Antibodies to porcine uteroferrin used in measurement of human tartrate-resistant acid phosphatase.

Echetebu, Z. O.; Cox, Timothy M.; Moss, D. W.

doi:10.1093/clinchem/33.10.1832

Cited by 34 publications

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“…These cells have a common lineage from myeloid progenitor cells and appear to be immunologically distinct from Langerhans' and splenic dendritic cells that are non-classical Mw. 20 Immunostaining of Mw isolated from both sources using immune-speci®c polyclonal antiuteroferrin antibody showed abundant intracellular staining attributable to TRAP in cells obtained from wildtype animals; 15 as expected, immunostaining was absent in cells from Acp 5 null mice lacking TRAP enzymatic activity ( Fig. 1b, 1c).…”

Section: Immunophenotype Of Trap-de®cient and Wild-type Peritoneal Anmentioning

confidence: 89%

Mice lacking tartrate‐resistant acid phosphatase (Acp 5) have disordered macrophage inflammatory responses and reduced clearance of the pathogen, Staphylococcus aureus

et al. 2001

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SUMMARYTartrate-resistant acid phosphatase (TRAP) is a lysosomal di-iron protein of mononuclear phagocytes and osteoclasts. Hitherto, no role for the enzyme in immunity has been identi®ed; however, knockout mice lacking TRAP have a skeletal phenotype caused by an intrinsic osteoclast defect. To investigate a putative function for TRAP in macrophages (Mw), we investigated proin¯ammatory responses and systemic microbial clearance in knockout mice compared with age-and gendermatched congenic wild-type mice. Phorbol 12-myristate 13-acetate (PMA)-stimulated and interferon-c (IFN-c)-induced superoxide formation was enhanced in peritoneal Mw lacking TRAP; nitrite production in response to stimulation with lipopolysaccharide (LPS) and IFN-c was also increased. In addition, secretion of the proin¯ammatory cytokines, tumour necrosis factor-a (TNF-a), interleukin (IL)-1b and IL-12, was signi®cantly greater in TRAP-de®cient Mw when stimulated with LPS, with or without addition of either TNF-a or IFN-c. The activity of tartratesensitive (lysosomal) acid phosphatase was increased in Mw from the knockout mice but activities of the lysosomal hydrolases N-acetyl b-glucosaminidase and acid b-glucuronidase were unchanged, indicating selective activation of compensatory acid phosphatase activity. Evidence of impaired Mw function in vivo was obtained in TRAP knockout mice, which showed delayed clearance of the microbial pathogen, Staphylococcus aureus, after sublethal intraperitoneal inoculation. After microbial challenge, peritoneal exudates obtained from TRAP knockout mice had a reduced population of Mw. As peritoneal Mw and neutrophils lacking TRAP were able to phagocytose and kill S. aureus normally in vitro, TRAP may directly or indirectly in¯uence recruitment of Mw to sites of microbial invasion. Our study shows that TRAP participates in the in¯ammatory response of the Mw and in¯uences effector signalling pathways in innate immunity.

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Section: Immunophenotype Of Trap-de®cient and Wild-type Peritoneal Anmentioning

confidence: 89%

Mice lacking tartrate‐resistant acid phosphatase (Acp 5) have disordered macrophage inflammatory responses and reduced clearance of the pathogen, Staphylococcus aureus

et al. 2001

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“…The specificity of the antibody to acid phosphatase has been further confirmed by electrophoresis of an unfractionated extract of Gaucher's disease spleen on SDS(sodium dodecyl sulphate)-polyacrylamide gel followed by probing with anti-uteroferrin antiserum and 1251-labelled protein A. This has shown that the antiserum reacts only with the 37 kDa protein band corresponding to the iron-containing acid phosphatase isoenzyme and with no other protein (Echetebu et al 1987). The cross-reactivity of anti-porcine uteroferrin antibody with rat osteoclastic acid phosphatase was confirmed in the following way.…”

Section: Measurement Of Acid Phosphatase Activitymentioning

confidence: 90%

“…The antibody-isoenzyme complex has been shown to possess markedly reduced catalytic activity. The antiserum or immunopurified antibodies also have been found not to cross-react with other acid phosphatase isoenzymes, including the tartrate-resistant erythrocyte enzyme (Echetebu et al 1987). The specificity of the antibody to acid phosphatase has been further confirmed by electrophoresis of an unfractionated extract of Gaucher's disease spleen on SDS(sodium dodecyl sulphate)-polyacrylamide gel followed by probing with anti-uteroferrin antiserum and 1251-labelled protein A.…”

Section: Measurement Of Acid Phosphatase Activitymentioning

confidence: 91%

“…Anti-uteroferrin antiserum An antiserum against purified porcine uteroferrin (an iron-containing protein with acid phosphatase activity) had been raised in rabbit and anti-uteroferrin IgG antibodies purified by specific immunoaffinity chromatography as described previously (Echetebu, Cox & Moss, 1987). The antiserum had been shown to cross-react completely with human tartrate-resistant acid phosphatase from Gaucher's disease spleen and alveolar macrophages, and with the increased acid phosphatase activity in sera of patients with osteoclastic bone diseases.…”

Section: Measurement Of Acid Phosphatase Activitymentioning

confidence: 99%

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Intracellular regulation of enzyme secretion from rat osteoclasts and evidence for a functional role in bone resorption.

Moonga

Moss

Patchell

et al. 1990

The Journal of Physiology

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115

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SUMMARY1. Osteoclasts are known to secrete acid phosphatase, an iron-containing phosphohydrolase. We have investigated (a) the possibility that acid phosphatase has a functional role in bone resorption and (b) the factors controlling enzyme secretion from isolated rat osteoclasts.2. Osteoclasts were freshly disaggregated from neonatal rat long bones and dispersed at low densities on devitalized cortical bone slices or on plastic substrate. The levels of acid phosphatase in culture medium were measured spectrophotometrically using 4-nitrophenyl phosphate as hydrolysable substrate. The total plan area of bone resorbed was quantified by scanning electron microscopy in combination with image processing and analysis.3. Ninety-three per cent of the total enzyme activity detected in the supernatant exposed to bone-osteoclast preparations was resistant to inhibition by D-tartaric acid and was bound to an antibody known to be highly specific for the osteoclastderived isoenzyme, showing that it originated from osteoclasts.4. A diminution in the level of supernatant enzyme activity achieved by incubating bone-osteoclast preparations with an antiserum specifically binding the osteoclast isoenzyme, or with a non-competitive inhibitor, molybdate or with competitive inhibitors, disphosphonates, led to a marked reduction of osteoclastic bone resorption.5. The rate of the enzyme released into the culture supernatant, whether from resorbing (cultured on bone) or non-resorbing (cultured on plastic) osteoclasts declined gradually over 22 h, but that from the former was significantly depressed within the first 30 min of incubation. The supernatant enzyme concentration increased linearly up to 3 h; the levels released from resorbing osteoclasts remained consistently lower than those from non-resorbing cells. B. S. MOONGA AND OTHERS 7. Exposure of bone-osteoclast preparations to pertussis toxin produced no significant change of acid phosphatase release, while cholera toxin, dibutyryl cyclic AMP and forskolin produced a marked elevation of enzyme secretion. lonomycin was found to inhibit enzyme release and this was less marked when osteoclasts were incubated on plastic substrate.8. There was a significant positive correlation between basal enzyme release and bone resorption (correlation coefficient 0-78; P < 0-01) and between the percentage fall in supernatant enzyme activity and the percentage reduction of measured resorption in the presence of elevated [Ca2+] generated as a result of osteoclastic resorption. This suggests the existence of an important self-regulatory mechanism. (c) The intracellular regulation of acid phosphatase release is effected via the intermediacy of G proteins, one of which is cholera toxin sensitive and causes stimulation of enzyme release via a cyclic AMPdependent pathway.

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“…The anti-human cathepsin D antibody was described previously (Muschol et al 2002). The polyclonal anti-porcine uteroferrin antibody (Echetebu et al 1987) was a kind gift from Dr Cox (University of Cambridge, Cambridge, UK). Secondary horseradish peroxidase-conjugated goat anti-rabbit, goat anti-mouse, and goat anti-rat IgG were obtained from Dianova (Hamburg, Germany).…”

Section: Antibodiesmentioning

confidence: 99%

Increased expression of lysosomal acid phosphatase in CLN3‐defective cells and mouse brain tissue

Pohl

Mitchison

Kohlschütter

et al. 2007

Journal of Neurochemistry

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Juvenile neuronal ceroid lipofuscinosis (Batten disease) is a neurodegenerative disorder caused by defective function of the lysosomal membrane glycoprotein CLN3. The activity of the lysosomal acid phosphatase (LAP/ACP2) was found to be significantly increased in the cerebellum and brain stem of Cln3-targeted mice during the early stages of postnatal life. Histochemical localization studies revealed an increased LAP/ ACP2 staining intensity in neurons of the cerebral cortex of 48-week-old Cln3-targeted mice as compared with controls. Additionally, the expression of another lysosomal membrane protein LAMP-2 was increased in all brain areas. Knockdown of CLN3 expression in HeLa cells by RNA interference also resulted in increased LAP/ACP2 and LAMP-2 expression. The juvenile form of neuronal ceroid lipofuscinosis (JNCL, Batten disease) is a recessively inherited neurodegenerative disease of childhood which is characterized by initial gliosis and astrocytosis followed by the progressive loss of neurons in the cortex, hippocampus, cerebellum, and the retina . Clinical features include visual loss, mental and motor retardation, epilepsy, and premature death. JNCL belongs to a group of neurodegenerative diseases with defects in ten different genes encoding both soluble and polytopic membrane proteins located in either late endosomes/lysosomes or the endoplasmic reticulum The function of CLN3 is unknown and several processes have been proposed to involve the CLN3 protein such as import of arginine into lysosomes, maturation of autophagic vacuoles, vesicular trafficking, modulation of ceramide synthesis, and apoptosis (Persaud-Sawin et al. 2002, 2004Kim et al. 2003;Luiro et al. 2004;Cao et al. 2006). The role of CLN3 in lysosomal pH homeostasis has been a matter of debate depending on the cell type and model organism being studied reporting either an increase (Pearce Received April 5, 2007; Revised July 27, 2007; Accepted August 6, 2007. Address correspondence and reprint requests to Stephan Storch, Department of Biochemistry, Children's Hospital, University Medical Center Hamburg, D-20246 Hamburg, Germany. E-mail: storch@uke.uni-hamburg.deAbbrevations used: ACP2, human acid phosphatase 2; ACP5, human acid phosphatase 5; ACTB, human b-actin; ActB, mouse b-actin; JNCL, juvenile form of neuronal ceroid lipofuscinosis; LAMP-1, lysosomeassociated membrane protein 1; LAMP-2, lysosome-associated membrane protein 2; LAP, lysosomal acid phosphatase; M6P, mannose 6-phosphate; P week, postnatal week; RT, real-time; siRNA, small interfering RNA; TRAP, tartrate-resistant acid phosphatase.

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Antibodies to porcine uteroferrin used in measurement of human tartrate-resistant acid phosphatase.

Cited by 34 publications

References 0 publications

Mice lacking tartrate‐resistant acid phosphatase (Acp 5) have disordered macrophage inflammatory responses and reduced clearance of the pathogen, Staphylococcus aureus

Mice lacking tartrate‐resistant acid phosphatase (Acp 5) have disordered macrophage inflammatory responses and reduced clearance of the pathogen, Staphylococcus aureus

Intracellular regulation of enzyme secretion from rat osteoclasts and evidence for a functional role in bone resorption.

Increased expression of lysosomal acid phosphatase in CLN3‐defective cells and mouse brain tissue

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