scholarly journals A self-marker-like protein governs hemocyte allorecognition in Halocynthia roretzi

2019 ◽  
Vol 5 (1) ◽  
Author(s):  
Masaki Ema ◽  
Taizo Okada ◽  
Miki Takahashi ◽  
Masato Uchiyama ◽  
Hideo Kubo ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Single Gene ◽  
Phenol Oxidase ◽  
2D Electrophoresis ◽  
Fusion Reactions ◽  
Halocynthia Roretzi ◽  
Primitive Form ◽  
Marker Proteins ◽  
Isoelectric Points ◽  
Reaction Characteristic

Abstract Background Self-incompatibility, fusion/non-fusion reactions, and contact reactions (CRs) have all been identified as allorecognition phenomena in ascidians. CR is a reaction characteristic of the hemocytes of Halocynthia roretzi, whereby they release phenol oxidase (PO) upon contact with non-self hemocytes. Thus, these cells may represent a primitive form of the vertebrate immune system. In the present study, we focused on the CR of H. roretzi hemocytes and sought to identify self-marker proteins that distinguish between self and non-self cells. Results We initially generated a CR-inducing monoclonal antibody against the complete hemocyte membrane-protein complement (mAb11B16B10). This antibody was identified based on the differential induction of PO activity in individual organisms. The level of PO activity induced by this antibody in individual ascidians was consistent with the observed CR-induced PO activity. mAb11B16B10 recognized a series of 12 spots corresponding to a 100-kDa protein, with differing isoelectric points (pIs). A comparison of the 2D electrophoresis gels of samples from CR-reactive/non-reactive individuals revealed that some spots in this series in hemocytes were common to the CR-non-inducible individuals, but not to CR-inducible individuals. We cloned the corresponding gene and named it Halocynthia roretzi self-marker-like protein-1 (HrSMLP1). This gene is similar to the glycoprotein DD3–3 found in Dictyostelium, and is conserved in invertebrates. Conclusion We generated a CR-inducing monoclonal antibody (mAb11B16B10) that recognized a series of novel membrane proteins (HrSMLP1) in the hemocytes of H. roretzi. The combination of expressed spots of HrSMLP1 distinguishes non-self cells from self cells with respect to CR inducibility. Given that the HrSMLP1 gene is a single gene, it may represent a novel type of self-marker protein with a role in CR.

scholarly journals Characterization of Inhibin from Culture and Non Culture of Granulose Cells for Monoclonal Antibody of Inhibin Production

2010 ◽  
Vol 4 (1) ◽  
Author(s):  
Amiruddin Amiruddin ◽  
Tongku Nizwan Siregar ◽  
Amalia Sutriana ◽  
Dwinna Aliza ◽  
T. Armansyah
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Western Blot ◽  
Total Amino Acid ◽  
Multiple Ovulation ◽  
Sds Page ◽  
Isoelectric Points ◽  
Granulose Cells

This study has long-term objectives to obtain immunogenic prototype that can be used to induce multiple ovulation in goats. Working steps of this study were begun with the collection of ovarium from goats, collection of granulose cells, culture of granulose and characterization of molecular weight and isoelectric point (pI) of inhibin protein of granulose cells obtained from culture and non-culture of granulose cells, and followed by preparation of monoclonal antibody toward inhibin. The results showed that inhibin isolated either from culture or non-culture of granulose cells produced a 32 kDa band. Molecular weight of inhibin was measured by Western Blot. The 32 kDa band of SDS PAGE product appeared on Western Blot result was inhibin molecules produced by granulose cells collected fom culture and non-culture of granulose cells that can be identified by Mab-inhibin. Product of IEF gel electrophoresis suggested that inhibin molecule collected from culture of granulose cells has no charge at isoelectric points ranging from 5-6, depends on its total amino acid composition.

Variation in cytosolic protein expression between human colon tumors that differ with regard to differentiation class.

1988 ◽  
Vol 34 (1) ◽  
pp. 71-75 ◽  
Author(s):  
T J Nalty ◽  
C W Taylor ◽  
L C Yeoman
Keyword(s):  
Human Colon ◽  
Human Colon Cancer ◽  
Two Dimensional Gel Electrophoresis ◽  
Colon Tumors ◽  
Marker Proteins ◽  
Isoelectric Points ◽  
Molecular Masses ◽  
Poorly Differentiated ◽  
Differentiation Marker ◽  
Individual Differentiation

Abstract Using a combination of two-dimensional gel electrophoresis, silver staining, and a 16-quadrant grid system, we established a set of composite patterns for the colon tumor cytosol proteins of well, moderately, and poorly differentiated tumors. These composite patterns were found to be characteristic of the three individual differentiation classes for colon tumors. The apparent relative molecular masses (Mr) of the resolved proteins ranged from 14,000 to 105,000 and their isoelectric points from pH 5.2 to 8.4. Although the vast majority of the proteins identified in the composite patterns were common, a comparison based upon these patterns revealed two qualitative and seven quantitative protein differences. The qualitative differences were identified by apparent Mr X 10(-3)/pI coordinates of 73/7.2 and 66/6.2. Quantitative differences were identified by Mr X 10(-3)/pI coordinates of 71/6.0, 59/6.7, 57/6.7, 56/5.4, 52/6.1, 30/5.8, and 18/6.2. These cytosolic differentiation marker proteins may facilitate the diagnosis, staging, and monitoring of human colon cancer.

scholarly journals Novel mesenchymal and haematopoietic cell isoforms of the SHP-2 docking receptor, PZR: identification, molecular cloning and effects on cell migration

2003 ◽  
Vol 370 (2) ◽  
pp. 537-549 ◽  
Author(s):  
Andrew C.W. ZANNETTINO ◽  
Maria ROUBELAKIS ◽  
Katie J. WELLDON ◽  
Denise E. JACKSON ◽  
Paul J. SIMMONS ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Single Gene ◽  
Transcription Unit ◽  
Erythroid Precursor ◽  
Haematopoietic Cells ◽  
Colony Forming Units ◽  
Cytoplasmic Tails ◽  
Src Homology ◽  
First Time

SHP-2 (Src homology phosphatase type-2) is essential for haematopoietic skeletal and vascular development. Thus the identification of its binding partners is critically important. In the present study, we describe a unique monoclonal antibody, WM78, which interacts with PZR, a SHP-2 binding partner. Furthermore, we identify two novel isoforms of PZR, PZRa and PZRb, derived by differential splicing from a single gene transcription unit on human chromosome 1q24. All are type 1 transmembrane glycoproteins with identical extracellular and transmembrane domains, but differ in their cytoplasmic tails. The PZR intracellular domain contains two SHP-2 binding immunoreceptor tyrosine-based inhibitory motifs (VIY246AQL and VVY263ADI) which are not present in PZRa and PZRb. Using the WM78 monoclonal antibody, which recognizes the common extracellular domain of the PZR isoforms, we demonstrate that the PZR molecules are expressed on mesenchymal and haematopoietic cells, being present on the majority of CD34+CD38+ and early clonogenic progenitors, and at lower levels on CD34+CD38- cells and the hierarchically more primitive pre-colony forming units. Interestingly, we show by reverse transcriptase-PCR that the PZR isoforms are differentially expressed in haematopoietic, endothelial and mesenchymal cells. Both PZR and PZRb are present in CD133+ precursors and endothelial cells, PZRb predominates in mesenchymal and committed myelomonocytic progenitor cells, and all three isoforms occur in erythroid precursor cell lines. Importantly, using SHP-2 mutant (Δ46—110) and SHP-2 rescue of embryonic fibroblasts stably expressing the PZR isoforms, we demonstrate for the first time that PZR, but not PZRa or PZRb, facilitates fibronectin- dependent migration of cells expressing a competent SHP-2 molecule. These observations will be instrumental in determining the mechanisms whereby PZR isoforms regulate cell motility.

scholarly journals Antigens of Adults and Third-Stage Larvae of the Meningeal Worm, Parelaphostrongylus Tenuis (Nematoda, Metastrongyloidea)

1994 ◽  
Vol 6 (2) ◽  
pp. 222-229 ◽  
Author(s):  
N. F. Neumann ◽  
W. S. Pon ◽  
A. Nowicki ◽  
W. M. Samuel ◽  
M. Belosevic
Keyword(s):  
Trichinella Spiralis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
2D Electrophoresis ◽  
Dictyocaulus Viviparus ◽  
Sds Page ◽  
Third Stage ◽  
Parelaphostrongylus Tenuis ◽  
Isoelectric Points ◽  
Initial Results

We studied the antigens of adults and third-stage larvae of the meningeal worm, Parelaphostrongylus tenuis, in an attempt to identify potential serodiagnostic molecules for this important infection of wild ungulates. Soluble extracts of P. tenuis adult worms and third-stage larvae were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional (2D) electrophoresis and analyzed by immunoblotting using purified rabbit anti- P. tenuis immunoglobulin G (IgG). The IgG antibodies were obtained from animals immunized with P. tenuis adult worm or third-stage larva soluble extract and serum from elk infected with P. tenuis. Out of more than 75 antigens (as shown by 2D electrophoresis and immunoblotting), 7 antigens from adults (four 170-120-kD molecules with isoelectric points between 6.0 and 6.6, two 55kD molecules with isoelectric points of 5.6 and 5.8, and one 13-kD molecule) and 2 antigens from third-stage larvae (one 25-30-kD molecule with an isoelectric point of 6.3 and one 13-kD molecule) distinguished P. tenuis from two other nematodes, Dictyocaulus viviparus and Trichinella spiralis. Initial results using serum from experimentally infected elk indicate that this serum recognized a similar profile of P. tenuis antigens when compared with the serum from immunized rabbits. This research has set the foundation for the development of a test for P. tenuis infections in wild and recently domesticated elk and other ungulates.

Characterization of major surface and excretory-secretory immunogens ofTrypanosoma cruzitrypomastigotes and identification of potential protective antigen

Parasitology ◽  
1990 ◽  
Vol 100 (1) ◽  
pp. 115-124 ◽  
Author(s):  
M. A. Ouaissi ◽  
A. Taibi ◽  
J. Cornette ◽  
P. Velge ◽  
B. Marty ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Dimensional Analysis ◽  
Polyclonal Antibody ◽  
Protective Antigen ◽  
Surface Antigens ◽  
Antigenic Structure ◽  
Enzyme Linked Immunosorbent Assay ◽  
Two Dimensional ◽  
Wide Range ◽  
Isoelectric Points

SummaryThe surface antigens ofTrypanosoma cruzitrypomastigotes were identified by immunoprecipitation and were compared with metabolically labelled excretory—secretory products (ES) released by the parasitesin vitro. A series of major immunogenic components in the ES antigens were revealed (160 kDa, 130 kDa and 80–110 kDa). The trypomastigote surface also bears the 130 kDa band and the 80–110 kDa complex. Competition experiments demonstrated the common antigenic structure of the ES and the surface antigens. Two-dimensional analysis of ES antigens immunoprecipitated by human Chagasic serum revealed several spots in the 80–110 kDa region with a wide range of isoelectric points (PI between 5·4 and 6·7). This reflects a charge heterogeneity of these polypeptides. The trypomastigote 85 kDa polypeptide was also identified in the ES antigens by using a monoclonal antibody against this antigen. Two-dimensional analysis of the 85 kDa proteins shed from the surface of trypomastigotes and immunoprecipitated by the monoclonal antibody 155D3 showed 2 major spots: a major part of the 85 kDa polypeptide was found at pH 6·5–6·6, whereas a substantial amount of the antigen was found at pH 5·7. An additional component with molecular weight of approximately 58 kDa and isoelectric points of 6·5 and 6·6, was also visualized. Detection of the 85 kDa polypeptide circulating in serum from patients with acute and chronic Chagas' disease was achieved using an enzyme-linked immunosorbent assay. In addition, the data obtained showed that a polyclonal antibody to the 85 kDa polypeptide could be used to passively induce a partial protection of Fischer rats against acute lethal infection. Thus, the antigens recognized by polyclonal antibody appear to play a role in the development of protective immunity againstT. cruzi.

Differential expression and secretion of α1-acid glycoprotein in bovine milk

2007 ◽  
Vol 74 (3) ◽  
pp. 374-380 ◽  
Author(s):  
Fabrizio Ceciliani ◽  
Vanessa Pocacqua ◽  
Cristina Lecchi ◽  
Riccardo Fortin ◽  
Raffaella Rebucci ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Differential Expression ◽  
Main Part ◽  
Lectin Binding ◽  
Bovine Milk ◽  
Mammary Tissue ◽  
2D Electrophoresis ◽  
Acid Glycoprotein ◽  
Isoelectric Points ◽  
Glycan Pattern

α1-Acid glycoprotein (AGP) is a lipocalin that is produced mainly by the liver and secreted into plasma in response to infections and injuries. In this study, we evaluated AGP isoforms that can be detected in bovine milk. We found that milk-AGP content is made up of at least two isoform groups, a low MW group (44 kDa) that is produced in the mammary gland (MG-AGP), and a higher MW group (55–70 kDa), that is produced by somatic cells (SC-AGP). Identical SC-AGP isoforms can be found both in milk and blood PMN cells. Analysis of the mammary tissue cDNA showed that the sequence of the MG-AGP isoform is identical to that of plasma AGP. Each group contains several proteins with different MWs and different isoelectric points, as shown by 2D-electrophoresis. The glycosylation patterns of these isoforms were analysed by means of specific lectin binding, to evaluate the degree of sialylation, fucosylation and branching. The MG-AGP glycan pattern was identical to plasma AGP produced by the liver. Several differences were detected, however, between plasma and SC-AGP isoforms, the most evident being the strong degree of fucosylation and the elevated number of di-antennary glycans in SC-AGP. Immunohistochemistry showed that AGP is found in all tissues that make up the mammary gland, but that it is most likely produced for the main part by the alveoli.

scholarly journals Biosynthesis of Pb44, the protective antigen of sporozoites of Plasmodium berghei.

1981 ◽  
Vol 154 (4) ◽  
pp. 1225-1236 ◽  
Author(s):  
N Yoshida ◽  
P Potocnjak ◽  
V Nussenzweig ◽  
R S Nussenzweig
Keyword(s):  
Monoclonal Antibody ◽  
Protein Synthesis ◽  
Salivary Gland ◽  
Cell Membrane ◽  
Plasmodium Berghei ◽  
Protective Antigen ◽  
Surface Membrane ◽  
Molecular Weights ◽  
Isoelectric Points ◽  
The Relationship

In a previous paper (2) we identified a protective antigen (Pb44) of the surface membrane of sporozoites of Plasmodium berghei by means of a monoclonal antibody. Immunoprecipitation of extracts of mature salivary gland sporozoites, metabolically labeled with L[35S]methionine using the same monoclonal antibody, revealed three specific polypeptides: *Pb44, *Pb52, and *Pb54. Metabolically labeled *Pb44 is probably identical to the protective antigen previously identified by surface labeling. Both proteins have the same molecular weights and isoelectric points under denaturing conditions, and they share an epitope. Moreover, *Pb44 also seems to be located on the cell membrane. The results of pulse-chase experiments strongly suggest that *Pb52 is the precursor of *Pb44. The relationship between *Pb54 and the protective antigen is unknown. The three polypeptides seem to be strictly associated with only one of the developmental stage of the parasite. They were not detected in blood forms and were found in minute amounts in sporozoites from the midgut of mosquitoes. In contrast, in mature salivary gland sporozoites they constitute main products of protein synthesis.

Exsolution and inversion in pigeonite from the Whin Sill, Northern England

1978 ◽  
Vol 36 (1) ◽  
pp. 474-475
Author(s):  
P.P.K. Smith
Keyword(s):  
High Temperature ◽  
Low Temperature ◽  
Homogeneous Nucleation ◽  
Silicate Mineral ◽  
Antiphase Boundaries ◽  
Two Phase ◽  
Primitive Form ◽  
The Core ◽  
Nucleation Mechanisms ◽  
And Inversion

Grains of pigeonite, a calcium-poor silicate mineral of the pyroxene group, from the Whin Sill dolerite have been ion-thinned and examined by TEM. The pigeonite is strongly zoned chemically from the composition Wo8En64FS28 in the core to Wo13En34FS53 at the rim. Two phase transformations have occurred during the cooling of this pigeonite:- exsolution of augite, a more calcic pyroxene, and inversion of the pigeonite from the high- temperature C face-centred form to the low-temperature primitive form, with the formation of antiphase boundaries (APB's). Different sequences of these exsolution and inversion reactions, together with different nucleation mechanisms of the augite, have created three distinct microstructures depending on the position in the grain.In the core of the grains small platelets of augite about 0.02μm thick have farmed parallel to the (001) plane (Fig. 1). These are thought to have exsolved by homogeneous nucleation. Subsequently the inversion of the pigeonite has led to the creation of APB's.

A web-like network of type vi collagen in human skin is revealed by immuno-electron microscopy

1988 ◽  
Vol 46 ◽  
pp. 382-383
Author(s):  
Douglas R. Keene ◽  
Robert W. Glanville ◽  
Eva Engvall
Keyword(s):  
Electron Microscopy ◽  
Monoclonal Antibody ◽  
Human Skin ◽  
Basement Membranes ◽  
Primary Antibody ◽  
Fat Cells ◽  
Secondary Antibody ◽  
Type Vi Collagen ◽  
Dermal Matrix ◽  
Immuno Electron Microscopy

A mouse monoclonal antibody (5C6) prepared against human type VI collagen (1) has been used in this study to immunolocalize type VI collagen in human skin. The enbloc method used involves exposing whole tissue pieces to primary antibody and 5 nm gold conjugated secondary antibody before fixation, and has been described in detail elsewhere (2).Biopsies were taken from individuals ranging in age from neonate to 65 years old. By immuno-electron microscopy, type VI collagen is found to be distributed as a fine branching network closely associated with (but not attached to) banded collagen fibrils containing types I and III collagen (Fig. 1). It appears to enwrap fibers, to weave between individual fibrils within a fiber, and to span the distance separating fibers, creating a “web-like network” which entraps fibers within deep papillary and reticular dermal layers (Fig. 2). Relative to that in the dermal matrix, the concentration of type VI collagen is higher around endothelial basement membranes limiting the outer boundaries of nerves, capillaries, and fat cells (Fig. 3).

Sign in / Sign up

Export Citation Format

Share Document