Biomedical subjects

G Dohr

Publications and source records attributed to G Dohr.

At least 55 records · Page 3Linked to original sources

PubMed1997

Phenotypic comparison of natural killer cells from peripheral blood and from early pregnancy decidua.

In this descriptive flow cytometric study we analyzed the phenotype of human large granular lymphocytes from the decidua (DLGL) of first-trimester pregnancy. Expression of CD56 at high density on DLGL suggests a relationship to the small CD56bright+ subpopulation of peripheral blood natural killer (PBNK) cells. In comparison, these cell types differ in respect to the expression of a variety of adhesion molecules and receptors implicated in homing, migration and activation. In contrast to CD56bright+ PBNK cells, DLGL were still brighter for CD56 and show higher expression for CD29 and CD45RO. Less expression was found for CD15s, CD43, CD44, CD45RA, CD62L and HLA-DR. CD11a to c and CD18 were distributed in bimodal form on DLGL, part of the cells being negative. In summary, we found considerable differences between the cell surface marker profiles of DLGL and PBNK cells (subpopulations of the latter being separately analyzed).

Adolescent↗

PubMed1996

Expression of HLA class I molecules in human first trimester and term placenta trophoblast.

Expression of HLA class I molecules in trophoblast cells from various locations in normal human first trimester and term placenta was investigated by immunohistochemistry with a panel of monoclonal antibodies against the heavy chains or complete HLA class I molecules complexed with beta2-microglobulin. These reagents were also employed to distinguish between the products of different HLA class I loci. In addition to previously characterized reagents, a novel monoclonal antibody against HLA-A molecules (TU155) was used. Various choriocarcinoma and transfected cell lines served as controls for the specificities of the monoclonal antibodies. Cells in close contact with maternal cells, such as invading trophoblast cells and cells of the basal plate, expressed beta2-m micro globulin in association with HLA-G and HLA-C heavy chains. These class I heavy chains may also have been present as isolated molecules, although not in each of the cells. In contrast, cells of the chorion laeve exclusively expressed HLA-G, and not HLA-A, -B, or -C antigens. Our data support the often discussed immune protective function and the regulatory function of the HLA-G molecule, during invasion. In addition, by using monoclonal antibodies HCA2 (anti-HLA-A and -G), HC10 (anti-HLA-B and -C), TU149 (anti-HLA-B, -C, and some -A alleles), SFR8-B6 (anti-HLA-Bw6 and some -C), LA45 (some HLA-A and -B), TU48 (anti-HLA-Bw4 and some -A), and TU155 (anti-HLA-A), we show the presence of HLA-C molecules in all extravillous trophoblast cells of the cell columns and in the basal plate; the trophoblast cells of the chorion laeve lack this antigen. The function of this molecule is not clear, although a protective function against natural killer cell activity in the endometrium is postulated.

Blotting, Western↗

PubMed1996

The monoclonal antibody GZS-1 detects a maturation-associated antigen of human spermatozoa that is also present on the surface of human mononuclear blood cells.

A monoclonal antibody (GZS-1) has been generated by fusion of mouse myeloma cells with spleen cells from BALB/c mice immunised with human sperm cells. The antibody was determined to be an IgG1. The corresponding antigen is present on the whole surface of ejaculated human spermatozoa. It is not detectable on spermatozoa of other mammalian species (rabbit, cat, dog, sheep, boar, bull, horse). In human male genital organs, immunostaining with GZS-1 is observed on sperm cells in the epididymis and the ductus deferens together with the lining epithelium of those organs. No reactivity of sperm cells or germ cell precursors in the testis has been detected. Functional tests using the antibody show a strong inhibitory effect of human sperm in the hamster egg penetration assay. Furthermore, the GZS-1 antigen is detectable on the surface of human lymphocytes and monocytes by immunogold electron microscopy and FACS analysis. By Western blotting of human sperm and seminal plasma performed under reducing conditions immunostaining was detected at 21-25, 31, 51-54, and 62 kDa. The reaction with human lymphocytes shows one major band at 62 kDa and additional bands at 31 and 54 kDa. The results suggest that the monoclonal antibody GZS-1 may recognise an antigen which is secreted from the epithelial cells of the epididymis and binds to ejaculated spermatozoa as a sperm coating antigen. This component may be involved in the capacitation of the sperm and the acrosome reaction. Molecules that are expressed both on sperm and on immunocompetent cells may be relevant for the regulation of immunological processes or for the development of the related immunological tolerance of sperm in the female reproductive tract.

Antibodies, Monoclonal↗

PubMed1996

Differential phenotypic properties of human peripheral blood CD56dim+ and CD56bright+ natural killer cell subpopulations.

We investigated the presence of integrins and various other cell surface markers on the surface of freshly isolated CD56dim+ and CD56bright+ NK cells, the latter comprising a mean of 6.5% of total peripheral blood (PB) CD3-CD56+ NK cells. This small subpopulation stained more intensively for CD2, CD11c, CD15s, CD44, CD49e, CD55, CD62L, HLA-DR, and GZS-1, whereas there was no expression of CD49f in contrast to CD56dim+ cells. The myeloid antigen CDw65 was present on 11% of CD56bright+ cells, other myeloid markers were not found. 28% of CD56bright+ cells were positive for CD45R0. These results support the notion that CD56bright+ NK cells, based on their different marker profile, may possess different functional and biodistributional properties and--due to the signal-transducing capabilities of several adhesion molecules--differential patterns of stimulatability compared to the majority of PB-NK cells.

Adolescent↗

PubMed1996

Human first-trimester placenta intra-arterial trophoblast cells express the neural cell adhesion molecule.

The supposed influence of endometrial natural killer (NK) cells on the trophoblast invasion activities especially on intravasation of uteroplacental arteries in the non-pathogenic human first-trimester placenta was studied by means of immunohistochemistry. To identify extravillous trophoblast cells, smooth muscle cells, endothelia, endometrial glands, decidual stroma cells and endometrial NK cells, antibodies against cytokeratins, vimentin, smooth muscle cells, epithelium specific antigen and endothelial cells were employed. Furthermore, the immunohistochemical distribution patterns of CD56, CD57 and CD94 were studied and compared with the localization of invading trophoblast cells. Remodelling and dilatation of uteroplacental arteries starts before trophoblast cells can be found in the vicinity of the vessels. Nevertheless, subsequent trophoblast invasion of the arterial wall will lead to media destruction and intravasation only on focally restricted areas. This process is accompanied by the disappearance of endothelial cells and the immediate expression of the neural cell adhesion molecule (N-CAM, CD56) by intra-arterial trophoblast cells, which are eventually beginning to form intraluminal plugs. These findings led us to the conclusion that in the human pregnancy-induced physiological changes of the uteroplacental blood flow and the peripheral blood NK cell activity is not only, but also, due to the effect of CD56 expression by intra-arterial trophoblast cells.

Antibodies, Monoclonal↗

PubMed1995

Localisation of the high affinity facilitative glucose transporter protein GLUT 1 in the placenta of human, marmoset monkey (Callithrix jacchus) and rat at different developmental stages.

In the present study, the facilitative D-glucose transporter protein GLUT 1 was localised by immunohistochemistry in the placenta of human, marmoset (Callithrix jacchus) and rat at different developmental stages. A polyclonal antiserum against a 13-amino-acid peptide of the GLUT 1 carboxy terminus was used. It identified a protein of around 50 kDa molecular weight in immunoblotting of the placental tissues. GLUT 1 was located in the syncytiotrophoblast, in cytotrophoblast cells and in fetal endothelium. Similar staining patterns, except in human extravillous cytotrophoblast cells, were observed at all differentiation stages, despite differences in the internal placental architecture of the species. In the marmoset placenta, GLUT 1 was undetectable in endothelial cells of maternal vessels. In rat placentae, trophoblastic giant cells, epithelial cells of both visceral and parietal yolk sac, yolk sac vessels and the stratum spongiosum were stained. Reichert's membrane did not immunoreact. Preadsorption of the antiserum with a 13-amino-acid peptide resulted in the loss of immunoreactivity. The results suggest that GLUT 1 is a prominent isoform of glucose transporters in mammalian placentae. It is generally abundant in placental cell populations bordering on the maternal and fetal circulations and may therefore facilitate an effective glucose supply to the fetus and placenta.

Animals↗

PubMed1995

Immunohistochemical evidence of p53 protein in human placenta and choriocarcinoma cell lines.

We investigated the expression of the tumour-suppressor and cell cycle control protein p53 in human first trimester and term placenta, three choriocarcinoma cell lines (Jeg-3, JAR, BeWo) and human choriocarcinoma. Using monoclonal antibodies against p53 (DO-7, Ab-6, DO-1, PAb 1801), paraffin-embedded sections of first trimester and full-term placentae, human choriocarcinoma and Jeg-3, JAR and BeWo, as well as cytospins, were evaluated immunohistochemically. In addition, Western blots were carried out with the same antibodies on choriocarcinoma cell lines. In placentae, a small number of villous and extravillous cytotrophoblast cells, as well as very few syncytiotrophoblast cells, stained intensively. Also, p53 was visualized in some nuclei of the placental basal plate, whereas stroma and endothelium were negative for p53. Jeg-3, JAR and BeWo also showed a positive nuclear reaction with all applied antibodies. In paraffin-embedded sections of human choriocarcinoma, staining was confined to the nuclei of malignant cells. The results suggest that p53 is overexpressed not only in malignant tumour cells but in certain trophoblast cell populations of the human placenta as well.

Cell Line, Transformed↗

PubMed1995

Platelets contain interleukin-1 alpha and beta which are detectable on the cell surface after activation.

Activated platelets have been shown previously to exhibit membrane-bound IL-1 bioactivity, which leads to the question of localization of the cytokine in platelets. Using immunocytological and flow cytometric techniques, we found IL-1 alpha and IL-1 beta in the cytoplasma of both resting and thrombin-activated platelets. Immunogold-silver staining of the cell surface of activated platelets as well as preembedding antibody treatment of platelets revealed the presence of IL-1 (alpha and beta) in low density on the surface of intact cells in contrast to distinct enrichment in the cytoplasma of damaged platelets. Fibrin fibres present between cells indicated adsorbance of IL-1. There was also weak binding of anti-IL-1 alpha to the surface of thrombin-activated platelets as shown by flow cytometry. Following activation there appears to be some transfer of IL-1 onto the cell surface of activated cells, the bulk of the cytokine, however, is probably not released prior to platelet disintegration. In summary, we present evidence for the presence of both IL-1 alpha and IL-1 beta in resting and activated platelets without being able to demonstrate localization of the cytokines to specific subcellular structures.

Antigens, Surface↗

PubMed1995

Generation, characterization, and histochemical application of monoclonal antibodies selectively recognizing oxidatively modified apoB-containing serum lipoproteins.

To investigate either the role oxidized LDL plays in atherosclerosis or structural changes on the surface of oxidized LDL, monoclonal antibodies (mAbs) are an important tool. After immunizing mice with Cu(2+)-oxidized LDL (oxLDL) and fusion of splenocytes, hybridoma supernatants were screened and cloned. Two mAbs, OB/04 and OB/09 (IgG and IgM), were further characterized. In solid-phase fluorescence immunoassays and Western blot analysis both mAbs reacted with oxLDL, LDL oxidized by a free radical-generating azo compound, or oxVLDL but not with native LDL, acetylated LDL, oxHDL3, azo-oxidized HDL3, or HDL3 modified with malondialdehyde (MDA). In competitive immunoassays with LDL modified by oxidized fatty acid-derived aldehydes, mAb OB/09 strongly reacted with MDA-LDL or MDA-VLDL and LDL modified with 4-hydroxyhexenal followed by 4-hydroxynonenal but not with 4-hydroxyoctenal or hepta-2,4-dienal. mAb OB/04 had a weak affinity for LDL after modification with these aldehydes except for MDA-LDL. LDL modified with arachidonic acid oxidation products (AAOPs) was also recognized by this mAb. However, albumin modified either by the aldehydes applied or by AAOPs did not react with either mAb. Thus, the data indicate that each of the mAbs recognizes a different epitope that is expressed only on apoB-containing lipoproteins upon oxidative modification. An immunostaining with mAb OB/04 was obtained in areas rich in macrophages and in connective tissue of a human atherosclerotic lesion.

Amidines↗

PubMed1995

[Demonstration of biophysical effects of pulsed laser irradiation with histological tissue sections].

For therapeutic application of laser light it is necessary to minimize defects in the nonirradiated tissue. These defects depend on the primary mechanism of interaction between tissue and laser light. Three experiments were performed to distinguish between mechanical and thermal effects of nano- and microsecond laser pulses in skeletal muscle of the rat. The light, transmission and scanning electron microscopes were In the ns-experiments the mechanical action of a single ns pulse (8 ns) produced a crater. Only zones I and IV developed. With 50 to 100 pulses all zones can be identified. These results show that a single ns pulse suffices to form a tissue crater by mechanical action. A higher number of ns pulses leads to heat accumulation and produces thermal lesions similar to those seen after application of microseconds-pulses.

Animals↗

PubMed1994

Insulin receptors in syncytiotrophoblast and fetal endothelium of human placenta. Immunohistochemical evidence for developmental changes in distribution pattern.

The localisation of insulin receptors (IR) was investigated on cryosections of human non-pathologic first trimester and full term placentae by indirect immunohistochemistry with three different monoclonal antibodies (MABS). In placentae from 6 to 10 weeks postmenstruation (p-m.), only syncytiotrophoblast was stained, predominantly that of mesenchymal villi and syncytial sprouts, which are areas of high proliferative activity. In placentae from 11 to 14 weeks p-m., endothelial cells commenced to react with the IR MABS and the syncytiotrophoblast was less intensely labelled than at weeks 6 to 10 p-m. In term placentae, the microvillous membrane of the syncytiotrophoblast showed only patches of weak immunoreactivity. In contrast, the endothelial cells in the placenta but not in the umbilical cord were strongly stained. The amniotic epithelium in the chorionic plate and fibroblasts in the stroma were conspicuously labelled. The data indicate: (1) the receptor density on villous syncytiotrophoblast decreases and that of fetal endothelium increases' throughout gestation; (2) syncytiotrophoblast of human term placentae expresses a low level per unit area of surface IR; and (3) the majority of IR in human term placentae is located in fetal endothelium. Apart from yet unknown functional effects of maternal and fetal insulin at the placental barrier, the results suggest a growth promoting effect on the trophoblast of maternal insulin in first trimester as well as developmental effects of fetal insulin on the feto-placental vessels at term.

Antibodies, Monoclonal↗

PubMed1994

Heterogeneous histochemical reaction pattern of the lectin Bandeiraea (Griffonia) simplicifolia with blood vessels of human full-term placenta.

Bandeiraea simplicifolia lectin (BS-I) stains vascular endothelium in various species. In humans, less than 10% of the specimens studied exhibit a reaction with BS-I. In the present histochemical study, the reactivity of BS-I with placental blood vessels and its correlation with the blood group from mother and newborn child was investigated. Acetone-fixed cryosections of representative tissue segments of human full-term placenta and umbilical cord were stained with BS-I. The staining pattern of tissues from patients with different blood groups was identical, although the reaction of BS-I in the placenta was heterogeneous. BS-I did not react with the umbilical cord. Vascular smooth muscle cells at the insertion site of the umbilical cord into the chorionic plate, and endothelium deeper in the chorionic plate, became progressively stained. The endothelial cells and tunica muscularis of smaller arteries and veins in stem villi lost their reactivity in parallel with decreasing vessel size. Arterioles and venules reacted heterogeneously. Capillaries, trophoblastic basement membranes, especially epithelial plates, and sometimes the syncytiotrophoblast were labelled in several terminal villi. The data indicate that 1) the placenta binds BS-I to fetal endothelium independent of the blood group, 2) cell-surface antigens on placental endothelial cells are expressed heterogeneously and 3) cell-surface glycans are constituted in an organ-specific manner on human endothelial cells.

Endothelium, Vascular↗

PubMed1994

Differential lectin binding to the fibrinoid of human full-term placenta: correlation with a fibrin antibody and the PAF-Halmi method.

Placental fibrinoid is thought to contain various glycoproteins originating from cell secretion and tissue degeneration, occasionally merged with fibrin. Information on the characteristics and derivation of the various fibrinoid components, however, is still fragmentary. Therefore, the present histochemical study on acetone-fixed placental tissue sections compared the staining pattern of FITC-conjugated lectins from Ulex europaeus (UEA-I), Bandeiraea simplicifolia (BS-I) and Lycopersicon esculentum (LEA) with the reactivity of a fibrin antibody and a modified paraldehyde-fuchsin stain. Using different color reactions, the latter histological method identified two types of fibrinoid, which correlated well with fibrin-type and matrix-type fibrinoid. Immunohistochemically, fibrin was detected at the intervillous border of the basal plate, in some inner parts and in perivillous fibrinoid. UEA-I bound to endothelial cells and partially to fibrin-type fibrinoid of villi and the basal plate, thus indicating a reaction with immured and disintegrated remnants of endothelial and blood cells. LEA stained fibrin-negative (i.e. matrix-type) fibrinoid homogeneously within the basal plate; the small reactive areas within the perivillous fibrinoid may belong to degenerating trophoblastic residues, because in villi LEA specifically reacted with the syncytiotrophoblast. BS-I heterogeneously labeled matrix-type fibrinoid deposits in the basal plate which surrounded decidual cells and subpopulations of extravillous trophoblast. In cell islands, BS-I also stained fibrinoid surrounding trophoblast cells heterogeneously. The data (1) confirm the existence of two types of placental fibrinoid, fibrin-type and matrix-type fibrinoid, (2) suggest that both types of fibrinoid contain different glycoconjugates, and (3) demonstrate the practical usefulness of the modified paraldehyde-fuchsin method for the identification of the two types of fibrinoid.

Endothelium, Vascular↗

PubMed1994

Non-Michaelis-Menten kinetics of zero-trans glucose uptake by trophoblast cells from human term placentae and by choriocarcinoma (JEG-3/JAR) cells.

Maternal glucose is a major substrate for placental and fetal metabolism. The kinetics of its uptake into placental trophoblast cells has not been characterised yet and was therefore investigated in the present study. In addition to trophoblast cells isolated from human term placentae, JEG-3 and JAR choriocarcinoma cells were used. Measurements were carried out in 5 s intervals until 30 s with the non-metabolisable glucose analogue 3-O-[14C]methyl-D-glucose using confluent cells adhering to glass coverslips. L-[1-14C]glucose was used to correct for extracellular trapped tracer and diffusion. The uptake was rapid and saturable. It reached equilibrium after 30 s at 20 degrees C and could be inhibited by 0.4 mmol/l cytochalasin B up to 98%. The choriocarcinoma cells took up twice as much glucose as trophoblast cells. Fitting the experimental data to the Michaelis-Menten equation by non-linear regression failed to adequately describe the data, even when a contribution of diffusion to total uptake was considered. Introducing the Hill coefficient n into the Michaelis-Menten equation significantly improved the quality of the fits as was assessed by three statistical criteria. Using this equation modified for allosteric kinetics (v = k[To] [S]n)/(Km + [S]n)), parameters were calculated as Km = 12 mmol/l, Vmax = 17 fmol/l s-1 per cell, n = 1.1 for trophoblast cells; Km = 13 mmol/l, Vmax = 27 fmol/l s-1 per cell, n = 1.2 for JEG-3 cells and Km = 29 mmol/l, Vmax = fmol/l s-1 per cell, n = 1.4 for JAR cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Adhesion↗

PubMed1993

Expression of decay-accelerating factor (CD55) of the complement system on human spermatozoa.

Human spermatozoa were analyzed for their expression of decay-accelerating factor (DAF, CD55), a glycolipid-anchored regulatory protein of the C casade. Morphologic data showed that DAF was localized at the acrosomal region of the sperm head. Analysis with anti-DAF antibody-immunoprecipitated proteins of acrosome-reached spermatozoa revealed a 44- to 54-kDa protein. Carbohydrate analysis of sperm DAF indicated that it contains nonsialated N- and O-linked sugars. The absence of mature oligosaccharides on this protein appears to account for the difference in molecular mass between sperm DAF and the 70-kDa DAF expressed on other human tissues. Sperm DAF reinserted into sheep E and inhibited C-mediated lysis. This effect was reversed by mAb, which block DAF function. These results indicate that sperm DAF also possesses a glycolipid anchor. The expression of DAF on acrosome-reacted spermatozoa suggests that it may act concomitantly with other C regulators such as membrane cofactor protein to modulate the activation of C in the immunocompetent female genital tract and protect acrosome-reacted spermatozoa from C-mediated attack.

Acrosome↗

PubMed1993

Immunohistochemical evidence for the heterogeneity of maternal and fetal vascular endothelial cells in human full-term placenta.

The heterogeneity of endothelial cell surface antigen expression was studied in 5 human full-term placentae by means of indirect immunohistochemistry using 9 monoclonal antibodies and by staining with fluorescent-conjugated Ulex europaeus lectin, both of which are widely used endothelial cell markers. (1) A highly specific, homogeneous staining of fetal and maternal placental vessels of all sizes and anatomical regions was observed by the monoclonal antibodies PAL-E, QBEND10 and 1F10. These antibodies were even more specific than Ulex europaeus lectin, factor VIII antibody and von Willebrand factor antibody, which cross-reacted with some non-endothelial cells and structures. The reactivity of PAL-E, QBEND10 and 1F10 with residual surface cells of the basal plate strongly suggests an endothelial origin of these cells. (2) In contrast to other organs, PAL-E, QBEND10 and HM15/3 strongly stained endothelial cells of the macrovascular system in the human placenta. This might indicate an organ-associated heterogeneity of fetal endothelial cells. (3) Monoclonal antibodies against receptors for transferrin and IgG (Fc gamma RII) labeled the endothelial cells of fetal placental vessels with increasing intensity distal to the insertion of the umbilical cord. The vessels of the umbilical cord itself were unreactive. This might suggest a heterogeneity of macro- and microvascular endothelial cells.

Antibodies, Monoclonal↗

PubMed1993

Expression of various MHC class II molecules and of intracellular adhesion molecule-1 (ICAM-1) on focal clusters of dendritic cells in iodine deficiency goitres.

Thyroid sections from 18 consecutive euthyroid patients undergoing surgery for iodine deficiency goitre were investigated by means of immunohistochemistry and immunofluorescence, evaluating the expression of MHC class II antigens (HLA-DR, -DP, -DQ, and RFD1) and intercellular adhesion molecule-1 on the formerly described clusters of dendritic cells, as well as on thyrocytes. Eleven of 18 iodine deficiency goitres contained clusters of dendritic cells. These clusters appeared to express only HLA-DR in two cases; in nine of 12 cases they showed a differential expression of class II molecules in the following frequency: HLA-DR > DQ and/or -DP > RFD1. These dendritic cells also were ICAM-1+. In four of 18 iodine deficiency goitres, thyroid epithelial cells showed MHC class II expression in several combinations, but were ICAM-1-. In normal thyroids and in nodular goitres from inhabitants of the endemic area not having an actual iodine deficiency, only sparse clusters of dendritic cells were found; these cells were only HLA-DR+. Follicle lining cells were negative for the MHC class II molecules. In normal thyroids from an area with sufficient iodine supply, no clusters of dendritic cells were seen. The few dendritic cells observed were lying isolated in the interstitium and only positive for HLA-DR and ICAM-1; epithelial cells were negative for the studied markers. These data show clusters of dendritic cells in thyroids of inhabitants of an endemic area. When goitre is accompanied by iodine deficiency at the moment of operation, there appears to be activation of these dendritic cells and of thyroid epithelial cells.

Adult↗

PubMed1990

Localization and characterization of the acrosomal antigen recognized by GB24 on human spermatozoa.

GB24, a mouse monoclonal antibody, recognizes a trophoblast-leukocyte cross-reactive antigen (TLX), which is likely identical to the membrane cofactor protein (MCP), a complement regulatory protein. GB24 reacts also with a human acrosomal sperm antigen (Fénichel et al.: J Reprod Fertil 87:699-706, 1989). By immunofluorescence or immunoperoxidase, testicular, epididymal, and ejaculated spermatozoa were found to be positive after fixation by acetone. Motile, suspended spermatozoa became positive only through conditions known to induce acrosome reaction (A23187, follicular fluid, contact with oocytes). Ultrastructural studies with immunogold staining localized this protein on the inner acrosome membrane and in the acrosomal content. By SDS-polyacrylamide gel electrophoresis, GB24 immunoprecipitated a unique protein of 48 kDa from capacitated and A23187-induced spermatozoa under reducing conditions. No cross-reactivity was found with mouse, boar, or ram spermatozoa. Localization of this human sperm antigen recognized by GB24 and its similarity with the TLX-MCP family antigens would suggest a possible role of this molecule during fertilization in sperm-egg binding or immune protection.

Acrosome↗