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1.
Theranostics ; 12(2): 842-858, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34976216

RESUMO

Background: FcγRIIB, the sole inhibitory receptor of the Fc gamma receptor family, plays pivotal roles in innate and adaptive immune responses. However, the expression and function of FcγRIIB in myeloid-derived suppressor cells (MDSCs) remains unknown. This study aimed to investigate whether and how FcγRIIB regulates the immunosuppressive activity of MDSCs during cancer development. Methods: The MC38 and B16-F10 tumor-bearing mouse models were established to investigate the role of FcγRIIB during tumor progression. FcγRIIB-deficient mice, adoptive cell transfer, mRNA-sequencing and flow cytometry analysis were used to assess the role of FcγRIIB on immunosuppressive activity and differentiation of MDSCs. Results: Here we show that FcγRIIB was upregulated in tumor-infiltrated MDSCs. FcγRIIB-deficient mice showed decreased accumulation of MDSCs in the tumor microenvironment (TME) compared with wild-type mice. FcγRIIB was required for the differentiation and immunosuppressive activity of MDSCs. Mechanistically, tumor cell-derived granulocyte-macrophage colony stimulating factor (GM-CSF) increased the expression of FcγRIIB on hematopoietic progenitor cells (HPCs) by activating specificity protein 1 (Sp1), subsequently FcγRIIB promoted the generation of MDSCs from HPCs via Stat3 signaling. Furthermore, blockade of Sp1 dampened MDSC differentiation and infiltration in the TME and enhanced the anti-tumor therapeutic efficacy of gemcitabine. Conclusion: These results uncover an unrecognized regulatory role of the FcγRIIB in abnormal differentiation of MDSCs during cancer development and suggest a potential therapeutic target for anti-tumor therapy.


Assuntos
Carcinogênese , Diferenciação Celular , Células Supressoras Mieloides/citologia , Receptores de IgG/fisiologia , Evasão Tumoral , Adulto , Animais , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células Supressoras Mieloides/imunologia , Receptores de IgG/deficiência , Receptores de IgG/metabolismo , Transdução de Sinais
2.
Mediators Inflamm ; 2021: 6687250, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34899053

RESUMO

BACKGROUND: C-reactive protein (CRP) is an independent biomarker of systemic inflammation and a predictor of future cardiovascular disease (CVD). More than just a pure bystander, CRP directly interacts with endothelial cells to decrease endothelial nitric oxide synthase (eNOS) expression and bioactivity, decrease nitric oxide (NO) production, and increase the release of vasoconstrictors and adhesion molecules. Race is significantly associated with CRP levels and CVD risks. With aerobic exercise, the vessel wall is exposed to chronic high laminar shear stress (HiLSS) that shifts the endothelium phenotype towards an anti-inflammatory, antioxidant, antiapoptotic, and antiproliferative environment. Thus, the purpose of this study was to assess the racial differences concerning the CRP-induced effects in endothelial cells and the potential role of HiLSS in mitigating these differences. METHODS: Human umbilical vein endothelial cells (HUVECs) from four African American (AA) and four Caucasian (CA) donors were cultured and incubated under the following conditions: (1) static control, (2) CRP (10 µg/mL, 24 hours), (3) CRP receptor (FcγRIIB) inhibitor followed by CRP stimulation, (4) HiLSS (20 dyne/cm2, 24 hours), and (5) HiLSS followed by CRP stimulation. RESULTS: AA HUVECs had significantly higher FcγRIIB receptor expression under both basal and CRP incubation conditions. Blocking FcγRIIB receptor significantly attenuated the CRP-induced decrements in eNOS expression only in AA HUVECs. Finally, HiLSS significantly counteracted CRP-induced effects. CONCLUSION: Understanding potential racial differences in endothelial function is important to improve CVD prevention. Our results shed light on FcγRIIB receptor as a potential contributor to racial differences in endothelial function in AA.


Assuntos
Proteína C-Reativa/farmacologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Afro-Americanos , Doenças Cardiovasculares/prevenção & controle , Células Cultivadas , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Óxido Nítrico Sintase Tipo III/biossíntese , Receptores de IgG/análise , Receptores de IgG/fisiologia , Estresse Mecânico
3.
Front Immunol ; 12: 707542, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34970253

RESUMO

Chimeric antigen receptors (CARs) are fusion proteins with an extracellular antigen recognition domain and numerous intracellular signaling domains that have been genetically modified. CAR-engineered T lymphocyte-based therapies have shown great success against blood cancers; however, potential fatal toxicity, such as in cytokine release syndrome, and high costs are some shortcomings that limit the clinical application of CAR-engineered T lymphocytes and remain to overcome. Natural killer (NK) cells are the focal point of current immunological research owing to their receptors that prove to be promising immunotherapeutic candidates for treating cancer. However, to date, manipulation of NK cells to treat malignancies has been moderately successful. Recent progress in the biology of NK cell receptors has greatly transformed our understanding of how NK cells recognize and kill tumor and infected cells. CAR-NK cells may serve as an alternative candidate for retargeting cancer because of their unique recognition mechanisms, powerful cytotoxic effects especially on cancer cells in both CAR-dependent and CAR-independent manners and clinical safety. Moreover, NK cells can serve as an 'off-the-shelf product' because NK cells from allogeneic sources can also be used in immunotherapies owing to their reduced risk of alloreactivity. Although ongoing fundamental research is in the beginning stages, this review provides an overview of recent developments implemented to design CAR constructs to stimulate NK activation and manipulate NK receptors for improving the efficiency of immunotherapy against cancer, summarizes the preclinical and clinical advances of CAR-NK cells against both hematological malignancies and solid tumors and confronts current challenges and obstacles of their applications. In addition, this review provides insights into prospective novel approaches that further enhance the efficiency of CAR-NK therapies and highlights potential questions that require to be addressed in the future.


Assuntos
Imunoterapia Adotiva/métodos , Células Matadoras Naturais/imunologia , Receptores de Antígenos Quiméricos/imunologia , Receptores de Células Matadoras Naturais/imunologia , Citotoxicidade Celular Dependente de Anticorpos , Apoptose , Ensaios Clínicos como Assunto , Citocinas/fisiologia , Citotoxicidade Imunológica , Desenho de Fármacos , Proteína Ligante Fas/fisiologia , Previsões , Proteínas Ligadas por GPI/fisiologia , Antígenos HLA/imunologia , Humanos , Células Matadoras Naturais/química , Células Matadoras Naturais/transplante , Lentivirus/genética , Ligantes , Macrófagos/imunologia , Subfamília K de Receptores Semelhantes a Lectina de Células NK/fisiologia , Neoplasias/terapia , Receptores de Antígenos Quiméricos/genética , Receptores de IgG/fisiologia , Receptores de Células Matadoras Naturais/classificação , Tolerância a Antígenos Próprios , Subpopulações de Linfócitos T/imunologia , Transdução Genética , Microambiente Tumoral , Receptor fas/fisiologia
4.
Sci Rep ; 11(1): 8383, 2021 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-33863984

RESUMO

Evaluation of the binding and uptake of an antibody in liver non-parenchymal cells (NPC), including liver sinusoidal endothelial cells, is important for revealing its pharmacokinetic (PK) behavior, since NPC has important roles in eliminating an antibody from the blood via the Fc fragment of IgG receptor IIB (FcγRIIB). However, there is currently no in vitro quantitative assay using NPC. This study reports on the development of a cell-based assay for evaluating the binding and uptake of such an antibody using liver NPC of mice and monkeys. In mice, the FcγRIIB-expressing cells were identified in the CD146-positive and CD45-negative fraction by flow cytometry. A titration assay was performed to determine the PK parameters, and the obtained parameter was comparable to that determined by the fitting of the in vivo PK. This approach was also extended to NPC from monkeys. The concentration-dependent binding and uptake was measured to determine the PK parameters using monkey NPC, the FcγRIIB-expressing fraction of which was identified by CD31 and CD45. The findings presented herein demonstrate that the in vitro liver NPC assay using flow cytometry is a useful tool to determine the binding and uptake of biologics and to predict the PK.


Assuntos
Anticorpos Monoclonais/metabolismo , Células Endoteliais/metabolismo , Hepatócitos/metabolismo , Fragmentos Fc das Imunoglobulinas/metabolismo , Receptores de IgG/fisiologia , Animais , Anticorpos Monoclonais/imunologia , Células Endoteliais/imunologia , Haplorrinos , Hepatócitos/imunologia , Fragmentos Fc das Imunoglobulinas/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores de IgG/imunologia
5.
Front Immunol ; 12: 627535, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33790898

RESUMO

The expression of Triggering Receptor Expressed on Myeloid cells (TREM)-1 has been described as a predictive marker for anti-Tumor Necrosis Factor (TNF)-α monoclonal antibody (mAb) therapy responsiveness in patients with inflammatory bowel disease (IBD). Here we investigated expression of TREM-1 specifically in CD14+ monocytes in relation to anti-TNF response. The pretreatment TREM-1 expression levels of CD14+ monocytes of Crohn's disease (CD) patients were predictive of outcome to anti-TNF mAb therapy, with low TREM-1 expression associated with response to anti-TNF. FACSorting of CD14+ monocytes with different TREM-1 levels showed that differentiation towards regulatory CD206+ M2 type macrophages by anti-TNF was suppressed in CD14+ monocytes with high TREM-1 expression. Activity of the Fcγ-Receptor and autophagy pathway, both necessary for M2 type differentiation and the response to anti-TNF, were decreased in CD14+ monocytes with high expression of TREM-1. We confirmed that the activity of the Fcγ-Receptor pathway was decreased in the CD patients that did not respond to anti-TNF therapy and that it was negatively correlated with TREM-1 expression levels in the CD patient cohort. In conclusion, our results indicate that TREM-1 expression levels in CD14+ monocytes associate with decreased autophagy and FcγR activity resulting in decreased differentiation to M2 type regulatory macrophages upon anti-TNF mAb treatment, which may explain anti-TNF non-response in IBD patients with high expression levels of TREM-1.


Assuntos
Autofagia/fisiologia , Doenças Inflamatórias Intestinais/tratamento farmacológico , Monócitos/química , Receptores de IgG/fisiologia , Receptor Gatilho 1 Expresso em Células Mieloides/análise , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adulto , Anticorpos Monoclonais/uso terapêutico , Diferenciação Celular , Feminino , Humanos , Doenças Inflamatórias Intestinais/imunologia , Receptores de Lipopolissacarídeos/análise , Macrófagos/citologia , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/fisiologia
6.
Vet Res ; 52(1): 25, 2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33596995

RESUMO

Porcine reproductive and respiratory syndrome virus (PRRSV) is a highly contagious virus that has led to enormous economic loss worldwide because of ineffective prevention and treatment. In view of their minimized size, high target specificity and affinity, nanobodies have been extensively investigated as diagnostic tools and treatments of many diseases. Previously, a PRRSV Nsp9-specific nanobody (Nb6) was identified as a PRRSV replication inhibitor. When it was fused with cell-penetrating peptide (CPP) TAT, Nb6-TAT could enter the cells for PRRSV suppression. However, delivery of molecules by CPP lack cell specificity and have a short duration of action. PRRSV has a tropism for monocyte/macrophage lineage, which expresses high levels of Fcγ receptors. Herein, we designed a nanobody containing porcine IgG Fc (Fcγ) to inhibit PRRSV replication in PRRSV permissive cells. Fcγ fused Nb6 chimeric antibody (Nb6-pFc) was assembled into a dimer with interchain disulfide bonds and expressed in a Pichia pastoris system. The results show that Nb6-pFc exhibits a well-binding ability to recombinant Nsp9 or PRRSV-encoded Nsp9 and that FcγR-mediated endocytosis of Nb6-pFc into porcine alveolar macrophages (PAM) was in a dose-dependent manner. Nb6-pFc can inhibit PRRSV infection efficiently not only by binding with Nsp9 but also by upregulating proinflammatory cytokine production in PAM. Together, this study proposes the design of a porcine IgG Fc-fused nanobody that can enter PRRSV susceptible PAM via FcγR-mediated endocytosis and inhibit PRRSV replication. This research reveals that nanobody-Fcγ chimeric antibodies might be effective for the control and prevention of monocyte/macrophage lineage susceptible pathogeneses.


Assuntos
Imunoglobulina G/imunologia , Macrófagos Alveolares/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Receptores de IgG/fisiologia , Anticorpos de Domínio Único/imunologia , Proteínas não Estruturais Virais/imunologia , Animais , Síndrome Respiratória e Reprodutiva Suína/imunologia , Síndrome Respiratória e Reprodutiva Suína/prevenção & controle , Síndrome Respiratória e Reprodutiva Suína/virologia , Vírus da Síndrome Respiratória e Reprodutiva Suína/fisiologia , Anticorpos de Domínio Único/química , Suínos , Replicação Viral
7.
Front Immunol ; 11: 2128, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33123121

RESUMO

Tumor cells develop various mechanisms to escape immune surveillance. In this context, oncometabolites secreted by tumor cells due to deregulated metabolic pathways, have been in the spotlight of researchers during the last years. 5'-Deoxy-5'-methylthioadenosine (MTA) phosphorylase (MTAP) deficiency in tumors results in the accumulation of MTA within the tumor microenvironment and thereby negatively influencing immune functions of various immune cells, including T and NK cells. The influence of MTA on T cell activation has been recently described in more detail, while its impact on NK cells is still largely unknown. Therefore, we aimed to illuminate the molecular mechanism of MTA-induced NK cell dysfunction. NK cell cytotoxicity against target cells was reduced in the presence of MTA in a dose-dependent manner, while NK cell viability remained unaffected. Furthermore, we revealed that MTA blocks NK cell degranulation and cytokine production upon target cell engagement as well as upon antibody stimulation. Interestingly, the immune-suppressive effect of MTA was less pronounced in healthy donors harboring an expansion of NKG2C+ NK cells. Finally, we demonstrated that MTA interferes with various signaling pathways downstream of the CD16 receptor upon NK cell activation, including the PI3K/AKT/S6, MAPK/ERK, and NF-κB pathways. In summary, we revealed that MTA blocks NK cell functions like cytotoxicity and cytokine production by interfering with the signaling cascade of activating NK cell receptors. Specific targeting of MTA metabolism in MTAP-deficient tumors therefore could offer a promising new strategy to reverse immune dysfunction of NK cells within the tumor microenvironment.


Assuntos
Desoxiadenosinas/farmacologia , Células Matadoras Naturais/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , Purina-Núcleosídeo Fosforilase/metabolismo , Transdução de Sinais/efeitos dos fármacos , Tionucleosídeos/farmacologia , Antígenos CD57/imunologia , Degranulação Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/biossíntese , Citotoxicidade Imunológica , Proteínas Ligadas por GPI/fisiologia , Humanos , Interferon gama/biossíntese , Interferon gama/genética , Células K562 , Células Matadoras Naturais/imunologia , Subpopulações de Linfócitos/efeitos dos fármacos , Subpopulações de Linfócitos/imunologia , Proteína 1 de Membrana Associada ao Lisossomo/biossíntese , Proteína 1 de Membrana Associada ao Lisossomo/genética , NF-kappa B/fisiologia , Subfamília C de Receptores Semelhantes a Lectina de Células NK/análise , Proteína-Arginina N-Metiltransferases/antagonistas & inibidores , Receptores de IgG/fisiologia , Evasão Tumoral , Ensaio Tumoral de Célula-Tronco
8.
Front Immunol ; 11: 1097, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32595635

RESUMO

Phagocytosis is a specialized process that enables cellular ingestion and clearance of microbes, dead cells and tissue debris that are too large for other endocytic routes. As such, it is an essential component of tissue homeostasis and the innate immune response, and also provides a link to the adaptive immune response. However, ingestion of large particulate materials represents a monumental task for phagocytic cells. It requires profound reorganization of the cell morphology around the target in a controlled manner, which is limited by biophysical constraints. Experimental and theoretical studies have identified critical aspects associated with the interconnected biophysical properties of the receptors, the membrane, and the actin cytoskeleton that can determine the success of large particle internalization. In this review, we will discuss the major physical constraints involved in the formation of a phagosome. Focusing on two of the most-studied types of phagocytic receptors, the Fcγ receptors and the complement receptor 3 (αMß2 integrin), we will describe the complex molecular mechanisms employed by phagocytes to overcome these physical constraints.


Assuntos
Fagocitose/imunologia , Fagocitose/fisiologia , Citoesqueleto de Actina/metabolismo , Animais , Fenômenos Biofísicos , Movimento Celular/imunologia , Movimento Celular/fisiologia , Extensões da Superfície Celular/imunologia , Extensões da Superfície Celular/fisiologia , Humanos , Ligantes , Antígeno de Macrófago 1/química , Antígeno de Macrófago 1/imunologia , Antígeno de Macrófago 1/fisiologia , Modelos Imunológicos , Miosina Tipo II/imunologia , Miosina Tipo II/fisiologia , Fagossomos/imunologia , Fagossomos/fisiologia , Conformação Proteica , Pseudópodes/imunologia , Pseudópodes/fisiologia , Receptores de IgG/química , Receptores de IgG/imunologia , Receptores de IgG/fisiologia
9.
Cancer Cell ; 37(6): 850-866.e7, 2020 06 08.
Artigo em Inglês | MEDLINE | ID: mdl-32442402

RESUMO

Anti-CD40 monoclonal antibodies (mAbs) comprise agonists and antagonists, which display promising therapeutic activities in cancer and autoimmunity, respectively. We previously showed that epitope and isotype interact to deliver optimal agonistic anti-CD40 mAbs. The impact of Fc engineering on antagonists, however, remains largely unexplored. Here, we show that clinically relevant antagonists used for treating autoimmune conditions can be converted into potent FcγR-independent agonists with remarkable antitumor activity by isotype switching to hIgG2. One antagonist is converted to a super-agonist with greater potency than previously reported highly agonistic anti-CD40 mAbs. Such conversion is dependent on the unique disulfide bonding properties of the hIgG2 hinge. This investigation highlights the transformative capacity of the hIgG2 isotype for converting antagonists to agonists to treat cancer.


Assuntos
Anticorpos Monoclonais/farmacologia , Antígenos CD40/imunologia , Ligante de CD40/imunologia , Células Dendríticas/imunologia , Switching de Imunoglobulina/imunologia , Imunoglobulina G/imunologia , Neoplasias/tratamento farmacológico , Animais , Anticorpos Monoclonais/imunologia , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/imunologia , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Células Dendríticas/efeitos dos fármacos , Switching de Imunoglobulina/genética , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neoplasias/imunologia , Neoplasias/metabolismo , Neoplasias/patologia , Receptores de IgE/fisiologia , Receptores de IgG/fisiologia , Neoplasias do Timo/tratamento farmacológico , Neoplasias do Timo/imunologia , Neoplasias do Timo/metabolismo , Neoplasias do Timo/patologia
10.
Mol Cancer Ther ; 19(4): 1040-1051, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31974274

RESUMO

CD137 (TNFRSF9, 4-1BB) agonist antibodies (mAb) have demonstrated potent antitumor activity with memory response while causing hepatotoxicity in mouse models. In clinical trials, the degrees of liver toxicity of anti-CD137 vary from grade 4 transaminitis (urelumab) to nonexistent (utomilumab). To exploit the antitumor potential of CD137 signaling, we identified a new class of CD137 agonist mAbs with strong antitumor potency without significant transaminitis in vivo compared with CD137 agonists previously reported. These mAbs are cross-reactive to mouse and cynomolgus monkey and showed cross-linking-dependent T-cell costimulation activity in vitro Antitumor efficacy was maintained in Fc gamma receptor (FcγR) III-deficient mice but diminished in FcγRIIB-deficient mice, suggesting the critical role for FcγRIIB to provide cross-linking in vivo Interestingly, a single dose of an affinity-reduced variant was sufficient to control tumor growth, but a higher affinity variant did not improve efficacy. These observations suggest that binding epitope and FcγR interaction, but not necessarily high affinity, are important for antitumor efficacy and reduced liver toxicity of CD137 mAb. Our study suggests the possibility of CD137 agonist therapy with improved safety profile in humans.


Assuntos
Anticorpos Monoclonais/farmacologia , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Neoplasias do Colo/tratamento farmacológico , Reagentes para Ligações Cruzadas/química , Epitopos/imunologia , Melanoma Experimental/tratamento farmacológico , Receptores de IgG/fisiologia , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral/imunologia , Animais , Apoptose , Proliferação de Células , Neoplasias do Colo/imunologia , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Reagentes para Ligações Cruzadas/metabolismo , Feminino , Humanos , Melanoma Experimental/imunologia , Melanoma Experimental/metabolismo , Melanoma Experimental/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células Tumorais Cultivadas
11.
Neurobiol Aging ; 88: 24-32, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31901293

RESUMO

Transcriptomics has become an important tool for identification of biological pathways dysregulated in Alzheimer's disease (AD). We performed a network-based gene expression analysis of blood-based microarray gene expression profiles using 2 independent cohorts, Alzheimer's Disease Neuroimaging Initiative (ADNI; N = 661) and AddNeuroMed (N = 674). Weighted gene coexpression network analysis identified 17 modules from ADNI and 13 from AddNeuroMed. Four of the modules derived in ADNI were significantly related to AD; 5 modules in AddNeuroMed were significant. Gene-set enrichment analysis of the AD-related modules identified and replicated 3 biological pathways including the Fc gamma receptor-mediated phagocytosis pathway. Module-based association analysis showed the AD-related module, which has the 3 pathways, to be associated with cognitive function and neuroimaging biomarkers. Gene-based association analysis identified PRKCD in the Fc gamma receptor-mediated phagocytosis pathway as being significantly associated with cognitive function and cerebrospinal fluid biomarkers. The identification of the Fc gamma receptor-mediated phagocytosis pathway implicates the peripheral innate immune system in the pathophysiology of AD. PRKCD is known to be related to neurodegeneration induced by amyloid-ß.


Assuntos
Doença de Alzheimer/genética , Perfilação da Expressão Gênica/métodos , Expressão Gênica , Estudos de Associação Genética , Fagocitose/genética , Proteína Quinase C-delta , Receptores de IgG/genética , Receptores de IgG/fisiologia , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Transcriptoma/genética , Doença de Alzheimer/imunologia , Doença de Alzheimer/psicologia , Peptídeos beta-Amiloides/metabolismo , Cognição , Imunidade Inata
12.
Cancer Immunol Immunother ; 68(11): 1769-1778, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31616964

RESUMO

Various immune cells are recruited in the tumor microenvironment. It is well established that cellular immune responses, such as cytotoxic or suppressive activities, play an important role in regulating tumor growth and metastasis. However, the contribution of humoral immune responses against tumors is poorly understood. Fc receptors constitute critical elements for the up- or downregulation of immune responses through immune complexes. Here, we examined the potential role of the inhibitory Fc receptor, Fcγ receptor IIB (FcγRIIB), in tumor immunity using a mouse model. Our findings indicated that tumor-specific antibodies are induced in tumor-bearing mice and control tumor immunity. FcγRIIB deletion significantly improved both cellular and humoral immunity against tumors and delayed tumor growth. These findings indicated that spontaneous antibodies against tumors create a suppressive tumor microenvironment through FcγRIIB signaling, thus suggesting an attractive therapeutic target for cancer immunotherapy.


Assuntos
Anticorpos/imunologia , Carcinoma Pulmonar de Lewis/terapia , Imunoterapia , Receptores de IgG/fisiologia , Timoma/terapia , Neoplasias do Timo/terapia , Microambiente Tumoral/imunologia , Animais , Carcinoma Pulmonar de Lewis/imunologia , Carcinoma Pulmonar de Lewis/metabolismo , Carcinoma Pulmonar de Lewis/patologia , Modelos Animais de Doenças , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores de IgG/imunologia , Timoma/imunologia , Timoma/metabolismo , Timoma/patologia , Neoplasias do Timo/imunologia , Neoplasias do Timo/metabolismo , Neoplasias do Timo/patologia , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
13.
Arterioscler Thromb Vasc Biol ; 39(7): 1379-1389, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31092015

RESUMO

Objective- Investigate the impact of modulating B cell FcγRIIb (Fcγ receptor IIb) expression on atherosclerosis. Approach and Results- Western diet-induced atherosclerosis was assessed in Ldlr-/- or Apoe-/- mice with B cell-specific overexpression of FcγRIIb or with an FcγRIIb promoter mutation that alters FcγRIIb expression in germinal center (GC) B cells. In males, overexpression of FcγRIIb on B cells severely reduced activated, class switched B cell responses, as indicated by reductions in GC B cells, plasma cells, and serum IgG but not IgM antibodies. Male mice overexpressing FcγRIIb developed less atherosclerosis, suggesting a pathogenic role for GC B cell IgG responses. In support of this hypothesis, male mice with a promoter polymorphism-driven reduction in FcγRIIb on GC B cells but not plasma cells have a converse phenotype of enhanced GC responses and IgG2c antibodies and enhanced atherosclerosis. IgG2c significantly enhanced TNF (tumor necrosis factor) secretion by CD11b+ CD11c+ cells expressing the high-affinity receptor FcγRIV. In females, overexpression of FcγRIIb on B cells not only reduced GC B cell responses but also substantially reduced B-1 cells and IgM antibodies, which translated into acceleration of atherosclerosis. Promoter-driven reduction in FcγRIIb did not alter GC B cell responses in females and, therefore, had no impact on atherosclerosis. Conclusions- B cell FcγRIIb differentially alters proatherogenic adaptive GC B cell and atheroprotective innate B-1 responses in male and female mice fed a western diet. Our results highlight the importance of a better understanding and ability to selectively target B cell responses in future immunotherapeutic approaches against human cardiovascular disease. Visual Overview- An online visual overview is available for this article.


Assuntos
Aterosclerose/imunologia , Linfócitos B/imunologia , Centro Germinativo/imunologia , Receptores de IgG/fisiologia , Animais , Apolipoproteínas E/fisiologia , Feminino , Imunidade Inata , Imunoglobulina M/biossíntese , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fator de Necrose Tumoral alfa/biossíntese
14.
Atherosclerosis ; 285: 108-119, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31051414

RESUMO

BACKGROUND AND AIMS: Circulating levels of oxidized lipoprotein (oxLDL) correlate with myocardial infarction risk and atherosclerosis severity. Our previous study demonstrates that oxLDL immune complexes (oxLDL-ICs) can signal through FcγRs on bone marrow-derived dendritic cells (BMDCs) and enhance their activation and inflammatory cytokine secretion. While global FcγR-/- studies have shown that activating FcγRs are proatherogenic, the role of the inhibitory FcγRIIb is unclear. We sought to determine the role of DC-specific FcγRIIb in atherosclerosis. METHODS: Bone marrow chimeras were generated by rescuing lethally irradiated Ldlr-/- mice with hematopoietic cells from littermate CD11c-Cre+ or CD11c-Cre-Fcgr2bfl/fl donors. Four weeks following transplant, recipients were placed on a Western diet for eight weeks. Various tissues and organs were analyzed for differences in inflammation. RESULTS: Quantitation of atherosclerosis in the proximal aorta demonstrated a 58% increase in female CD11c-Cre+Fcgr2bfl/fl recipients, but a surprising 44% decrease in male recipients. Hepatic cholesterol and triglycerides were increased in female CD11c-Cre+Fcgr2bfl/fl recipients. This was associated with an increase in CD36 and MHC Class II expression on hepatic CD11c+CD11b+ DCs in female livers. In contrast, male CD11c-Cre+Fcgr2bfl/fl recipients had decreased hepatic lipids with a corresponding decrease in CD36 and MHC Class II expression on CD11c+ cells. Interestingly, both sexes of CD11c-Cre+Fcgr2bfl/fl recipients had significant decreases in serum cholesterol and TGs with corresponding decreases in liver Fasn transcripts. CONCLUSIONS: The absence of FcγRIIb expression on CD11c+ cells results in sex-dependent alteration in liver inflammation influencing atherogenesis and sex-independent modulation of serum cholesterol and TGs.


Assuntos
Aterosclerose/sangue , Colesterol/sangue , Receptores de IgG/fisiologia , Triglicerídeos/sangue , Animais , Antígenos CD11/biossíntese , Células Dendríticas/metabolismo , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptores de IgG/biossíntese , Fatores Sexuais
15.
Front Immunol ; 10: 252, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30858848

RESUMO

Activating Fcγ receptors associated with Fc receptor γ-chain (FcRγ) are critical for mediating neutrophil effector functions in immune complex-mediated autoimmune diseases. FcRγ contains ITAM tyrosines and the in vivo role of these tyrosines has not been defined in neutrophils and arthritis. In this study, the in vivo functions of FcRγ ITAM tyrosines were characterized using wild type and ITAM tyrosine mutant (Y65F/Y76F) transgenic mice crossed to an FcRγ-deficient genetic background. FcRγ-deficient neutrophils showed undetectable cell surface expression of the activating Fcγ receptor IV, defective immune complex-induced superoxide production, degranulation and spreading. Although the re-expression of both the wild type and the ITAM tyrosine mutant (Y65F/Y76F) FcRγ could restore activating Fcγ receptor expression of FcRγ-deficient neutrophils, only the wild type transgenic form could mediate Fcγ receptor-dependent effector functions. In contrast, neutrophils carrying ITAM tyrosine mutant FcRγ were unable to produce superoxide, mediate degranulation and perform active spreading. In addition, our results confirmed the protection of FcRγ-deficient mice from autoimmune arthritis. Importantly, the presence of the wild type FcRγ transgene, in contrast to the ITAM tyrosine mutant transgene, partially reversed autoimmune arthritis development. The reversing effect of the wild type transgene was even more robust when animals carried the wild type transgene in a homozygous form. Collectively, FcRγ ITAM tyrosines play a critical role in the induction of neutrophil effector responses, the initiation and progression of an autoantibody-induced experimental arthritis in vivo, indicating a signaling, rather than just a receptor stabilizing function of the molecule.


Assuntos
Artrite Experimental/etiologia , Ativação de Neutrófilo , Receptores de IgG/fisiologia , Motivos de Aminoácidos , Animais , Complexo Antígeno-Anticorpo/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Receptores de IgG/química , Relação Estrutura-Atividade , Tirosina/fisiologia
16.
Cancer Cell ; 34(2): 331-345.e11, 2018 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-30107179

RESUMO

Therapeutic antibodies targeting ovarian cancer (OvCa)-enriched receptors have largely been disappointing due to limited tumor-specific antibody-dependent cellular cytotoxicity. Here we report a symbiotic approach that is highly selective and superior compared with investigational clinical antibodies. This bispecific-anchored cytotoxicity activator antibody is rationally designed to instigate "cis" and "trans" cytotoxicity by combining specificities against folate receptor alpha-1 (FOLR1) and death receptor 5 (DR5). Whereas the in vivo agonist DR5 signaling requires FcγRIIB interaction, the FOLR1 anchor functions as a primary clustering point to retain and maintain a high level of tumor-specific apoptosis. The presented proof of concept study strategically makes use of a tumor cell-enriched anchor receptor for agonist death receptor targeting to potentially generate a clinically viable strategy for OvCa.


Assuntos
Anticorpos Biespecíficos/uso terapêutico , Receptor 1 de Folato/antagonistas & inibidores , Neoplasias Ovarianas/tratamento farmacológico , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/antagonistas & inibidores , Animais , Linhagem Celular Tumoral , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Ovarianas/patologia , Receptores de IgG/fisiologia
17.
Cell Syst ; 7(1): 41-48.e5, 2018 07 25.
Artigo em Inglês | MEDLINE | ID: mdl-29960887

RESUMO

Many immune receptors transduce activation across the plasma membrane through their clustering. With Fcγ receptors (FcγRs), this clustering is driven by binding to antibodies of differing affinities that are in turn bound to multivalent antigen. As a consequence of this activation mechanism, accounting for and rationally manipulating immunoglobulin (Ig)G effector function is complicated by, among other factors, differing affinities between FcγR species and changes in the valency of antigen binding. In this study, we show that a model of multivalent receptor-ligand binding can effectively account for the contribution of IgG-FcγR affinity and immune complex valency. This model in turn enables us to make specific predictions about the effect of immune complexes of defined composition. In total, these results enable both rational immune complex design for a desired IgG effector function and the deconvolution of effector function by immune complexes.


Assuntos
Fragmentos Fc das Imunoglobulinas/química , Receptores de IgG/metabolismo , Animais , Anticorpos/metabolismo , Anticorpos/fisiologia , Anticorpos Monoclonais/imunologia , Afinidade de Anticorpos , Complexo Antígeno-Anticorpo/imunologia , Subpopulações de Linfócitos B/metabolismo , Células CHO , Proteínas de Transporte/metabolismo , Membrana Celular/fisiologia , Cricetulus , Humanos , Fenômenos do Sistema Imunológico/fisiologia , Fragmentos Fc das Imunoglobulinas/fisiologia , Imunoglobulina G/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Ligação Proteica/fisiologia , Receptores de IgG/fisiologia
18.
Clin Exp Rheumatol ; 36(2): 284-293, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29352854

RESUMO

OBJECTIVES: Anti-citrullinated protein antibodies (ACPA) are highly specific for rheumatoid arthritis (RA). Here, we studied binding of ACPA-IgG immune complexes (IC) to individual Fc gamma receptors (FcγR) to identify potential effector mechanisms by which ACPA could contribute to RA pathogenesis. METHODS: ACPA-IgG1 and control IgG1(IgG1 depleted of ACPA-IgG1) were isolated from plasma and synovial fluid (SF) of RA patients by affinity chromatography using CCP2 peptides. Subsequently, IC were generated using fluorescently labelled F(ab')2 fragments against the F(ab')2 region of IgG, or by using citrullinated fibrinogen. IC were incubated with FcγR-transfected CHO cell lines or neutrophils from healthy donors. FcγR binding of IC was analysed by flow cytometry in the presence or absence of specific blocking antibodies. RESULTS: ACPA-IgG1 IC predominantly bound to FcγRI and FcγRIIIA on FcγR-transfected CHO cell lines, while much lower binding was observed to FcγRIIA and FcγRIIB. ACPA-IgG1 IC showed reduced binding to FcγRIIIA compared to control IgG1 IC, in line with enhanced ACPA-IgG1 Fc core-fucosylation. Neutrophils activated in vitro to induce de novo expression of FcγRI showed binding of ACPA-IgG IC, and blocking studies revealed that almost 30% of ACPA-IgG IC binding to activated neutrophils was mediated by FcγRI. CONCLUSIONS: Our studies show that ACPA-IgG1 IC bind predominately to activating FcγRI and FcγRIIIA, and highlight FcγRI expressed by activated neutrophils as relevant receptor for these IC. As neutrophils isolated from SF exhibit an activated state and express FcγRI in the synovial compartment, this IC-binding could contribute to driving disease pathogenesis in RA.


Assuntos
Anticorpos Anti-Proteína Citrulinada/metabolismo , Complexo Antígeno-Anticorpo/metabolismo , Artrite Reumatoide/etiologia , Receptores de IgG/metabolismo , Idoso , Artrite Reumatoide/imunologia , Feminino , Glicosilação , Humanos , Masculino , Pessoa de Meia-Idade , Neutrófilos/fisiologia , Receptores de IgG/fisiologia , Sinovite/etiologia
19.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 33(6): 746-750, 2017 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-28615096

RESUMO

Objective To investigate the role of IgG Fc receptorI (FcγRI) in lipopolysaccharide (LPS)-induced apoptosis of the rat PC12 cells. Methods PC12 cells were treated with different concentrations of LPS (50, 125, 250, 500, 1000 µg/mL) for 24 hours and cell viability was analyzed by MTT assay. The appropriate concentration of LPS (500 µg/mL) was chosen for the following experiments. PC12 cells in the logarithmic growth phase were divided randomly into three groups: the control group without LPS, the 500 µg/mL LPS treated group and the 500 µg/mL LPS plus 0.2 µg/mL FcγRI neutralizer group. After24-hour different treatments, the mRNA and protein levels of FcγRIwere detected by quantitative real-time PCR and Western blotting, respectively. The apoptosis rate of PC12 cells was determined by flow cytometry combined with annexinV-FITC/PI double staining. The protein expression levels of caspase-3, Bcl-2 and BAX were measured by immunohistochemistry. Results PC12 cell viability decreased in a LPS dose-dependent manner. Compared to the control group, the protein and mRNA expression of FcγRI were upregulated, the expression levels of caspase-3, Bcl-2 and BAX proteins were elevated, and the apoptosis rate of PC12 cells was raised as well in the LPS treated group. Compared to the LPS treated group, the protein and mRNA levels of FcγRI were significantly lower along with significantly reduced expressions of Caspase-3 and BAX and inhibited cell apoptosis in the FcγRIneutralizer treated group, while Bcl-2 protein expression was upregulated. Conclusion FcγRIis involved in the LPS-induced apoptosis in PC12 cells.


Assuntos
Apoptose/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Receptores de IgG/fisiologia , Animais , Caspase 3/metabolismo , Células PC12 , Proteínas Proto-Oncogênicas c-bcl-2/análise , Ratos , Receptores de IgG/análise , Receptores de IgG/genética , Proteína X Associada a bcl-2/análise
20.
J Exp Med ; 214(5): 1509-1528, 2017 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-28389502

RESUMO

Dendritic cells (DCs) are efficient antigen-presenting cells equipped with various cell surface receptors for the direct or indirect recognition of pathogenic microorganisms. Interestingly, not much is known about the specific expression pattern and function of the individual activating and inhibitory Fcγ receptors (FcγRs) on splenic DC subsets in vivo and how they contribute to the initiation of T cell responses. By targeting antigens to select activating and the inhibitory FcγR in vivo, we show that antigen uptake under steady-state conditions results in a short-term expansion of antigen-specific T cells, whereas under inflammatory conditions especially, the activating FcγRIV is able to induce superior CD4+ and CD8+ T cell responses. Of note, this effect was independent of FcγR intrinsic activating signaling pathways. Moreover, despite the expression of FcγRIV on both conventional splenic DC subsets, the induction of CD8+ T cell responses was largely dependent on CD11c+CD8+ DCs, whereas CD11c+CD8- DCs were critical for priming CD4+ T cell responses.


Assuntos
Células Dendríticas/fisiologia , Receptores de IgG/fisiologia , Linfócitos T/fisiologia , Animais , Endocitose/fisiologia , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Receptores de Antígenos de Linfócitos T/fisiologia
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