Polymorphic glycoprotein-1 on mouse platelets: possible role of Pgp-1 and LFA-1 in antibody-dependent platelet cytotoxicity involving complement

The presence of the Pgp-1 glycoprotein on mouse platelets is demonstrated by antibody-binding techniques, by immunoprecipitation, and by transblotting using the monoclonal antibody (MoAb) C71/26 against Pgp-1. C71/26 immunoprecipitates as a broad band of mol wt 87,000 to 100,000 as determined by radioiodination of the platelet cell surface and by the 3H-sodium borohydride labeling technique. Immunoblotting showed Pgp-1 expression on platelets to be quantitatively similar to its presence on macrophages and resolved platelet Pgp-1 into two bands of mol wt 87,000 and 97,000 whereas Pgp-1 on parasite-elicited peritoneal macrophages showed 82,000 and 87,000 mol wt species. Platelets and monocyte/macrophage cells from either peripheral blood or from the peritoneal cavity showed homogeneous binding of Pgp-1 antibody to greater than 97% of cells by flow cytometry. In contrast, lymphocytes from peripheral blood or from the spleen showed a heterogeneous binding pattern with 20% to 30% of cells being negative, and the majority weakly positive. In functional studies, MoAbs against CR1 and CR3 substantially inhibited platelet immune adherence, whereas C71/26 showed only marginal inhibitor. In contrast, C71/26 and other MoAbs against Pgp-1 inhibited platelet- dependent cytotoxicity of antibody-coated sheep erythrocytes in the presence of C5-deficient mouse plasma whereas M1/70 against CR3 showed no effect. In this assay, MoAbs against the alpha- and beta-subunits of leukocyte functional molecule LFA-1 also inhibited platelet cytotoxicity. These results show that the platelet cell surface moieties Pgp-1 and LFA-1 are involved in or closely associated with antibody-dependent cellular cytotoxicity by platelets.

Download Full-text

Polymorphic glycoprotein-1 on mouse platelets: possible role of Pgp-1 and LFA-1 in antibody-dependent platelet cytotoxicity involving complement

Blood ◽

10.1182/blood.v69.1.211.211 ◽

1987 ◽

Vol 69 (1) ◽

pp. 211-218 ◽

Cited By ~ 1

Author(s):

PJ McCaffery ◽

AS Tan ◽

MV Berridge

Keyword(s):

Cell Surface ◽

Peripheral Blood ◽

Broad Band ◽

Peritoneal Macrophages ◽

Beta Subunits ◽

Deficient Mouse ◽

Antibody Dependent Cellular Cytotoxicity ◽

Functional Studies ◽

Immune Adherence

Abstract The presence of the Pgp-1 glycoprotein on mouse platelets is demonstrated by antibody-binding techniques, by immunoprecipitation, and by transblotting using the monoclonal antibody (MoAb) C71/26 against Pgp-1. C71/26 immunoprecipitates as a broad band of mol wt 87,000 to 100,000 as determined by radioiodination of the platelet cell surface and by the 3H-sodium borohydride labeling technique. Immunoblotting showed Pgp-1 expression on platelets to be quantitatively similar to its presence on macrophages and resolved platelet Pgp-1 into two bands of mol wt 87,000 and 97,000 whereas Pgp-1 on parasite-elicited peritoneal macrophages showed 82,000 and 87,000 mol wt species. Platelets and monocyte/macrophage cells from either peripheral blood or from the peritoneal cavity showed homogeneous binding of Pgp-1 antibody to greater than 97% of cells by flow cytometry. In contrast, lymphocytes from peripheral blood or from the spleen showed a heterogeneous binding pattern with 20% to 30% of cells being negative, and the majority weakly positive. In functional studies, MoAbs against CR1 and CR3 substantially inhibited platelet immune adherence, whereas C71/26 showed only marginal inhibitor. In contrast, C71/26 and other MoAbs against Pgp-1 inhibited platelet- dependent cytotoxicity of antibody-coated sheep erythrocytes in the presence of C5-deficient mouse plasma whereas M1/70 against CR3 showed no effect. In this assay, MoAbs against the alpha- and beta-subunits of leukocyte functional molecule LFA-1 also inhibited platelet cytotoxicity. These results show that the platelet cell surface moieties Pgp-1 and LFA-1 are involved in or closely associated with antibody-dependent cellular cytotoxicity by platelets.

Download Full-text

The Role of Sialic Acids in the Establishment of Infections by Pathogens, With Special Focus on Leishmania

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.671913 ◽

2021 ◽

Vol 11 ◽

Author(s):

Tainá Cavalcante ◽

Mariana Medina Medeiros ◽

Simon Ngao Mule ◽

Giuseppe Palmisano ◽

Beatriz Simonsen Stolf

Keyword(s):

Sialic Acid ◽

Cell Surface ◽

Host Cell ◽

Functional Diversity ◽

Sialic Acids ◽

Cellular Communication ◽

Special Focus ◽

Functional Studies ◽

Sialic Acid Binding

Carbohydrates or glycans are ubiquitous components of the cell surface which play crucial biological and structural roles. Sialic acids (Sias) are nine-carbon atoms sugars usually present as terminal residues of glycoproteins and glycolipids on the cell surface or secreted. They have important roles in cellular communication and also in infection and survival of pathogens. More than 20 pathogens can synthesize or capture Sias from their hosts and incorporate them into their own glycoconjugates and derivatives. Sialylation of pathogens’ glycoconjugates may be crucial for survival inside the host for numerous reasons. The role of Sias in protozoa such as Trypanosoma and Leishmania was demonstrated in previous studies. This review highlights the importance of Sias in several pathogenic infections, focusing on Leishmania. We describe in detail the contributions of Sias, Siglecs (sialic acid binding Ig-like lectins) and Neuraminidase 1 (NEU 1) in the course of Leishmania infection. A detailed view on the structural and functional diversity of Leishmania-related Sias and host-cell receptors will be provided, as well as the results of functional studies performed with different Leishmania species.

Download Full-text

Inhibition of N-glycosylation affects transepithelial Na+ but not Na+-K+-ATPase transport

AJP Cell Physiology ◽

10.1152/ajpcell.1989.256.5.c958 ◽

1989 ◽

Vol 256 (5) ◽

pp. C958-C966 ◽

Cited By ~ 21

Author(s):

D. Zamofing ◽

B. C. Rossier ◽

K. Geering

Keyword(s):

Cell Surface ◽

Short Circuit ◽

Short Circuit Current ◽

Beta Subunit ◽

Beta Subunits ◽

Toad Urinary Bladder ◽

Na Transport ◽

Atpase Inhibition ◽

Dose Dependent

Tunicamycin (TM) was used in toad urinary bladder (TBM) cells to study the role of N-glycosylation of the beta-subunit of Na+-K+-ATPase. Inhibition of the beta-subunit core glycosylation was dose dependent and coincided with a specific 70% decrease in newly synthesized beta- and alpha-subunits. Na+-K+-ATPase activity paralleled the decrease in the cellular content of the alpha-subunit, although the cellular and cell surface-expressed Na+-K+-ATPase pool was progressively filled up with nonglycosylated beta-subunits. In addition, the decrease in maximal Na+ transport capacity of the Na+-K+-ATPase as assessed by short-circuit current (SCC) measurements in the presence of amphotericin B correlated with the decrease in the total cell surface-expressed beta-subunit population despite the fact that it was composed of 47% nonglycosylated beta-subunits after 42 h of TM treatment. These results are consistent with the interpretation that beta-subunit glycosylation is not important either for the enzyme's intracellular sorting to the plasma membrane or its hydrolytic and transport properties. Finally, TM produced effects on basal SCC and electrical resistance that differed in their times of onset and time periods needed for recovery. Thus, in addition to the Na+-K+-ATPase, other glycoproteins in the apical membrane and the tight junctions must be implicated in the maintenance of transepithelial Na+ transport.

Download Full-text

New insights suggesting a possible role of a heat shock protein 70-kD family-related protein in antigen processing/presentation phenomenon in humans

Blood ◽

10.1182/blood.v82.9.2865.2865 ◽

1993 ◽

Vol 82 (9) ◽

pp. 2865-2871 ◽

Cited By ~ 32

Author(s):

GC Manara ◽

P Sansoni ◽

L Badiali-De Giorgi ◽

G Gallinella ◽

C Ferrari ◽

...

Keyword(s):

Cell Surface ◽

Antigen Processing ◽

Potential Role ◽

Related Protein ◽

Functional Studies ◽

Recall Antigen ◽

Normal Human ◽

Antigen Presenting ◽

Antigen Processing And Presentation

Abstract A possible role of the peptide binding protein (PBP) 72/74 in antigen processing and presentation has been recently suggested in mice. In order to evaluate a possible analogous role of a PBP72/74-related protein in humans, immunoelectron microscope investigations, functional studies, and immunofluorescence analyses were performed on normal human peripheral antigen-presenting cells. We demonstrated that the determinant recognized by antiheat shock protein (HSP) 72/73 monoclonal antibody (MoAb) is constitutively expressed on the cell surface of monocytes as well as of B cells. Moreover, the capability of monocytes to present a recall antigen to T cells was significantly decreased when preincubated with an anti-HSP72/73 MoAb. These data add further strength to a potential role of a protein related to human PBP72/74 homologue in antigen processing and/or presentation. Finally, the capability of anti-HSP72/73 MoAb to impair the ability of fixed monocytes to present a synthetic peptide demonstrates that cell surface- localized PBP72/74-related protein could play a role in antigen presentation.

Download Full-text

Role of Acetylsalicylic Acid in Cytokine Stimulation of Macrophages in Antibody-Dependent Cellular Cytotoxicity (ADCC)

Mediators of Inflammation ◽

10.1155/s0962935194000591 ◽

1994 ◽

Vol 3 (6) ◽

pp. 419-424 ◽

Cited By ~ 1

Author(s):

M. Jäpel ◽

H. Lötzerich ◽

K. Rogalla

Keyword(s):

Acetylsalicylic Acid ◽

Binding Capacity ◽

Interleukin 2 ◽

Peritoneal Macrophages ◽

Biological Action ◽

Interleukin 4 ◽

Cellular Cytotoxicity ◽

Antibody Dependent Cellular Cytotoxicity ◽

Cytokine Stimulation

In addition to the spectrum of biological action already known to be exhibited by acetylsalicylic acid (ASA) as an analgesic, anti-inflammatory and platelet aggregation inhibitor, there is growing evidence of a stimulatory effect on the immune system. ASA has been found to increase the production ofcytokines and to increase the activity of various leukocytes. The action of ASA on the activity of mouse peritoneal macrophages was therefore investigated in the present study. Therapeutically effective concentrations of ASA, which are known to decrease levels of prostaglandins, had neither a stimulating nor an inhibiting influence on antibody-dependent cellular cytotoxicity (ADCC) or on the binding capacity of macrophages with regard to SW 948 tumour cells. Likewise ASA had little or no adverse effect on the capacity of the macrophages for stimulation by interferon-gamma (IFN-gamma) and interleukin-4 (IL-4). Taken together, the immunostimulant effect of ASA shown in the literature as an increased production of interleukin-2 (IL-2) and IFN, could not be confirmed on the basis of the macrophage cytotoxiclty.

Download Full-text

New insights suggesting a possible role of a heat shock protein 70-kD family-related protein in antigen processing/presentation phenomenon in humans

Blood ◽

10.1182/blood.v82.9.2865.bloodjournal8292865 ◽

1993 ◽

Vol 82 (9) ◽

pp. 2865-2871

Author(s):

GC Manara ◽

P Sansoni ◽

L Badiali-De Giorgi ◽

G Gallinella ◽

C Ferrari ◽

...

Keyword(s):

Cell Surface ◽

Antigen Processing ◽

Potential Role ◽

Related Protein ◽

Functional Studies ◽

Recall Antigen ◽

Normal Human ◽

Antigen Presenting ◽

Antigen Processing And Presentation

A possible role of the peptide binding protein (PBP) 72/74 in antigen processing and presentation has been recently suggested in mice. In order to evaluate a possible analogous role of a PBP72/74-related protein in humans, immunoelectron microscope investigations, functional studies, and immunofluorescence analyses were performed on normal human peripheral antigen-presenting cells. We demonstrated that the determinant recognized by antiheat shock protein (HSP) 72/73 monoclonal antibody (MoAb) is constitutively expressed on the cell surface of monocytes as well as of B cells. Moreover, the capability of monocytes to present a recall antigen to T cells was significantly decreased when preincubated with an anti-HSP72/73 MoAb. These data add further strength to a potential role of a protein related to human PBP72/74 homologue in antigen processing and/or presentation. Finally, the capability of anti-HSP72/73 MoAb to impair the ability of fixed monocytes to present a synthetic peptide demonstrates that cell surface- localized PBP72/74-related protein could play a role in antigen presentation.

Download Full-text

Role of the transmembrane and extracytoplasmic domain of beta subunits in subunit assembly, intracellular transport, and functional expression of Na,K-pumps.

The Journal of Cell Biology ◽

10.1083/jcb.123.6.1751 ◽

1993 ◽

Vol 123 (6) ◽

pp. 1751-1759 ◽

Cited By ~ 58

Author(s):

P Jaunin ◽

F Jaisser ◽

A T Beggah ◽

K Takeyasu ◽

P Mangeat ◽

...

Keyword(s):

Cell Surface ◽

Intracellular Transport ◽

Transmembrane Domain ◽

Functional Expression ◽

Beta Subunit ◽

Beta Subunits ◽

Chimeric Proteins ◽

Er Retention ◽

Retention Signal

The ubiquitous Na,K- and the gastric H,K-pumps are heterodimeric plasma membrane proteins composed of an alpha and a beta subunit. The H,K-ATPase beta subunit (beta HK) can partially act as a surrogate for the Na,K-ATPase beta subunit (beta NK) in the formation of functional Na,K-pumps (Horisberger et al., 1991. J. Biol. Chem. 257:10338-10343). We have examined the role of the transmembrane and/or the ectodomain of beta NK in (a) its ER retention in the absence of concomitant synthesis of Na,K-ATPase alpha subunits (alpha NK) and (b) the functional expression of Na,K-pumps at the cell surface and their activation by external K+. We have constructed chimeric proteins between Xenopus beta NK and rabbit beta HK by exchanging their NH2-terminal plus transmembrane domain with their COOH-terminal ectodomain (beta NK/HK, beta HK/NK). We have expressed these constructs with or without coexpression of alpha NK in the Xenopus oocyte. In the absence of alpha NK, Xenopus beta NK and all chimera that contained the ectodomain of beta NK were retained in the ER while beta HK and all chimera with the ectodomain of beta HK could leave the ER suggesting that ER retention of unassembled Xenopus beta NK is mediated by a retention signal in the ectodomain. When coexpressed with alpha NK, only beta NK and beta NK/HK chimera assembled efficiently with alpha NK leading to similar high expression of functional Na,K-pumps at the cell surface that exhibited, however, a different apparent K+ affinity. beta HK or chimera with the transmembrane domain of beta HK assembled less efficiently with alpha NK leading to lower expression of functional Na,K-pumps with a different apparent K+ affinity. The data indicate that the transmembrane domain of beta NK is important for efficient assembly with alpha NK and that both the transmembrane and the ectodomain of beta subunits play a role in modulating the transport activity of Na,K-pumps.

Download Full-text

Ionic homeostasis and subcellular element compartmentation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010013434x ◽

1990 ◽

Vol 48 (2) ◽

pp. 150-151

Author(s):

Ann LeFurgey ◽

Peter Ingram ◽

J.J. Blum ◽

M.C. Carney ◽

L.A. Hawkey ◽

...

Keyword(s):

Leishmania Major ◽

Ionic Homeostasis ◽

X Ray ◽

Functional Studies ◽

Cell Compartments ◽

X Ray Imaging ◽

Resolution Electron ◽

Multiple Cell ◽

Role Of Zn

Subcellular compartments commonly identified and analyzed by high resolution electron probe x-ray microanalysis (EPXMA) include mitochondria, cytoplasm and endoplasmic or sarcoplasmic reticulum. These organelles and cell regions are of primary importance in regulation of cell ionic homeostasis. Correlative structural-functional studies, based on the static probe method of EPXMA combined with biochemical and electrophysiological techniques, have focused on the role of these organelles, for example, in maintaining cell calcium homeostasis or in control of excitation-contraction coupling. New methods of real time quantitative x-ray imaging permit simultaneous examination of multiple cell compartments, especially those areas for which both membrane transport properties and element content are less well defined, e.g. nuclei including euchromatin and heterochromatin, lysosomes, mucous granules, storage vacuoles, microvilli. Investigations currently in progress have examined the role of Zn-containing polyphosphate vacuoles in the metabolism of Leishmania major, the distribution of Na, K, S and other elements during anoxia in kidney cell nuclel and lysosomes; the content and distribution of S and Ca in mucous granules of cystic fibrosis (CF) nasal epithelia; the uptake of cationic probes by mltochondria in cultured heart ceils; and the junctional sarcoplasmic retlculum (JSR) in frog skeletal muscle.

Download Full-text

The Role of the Interaction between Fe(III) and Cell Surface in the Accumulation of 67Ga by Tumor Cells

Nuklearmedizin ◽

10.1055/s-0038-1629590 ◽

1991 ◽

Vol 30 (06) ◽

pp. 290-293 ◽

Cited By ~ 1

Author(s):

P. Maleki ◽

A. Martinezi ◽

M. C. Crone-Escanye ◽

J. Robert ◽

L. J. Anghileri

Keyword(s):

Cell Surface ◽

Tumor Cells ◽

Ionic Composition ◽

Iron Complex ◽

Iron Binding ◽

Complex Time ◽

Interaction Product ◽

Thermodynamic Constant ◽

Number Of Cells

The study of the interaction between complexed iron and tumor cells in the presence of 67Ga-citrate indicates that a phenomenon of iron-binding related to the thermodynamic constant of stability of the iron complex, and a hydrolysis (or anion penetration) of the interaction product determine the uptake of 67Ga. The effects of various parameters such as ionic composition of the medium, nature of the iron complex, time of incubation and number of cells are discussed.

Download Full-text

The Local Sanarelli-Shwartzman Phenomenon in Rats Induced by Human Leukaemic Cells

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647777 ◽

1973 ◽

Vol 29 (02) ◽

pp. 353-362

Author(s):

J Lisiewicz ◽

A Pituch ◽

J. A Litwin

Keyword(s):

Chronic Lymphocytic Leukaemia ◽

Intravenous Injection ◽

Peripheral Blood ◽

Lymphocytic Leukaemia ◽

Acute Myeloblastic Leukaemia ◽

Demarcation Line ◽

E Coli ◽

Leukaemic Cells ◽

Shwartzman Phenomenon

SummaryThe local Sanarelli-Shwartzman phenomenon (SSP-L) in the skin of 30 rats was induced by an intr a cutaneous sensitizing injection of leukaemic leucocytes isolated from the peripheral blood of patients with chronic lymphocytic leukaemia (CLL), acute myeloblastic leukaemia (AL) and chronic granulocytic leukaemia (CGL) and challenged by an intravenous injection of 100(μ of E. coli endotoxin. SSP-L was observed in 7 rats after injection of CLL lymphocytes and in 6 and 2 rats after AL myeloblasts and the CGL granulocytes, respectively. The lesions in the skin after AL myeloblasts appeared in a shorter time and were of longer duration compared with those observed after CLL lymphocytes and CGL granulocytes. Histologically, the lesions consisted of areas of destruction in the superficial layers of the skin ; the demarcation line showed the presence of neutrophils, macrophages and erythrocytes. Haemorrhages and fibrin deposits near the demarcation line were larger after injection of CLL lymphocytes and AL myeloblasts than after CGL granulocytes. The possible role of leucocyte procoagulative substances in the differences observed have been discussed.

Download Full-text