scholarly journals Immunocytochemical discovery of the 22- to 23-Kd subunit of cytochrome b558 at the surface of human peripheral phagocytes

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1550-1552 ◽  
Author(s):  
M Nakamura ◽  
S Sendo ◽  
R van Zwieten ◽  
T Koga ◽  
D Roos ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Outer Surface ◽  
Small Subunit ◽  
Human Neutrophils ◽  
Cytochrome B558

Abstract A monoclonal antibody raised against cytochrome b558 reacted specifically with the 22- to 23-Kd protein, the small subunit of this cytochrome. Cytochemical studies showed that the epitope was located on the surfaces of human neutrophils and monocytes. The small subunit of cytochrome b558, therefore, was expressed at least in part on the outer surface of these cells.

scholarly journals Immunocytochemical discovery of the 22- to 23-Kd subunit of cytochrome b558 at the surface of human peripheral phagocytes

Blood ◽  
1988 ◽  
Vol 72 (5) ◽  
pp. 1550-1552
Author(s):  
M Nakamura ◽  
S Sendo ◽  
R van Zwieten ◽  
T Koga ◽  
D Roos ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Outer Surface ◽  
Small Subunit ◽  
Human Neutrophils ◽  
Cytochrome B558

A monoclonal antibody raised against cytochrome b558 reacted specifically with the 22- to 23-Kd protein, the small subunit of this cytochrome. Cytochemical studies showed that the epitope was located on the surfaces of human neutrophils and monocytes. The small subunit of cytochrome b558, therefore, was expressed at least in part on the outer surface of these cells.

scholarly journals Purification and some properties of the small subunit of cytochrome b558 from human neutrophils

1989 ◽  
Vol 264 (1) ◽  
pp. 112-118
Author(s):  
T Yamaguchi ◽  
T Hayakawa ◽  
M Kaneda ◽  
K Kakinuma ◽  
A Yoshikawa
Keyword(s):  
Small Subunit ◽  
Human Neutrophils ◽  
Cytochrome B558

scholarly journals Monoclonal antibody AHN-1 inhibits phagocytosis by human neutrophils

Blood ◽  
1985 ◽  
Vol 65 (2) ◽  
pp. 333-339
Author(s):  
KM Skubitz ◽  
DJ Weisdorf ◽  
PK Peterson
Keyword(s):  
Monoclonal Antibody ◽  
Lysosomal Enzyme ◽  
Surface Proteins ◽  
Superoxide Production ◽  
Human Neutrophils ◽  
Enzyme Release ◽  
Specific Monoclonal Antibody ◽  
Normal Human ◽  
Major Surface

The granulocyte-specific monoclonal antibody, AHN-1, immunoprecipitates two major surface-iodinated proteins of 105,000 and 145,000 to 150,000 daltons from normal human neutrophils. In this study, the effect of AHN- 1 on a number of neutrophil functions was evaluated in vitro. Both complement- and antibody-mediated phagocytosis were inhibited when human neutrophils were pretreated with AHN-1 and opsonized bacteria were used as targets. The inhibition of phagocytosis was specific, in that lysosomal enzyme release and chemotaxis were not altered by treatment with AHN-1. AHN-1 did inhibit superoxide production by neutrophils in response to particulate stimuli, but not in response to the soluble stimulus, 12-O-tetradecanoylphorbol-13-acetate. The data indicate that one or both of these surface proteins may be important in the process of phagocytosis. AHN-1 should be useful in isolating and further characterizing the nature of these molecules.

Beta 1-integrin is a maternal protein that is inserted into all newly formed plasma membranes during early Xenopus embryogenesis

Development ◽  
1992 ◽  
Vol 115 (2) ◽  
pp. 595-605 ◽  
Author(s):  
V. Gawantka ◽  
H. Ellinger-Ziegelbauer ◽  
P. Hausen
Keyword(s):  
Monoclonal Antibody ◽  
Outer Surface ◽  
Unknown Function ◽  
Plasma Membranes ◽  
Integrin Subunit ◽  
Membrane Domains ◽  
Mature Form ◽  
Maternal Mrna ◽  
Beta 1 Integrin ◽  
Early Gastrula

A monoclonal antibody (mAb 8C8) that recognizes the Xenopus beta 1-integrin chain was used to study the appearance, synthesis and distribution of this integrin subunit during the early development of Xenopus. Both the precursor and the mature form of beta 1-integrin are provided maternally. They do not increase significantly in amount until early gastrula when the level of both forms begins to rise gradually. Synthesis of beta 1-integrin from maternal mRNA is observed throughout the pregastrula phase, though it seems to add only little to the total beta 1-integrin of the embryo. Until late blastula only small amounts of precursor are processed into the mature form. Starting with the formation of the first cleavage membrane, mature beta 1-integrin is inserted into the newly formed plasma membranes of all cells. The membrane domains forming the outer surface of the embryo remain devoid of the antigen. The data suggest an as yet unknown function of beta 1-integrin during the cleavage phase.

scholarly journals Characterization of two monoclonal antibodies against cytochrome b558 of human neutrophils

Blood ◽  
1989 ◽  
Vol 73 (6) ◽  
pp. 1686-1694 ◽  
Author(s):  
AJ Verhoeven ◽  
BG Bolscher ◽  
LJ Meerhof ◽  
R van Zwieten ◽  
J Keijer ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Transmembrane Protein ◽  
Autosomal Gene ◽  
Human Neutrophils ◽  
Western Blots ◽  
Binding Characteristics ◽  
Cytochrome B558 ◽  
Spectrophotometric Methods ◽  
Membrane Bound

Abstract Monoclonal antibodies (MoAbs) were raised against cytochrome b558, a membrane-bound component of the NADPH:O2 oxidoreductase in human neutrophils. This cytochrome consists of a low-molecular-weight (low- mol-wt) subunit of 22 to 23 Kd, probably encoded by an autosomal gene, and a high-mol-wt subunit of 75 to 90 Kd, encoded on the X-chromosome. MoAb 449 reacts with the low-mol-wt subunit and MoAb 48 with the high- mol-wt subunit on Western blots of purified cytochrome b558 and on blots of whole neutrophil extracts. In extracts of neutrophils from patients with chronic granulomatous disease (CGD) in which cytochrome b558 is not detectable by spectrophotometric methods, the low-mol-wt subunit is present, albeit in a much smaller amount. The high-mol-wt subunit is not detected by MoAb 48 in neutrophils of patients with X- linked CGD and in neutrophils of patients with the autosomal, cytochrome-b558-negative form of the disease. These results can be explained by a marked instability of these subunits when the synthesis of either of the two is disturbed. In differentiated HL-60 cells, the high-mol-wt subunit appears to be present in a different form. Cloning of the low-mol-wt subunit with the help of MoAb 449 suggests the presence of a heme-binding site on this subunit. By comparison of the binding characteristics of MoAb 449 to intact and permeabilized neutrophils with those of MoAb 7D5, recently isolated by Nakamura et al (Blood 69:1404, 1987), the low-mol-wt subunit was established as a transmembrane protein.

scholarly journals Correlation between the stimulation of human neutrophil function by monoclonal antibody and by colony-stimulating factor

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 738-741 ◽  
Author(s):  
MA Vadas ◽  
C Clarke ◽  
NA Nicola ◽  
AF Lopez
Keyword(s):  
Monoclonal Antibody ◽  
Neutrophil Function ◽  
Common Mechanism ◽  
Human Neutrophils ◽  
Target Cells ◽  
Colony Stimulating Factor ◽  
Positive Correlation ◽  
Stimulating Factor ◽  
Stimulation Of

Abstract Purified human neutrophils from 48 individuals were tested for their capacity to kill antibody-coated target cells in vitro in the absence or presence of stimulating agents. The agents used to stimulate cytotoxic capacity were the monoclonal antibody (MAb) WEM-G1, colony- stimulating factor (CSF-alpha), or mononuclear cell supernatant (MNC- SN). There existed an heterogeneity among the neutrophils of different individuals in the capacity to kill target cells both in the unstimulated (“resting”) or the stimulated state. A positive correlation was found between the ability of neutrophils to kill in the “resting” state and their capacity to be stimulated by MAb WEM-G1, CSF- alpha, or MNC-SN. Furthermore, a strong positive correlation in the ability of neutrophils to be stimulated by the MAb WEM-G1 and either CSF-alpha (r = .76) or MNC-SN (r = .68), as well as between CSF-alpha and MNC-SN (r = .79) was demonstrated. No correlation was seen, however, between stimulation of neutrophil function in vitro and total blood leukocyte counts, neutrophil counts, monocyte counts, or intensity of binding of MAb WEM-G1. The observation that neutrophils respond to a similar extent to different types of stimulators, -such as cytokines (CSF-alpha and MNC-SN) and MAb, suggests that these two factors may be operating through a common mechanism and the degree of stimulation may reflect an intrinsic responsiveness of neutrophils that differs among individuals. Our results also suggest a potential clinical use of WEM-G1 in measuring neutrophil functional capacity in vitro and predicting the capacity to respond to CSF-like cytokines.

scholarly journals Thrombospondin stimulates motility of human neutrophils.

1990 ◽  
Vol 111 (6) ◽  
pp. 3077-3086 ◽  
Author(s):  
P J Mansfield ◽  
L A Boxer ◽  
S J Suchard
Keyword(s):  
Monoclonal Antibody ◽  
Cell Types ◽  
Human Neutrophils ◽  
Heparin Binding ◽  
Directed Movement ◽  
Surface Receptors ◽  
Chemotactic Peptides ◽  
Heparin Binding Domain ◽  
High Concentrations ◽  
Priming Activity

Polymorphonuclear leukocytes (PMNs) migrate to sites of inflammation or injury in response to chemoattractants released at those sites. The presence of extracellular matrix (ECM) proteins at these sites may influence PMN accumulation at blood vessel walls and enhance their ability to move through tissue. Thrombospondin (TSP), a 450-kD ECM protein whose major proteolytic fragments are a COOH-terminal 140-kD fragment and an NH2-terminal heparin-binding domain (HBD), is secreted by platelets, endothelial cells, and smooth muscle cells. TSP binds specifically to PMN surface receptors and has been shown, in other cell types, to promote directed movement. TSP in solution at low concentrations (30-50 nM) "primed" PMNs for f-Met-Leu-Phe (fMLP)-mediated chemotaxis, increasing the response two- to fourfold. A monoclonal antibody against the HBD of TSP totally abolished this priming effect suggesting that the priming activity resides in the HBD of TSP. Purified HBD retains the priming activity of TSP thereby corroborating the antibody data. TSP alone, in solution at high concentrations (0.5-3.0 microM), stimulated chemotaxis of PMNs and required both the HBD and the 140-kD fragment of TSP. In contrast to TSP in solution, TSP bound to nitrocellulose filters in the range of 20-70 pmol stimulated random locomotion of PMNs. The number of PMNs migrating in response to bound TSP was approximately two orders of magnitude greater than the number of cells that exhibited chemotaxis in response to soluble TSP or fMLP. Monoclonal antibody C6.7, which recognizes an epitope near the carboxyl terminus of TSP, blocked migration stimulated by bound TSP, suggesting that the activity resides in this domain. Using proteolytic fragments, we demonstrated that bound 140-kD fragment, but not HBD, promoted migration of PMNs. Therefore, TSP released at injury sites, alone or in synergy with chemotactic peptides like fMLP, could play a role in directing PMN movement.

Fc receptors for IgG on human neutrophils: analysis of structure and function by using monoclonal antibody probes.

1985 ◽  
Vol 31 (9) ◽  
pp. 1444-1448 ◽  
Author(s):  
J S Rosenberg ◽  
M J Melnicoff ◽  
P Wilding
Keyword(s):  
Monoclonal Antibody ◽  
Fc Receptors ◽  
Myelogenous Leukemia ◽  
Human Neutrophils ◽  
Fc Gamma Receptor ◽  
Immunogold Staining ◽  
Cell Functions ◽  
Gamma Receptor ◽  
Functional Aspects ◽  
And Function

Abstract Structural and functional characteristics of Fc receptors for IgG (Fc gamma) on human neutrophils were examined with two monoclonal antibody probes specific for the Fc gamma receptors, Leu 11b and 3G8. To determine the distribution, density, and membrane mobility of the Fc gamma receptor, we used immunogold staining techniques, flow cytometry analysis, and fluorescence microscopy. Both 3G8 and Leu 11b inhibited several cell functions, thereby depicting the regulatory role of the Fc gamma receptor in mediating neutrophil activities. Among the functions studied were release of lysosomal enzymes, release of superoxide anion (O2-), and Fc-dependent rosette formation and phagocytosis. The densities of Fc gamma determinants recognized by Leu 11b and 3G8 on cells from a patient with chronic myelogenous leukemia were less than the density of epitopes on neutrophils from a normal individual. Taken together, the detailed analysis of physical and functional aspects of the Fc gamma receptor on neutrophils described in this study serve as a model for further assessment of the use of Fc gamma phenotyping of cells as a diagnostic tool.

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