RESUMO
Members of the transforming growth factor beta (TGF-beta) family of polypeptides have been implicated in morphogenesis and differentiation in numerous tissues, including the lung. In order to further define effects of TGF-beta signaling in lung morphogenesis, a constitutively active form of TGF-beta1 was expressed in respiratory epithelial cells of the fetal mouse lung in vivo. Expression of TGF-beta1 arrested lung morphogenesis in the pseudoglandular stage of development, inhibiting synthesis of differentiation-dependent proteins, SP-B, SP-C, and CCSP, and maintaining embryonic patterns of staining for thyroid transcription factor-1 (TTF-1) and hepatocyte nuclear factor-3beta (HNF-3beta). The pulmonary mesenchyme was thickened and vascular density was increased by TGF-beta1. TGF-beta1 decreased expression of vascular endothelial growth factor-A (VEGF-A) mRNA and protein, and the abundance of Flk-1 mRNA in the lung mesenchyme. Distribution of platelet-endothelial cell adhesion molecule (PECAM)-1, a marker of pulmonary blood vessels, was altered, and ultrastructural studies demonstrated that TGF-beta1 inhibited vascular development in the fetal lung. TGF-beta1 perturbed both epithelial cell differentiation and formation of the pulmonary vasculature, supporting the concept that precise control of signaling via the TGF-beta receptor pathway is critical for normal lung morphogenesis.
Assuntos
Pulmão/irrigação sanguínea , Fator de Crescimento Transformador beta/metabolismo , Animais , Bromodesoxiuridina/metabolismo , Diferenciação Celular , Fatores de Crescimento Endotelial/genética , Células Epiteliais/citologia , Pulmão/citologia , Pulmão/embriologia , Pulmão/metabolismo , Camundongos , Camundongos Transgênicos , Morfogênese , Proteolipídeos/genética , Proteolipídeos/metabolismo , Surfactantes Pulmonares/genética , Surfactantes Pulmonares/metabolismo , RNA Mensageiro , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta1 , Fator A de Crescimento do Endotélio VascularRESUMO
Transgenic mice in which fibroblast growth factor (FGF)-10 was expressed in the lungs of fetal and postnatal mice were generated with a doxycycline-inducible system controlled by surfactant protein (SP) C or Clara cell secretory protein (CCSP) promoter elements. Expression of FGF-10 mRNA in the fetal lung caused adenomatous malformations, perturbed branching morphogenesis, and caused respiratory failure at birth. When expressed after birth, FGF-10 caused multifocal pulmonary tumors. FGF-10-induced tumors were highly differentiated papillary and lepidic pulmonary adenomas. Epithelial cells lining the tumors stained intensely for thyroid transcription factor (TTF)-1 and SP-C but not CCSP, indicating that FGF-10 enhanced differentiation of cells to a peripheral alveolar type II cell phenotype. Withdrawal from doxycycline caused rapid regression of the tumors associated with rapid loss of the differentiation markers TTF-1, SP-B, and proSP-C. FGF-10 disrupted lung morphogenesis and induced multifocal pulmonary tumors in vivo and caused reversible type II cell differentiation of the respiratory epithelium.
Assuntos
Adenoma/induzido quimicamente , Animais Recém-Nascidos/crescimento & desenvolvimento , Feto/fisiologia , Fatores de Crescimento de Fibroblastos/farmacologia , Neoplasias Pulmonares/induzido quimicamente , Pulmão/embriologia , Pulmão/crescimento & desenvolvimento , Uteroglobina , Adenoma/ultraestrutura , Animais , Doxiciclina , Desenvolvimento Embrionário e Fetal/efeitos dos fármacos , Fator 10 de Crescimento de Fibroblastos , Fatores de Crescimento de Fibroblastos/genética , Peptídeos e Proteínas de Sinalização Intercelular , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Neoplasias Pulmonares/ultraestrutura , Camundongos , Camundongos Transgênicos/genética , Proteínas Nucleares/metabolismo , Peptídeos/metabolismo , Precursores de Proteínas/metabolismo , Proteínas/genética , Proteínas/farmacologia , Proteolipídeos/metabolismo , Proteína C Associada a Surfactante Pulmonar , Surfactantes Pulmonares/metabolismo , RNA Mensageiro/metabolismo , Fator Nuclear 1 de Tireoide , Fatores de Transcrição/metabolismoRESUMO
Differentiation of malignant mesothelioma from adenocarcinoma, particularly from a lung primary, remains a difficult diagnostic problem. Surfactant protein B precursor (pro-SP-B) and thyroid transcription factor 1 (ITF-1) are expressed selectively in the normal respiratory epithelium and in adenocarcinomas of the lung. In this study, we evaluated the utility of pro-SP-B and ITF-1 in distinguishing pulmonary adenocarcinomas and malignant mesotheliomas. Immunoreactivity for pro-SP-B and TTF-1 was examined in paraffin sections of 370 primary lung carcinomas (208 adenocarcinomas, 101 squamous cell carcinomas, and 61 large cell carcinomas) and 95 malignant mesotheliomas, using a pro-SP-B antiserum and a monoclonal TTF-1 antibody with a biotin-streptavidin detection system. Immunostaining for pro-SP-B was detected in 57% of adenocarcinomas, and 20% of large cell carcinomas. Immunoreactivity for TTF-1 was shown in 76% of adenocarcinomas and 26% of large cell carcinomas. Malignant mesotheliomas and squamous cell carcinomas did not stain with either antibody. The expression of pro-SP-B and TTF-1 in adenocarcinomas of the lung but not in malignant mesotheliomas shows that pro-SP-B and TTF-1 staining is useful in differentiating these neoplasms.
Assuntos
Adenocarcinoma/metabolismo , Neoplasias Pulmonares/metabolismo , Mesotelioma/metabolismo , Proteínas Nucleares/metabolismo , Precursores de Proteínas/metabolismo , Proteolipídeos/metabolismo , Fatores de Transcrição/metabolismo , Adenocarcinoma/diagnóstico , Carcinoma de Células Grandes/metabolismo , Carcinoma de Células Escamosas/metabolismo , Diagnóstico Diferencial , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/diagnóstico , Mesotelioma/diagnóstico , Fator Nuclear 1 de TireoideRESUMO
We assessed the temporal-spatial distribution of hepatocyte nuclear factor-3beta (HNF-3beta) in developing human lung and other foregut derivatives. Tissue from 31 fetuses (10-40 weeks) and 24 infants with hyaline membrane disease (HMD) or bronchopulmonary dysplasia (BPD) (2 days to 7 months) was studied. HNF-3beta was detected in nuclei of epithelial cells of trachea and of conducting and terminal airways at 10 weeks. Thereafter, epithelial nuclei were immunolabeled more widely in peripheral than proximal airways. HNF-3beta was confined to bronchiolo-alveolar portals and Type II cells in nonfetal lung. In infants with BPD, HNF-3beta was expressed abundantly in regenerating epithelial cells at the periphery of lung lobules. HNF-3beta was also detected in fetal esophagus, pancreas, duodenum, stomach, and gallbladder, suggesting that it is a marker for progenitor cells in foregut derivatives. The pattern of expression of HNF-3beta in the lung was similar to that of thyroid transcription factor-1 (TTF-1) at all ages. The temporal-spatial patterns of HNF-3beta and TTF-1 in the developing and regenerating lung are consistent with their proposed role in epithelial cell differentiation, regeneration, and surfactant protein gene expression. (J Histochem Cytochem 46:955-962, 1998)
Assuntos
Brônquios/metabolismo , Proteínas de Ligação a DNA/metabolismo , Esôfago/metabolismo , Pulmão/metabolismo , Proteínas Nucleares/metabolismo , Traqueia/metabolismo , Fatores de Transcrição/metabolismo , Brônquios/embriologia , Brônquios/crescimento & desenvolvimento , Epitélio/embriologia , Epitélio/crescimento & desenvolvimento , Epitélio/metabolismo , Esôfago/embriologia , Esôfago/crescimento & desenvolvimento , Feto , Fator 3-beta Nuclear de Hepatócito , Humanos , Imuno-Histoquímica , Lactente , Recém-Nascido , Pulmão/embriologia , Pulmão/crescimento & desenvolvimento , Especificidade de Órgãos , Precursores de Proteínas/metabolismo , Proteolipídeos/metabolismo , Proteínas Associadas a Surfactantes Pulmonares , Surfactantes Pulmonares/metabolismo , Regeneração , Fator Nuclear 1 de Tireoide , Traqueia/embriologia , Traqueia/crescimento & desenvolvimentoRESUMO
Antibodies against the pulmonary neuroendocrine cell peptides gastrin-releasing peptide (bombesin), calcitonin and calcitonin gene-related peptide (CGRP) have been labeled with colloidal gold spheres for immunocytochemical localization in human fetal and newborn lung tissue. In general, the presence and amount of immunolabeling increased with increasing gestational age, with only calcitonin appearing late in fetal life. The largest percentage of neuroepithelial body (NEB) cells labeled and the largest number of labeled dense core vesicles (DCV) were in infants with chronic lung disease (bronchopulmonary dysplasia). Serial ribbons allowed identification of more than one peptide in a single NEB cell. The use of two antibodies labeled with colloidal gold spheres of different sizes allowed the identification of two peptides in the same DCV. Quantification of relative amounts of labeled peptides was not possible, as the peptide labeling with the larger size gold sphere was consistently underestimated. Colocalization to the same DCV has been shown in humans for bombesin and calcitonin, calcitonin and CGRP, bombesin and CGRP and, by others for cholecystokinin (CCK) and serotonin. Colocalization of two or more peptides or an amine to a single DCV within the same cell implies simultaneous discharge by exocytosis. The action of the two (or more) substances might be in concert, perhaps with one acting in a paracrine fashion, and the second in an autocrine fashion. In this case, the second peptide or amine might have a regulatory function in the parent cell, influencing DCV storage or rate of release.
Assuntos
Pulmão/citologia , Sistemas Neurossecretores/citologia , Bombesina/análise , Calcitonina/análise , Peptídeo Relacionado com Gene de Calcitonina/análise , Humanos , Imuno-Histoquímica , Recém-Nascido , Pulmão/química , Microscopia Imunoeletrônica , Sistemas Neurossecretores/químicaRESUMO
Surfactant protein B (SP-B) is a 79-amino acid, hydrophobic protein that plays important roles in surfactant function and homeostasis. SP-B is produced in the respiratory epithelium by proteolytic processing of a glycosylated precursor (pro-SP-B) of relative molecular mass 42,000 to 46,000. To develop diagnostic markers for pulmonary adenocarcinomas, we examined the incidence and distribution of pro-SP-B and SP-B mRNA in paraffin sections of 35 non-small cell lung carcinomas (15 adenocarcinomas, 15 squamous cell carcinomas, and 5 large cell carcinomas), using immunohistochemical techniques and in situ hybridization. Fifteen nonpulmonary adenocarcinomas were used as controls. Pro-SP-B and SP-B mRNA were detected in 60% and 53% of pulmonary adenocarcinomas, respectively. Expression was seen in adenocarcinomas with acinar, papillary, bronchioloalveolar, and solid growth patterns. Squamous cell and large cell carcinomas of the lung and nonpulmonary adenocarcinomas did not contain pro-SP-B immunoreactivity or SP-B mRNA. The specificity of SP-B gene expression in adenocarcinomas of the lung supports the usefulness of pro-SP-B and SP-B mRNA in the study and diagnosis of these neoplasms.
Assuntos
Adenocarcinoma/metabolismo , Neoplasias Pulmonares/metabolismo , Precursores de Proteínas/biossíntese , Proteolipídeos/biossíntese , Surfactantes Pulmonares/biossíntese , RNA Mensageiro/biossíntese , Adenocarcinoma/genética , Adenocarcinoma/patologia , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma de Células Escamosas/metabolismo , Humanos , Imuno-Histoquímica , Hibridização In Situ , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Proteolipídeos/genética , Surfactantes Pulmonares/genéticaRESUMO
Clara cell-specific 10-KD protein (CCSP) is an abundant product of nonciliated bronchiolar epithelial (Clara) cells in the lung. We have determined the temporal-spatial distribution of CCSP and its mRNA in developing human lung and in neonatal lung disease, using immunohistochemistry and in situ hybridization. CCSP immunoreactivity was found in nonciliated bronchiolar epithelial cells from 12 weeks of gestation onward. Tracheal and bronchial epithelia showed positive immunoreactivity at each gestational week after 15 weeks and 14 weeks, respectively. CCSP mRNA was seen in the bronchial and bronchiolar epithelia from 16 weeks onward and was detected in the trachea from 19 through 23 weeks of gestation. CCSP immunoreactivity and mRNA were present in nonciliated single cells of bronchial and bronchiolar epithelia in fetuses and in infants with and without lung disease. CCSP- and CCSP mRNA-containing epithelial cells also formed dusters around neuroepithelial bodies (NEBs), especially at airway branch points, suggesting that NEBs and Clara cells might interact during development and during pulmonary regeneration. Because of evidence of overlapping of some but not all cells expressing CCSP, SP-A, and pro-SP-B during lung development, a common cell lineage is proposed, with subsequent divergence of phenotypes.
Assuntos
Displasia Broncopulmonar/metabolismo , Pulmão/metabolismo , Proteínas/metabolismo , RNA Mensageiro/metabolismo , Uteroglobina , Humanos , Doença da Membrana Hialina/metabolismo , Recém-Nascido , Pulmão/citologia , Pulmão/embriologia , Proteínas/genética , Estudos RetrospectivosRESUMO
We assessed the immunohistochemical localization of thyroid transcription factor-1 (TTF-1) in the lungs of 24 human fetuses (11-23 weeks), three infants without pulmonary pathology (36-42 weeks), and 24 infants (2 days-6.5 months) with hyaline membrane disease (HMD) or bronchopulmonary dysplasia (BPD). TTF-1 was detected in fetal lung epithelial cell nuclei by 11 weeks' gestation. Budding tips of terminal airways had prominently labeled nuclei. By 17 weeks, labeling was present in scattered nonciliated columnar and cuboidal cells. Throughout gestation, TTF-1 nuclear staining was prominent in airways abutting pleural, peribronchial, or perivascular connective tissue, being less prominent in centers of lobules. By 23 weeks, many cells in cuboidal but not columnar cell-lined airways had labeled nuclei. At term, TTF-1 was detected primarily in Type II epithelial cells. In HMD with alveolar hemorrhage, edema, or airway collapse, little or no TTF-1 was present except in open terminal airways. In BDP lungs, TTF-1 was absent in areas of alveolar collapse or infection, being present in regenerating open airways. The temporal-spatial distribution of TTF-1, in general, follows patterns of distribution of surfactant protein-B in developing and pathological lungs, consistent with its role in the regulation of epithelial cell gene expression in the lung.
Assuntos
Pulmão/metabolismo , Proteínas Nucleares/biossíntese , Glândula Tireoide/metabolismo , Fatores de Transcrição/biossíntese , Displasia Broncopulmonar/metabolismo , Displasia Broncopulmonar/patologia , Humanos , Doença da Membrana Hialina/metabolismo , Doença da Membrana Hialina/patologia , Técnicas Imunoenzimáticas , Lactente , Recém-Nascido , Pulmão/embriologia , Pulmão/patologia , Glândula Tireoide/patologia , Fator Nuclear 1 de TireoideRESUMO
The human gene encoding thyroid transcription factor-1 (TTF-1), a homeodomain-containing nuclear transcription protein of the Nkx2 gene family, was isolated and characterized. Human TTF-1 was encoded by a single gene locus spanning approximately 3.3 kilobases and consisted of two exons and a single intron. The TTF-1 cDNA and polypeptide of 371 amino acids have been highly conserved, sharing 98% identity with the rat TTF-1 polypeptide. Human TTF-1 mRNA and polypeptide were selectively expressed in human and mouse pulmonary adenocarcinoma cell lines. In addition to its presence in thyroid gland epithelium, the human TTF-1 protein was detected by immunohistochemistry in human fetal lung as early as 11 weeks of gestation, being localized in the nuclei of epithelial cells of the developing airways. After birth, TTF-1 was selectively expressed in Type II epithelial cells in the alveoli and in subsets of bronchiolar epithelial cells in the conducting regions of the lung. The 5'-flanking region of the human TTF-1 gene directed transcription of luciferase cDNA in a lung epithelial cell-selective manner. The conservation and distribution of TTF-1 in the human respiratory tract support its role in the regulation of lung development and surfactant homeostasis.
Assuntos
Pulmão/metabolismo , Proteínas Nucleares/genética , Fatores de Transcrição/genética , Adulto , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA Complementar , Células Epiteliais , Epitélio/metabolismo , Expressão Gênica , Humanos , Imuno-Histoquímica , Recém-Nascido , Pulmão/citologia , Pulmão/embriologia , Dados de Sequência Molecular , Proteínas Nucleares/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator Nuclear 1 de Tireoide , Fatores de Transcrição/biossíntese , Transcrição GênicaRESUMO
The distribution of immunoreactive surfactant-associated protein B (IR-SP-B) was studied immunohistochemically in 120 subjects from 10 weeks of gestation to 7 postnatal months with a polyclonal antibody against human SP-B. Electron microscopy (EM) was done in 72 subjects to document the presence of Type II cells containing lamellar bodies. Fetuses of less than 18 weeks' gestation showed no immunostaining. Beginning at 18 weeks, non-mucous cells of tracheal glands immunostained in a few instances. Fetuses of 19 through 23 weeks showed progressive immunostaining of cells lining terminal airways. Infants 26-40 weeks who died with or without pulmonary pathology showed immunostaining of Type II cells and bronchioloalveolar (BA) portal cells of the respiratory bronchioles. In infants with hyaline membrane disease (HMD) who died less than 12 days after birth, occasional tracheal gland cells, BA portal cells, and mature and relining Type II cells immunostained. In bronchopulmonary dysplasia (BPD), BA portal cells, relining Type II cells, macrophages, and luminal material immunostained. Occasional tracheal and bronchial gland cells and Clara cells immunostained. The appearance of IR-SP-B at mid-gestation correlated with differentiation of Type II cells. There was good correlation of immunostaining with the presence of lamellar bodies on EM. Accelerated maturation of the lung was often associated with premature rupture of membranes (PROM).
Assuntos
Feto/metabolismo , Proteolipídeos/metabolismo , Surfactantes Pulmonares/metabolismo , Humanos , Imuno-Histoquímica , Lactente , Recém-Nascido , Pulmão/anatomia & histologia , Pulmão/metabolismo , Pulmão/patologia , Pneumopatias/metabolismo , Pneumopatias/patologia , Estudos Retrospectivos , Traqueia/anatomia & histologia , Traqueia/metabolismoRESUMO
A model of pulmonary adenocarcinomas was produced in transgenic mice harboring a chimeric gene comprising the SV40 large T antigen under the control of a transcriptional region derived from the human surfactant protein C (SP-C) gene. Transgenic mice succumbed with pulmonary tumors within 4-5 months of age. By histology, the tumors were adenocarcinomas with lepidic, papillary, and solid growth patterns that were indistinguishable from adenocarcinomas occurring in humans. Immunocytochemistry demonstrated the lack of staining for neuroendocrine markers, consistent with the identification of the tumors as non-small cell rather than small cell carcinomas. The presence of SV40 large T mRNA in the lung and tumors was detected by in situ hybridization and Northern blot analysis. Exogenous SV40 large T mRNA and endogenous CC10 (a nonciliated respiratory epithelial cell marker) and SP-C (a Type II alveolar cell marker) mRNAs were expressed at variable levels in the lung tumors. SV40 large T mRNA and CC10 mRNA were detected in the majority of tumors, while SP-C mRNA was detected less frequently. The heterogeneity of bronchiolar and alveolar cell markers in the tumors from the transgenic mice supports the concept that tumorigenesis was initiated in distinct subsets of epithelial cells that produce characteristic adenocarcinomas of the lung.
Assuntos
Adenocarcinoma/genética , Antígenos Transformantes de Poliomavirus/genética , Genes Reguladores/genética , Neoplasias Pulmonares/genética , Proteolipídeos/genética , Surfactantes Pulmonares/genética , Adenocarcinoma/imunologia , Animais , Biomarcadores Tumorais/análise , Northern Blotting , Quimera/genética , Modelos Animais de Doenças , Epitélio/fisiologia , Expressão Gênica/genética , Humanos , Pulmão/fisiologia , Neoplasias Pulmonares/imunologia , Camundongos , Camundongos Transgênicos , Hibridização de Ácido Nucleico , RNA Mensageiro/genéticaRESUMO
We evaluated the distribution of lipocortin-1 immunoreactivity in 118 immature or mature human hypophyses using the peroxidase-antiperoxidase (PAP) technique with a polyclonal rabbit antiserum against lipocortin-1. Serial sections were evaluated for five pituitary hormones and S-100 protein immunoreactivity to compare their distributions with that of lipocortin-1. Scattered or moderate numbers of cells exhibited lipocortin-1 immunoreactivity in the pars distalis of 89 subjects ranging in age from 27 weeks' gestation to 83 years. Seven immature and seven aged specimens exhibited no immunostaining, while 15 specimens from older individuals exhibited only rare immunostaining. Immunostaining did not appear to co-localize selectively with any specific pituitary hormone, although the distribution of immunoreactivity did overlap that of some corticotrophs and was seen in elongated processes of S-100-containing folliculostellate cells. Lipocortin-1 was also found in epithelial cells lining colloid cysts of the residual pars intermedia in 115 of 118 pituitaries ranging in age from 23 weeks' gestation to 83 years. In many intermediate lobe cysts, lipocortin-1 exhibited a pattern of immunoreactivity that partially overlapped the distribution of S-100 protein immunostaining, although the pattern was not identical. Pre-absorption of anti-lipocortin-1 antiserum with lipocortin-1-coupled Sepharose-4B immunoreactivity resulted in loss of immunoreactivity in both lobes. No lipocortin-1 immunoreactivity was seen in the neurohypophysis.
Assuntos
Proteínas de Ligação ao Cálcio/análise , Hipófise/química , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/metabolismo , Anexinas , Criança , Pré-Escolar , Humanos , Lactente , Pessoa de Meia-Idade , Hipófise/embriologia , Adeno-Hipófise/química , Adeno-Hipófise/embriologia , Neuro-Hipófise/química , Neuro-Hipófise/embriologia , Hormônios Hipofisários/análise , Proteínas S100/análise , Distribuição TecidualRESUMO
We hypothesized that changes in plasma retinol-binding protein (RBP) concentration in response to vitamin A administration might be useful for evaluating vitamin A status of very low birth weight infants susceptible to bronchopulmonary dysplasia. We prospectively studied 24 consecutively admitted neonates (birth weight less than 1350 gm, gestational age less than 31 weeks, ventilator dependent for greater than 24 hours after birth), who were eligible to receive 2000 IU supplemental vitamin A by intramuscular injection on postnatal day 1 and on alternate days thereafter for 28 days. In addition to serial assessment of vitamin A status, we measured plasma RBP just before and 1, 3, and 6 hours after an intramuscular injection of vitamin A (2000 IU/kg retinyl palmitate) on days 1 and 28. The percent increase in plasma RBP (delta-RBP) was high (mean +/- SD: 61 +/- 37%) and plasma vitamin A and RBP values were low on day 1, indicative of vitamin A deficiency. Supplemental vitamin A improved vitamin A status of all infants as shown by low delta-RBP (mean +/- SD: 8 +/- 9%) and normal plasma vitamin A and RBP values on day 28. Bronchopulmonary dysplasia was diagnosed in 12 of 24 infants. Infants with bronchopulmonary dysplasia had a higher mean (+/- SD) delta-RBP on day 28 than those without bronchopulmonary dysplasia (13 +/- 10% vs 3 +/- 3%, p less than 0.01), indicative of persistence of low vitamin A status in infants with lung disease despite supplementation. We conclude that the plasma RBP response to vitamin A is a useful indicator of vitamin A status in very low birth weight infants. Although vitamin A supplementation at the dosage used in this study normalizes conventional plasma indexes of vitamin A in very low birth weight infants, the plasma RBP response to vitamin A may continue to reflect persistence of low vitamin A status in the more immature infants with significant lung disease. We suggest that the plasma RBP response to vitamin A may be a useful functional test in such infants.
Assuntos
Displasia Broncopulmonar/sangue , Recém-Nascido de Baixo Peso/sangue , Recém-Nascido Prematuro/sangue , Proteínas de Ligação ao Retinol/análise , Vitamina A/farmacologia , Suscetibilidade a Doenças , Nutrição Enteral , Feminino , Humanos , Fenômenos Fisiológicos da Nutrição do Lactente , Recém-Nascido , Injeções Intramusculares , Masculino , Prognóstico , Estudos Prospectivos , Proteínas Plasmáticas de Ligação ao Retinol , Fatores de Tempo , Vitamina A/administração & dosagem , Vitamina A/sangueRESUMO
The ontogeny and distribution of the epidermal growth factor (EGF) receptor and lipocortin-1, a major cellular substrate of the EGF receptor, were evaluated in a developmental series of fetal and neonatal human lungs (8 to 41 weeks' gestation and stillborn to 16 days' postnatal age). The peroxidase anti-peroxidase technique with two polyclonal antibodies recognizing the EGF receptor and one polyclonal antibody recognizing lipocortin-1 were used for immunohistochemical localization. Extensive or scattered bronchiolar EGF receptor immunoreactivity appeared in the entire series of frozen lung specimens from 15 to 32 weeks' gestation. Bronchial glands exhibited EGF receptor immunostaining from 19 weeks onward, and immunoreactivity in bronchial epithelium was detected from 23 weeks onward. Most tracheas showed extensive lipocortin-1 immunoreactivity in the epithelium beginning at 10 weeks' gestation. Immunostaining was also seen in cells lining the ducts of submucosal glands after 15 weeks' gestation and in nonmucous acinar cells of tracheal glands after their appearance at 18 weeks' gestation. Bronchial epithelium exhibited lipocortin-1 immunoreactivity from 12 weeks' gestation onward. Bronchial gland necks became immunostained from 16 weeks' gestation onward, followed by acinar immunostaining as they subsequently developed. Bronchiolar epithelium was immunostained as early as 12 weeks, beginning with the largest airways, and by 24 weeks extending distally to the bronchioloalveolar portals. Lipocortin-1 immunostaining of larger conducting airway epithelium was primarily confined to ciliated cells. Neither EGF receptor nor lipocortin-1 immunoreactivity was detected in alveolar type I or type II cells, fibrocytes, chondrocytes, or smooth muscle cells at any gestational age. These developmental patterns suggest that the EGF receptor and lipocortin-1 may participate in normal growth factor-induced proliferation of the conducting airways and their glands in the human fetal lung and trachea.
Assuntos
Proteínas de Ligação ao Cálcio/análise , Receptores ErbB/análise , Feto/análise , Pulmão/análise , Fosfolipases/antagonistas & inibidores , Anexinas , Feminino , Feto/citologia , Feto/patologia , Humanos , Imuno-Histoquímica , Recém-Nascido , Pulmão/citologia , Pulmão/patologia , Gravidez , Traqueia/análise , Traqueia/citologia , Traqueia/patologiaRESUMO
We have examined the ontogeny and distribution of the epidermal growth factor receptor/kinase (EGF receptor) and of lipocortin-1, a major cellular substrate of the EGF receptor, in a developmental series of 13 normal ovine fetal lungs (44-145 d of gestation) using the peroxidase anti-peroxidase technique with two extensively characterized polyclonal antibodies recognizing the EFG receptor and one polyclonal antibody recognizing lipocortin-1. Immunoreactive EGF receptor/kinase and lipocortin-1 were detected in conducting airway epithelium by the end of the first trimester of pregnancy before bronchial glands could be identified. This was followed at two-thirds of gestation by immunoreactivity in bronchial glands and large bronchioles adjacent to positive bronchi. By seven-eighths of gestation conducting airway epithelium no longer contained consistently detectable immunostaining for EGF receptor, although lipocortin-1 was identified until term in all levels of conducting airways. In contrast, neither EGF receptor nor lipocortin-1 immunoreactivity was detected in alveolar type I or type II epithelial cells, fibrocytes, chondrocytes, smooth muscle, or endothelial cells at any gestational age. These findings suggest that EGF receptor and lipocortin-1 may participate in early airway development.
Assuntos
Receptores ErbB/metabolismo , Maturidade dos Órgãos Fetais , Glicoproteínas/metabolismo , Pulmão/embriologia , Animais , Anexinas , Epitélio/anatomia & histologia , Epitélio/metabolismo , Idade Gestacional , Técnicas Imunoenzimáticas , Pulmão/anatomia & histologia , Pulmão/metabolismo , OvinosRESUMO
Cells staining for immunoreactive human epidermal growth factor were sought in the lungs and tracheas of human fetuses from 8 to 24 weeks of gestation. Lungs of liveborn infants from 25 to 40 weeks of gestation (stillborn to 7 months postnatal life), both with and without lung pathology, were also studied. In the early fetal trachea (12 to 15 weeks), many nonciliated cells immunostained for immunoreactive human epidermal growth factor in the lining epithelium. By 16 weeks of gestation this widespread staining was replaced by stained nonciliated single cells or small clusters of cells which were identifiable until 24 weeks. In the few tracheas which were available from liveborn infants who died without evidence of lung disease, stained cells were seldom identified in the lining epithelium after 24 weeks of gestation. In contrast, from 18 weeks until term, tracheal submucosal glands contained scattered cells which immunostained for immunoreactive human epidermal growth factor but which did not appear to be classical mucous cells. Beginning at 20 weeks of gestation, positively staining cells were found occasionally in bronchial lining epithelium, but more often in bronchial submucosal glands. Immunostained cells were never identified in bronchiolar epithelium in normal fetal or newborn lungs. In liveborn infants from 24 weeks onward who developed lung disease, many tracheas were severely damaged. In the presence of extensive denudation of the mucosa or the development of squamous metaplasia, immunostained cells were rarely seen in the lining epithelium. However, even under these conditions stained glandular cells could usually be identified. Stained cells were also present in the necks of those tracheal glands from which new epithelial lining cells appeared to be migrating onto denuded surfaces. Immunostained cells in the bronchial lining epithelium of infants with chronic lung disease were infrequent, just as they were in the fetus, but bronchial submucosal glands contained positively stained cells similar to those in tracheal glands. The appearance and distribution of immunostained cells were similar in the tracheal and bronchial submucosal glands in both normal subjects and those with all stages of lung disease. In contrast to the bronchioles of fetuses and infants without lung pathology, the bronchiolar epithelium of infants with chronic lung disease contained immunostained cells. Immunostained cells were found in areas of migrating dysplastic cells in relining conducting airways.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Desenvolvimento Embrionário e Fetal , Fator de Crescimento Epidérmico/análise , Pneumopatias/metabolismo , Sistema Respiratório/análise , Doença Aguda , Displasia Broncopulmonar/metabolismo , Doença Crônica , Feminino , Humanos , Doença da Membrana Hialina/metabolismo , Recém-Nascido , Pneumopatias/fisiopatologia , Masculino , Pessoa de Meia-Idade , Sistema Respiratório/embriologia , Sistema Respiratório/crescimento & desenvolvimento , Estudos RetrospectivosRESUMO
Twelve pairs of fetal lambs were used to test the hypothesis that the necrotizing tracheobronchitis followed by squamous metaplasia seen in premature infants who develop chronic bronchopulmonary dysplasia might be related to low retinol stores and might, therefore, be reversed by retinol supplementation. Epidermal growth factor (EGF) was used to model the growth factor stimulus initiated by chronic wounding of the airways, and retinol was used as a differentiator of proliferating cells stimulated by EGF. Saline-treated animals were used as controls, as were fetal lambs receiving retinol alone or EGF alone. The effects of EGF on tracheal and bronchial epithelium consisted of proliferation of basal and intermediate cells, necrosis and slough of lining ciliated and mucous-producing cells, followed by squamous metaplasia. In fetal lambs given retinol, plasma, liver and lung retinol levels rose and mucous producing cells were increased in number. In the presence of EGF plus retinol, differentiation of mucous-producing cells was accelerated. We believe that this fetal lamb model with low initial levels of retinol in plasma, liver and lung, treated with EGF may mimic human premature infants with chronic bronchopulmonary dysplasia, and that the addition of retinol in amounts sufficient to raise their tissue levels produces a more normal surface epithelium in conducting airways.
Assuntos
Displasia Broncopulmonar/etiologia , Fator de Crescimento Epidérmico/farmacologia , Traqueia/efeitos dos fármacos , Deficiência de Vitamina A/complicações , Vitamina A/farmacologia , Análise de Variância , Animais , Animais Recém-Nascidos , Displasia Broncopulmonar/tratamento farmacológico , Displasia Broncopulmonar/patologia , Diferenciação Celular , Divisão Celular , Modelos Animais de Doenças , Fator de Crescimento Epidérmico/administração & dosagem , Fator de Crescimento Epidérmico/uso terapêutico , Epitélio/efeitos dos fármacos , Epitélio/patologia , Epitélio/ultraestrutura , Feminino , Feto , Humanos , Recém-Nascido , Metaplasia , Microscopia Eletrônica , Gravidez , Ovinos , Traqueia/patologia , Traqueia/ultraestrutura , Vitamina A/administração & dosagem , Vitamina A/uso terapêuticoRESUMO
The ontogeny of calcitonin gene-related peptide immunoreactivity (CGRP-IR) was evaluated immunohistochemically in 67 human fetal or newborn lungs previously analyzed for calcitonin immunoreactivity (CT-IR). CGRP-IR was present by 10 weeks of gestation in rare, solitary neuroendocrine (NE) cells of developing conducting airways in two of eight first-trimester lungs. During the second trimester, cells with CGRP-IR were found consistently (21/23 fetuses). However, the numbers of positively staining cells did not appear to increase in these fetuses or in third-trimester infants dying of non-pulmonary causes. The highest concentrations of CGRP-IR cells were seen in lungs of premature infants with advancing chronic lung disease associated with bronchopulmonary dysplasia (BPD). CGRP-IR was seen earlier in gestation and in greater numbers of NE cells than was calcitonin immunoreactivity (CT-IR) reported previously in these same fetal lungs (Lab Invest 52:52, 1985). Its presence paralleled that of CT-IR in postnatal chronic lung disease.