RESUMEN
The serum precursor SAA of the secondary amyloid protein AA has been detected by solid-phase radioimmunoassay as a normal serum alpha-globulin of mol wt 160,000, which dissociates to a more stable 12,500 dalton moiety on treatment with formic acid. In 12 strains of mice, including T-cell-deficient nude mice, treated with the amyloid-inducing agents lipopolysaccharide (LPS) or casein, SAA behaved as an acute-phase reactant. SAA concentration rose to about 750 mug/ml by 24 h and returned to less than 1 mug/ml by 48 h. Since the amyloid-resistant colchicine-treated mice and AJ mice had a normal SAA response to LPS, it appears that their resistance to amyloid induction is due to the nature of their SAA processing rather than decreased SAA production. C3H/HeJ mice, which have defective B-lymphocyte responses to LPS, required extremely high dosages of LPS to cause SAA elevation, although their SAA response to casein was normal. This suggests that SAA is an acute-phase protein produced as a result of B-lymphocyte stimulation. Preliminary evidence suggests that at the height of an acute SAA response, liver homogenates are particularly rich in protein AA cross-reacting material.
Asunto(s)
Amiloide/metabolismo , Amiloidosis/sangre , Proteínas Sanguíneas/metabolismo , Variación Genética , Enfermedad Aguda , Animales , Caseínas , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos , Polisacáridos Bacterianos , Radioinmunoensayo , Especificidad de la EspecieRESUMEN
The mechanism by which LPS stimulates an acute phase serum amyloid A (SAA) response in C3H mice has been studied. A factor (SAA inducer) appears in the blood of C3H/HeN (lipopolysaccharide [LPS]-sensitive) mice approximately 1 h after administration of LPS, which, when passively administered, can induce C3H/HeJ mice to produce SAA although they are resistant to the LPS itself. SAA inducer has been detected in the culture medium of LPS treated C3H/HeN macrophages but not spleen cells. Thus, two stages in the induction of the acute phase SAA response are now recognized: a latent period of 2-3 h during which the SAA concentration remains at baseline values and in which SAA inducer appears, and the period of synthesis of SAA which lasts for approoximately 24 h past induction. It is proposed that a macrophage response to LPS is responsible for production of the serum mediator which induces SAA synthesis.
Asunto(s)
Amiloide/biosíntesis , Endotoxinas/farmacología , Macrófagos/fisiología , Proteína Amiloide A Sérica/biosíntesis , Animales , Quimera , Femenino , Inmunización Pasiva , Lipopolisacáridos/farmacología , Masculino , Ratones , Ratones Endogámicos C3H , Bazo/inmunología , Factores de TiempoRESUMEN
Inbred strains of mice provide a model for studies of the pathogenesis of amyloid A (AA) amyloidosis. All susceptible strains of mice described to date codominantly express two serum amyloid A (apoSAA) isoforms, apoSAA1 and apoSAA2, of which only apoSAA2 serves as a precursor for amyloid fibrils. In previous studies, we have shown that the CE/J strain, which produces a single, novel apoSAA isoform, apoSAACE/J, is amyloid resistant. In the present study amyloid-resistant CE/J females were mated with amyloid-susceptible CBA/J males to produce F1 hybrid offspring which were then backcrossed to the parental CBA/J mouse strain. Amyloid susceptibility was determined in 30 backcrossed mice 72 h after injection of murine amyloid enhancing factor and silver nitrate. ApoSAA isoforms in plasma were separated by isoelectric focusing gel electrophoresis and visualized after immunoblotting with anti-AA antiserum. Amyloid A fibrils in spleen homogenates were denatured by formic acid and AA protein was quantified by ELISA using anti-mouse apoSAA antibodies. Values < 5 apoSAA equivalent units were considered negative. 13 mice expressed an apoSAA1 and apoSAA2 doublet characteristic of CBA/J mice, whereas 17 mice, expressed the apoSAACE/J isoform codominantly with apoSAA1 and apoSAA2. The correlation of amyloid resistance to expression of the apoSAACE/J isoform was absolute (17/17 were negative; mean score 2.6 +/- 0.17 [standard error of the mean] apoSAA equivalent units) and the correlation between amyloid susceptibility and the expression of apoSAA2/apoSAA1 was also striking (12/13 were amyloid positive; mean score 47.9 +/- 9.0 [standard error of the mean] apoSAA equivalent units (P < 0.001). This is not significantly different from the 50% segregation of apoSAA phenotypes expected for linkage to a single gene. These results indicate that a single gene governs apoSAACE/J expression and thus confers protection against amyloid deposition even in the presence of apoSAA1 and apoSAA2 isoforms and show for the first time that resistance to AA amyloidosis is a dominant trait governed by a single gene.
Asunto(s)
Amiloidosis/genética , Genes Dominantes , Ligamiento Genético , Ratones Endogámicos/genética , Proteína Amiloide A Sérica/genética , Secuencia de Aminoácidos , Animales , Cruzamientos Genéticos , Femenino , Expresión Génica , Predisposición Genética a la Enfermedad , Masculino , Ratones , Ratones Endogámicos CBA/genética , Datos de Secuencia Molecular , Proteína Amiloide A Sérica/biosíntesis , Especificidad de la EspecieRESUMEN
During the acute phase response to tissue injury or inflammation, the concentration of several plasma proteins change. Previous work (29-34) suggested a role for interleukin 1 (IL-1) in the acute phase response. The availability of recombinant-generated mouse IL-1 prompted a study designed to directly test the function of IL-1 and its mechanism of action on hepatic synthesis of two positive acute phase proteins (serum amyloid A [SAA] and complement factor B), and a negative acute phase reactant (albumin). Intravenous injection of purified recombinant-generated murine-IL-1 into C3H/HeJ endotoxin-resistant mice induced a dose-dependent increase in SAA-specific hepatic messenger RNA (mRNA), and an increase in SAA plasma protein concentration. In primary murine hepatocyte cultures, both the recombinant IL-1 and highly purified human IL-1 induced a dose- and time-dependent, reversible increase in expression of the SAA and factor B genes, and a decrease in albumin gene expression. This regulation is pretranslational, since the kinetics and direction of change in specific mRNA for SAA, factor B, and albumin correspond to the changes in synthesis of the respective proteins. Moreover, the effect of IL-1 was specific, since actin gene expression was unaffected, and the IL-1 response was inhibited by antibody specific for IL-1. These data provide direct evidence that a single mediator, IL-1, can effect the positive and negative changes in specific hepatic gene expression characteristic of the acute phase response.
Asunto(s)
Regulación de la Expresión Génica/efectos de los fármacos , Inflamación/metabolismo , Interleucina-1/farmacología , Hígado/metabolismo , Biosíntesis de Proteínas , Proteínas de Fase Aguda , Animales , Proteínas Sanguíneas/biosíntesis , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Cinética , Masculino , Ratones , Ratones Endogámicos C3H , Proteína Amiloide A Sérica/biosíntesis , Especificidad de la EspecieRESUMEN
Secondary amyloidosis is a complication of diseases characterized by recurrent acute inflammation. In this study, a standardized stimulus which induced fever and inflammation was given to six normal subjects (19-24 yr old) to follow the fluctuation in concentration of serum amyloid A (SAA), the precursor of the secondary amyloid fibril protein. After a single intramuscular injection of etiocholanolone (0.3 mg/kg), blood samples were drawn twice a day for 12 days for determination of SAA by solid phase radioimmunoassay. From a base line of <100 mug/ml, the SAA concentration began rising within 12 h to a maximum value at about 48 h of 1,350-1,800 mug/ml in three males and 380-900 mug/ml in three females and returned to base line by 4-5 days. The SAA response showed a similar time response to C-reactive protein (CRP), a well-documented acute phase protein which was assayed semiquantitatively by capillary tube precipitin reaction. CRP, but not SAA, showed a quantitative correlation with the amount of fever induced by etiocholanolone. One subject exhibited a second rise in SAA and CRP concentrations after acute over-indulgence with alcohol, suggesting that acute liver damage may have caused an acute phase reaction. Thus, a controlled episode of fever and inflammation produced a prompt and prolonged elevation of SAA and CRP concentrations. Unlike SAA, CRP has not been implicated in the pathogenesis of amyloidosis, although its relationship to the P component of amyloid has recently been established.
Asunto(s)
Amiloide/sangre , Proteína C-Reactiva/metabolismo , Etiocolanolona/farmacología , Inflamación/sangre , Inflamación/inducido químicamente , Factores de TiempoRESUMEN
In plasma, the bulk of apoSAA, a positive acute phase reactant protein, is transported in high density lipoproteins (HDL), especially HDLH (apoA1-rich HDL). In this study we tested whether apoA1 deficiency would adversely affect apoSAA concentration and lipid distribution in mouse plasma lipoproteins. Acute phase response (APR) was induced in C57BL/6J (apoA1+/+) and apoA1-knockout mice (apoA1-/-) by a subcutaneous injection of silver nitrate. The APR increased cholesterol concentrations in LDL of apoA1-/- mice and apoA1+/+ mice in a like manner. In contrast to apoA1+/+ mice, concentrations of cholesterol, phospholipids and proteins in both HDLL (1.063Asunto(s)
Reacción de Fase Aguda
, Apolipoproteína A-I/deficiencia
, Apolipoproteínas/metabolismo
, Lipoproteínas HDL/sangre
, Proteína Amiloide A Sérica/metabolismo
, Animales
, Apolipoproteína A-I/genética
, Colesterol/sangre
, Cromatografía en Gel
, Electroforesis en Gel de Agar
, Lipoproteínas LDL/sangre
, Ratones
, Ratones Endogámicos C57BL
, Ratones Noqueados
, Fosfolípidos/sangre
, Triglicéridos/sangre
RESUMEN
CBA/J and other inbred strains of mice that express the amyloidogenic apolipoprotein serum amyloid A (apoSAA) apoSAA2, together with apoSAA1, are susceptible to amyloid A (AA) amyloidosis, whereas CE/J mice that express a single unique isoform, apoSAACEJ, are resistant. Studies indicate that CBA/JxCE/J hybrid mice that express apoSAA2 in the presence of apoSAACEJ are protected from amyloidogenesis. To define a mechanism by which expression of apoSAACEJ may protect from AA formation in the presence of apoSAA2, binding of recombinant apoSAA (r-apoSAA) isoforms, validated by N-terminal sequencing, to a murine macrophage cell line was investigated. Maximal specific binding occurred after incubation of radiolabeled apoSAA with IC-21 macrophages (1x105 cells/ml) for 30 min at 4 degreesC. The binding of 125I-r-apoSAA1, 125I-r-apoSAA2 and 125I-r-apoSAACEJ was specific and saturable, with an affinity (Kd) of about 2.8, 3.2 and 1.3 nM, respectively, and approximately 2-4x106 sites per cell. Competitive binding experiments indicate apoSAACEJ binds with higher affinity to macrophages than does either apoSAA1 or apoSAA2. We suggest that greater cellular affinity of apoSAACEJ compared to apoSAA2 may contribute to protection from AA amyloid in certain CBA/JxCE/J hybrid mice by interfering with interaction of apoSAA2 by macrophages and hence either membrane associated or intracellular degradation.
Asunto(s)
Amiloidosis/metabolismo , Apolipoproteínas/metabolismo , Macrófagos/metabolismo , Proteína Amiloide A Sérica/metabolismo , Animales , Susceptibilidad a Enfermedades , Ratones , Ratones Endogámicos CBA , Isoformas de Proteínas/metabolismoRESUMEN
BACKGROUND: Inflammation is an important component of ischemic heart disease. PTX3 is a long pentraxin whose expression is induced by cytokines in endothelial cells, mononuclear phagocytes, and myocardium. The possibility that PTX3 is altered in patients with acute myocardial infarction (AMI) has not yet been tested. METHODS AND RESULTS: Blood samples were collected from 37 patients admitted to the coronary care unit (CCU) with symptoms of AMI. PTX3 plasma concentrations, as measured by ELISA, higher than the mean+2 SD of age-matched controls (2.01 ng/mL) were found in 27 patients within the first 24 hours of CCU admission. PTX3 peaked at 7.5 hours after CCU admission, and mean peak concentration was 6.94+/-11.26 ng/mL. Plasma concentrations of PTX3 returned to normal in all but 3 patients at hospital discharge and were unrelated to AMI site or extent, Killip class at entry, hours from symptom onset, and thrombolysis. C-reactive protein peaked in plasma at 24 hours after CCU admission, much later than PTX3 (P<0.001). Patients >64 years old and women had significantly higher PTX3 concentrations at 24 hours (P<0.05). PTX3 was detected by immunohistochemistry in normal but not in necrotic myocytes. CONCLUSIONS: PTX3 is present in the intact myocardium, increases in the blood of patients with AMI, and disappears from damaged myocytes. We suggest that PTX3 is an early indicator of myocyte irreversible injury in ischemic cardiomyopathy.
Asunto(s)
Proteína C-Reactiva/metabolismo , Infarto del Miocardio/metabolismo , Componente Amiloide P Sérico/metabolismo , Anciano , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Infarto del Miocardio/sangre , Infarto del Miocardio/patología , Miocardio/metabolismo , Miocardio/patología , Necrosis , Concentración Osmolar , Valores de Referencia , Factores de TiempoRESUMEN
Activated astrocytes have been identified as the main source of the serine protease inhibitor alpha 1-antichymotrypsin (ACT), an acute phase protein that is tightly associated with amyloid plaques in Alzheimer's disease (AD) and in normal aged human and monkey brain. We analyzed the synthesis of ACT by cultured murine astrocytes in vitro. The murine astrocytes expressed an ACT-like antigen that crossreacted with antibodies to human ACT. The murine ACT-like protein is secreted by the astrocytes and is able to form an SDS-resistant complex with the serine protease cathepsin G, indicating that the secreted ACT is biologically active. We conclude that cultured primary astrocytes synthesize and secrete murine ACT in an active form. We, therefore, suggest that the ACT present within AD plaques is locally derived from plaque-associated activated astrocytes as a part of a glia-mediated local inflammatory response that is associated with the neurodegeneration seen in AD.
Asunto(s)
Astrocitos/metabolismo , alfa 1-Antiquimotripsina/biosíntesis , alfa 1-Antiquimotripsina/metabolismo , Animales , Animales Recién Nacidos/metabolismo , Especificidad de Anticuerpos , Química Encefálica/fisiología , Catepsina G , Catepsinas/metabolismo , Células Cultivadas , Humanos , Inmunohistoquímica , Ratones , Serina Endopeptidasas/metabolismo , alfa 1-Antiquimotripsina/químicaRESUMEN
A solid-phase, direct binding ELISA for serum amyloid A (SAA) proteins is described, in which noncovalent interactions of SAA with other plasma constituents are disrupted to permit direct coating of the wells of flexible polyvinyl chloride microtitration plates with an amount of SAA antigen proportional to its concentration in plasma. The wells are coated overnight at 60 degrees C with plasma diluted in 3 M potassium bromide and 0.1 M sodium bicarbonate. pH 9.6. The next day, any remaining sites on the wells are blocked by incubation for 1 h at ambient temperature with a 5% solution of dry milk solids and 0.05% Tween 20 in 0.02 M phosphate buffer, pH 7.4. The wells are rinsed and incubated for 90 min at 37 degrees C with polyclonal rabbit or rat anti-human SAA antiserum. Then, the wells are rinsed and incubated with goat anti-rabbit or rat IgG antiserum to which has been conjugated horseradish peroxidase. o-phenylenediamine and hydrogen peroxide substrates are added to the wells, color is allowed to develop, and sulfuric acid is added to stop the enzyme-catalyzed reaction. The amount of SAA coated to wells is quantified by absorbance at 490 nm. Four or more serial three-fold dilutions of plasma samples are assayed simultaneously on separate plates. Each plate contains a set of wells with identical concentrations of SAA standard protein diluted in decreasing concentrations of plasma proteins corresponding to the dilution of sample. The method can detect SAA concentrations in plasma samples ranging from 1 microgram/ml to greater than 1000 micrograms/ml. The method is suited to serial monitoring of SAA concentration in patients undergoing treatment for inflammatory conditions and to the quantitative analysis of large numbers of samples.
Asunto(s)
Proteína Amiloide A Sérica/análisis , Western Blotting , Relación Dosis-Respuesta Inmunológica , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/normas , Humanos , Peso Molecular , Albúmina Sérica/metabolismo , TemperaturaRESUMEN
Murine serum amyloid P component (SAP) is an acute-phase protein that is increased 2-10-fold in concentration following appropriate inflammatory or infectious stimuli. Previous studies of the acute-phase SAP response have employed quantitative immunoelectrophoresis or radioimmunoassay to measure SAP concentration. A rate nephelometric procedure has been developed which measures SAP concentration rapidly and with equivalent or greater precision than the previously applied techniques. This simple method will facilitate experimental and clinical studies of the acute-phase response.
Asunto(s)
Amiloide/sangre , Nefelometría y Turbidimetría , Proteínas de Fase Aguda , Amiloide/inmunología , Animales , Reacciones Antígeno-Anticuerpo , Proteínas Sanguíneas/análisis , Femenino , Inmunoelectroforesis , Ratones , Ratones Endogámicos CBA , Precipitinas/análisis , Conejos , Radioinmunoensayo , Componente Amiloide P SéricoRESUMEN
The joint destruction of osteoarthritis (OA) comprises loss of articular cartilage resulting from an imbalance of enzyme-catalized cartilage breakdown and regeneration. OA is thought to derive from defective chondrocyte metabolism and thus to inherently lack the large-scale systemic response that is the hallmark of rheumatoid arthritis (RA). Because of the apparent absence of systemic inflammation in OA, acute-phase response proteins have not been as extensively studied in OA as they have been in RA. The diagnosis of OA almost always involves radiographic assessment of joint damage, which is useful only after the disease process has been underway for several months. Radiographic evaluation cannot give a good assessment of current disease activity and is a relatively insensitive indicator of prognosis. Cartilage breakdown products can potentially serve as direct surrogate markers of OA disease activity, but have not been extensively used because of their limited sensitivity and the technical difficulties associated with their measurement. Markers of disease activity in RA are indirect and are derived from the acute-phase response, a cycle of temporal changes in cellular and metabolic function. The early part of the acute-phase response involves the local action and production of cytokines such as interleukin-1 (IL-1), tumor necrosis factor (TNF-alpha) and IL-6. In the late acute-phase response, these cytokines can effect many systemic changes, including increased production of acute-phase proteins (APP). Three valuable surrogate markers of disease activity in RA are provided by the acute-phase response: the time-honored erythrocyte sedimentation rate (ESR) and the newer APPs C-reactive protein (CRP) and serum amyloid A (SAA). As in RA, the joint destruction of OA involves IL-1, TNF-alpha, and IL-6; however, OA can be viewed as an indolent stimulus of the later (systemic) acute-phase response. Recent studies of the acute-phase response in OA suggest that the concentrations of CRP and SAA are elevated in OA, but to a lesser extent than in RA. In the future, long-term monitoring of CRP concentrations in the blood may permit the earlier detection and more effective treatment of OA.
Asunto(s)
Proteínas de Fase Aguda/análisis , Osteoartritis/metabolismo , Biomarcadores/análisis , Sedimentación Sanguínea , Proteína C-Reactiva/análisis , Citocinas/análisis , Citocinas/biosíntesis , Humanos , Osteoartritis/sangre , Osteoartritis/diagnóstico , Osteoartritis/tratamiento farmacológico , Proteína Amiloide A Sérica/análisis , Líquido Sinovial/química , Factores de TiempoRESUMEN
Serum amyloid A (SAA) is a major inducible acute phase protein characterized as a transient injury specific constituent of high density lipoprotein. We investigated whether the acute phase SAA (A-apoSAA), as a marker of inflammation, is present in the brain of monkeys with surgically induced hypertension of 39 months duration. Sections from brains of normotensive monkeys (systolic blood pressure < 124 mmHg) and hypertensive monkeys (systolic blood pressure > 185 mmHg) were processed for immunohistochemistry with a rabbit polyclonal antiserum to human A-apoSAA. We found that A-apoSAA was present in hypertensive but not in normotensive brain sections. Staining was localized to capillary endothelial cells and occasionally to the entire vessel wall of the prefrontal cortex. Staining was also observed in the capillaries and in medium size vessels of the corona radiata, the head of the caudate and, to a smaller extent, in the putamen. Additionally, the A-apoSAA was present in cells forming a circular configuration within microinfarcts. These findings suggest that high blood pressure in the brain can result in either local production of A-apoSAA in the capillaries and within microinfarcts or uptake of A-apoSAA from the blood
Asunto(s)
Apolipoproteínas/metabolismo , Infarto Encefálico/metabolismo , Capilares/metabolismo , Hipertensión/metabolismo , Proteína Amiloide A Sérica/metabolismo , Animales , Encéfalo/irrigación sanguínea , Encéfalo/metabolismo , Encéfalo/patología , Infarto Encefálico/patología , Capilares/patología , Humanos , Hipertensión/patología , Inmunohistoquímica , Macaca mulatta , Precursores de Proteínas/metabolismo , ConejosRESUMEN
OBJECTIVE: To evaluate ferritin concentration in serum and synovial fluid (SF) as a marker of activity of arthritis in comparison with C-reactive protein (CRP) and acute-phase serum amyloid A protein (A-SAA). METHODS: We determined the concentrations of ferritin, CRP and A-SAA in paired serum and SF in 34 rheumatoid arthritis (RA) and 21 osteoarthritis (OA) patients. The erythrocyte sedimentation rate (ESR) was also measured. RESULTS: The serum concentrations of ferritin, CRP and A-SAA were 93 +/- 76 (mean +/- SD) ng/ml, 4 +/- 5 mg/ml, 8 +/- 4 mg/ml in OA and 140 +/- 227, 59 +/- 34, 289 +/- 223 in RA, respectively. There was no significant difference in serum ferritin levels between OA and RA, and serum ferritin did not correlate with ESR, CRP or A-SAA. Both serum CRP and A-SAA levels were significantly higher in RA than in OA (p < 0.0001, p < 0.0001), and correlated with ESR in all arthritis (r = 0.658, p < 0.0001, r = 0.404, p < 0.01), respectively. Serum CRP levels correlated with A-SAA levels in serum (r = 0.727, p < 0.0001). In SF, the concentrations of ferritin, CRP and A-SAA in RA (421 +/- 307, 25 +/- 20 and 39 +/- 41) were significantly higher (p < 0.01, p < 0.0001, p < 0.001) than those in OA (202 +/- 220, 2 +/- 2 and 2 +/- 2), respectively. There were significant correlations among SF ferritin, CRP and A-SAA. CONCLUSION: Ferritin levels in SF but not in serum are significantly elevated in RA more than in OA, and ferritin correlated with CRP or A-SAA in SF, but not in serum. Higher levels of SF ferritin, as well as SF CRP and SF A-SAA, seem to reflect greater degrees of joints inflammation in RA and OA.
Asunto(s)
Apolipoproteínas/análisis , Proteína C-Reactiva/análisis , Ferritinas/análisis , Proteína Amiloide A Sérica/análisis , Líquido Sinovial/química , Reacción de Fase Aguda/sangre , Reacción de Fase Aguda/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Artritis Reumatoide/sangre , Artritis Reumatoide/metabolismo , Biomarcadores , Sedimentación Sanguínea , Femenino , Ferritinas/sangre , Humanos , Masculino , Persona de Mediana Edad , Osteoartritis/sangre , Osteoartritis/metabolismoRESUMEN
Immunohistochemical localization of the injury specific apolipoprotein, acute phase serum amyloid A (A-apoSAA), was compared in brains of patients with neuropathologically confirmed Alzheimer's disease (AD), multiple sclerosis (MS), Parkinson's disease (PD); Pick's disease (Pick's), dementia with Lewy bodies (DLB), coronary artery disease (CAD), and schizophrenia. Affected regions of both AD and MS brains showed intense staining for A-apoSAA in comparison to an unaffected region and non-AD/MS brains. The major site of A-apoSAA staining in both diseases was the myelin sheaths of axons in layers V and VI of affected cortex. A-apoSAA contains a cholesterol binding site near its amino terminus and is likely to have a high affinity for cholesterol-rich myelin. These findings, along with our recent evidence that A-apoSAA can inhibit lipid synthesis in vascular smooth muscle cells suggest that A-apoSAA plays a role in the neuronal loss and white matter damage occurring in AD and MS.
Asunto(s)
Enfermedad de Alzheimer/metabolismo , Apolipoproteínas/metabolismo , Encéfalo/metabolismo , Vaina de Mielina/metabolismo , Terminales Presinápticos/metabolismo , Proteína Amiloide A Sérica/metabolismo , Anciano , Anciano de 80 o más Años , Enfermedad de Alzheimer/patología , Encéfalo/patología , Membrana Celular/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Precursores de Proteínas/metabolismoRESUMEN
The acute phase SAA and SAP profiles have been compared for localized and endotoxin induced inflammation in LPS responder and nonresponder strains of mice. The SAP profile can reflect a delay with respect to the start of the increase. Its maximum is on the order of ten times the nonacute phase concentration and elevated concentrations are sustained 24 to 48 hours after SAA concentration is rapidly decreasing to normal. The role of Interleukin 1, known to have an essential role in SAA production, was investigated for SAP production. Purified mouse IL 1 and rabbit IL 1 produced a minimal elevation of SAP concentration above normal values, especially when compared with their effects on SAA concentration. BCG infection was shown to synergistically augment SAA induction by LPS and was shown to enhance IL 1 production by macrophages in response to LPS. Unlike SAA synthesis, BCG-preinfection fails to synergistically augment the LPS-induced SAP response. BCG infection alone produced highly elevated and sustained increases in SAP concentration, whereas, the effect on SAA concentration was minimal. Macrophages appear to play an important role in SAP acute phase elevation, but the mechanism in different from that of SAA elevation.
Asunto(s)
Amiloide/biosíntesis , Inflamación/metabolismo , Proteínas/metabolismo , Proteína Amiloide A Sérica/biosíntesis , Enfermedad Aguda , Animales , Caseínas/farmacología , Endotoxinas/farmacología , Escherichia coli , Femenino , Interleucina-1 , Ratones , Ratones Endogámicos C3H , Mycobacterium bovis , Radioinmunoensayo , Componente Amiloide P Sérico , Nitrato de Plata/farmacología , Tuberculosis/metabolismoRESUMEN
Acute phase serum amyloid A (A-apoSAA), but not constitutive apoSAA (C-apoSAA), was identified by Western blotting experiments in brain protein extracts from eight of nine patients with Alzheimer's disease (AD), one with a brain tumor and one with multiple sclerosis. A-apoSAA was not detected in six subjects with Pick's or Lewy Body disease or three other non-AD brain specimens. Apolipoprotein A-I and albumin were not found in any of the brain protein extracts. A-apoSAA mRNA was detected in AD brain by reverse transcription-polymerase chain reaction (RT-PCR). These data suggest that apoSAA is locally produced in AD brain and that investigation of the neuroinflammatory effects of this injury specific apolipoprotein is warranted.
Asunto(s)
Enfermedad de Alzheimer/metabolismo , Apolipoproteínas/biosíntesis , Apolipoproteínas/metabolismo , Encéfalo/metabolismo , Proteína Amiloide A Sérica/metabolismo , Anciano , Western Blotting , Femenino , Humanos , Masculino , Factores de TiempoRESUMEN
In 106 patients with systemic amyloidosis (56 primary, 27 secondary, and 23 familial), serum amyloid A protein (SAA) was measured by solid-phase radioimmunoassay and C-reactive protein (CRP) was measured by rate nephelometry. SAA and CRP concentrations were highly correlated (r = 0.75, P less than 0.001) throughout the normal and abnormal concentration ranges. In systemic amyloidosis, SAA was more sensitive than CRP as an indicator of the acute-phase response, particularly in secondary amyloidosis. Acute-phase proteins are only occasionally increased during the course of familial amyloidosis. The overlap of acute-phase protein levels does not permit reliable separation of primary amyloidosis from secondary amyloidosis solely on the basis of such studies despite the significantly higher SAA and CRP levels in the latter.
Asunto(s)
Amiloide/sangre , Amiloidosis/sangre , Proteína C-Reactiva/sangre , Proteína Amiloide A Sérica/sangre , Amiloidosis/genética , Humanos , RadioinmunoensayoRESUMEN
Against a background of growing interest in more sensitive assays for quantifying various acute phase proteins, we evaluated the performance of recently developed tests for C-reactive protein (CRP), serum amyloid A (SAA) and mannose-binding protein (MBP) on the Behring nephelometer II (BNII). Sample results outside the calibration ranges of 3.5 to 220 mg/L for CRP, 3.3 to 215 mg/L for SAA and 0.09 to 5.6 mg/L for MBP were automatically re-measured at another dilution. The lower limits of detection were 0.01, 0.7 and 0.01 mg/L for CRP, SAA and MBP, respectively. The coefficients of variation (CV) for intra- (n > or = 20) and inter- (n > or = 15) assay precision were < 5.2% and < 8.5%, respectively, for the three proteins at concentrations representing low, normal and high. Linearity for each method was within 5% of the expected values throughout the calibration range. We observed no significant interference from bilirubin (up to 300 mg/L) or haemoglobin (up to 10 g/ L) for the three tests. Method comparison studies performed for CRP and SAA yielded the following results: y (CRP on BNII) = 0.75x (ELISA, Hemagen) -0.25 mg/L (r = 0.981, Sy/x = 2.1 mg/L; y (SAA on BNII) = 1.44x (ELISA, Hemagen) -9.9 mg/L (r = 0.972, Sy/x = 6.9 mg/L), where ELISA is enzyme-linked immunosorbent assay. Reference intervals established in 261 adult blood donors (aged 36.2 +/- 9.0 years) were found to be log-normal with 2.5th, 50th, and 97.5th centiles of < 0.17, 1.00 and 10.1 mg/L for CRP, < 0.84, 2.10 and 9.70 mg/L for SAA; and 0.30, 1.28 and 4.10 mg/L for MBP. We observed no relationship with CRP concentration and age; however, SAA levels increased with age while MBP levels decreased. The BNII provides a simple, rapid and sensitive system for measuring CRP, SAA and MBP in human serum.
Asunto(s)
Apolipoproteínas/análisis , Proteína C-Reactiva/análisis , Proteínas Portadoras/sangre , Nefelometría y Turbidimetría/métodos , Proteína Amiloide A Sérica/análisis , Adulto , Ensayo de Inmunoadsorción Enzimática , Estudios de Evaluación como Asunto , Humanos , Sueros Inmunes , Lectinas de Unión a Manosa , Estándares de Referencia , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Hepatic levels of mRNA specific for total serum amyloid A (SAA), the SAA1 and SAA2 isotypes, serum amyloid P (SAP), C-reactive protein (CRP), and fibronectin, as well as the plasma concentrations of SAA and SAP were examined in amyloid-resistant (A/J) and amyloid-susceptible (CBA/J) mice during azocasein-induced chronic inflammation. In both strains hepatic SAA and SAP mRNA levels and plasma SAA and SAP protein concentrations increased dramatically during the early stages of inflammation; this was followed by a decrease to concentrations that were maintained at levels considerably higher than background. The ratios of SAA1 and SAA2 mRNA and plasma protein were 1:1 throughout. This indicated that there was no preferential accumulation of mRNA specifying a particular isotype and no preferential synthesis or clearance of a particular isotype during chronic inflammation and the early stages of amyloidogenesis in either strain. Similarly, hepatic SAP mRNA levels in both strains increased dramatically during the early stages of inflammation and were subsequently maintained at elevated levels. Plasma SAP concentrations increased rapidly during the first three days of the study in both A/J and CBA/J mice; however, during the later stages of inflammation, A/J plasma SAP levels decreased to a steady-state concentration that was approximately half that observed in CBA/J mice. Our results identify differences in the hepatic mRNA and plasma protein levels of the major mouse acute-phase reactants (APR) in the amyloid-resistant A/J and amyloid-susceptible CBA/J mouse strains. These findings are consistent with circulating inflammatory APR concentrations contributing, together with other factors, to the onset and pathogenesis of secondary amyloidosis.