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1.
Fish Shellfish Immunol ; 93: 801-814, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31419534

RESUMO

The signaling mediated by small non-proteinogenic molecules, which probably have the capacity to serve as a bridge amongst complex systems is one of the most exiting challenges for the study. In the current report, stem cells differentiation of the immune system in Nile tilapia treated with sub-basal doses of GABA evaluated as c-kit+ and Sca-1+ cells disappearance on pronephros, thymus, spleen and peripheral blood mononuclear cells by flow cytometry was assessed. Explanation of biological response was performed by molecular docking approach and multiparametric analysis. Stem cell differentiation depends on a delicate balance of negative and positive interactions of this neurotransmitter with receptors and transcription factors involved in this process. This in turn depends on the type of interaction with hematopoietic niche to differentiate into primordial, early or late hematopoiesis as well as from the dose delivery. In fish treated with the low doses of GABA (0.1% over basal value) primordial hematopoiesis is regulated by interaction of glutamate (Glu) with the Ly-6 antigen. Early hematopoiesis was influenced by the bond of GABA near or adjacent to turns of FLTR3-Ig-IV domain. During late hematopoiesis, negative regulation by structural modifications on PU.1/IRF-4 complex, IL-7Rα and GM-CSFR mainly prevails. Results of molecular docking were in agreement with the percentages of the main blood cells lineages estimated in pronephros by flow cytometry. Current study provides the first evidences about the role of inhibitory and excitatory neurotransmitters such as GABA and Glu, respectively with the most transcriptional factors and receptors involved on hematopoiesis in adult Nile tilapia.


Assuntos
Ciclídeos/fisiologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Células-Tronco Hematopoéticas/fisiologia , Animais , Antígenos Ly/genética , Antígenos Ly/imunologia , Diferenciação Celular/fisiologia , Ciclídeos/imunologia , Células-Tronco Hematopoéticas/imunologia , Sistema Imunitário/fisiologia , Proteínas Proto-Oncogênicas c-kit/genética , Proteínas Proto-Oncogênicas c-kit/imunologia , Ácido gama-Aminobutírico/farmacologia
2.
Folia Histochem Cytobiol ; 57(2): 94-100, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31237344

RESUMO

INTRODUCTION: A reduced number of interstitial Cajal-like cells (ICLCs) in the gallbladder have been proposed to play a role in the pathogenesis of cholelithiasis. Therefore, this prospective study was conducted to investigate the relationship between gallbladder contractility and the number of gallbladder ICLCs in patients with cholelithiasis. MATERIAL AND METHODS: Patients admitted to the Department of Hepatobiliary Surgery for cholecystectomy were divided into the cholelithiasis (n = 18) and non-cholelithiasis (n = 8) groups based on their clinical data. Patients' clinical data were collected on admission, and B-mode ultrasonography was performed to assess their gallbladder contractility. The resected gallbladder specimens were fixed, paraffin sections mounted on slides, and the immunofluorescence staining with the anti-human CD-117 and anti-human tryptase antibodies was performed to identify ICLSs and mast cells, respectively. The number of ICLCs was counted in 10 high-power fields (HPFs) randomly. RESULTS: Independent sample t-tests revealed differences between the cholelithiasis and non-cholelithiasis groups in the number of ICLCs (mean ± standard deviation: 88.61 ± 28.22 vs. 115.89 ± 27.87 per HPFs, P = 0.032) and gallbladder contractility (43.94% ± 18.50% vs. 61.00% ± 20.50%, P = 0.046). Pearson and Spearman cor-relation analyses revealed no significant correlation between the number of ICLCs and gallbladder contractility. CONCLUSION: The results suggest that the number of gallbladder ICLCs in the wall of the gallbladder of patients with or without cholelithiasis is not a decisive factor affecting gallbladder contractility.


Assuntos
Colelitíase/fisiopatologia , Esvaziamento da Vesícula Biliar/fisiologia , Vesícula Biliar/citologia , Vesícula Biliar/fisiologia , Telócitos/citologia , Adulto , Idoso , Animais , Anticorpos/imunologia , Contagem de Células , Colelitíase/patologia , Feminino , Vesícula Biliar/patologia , Cabras , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Estudos Prospectivos , Proteínas Proto-Oncogênicas c-kit/imunologia , Coelhos , Telócitos/patologia , Triptases/imunologia
3.
Anal Sci ; 35(7): 811-813, 2019 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-30930352

RESUMO

An immunosensor based on surface plasmon resonance was applied to detect mast cells expressing c-Kit. Sufficient detection of the mast cells was achieved by covalent immobilization of gelatin firstly on the sensor surface and followed by covalent binding of the anti-c-Kit antibody to lysine residues in the gelatin molecules through bis(sulfosuccinimidyl)suberate (BS3) treatment. By using BS3, which is a homo-bifunctional reagent, the lysine residues of the anti-c-Kit antibody easily bound to the lysine residues of the gelatin in the physiological condition. The lower limit of detection was 104 cells/mL.


Assuntos
Anticorpos Imobilizados/química , Gelatina/química , Regulação da Expressão Gênica , Imunoensaio/métodos , Mastócitos/citologia , Proteínas Proto-Oncogênicas c-kit/metabolismo , Ressonância de Plasmônio de Superfície/métodos , Animais , Anticorpos Imobilizados/imunologia , Ácidos Decanoicos/química , Humanos , Limite de Detecção , Mastócitos/metabolismo , Camundongos , Proteínas Proto-Oncogênicas c-kit/imunologia , Succinimidas/química
4.
Anal Sci ; 35(2): 223-225, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30745512

RESUMO

An immunosensor based on surface plasmon resonance was developed for detection of c-Kit expressed on a cell surface. The combination of the antibody solution modified with gelatin before immobilization to the sensor chip and its blocking with gelatin drastically decreased the nonspecific reaction. The condition may be useful for the detection of various cells by using antibody against cell surface marker including the c-Kit.


Assuntos
Regulação da Expressão Gênica , Imunoensaio/métodos , Proteínas Proto-Oncogênicas c-kit/metabolismo , Ressonância de Plasmônio de Superfície/métodos , Anticorpos Monoclonais/imunologia , Linhagem Celular Tumoral , Células HEK293 , Humanos , Proteínas Proto-Oncogênicas c-kit/imunologia
5.
Nat Commun ; 10(1): 616, 2019 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-30728353

RESUMO

Hematopoietic chimerism after allogeneic bone marrow transplantation may establish a state of donor antigen-specific tolerance. However, current allotransplantation protocols involve genotoxic conditioning which has harmful side-effects and predisposes to infection and cancer. Here we describe a non-genotoxic conditioning protocol for fully MHC-mismatched bone marrow allotransplantation in mice involving transient immunosuppression and selective depletion of recipient hematopoietic stem cells with a CD117-antibody-drug-conjugate (ADC). This protocol resulted in multilineage, high level (up to 50%), durable, donor-derived hematopoietic chimerism after transplantation of 20 million total bone marrow cells, compared with ≤ 2.1% hematopoietic chimerism from 50 million total bone marrow cells without conditioning. Moreover, long-term survival of bone marrow donor-type but not third party skin allografts is achieved in CD117-ADC-conditioned chimeric mice without chronic immunosuppression. The only observed adverse event is transient elevation of liver enzymes in the first week after conditioning. These results provide proof-of-principle for CD117-ADC as a non-genotoxic, highly-targeted conditioning agent in allotransplantation and tolerance protocols.


Assuntos
Transplante de Medula Óssea/métodos , Sobrevivência de Enxerto , Transplante de Células-Tronco Hematopoéticas/métodos , Imunoconjugados/farmacologia , Proteínas Proto-Oncogênicas c-kit/imunologia , Tolerância ao Transplante/efeitos dos fármacos , Animais , Células da Medula Óssea/efeitos dos fármacos , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/prevenção & controle , Células-Tronco Hematopoéticas , Tolerância Imunológica , Imunossupressão/métodos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Modelos Animais , Pele/patologia , Transplante de Pele/métodos , Quimeras de Transplante , Transplante Homólogo
6.
Nat Commun ; 10(1): 617, 2019 02 06.
Artigo em Inglês | MEDLINE | ID: mdl-30728354

RESUMO

Hematopoietic stem cell transplantation (HSCT) is a curative therapy for blood and immune diseases with potential for many settings beyond current standard-of-care. Broad HSCT application is currently precluded largely due to morbidity and mortality associated with genotoxic irradiation or chemotherapy conditioning. Here we show that a single dose of a CD117-antibody-drug-conjugate (CD117-ADC) to saporin leads to > 99% depletion of host HSCs, enabling rapid and efficient donor hematopoietic cell engraftment. Importantly, CD117-ADC selectively targets hematopoietic stem cells yet does not cause clinically significant side-effects. Blood counts and immune cell function are preserved following CD117-ADC treatment, with effective responses by recipients to both viral and fungal challenges. These results suggest that CD117-ADC-mediated HSCT pre-treatment could serve as a non-myeloablative conditioning strategy for the treatment of a wide range of non-malignant and malignant diseases, and might be especially suited to gene therapy and gene editing settings in which preservation of immunity is desired.


Assuntos
Transplante de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas/efeitos dos fármacos , Imunoconjugados/farmacologia , Proteínas Proto-Oncogênicas c-kit/imunologia , Animais , Medula Óssea/efeitos dos fármacos , Transplante de Medula Óssea , Candida albicans/patogenicidade , Morte Celular , Linhagem Celular , Feminino , Terapia Genética , Humanos , Imunoconjugados/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias , Doadores de Tecidos
8.
Acta Chir Orthop Traumatol Cech ; 85(5): 351-358, 2018.
Artigo em Tcheco | MEDLINE | ID: mdl-30383532

RESUMO

PURPOSE OF THE STUDY This study deals with the possibilities and application of immunohistochemical methods to detect mast and dendritic cells in periprosthetic tissues in patients with aseptically loosened total joint replacements of the knee and hip. The purpose of the study was to quantify and characterize the distribution of mast and dendritic cells in the examined samples and to study the statistically significant relations between the aforementioned cell populations and selected parameters characterizing the patients, implants or tissue response. Based on the proved findings, a possible relation between mast and dendritic cells and histomorphological patterns of aseptic loosening and the benefit of the applied immunohistochemical methods was evaluated. MATERIAL AND METHODS Periprosthetic tissues from a total of 31 patients (17 patients after a revision surgery of hip prosthesis, 14 patients after a revision surgery of knee prosthesis) were examined. The collected samples were processed according to the standard protocol for the purposes of histological and immunochemical examination. Antibodies against tryptase and CD117 were used for immunohistochemical detection of mast cells. Dendritic cells were detected by means of S100 and CD1a antibodies. Quantification of both the cell populations was carried out by optical microscopy in 20 high power fields at 400-times magnification. From among the applied methods we picked the more sensitive one for statistical evaluation. It was tryptase in the case of mast cells and S100 in the case of dendritic cells. RESULTS Mast and dendritic cells were mostly distributed dispersively in periprosthetic tissues; however, they also occurred in groups perivasally or near necrotic parts. The examined samples showed the presence of 60 mast cells and 50 dendritic cells on average. The increased density of mast and dendritic cells was associated with polypously formed pseudosynovium and cement fixation of prostheses; this relation was statistically significant. It was impossible to prove the correlation between the quantity of the observed cell populations and the nature and the number of the observed particles because wear particles were present dispersely in all the samples. Another statistically significant relation to the type of material or implant fixation or other examined histomorphological patterns was not proved. A strong density of mast cells with a minimum presence of dendritic cells was observed in the control patient group. DISCUSSION The differences in density of S100 positive dendritic cells between the control and examined group of patients can be caused by the activation of dendritic cells by exogenous or endogenous pathways of immune processes going on after the implantation of endoprosthesis. The statistically significant interrelation of mast cells, polypously formed pseudosynovium and cement wear particles can be explained at least in part as a tissue reaction induced by cement particles. CONCLUSIONS We proved the presence of two immunologically significant cell populations in periprosthetic tissues. The said findings indicate a conclusion of significant functional participation of mast and dendritic cells in pathogenesis of aseptic loosening and periprosthetic osteolysis. Nevertheless, this will have to be proved in another way and with the use of another method. Key words:dendritic cells, mast cells, aseptic loosening, total joint replacement, immune reaction, adverse reaction.


Assuntos
Células Dendríticas/imunologia , Prótese de Quadril/microbiologia , Prótese do Joelho/microbiologia , Mastócitos/imunologia , Falha de Prótese/efeitos adversos , Antígenos CD1/imunologia , Células Dendríticas/ultraestrutura , Articulação do Quadril/microbiologia , Articulação do Quadril/patologia , Articulação do Quadril/cirurgia , Prótese de Quadril/efeitos adversos , Humanos , Articulação do Joelho/microbiologia , Articulação do Joelho/patologia , Articulação do Joelho/cirurgia , Prótese do Joelho/efeitos adversos , Mastócitos/ultraestrutura , Microscopia/instrumentação , Proteínas Proto-Oncogênicas c-kit/imunologia , Reoperação/métodos , Proteínas S100/imunologia , Triptases/imunologia
9.
Cancer Med ; 7(12): 5920-5927, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30358164

RESUMO

In this study, the immunophenotype was retrospectively analyzed in 131 patients who received initial treatment for plasma cell myeloma (PCM) and the relationships of CD81 and CD117 with the clinicopathologic characteristics and prognosis were further evaluated. The Kaplan and Meier method and Cox regression survival analysis model were used to determine whether CD117 and CD81 were factors affecting the overall survival (OS) and progression-free survival (PFS) of PCM patients. CD117 and CD81 positivity was demonstrated in 35.88% and 40.46% of the 131 patients, respectively. Kaplan-Meier analysis showed that CD117 and CD81 were potential predictors of a patient's prognosis. Specifically, CD117(+) patients had longer PFS (P = 0.033) and OS (P = 0.002), while CD81(+) patients had shorter PFS (P = 0.001) and OS (P = 0.002). CD117(+) and CD81(-) patients had the longest PFS [P = 0.0183 compared to the CD117(-)CD81(-)/CD117(+)CD81(+) group; P = 0.0007 compared to the CD117(-)CD81(+) group] and the longest OS [P = 0.0331 compared to the CD117(-)CD81(-)/CD117(+)CD81(+) group; P = 0.0005 compared to the CD117(-)CD81(+) group]. Our results show that CD81 is an independent factor affecting the OS and PFS of PCM patients, and CD117 is an independent factor affecting the OS of PCM patients. CD117-positive and CD81-negative patients with PCM have a better prognosis.


Assuntos
Mieloma Múltiplo/imunologia , Proteínas Proto-Oncogênicas c-kit/imunologia , Tetraspanina 28/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fenótipo , Prognóstico , Análise de Sobrevida
10.
Microsc Res Tech ; 81(11): 1268-1274, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30351479

RESUMO

Telocyte (TC) is an interesting unique interstitial cell demonstrated in many human and animal tissues and organs. This study verified, for the first time, the pattern of TC distribution in the testicular tissue of New Zealand White rabbits using histological, immunohistochemical, and electron microscopic tools. Rabbit testicular tissue samples were obtained from three pairs of adult healthy New Zealand white rabbit by surgical procedures. The testicular tissues were stained with hematoxyline-eosin, Crossmon's trichrome and Periodic acid Schiff. The immunohistochemistry was performed using three different antibodies CD34, CD117, and vimentin. The testes were examined by scanning and transmission electron microscopy. Histologically, TCs formed a sheath surrounding the seminiferous tubules. Other TCs were located in the interstitial tissue of the rabbit testis. Immunohistochemically, TCs reacted strongly with CD34, CD117, and vimentin. Scanning electron microscopic findings clearly elucidated the spreading pattern of TCs and their cytoplasmic processes with the interstitial tissue including blood vessels. Both homocellular and heterocellular junctions were demonstrated by transmission electron microscope. On the basis of TCs distribution and connections, the before mentioned data suggested that, TCs may play a potential role in maintaining the testicular construction and regulation. A future work is needed to clarify the actual role played by TCs in monitoring testicular fertility. RESEARCH HIGHLIGHTS: Telocyte (TC) is a unique cell demonstrated in human and animal tissues. TCs formed a sheath surrounding the seminiferous tubules in rabbits and may be located in interstitial tissue. Immunohistochemically, TCs reacted strongly with CD34 and CD117.


Assuntos
Células do Tecido Conjuntivo/ultraestrutura , Tecido Conjuntivo/anatomia & histologia , Telócitos/ultraestrutura , Testículo/anatomia & histologia , Testículo/citologia , Animais , Anticorpos/imunologia , Antígenos CD34/imunologia , Células do Tecido Conjuntivo/fisiologia , Imunofluorescência , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Proteínas Proto-Oncogênicas c-kit/imunologia , Coelhos , Telócitos/fisiologia , Vimentina/imunologia
11.
Am J Reprod Immunol ; 80(4): e13002, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29917288

RESUMO

Mast cells (MCs) are most known because of their prominent role in allergy. In the last years, however, it has become clear that they have pleiotropic functions and a high adaptability to the milieu. This review focuses on negative and positive roles of MCs within the body with particular emphasis on their participation in reproductive processes.


Assuntos
Hipersensibilidade/imunologia , Imunidade Inata/imunologia , Mastócitos/imunologia , Microambiente Celular/imunologia , Feminino , Humanos , Inflamação/imunologia , Gravidez , Proteínas Proto-Oncogênicas c-kit/imunologia , Receptores de IgE/imunologia
12.
J Agric Food Chem ; 66(22): 5581-5592, 2018 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-29763312

RESUMO

Deep-sea-derived butyrolactone I (BTL-I), which was identified as a type of butanolide, was isolated from Aspergillus sp. Ovalbumin (OVA)-induced BALB/c anaphylaxis was established to explore the antifood allergic activity of BTL-I. As a result, BTL-I was able to alleviate OVA-induced allergy symptoms, reduce the levels of histamine and mouse mast cell proteinases, inhibit OVA-specific IgE, and decrease the population of mast cells in the spleen and mesenteric lymph nodes. BTL-I also significantly suppressed mast-dependent passive cutaneous anaphylaxis. Additionally, the maturation of bone marrow-derived mast cells (BMMCs) declined as BTL-I caused down-regulation of c-KIT receptors. Furthermore, molecular docking analyses revealed that BTL-I interacted with the inhibitory receptor, FcγRIIB. In conclusion, the reduction of mast cell function by deep-sea-derived BTL-I as well as its interactions with the inhibitory receptor, FcγRIIB, may contribute to BTL-I-related protection against food anaphylaxis.


Assuntos
4-Butirolactona/análogos & derivados , Anafilaxia/tratamento farmacológico , Aspergillus/química , Hipersensibilidade Alimentar/tratamento farmacológico , Mastócitos/imunologia , 4-Butirolactona/administração & dosagem , Anafilaxia/genética , Anafilaxia/imunologia , Animais , Aspergillus/genética , Aspergillus/isolamento & purificação , Células Cultivadas , Modelos Animais de Doenças , Feminino , Hipersensibilidade Alimentar/genética , Hipersensibilidade Alimentar/imunologia , Histamina/imunologia , Humanos , Imunoglobulina E/imunologia , Mastócitos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/efeitos adversos , Proteínas Proto-Oncogênicas c-kit/genética , Proteínas Proto-Oncogênicas c-kit/imunologia , Água do Mar/microbiologia
13.
Clin Cancer Res ; 24(17): 4297-4308, 2018 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-29764854

RESUMO

Purpose: c-KIT overexpression is well recognized in cancers such as gastrointestinal stromal tumors (GIST), small cell lung cancer (SCLC), melanoma, non-small cell lung cancer (NSCLC), and acute myelogenous leukemia (AML). Treatment with the small-molecule inhibitors imatinib, sunitinib, and regorafenib resulted in resistance (c-KIT mutant tumors) or limited activity (c-KIT wild-type tumors). We selected an anti-c-KIT ADC approach to evaluate the anticancer activity in multiple disease models.Experimental Design: A humanized anti-c-KIT antibody LMJ729 was conjugated to the microtubule destabilizing maytansinoid, DM1, via a noncleavable linker (SMCC). The activity of the resulting ADC, LOP628, was evaluated in vitro against GIST, SCLC, and AML models and in vivo against GIST and SCLC models.Results: LOP628 exhibited potent antiproliferative activity on c-KIT-positive cell lines, whereas LMJ729 displayed little to no effect. At exposures predicted to be clinically achievable, LOP628 demonstrated single administration regressions or stasis in GIST and SCLC xenograft models in mice. LOP628 also displayed superior efficacy in an imatinib-resistant GIST model. Further, LOP628 was well tolerated in monkeys with an adequate therapeutic index several fold above efficacious exposures. Safety findings were consistent with the pharmacodynamic effect of neutropenia due to c-KIT-directed targeting. Additional toxicities were considered off-target and were consistent with DM1, such as effects in the liver and hematopoietic/lymphatic system.Conclusions: The preclinical findings suggest that the c-KIT-directed ADC may be a promising therapeutic for the treatment of mutant and wild-type c-KIT-positive cancers and supported the clinical evaluation of LOP628 in GIST, AML, and SCLC patients. Clin Cancer Res; 24(17); 4297-308. ©2018 AACR.


Assuntos
Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Imunoconjugados/farmacologia , Neoplasias/tratamento farmacológico , Proteínas Proto-Oncogênicas c-kit/genética , Animais , Anticorpos Anti-Idiotípicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/imunologia , Xenoenxertos , Humanos , Mesilato de Imatinib/farmacologia , Imunoconjugados/imunologia , Camundongos , Mutação , Neoplasias/classificação , Neoplasias/imunologia , Neoplasias/patologia , Inibidores de Proteínas Quinases/farmacologia , Proteínas Proto-Oncogênicas c-kit/imunologia
14.
Mol Ther ; 26(5): 1181-1197, 2018 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-29622475

RESUMO

We report a novel approach to bone marrow (BM) conditioning using c-kit-targeted chimeric antigen receptor T (c-kit CAR-T) cells in mice. Previous reports using anti-c-kit or anti-CD45 antibody linked to a toxin such as saporin have been promising. We developed a distinctly different approach using c-kit CAR-T cells. Initial studies demonstrated in vitro killing of hematopoietic stem cells by c-kit CAR-T cells but poor expansion in vivo and poor migration of CAR-T cells into BM. Pre-treatment of recipient mice with low-dose cyclophosphamide (125 mg/kg) together with CXCR4 transduction in the CAR-T cells enhanced trafficking to and expansion in BM (<1%-13.1%). This resulted in significant depletion of the BM c-kit+ population (9.0%-0.1%). Because congenic Thy1.1 CAR-T cells were used in the Thy1.2-recipient mice, anti-Thy1.1 antibody could be used to deplete CAR-T cells in vivo before donor BM transplant. This achieved 20%-40% multilineage engraftment. We applied this conditioning to achieve an average of 28% correction of chronic granulomatous disease mice by wild-type BM transplant. Our findings provide a proof of concept that c-kit CAR-T cells can achieve effective BM conditioning without chemo-/radiotherapy. Our work also demonstrates that co-expression of a trafficking receptor can enhance targeting of CAR-T cells to a designated tissue.


Assuntos
Células-Tronco Hematopoéticas/metabolismo , Proteínas Proto-Oncogênicas c-kit/imunologia , Receptores de Antígenos de Linfócitos T/metabolismo , Receptores de Antígenos Quiméricos/metabolismo , Linfócitos T/imunologia , Linfócitos T/metabolismo , Condicionamento Pré-Transplante , Animais , Biomarcadores , Células da Medula Óssea/metabolismo , Linhagem Celular , Técnicas de Cocultura , Citotoxicidade Imunológica , Citometria de Fluxo , Ordem dos Genes , Vetores Genéticos/genética , Imunofenotipagem , Camundongos , Camundongos Transgênicos , Condicionamento Pré-Transplante/métodos
15.
Biol Blood Marrow Transplant ; 24(8): 1554-1562, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29684562

RESUMO

Allogeneic hematopoietic stem cell transplantation (HSCT) can be curative for patients with sickle cell disease (SCD). However, morbidity associated with myeloablative conditioning and graft-versus-host disease has limited its utility. To this end, autologous HSCT for SCD using lentiviral gene-modified bone marrow (BM) or peripheral blood stem cells has been undertaken, although toxicities of fully ablative conditioning with busulfan and incomplete engraftment have been encountered. Treosulfan, a busulfan analog with a low extramedullary toxicity profile, has been used successfully as part of a myeloablative conditioning regimen in the allogeneic setting in SCD. To further minimize toxicity of conditioning, noncytotoxic monoclonal antibodies that clear stem cells from the marrow niche, such as anti-c-Kit (ACK2), have been considered. Using a murine model of SCD, we sought to determine whether nonmyeloablative conditioning followed by transplantation with syngeneic BM cells could ameliorate the disease phenotype. Treosulfan and ACK2, in a dose-dependent manner, decreased BM cellularity and induced cytopenia in SCD mice. Conditioning with treosulfan alone at nonmyeloablative dosing (3.6 g/kg), followed by transplantation with syngeneic BM donor cells, permitted long-term mixed-donor chimerism. Level of chimerism correlated with improvement in hematologic parameters, normalization of urine osmolality, and improvement in liver and spleen pathology. Addition of ACK2 to treosulfan conditioning did not enhance engraftment. Our data suggests that pretransplant conditioning with treosulfan alone may allow sufficient erythroid engraftment to reverse manifestations of SCD, with clinical application as a preparative regimen in SCD patients undergoing gene-modified autologous HSCT.


Assuntos
Anemia Falciforme/terapia , Transplante de Medula Óssea/métodos , Bussulfano/análogos & derivados , Condicionamento Pré-Transplante/métodos , Animais , Anticorpos/uso terapêutico , Antineoplásicos Alquilantes/uso terapêutico , Bussulfano/uso terapêutico , Modelos Animais de Doenças , Sobrevivência de Enxerto , Camundongos , Proteínas Proto-Oncogênicas c-kit/imunologia , Resultado do Tratamento
17.
Vet Comp Oncol ; 16(1): E176-E184, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29152836

RESUMO

Non-adherent, 3-dimensional sphere formation is used as an in vitro surrogate to evaluate cellular potential for tumour initiation and self-renewal. To determine if a shared molecular program underlies the capacity for sphere formation by cells originating from diverse tumour types, we characterized molecular and functional properties of 10 independent cell lines derived from 3 ontogenetically distinct dog cancers: hemangiosarcoma, osteosarcoma and glial brain tumours. Genome-wide gene expression profiling identified tumour-of-origin-dependent patterns of adjustment to sphere formation in a uniform culture condition. However, expression of the stem/progenitor markers CD34 and CD117, resistance to cytotoxic drugs and dye efflux (side population assays) showed no association with these gene expression profiles. Instead, primary sphere-forming capacity was inversely correlated with the ability to reform secondary spheres, regardless of tumour ontogeny. Primary sphere formation seemed to be proportional to the number of pre-existing cells with sphere-forming capacity in the cell lines. Cell lines where secondary sphere formation was more proficient than primary sphere formation showed enrichment of genes involved in fatty acid synthesis and immunosuppressive cytokines. In contrast, cell lines where secondary sphere formation was approximately equivalent to or less proficient than primary sphere formation showed upregulation of CD40 and enrichment of genes involved in fatty acid oxidation. Our data suggest that in vitro sphere formation is associated with upregulation of gene clusters involved in metabolic and immunosuppressive functions, which might be necessary for self-renewal and for tumour initiation and/or tumour propagation in vivo.


Assuntos
Doenças do Cão/metabolismo , Ácidos Graxos/metabolismo , Tolerância Imunológica , Neoplasias/veterinária , Animais , Antígenos CD34/imunologia , Antígenos CD40/imunologia , Linhagem Celular Tumoral , Doenças do Cão/imunologia , Cães , Técnicas In Vitro , Neoplasias/imunologia , Neoplasias/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Proteínas Proto-Oncogênicas c-kit/imunologia , RNA Neoplásico/genética , Transcriptoma/imunologia
18.
J Allergy Clin Immunol ; 141(1): 180-188.e3, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28629749

RESUMO

BACKGROUND: Clonal mast cell disorders are known to occur in a subset of patients with systemic reactions to Hymenoptera stings. This observation has prompted the question of whether clonal mast cell disorders also occur in patients with idiopathic anaphylaxis (IA). OBJECTIVE: We sought to determine the prevalence of clonal mast cell disorders among patients with IA, criteria to identify those patients who require a bone marrow biopsy, and whether the pathogenesis of IA involves a hyperresponsive mast cell compartment. METHODS: We prospectively enrolled patients with IA (≥3 episodes/y) who then underwent a medical evaluation that included a serum tryptase determination, allele-specific quantitative PCR (ASqPCR) for the KIT D816V mutation, and a bone marrow examination. Mast cells were cultured from peripheral blood CD34+ cells and examined for releasability after FcεRI aggregation. RESULTS: Clonal mast cell disease was diagnosed in 14% of patients referred with IA. ASqPCR for the KIT D816V mutation was a useful adjunct in helping identify those with systemic mastocytosis but not monoclonal mast cell activation syndrome. A modified overall clonal prediction model was developed by using clinical findings, a serum tryptase determination, and ASqPCR. There was no evidence of a hyperresponsive mast cell phenotype in patients with IA. CONCLUSION: Patients with clonal mast cell disease can present as having IA. Distinct clinical and laboratory features can be used to select those patients more likely to have an underlying clonal mast cell disorder (monoclonal mast cell activation syndrome or systemic mastocytosis) and thus candidates for a bone marrow biopsy.


Assuntos
Anafilaxia/genética , Anafilaxia/imunologia , Mastócitos/imunologia , Mastocitose Sistêmica/genética , Mastocitose Sistêmica/imunologia , Mutação de Sentido Incorreto , Proteínas Proto-Oncogênicas c-kit , Adolescente , Adulto , Idoso , Substituição de Aminoácidos , Anafilaxia/patologia , Feminino , Humanos , Masculino , Mastócitos/patologia , Mastocitose Sistêmica/patologia , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas c-kit/genética , Proteínas Proto-Oncogênicas c-kit/imunologia
19.
Nat Commun ; 8(1): 1911, 2017 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-29203769

RESUMO

Murine γδ T cells include subsets that are programmed for distinct effector functions during their development in the thymus. Under pathological conditions, different γδ T cell subsets can be protective or can exacerbate a disease. Here we show that CD117, CD200 and CD371, together with other markers, identify seven developmental stages of γδ T cells. These seven stages can be divided into three distinct developmental pathways that are enriched for different TCRδ repertoires and exhibit characteristic expression patterns associated with adaptive (γδTn), IFN-γ-producing (γδT1) and IFN-γ/IL-4-co-producing γδ T cells (γδNKT). Developmental progression towards both IFN-γ-producing subsets can be induced by TCR signalling, and each pathway results in thymic emigration at a different stage. Finally, we show that γδT1 cells are the predominating IFN-γ-producing subset developing in the adult thymus. Thus, this study maps out three distinct development pathways that result in the programming of γδTn, γδT1 and γδNKT cells.


Assuntos
Linfopoese/imunologia , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T/imunologia , Timócitos/imunologia , Animais , Antígenos CD/imunologia , Diferenciação Celular , Interferon gama/imunologia , Interleucina-4/imunologia , Camundongos , Células T Matadoras Naturais/citologia , Células T Matadoras Naturais/imunologia , Proteínas Proto-Oncogênicas c-kit/imunologia , Transdução de Sinais , Subpopulações de Linfócitos T/citologia , Linfócitos T/citologia , Timócitos/citologia , Timo/citologia
20.
Infect Immun ; 85(11)2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28784931

RESUMO

Alcohol abuse impairs immune defense. To study the effect of chronic-plus-binge alcohol exposure on the granulopoietic response, acute alcohol intoxication (intraperitoneal injection of 5 g alcohol/kg body weight) was introduced to mice chronically fed on the Lieber-DeCarli low-fat liquid alcohol diet for 5 weeks. Bacteremia was induced by intravenous injection of Escherichia coli Bacteremia caused a remarkable increase in marrow lin- c-kit+ Sca-1+ cells. Activation of cell proliferation supported the increase in marrow lin- c-kit+ Sca-1+ cells. Alcohol administration inhibited this activation of lin- c-kit+ Sca-1+ cells. The bone marrow of pair-fed control mice receiving intraperitoneal saline stored a large number of mature granulocytes expressing a high level of Gr1 (Gr1hi cells). The proportion of Gr1hi cells and the total number of Gr1+ cells were markedly reduced in the bone marrow, along with an increase in the ratio of Gr1+ granulocytes in peripheral white blood cells following bacteremia. E. coli infection stimulated proliferation of granulopoietic precursor cells, resulting in a marked increase in the ratio of immature Gr1lo cells in the bone marrow. Alcohol administration itself triggered marrow release of Gr1+ cells, resulting in reduction of the marrow granulocyte reserve with an elevation of granulocytes in the circulation. Alcohol also impaired activation of granulopoietic precursor proliferation following bacteremia. Alcohol disrupted lipopolysaccharide (LPS)-TLR4-ERK1/2-cyclin D1 signaling and inhibited upregulation of Sca-1 and C/EBPß expression by lineage-negative marrow cells in response to bacteremia. These results indicate that chronic-plus-binge alcohol exposure inhibits the granulopoietic response by disrupting key cell signaling for hematopoietic precursor cell activation and commitment to granulocyte lineage development.


Assuntos
Bacteriemia/imunologia , Bebedeira/imunologia , Infecções por Escherichia coli/imunologia , Etanol/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Hematopoese/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Animais , Antígenos Ly/genética , Antígenos Ly/imunologia , Bacteriemia/genética , Bacteriemia/patologia , Bebedeira/genética , Bebedeira/patologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/imunologia , Células da Medula Óssea/patologia , Proteína beta Intensificadora de Ligação a CCAAT/genética , Proteína beta Intensificadora de Ligação a CCAAT/imunologia , Ciclina D1/genética , Ciclina D1/imunologia , Modelos Animais de Doenças , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/imunologia , Infecções por Escherichia coli/genética , Infecções por Escherichia coli/patologia , Regulação da Expressão Gênica/imunologia , Granulócitos/efeitos dos fármacos , Granulócitos/imunologia , Granulócitos/patologia , Hematopoese/genética , Hematopoese/imunologia , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/imunologia , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/imunologia , Nucleotidiltransferases/deficiência , Nucleotidiltransferases/genética , Nucleotidiltransferases/imunologia , Proteínas Proto-Oncogênicas c-kit/genética , Proteínas Proto-Oncogênicas c-kit/imunologia , Transdução de Sinais/imunologia , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia
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