Oncology * MeSH
- Humans MeSH
- Melanoma immunology therapy MeSH
- Antibodies, Monoclonal pharmacology adverse effects therapeutic use MeSH
- Prostatic Neoplasms diagnosis immunology therapy MeSH
- Neoplasms * history immunology therapy MeSH
- Cancer Vaccines * history immunology therapeutic use MeSH
- In Vitro Techniques utilization MeSH
- Lymphocytes, Tumor-Infiltrating immunology MeSH
- Check Tag
- Humans MeSH
Deciphering the mechanisms that allow the induction of strong immune responses is crucial to developing efficient vaccines against infectious diseases and cancer. Based on the discovery that the adenylate cyclase from Bordetella pertussis binds to the CD11b/CD18 integrin, we developed a highly efficient detoxified adenylate cyclase-based vector (CyaA) capable of delivering a large variety of Ags to the APC. This vector allows the induction of protective and therapeutic immunity against viral and tumoral challenges as well as against transplanted tumors in the absence of any added adjuvant. Two therapeutic vaccine candidates against human papilloma viruses and melanoma have been developed recently, based on the CyaA vector, and are currently in clinical trials. We took advantage of one of these highly purified vaccines, produced under good manufacturing practice-like conditions, to decipher the mechanisms by which CyaA induces immune responses. In this study, we demonstrate that CyaA binds both human and mouse CD11b(+) dendritic cells (DCs) and induces their maturation, as shown by the upregulation of costimulatory and MHC molecules and the production of proinflammatory cytokines. Importantly, we show that DCs sense CyaA through the TLR4/Toll/IL-1R domain-containing adapter-inducing IFN-β pathway, independent of the presence of LPS. These findings show that CyaA possesses the intrinsic ability to not only target DCs but also to activate them, leading to the induction of strong immune responses. Overall, this study demonstrates that Ag delivery to CD11b(+) DCs in association with TLR4/Toll/IL-1R domain-containing adapter-inducing IFN-β activation is an efficient strategy to promote strong specific CD8(+) T cell responses.
- MeSH
- Adaptor Proteins, Vesicular Transport immunology MeSH
- Adenylate Cyclase Toxin immunology MeSH
- CD11b Antigen immunology MeSH
- B7-1 Antigen biosynthesis MeSH
- B7-2 Antigen biosynthesis MeSH
- Bordetella pertussis immunology MeSH
- Cell Differentiation immunology MeSH
- T-Lymphocytes, Cytotoxic immunology MeSH
- Dendritic Cells cytology immunology MeSH
- Interferon-beta immunology MeSH
- Interleukin-1beta biosynthesis MeSH
- Interleukin-6 biosynthesis MeSH
- Cells, Cultured MeSH
- Mice, Inbred C57BL MeSH
- Mice, Knockout MeSH
- Mice MeSH
- Receptor, Interferon alpha-beta genetics MeSH
- Receptors, Interleukin-1 immunology MeSH
- Signal Transduction immunology MeSH
- Tumor Necrosis Factor-alpha biosynthesis MeSH
- Toll-Like Receptor 4 immunology MeSH
- Tyrosine genetics MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Revmatoidní artritida (RA) je chronické zánětlivé onemocnění provázené bolestí, únavou a progredujícím omezením fyzické funkce, jež vedou ke zhoršení kvality života pacientů. V současné době je k terapii RA dostupných osm biologických přípravků – monoklonální protilátky namířené vůči TNF-α adalimumab, etanercept, infliximab, certolizumab a golimumab, rekombinantní fúzní protein anti-CD80/86 abatacept, monoklonální protilátka anti-CD20 rituximab a monoklonální protilátka proti receptorům pro IL-6 tocilizumab. Dosud však nebyla provedena žádná studie, která by přímo (tzv. head-to-head) porovnala účinnost různých biologických léků v kombinaci s methotrexátem (MTX) u nemocných s RA, u nichž selhala léčba MTX a kteří dosud nebyli léčeni biologickou terapií.
- MeSH
- B7-1 Antigen MeSH
- Biological Therapy MeSH
- Drug Evaluation MeSH
- Immunoconjugates * administration & dosage pharmacokinetics adverse effects therapeutic use MeSH
- Immunosuppressive Agents MeSH
- Clinical Trials, Phase III as Topic MeSH
- Drug Therapy, Combination MeSH
- Humans MeSH
- Methotrexate administration & dosage pharmacokinetics adverse effects therapeutic use MeSH
- Randomized Controlled Trials as Topic MeSH
- Medication Reconciliation MeSH
- Arthritis, Rheumatoid * drug therapy MeSH
- Tumor Necrosis Factor-alpha * antagonists & inhibitors administration & dosage pharmacokinetics adverse effects therapeutic use MeSH
- Check Tag
- Humans MeSH
Treatment of murine EL4 T cell lymphoma with N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer conjugates of doxorubicin (Dox) leads to complete tumor regression and to the development of therapy-dependent longlasting cancer resistance. This phenomenon occurs with two types of Dox conjugates tested, despite differences in the covalent linkage of Dox to the polymer carrier. Such a cancer resistance cannot fully express in conventional treatment with free Dox, due to substantial immunotoxicity of the treatment, which was not observed in the polymer conjugates. In this study, calreticulin (CRT) translocation and high mobility group box-1 protein (HMGB1) release was observed in EL4 cells treated with a conjugate releasing Dox by a pH-dependent manner. As a result, the treated tumor cells were engulfed by dendritic cells (DC) in vitro, and induced their expression of CD80, CD86, and MHC II maturation markers. Conjugates with Dox bound via an amide bond only increased translocation of HSPs to the membrane, which led to an elevated phagocytosis but was not sufficient to induce increase of the maturation markers on DCs in vitro. Both types of conjugates induced engulfment of the target tumor cells in vivo, that was more intense than that seen with free Dox. It means that the induction of anti-tumor immunity documented upon treatment of EL4 lymphoma with HPMA-bound Dox conjugates does not rely solely on CRT-mediated cell death, but involves multiple mechanisms.
- MeSH
- B7-1 Antigen metabolism MeSH
- B7-2 Antigen metabolism MeSH
- Apoptosis drug effects MeSH
- Drug Resistance, Neoplasm drug effects MeSH
- Dendritic Cells cytology immunology MeSH
- Doxorubicin administration & dosage analogs & derivatives chemistry toxicity MeSH
- Phagocytosis MeSH
- Calreticulin metabolism MeSH
- Hydrogen-Ion Concentration MeSH
- Polymethacrylic Acids administration & dosage chemistry toxicity MeSH
- Lymphoma, T-Cell drug therapy immunology MeSH
- Mice, Inbred C57BL MeSH
- Mice MeSH
- Cell Line, Tumor MeSH
- Drug Carriers chemistry MeSH
- HMGB1 Protein metabolism MeSH
- Heat-Shock Proteins metabolism MeSH
- Antineoplastic Agents administration & dosage chemistry toxicity MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
N-acetyl-D-glucosamine-coated polyamidoamine dendrimer (GN8P), exerting high binding affinity to rodent recombinant NKR-P1A and NKR-P1C activating proteins, was shown previously to delay the development of rat colorectal carcinoma as well as mouse B16F10 melanoma, and to potentiate antigen-specific antibody formation in healthy C57BL/6 mice via NK cell stimulation. In this study, we investigated whether GN8P also modulates tumor-specific B cell responses. Serum anti-B16F10 melanoma IgG levels, IgG2a mRNA expression, antibody dependent cell-mediated cytotoxicity (ADCC), and counts of plasma as well as antigen presenting B cells were evaluated in tumor-bearing C57BL/6 mice treated with GN8P and in respective controls. To reveal the mechanism of GN8P effects, the synthesis of interferon-gamma (IFN-γ) and interleukin-4 (IL-4), cytokines involved in regulation of immunoglobulin class switch, was determined. The GN8P treatment significantly elevated IgG, and particularly IgG2a, response against B16F10 melanoma, which led to augmented ADCC reaction. The significant increase in production of IFN-γ, which is known to support IgG2a secretion, was observed solely in NK1.1 expressing cell populations, predominantly in NK cells. Moreover, GN8P raised the number of plasma cells, and promoted antigen presenting capacity of I-A/I-E-positive B lymphocytes by up-regulation of their CD80 and CD86 co-stimulatory molecule expression. These results indicate that GN8P-induced enhancement of tumor-specific antibody formation is triggered by NK cell activation, and contributes to complexity of anticancer immune response involving lectin-saccharide interaction.
- MeSH
- Acetylglucosamine chemistry pharmacology MeSH
- B7-1 Antigen biosynthesis genetics MeSH
- B7-2 Antigen biosynthesis genetics MeSH
- B-Lymphocytes drug effects immunology metabolism MeSH
- Antibody-Dependent Cell Cytotoxicity drug effects immunology MeSH
- Killer Cells, Natural drug effects immunology metabolism MeSH
- Dendrimers chemistry pharmacology MeSH
- Immunoglobulin G biosynthesis blood genetics immunology MeSH
- Interferon-gamma biosynthesis genetics MeSH
- Interleukin-4 biosynthesis genetics MeSH
- Melanoma, Experimental genetics immunology metabolism MeSH
- Mice, Inbred C57BL MeSH
- Mice MeSH
- Cell Line, Tumor MeSH
- Up-Regulation drug effects MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- MeSH
- B7-1 Antigen therapeutic use MeSH
- Immunotherapy methods MeSH
- Melanoma immunology therapy MeSH
- Publication type
- Newspaper Article MeSH
Allogeneic hematopoetic stem cell transplantation (HSCT) represents a unique opportunity to monitor the kinetics of reconstitution of dendritic cells (DCs) and their dynamics in distinct pathologies. We analyzed DCs reconstitution after myeloablative HSCT. We separately analyzed patients with acute GVHD. DCs were monitored from the earliest phase of hematopoetic reconstitution until day +365. Both myeloid DCs and plasmacytoid DCs appeared at earliest stages after engraftment and relative numbers within white blood cells compartment peaked between days 19-25 after HSCT. Their proportion then gradually declined and absolute numbers of both DC subsets remained lower than in controls during the whole follow-up. Patients with acute GVHD had significantly lower numbers of circulating DCs. Decrease in DC counts preceded onset of clinical symptoms by at least 24 h and was independent of corticosteroids administration. This study reveals quantification of plasmacytoid and myeloid DCs as a potential biomarker for the prediction of acute GVHD development.
- MeSH
- B7-1 Antigen biosynthesis immunology MeSH
- B7-2 Antigen biosynthesis immunology MeSH
- Antigens, CD biosynthesis immunology MeSH
- Dendritic Cells cytology immunology MeSH
- Child MeSH
- Adult MeSH
- Financing, Organized MeSH
- Immunophenotyping MeSH
- Immunoglobulins biosynthesis immunology MeSH
- Cohort Studies MeSH
- Humans MeSH
- Membrane Glycoproteins biosynthesis immunology MeSH
- Adolescent MeSH
- Young Adult MeSH
- Graft vs Host Disease immunology pathology MeSH
- Cell Count MeSH
- Child, Preschool MeSH
- Prospective Studies MeSH
- Flow Cytometry MeSH
- Hematopoietic Stem Cell Transplantation methods MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Male MeSH
- Child, Preschool MeSH
- Female MeSH
