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1.
Dev Comp Immunol ; 95: 89-95, 2019 06.
Article in English | MEDLINE | ID: mdl-30753854

ABSTRACT

The chicken yolk sac (YS) plays an important role in nutrient absorption and immune function for the developing embryo. The avian ß-defensins (AvBD) are cationic peptides that are important members of the innate immune system. The objective of this study was to profile AvBD mRNA expression patterns and distribution of cells expressing AvBD mRNA in the chicken YS. Expression of AvBD1, 2, 7, and 10 mRNA was low at embryonic day 7 (e7), increased to e9 through e13 and then declined to e19. Using in situ hybridization, AvBD10 mRNA was found to be expressed in endodermal epithelial cells, while AvBD1, 2, and 7 mRNA were expressed in heterophils. The developmental expression pattern and distribution of AvBD mRNA in the YS reveals the importance of these genes to protection of the developing chick embryo.


Subject(s)
Avian Proteins/genetics , Embryonic Development/immunology , Gene Expression Regulation, Developmental/immunology , Yolk Sac/immunology , beta-Defensins/genetics , Animals , Avian Proteins/immunology , Chick Embryo , Chickens , Endoderm/cytology , Endoderm/immunology , Endoderm/metabolism , Epithelial Cells/immunology , Epithelial Cells/metabolism , RNA, Messenger/metabolism , Yolk Sac/growth & development , Yolk Sac/metabolism , beta-Defensins/immunology
2.
Cell Stem Cell ; 16(2): 148-57, 2015 Feb 05.
Article in English | MEDLINE | ID: mdl-25533131

ABSTRACT

Type 1 diabetes (T1D) is an autoimmune disease caused by T cell-mediated destruction of insulin-producing ß cells in the islets of Langerhans. In most cases, reversal of disease would require strategies combining islet cell replacement with immunotherapy that are currently available only for the most severely affected patients. Here, we demonstrate that immunotherapies that target T cell costimulatory pathways block the rejection of xenogeneic human embryonic-stem-cell-derived pancreatic endoderm (hESC-PE) in mice. The therapy allowed for long-term development of hESC-PE into islet-like structures capable of producing human insulin and maintaining normoglycemia. Moreover, short-term costimulation blockade led to robust immune tolerance that could be transferred independently of regulatory T cells. Importantly, costimulation blockade prevented the rejection of allogeneic hESC-PE by human PBMCs in a humanized model in vivo. These results support the clinical development of hESC-derived therapy, combined with tolerogenic treatments, as a sustainable alternative strategy for patients with T1D.


Subject(s)
Diabetes Mellitus, Type 1/therapy , Endoderm/cytology , Endoderm/immunology , Human Embryonic Stem Cells/transplantation , Immune Tolerance/immunology , Pancreas/cytology , T-Lymphocytes, Regulatory/immunology , Animals , Diabetes Mellitus, Type 1/pathology , Humans , Mice , Mice, Inbred NOD , Mice, Knockout , Mice, SCID , Pancreas/immunology , Transplantation, Heterologous
3.
PLoS One ; 8(7): e70863, 2013.
Article in English | MEDLINE | ID: mdl-23923029

ABSTRACT

FcRn, a non-classical MHCI molecule, transports IgG from mother to young and regulates the rate of IgG degradation throughout life. Brambell proposed a mechanism that unified these two functions, saying that IgG was pinocytosed nonspecifically by the cell into an FcRn-expressing endosome, where, at low pH, it bound to FcRn and was exocytosed. This theory was immediately challenged by claims that FcRn specificity for ligand could be conferred at the cell surface in neonatal jejunum. Assessing Brambell's hypothesis we found abundant nonspecifically endocytosed IgG present in the cytoplasm of FcRn(-/-) enterocytes. Further, IgG was present in the intercellular clefts and the cores of FcRn(+/+) but not FcRn(-/-) jejunum. FcRn specificity for ligand could be determined within the cell.


Subject(s)
Endoderm/immunology , Endoderm/metabolism , Enterocytes/immunology , Enterocytes/metabolism , Histocompatibility Antigens Class I/genetics , Immunoglobulin G/immunology , Receptors, Fc/genetics , Animals , Animals, Newborn , Female , Gene Expression , Histocompatibility Antigens Class I/metabolism , Immunoglobulin G/metabolism , Intestinal Mucosa/metabolism , Intestines/immunology , Intracellular Space/metabolism , Jejunum/immunology , Jejunum/metabolism , Mice , Mice, Knockout , Protein Transport , Receptors, Fc/deficiency , Receptors, Fc/metabolism
4.
Stem Cells ; 30(9): 1999-2009, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22761039

ABSTRACT

Monoclonal antibodies against cell surface markers are powerful tools in the study of tissue regeneration, repair, and neoplasia, but there is a paucity of specific reagents to identify stem and progenitor cells in tissues of endodermal origin. The epitope defined by the GCTM-5 monoclonal antibody is a putative marker of hepatic progenitors. We sought to analyze further the distribution of the GCTM-5 antigen in normal tissues and disease states and to characterize the antigen biochemically. The GCTM-5 epitope was specifically expressed on tissues derived from the definitive endoderm, in particular the fetal gut, liver, and pancreas. Antibody reactivity was detected in subpopulations of normal adult biliary and pancreatic duct cells, and GCTM-5-positive cells isolated from the nonparenchymal fraction of adult liver expressed markers of progenitor cells. The GCTM-5-positive cell populations in liver and pancreas expanded greatly in numbers in disease states such as biliary atresia, cirrhosis, and pancreatitis. Neoplasms arising in these tissues also expressed the GCTM-5 antigen, with pancreatic adenocarcinoma in particular showing strong and consistent reactivity. The GCTM-5 epitope was also strongly displayed on cells undergoing intestinal metaplasia in Barrett's esophagus, a precursor to esophageal carcinoma. Biochemical, mass spectrometry, and immunochemical studies revealed that the GCTM-5 epitope is associated with the mucin-like glycoprotein FCGBP. The GCTM-5 epitope on the mucin-like glycoprotein FCGBP is a cell surface marker for the study of normal differentiation lineages, regeneration, and disease progression in tissues of endodermal origin.


Subject(s)
Cell Adhesion Molecules/immunology , Epitopes/biosynthesis , Glycoproteins/immunology , Liver/cytology , Stem Cells/immunology , Cell Differentiation/immunology , Endoderm/cytology , Endoderm/immunology , Epitopes/immunology , Humans , Liver/immunology , Stem Cells/cytology
5.
Stem Cells ; 27(9): 2103-13, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19522011

ABSTRACT

The development of functional cell populations such as hepatocytes and pancreatic beta cells from embryonic stem cell (ESC) is dependent on the efficient induction of definitive endoderm early in the differentiation process. To monitor definitive endoderm formation in mouse ESC differentiation cultures in a quantitative fashion, we generated a reporter cell line that expresses human CD25 from the Foxa3 locus and human CD4 from the Foxa2 locus. Induction of these reporter ESCs with high concentrations of activin A led to the development of a CD25-Foxa3+CD4-Foxa2+ population within 4-5 days of culture. Isolation and characterization of this population showed that it consists predominantly of definitive endoderm that is able to undergo hepatic specification under the appropriate conditions. To develop reagents that can be used for studies on endoderm development from unmanipulated ESCs, from induced pluripotent stem cells, and from the mouse embryo, we generated monoclonal antibodies against the CD25-Foxa3+CD4-Foxa2+ population. With this approach, we identified two antibodies that react specifically with endoderm from ESC cultures and from the early embryo. The specificity of these antibodies enables one to quantitatively monitor endoderm development in ESC differentiation cultures, to study endoderm formation in the embryo, and to isolate pure populations of culture- or embryo-derived endodermal cells.


Subject(s)
Antibodies, Monoclonal/immunology , CD4 Antigens/metabolism , Endoderm/immunology , Gene Expression Regulation, Developmental , Hepatocyte Nuclear Factor 3-beta/metabolism , Hepatocyte Nuclear Factor 3-gamma/metabolism , Interleukin-2 Receptor alpha Subunit/metabolism , Activins/pharmacology , Animals , Cell Differentiation/drug effects , Cell Differentiation/genetics , Embryonic Stem Cells/cytology , Embryonic Stem Cells/immunology , Embryonic Stem Cells/metabolism , Endoderm/cytology , Flow Cytometry , Humans , Mice
6.
Immunobiology ; 214(3): 165-78, 2009.
Article in English | MEDLINE | ID: mdl-19195737

ABSTRACT

The origin of the complement system, one of the major systems of mammalian innate immunity, is more ancient than that of the adaptive immune system, as shown by the identification of the gene for the complement component 3 (C3) in a basic metazoa, cnidarian coral. Only a few reports on the other complement genes of non-chordates have been published, and the composition of the ancient complement system has not been clarified. We performed comprehensive cloning of the complement genes with characteristic domain structures using a Cnidarian, the sea anemone, Nematostella vectensis. Partial sequences of the two C3, two factor B (Bf), and one mannan-binding protein-associated serine protease (MASP) genes were identified in the draft genome data, and the complete coding sequences of these genes were elucidated by RT-PCR and 5'- and 3'-RACE. In contrast, no C6 and factor I family genes were identified. These cnidarian components shared the unique domain structures and most of the functionally critical amino acid residues with their mammalian counterparts, suggesting the conservation of their basic biochemical functions throughout metazoan evolution. In situ hybridization analysis indicated that all five genes are expressed in the tentacles, pharynx, and mesentery in an endoderm-specific manner. These results suggest that the multi-component complement system comprising at least C3, Bf, and MASP was established in a common ancestor of Cnidaria and Bilateria more than 600 million years ago to protect the coelenteron, the primitive gut cavity with putative circulatory functions.


Subject(s)
Complement C3/metabolism , Complement Factor B/metabolism , Complement System Proteins/metabolism , Endoderm/immunology , Mannose-Binding Protein-Associated Serine Proteases/metabolism , Sea Anemones/immunology , Animals , Cloning, Molecular , Complement C3/genetics , Complement C3/immunology , Complement Factor B/genetics , Complement Factor B/immunology , Complement System Proteins/genetics , Complement System Proteins/immunology , Evolution, Molecular , Gene Expression Profiling , Humans , Mannose-Binding Protein-Associated Serine Proteases/genetics , Mannose-Binding Protein-Associated Serine Proteases/immunology , Organ Specificity , Phylogeny , Protein Structure, Tertiary , Sequence Analysis, DNA
7.
J Immunol ; 182(5): 2583-9, 2009 Mar 01.
Article in English | MEDLINE | ID: mdl-19234152

ABSTRACT

In adults, the nonclassical MHC class I molecule, FcRn, binds both IgG and albumin and rescues both from a degradative fate, endowing both proteins with high plasma concentrations. FcRn also transports IgG from mother to young during gestation. Anticipating that a detailed understanding of gestational IgG transport in the mouse may give us a useful model to understand FcRn function in the human placenta, we have studied FcRn in the mouse yolk sac placenta in detail. Analyzing day 19-20 fetuses of the three FcRn genotypes resulting from matings of FcRn(+/-) parents, we found that FcRn(-/-) fetuses showed negligible IgG concentrations (1.5 microg/ml), whereas IgG concentrations in FcRn(+/-) fetuses were about a half (176 microg/ml) that of FcRn(+/+) fetuses (336 microg/ml), indicating that FcRn is responsible for virtually all IgG transport from mother to fetus. Immunofluorescence and immunoblotting studies indicated that FcRn is expressed in the endoderm of the yolk sac placenta but not in other cells of the yolk sac placenta or in the chorioallantoic placenta. IgG was found in the endoderm of both FcRn(+/+) and FcRn(-/-) yolk sac placentas and in the mesenchyme of FcRn(+/+) but was missing from the mesenchyme of FcRn(-/-) yolk sac placentas, indicating that IgG enters the endoderm constitutively but is moved out of the endoderm by FcRn. The similarities of these results to human placental FcRn expression and function are striking.


Subject(s)
Endoderm/immunology , Histocompatibility Antigens Class I/physiology , Immunoglobulin G/metabolism , Receptors, Fc/deficiency , Receptors, Fc/physiology , Yolk Sac/immunology , Animals , Endoderm/metabolism , Female , Fetus/blood supply , Histocompatibility Antigens Class I/biosynthesis , Histocompatibility Antigens Class I/metabolism , Humans , Immunoglobulin G/blood , Male , Maternal-Fetal Exchange/immunology , Mice , Mice, Inbred C57BL , Mice, Knockout , Mice, Transgenic , Placenta/immunology , Placenta/metabolism , Pregnancy , Protein Transport/immunology , Receptors, Fc/biosynthesis , Receptors, Fc/metabolism , Yolk Sac/metabolism
8.
J Immunol ; 182(5): 2997-3007, 2009 Mar 01.
Article in English | MEDLINE | ID: mdl-19234195

ABSTRACT

Thymic T cell development is dependent on a specialized epithelial microenvironment mainly composed of cortical and medullary thymic epithelial cells (TECs). The molecular programs governing the differentiation and maintenance of TECs remain largely unknown. Wnt signaling is central to the development and maintenance of several organ systems but a specific role of this pathway for thymus organogenesis has not yet been ascertained. In this report, we demonstrate that activation of the canonical Wnt signaling pathway by a stabilizing mutation of beta-catenin targeted exclusively to TECs changes the initial commitment of endodermal epithelia to a thymic cell fate. Consequently, the formation of a correctly composed and organized thymic microenvironment is prevented, thymic immigration of hematopoietic precursors is restricted, and intrathymic T cell differentiation is arrested at a very early developmental stage causing severe immunodeficiency. These results suggest that a precise regulation of canonical Wnt signaling in thymic epithelia is essential for normal thymus development and function.


Subject(s)
Cell Differentiation/immunology , Epithelial Cells/metabolism , Growth Inhibitors/genetics , Growth Inhibitors/metabolism , Thymus Gland/immunology , Thymus Gland/metabolism , beta Catenin/genetics , beta Catenin/metabolism , Animals , Cell Differentiation/genetics , Cell Line , Cells, Cultured , Endoderm/cytology , Endoderm/immunology , Endoderm/metabolism , Epithelial Cells/immunology , Epithelial Cells/pathology , Forkhead Transcription Factors/genetics , Gene Targeting , Growth Inhibitors/biosynthesis , Growth Inhibitors/physiology , Mice , Mice, Inbred C57BL , Mice, Transgenic , Signal Transduction/genetics , Signal Transduction/immunology , Thymus Gland/cytology , Thymus Gland/pathology , Wnt Proteins/genetics , Wnt Proteins/metabolism , beta Catenin/biosynthesis , beta Catenin/physiology
9.
Mech Dev ; 98(1-2): 111-4, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11044613

ABSTRACT

The expression pattern of the murine A33 antigen has been defined during development using wholemount immunohistochemistry. Two temporally and spatially distinct sites of expression were identified: the inner cell mass of the blastocyst and the endoderm cell layer of the intestinal tract where expression is initiated at E14.5 in the hindgut and subsequently extends throughout the length of the intestine. The onset of mA33 antigen expression in the gut occurs at the beginning of an extensive phase of cell movement involved in the conversion of the endoderm cell layer to a single cell layer of polarized epithelium. Expression of mA33 antigen is then maintained into adulthood, where it is a definitive marker of intestinal epithelium.


Subject(s)
Blastocyst/immunology , Intestines/embryology , Intestines/immunology , Membrane Glycoproteins/metabolism , Animals , Endoderm/immunology , Epithelium/embryology , Epithelium/immunology , Gene Expression Regulation, Developmental , Immunohistochemistry , Membrane Glycoproteins/genetics , Mice , Mice, Inbred ICR
10.
Mech Dev ; 49(1-2): 117-21, 1995 Jan.
Article in English | MEDLINE | ID: mdl-7538319

ABSTRACT

We describe here an antigen marker, designated VE-1, that is detected early in gastrulation (approximately E6.5 through approximately E7.25) in the anterior visceral endoderm overlying the embryonic ectoderm opposite the primitive streak. The antibody-positive domain extends from the embryonic-extraembryonic junction to the distal tip of the embryo, and laterally around approximately one-third of the circumference of the egg cylinder. Analysis of embryos at earlier stages indicates that VE-1 is first expressed shortly after implantation, at approximately E5.0, in the visceral endoderm on one side of the embryo and thus is the earliest molecular marker of A-P asymmetry in the post-implantation mouse embryo described to date. Although VE-1 was detected with a polyclonal antiserum raised against a 24 amino acid polypeptide sequence of FGF2, we provide evidence that the VE-1 antigen is not FGF2. The data reported here are the first to provide molecular evidence that A-P polarity in the mouse embryo is established by E5.0 and that the visceral endoderm has A-P polarity.


Subject(s)
Endoderm/immunology , Gastrula/immunology , Viscera/embryology , Animals , Antigens, Differentiation/analysis , Cell Polarity/immunology , Embryonic Development , Epitopes , Female , Mice , Pregnancy , Viscera/immunology
11.
Placenta ; 15(5): 525-39, 1994 Jul.
Article in English | MEDLINE | ID: mdl-7997452

ABSTRACT

An in vitro culture system has been devised creating apical and basal compartments separated by rabbit visceral yolk sac (VYS) with an intact epithelium. Selective transcytosis and binding of heterologous IgG applied to the apical yolk sac endoderm (YSE) was demonstrated in vitro using double label immunofluorescence. Thus, whilst both human and bovine IgG could be detected in endosomes in YSE, only human IgG could be detected in the basement membrane and vascular mesenchyme. This mirrors what is found in vivo. The Fc fragment of human Ig was transcytosed but not the Fab fragment, indicating that Fc receptors were expressed in the cultured YSE. When VYS was previously chilled to 4 degrees C to prevent endocytosis and treated with rabbit serum albumin to prevent non-specific binding, human IgG, but not bovine IgG, became specifically bound to YSE apical plasma membrane; comparison of binding at pH 6.0, 7.3 (the average pH of rabbit uterine fluid) and 8.0 revealed no obvious difference. Pre-exposure of VYS for up to 5 min in monensin, followed by culture in monensin and immunoglobulin-containing medium, did not prevent the selective transcystosis of human IgG, suggesting that an acidic compartment may not be needed for transcytosis. An acid pH dependent Fc gamma receptor equivalent to that on suckling rat gut jejunal enterocyte plasma membranes could not be isolated from rabbit YSE following exposure of solubilized membrane to affinity matrix bound IgG at pH 6.0 and elution at pH 8.0. These results contradict a recent suggestion that Fc receptors on all IgG transcytosing epithelia require an acid pH to effect IgG binding and selective transcytosis.


Subject(s)
Endoderm/metabolism , Immunoglobulin G/metabolism , Receptors, Fc/metabolism , Yolk Sac/metabolism , Animals , Cattle , Endocytosis , Endoderm/immunology , Endoderm/pathology , Female , Fluorescent Antibody Technique , Humans , Hydrogen-Ion Concentration , Immunoglobulin G/immunology , In Vitro Techniques , Middle Aged , Monensin/pharmacology , Protein Binding , Rabbits , Temperature , Yolk Sac/pathology
12.
J Histochem Cytochem ; 41(10): 1583-90, 1993 Oct.
Article in English | MEDLINE | ID: mdl-8245417

ABSTRACT

Our objective was to establish an immunohistological method for analysis of chimerism in mouse chimeras at embryonic stages with an anti-C3H strain-specific antigen (CSA) antibody. We developed an effective new method to retain CSA antigenicity with good morphology of embryonic tissues by using microwave irradiation (MWI) for pre-fixation, 95% ethanol/1% acetic acid as post-fixative solution, and polyester wax as embedding material. We used a biotinylated mouse monoclonal anti-CSA antibody, peroxidase-avidin, and silver amplification. These procedures were successful in demonstrating the chimerisms in various tissues of C3H<-->Balb/c chimeras at different embryonic stages and postnatal days. In chimeras at Days 7 and 7.5 post coitum (p.c.), both genotypes were clearly identified and well intermingling in every embryonic tissue (embryonic ectoderm, mesoderm, extra-embryonic ectoderm, ectoplacental cone, amnion, and chorion). Chimerisms at Day 14.5 p.c. were also clearly observed in mesencephalon, neural retina, spinal cord, lung, kidney, and liver. We concluded that the present immunohistological procedures for analysis of chimerism during embryonic periods will give us insightful information about dynamic histological changes such as cell proliferation, migration, selection, and death during organogenesis.


Subject(s)
Chimera , Fetus/cytology , Immunoenzyme Techniques , Amnion/cytology , Amnion/immunology , Animals , Antibodies, Monoclonal , Antigens/analysis , Antigens/immunology , Biotin , Chorion/cytology , Chorion/immunology , Ectoderm/cytology , Ectoderm/immunology , Endoderm/cytology , Endoderm/immunology , Fixatives , Genotype , Mesoderm/cytology , Mesoderm/immunology , Mice , Mice, Inbred BALB C , Mice, Inbred C3H , Microwaves , Silver Staining
13.
J Reprod Immunol ; 23(3): 247-64, 1993 Apr.
Article in English | MEDLINE | ID: mdl-8350300

ABSTRACT

The tissue distribution, molecular weight, and biochemical nature of an antigen detected by a mouse monoclonal antibody designated 283D3 and raised against rabbit visceral yolk sac endodermal cells, has been investigated. The antigen is located on the luminal side of apical tubules and large sub-apical vesicles in rabbit yolk sac endoderm and proximal kidney tubule epithelial cells. It is expressed in a similar polarised fashion in epithelial cells lining the epididymis. Western blotting showed the antigen to comprise proteins of molecular weight 330-380 kDa. The antigen has been affinity purified from yolk sac and kidney and is predominantly protein in nature with a small percentage of N-linked carbohydrate. In terms of tissue distribution and molecular weight it has close similarity to Heymann nephritis antigen but differs in not being confined to coated pits. Its function is not known, but the association with endocytic elements implies a possible role in non-specific protein absorption.


Subject(s)
Antigens, Surface/biosynthesis , Endoderm/immunology , Epididymis/immunology , Kidney Tubules, Proximal/immunology , Membrane Glycoproteins/biosynthesis , Rabbits/immunology , Yolk Sac/immunology , Animals , Antibodies, Monoclonal/immunology , Antigens, Surface/immunology , Antigens, Surface/isolation & purification , Blotting, Western , Cell Polarity , Endocytosis , Endoderm/metabolism , Epithelium/immunology , Female , Fluorescent Antibody Technique , Gene Expression , Immunohistochemistry , Kidney Tubules, Proximal/metabolism , Male , Membrane Glycoproteins/immunology , Membrane Glycoproteins/isolation & purification , Mice , Mice, Inbred BALB C/immunology , Molecular Weight , Organ Specificity , Yolk Sac/metabolism
14.
Kaibogaku Zasshi ; 66(2): 114-29, 1991 Apr.
Article in Japanese | MEDLINE | ID: mdl-1950430

ABSTRACT

Morphological and functional features of the yolk sac endodermal cells with special reference to the fetal macrophage differentiation were investigated morphologically under the light and electron microscopes and immunologically with the antigen phenotypic analysis and the phagocytic activity-test, using the syngeneic DA rat-embryos from 8 to 16 days of gestation. Based on the staining property with toluidin blue and the ultrastructural features, the endodermal layer from day 8 to 16-yolk sacs has been known to consist of two kinds of cell type; 10% "clear" cells with clear cytoplasm and 90% "dark" cells with dark cytoplasm. Numerous primary lysosomes, phagolysosomes, lipid droplets and coated vesicles distributed preferentially in the supranuclear portion of endodermal cells. A broad intercellular space was found between "clear" cells and "dark" cells, indicating the loose intercellular binding. It was often found that "clear" cells tend to migrate from the endodermal layer into the mesenchymal layer, where the poor development of basement membrane was seen between them. Cells phagocytosing red blood cells, that resemble morphologically "clear" cells, were also observed in the fetal liver. At ten hours after latex-injection into the yolk sac cavity of 14 days embryos, some cells which phagocytosed latex beads in their cytoplasm were found in the endodermal layer, and also in the liver tissue and loose connective tissue of fetus. These cells were stained positively with monoclonal antibody Mar3 which recognizes preferentially rat-mononuclear phagocyte system. In vitro-latex uptake of separated endodermal cells was also demonstrated by the culture-study of endodermal cell suspension. The present findings indicate that the yolk sac-endodermal layer derived from the proximal endoderm consists of at least two kinds of cell-population with a great similarity to tissue macrophages in morphological and functional senses, and support the concept that some cell-populations of endodermal layer may migrate into fetal tissue and are closely related to the differentiation of fetal macrophages and their precursors.


Subject(s)
Endoderm/cytology , Macrophages/cytology , Phagocytosis , Animals , Cell Differentiation , Endoderm/immunology , Endoderm/ultrastructure , Female , Macrophages/immunology , Microscopy, Electron , Pregnancy , Rats
15.
Differentiation ; 43(1): 1-9, 1990 Mar.
Article in English | MEDLINE | ID: mdl-1694800

ABSTRACT

XK endo B is a type I keratin that was originally identified by its preferential expression in the embryonic notochord of the amphibian Xenopus laevis. A peptide identical to a short region of its predicted amino acid sequence was used to generate antibodies against the XK endo B protein. This paper reports an immunocytochemical study of the spatial expression pattern of XK endo B during development. The protein was observed in the notochord and endoderm as predicted from previous RNA analysis. In addition, XK endo B was detected in the cement gland, in the pituitary, olfactory and pharyngeal pouch rudiments, and in a nonuniform distribution in the neural tube as well as the inner sensorial layer of the ectoderm. XK endo B expression is not limited to any germ layer or any particular cell type, but is nevertheless highly restricted in its distribution in the embryo. Its expression in several different embryonic tissues requiring inductive interactions for differentiation makes XK endo B a valuable tool with which to study the regulation of induced gene expression during embryogenesis.


Subject(s)
Ectoderm/metabolism , Embryo, Nonmammalian/metabolism , Endoderm/metabolism , Keratins/genetics , Notochord/metabolism , Xenopus laevis/embryology , Amino Acid Sequence , Animals , Antigens/immunology , Ectoderm/cytology , Ectoderm/immunology , Endoderm/cytology , Endoderm/immunology , Female , Fluorescent Antibody Technique , Gene Expression Regulation , Immune Sera/immunology , Keratins/immunology , Keratins/metabolism , Male , Notochord/cytology , Notochord/immunology , Pharynx/embryology , Pharynx/immunology , Pharynx/metabolism
16.
Histochemistry ; 94(5): 545-53, 1990.
Article in English | MEDLINE | ID: mdl-1704360

ABSTRACT

Mab 113F4, a monoclonal antibody recognizing an antigen in the outer synaptic layer of the chick neural retina, also recognizes an antigen appearing in all three germ layers of the gastrulating chick embryo. However, as neurulation proceeds, the antigen is down-regulated in three distinct patterns. First, the antigen is lost specifically from those trunk ectodermal cells destined to form the neural plate and, later, the neural tube. It remains absent from any neural derivative until day 13 when it appears in the outer synaptic layer of the neural retina, coincident with synaptogenesis in this region. Second, the entirety of the head ectoderm loses this antigen as the head lifts off the blastoderm. This down-regulation is followed later by a similar loss of antigen expression in the trunk ectoderm. Third, expression in the mesoderm becomes limited to the lateral plate and extraembryonic epithelia. Endodermal derivatives continue to express the antigen throughout development. Antigen 113F4 is localized within the cytoplasm and is organized in a fibrillar pattern. The intracellular localization of this antigen and its characteristic spatio-temporal tissue distribution are consistent with the antigen being a cytokeratin or cytokeratin-related antigen. The changes in tissue distribution suggest a possible role in tissue modelling in response to inductive interactions during development.


Subject(s)
Antibodies, Monoclonal/metabolism , Antigens/physiology , Chick Embryo/growth & development , Keratins/metabolism , Animals , Antibodies, Monoclonal/immunology , Antibody Specificity , Cell Differentiation , Chick Embryo/immunology , Chick Embryo/metabolism , Down-Regulation , Ectoderm/immunology , Ectoderm/metabolism , Endoderm/immunology , Endoderm/metabolism , Fluorescent Antibody Technique , Head/growth & development , Immunohistochemistry , Keratins/immunology , Mesoderm/immunology , Mesoderm/metabolism , Mice , Retina/growth & development , Retina/metabolism , Tissue Extracts/immunology
17.
Am J Obstet Gynecol ; 161(2): 462-7, 1989 Aug.
Article in English | MEDLINE | ID: mdl-2669495

ABSTRACT

The amnion is believed to be derived from either cytotrophoblastic cells or embryonic ectoderm. However, it produces and secretes CA 125, which is considered a differentiation antigen of fetal coelomic epithelium derived from the mesoderm of germ cells. To verify this, the immunohistochemical localization of CA 125 in human fetal tissues (between 7 and 23 weeks of gestation) derived from the ectoderm, endoderm, or mesoderm, and in the fetal membranes and placenta was studied. Among the mesoderm-derived tissues, only the fetal coelomic epithelium-related tissues were positive for anti-CA 125 from 15 weeks of gestation. The endoderm-derived tissues did not react with anti-CA 125. However, among the ectoderm-derived tissues, only the periderm reacted with anti-CA 125 from 7 weeks until it sloughed from the stratum intermedium by 23 weeks of gestation. Among the fetal membranes and placenta, only the amnion reacted with anti-CA 125 from 9 weeks to term. These findings indicate that the amnion and the periderm, both of which constitute the epithelia covering the amniotic cavity, in addition to the fetal coelomic epithelium-related tissues, produce CA 125.


Subject(s)
Amnion/immunology , Antigens, Tumor-Associated, Carbohydrate/analysis , Ectoderm/immunology , Endoderm/immunology , Epithelium/immunology , Extraembryonic Membranes/immunology , Female , Gestational Age , Humans , Immunoenzyme Techniques , Immunohistochemistry , Male , Mesoderm/immunology , Placenta/immunology , Pregnancy
18.
J Exp Zool ; 248(3): 315-20, 1988 Dec.
Article in English | MEDLINE | ID: mdl-3209980

ABSTRACT

Primary cultures of visceral yolk-sac (VYS) endodermal cells were used to assess the effects of teratogenic and nonteratogenic antibodies. When assessed by cytotoxicity assay, teratogenic antibodies appeared to be lethal to the cultured cells at high concentrations (1.25-5 mg of antibodies per ml of culture medium). At a nonlethal dosage, the teratogenic antibodies induced morphological changes, including retraction and rounding up of living cells. The cytotoxic effect as well as the effect on cell morphology appeared to be dose-dependent and specific to VYS endodermal cells. The mechanisms of cell killing were not the same as those attributed to complement-mediated cell lysis. The nonteratogenic antibodies did not have any cytotoxic effect nor did they cause any cell morphological alterations. The results of this investigation, when interpreted by correlating the dose-dependent effects of the teratogenic antibodies on cultured endodermal cells with the in vivo teratogenic effect, suggest that teratogenic antibodies when given at a teratogenic dose cause congenital abnormalities without killing the VYS endodermal cells.


Subject(s)
Antibodies/physiology , Endoderm/cytology , Teratogens , Yolk Sac/cytology , Animals , Antigens/immunology , Cell Survival , Cells, Cultured , Cytotoxicity Tests, Immunologic , Endoderm/immunology , Female , Rats , Rats, Inbred Strains , Yolk Sac/immunology
19.
Lab Invest ; 57(4): 350-8, 1987 Oct.
Article in English | MEDLINE | ID: mdl-2823004

ABSTRACT

We have developed two mouse monoclonal antibodies, M912-2A2 and M912-2G10, against cell surface antigens of a human infantile embryonal carcinoma cell line, MTE. The distribution of these antigens (designated as 2A2 and 2G10) was almost identical in human germ cell tumors in which they hallmarked yolk sac components and some tubular endodermal structures. Immunoelectron-microscopically, the antigens were located on the microvilli of MTE tumor cells. These antigens were not found on other common childhood tumors. In normal and fetal tissues they exhibited quite different distributions. In the kidney, 2A2 and 2G10 were present on the collecting tubules and proximal/distal tubules, respectively. Expression of both antigens was already observed in fetal kidneys of 10 weeks gestational age. In hematopoietic cells 2G10 was present only on granulocytes and on erythrocytes regardless of ABO blood group, whereas 2A2 was not present on any peripheral blood cells. Both antigens were equally expressed in testis and epididymis. Biochemically, reactivity of both antibodies was abolished with periodate treatment, suggesting their carbohydrate nature. Further biochemical characterization revealed that antibody to 2G10 reacts with the nonreducing terminal structure of type 2 carbohydrate chain, Ga1 beta 1-4G1cNAc, common to nLc4 (paragloboside), nLc6 (neolactohexaose), and Y4 neutral glycolipids of O-type erythrocytes. These data illustrate the complexity of carbohydrate antigens on yolk sac components of human germ cell tumors and provide a basis for the study of primitive endodermal and yolk sac differentiation in these tumors.


Subject(s)
Antibodies, Monoclonal , Antigens, Differentiation/analysis , Antigens, Neoplasm/analysis , Neoplasms, Germ Cell and Embryonal/immunology , Animals , Antibodies, Monoclonal/immunology , Antigens, Differentiation/immunology , Antigens, Neoplasm/immunology , Biomarkers, Tumor/analysis , Carbohydrates/immunology , Cell Membrane/immunology , Endoderm/immunology , Humans , Kidney/embryology , Kidney/immunology , Mice , Mice, Inbred BALB C , Microscopy, Electron , Neoplasm Transplantation , Neoplasms/immunology , Teratoma/immunology , Tissue Distribution , Tumor Cells, Cultured
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