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1.
Zhongguo Zhong Yao Za Zhi ; 45(2): 361-366, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-32237319

RESUMO

To investigate the effects of butyl alcohol extract of Baitouweng Decoction(BAEB) on neutrophil chemotaxis in vaginal mucosa of mice with vulvovaginal candidiasis(VVC). Seventy-two SPF female Kunming mice were randomly divided into normal control group, model group, fluconazole group, BAEB low-dose group, middle-dose group and high-dose group. Subcutaneous injection of estradiol benzoate was conducted to induce pseudo-estrus, and then 2×10~6 CFU·mL~(-1)of Candida albicans was inoculated into vaginal lumen, followed by drug treatment for 7 days. Gram staining was used to observe the morphological changes of C. albicans in vagina; vaginal fungal load was detected on agar plate. Histological changes of vaginal tissues in mice were observed by HE staining. Lactate dehydrogenase(LDH), interleukin-6(IL-6) and tumor necrosis factor(TNF-α) levels in mouse lavage fluid were detected by enzyme-linked immunosorbent assay(ELISA). Neutrophils in vaginal lavage fluid was observed and counted by using Pap smear. The levels of IL-8 and MIP-2 in vaginal mucosa were detected by ELISA. IL-8 and MIP-2 mRNA levels in vaginal mucosa of mice were detected by qRT-PCR. The results showed that as compared with the normal group, VVC model group had a large number of hyphae and a high level of fungal loadinvagina. The vaginal mucosa was completely destroyed, the number of neutrophils increased, and the protein and mRNA levels of IL-8 and MIP-2 were up-regulated. After BAEB treatment, the hyphae of the treatment group was decreased, the fungal load was decreased, the impaired mucosa showed different degrees of improvement, the inflammatory factors were decreased to varying degrees, and the protein and mRNA levels of chemokine IL-8 and MIP-2 were down-regulated. In conclusion, BAEB may be used to treat VVC by inhibiting vulvovaginal candidiasis via blocking neutrophils recruitment into vagina.


Assuntos
Candidíase Vulvovaginal/tratamento farmacológico , Quimiotaxia/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Neutrófilos/efeitos dos fármacos , 1-Butanol , Animais , Candida albicans , Feminino , Camundongos , Membrana Mucosa/citologia , Membrana Mucosa/efeitos dos fármacos , Neutrófilos/citologia , Vagina/citologia , Vagina/diagnóstico por imagem
2.
Int J Pediatr Otorhinolaryngol ; 128: 109699, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31614241

RESUMO

OBJECTIVE: The middle ear epithelium is derived from the neural crest and endoderm, which line distinct regions of the middle ear cavity. In this study, we investigated the localization of stem/progenitor cells in the middle ear mucosa of adult mice and the effects of keratinocyte growth factor (KGF) on the cell kinetics of stem/progenitor cells in vivo. METHODS: In this study, after KGF-expression vector was transfected in the ear, two kinds of thymidine analogues, BrdU and EdU, were transferred at different time points. BrdU was detected by immunohistochemistry and EdU was detected by click chemistry. We also performed immunohistochemistry using anti-Keratin14 (K14) antibody (an undifferentiated epithelial cell marker), anti-p63 antibody (a stem/progenitor cell marker) and anti-acetylated α-tubulin antibody (a ciliated epithelial cell marker). RESULTS: A large number of EdU-positive cells were detected in the thickened mucosal epithelium of the pars flaccida and attic region at Day 1 after KGF transfection. Interestingly, in the mucosal epithelium overlying the promontory of the cochlea, many EdU-positive cells were detected. These cells were also positive for K14 and p63. The acetylated α-tubulin positive cells were reduced in the attic region at Day 1 after KGF transfection. CONCLUSION: These findings indicate that KGF over-expression may increase stem/progenitor cell proliferation in the mucosal epithelium not only within the attic which is typical in middle ear cholesteatoma, but also overlying the promontory of the cochlea.


Assuntos
Orelha Média/citologia , Células Epiteliais/metabolismo , Fator 7 de Crescimento de Fibroblastos/metabolismo , Membrana Mucosa/citologia , Células-Tronco/fisiologia , Animais , Proliferação de Células , Orelha Média/metabolismo , Fator 7 de Crescimento de Fibroblastos/genética , Queratina-14/metabolismo , Masculino , Camundongos , Membrana Mucosa/metabolismo , Transativadores/metabolismo , Transfecção , Tubulina (Proteína)/metabolismo
3.
Anat Histol Embryol ; 49(1): 31-37, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31571240

RESUMO

The distribution of glucagon-like peptide 1 (GLP-1)-positive cells in digestive tracts and pancreases of aquatic vertebrates was investigated by immunohistochemical staining method. The results suggested that GLP-1-positive cells were distributed in the columnar mucous epithelium and tubular glands of lamina propria in the digestive system. However, GLP-1-positive cells were also found in subepithelial lamina propria of the mucosae and muscularis in each segment of the digestive tract of Rana nigromaculata. The distribution densities of these cells reached peaks in the stomachs, and the middle or end segments of small intestines of Chinese softshell turtle, Bufo gargarizans, R. nigromaculata and catfish, and there was the third distribution density peak in the rectum of catfish. The total amount or overall density of GLP-1-positive cells varied a lot in the digestive tracts of different animal species. The distribution density was relatively low in the digestive tract of chub and reached the maximum in the digestive tracts of snakehead and catfish, but no GLP-1-positive cells were found in the digestive tract of bighead carp. GLP-1-positive cells were densely distributed in the pancreases of Chinese softshell turtle, B. gargarizans and R. nigromaculata. These cells spread over the superficial layers of islets or scattered in exocrine pancreas in the pancreas of B. gargarizans, spread in the endocrine cells or scattered in the pancreas of Chinese softshell turtle, scattered in the pancreas of R. nigromaculata and distributed in the superficial layers of islets in the pancreas of catfish.


Assuntos
Sistema Digestório/citologia , Sistema Digestório/metabolismo , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Anfíbios/metabolismo , Animais , Organismos Aquáticos/metabolismo , Células Epiteliais/metabolismo , Peixes/metabolismo , Imuno-Histoquímica , Membrana Mucosa/citologia , Membrana Mucosa/metabolismo , Pâncreas/citologia , Pâncreas/metabolismo , Tartarugas/metabolismo , Vertebrados/metabolismo
4.
Adv Gerontol ; 32(4): 606-613, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31800191

RESUMO

The structure and cytoarchitecture of the structures of the wall of the esophagus was studied by histological and morphometric methods. We investigated the upper, middle and lower third of the esophagus in people 1 and 2 of the mature, elderly and senile age. It was established that in old age comes the destruction of the epithelial lining, the muscular propria of the mucous membrane. The number of lymphocytes, neutrophilic leukocytes and eosinophils in the epithelium increases by 2-3 times. The content of cells of the fibroblastic and lymphoid series increases in lamina propria and glands. Infiltrates appear in the mucous membrane, microerosions appear. Particularly vulnerable areas are the papillas of lamina propria. In the places of their location the most frequent accumulations of lymphocytes are noted. Among mast cells, the processes of degranulation increase, the number of mast cells with a small amount of granules increases. Inflammation is often found in the lower sections. Age-related changes in the wall of the esophagus depend on the individual characteristics of a person.


Assuntos
Células Epiteliais , Esôfago , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Envelhecimento , Células Epiteliais/citologia , Células Epiteliais/patologia , Epitélio/patologia , Esôfago/citologia , Esôfago/patologia , Humanos , Pessoa de Meia-Idade , Membrana Mucosa/citologia , Membrana Mucosa/patologia
5.
PLoS One ; 14(12): e0226343, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31869348

RESUMO

The oral, cervical, and genital mucosa, covered by stratified squamous epithelia with polarized organization and strong tight and adherens junctions, play a critical role in preventing transmission of viral pathogens, including human immunodeficiency virus (HIV). HIV-1 interaction with mucosal epithelial cells may depolarize epithelia and disrupt their tight and adherens junctions; however, the molecular mechanism of HIV-induced epithelial disruption has not been completely understood. We showed that prolonged interaction of cell-free HIV-1 virions, and viral envelope and transactivator proteins gp120 and tat, respectively, with tonsil, cervical, and foreskin epithelial cells induces an epithelial-mesenchymal transition (EMT). EMT is an epigenetic process leading to the disruption of mucosal epithelia and allowing the paracellular spread of viral and other pathogens. Interaction of cell-free virions and gp120 and tat proteins with epithelial cells substantially reduced E-cadherin expression and activated vimentin and N-cadherin expression, which are well-known mesenchymal markers. HIV gp120- and tat-induced EMT was mediated by SMAD2 phosphorylation and activation of transcription factors Slug, Snail, Twist1 and ZEB1. Activation of TGF-ß and MAPK signaling by gp120, tat, and cell-free HIV virions revealed the critical roles of these signaling pathways in EMT induction. gp120- and tat-induced EMT cells were highly migratory via collagen-coated membranes, which is one of the main features of mesenchymal cells. Inhibitors of TGF-ß1 and MAPK signaling reduced HIV-induced EMT, suggesting that inactivation of these signaling pathways may restore the normal barrier function of mucosal epithelia.


Assuntos
Células Epiteliais/efeitos dos fármacos , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Genitália/citologia , Proteína gp120 do Envelope de HIV/farmacologia , Mucosa Bucal/efeitos dos fármacos , Produtos do Gene tat do Vírus da Imunodeficiência Humana/farmacologia , Células Cultivadas , Pré-Escolar , Células Epiteliais/citologia , Células Epiteliais/fisiologia , Feminino , Genitália/virologia , Células HEK293 , Proteína gp120 do Envelope de HIV/metabolismo , HIV-1/metabolismo , Humanos , Lactente , Recém-Nascido , Queratinócitos/efeitos dos fármacos , Queratinócitos/fisiologia , Masculino , Mucosa Bucal/fisiologia , Membrana Mucosa/citologia , Membrana Mucosa/efeitos dos fármacos
6.
J Immunol Methods ; 474: 112665, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31525366

RESUMO

Mucosal tissues are enriched in γδ T lymphocytes, which maintain epithelial homeostasis, however, the homeostatic mechanisms are still incompletely understood. To elucidate their role in the tissue integrity governance within the female genital mucosa we employed flow cytometry, which is a powerful tool used for the characterization of tissue-resident immune cells, however, often requiring cell release upon tissue enzymatic disaggregation. Here, we analyzed the impact of various proteolytic enzymes in their ability to effectively isolate viable immune cells from the reproductive system of non-pregnant mice. Murine vaginas and uteri were digested using commercially available enzyme blends (liberases) and single enzymes (dispase II and collagenase IV). Among tested enzymes, liberases released the highest number of cells from digested tissues while dispase II and collagenase IV led to a significant decrease in the number of isolated live cells. Also, liberases had only minor detrimental effects on cell viability and detection of CD45, CD3ε, γδ TCR and CD11c positive cells. We found that a single liberase blend called Liberase TL was the most suited for the analysis of γδ T cells in the reproductive tract. By examining two distinct phases of the estrous cycle - estrus and diestrus, characterized by high and low epithelial stratification, respectively, we showed that higher numbers of γδ T lymphocytes were present in the latter cycle phase in vagina and uterus. Interestingly, the diestrus-associated increase in γδ T lymphocyte number was also observed in reproductive tract draining lumbar lymph nodes but not in more distant, inguinal lymph nodes. Our data indicate that enzymes used for reproductive mucosa digestion have profound effects on the cell viability and isolation efficiency, which consequently influence the phenotypic and quantitative analysis of immune cells.


Assuntos
Separação Celular , Imunidade nas Mucosas , Membrana Mucosa/imunologia , Peptídeo Hidrolases/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Subpopulações de Linfócitos T/imunologia , Útero/imunologia , Vagina/imunologia , Animais , Colagenases/metabolismo , Diestro/imunologia , Endopeptidases/metabolismo , Estro/imunologia , Feminino , Citometria de Fluxo , Contagem de Linfócitos , Camundongos Endogâmicos C57BL , Membrana Mucosa/citologia , Fenótipo , Termolisina/metabolismo , Útero/citologia , Vagina/citologia
7.
In Vivo ; 33(5): 1499-1505, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31471398

RESUMO

BACKGROUND/AIM: The mouse vagina exhibits stratified squamous epithelium, which is comprised of multiple cell layers. We previously showed that erbB signaling, induced by epithelial estrogen receptor 1 (ESR1), is required for the initial differentiation of the epithelium. However, the downstream effector that mediates terminal differentiation in the apical layers remains elusive. The contribution of fibroblast growth factor (FGF) to vaginal epithelial cell differentiation was investigated. MATERIALS AND METHODS: Vaginas from wild-type or epithelium-specific Esr1 conditional knockout (cKO) mice were analyzed using immunohistochemistry and quantitative real-time RT-PCR. RESULTS: Of the FGF ligands examined, Fgf22 mRNA was significantly induced following estrogen treatment. Furthermore, FGF downstream signaling, phosphorylated FRS2 and ERK1/2 were exclusively expressed in the apical layers of the vaginal epithelium. No changes in such expression were observed in the Esr1 cKO mice. CONCLUSION: FGF-ERK/MAPK pathway may be a main inducer of terminal differentiation in the mouse vaginal epithelium.


Assuntos
Diferenciação Celular , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Membrana Mucosa/citologia , Membrana Mucosa/metabolismo , Vagina , Animais , Biomarcadores , Diferenciação Celular/genética , Feminino , Ligantes , Camundongos , Camundongos Transgênicos , Transdução de Sinais
8.
Neurourol Urodyn ; 38(8): 2093-2103, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31338895

RESUMO

To describe and illustrate the structure of the propria, the bladder of adult rats was fixed in controlled conditions of distension and examined by light and electron microscopy. The lamina propria, ~50 µm thick in the distended bladder, consists of a superficial part (the cellular component), adjacent to the urothelium, rich in nerves, capillaries, fibroblasts and thin bundles of collagen, and a deep, thicker part (the fibrous component), adjacent to the detrusor, rich in large collagen fibres and with few fibroblasts. In the cellular part there is an extensive plexus of afferent nerve fibers and a dense capillary network (with numerous pericytes), lying close to the urothelium, that is unique to the bladder. The main resident cells are fibroblasts, adhering to each other at the end of laminar extensions without forming specialized junctions. The deep part of the lamina propria is made of thick collagen fibers, interwoven and crisscrossing each other, with a few fibroblasts in the interstitial spaces between them. In summary, the superficial part of the lamina propria has most of the bladder afferent nerves, contains many fibroblasts and has a network of suburothelial capillaries. The deep part as a whole forms an ovoid balloon of woven fibrous material that is acted upon by the detrusor musculature attached to its outer surface. The lamina propria is a strong fibrous barrier between urothelium and musculature. The abundance of collagen points to the main role for its fibroblasts, that is, the production of collagen fibrils, assisting the mechanical role of the lamina propria.


Assuntos
Membrana Mucosa/ultraestrutura , Bexiga Urinária/ultraestrutura , Animais , Capilares/citologia , Capilares/ultraestrutura , Fibroblastos/citologia , Fibroblastos/ultraestrutura , Masculino , Microscopia Eletrônica , Membrana Mucosa/citologia , Fibras Nervosas , Ratos , Bexiga Urinária/citologia , Urotélio/citologia , Urotélio/ultraestrutura
9.
Nat Commun ; 10(1): 2892, 2019 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-31253778

RESUMO

Clinical manifestations and response to therapies in ulcerative colitis (UC) are heterogeneous, yet patient classification criteria for tailored therapies are currently lacking. Here, we present an unsupervised molecular classification of UC patients, concordant with response to therapy in independent retrospective cohorts. We show that classical clustering of UC patient tissue transcriptomic data sets does not identify clinically relevant profiles, likely due to associated covariates. To overcome this, we compare cross-sectional human data sets with a newly generated longitudinal transcriptome profile of murine DSS-induced colitis. We show that the majority of colitis risk-associated gene expression peaks during the inflammatory rather than the recovery phase. Moreover, we achieve UC patient clustering into two distinct transcriptomic profiles, differing in neutrophil-related gene activation. Notably, 87% of patients in UC1 cluster are unresponsive to two most widely used biological therapies. These results demonstrate that cross-species comparison enables stratification of patients undistinguishable by other molecular approaches.


Assuntos
Mapeamento Cromossômico , Colite/metabolismo , Doenças Inflamatórias Intestinais/metabolismo , Transcriptoma , Animais , Colite/induzido quimicamente , Colite/genética , Citometria de Fluxo , Regulação da Expressão Gênica , Humanos , Camundongos , Membrana Mucosa/citologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
10.
Nat Commun ; 10(1): 2214, 2019 05 17.
Artigo em Inglês | MEDLINE | ID: mdl-31101810

RESUMO

CD8+ T cells provide a critical defence from pathogens at mucosal epithelia including the female reproductive tract (FRT). Mucosal immunisation is considered essential to initiate this response, however this is difficult to reconcile with evidence that antigen delivered to skin can recruit protective CD8+ T cells to mucosal tissues. Here we dissect the underlying mechanism. We show that adenovirus serotype 5 (Ad5) bio-distributes at very low level to non-lymphoid tissues after skin immunisation. This drives the expansion and activation of CD3- NK1.1+ group 1 innate lymphoid cells (ILC1) within the FRT, essential for recruitment of CD8+ T-cell effectors. Interferon gamma produced by activated ILC1 is critical to licence CD11b+Ly6C+ monocyte production of CXCL9, a chemokine required to recruit skin primed CXCR3+ CD8+T-cells to the FRT. Our findings reveal a novel role for ILC1 to recruit effector CD8+ T-cells to prevent virus spread and establish immune surveillance at barrier tissues.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Genitália Feminina/imunologia , Pele/imunologia , Vacinas Virais/administração & dosagem , Viroses/prevenção & controle , Adenovírus Humanos/genética , Adenovírus Humanos/imunologia , Administração Cutânea , Animais , Quimiocina CXCL9 , Modelos Animais de Doenças , Feminino , Genitália Feminina/citologia , Genitália Feminina/virologia , Células HEK293 , Interações Hospedeiro-Patógeno/imunologia , Humanos , Imunidade Inata , Camundongos , Camundongos Endogâmicos C57BL , Monócitos/imunologia , Membrana Mucosa/citologia , Membrana Mucosa/imunologia , Membrana Mucosa/virologia , Receptores CXCR3 , Pele/citologia , Pele/virologia , Resultado do Tratamento , Vacinação/métodos , Vacinas Sintéticas/administração & dosagem , Vacinas Sintéticas/genética , Vacinas Sintéticas/imunologia , Vacinas Virais/genética , Vacinas Virais/imunologia , Viroses/imunologia , Viroses/virologia , Produtos do Gene gag do Vírus da Imunodeficiência Humana/genética , Produtos do Gene gag do Vírus da Imunodeficiência Humana/imunologia
11.
mBio ; 10(3)2019 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-31088924

RESUMO

Trichomonas vaginalis, a prevalent sexually transmitted parasite, adheres to and induces cytolysis of human mucosal epithelial cells. We have characterized a hypothetical protein, TVAG_393390, with predicted tertiary structure similar to that of mammalian cadherin proteins involved in cell-cell adherence. TVAG_393390, renamed cadherin-like protein (CLP), contains a calcium-binding site at a position conserved in cadherins. CLP is surface localized, and its mRNA and protein levels are significantly upregulated upon parasite adherence to host cells. To test the roles of CLP and its calcium-binding dependency during host cell adherence, we first demonstrated that wild-type CLP (CLP) binds calcium with a high affinity, whereas the calcium-binding site mutant protein (CLP-mut) does not. CLP and CLP-mut constructs were then used to overexpress these proteins in T. vaginalis Parasites overexpressing CLP have ∼3.5-fold greater adherence to host cells than wild-type parasites, and this increased adherence is ablated by mutating the calcium-binding site. Additionally, competition with recombinant CLP decreased parasite binding to host cells. We also found that overexpression of CLP induced parasite aggregation which was further enhanced in the presence of calcium, whereas CLP-mut overexpression did not affect aggregation. Lastly, parasites overexpressing wild-type CLP induced killing of host cells ∼2.35-fold, whereas parasites overexpressing CLP-mut did not have this effect. These analyses describe the first parasitic CLP and demonstrate a role for this protein in mediating parasite-parasite and host-parasite interactions. T. vaginalis CLP may represent convergent evolution of a parasite protein that is functionally similar to the mammalian cell adhesion protein cadherin, which contributes to parasite pathogenesis.IMPORTANCE The adherence of pathogens to host cells is critical for colonization of the host and establishing infection. Here we identify a protein with no known function that is more abundant on the surface of parasites that are better at binding host cells. To interrogate a predicted function of this protein, we utilized bioinformatic protein prediction programs which allowed us to uncover the first cadherin-like protein (CLP) found in a parasite. Cadherin proteins are conserved metazoan proteins with central roles in cell-cell adhesion, development, and tissue structure maintenance. Functional characterization of this CLP from the unicellular parasite Trichomonas vaginalis demonstrated that the protein mediates both parasite-parasite and parasite-host adherence, which leads to an enhanced killing of host cells by T. vaginalis Our findings demonstrate the presence of CLPs in unicellular pathogens and identify a new host cell binding protein family in a human-infective parasite.


Assuntos
Caderinas/genética , Células Epiteliais/metabolismo , Proteínas de Protozoários/metabolismo , Trichomonas vaginalis/patogenicidade , Caderinas/metabolismo , Cálcio/metabolismo , Adesão Celular , Linhagem Celular , Células Epiteliais/parasitologia , Feminino , Humanos , Membrana Mucosa/citologia , Domínios Proteicos , Proteínas de Protozoários/genética , Ativação Transcricional , Regulação para Cima
12.
Immunol Lett ; 210: 1-9, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30904566

RESUMO

Tuft cells are epithelial chemosensory cells with unique morphological and molecular characteristics, the most noticeable of which is a tuft of long and thick microvilli on their apical side, as well as expression of a very distinct set of genes, including genes encoding various members of the taste transduction machinery and pro-inflammatory cyclooxygenases. Initially discovered in rat trachea, tuft cells were gradually identified in various mucosal tissues, and later also in non-mucosal tissues, most recent of which is the thymus. Although tuft cells were discovered more than 60 years ago, their functions in the various tissues remained a mystery until recent years. Today, tuft cells are thought to function as sensors of various types of chemical signals, to which they respond by secretion of diverse biological mediators such as IL25 or acetylcholine. Intestinal tuft cells were also shown to mediate type 2 immunity against parasites. Here, we review the current knowledge on tuft cell characteristics, development and heterogeneity, discuss their potential functions and explore the possible implications and significance of their discovery in the thymus.


Assuntos
Células Epiteliais/citologia , Células Epiteliais/metabolismo , Membrana Mucosa/citologia , Timo/citologia , Animais , Biomarcadores , Diferenciação Celular , Suscetibilidade a Doenças , Humanos , Microvilosidades/metabolismo , Fenótipo
13.
Hum Vaccin Immunother ; 15(6): 1409-1420, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30836838

RESUMO

CD4+ and CD8+ T subsets are essential components of the adaptive immune system which act in concert at the site of infections to effectively protect against pathogens. Very limited data is available in humans regarding the relationship between CD4+ and CD8+ S. Typhi responsive cells in the terminal ileum mucosa (TI) and peripheral blood following Ty21a oral typhoid immunization. Here, we compared TI lamina propria mononuclear cells (LPMC) and peripheral blood CD4+ and CD8+ T memory (TM) subsets responses and their relationship by Spearman's correlation following Ty21a immunization in volunteers undergoing routine colonoscopy. We observed that Ty21a immunization (i) influences the homing and accumulation of both CD4+ and CD8+ T cells in the TI, particularly integrin α4ß7+ CCR9+ CD8+ T cells, (ii) elicits significantly higher frequencies of LPMC S. Typhi-responsive CD8+ T multifunctional (CD107a, IFNγ, IL-17A and/or MIP1ß) cells than their CD4+ T counterparts, and (iii) results in the correlation of LPMC CD4+ Teffector/memory (TEM) S. Typhi responses (CD107a, IFNγ, TNFα, IL-17A and/or MIP1ß) to their LPMC CD8+ TEM counterparts. Moreover, we demonstrated that these positive correlations between CD4+ and CD8+ TEM occur primarily in TI LPMC but not in PBMC, suggesting important differences in responses between the mucosal and systemic compartments following oral Ty21a immunization. This study provides the first demonstration of the correlation of S. Typhi-specific CD4+ and CD8+ TM responses in the human terminal ileum mucosa and provides valuable information regarding the generation of mucosal and systemic immune responses following oral Ty21a-immunization which might impact future vaccine design and development.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Memória Imunológica , Leucócitos Mononucleares/imunologia , Membrana Mucosa/imunologia , Polissacarídeos Bacterianos/imunologia , Vacinas Tíficas-Paratíficas/imunologia , Administração Oral , Idoso , Feminino , Humanos , Íleo/imunologia , Imunização , Masculino , Pessoa de Meia-Idade , Membrana Mucosa/citologia , Polissacarídeos Bacterianos/administração & dosagem , Salmonella typhi , Vacinas Tíficas-Paratíficas/administração & dosagem
14.
Microbiol Spectr ; 7(2)2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30848233

RESUMO

Over the past few decades, in vitro cell culture systems have greatly expanded our understanding of host-pathogen interactions. However, studies using these models have been limited by the fact that they lack the complexity of the human body. Therefore, recent efforts that allow tissue architecture to be mimicked during in vitro culture have included the development of methods and technology that incorporate tissue structure, cellular composition, and efficient long-term culture. These advances have opened the door for the study of pathogens that previously could not be cultured and for the study of pathophysiological properties of infection that could not be easily elucidated using traditional culture models. Here we discuss the latest studies using organoids and engineering technology that have been developed and applied to the study of host-pathogen interactions in mucosal tissues.


Assuntos
Técnicas de Cultura de Células/métodos , Membrana Mucosa/citologia , Membrana Mucosa/microbiologia , Engenharia Tecidual/métodos , Animais , Interações Hospedeiro-Patógeno , Humanos , Intestinos/citologia , Intestinos/imunologia , Organoides/citologia , Organoides/microbiologia , Tecidos Suporte/microbiologia , Tropismo
16.
J Nutr ; 149(2): 344-353, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30721975

RESUMO

BACKGROUND: Several types of oligosaccharides are used in infant formula to improve the gut microbiota of formula-fed infants. We previously reported that a combination of 3 oligosaccharides (lactulose, raffinose, and galacto-oligosaccharides; LRG) and Bifidobacterium breve effectively increased B. breve numbers, acetate, and the expression of several immune- and gut hormone-related mRNAs in neonatal mice gut. OBJECTIVE: We investigated whether changes in neonatal gut microbiota alter gut immune and endocrine development. METHODS: We first compared postnatal day (PD) 14 with PD21 in C57BL/6J male mouse pups to identify the physiologic immune and endocrine changes during development. In a separate study, we administered phosphate-buffered saline (control group; CON), B. breve M-16V (M-16V), or M-16V + LRG to male mouse pups from PD6 to PD13, and analyzed the gut microbiota and immune and endocrine parameters on PD14 to evaluate whether M-16V + LRG accelerates gut immune and endocrine development. RESULTS: The proportion of regulatory T (Treg) cells in the CD4+ cells of large intestinal lamina propria lymphocytes (LPLs) was significantly increased (63% higher) at PD21 compared with PD14. The serum glucagon-like peptide (GLP)-1 tended to be lower (P = 0.0515) and that of GLP-2 was significantly lower (58% lower) at PD21 than at PD14. M-16V + LRG significantly increased the Treg proportion in large intestinal LPL CD4+ cells (20% and 29% higher compared with CON and M-16V, respectively) at PD14. M-16V + LRG also caused significant changes in expression of large intestinal mRNAs that are consistent with developmental progression, and increased serum concentrations of GLP-1 (207% and 311% higher compared with CON and M-16V, respectively) and GLP-2 (57% and 97% higher compared with CON and M-16V, respectively) at PD14. CONCLUSIONS: Neonatal administration of M-16V + LRG alters the gut microbiota and enhances gut immune and endocrine development in suckling mice.


Assuntos
Bifidobacterium breve , Intestinos/imunologia , Intestinos/fisiologia , Oligossacarídeos/farmacologia , Prebióticos/administração & dosagem , Probióticos/farmacologia , Animais , Animais Recém-Nascidos , Linfonodos/citologia , Linfócitos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Membrana Mucosa/citologia , Probióticos/administração & dosagem , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Linfócitos T Reguladores
17.
J Fish Biol ; 94(4): 648-659, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30762233

RESUMO

The objective of the present study was to describe the histology and histochemistry of the mucosal layer of the digestive tube of Piaractus brachypomus, and the histopathology associated with parasitism by Neoechinorhynchus sp. The digestive tube of P. brachypomus consists of three macroscopically distinct portions: short, rectilinear and elastic-walled ooesophagus, J-shaped siphon stomach and a long intestine with rectilinear and curved portions, defined by patterns of villi as foregut, midgut, and hindgut. Histological and histochemical differences were observed in the mucosal layers of the different digestive tube regions, such as intense production of neutral and acidic mucous substances in the pseudostratified mucosal epithelium of the oesophagus; positive periodic acid Schiff reagent (PAS)reactions at the apex of the columnar epithelial cells of the stomach and increased intensity of histochemical reactions in the hindgut region. Neoechinorhynchus sp. was present in 85.7% of specimens examined, with a mean intensity of 7.4 ± 6.2 (±) and abundance of 6.33. Good health of the fish indicated by high relative condition factor values ( Kn ) and occurrence of only mild to moderate alteration in the mucosal layer indicated that Neoechinorhynchus sp. exhibits low pathogenicity towards P. brachypomus hosts in farming environments, with low levels of infection.


Assuntos
Acantocéfalos/fisiologia , Caraciformes/parasitologia , Doenças dos Peixes/patologia , Trato Gastrointestinal/anatomia & histologia , Helmintíase Animal/patologia , Animais , Caraciformes/anatomia & histologia , Esôfago/anatomia & histologia , Doenças dos Peixes/parasitologia , Trato Gastrointestinal/parasitologia , Histocitoquímica , Interações Hospedeiro-Parasita , Intestinos/anatomia & histologia , Membrana Mucosa/citologia , Membrana Mucosa/metabolismo , Membrana Mucosa/parasitologia , Estômago/anatomia & histologia
18.
PLoS One ; 14(2): e0212075, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30763359

RESUMO

HIV infection causes the progressive depletion of CD4+ T-lymphocytes and profound modifications of T-cell homeostasis, which persist despite virologically-suppressive treatment and have been linked to a worse clinical outcome. Enduring alterations of the gastrointestinal tract may represent the underlying pathogenic mechanisms of these phenomena. Twenty-six HIV-infected subjects were assessed over a 12-month period following the introduction of antiretroviral therapy. 18 uninfected individuals were enrolled as controls. Parameters of peripheral T-cell homeostasis (activation, maturation), gastrointestinal function (microbial translocation, gut inflammation, fecal microbiota composition) and mucosal immunity (CD4+CCR6+CD161+, CD4+CCR9+α4ß7+, stem cell memory CD4+/CD8+ T-cells) were assessed. CD4+CCR6+CD161+ cells were depleted in HIV-infected untreated subjects and maintained significantly lower levels compared to controls, despite the introduction of effective antiviral treatment. The frequency of gut-homing CD4+CCR9+α4ß7+ cells was also impaired in untreated infection and correlated with the HIV RNA load and CD4+HLADR+CD38+; during therapy, we observed a contraction of this pool in the peripheral blood and the loss of its correlation with antigenic exposure/immune activation. A partial correction of the balance between stem cell memory pools and T-cell homeostasis was registered following treatment. In HIV-infected subjects with moderate immune-suppression, antiretroviral therapy has a marginal impact on mucosal immune populations which feature distinctive kinetics in the periphery, possibly reflecting their diverse recruitment from the blood to the mucosa. The persistent defects in mucosal immunity may fuel peripheral T-cell abnormalities through diverse mechanisms, including the production of IL-17/IL-22, cellular permissiveness to infection and regulation of T-lymphocyte maturation.


Assuntos
Fármacos Anti-HIV/uso terapêutico , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Tolerância Imunológica , Imunidade nas Mucosas/efeitos dos fármacos , Membrana Mucosa/citologia , Adulto , Fármacos Anti-HIV/farmacologia , Interações Medicamentosas , Fezes/microbiologia , Feminino , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/imunologia , Trato Gastrointestinal/microbiologia , Infecções por HIV/microbiologia , Humanos , Masculino , Microbiota/efeitos dos fármacos , Pessoa de Meia-Idade , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia
19.
Inflammation ; 42(2): 682-689, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30406462

RESUMO

Several biologic processes affect the supporting peri-implant tissue leading to implant failure and complications, mainly referred to inflammation that is still poorly investigated in the peri-implant soft tissues. Our aim was to investigate in peri-implant healthy mucosa, peri-implant mucositis, and peri-implantitis the expression of some angiogenesis markers highly associated with inflammation, and evaluate its relationships with age, smoking, peri-implant pocket depth (PPD), and body max index (BMI). Moreover, we wanted to study the impact of these clinical parameters in the disease pathogenesis. Forty-eight total patients were recruited. Sixteen had at least one successfully osteointegrated dental implant (group A) and 32 had at least one osseointegrated implant in need of a peri-implant treatment for inflammatory/infectiveous reasons: precisely 16 for mucositis (group B) and 16 for peri-implantitis (group C). VEGF, CD34, and CD44 immunohistochemical expression was evaluated in the interproximal biopsies of marginal peri-implant tissue and correlated with the clinical parameters. A significant difference between groups in mean PPD was found, while the distribution by age, gender, smoking, and BMI resulted similar. Group C had significantly higher levels of VEGF, CD34, and CD44 expression compared to the other groups. VEGF, CD34, CD44, and peri-implant pocket depth were all positively correlated. Our study revealed that peri-implantitis is a condition characterized by unique and distinctive features. Our results supported that PPD has a great impact on the peri-implantitis and it is closely related to the inflammation marker expression. The identification of specific biomarkers might help in choosing distinct treatment approaches for target individuals.


Assuntos
Receptores de Hialuronatos/sangue , Inflamação/sangue , Microvasos , Mucosite/sangue , Membrana Mucosa , Peri-Implantite/sangue , Fator A de Crescimento do Endotélio Vascular/sangue , Adulto , Fatores Etários , Biomarcadores/sangue , Feminino , Humanos , Inflamação/diagnóstico , Masculino , Pessoa de Meia-Idade , Membrana Mucosa/irrigação sanguínea , Membrana Mucosa/citologia , Obesidade , Fatores de Risco , Fumar
20.
Front Immunol ; 9: 2116, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30294324

RESUMO

In mammals, M cells can take up antigens through mucosal surfaces of the gut and the respiratory tract. Since M cells are deficient of lysosomes and phagosomes, the antigens are directly delivered to the mucosa-associated lymphoid tissue (MALT) without degradation. In teleost fish, the entire body surface (gills, skin, and intestinal system) is covered by mucus; however, specific antigen-sampling cells have not yet been identified in their mucosal tissues. Here, we show that two phenotypes of antigen-sampling cells take up antigens through epithelial surfaces of the rainbow trout gill. One phenotype of antigen-sampling cells has features of monocyte/macrophage/dendritic cell-type cells; they have large vacuoles in the cytoplasm and express PTPRC (CD45), CD83, IL-1ß, and IL-12p40b. The second phenotype exhibits similar characteristics to mammalian M cells; the corresponding cells bind the lectin UEA-1 but not WGA and show expression of M cell marker gene Anxa5. In contrast to mammalian M cells, teleost M-type cells were found to exhibit small vacuoles in their cytoplasm and to express almost all genes related to the "phagosome", "lysosome," and "antigen processing and presentation" pathways. Furthermore, MHC class II was constitutively expressed on a fraction of M-type cells, and this expression was significantly increased after antigen uptake, suggesting that the MHC class II is inducible by antigen stimulation. Here, we suggest that teleost M-type cells play a role in the phylogenetically primitive teleost immune system, similar to bona-fide M cells. In addition, the presence of MHC class II expression suggests an additional role in antigen presentation in the gills, which are an organ with high T cell abundance, especially in interbranchial lymphoid tissue. The present results suggest an unconventional antigen presentation mechanism in the primitive mucosal immune system of teleosts, which generally lack highly organized lymphoid tissues. Moreover, the results of this work may be valuable for the development of mucosal vaccines that specifically target M-type cells; mucosal vaccines significantly reduce working costs and the stress that is usually induced by vaccination via injection of individual fish.


Assuntos
Células Apresentadoras de Antígenos/imunologia , Células Epiteliais/imunologia , Doenças dos Peixes/imunologia , Membrana Mucosa/imunologia , Oncorhynchus mykiss/imunologia , Animais , Apresentação do Antígeno/imunologia , Células Apresentadoras de Antígenos/citologia , Células Apresentadoras de Antígenos/metabolismo , Aquicultura/métodos , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/imunologia , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Doenças dos Peixes/microbiologia , Doenças dos Peixes/prevenção & controle , Brânquias/citologia , Brânquias/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Imunidade nas Mucosas , Membrana Mucosa/citologia , Vacinação
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