RESUMO
BACKGROUND: Seasonal allergic rhinitis is a chronic inflammation in the nasal mucosa triggered by inhaled aeroallergens. The inflammatory reaction is controlled by allergen-specific T cells, but where and how these T cells become activated is not fully understood. OBJECTIVES: We wanted to determine whether allergen-specific T-helper (Th) 2 cells are residing in the nasal mucosa under steady-state conditions outside of the pollen season and, if so, whether these cells are activated locally in response to allergen challenge. METHODS: Mucosal biopsies from the lower turbinate were obtained out of season from patients with either birch- or grass-pollen-allergic rhinitis and from healthy controls. Cultured explant samples were challenged with relevant pollen extract or with a mix of overlapping 20-mer peptides derived from the sequence of the major birch allergen, Betula verrucosa (Bet v) 1. After 24 h, culture medium was harvested for multiplex cytokine and tryptase analysis. RESULTS: Significant amounts of interleukin (IL)-5 were secreted from resident cells in response to ex vivo allergen challenge in the allergic group only. No increase was observed for the other cytokines measured. Production of IL-5 in response to both extract and the Bet v1-derived peptide mix strongly suggested that T cells were a major source of IL-5. CONCLUSION: Our explant model indicated that local presentation of antigen to resident allergen-specific Th2 cells is the early event in the pathogenesis of allergic rhinitis. These findings identify possible cellular targets for anti-inflammatory treatment.
Assuntos
Antígenos de Plantas/imunologia , Interleucina-5/imunologia , Modelos Imunológicos , Mucosa Nasal/imunologia , Rinite Alérgica/imunologia , Células Th2/imunologia , Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Mucosa Nasal/patologia , Rinite Alérgica/patologia , Células Th2/patologia , Técnicas de Cultura de TecidosRESUMO
All cases of ichthyosis prematurity syndrome (IPS), registered at the National Center for Fetal Medicine in Trondheim, Norway between 1987 and 2010 were identified and the findings analyzed. Five fetuses with IPS were identified between 1988 and 2000. All five developed polyhydramnios between 28 and 31 weeks. The fetal stomach appeared to be empty in four cases, and was not described in one case. The fetal skin was described as 'uneven' at ultrasound examination in two cases. Separation of chorionic and amniotic membranes with a peculiar appearance of echo-free fluid in the chorionic cavity and echogenic sediment in the amniotic cavity were observed between 28 + 5 and 32 + 3 weeks in all cases. All fetuses were delivered prematurely between 30 and 34 weeks. All neonates had difficulties in breathing, two developed aspiration pneumonia, and one had bilateral pneumothorax after intubation and died at 6 months because of pulmonary and cardiac sequelae. Prenatal sonographic signs of IPS are separation of the membranes, echogenic amniotic fluid and echo-free chorionic fluid occurring between 28 and 32 weeks' gestation. Delivery occurs at 30-34 weeks and, as there is a high risk of asphyxia, an experienced neonatal intensive care unit team should be present at delivery.
Assuntos
Âmnio/diagnóstico por imagem , Córion/diagnóstico por imagem , Ictiose/diagnóstico por imagem , Doenças do Prematuro/diagnóstico por imagem , Ultrassonografia Pré-Natal , Adulto , Líquido Amniótico , Aniridia , Feminino , Humanos , Ictiose/embriologia , Ictiose/mortalidade , Recém-Nascido , Doenças do Prematuro/embriologia , Doenças do Prematuro/mortalidade , Rim/anormalidades , Masculino , Noruega/epidemiologia , Gravidez , Resultado da Gravidez , Transtornos Psicomotores , Ultrassonografia Pré-Natal/métodosRESUMO
BACKGROUND: It has been suggested that Foxp3(+) regulatory T (Treg) cells inhibit allergic inflammation in humans by suppressing the activation of allergen-specific effector T cells. Whether this occurs at the site of allergen exposure has not been determined. OBJECTIVE: To determine the occurrence of Foxp3(+) Treg cells in the nasal mucosa of allergic rhinitis (AR) patients and non-allergic controls after a nasal allergen challenge. METHODS: Pollen-allergic patients (n=18) and non-allergic volunteers (n=7) were challenged locally with pollen extract or placebo for 7 days outside the pollen season. Mucosal biopsies were obtained from the inferior turbinate on days 0, 1 and 7 and subjected to multi-colour immunofluorescence and blood was drawn for eosinophil counts on days 0, 2, 5 and 7. RESULTS: Only AR patients receiving pollen extract experienced typical allergic symptoms and demonstrated increased levels of eosinophils in peripheral blood and nasal mucosa. In allergic patients, a transient early increase (day 1) in CD3(+) T cells was observed in the nasal mucosa, followed by a significant increase of Foxp3(high) T cells at day 7. No changes were found in the control group. The majority of Foxp3(high) cells co-expressed CTLA-4, CD25 and CD4, and a substantial fraction expressed the proliferation marker Ki67. CONCLUSION AND CLINICAL RELEVANCE: Experimentally induced inflammation in AR patients leads to an early inflammatory response followed by accumulation of Foxp3(high) T cells in the nasal mucosa. Our findings are similar to that observed in allergic airways of experimental mice, which suggest that Treg cells are operative in allergic upper airway inflammation. It should be explored whether Treg cells accumulating in the nasal mucosa could be targets for therapeutic intervention.
Assuntos
Fatores de Transcrição Forkhead/metabolismo , Rinite Alérgica Sazonal/imunologia , Linfócitos T Reguladores/imunologia , Adulto , Alérgenos/imunologia , Animais , Gatos , Cães , Eosinófilos/imunologia , Humanos , Masculino , Mucosa Nasal/citologia , Mucosa Nasal/imunologia , Testes de Provocação Nasal , Pólen/imunologia , Rinite Alérgica Sazonal/metabolismo , Rinite Alérgica Sazonal/fisiopatologia , Testes Cutâneos , Linfócitos T Reguladores/metabolismo , Adulto JovemRESUMO
Coeliac disease is a chronic inflammation of the intestinal mucosa controlled by gluten-specific T cells restricted by disease-associated HLA-DQ molecules. We have previously reported that mucosal CD11c(+) dendritic cells (DCs) are responsible for activation of gluten-reactive T cells within the coeliac lesion. In mice, intestinal CD11c(+) DCs comprise several functionally distinct subsets. Here, we report that HLA-DQ(+) antigen-presenting cells (APCs) in normal human duodenal mucosa can be divided into four subsets with striking similarities to those described in mice: CD163(+) CD11c(-) macrophages (74%), and CD11c(+) cells expressing either CD163 (7%), CD103 (11%) or CD1c (13%). CD103(+) and CD1c(+) DCs belonged to partly overlapping populations, whereas CD163(+) CD11c(+) APCs appeared to be a distinct population. In the coeliac lesion, we found increased density of CD163(+) CD11c(+) APCs, whereas the density of CD103(+) and CD1c(+) DCs was decreased, suggesting that distinct subpopulations of APCs in coeliac disease may exert different functions in the pathogenesis.
Assuntos
Antígenos CD/imunologia , Antígenos de Diferenciação Mielomonocítica/imunologia , Antígeno CD11c/imunologia , Doença Celíaca/imunologia , Células Dendríticas/imunologia , Antígenos HLA-DQ/imunologia , Cadeias alfa de Integrinas/imunologia , Receptores de Superfície Celular/imunologia , Adulto , Idoso , Doença Celíaca/patologia , Contagem de Células , Duodeno/imunologia , Duodeno/patologia , Feminino , Humanos , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Macrófagos/imunologia , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
BACKGROUND: Interferon (IFN)-α-producing plasmacytoid dendritic cells (pDCs), inflammatory CD11c+CD1c- myeloid dendritic cells (mDCs) and macrophages have been found to contribute to the pathogenesis of psoriasis. Heliotherapy is a well-established treatment modality of this disease, although the details of how the effects are mediated are unknown. OBJECTIVES: To test the hypothesis that exposure to natural sun affects pathogenic DC subsets in lesional skin. METHODS: Skin biopsies were obtained from lesional and nonlesional skin in 10 patients with moderate to severe psoriasis subjected to controlled sun exposure on Gran Canaria. Biopsies were obtained at baseline, day 2 and day 16 and examined by immunohistochemistry. RESULTS: Sixteen days of heliotherapy had excellent clinical effect on patients with psoriasis, with significant reductions in Psoriasis Area and Severity Index (PASI) scores. In lesional skin pDC numbers and expression of MxA, a surrogate marker for IFN-α, were rapidly reduced. Inflammatory CD11c+CD1c- mDCs were significantly reduced whereas resident dermal CD11c+CD1c+ mDCs were unaffected. Expression levels of the maturation marker DC-LAMP (CD208) on mDCs were significantly reduced after sun exposure, as were the numbers of lesional dermal macrophages. A decrease of dermal DC subsets and macrophages was already observed after 1 day of sun exposure. An additional finding was that DC-SIGN (CD209) is primarily expressed on CD163+ macrophages and not DCs. CONCLUSIONS: The clinical improvement in psoriasis following sun exposure is associated with rapid changes in dermal DC populations and macrophages in lesional skin, preceding the clinical effect. These findings support the concept that these DC subsets are involved in the pathogenesis of psoriasis and suggest that sun-induced clinical benefit may partly be explained by its effect on dermal DCs.
Assuntos
Células Dendríticas/efeitos da radiação , Helioterapia/métodos , Células de Langerhans/efeitos da radiação , Psoríase/patologia , Luz Solar , Adulto , Idoso , Antígenos CD1/metabolismo , Antígenos CD11/metabolismo , Feminino , Proteínas de Ligação ao GTP/metabolismo , Glicoproteínas/metabolismo , Humanos , Imuno-Histoquímica , Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Proteínas de Resistência a Myxovirus , Psoríase/etiologia , Psoríase/terapia , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: Ultraviolet (UV) radiation has immunosuppressive effects and heliotherapy is a well-described treatment modality for psoriasis. OBJECTIVES: To characterize early sun-induced immunological changes both local and systemic in patients with psoriasis. METHODS: Twenty patients with moderate to severe psoriasis were subjected to controlled sun exposure on Gran Canaria, Canary Islands, Spain. Psoriasis Area and Severity Index (PASI) scores were evaluated. Skin biopsies were obtained from lesional and nonlesional skin in 10 patients at baseline and on day 16 and from five additional patients on day 2. Specimens were examined with immunohistochemistry and polymerase chain reaction. Blood samples were obtained from all patients at the same time points and were examined for T-cell subsets and cytokine production. RESULTS: Significant clinical improvement was achieved during the study period. CD4+ and CD8+ T cells in lesional skin were significantly reduced in both the epidermis and dermis. In contrast, dermal FOXP3+ T cells were relatively increased. In the peripheral blood skin homing cutaneous lymphocyte-associated antigen (CLA)+ T cells were significantly decreased after only 1 day in the sun and in vitro stimulated peripheral blood mononuclear cells demonstrated reduced capacity to secrete cytokines after 16 days. CONCLUSIONS: Our data show that clinical improvement of psoriasis following sun exposure is preceded by a rapid reduction in local and systemic inflammatory markers, strongly suggesting that immune modulation mediated the observed clinical effect. We cannot completely rule out that other mechanisms, such as stress reduction, may contribute, but it is extensively documented that UV irradiation is a potent inducer of immunosuppression and we therefore conclude that the observed effect was primarily due to sun exposure.
Assuntos
Citocinas/análise , Helioterapia , Psoríase/imunologia , Psoríase/radioterapia , Pele/imunologia , Pele/efeitos da radiação , Adulto , Idoso , Biópsia , Feminino , Humanos , Imuno-Histoquímica , Células de Langerhans/patologia , Leucócitos Mononucleares/imunologia , Masculino , Pessoa de Meia-Idade , Psoríase/patologia , Índice de Gravidade de Doença , Subpopulações de Linfócitos T/imunologia , Adulto JovemRESUMO
Interleukin (IL)-8, a C-X-C chemokine, activates integrin-mediated adhesion of neutrophils. Presentation of IL-8 on the endothelial cell surface may promote leukocyte extravasation. We found that cultured human microvascular endothelial cells from the intestine (HIMEC) and from nasal polyps (PMEC), but not human umbilical vein endothelial cells (HUVEC), contained IL-8 in intracellular granules that coexpressed von Willebrand factor (vWf ). This observation was corroborated by the immunohistochemical observation of double-positive granules (IL-8(+)vWf+) in vessels of small and large intestine, nasal mucosa, and skin, whereas umbilical cords revealed no endothelial IL-8. After treatment of HIMEC or PMEC with histamine or thrombin, a dramatic increase in supernatant IL-8 concentration was observed within 3 min, whereas no increase in IL-8 was detected in supernatants of identically treated HUVEC cultures. Histamine or thrombin treatment also caused IL-8-containing granules to rapidly disappear from HIMEC. In HUVEC, IL-8-containing granules were inducible by treatment with recombinant human IL-1beta for 24 h; additional histamine treatment doubled IL-8 secretion from HUVEC in the same rapid manner observed for mucosal EC. These data suggested that IL-8 prestored in microvascular endothelial cells may provide a rapid pathway for specific activation of neutrophil adhesion at sites of acute inflammation.
Assuntos
Endotélio Vascular/imunologia , Endotélio Vascular/fisiologia , Interleucina-8/metabolismo , Adesão Celular/fisiologia , Células Cultivadas , Endotélio Vascular/efeitos dos fármacos , Histamina/farmacologia , Humanos , Inflamação/etiologia , Inflamação/imunologia , Microcirculação/efeitos dos fármacos , Microcirculação/imunologia , Microcirculação/fisiologia , Microscopia de Fluorescência , Neutrófilos/fisiologia , Organelas/imunologia , Organelas/metabolismo , Trombina/farmacologia , Veias Umbilicais/efeitos dos fármacos , Veias Umbilicais/imunologia , Veias Umbilicais/fisiologia , Fator de von Willebrand/metabolismoRESUMO
BACKGROUND: In early childhood, the ability to mount protective immune responses in the airways is impaired, with increased risk of allergic sensitisation to inhaled allergens. Antigen presenting cells (APC) and regulatory T cells (Treg) are important modifiers of T cell immunity but little is known about their distribution in bronchial mucosa at this age. Here the subset distribution of APC and the appearance of Foxp3(+) Treg and bronchus associated lymphoid tissue (BALT) were examined immunohistochemically in children less than 2 years of age with chronic asthma-like symptoms of the lower airways. METHODS: Immunophenotyping was performed in situ on bronchial biopsy specimens obtained from 45 infants, 4-23 months of age, under investigation for airway disease. RESULTS: A well developed HLA-DR(+) network of APC was present in all samples, approximately 50% of the cells being CD68(+) macrophages and the remainder various subsets of dendritic cells. The density of HLA-DR(+) cells increased significantly with age but was not related to atopy, clinical symptoms or lung function. Comparing the density of APC subsets and clinical parameters, only the number of intraepithelial CD1a(+) dendritic cells was significantly increased in infants who had recently suffered a respiratory infection. BALT structures were identified in 22 children, with no relation to lung function, atopic status or human rhinovirus positivity. Plasmacytoid dendritic cells and Foxp3(+) Treg were located primarily within these isolated lymphoid follicles. CONCLUSION: A bronchial network of dendritic cells and macrophages develops quite rapidly after birth, apparently independent of clinical symptoms or atopy. The high frequency of BALT structures containing putative tolerogenic dendritic cells and Treg suggests that these lymphoid follicles play an important role in bronchial immune homeostasis during infancy.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Brônquios/imunologia , Linfócitos T Reguladores/imunologia , Antígenos CD/metabolismo , Biomarcadores/metabolismo , Pré-Escolar , Feminino , Fatores de Transcrição Forkhead/metabolismo , Humanos , Imunidade Celular , Imuno-Histoquímica , Lactente , Tecido Linfoide/imunologia , Masculino , Fenótipo , Infecções Respiratórias/imunologiaRESUMO
Regulatory T cells (T(regs)) may inhibit immunity against cancer. Induction and expansion of T(regs) in the immunosuppressive microenvironment created by a growing tumour appear to be one of the mechanisms by which it can evade host defence. We studied the impact of CD25+ T(regs) in a B cell lymphoma model in which Rag2-/- mice received adoptive transfer of wild-type spleen cells with or without CD25+ cells, and concurrently subcutaneous inoculation of the B cell lymphoma cell line A20. We also examined the effect of engaging the glucocorticoid-induced tumour necrosis factor receptor (GITR) - an approach reported previously to abrogate the suppressive effects of T(regs). Mice that received spleen cells depleted of CD25+ T(regs) showed significantly slower tumour growth and increased survival compared with mice that received unsorted spleen cells. The T(reg)-depleted group also had significantly more CD8+ T cells infiltrating the tumours and higher levels of serum immunoglobulin G subclasses. The anti-GITR treatment had no significant effect on tumour growth, survival or immunoglobulin production. In the CD25-depleted group four of 10 mice developed clinical signs of autoimmunity, in contrast to none in the non-depleted group. Forkhead box P3+ T cells were found in tumour-draining lymph nodes in mice in the CD25-depleted group, suggesting an in vivo induction or expansion of rare transferred donor T(regs). Thus, our study showed that removal of CD25+ T(regs) enhanced anti-tumour immunity against local growth of a B cell lymphoma and that induction or expansion of T(regs) could be one mechanism by which the growing tumour evades immune surveillance.
Assuntos
Subunidade alfa de Receptor de Interleucina-2/análise , Depleção Linfocítica , Linfoma de Células B/imunologia , Linfócitos T Reguladores/imunologia , Transferência Adotiva , Animais , Anticorpos Monoclonais/imunologia , Divisão Celular , Modelos Animais de Doenças , Proteína Relacionada a TNFR Induzida por Glucocorticoide , Imunoglobulina G/biossíntese , Imunoglobulina G/sangue , Imunofenotipagem , Linfócitos do Interstício Tumoral/imunologia , Linfoma de Células B/patologia , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Receptores de Fator de Crescimento Neural/imunologia , Receptores do Fator de Necrose Tumoral/imunologia , Baço/imunologiaRESUMO
BACKGROUND: It has been suggested that allergic diseases are caused by defective suppression of allergen-specific Th2 cells by CD4(+)CD25(+) regulatory T cells. However, such studies have been hampered by the difficulty in distinguishing regulatory T cells from CD25-expressing activated T cells. Recently, it was shown that conventional T cells expressed high levels of CD127, whereas regulatory T cells were CD127(lo), allowing discrimination between these distinct T cell subpopulations. OBJECTIVE: The aim of this study was to study whether the putative regulatory subset defined as CD4(+)CD25(+)CD127(lo) was involved in grass pollen-reactive T cell responses. METHODS: Peripheral blood mononuclear cells (PBMCs) were obtained from allergic donors and non-atopic controls out of season. Grass pollen-induced cytokine production and proliferation were compared in cultures of undepleted cells and cells depleted of CD4(+)CD25(+), CD4(+)CD25(+)CD127(hi) or CD4(+)CD25(+)CD127(lo) T cells. RESULTS: Undepleted cell cultures from allergic patients showed significantly increased proliferation and Th2 cytokine production compared with non-atopic controls. Depletion of all CD25(+) T cells did not increase cytokine production or proliferation, and more importantly, no increase in Th2 cytokine production or proliferation was observed in cell cultures depleted of CD4(+)CD25(+)CD127(lo) cells (putative regulatory T cells) compared with undepleted PBMCs in both the allergic and the non-atopic group. CONCLUSION: Our study showed that T cells from grass pollen-allergic patients and non-atopic controls responded very differently to grass pollen extract, but this difference could not be explained by differences in regulatory T cell function. Further studies are needed to understand the importance of regulatory T cells in allergy.
Assuntos
Antígenos de Plantas/imunologia , Subunidade alfa de Receptor de Interleucina-7/análise , Ativação Linfocitária/imunologia , Rinite Alérgica Sazonal/imunologia , Linfócitos T Reguladores/imunologia , Linfócitos T/imunologia , Adulto , Proliferação de Células , Feminino , Humanos , Interferon gama/metabolismo , Interleucina-13/metabolismo , Subunidade alfa de Receptor de Interleucina-2/análise , Interleucina-5/metabolismo , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Teste de Cultura Mista de Linfócitos , Masculino , Pessoa de Meia-Idade , Phleum/imunologia , Pólen/imunologia , Subpopulações de Linfócitos T/química , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Linfócitos T/metabolismo , Linfócitos T Reguladores/química , Linfócitos T Reguladores/metabolismo , Adulto JovemRESUMO
The tissue dendritic cell (DC) compartment is heterogeneous, and the ontogeny and functional specialization of human tissue conventional DC (cDC) subsets and their relationship with monocytes is unresolved. Here we identify monocyte-related CSF1R+Flt3- antigen presenting cells (APCs) that constitute about half of the cells classically defined as SIRPα+ DCs in the steady-state human small intestine. CSF1R+Flt3- APCs express calprotectin and very low levels of CD14, are transcriptionally related to monocyte-derived cells, and accumulate during inflammation. CSF1R+Flt3- APCs show typical macrophage characteristics functionally distinct from their Flt3+ cDC counterparts: under steady-state conditions they excel at antigen uptake, have a lower migratory potential, and are inefficient activators of naïve T cells. These results have important implications for the understanding of the ontogenetic and functional heterogeneity within human tissue DCs and their relation to the monocyte lineage.
Assuntos
Células Dendríticas/fisiologia , Intestinos/fisiologia , Macrófagos/fisiologia , Monócitos/fisiologia , Transcrição Gênica/fisiologia , Transcriptoma/fisiologia , Idoso , Idoso de 80 Anos ou mais , Células Apresentadoras de Antígenos/metabolismo , Células Apresentadoras de Antígenos/fisiologia , Linhagem da Célula/fisiologia , Células Dendríticas/metabolismo , Feminino , Humanos , Inflamação/metabolismo , Inflamação/fisiopatologia , Receptores de Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , Receptores de Fator Estimulador das Colônias de Granulócitos e Macrófagos/metabolismo , Linfócitos T/metabolismo , Linfócitos T/fisiologia , Tirosina Quinase 3 Semelhante a fms/metabolismoRESUMO
Thymic stromal lymphopoietin (TSLP) has multifaceted immunological functions ranging from maintenance of tolerance to induction of disease. Two human transcript variants of TSLP are described: a long form (variant 1; lfTSLP) consisting of four exons and an alternative, short form (variant 2; sfTSLP) that lacks two exons compared with variant 1. SfTSLP has not been described at the protein level or functionally studied. Here, we demonstrate that the human sfTSLP is the predominant form of TSLP, constitutively expressed at the mRNA and protein level in keratinocytes of oral mucosa and skin and in salivary glands, is released in saliva, and is not regulated in the same manner as the long form. Compared with lfTSLP, sfTSLP exhibits a markedly stronger antibacterial activity. Synthetic sfTSLP did not activate signal transducer and activator of transcription 5 (STAT5) signaling in CD1c(+) dendritic cells nor interfered with STAT5 activation by lfTSLP. SfTSLP may, therefore, act as an antimicrobial peptide in the oral cavity and on the skin to create a defense barrier that aids in the control of both commensal and pathogenic microbes. The results show that the two translational products of the TSLP gene have a different expression and different biological properties, and emphasize the importance of analyzing the two TSLP isoforms separately.
Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Bactérias/efeitos dos fármacos , Citocinas/metabolismo , Células Dendríticas/imunologia , Fungos/efeitos dos fármacos , Queratinócitos/imunologia , Peptídeos Catiônicos Antimicrobianos/genética , Peptídeos Catiônicos Antimicrobianos/imunologia , Bactérias/crescimento & desenvolvimento , Células Cultivadas , Citocinas/genética , Citocinas/imunologia , Fungos/crescimento & desenvolvimento , Regulação da Expressão Gênica , Humanos , Imunidade nas Mucosas , Mucosa Bucal/imunologia , Fosforilação/efeitos dos fármacos , Biossíntese de Proteínas , Isoformas de Proteínas/genética , Fator de Transcrição STAT5/metabolismo , Glândulas Salivares/patologia , Transdução de Sinais/efeitos dos fármacos , Pele/patologia , Linfopoietina do Estroma do TimoRESUMO
Lymphocyte recruitment to peripheral tissues is fundamental for immune surveillance and homeostasis, but the chemokines and chemokine receptors responsible for tissue-specific homing of T cells to the upper airway mucosa have not been determined. To address this, we analyzed the chemokines expressed in the normal human nasal mucosa and found that CCL28 is preferentially expressed at a high level on the lumenal face of vascular endothelial cells in the mucosa. Analysis of the cognate chemokine receptors revealed that close to 50% of the CD4(+) T cells in the human nasal mucosa expressed the CCL28 receptor CCR3, whereas CCR3 was hardly detectable on T cells in the small intestine and skin. In the circulation, CCR3(+) T cells comprised a small subset that did not express homing receptors to the intestine or skin. Moreover, depletion of CCR3(+)CD4(+) T cells abrogated the proliferative response of human blood CD4(+) T cells against the opportunistic nasopharyngeal pathogen Haemophilus influenzae, indicating that the CCR3(+)CD4(+) T-cell subset in the circulation contains antigen specificities relevant for the upper airways. Together, these findings indicate that CCL28-CCR3 interactions are involved in the homeostatic trafficking of CD4(+) T cells to the upper airways.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Quimiocinas CC/metabolismo , Endotélio Vascular/imunologia , Haemophilus influenzae/imunologia , Mucosa Nasal/imunologia , Receptores CCR3/metabolismo , Receptores de Retorno de Linfócitos/metabolismo , Adulto , Idoso , Antígenos de Bactérias/imunologia , Movimento Celular , Proliferação de Células , Células Cultivadas , Feminino , Humanos , Ativação Linfocitária , Depleção Linfocítica , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
The monoclonal antibody (mAb) EG2 has been considered to identify activated eosinophils and several immunohistochemical reports of EG2+ eosinophils in various allergic and other inflammatory disorders have suggested an important pathogenic role for such cells. This study showed that cellular EG2 reactivity, both in peripheral blood and mucosal tissue preparations, depends mainly on the method of sample preparation. Nearly 100% of blood eosinophils from normal individuals were strongly EG2+ when prepared by formalin fixation, whereas only a fraction reacted in the unfixed (64%) or acetone-fixed (60%) state. A significantly increased (p < 0.03) number of EG2+ cells were likewise detected in cryo-sections of inflamed nasal mucosa after formalin fixation compared with acetone fixation. Moreover, virtually all eosinophils were EG2+ in cryo-sections of normal jejunal mucosa fixed in periodate-lysine-(0.5%) paraformaldehyde prior to freezing. Conversely, EG2 reacted only weakly, or failed to react, with many eosinophils in cryo-material not subjected to such pre-fixation, in contrast to adjacent non-eosinophilic cells which were brightly stained. Two-colour immunofluorescence consistently revealed overlapping labelling with EG2 and mAb EG1 or a polyclonal antibody to eosinophil cationic protein in sections of formalin-fixed, paraffin-embedded normal gastrointestinal mucosa. Our findings thus showed that EG2 does not provide reliable immunohistochemical discrimination between resting and activated eosinophils. When optimal pre-fixation of tissue specimens was omitted, EG2 reactivity appeared to be caused, at least in part, by leached antigen adsorbed to adjacent non-eosinophilic cells.
Assuntos
Anticorpos Monoclonais/imunologia , Eosinófilos/imunologia , Eosinófilos/metabolismo , Ribonucleases , Adulto , Proteínas Sanguíneas/imunologia , Proteínas Granulares de Eosinófilos , Feminino , Fixadores/química , Imunofluorescência , Secções Congeladas , Técnicas Histológicas , Humanos , Técnicas Imunoenzimáticas , Masculino , Pessoa de Meia-IdadeRESUMO
Several surface receptors are expressed on eosinophils in vitro depending on the state of cellular activation, but immunohistochemical studies of eosinophil-related diseases have mostly focused on the number of infiltrating eosinophils as well as extracellular deposits of eosinophil granule proteins. The present investigation showed that eosinophils display a characteristic granular appearance in cryo-sections and cytospins by differential interference contrast (DIC) imaging. This approach appeared to be more reliable for identification of these cells in situ than immunohistochemical labelling of eosinophil granule proteins. Moreover, combined with immunofluorescence microscopy DIC imaging facilitated three-colour immunofluorescence phenotyping of eosinophils.
Assuntos
Eosinófilos/imunologia , Eosinófilos/ultraestrutura , Imunofluorescência , Microscopia de Interferência/métodos , Contagem de Células , Grânulos Citoplasmáticos/ultraestrutura , Eosinófilos/metabolismo , Estudos de Avaliação como Assunto , Secções Congeladas , Humanos , Enteropatias/patologia , Mucosa Intestinal/citologia , Microtomia , Mucosa Nasal/patologia , Pólipos Nasais/patologia , FenótipoRESUMO
Immunohistochemistry comprises methods used to recognize tissue components as antigens in situ by means of directly or indirectly labeled antibodies, usually (but not always) derived from another species. When applied to cell preparations, the same methods are called immunocytochemistry (Fig. 1), although some authors also use this term for immunostaining of cellular components in tissue sections. Note that compared with immunostaining of vital cells in suspension or cultured monolayers, the sensitivity of immunohistochemical cell-surface staining is considerably reduced because of the decreased amounts of marker antigen represented by the cross-section of the plasma membrane. Also, for certain cellular markers examined in a tissue section, truly peripheral staining may be difficult to distinguish from a rim of cytoplasmic antigen expression. Fig. 1. Immunohistochemistry (A) is performed on tissue sections whereas immunocytochemistry (B) is performed on cells in suspensions, smears, or monolayers. Antibodies do not penetrate the surface membrane of living cells; therefore, unequivocal peripheral staining is obtained only for cells in suspension or vital monolayers. With tissue sections or dried (and fixed) cells, it is difficult to distinguish peripheral from cytoplasmic immunostaining unless the antigen is a distinct surface membrane marker.
RESUMO
The specific defence of airway mucosae depends primarily on secretory immunity. The B cells involved are initially stimulated in organized mucosa-associated lymphoid tissue, apparently including the tonsils and adenoid. From these inductive sites, memory cells migrate to secretory effector sites where they differentiate terminally to immunoglobulin (Ig)-producing plasma cells. Locally produced Ig consists mainly of J chain-containing dimers and larger polymers of IgA (pIgA) that are selectively transported through glandular cells by an epithelial receptor called secretory component or the pIg receptor. IgG can participate in immune exclusion because it reaches the secretions by passive diffusion. However, its proinflammatory properties render IgG antibodies of local immunopathological importance when elimination of penetrating antigens is unsuccessful. T helper (Th) cells activated in this process may by a Th2 cytokine profile promote persistent inflammation with extravasation and priming of eosinophils. This development appears to be part of the late-phase allergic reaction, perhaps initially driven by interleukin-4 (IL-4) released from mast cells that are subjected to IgE-mediated activation, and subsequently also by Th2 cell activation. Eosinophils are potentially tissue-damaging, particularly after priming with IL-5. Various cytokines up-regulate adhesion molecules on endothelial and epithelial cells, thereby enhancing migration of eosinophils into the mucosa, and perhaps in addition causing aberrant immune regulation within the epithelium. Soluble antigens bombarding the epithelial surfaces normally seem to induce several immunosuppressive mechanisms, but mucosal homeostasis appears less patent in the airways than oral tolerance to dietary antigens operating in the gut.
Assuntos
Mucosa Nasal/imunologia , Sistema Respiratório/imunologia , Linfócitos B/imunologia , Antígenos CD4/imunologia , Antígenos CD8/imunologia , Adesão Celular , Citocinas/imunologia , Eosinófilos/imunologia , Homeostase , Humanos , Imunoglobulina A/imunologia , Imunoglobulina G/imunologia , Mastócitos/imunologiaRESUMO
Celiac disease (CD) is a chronic small intestinal inflammation precipitated by gluten ingestion. According to case reports, interferon (IFN)-α administration may induce development of overt CD. Plasmacytoid dendritic cells (PDCs) were thought to be the source of IFN-α and promote a T helper type 1 response leading to lesion formation. Surprisingly and contradicting to earlier findings, PDCs were described as the main antigen-presenting cells (APCs) in human duodenal mucosa and particularly in CD. Here we show that when assessed by flow cytometry and in situ staining, PDCs represent < 1% of APCs in both normal duodenal mucosa and the celiac lesion. Low levels of IFN-α were detected in the celiac lesion assessed by western blot, reverse transcriptase (RT)-PCR, and immunohistochemistry. In four cell populations sorted from the celiac lesion (based on their expression of HLA-DR and CD45), we found that equally low levels of mRNA for IFN-α were distributed among these cell populations. Together, these results suggest that relatively small amount of IFN-α, produced by a variety of cell types, is present in the celiac mucosa. IFN-λ, a type III IFN important in intestinal antiviral defense, was produced mainly by APCs, but its expression was not increased in the celiac lesion.