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1.
Can J Physiol Pharmacol ; 101(6): 316-326, 2023 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-36867857

RESUMO

Introduction: This study evaluated depression, monocyte phenotype, and immune function in physically active cannabis users. Methods: Participants (N = 23) were classified as either cannabis users (CU, n = 11) or non-users (NU, n = 12). White blood cells isolated from blood were analyzed for co-expression of cluster of differentiation 14 and 16 using flow cytometry. Lipopolysaccharide (LPS) was cultured with whole blood and assessed for interleukin-6 and tumor necrosis factor-α (TNF-α) release. Results: The percentage of white blood cells classified as monocytes was not different between groups; however, CU had a significantly greater percentage of monocytes classified as intermediate (p = 0.02). When standardized per milliliter of blood, CU had significantly greater numbers of total monocytes (p = 0.01), classical monocytes (p = 0.02), and intermediate monocytes (p = 0.01). Intermediate monocytes per milliliter of blood were positively correlated to the number of times CU used cannabis per day (r = 0.864, p < 0.01) and Beck Depression Inventory-II (BDI-II) score (r = 0.475, p = 0.03), which was significantly greater in CU (5.1 ± 4.8) compared with NU (0.8 ± 1.0; p < 0.01). CU released significantly less TNF-α per monocyte in response to LPS. Conclusions: CU had altered monocyte phenotypes and functions compared with NU. Elevations in intermediate monocytes were positively correlated with measures of cannabis use and BDI-II score.


Assuntos
Cannabis , Monócitos , Cannabis/efeitos adversos , Fator de Necrose Tumoral alfa , Depressão/complicações , Lipopolissacarídeos/farmacologia , Fenótipo , Imunidade
2.
Int J Mol Sci ; 23(18)2022 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-36142210

RESUMO

Immunotherapies relying on type 1 immunity have shown robust clinical responses in some cancers yet remain relatively ineffective in solid breast tumors. Polarization toward type 2 immunity and expansion of myeloid-derived suppressor cells (MDSC) confer resistance to therapy, though it remains unclear whether polarization toward type 3 immunity occurs or has a similar effect. Therefore, we investigated the involvement of type 3 Th17 and Th22 cells and their association with expanding MDSC populations in the 4T1 mouse mammary carcinoma model. Th17 and Th22 were detected in the earliest measurable mass at d 14 and remained present until the final sampling on d 28. In peripheral organs, Th17 populations were significantly higher than the non-tumor bearing control and peaked early at d 7, before a palpable tumor had formed. Peripheral Th22 proportions were also significantly increased, though at later times when tumors were established. To further address the mechanism underlying type 3 immune cell and MDSC recruitment, we used CRISPR-Cas9 to knock out 4T1 tumor production of interleukin-6 (4T1-IL-6-KO), which functions in myelopoiesis, MDSC recruitment, and Th maturation. While 4T1-IL-6-KO tumor growth was similar to the control, the reduced IL-6 significantly expanded the total CD4+ Th population and Th17 in tumors, while Th22 and MDSC were reduced in all tissues; this suggests that clinical IL-6 depletion combined with immunotherapy could improve outcomes. In sum, 4T1 mammary carcinomas secrete IL-6 and other factors, to polarize and reshape Th populations and expand distinct Th17 and Th22 populations, which may facilitate tumor growth and confer immunotherapy resistance.


Assuntos
Carcinoma , Células Supressoras Mieloides , Animais , Carcinoma/patologia , Imunoterapia , Interleucina-6 , Interleucinas , Camundongos , Dinâmica Populacional , Células Th17
3.
Phytomedicine ; 105: 154343, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35901597

RESUMO

BACKGROUND: Our previous research revealed a novel function of berberine (BBR), a clinically relevant plant-derived alkaloid, as a suppressor of follicular T helper (Tfh) cell proliferation in secondary lymphoid organs of BBR-treated mice that underwent immunization for collagen-induced arthritis (CIA) in DBA1/J mice. Due to the importance of Tfh cell and B cell interactions in the generation of T cell-dependent humoral responses, the suppression of Tfh cell activity may have implications for the general safety of BBR as a prophylactic dietary supplement, and its potential use in antibody-driven autoimmune and hypersensitivity disorders. PURPOSE: This research aims to characterize BBR's impact on the activation, differentiation, and proliferation of Tfh cells by examining the expression of key extracellular signaling molecules, as well as the activity of intracellular signaling molecules involved in the Ca2+-calcineurin-NFAT pathway and STAT3 phosphorylation, following activation. STUDY DESIGN: In vitro experimental study using primary tissues. METHODS: To explore the direct effects of BBR on the proliferation and differentiation of Tfh cells, isolated naïve CD4+ T cells (>95% pure) were activated and differentiated into pre- Tfh cells in the presence or absence of BBR. The resulting Tfh cell populations and the expression of the key extracellular molecules CXCR5, ICOS, and PD-1 were measured. In addition, we examined the impact of BBR treatment on the activity of key intracellular signaling molecules involved in Tfh cell activation and differentiation following TCR ligation and/or CD28 signaling (p-ZAP-70, p-Lck, p-PLCγ1, NFATc1 and intracellular calcium, Ca2+, concentrations), as well as IL-6 signaling (p-STAT3). RESULTS: Treatment with BBR significantly reduced the expression of both CXCR5 (p < 0.01) and ICOS (p < 0.005), but not PD-1, and reduced the percentage of Tfh cells within the total CD4+ T cell population. BBR treatment also led to a reduction in intracellular Ca2+ flux, activation of p-STAT3, and IL-21 production. CONCLUSION: Our observations provide insight into the mechanism of BBR-mediated Tfh cell suppression and suggest that BBR treatment can directly inhibit Tfh cell activity, perhaps through interfering with cytokine receptor or downstream signaling.


Assuntos
Berberina , Linfócitos T Auxiliares-Indutores , Animais , Citocinas , Terapia de Imunossupressão , Camundongos , Receptores CXCR5
4.
Data Brief ; 41: 108009, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35252506
5.
Med Sci Sports Exerc ; 53(10): 2045-2053, 2021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34519716

RESUMO

PURPOSE: This study investigated the effects of 12 wk of postexercise kefir consumption in cancer survivors who have undergone chemotherapy and/or radiation therapy. METHODS: All participants were enrolled in a structured exercise training program and separated into kefir (KEF) or control (CON) treatment groups. KEF consumed 8 oz. of kefir after exercise sessions (3 d·wk-1) for 12 wk. Outcome measures included assessments for body size and composition, aerobic fitness and muscular strength, medical history, and psychological state at pre- and postintervention time points. Blood was collected and analyzed for C-reactive protein (CRP), interleukin 6 (IL-6), and lipopolysaccharide (LPS) concentrations, and LPS-stimulated whole blood IL-6 and tumor necrosis factor α production were obtained using enzyme-linked immunosorbent assays at both time points. Monocyte numbers and phenotype were obtained using flow cytometry. RESULTS: Participants (N = 24; 9 males and 15 females) were an average of 61 ± 9.9 yr old. Kefir consumption was associated with 6.3% (P = 0.034) improvements in lean body mass, as well as 51.4% (P = 0.046), 39.3% (P = 0.017), and 64.7% (P = 0.021) improvements in measures of depression, fatigue, and gastric distress, respectively. KEF also experienced a significant 35.4% (P = 0.01) reduction in circulating LPS along with an 18.0% increase (P < 0.001) in classical monocytes % and a 22.3% decrease (P = 0.04) in nonclassical monocytes %. There were no significant changes in any other variables. CONCLUSION: Twelve weeks of kefir consumption improved lean body mass, depression, fatigue, gastric distress, and a biomarker of gut dysbiosis. Kefir improved overall and classical monocyte numbers. Kefir should be considered as a component of a postexercise dietary regimen for cancer survivors.


Assuntos
Sobreviventes de Câncer , Terapia por Exercício , Kefir , Adulto , Idoso , Antineoplásicos/efeitos adversos , Biomarcadores/sangue , Contagem de Células Sanguíneas , Índice de Massa Corporal , Sobreviventes de Câncer/psicologia , Aptidão Cardiorrespiratória , Citocinas/sangue , Depressão/prevenção & controle , Fadiga/prevenção & controle , Feminino , Gastroenteropatias/etiologia , Gastroenteropatias/prevenção & controle , Humanos , Inflamação/prevenção & controle , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , Força Muscular , Qualidade de Vida , Radioterapia/efeitos adversos
6.
Int J Mol Sci ; 22(6)2021 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-33806895

RESUMO

Skeletal muscle regeneration is highly dependent on the inflammatory response. A wide variety of innate and adaptive immune cells orchestrate the complex process of muscle repair. This review provides information about the various types of immune cells and biomolecules that have been shown to mediate muscle regeneration following injury and degenerative diseases. Recently developed cell and drug-based immunomodulatory strategies are highlighted. An improved understanding of the immune response to injured and diseased skeletal muscle will be essential for the development of therapeutic strategies.


Assuntos
Imunidade Adaptativa , Imunidade Inata , Músculo Esquelético/fisiologia , Regeneração/imunologia , Fatores Etários , Animais , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Suscetibilidade a Doenças , Humanos , Imunomodulação , Leucócitos/imunologia , Leucócitos/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Desenvolvimento Muscular/genética , Desenvolvimento Muscular/imunologia , Linfócitos T/imunologia , Linfócitos T/metabolismo
7.
Int J Mol Sci ; 22(7)2021 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-33805383

RESUMO

There is evidence that berberine (BBR), a clinically relevant plant compound, ameliorates clinically apparent collagen-induced arthritis (CIA) in vivo. However, to date, there are no studies involving the use of BBR which explore its prophylactic potential in this model of rheumatoid arthritis (RA). The aim of this study was to determine if prophylactic BBR use during the preclinical phase of collagen-induced arthritis would delay arthritic symptom onset, and to characterize the cellular mechanism underlying such an effect. DBA/1J mice were injected with an emulsion of bovine type II collagen (CII) and complete Freund's adjuvant (day 0) and a booster injection of CII in incomplete Freund's adjuvant (day 18) to induce arthritis. Mice were then given i.p. injections of 1 mg/kg/day of BBR or PBS (vehicle with 0.01% DMSO) from days 0 to 28, were left untreated (CIA control), or were in a non-arthritic control group (n = 15 per group). Incidence of arthritis in BBR-treated mice was 50%, compared to 90% in both the CIA and PBS controls. Populations of B and T cells from the spleens and draining lymph nodes of mice were examined on day 14 (n = 5 per group) and day 28 (n = 10 per group). BBR-treated mice had significantly reduced populations of CD4+Th and CD4+CXCR5+ Tfh cells, and an increased proportion of Foxp3+ Treg at days 14 and 28, as well as reduced expression of co-stimulatory molecules CD28 and CD154 at both endpoints. The effect seen on T cell populations and co-stimulatory molecule expression in BBR-treated mice was not mirrored in CD19+ B cells. Additionally, BBR-treated mice experienced reduced anti-CII IgG2a and anti-CII total IgG serum concentrations. These results indicate a potential role for BBR as a prophylactic supplement for RA, and that its effect may be mediated specifically through T cell suppression. However, the cellular effector involved raises concern for BBR prophylactic use in the context of vaccine efficacy and other primary adaptive immune responses.


Assuntos
Artrite Experimental/prevenção & controle , Berberina/farmacologia , Linfócitos T/efeitos dos fármacos , Animais , Artrite Experimental/induzido quimicamente , Artrite Experimental/imunologia , Artrite Reumatoide/induzido quimicamente , Artrite Reumatoide/imunologia , Artrite Reumatoide/prevenção & controle , Linfócitos B , Berberina/uso terapêutico , Colágeno Tipo II/toxicidade , Imunoglobulina G/sangue , Camundongos , Camundongos Endogâmicos DBA , Substâncias Protetoras/farmacologia , Substâncias Protetoras/uso terapêutico , Linfócitos T/imunologia
8.
PLoS One ; 15(6): e0234548, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32542046

RESUMO

BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are potent suppressors of immune function and may play a key role in the development and progression of metastatic cancers. Aerobic exercise has been shown to have anticancer effects, yet the mechanisms behind this protection are largely unknown. Therefore, we examined the effects of physical activity on MDSC accumulation and function. METHODS: Female BALB/c mice were assigned to one of two primary groups: sedentary tumor (SED+TUM) or wheel run tumor (WR+TUM). After 6 weeks of voluntary wheel running, all animals were randomly subdivided into 4 different timepoint groups; 16, 20, 24, and 28 days post-tumor injection. All mice were inoculated with 4T1 mammary carcinoma cells in the mammary fat pad and WR groups continued to run for the specified time post-injection. Spleen, blood, and tumor samples were analyzed using flow cytometry to assess proportions of MDSCs. RESULTS: Cells expressing MDSC biomarkers were detected in the spleen, blood, and tumor beginning at d16. However, since there was no evidence of immunosuppressive function until d28, we refer to them as immature myeloid cells (IMCs). Compared to SED+TUM, levels of IMCs in the spleen were significantly lower (p < 0.05) in WR+TUM at day 16 (33.0 ± 5.2%; 23.1 ± 10.2% of total cells, respectively) and day 20 (33.9 ± 8.1%; 24.3 ± 5.1% of total cells, respectively). Additionally, there were fewer circulating IMCs in WR+TUM at day 16 and MDSC levels were significantly lower (p < 0.05) in the tumor at day 28 in WR+TUM. Additionally, a non-significant 62% and 26% reduction in metastatic lung nodules was observed at days 24 and 28, respectively. At day 28, MDSCs harvested from SED+TUM significantly suppressed CD3+CD4+ T cell proliferation (3.2 ± 1.3 proliferation index) while proliferation in WR+TUM MDSC co-cultures (5.1 ± 1.7 proliferation index) was not different from controls. CONCLUSIONS: These findings suggest that physical activity may delay the accumulation of immunosuppressive MDSCs providing a broader window of opportunity for interventions with immunotherapies.


Assuntos
Terapia de Imunossupressão , Neoplasias Mamárias Experimentais/metabolismo , Células Supressoras Mieloides/metabolismo , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Técnicas de Cocultura , Feminino , Imunossupressores/farmacologia , Ativação Linfocitária/genética , Ativação Linfocitária/fisiologia , Neoplasias Mamárias Experimentais/genética , Camundongos , Atividade Motora/genética , Atividade Motora/fisiologia , Células Mieloides/metabolismo , Células Mieloides/patologia , Células Supressoras Mieloides/patologia , Células Supressoras Mieloides/fisiologia
9.
Int J Mol Sci ; 21(4)2020 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-32098318

RESUMO

Mast cells are often regarded through the lens of IgE-dependent reactions as a cell specialized only for anti-parasitic and type I hypersensitive responses. However, recently many researchers have begun to appreciate the expansive repertoire of stimuli that mast cells can respond to. After the characterization of the interleukin (IL)-33/suppression of tumorigenicity 2 (ST2) axis of mast cell activation-a pathway that is independent of the adaptive immune system-researchers are revisiting other stimuli to induce mast cell activation and/or subsequent degranulation independent of IgE. This discovery also underscores that mast cells act as important mediators in maintaining body wide homeostasis, especially through barrier defense, and can thus be the source of disease as well. Particularly in the gut, inflammatory bowel diseases (Crohn's disease, ulcerative colitis, etc.) are characterized with enhanced mast cell activity in the context of autoimmune disease. Mast cells show phenotypic differences based on tissue residency, which could manifest as different receptor expression profiles, allowing for unique mast cell responses (both IgE and non-IgE mediated) across varying tissues as well. This variety in receptor expression suggests mast cells respond differently, such as in the gut where immunosuppressive IL-10 stimulates the development of food allergy or in the lungs where transforming growth factor-ß1 (TGF-ß1) can enhance mast cell IL-6 production. Such differences in receptor expression illustrate the truly diverse effector capabilities of mast cells, and careful consideration must be given toward the phenotype of mast cells observed in vitro. Given mast cells' ubiquitous tissue presence and their capability to respond to a broad spectrum of non-IgE stimuli, it is expected that mast cells may also contribute to the progression of autoimmune disorders and other disease states such as metastatic cancer through promoting chronic inflammation in the local tissue microenvironment and ultimately polarizing toward a unique Th17 immune response. Furthermore, these interconnected, atypical activation pathways may crosstalk with IgE-mediated signaling differently across disorders such as parasitism, food allergies, and autoimmune disorders of the gut. In this review, we summarize recent research into familiar and novel pathways of mast cells activation and draw connections to clinical human disease.


Assuntos
Colite Ulcerativa/imunologia , Doença de Crohn/imunologia , Imunoglobulina E/imunologia , Interleucina-6/imunologia , Mastócitos/imunologia , Animais , Colite Ulcerativa/metabolismo , Doença de Crohn/metabolismo , Hipersensibilidade Alimentar/imunologia , Hipersensibilidade Alimentar/metabolismo , Humanos , Imunoglobulina E/metabolismo , Proteína 1 Semelhante a Receptor de Interleucina-1/imunologia , Proteína 1 Semelhante a Receptor de Interleucina-1/metabolismo , Interleucina-33/imunologia , Interleucina-33/metabolismo , Interleucina-6/metabolismo , Mastócitos/metabolismo
10.
J Leukoc Biol ; 106(1): 95-103, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30924969

RESUMO

Gut immune system homeostasis involves diverse structural interactions among resident microbiota, the protective mucus layer, and a variety of cells (intestinal epithelial, lymphoid, and myeloid). Due to the substantial surface area in direct contact with an "external" environment and the diversity of xenobiotic, abiotic, and self-interactions coordinating to maintain gut homeostasis, there is enhanced potential for the generation of endogenous danger signals when this balance is lost. Here, we focus on the potential generation and reception of damage in the gut resulting from exposure to nanoparticles (NPs), common food and drug additives. Specifically, we describe recent evidence in the literature showing that certain NPs are potential generators of damage-associated molecular patterns, as well as potential immune-stimulating molecular patterns themselves.


Assuntos
Alarminas/fisiologia , Mucosa Intestinal/efeitos dos fármacos , Nanopartículas/toxicidade , Trifosfato de Adenosina/metabolismo , Administração Oral , Animais , DNA Mitocondrial/fisiologia , Homeostase , Humanos , Mucosa Intestinal/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/fisiologia , Nanopartículas/metabolismo , Estresse Oxidativo/efeitos dos fármacos
11.
PLoS One ; 13(11): e0207704, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30444930

RESUMO

INTRODUCTION: For immune cells transforming growth factor beta-1 (TGF-ß1) can enhance or repress effector functions. Here, we characterize the effects of TGF-ß1 on IgE-mediated and IL-33-mediated activation of primary murine mast cells derived from hematopoietic stem cells (bone marrow derived mast cells; BMMC). We also investigated potential interactions between TGF-ß1 and stem cell factor (SCF). We conclude TGF-ß1 plays a selectively stimulatory role for mast cell cultures in vitro. METHODS: BMMCs from C57BL/6 mice were differentiated with IL-3 and then treated with TGF-ß1. BMMCs were exposed to TGF-ß1, primed with IgE, activated with antigen, and then IL-6 and IL-13 cytokine release was quantified using ELISA. Additionally, the effects of TGF-ß1 on both IgE and IL-33-mediated short term activation were observed via flow cytometric analysis of both surface LAMP-1 expression and intracellular IL-6. Receptor colocalization was visualized using fluorescence confocal microscopy and individual receptor expression levels were also quantified. RESULTS: Resting IL-6 production increased with TGF-ß1 but significance was lost following BMMC activation via IgE receptor (FcεRI) crosslinking. This was similar to a comparison effect due to SCF treatment alone, which also enhanced resting levels of IL-6. TGF-ß1 treatment enhanced release of IL-13 only with FcεRI-IgE-mediated activation. TGF-ß1 suppressed mobilization of IL-6 with short-term BMMC activation when stimulated with IL-33. Lastly, colocalization patterns of the SCF receptor (CD117) and FcεRI with IgE crosslinking were unaffected by TGF-ß1 treatment, but individual expression levels for FcεRI, CD117, and TGFßRII were all reduced following either IgE activation or TGF-ß1 treatment; this reduction was partially recovered in BMMCs that were both activated by IgE and treated with TGF-ß1. DISCUSSION: These data reveal a novel positive effect of soluble TGF-ß1 on mast cell activation in vitro, suggesting mast cells may be activated through a non-canonical pathway by TGF-ß1. Understanding this interaction will provide insight into the potential role of mast cells in settings where TGF-ß1 is produced in an aberrant manner, such as in and around high grade tumors.


Assuntos
Imunoglobulina E/metabolismo , Interleucina-13/metabolismo , Interleucina-6/metabolismo , Mastócitos/citologia , Fator de Crescimento Transformador beta1/farmacologia , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Citometria de Fluxo , Interleucina-33/farmacologia , Proteínas de Membrana Lisossomal/metabolismo , Mastócitos/efeitos dos fármacos , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Confocal
12.
Cell Immunol ; 290(1): 80-8, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24950026

RESUMO

A Disintegrin and Metalloproteinase (ADAM)-10 plays critical roles in neuronal migration and distribution. Recently, ADAM10 deletion was shown to disrupt myelopoiesis. We found that inducible deletion of ADAM10 using Mx1-driven Cre recombinase for a period of three weeks resulted in mast cell hyperplasia in the skin, intestine and spleen. Mast cells express surface ADAM10 in vitro and in vivo, at high levels compared to other immune cells tested. ADAM10 is important for mast cell migration, since ADAM10-deficiency reduced c-Kit-mediated migration. As with some mast cell proteases, ADAM10 expression could be altered by the cytokine microenvironment, being inhibited by IL-10 or TGFß1, but not by several other T cell-derived cytokines. Collectively these data show that the ADAM10 protease is an important factor in mast cell migration and tissue distribution, and can be manipulated by environmental cues.


Assuntos
Proteínas ADAM/metabolismo , Secretases da Proteína Precursora do Amiloide/metabolismo , Movimento Celular/genética , Mastócitos/fisiologia , Proteínas de Membrana/metabolismo , Fator de Células-Tronco/metabolismo , Proteínas ADAM/biossíntese , Proteínas ADAM/genética , Proteína ADAM10 , Secretases da Proteína Precursora do Amiloide/biossíntese , Secretases da Proteína Precursora do Amiloide/genética , Animais , Proliferação de Células , Células Cultivadas , Hiperplasia/genética , Interleucina-10/farmacologia , Proteínas de Membrana/biossíntese , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Peritônio/citologia , Interferência de RNA , RNA Interferente Pequeno , Linfócitos T/imunologia , Fator de Crescimento Transformador beta/farmacologia
13.
J Leukoc Biol ; 95(4): 643-50, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24338630

RESUMO

Mast cells and MDSCs are increased by parasitic infection and tumor growth. We previously demonstrated that enhanced MDSC development in ADAM10 transgenic mice yielded resistance to Nb infection and that coculturing MDSCs and mast cells enhanced cytokine production. In the current work, we show that MDSC-mast cell coculture selectively enhances IgE-mediated cytokine secretion among mast cells, without increasing MDSC cytokine production. This effect was independent of cell contact and elicited by Ly6C(+) and Ly6C/G+ MDSC subsets. These interactions were functionally important. MDSC depletion with the FDA-approved drug gemcitabine exacerbated Nb or Trichinella spiralis infection and reduced mast cell-dependent AHR and lung inflammation. Adoptive transfer of MDSC worsened AHR in WT but not mast cell-deficient Wsh/Wsh mice. These data support the hypothesis that MDSCs enhance mast cell inflammatory responses and demonstrate that this interaction can be altered by an existing chemotherapeutic.


Assuntos
Imunoglobulina E/imunologia , Mastócitos/imunologia , Células Mieloides/fisiologia , Animais , Asma/imunologia , Células Cultivadas , Citocinas/biossíntese , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Nippostrongylus/imunologia , Trichinella spiralis/imunologia
14.
J Immunol ; 191(9): 4505-13, 2013 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-24068671

RESUMO

We previously demonstrated that TGF-ß1 suppresses IgE-mediated signaling in human and mouse mast cells in vitro, an effect that correlated with decreased expression of the high-affinity IgE receptor, FcεRI. The in vivo effects of TGF-ß1 and the means by which it suppresses mast cells have been less clear. This study shows that TGF-ß1 suppresses FcεRI and c-Kit expression in vivo. By examining changes in cytokine production concurrent with FcεRI expression, we found that TGF-ß1 suppresses TNF production independent of FcεRI levels. Rather, IgE-mediated signaling was altered. TGF-ß1 significantly reduced expression of Fyn and Stat5, proteins critical for cytokine induction. These changes may partly explain the effects of TGF-ß1, because Stat5B overexpression blocked TGF-mediated suppression of IgE-induced cytokine production. We also found that Stat5B is required for mast cell migration toward stem cell factor, and that TGF-ß1 reduced this migration. We found evidence that genetic background may alter TGF responses. TGF-ß1 greatly reduced mast cell numbers in Th1-prone C57BL/6, but not Th2-prone 129/Sv mice. Furthermore, TGF-ß1 did not suppress IgE-induced cytokine release and did increase c-Kit-mediated migration in 129/Sv mast cells. These data correlated with high basal Fyn and Stat5 expression in 129/Sv cells, which was not reduced by TGF-ß1 treatment. Finally, primary human mast cell populations also showed variable sensitivity to TGF-ß1-mediated changes in Stat5 and IgE-mediated IL-6 secretion. We propose that TGF-ß1 regulates mast cell homeostasis, and that this feedback suppression may be dependent on genetic context, predisposing some individuals to atopic disease.


Assuntos
Imunoglobulina E/imunologia , Mastócitos/metabolismo , Receptores de IgE/imunologia , Fator de Transcrição STAT5/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Animais , Movimento Celular/imunologia , Células Cultivadas , Citocinas/metabolismo , Humanos , Imunoglobulina E/metabolismo , Mastócitos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Proto-Oncogênicas c-fyn/metabolismo , Proteínas Proto-Oncogênicas c-kit/metabolismo , Interferência de RNA , RNA Interferente Pequeno , Receptores de IgE/biossíntese , Receptores de IgE/metabolismo , Fator de Transcrição STAT5/genética , Fator de Transcrição STAT5/imunologia , Transdução de Sinais/imunologia , Fator de Crescimento Transformador beta1/imunologia , Fatores de Necrose Tumoral/biossíntese
15.
Biomaterials ; 34(18): 4439-51, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23515178

RESUMO

In this study, we investigated the effect of fiber and pore size of an electrospun scaffold on the polarization of mouse bone marrow-derived macrophages (BMMΦs) towards regenerative (M2) or inflammatory (M1) phenotypes. BMMΦs were seeded on Polydioxanone (PDO) scaffolds electrospun from varying polymer concentrations (60, 100, and 140 mg/ml). Higher polymer concentrations yielded larger diameter fibers with larger pore sizes and porosity. BMMΦ cultured on these scaffolds showed a correlation between increasing fiber/pore size and increased expression of the M2 marker Arginase 1 (Arg1), along with decreased expression of the M1 marker inducible nitric oxide synthase (iNOS). Secretion of the angiogenic cytokines VEGF, TGF-ß1 and bFGF was higher among cultures employing larger fiber/pore size scaffolds (140 mg/ml). Using a 3D in vitro angiogenesis bead assay, we have demonstrated that the M2-like profile of BMMΦ induced by the 140 mg/ml is functional. Furthermore, our results show that the pore size of a scaffold is a more critical regulator of the BMMΦ polarization compared to the fiber diameter. The study also shows a potential role for MyD88 in regulating M1 BMMΦ signaling on the large vs. small fiber/pore size PDO scaffold. These data are instructive for the rationale design of implantable prosthetics designed to promote in situ regeneration.


Assuntos
Polaridade Celular/efeitos dos fármacos , Macrófagos/citologia , Polidioxanona/química , Polidioxanona/farmacologia , Engenharia Tecidual , Alicerces Teciduais/química , Animais , Arginase/metabolismo , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Meios de Cultivo Condicionados/farmacologia , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Endotoxinas/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/enzimologia , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Eletrônica de Varredura , Fator 88 de Diferenciação Mieloide/metabolismo , Neovascularização Fisiológica/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II/metabolismo , Fenótipo , Porosidade , Transdução de Sinais/efeitos dos fármacos
16.
Front Immunol ; 3: 117, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22593761

RESUMO

Mast cells are central players in immune surveillance and activation, positioned at the host-environment interface. Understanding the signaling events controlling mast cell function, especially those that maintain host homeostasis, is an important and still less understood area of mast cell-mediated disease. With respect to allergic disease, it is well established that IgE and its high affinity receptor FcεRI are major mediators of mast cell activation. However, IgG-mediated signals can also modulate mast cell activities. Signals elicited by IgG binding to its cognate receptors (FcγR) are the basis for autoimmune disorders such as lupus and rheumatoid arthritis. Using knowledge of IgE-mediated mast cell signaling, recent work has begun to illuminate potential overlap between FcεRI and FcγR signal transduction. Herein we review the importance of Src family kinases in FcεRI and FcγR signaling, the role of the transcription factor STAT5, and impingement of the regulatory cytokines IL-4, IL-10, and TGFß1 upon this network.

17.
J Immunol ; 188(9): 4360-8, 2012 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-22450804

RESUMO

Anaphylaxis is a rapid, life-threatening hypersensitivity reaction. Until recently, it was mainly attributed to histamine released by mast cells activated by allergen crosslinking (XL) of FcεRI-bound allergen-specific IgE. However, recent reports established that anaphylaxis could also be triggered by basophil, macrophage, and neutrophil secretion of platelet-activating factor subsequent to FcγR stimulation by IgG/Ag complexes. We have investigated the contribution of Fyn and Lyn tyrosine kinases to FcγRIIb and FcγRIII signaling in the context of IgG-mediated passive systemic anaphylaxis (PSA). We found that mast cell IgG XL induced Fyn, Lyn, Akt, Erk, p38, and JNK phosphorylation. Additionally, IgG XL of mast cells, basophils, and macrophages resulted in Fyn- and Lyn-regulated mediator release in vitro. FcγR-mediated activation was enhanced in Lyn-deficient (knockout [KO]) cells, but decreased in Fyn KO cells, compared with wild-type cells. More importantly, Lyn KO mice displayed significantly exacerbated PSA features whereas no change was observed for Fyn KO mice, compared with wild-type littermates. Intriguingly, we establish that mast cells account for most serum histamine in IgG-induced PSA. Taken together, our findings establish pivotal roles for Fyn and Lyn in the regulation of PSA and highlight their unsuspected functions in IgG-mediated pathologies.


Assuntos
Anafilaxia/imunologia , Imunoglobulina G/imunologia , Mastócitos/imunologia , Proteínas Proto-Oncogênicas c-fyn/imunologia , Quinases da Família src/imunologia , Alérgenos/genética , Alérgenos/imunologia , Anafilaxia/genética , Anafilaxia/patologia , Animais , MAP Quinases Reguladas por Sinal Extracelular/genética , MAP Quinases Reguladas por Sinal Extracelular/imunologia , Imunoglobulina G/genética , MAP Quinase Quinase 4/genética , MAP Quinase Quinase 4/imunologia , Mastócitos/patologia , Camundongos , Camundongos Knockout , Fosforilação/genética , Fosforilação/imunologia , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/imunologia , Proteínas Proto-Oncogênicas c-fyn/genética , Receptores de IgG/genética , Receptores de IgG/imunologia , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Quinases da Família src/genética
18.
J Neurooncol ; 106(3): 461-71, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21858729

RESUMO

Herein we continue the study of matrix metalloproteinase-1 (MMP-1) with respect to glioblastoma multiforme (GBM) cell tumorigenicity and angiogenesis. A model of tumorigenicity with cells stably altered to over-express or knock-down MMP-1 revealed that it significantly increases tumor incidence and size. Organized endothelial growth in human umbilical vein endothelial cell (HUVEC)-GBM co-cultures was significantly increased in the presence of MMP-1. CD31 analysis of model tumors elucidated a substantial recruitment of endothelium in MMP-1 enhanced samples. Antibody arrays indicated an inverse expression of certain anti-angiogenic factors with respect to MMP-1, the most notable of which was a significant increase in tissue inhibitor of metalloproteinases-4 (TIMP-4) in the absence of MMP-1, as validated by immunoblot.


Assuntos
Neoplasias Encefálicas/genética , Regulação Neoplásica da Expressão Gênica/genética , Glioblastoma/genética , Metaloproteinase 1 da Matriz/genética , Inibidores Teciduais de Metaloproteinases/metabolismo , Animais , Linhagem Celular Tumoral , Transformação Celular Neoplásica/genética , Técnicas de Cocultura , Modelos Animais de Doenças , Células Endoteliais/patologia , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Metaloproteinase 1 da Matriz/metabolismo , Camundongos , Camundongos Nus , Mutação/genética , Neovascularização Patológica/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Proteômica , Inibidores Teciduais de Metaloproteinases/genética , Veias Umbilicais/citologia , Inibidor Tecidual 4 de Metaloproteinase
19.
J Biol Chem ; 287(3): 2045-54, 2012 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-22130676

RESUMO

Previous studies indicate that STAT5 expression is required for mast cell development, survival, and IgE-mediated function. STAT5 tyrosine phosphorylation is swiftly and transiently induced by activation of the high affinity IgE receptor, FcεRI. However, the mechanism for this mode of activation remains unknown. In this study we observed that STAT5 co-localizes with FcεRI in antigen-stimulated mast cells. This localization was supported by cholesterol depletion of membranes, which ablated STAT5 tyrosine phosphorylation. Through the use of various pharmacological inhibitors and murine knock-out models, we found that IgE-mediated STAT5 activation is dependent upon Fyn kinase, independent of Syk, PI3K, Akt, Bruton's tyrosine kinase, and JAK2, and enhanced in the context of Lyn kinase deficiency. STAT5 immunoprecipitation revealed that unphosphorylated protein preassociates with Fyn and that this association diminishes significantly during mast cell activation. SHP-1 tyrosine phosphatase deficiency modestly enhanced STAT5 phosphorylation. This effect was more apparent in the absence of Gab2, a scaffolding protein that docks with multiple negative regulators, including SHP-1, SHP-2, and Lyn. Targeting of STAT5A or B with specific siRNA pools revealed that IgE-mediated mast cell cytokine production is selectively dependent upon the STAT5B isoform. Altogether, these data implicate Fyn as the major positive mediator of STAT5 after FcεRI engagement and demonstrate importantly distinct roles for STAT5A and STAT5B in mast cell function.


Assuntos
Citocinas/biossíntese , Mastócitos/metabolismo , Receptores de IgE/metabolismo , Fator de Transcrição STAT5/metabolismo , Transdução de Sinais/fisiologia , Proteínas Adaptadoras de Transdução de Sinal , Tirosina Quinase da Agamaglobulinemia , Animais , Células Cultivadas , Colesterol/genética , Colesterol/metabolismo , Citocinas/genética , Janus Quinase 2/genética , Janus Quinase 2/metabolismo , Mastócitos/citologia , Camundongos , Camundongos Knockout , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Fosforilação/fisiologia , Proteína Tirosina Fosfatase não Receptora Tipo 6/genética , Proteína Tirosina Fosfatase não Receptora Tipo 6/metabolismo , Proteínas Tirosina Quinases/genética , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-fyn/genética , Proteínas Proto-Oncogênicas c-fyn/metabolismo , Receptores de IgE/genética , Fator de Transcrição STAT5/genética , Quinases da Família src/genética , Quinases da Família src/metabolismo
20.
J Neurooncol ; 96(2): 201-9, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19629394

RESUMO

High grade gliomas invariably recur due in a large part to tumor cells permeating normal brain in an inaccessible, diffuse manner. Previous work demonstrates that the expression of matrix metalloproteinases (MMP) contributes to this characteristic. Not only can MMPs assist a cell in traversing its environment by clearing extracellular matrix molecules, but they can also impact non-traditional downstream signals that affect a cell's ability to interact and respond to its surroundings. Contributions to the induction of MMP expression and functional significance in glioma are still under investigation. Evidence in other cancer settings indicates that nitric oxide (NO) may play a role in tumor/cell progression that can influence MMP production. Matrix metalloproteinase-1 (MMP-1), also known as interstitial collagenase, and the constitutive nitric oxide synthases (NOS) have been shown to be over-expressed in high grade gliomas. In the current study we investigated the potential involvements of NO with regard to MMP-1 and functional glioma cell movement. With the treatment of the NO donor sodium nitroprusside (SNP), there was significant induction of MMP-1 mRNA, secreted MMP-1 protein and motility of glioma cell lines within 48 h. RNA inhibition of MMP-1 through transient transfection of three MMP-1 specific siRNAs revealed a marked abrogation of the NO-mediated induction of motility. In addition, application of the NOS inhibitor N(omega)-Nitro-L-arginine methyl ester (L-NAME) impaired movement of glioma cells. These data provide evidence for a regulatory axis of high grade glioma cell movement from NO through MMP-1, with NOS inhibitor results showing promise for future pharmacologic investigation.


Assuntos
Movimento Celular/fisiologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioma/metabolismo , Glioma/fisiopatologia , Metaloproteinase 1 da Matriz/metabolismo , Óxido Nítrico/metabolismo , Análise de Variância , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Humanos , Metaloproteinase 1 da Matriz/genética , NG-Nitroarginina Metil Éster/farmacologia , Doadores de Óxido Nítrico/farmacologia , Nitroprussiato/farmacologia , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/metabolismo , Transfecção/métodos
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