,
* "Antonio Taticchi" Unit for Atherosclerosis and Thrombosis, Istituto Ricerche Farmacologiche Mario Negri, Consorzio Mario Negri Sud, Italy;
Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy;
Istituto di Patologia, Università di Milano, Italy; and
Centro di Ricerche e Alta Formazione, Università Cattolica, Campobasso, Italy
1Correspondence: "Antonio Taticchi" Unit for Atherosclerosis and Thrombosis, Istituto Ricerche Farmacologiche Mario Negri, Consorzio Mario Negri Sud, Via Nazionale, 66030 S. Maria Imbaro, Italy. E-mail: lorenzet{at}negrisud.it
 TOP ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
, PTX3 was not effective. The increase in TF activity requires mRNA synthesis, as assessed by polymerase chain reaction. The mechanism by which PTX3 modulates TF synthesis resides in an enhanced I
B, phosphorylation and degradation and increased migration of the transacting factor c-Rel/p65 into the nucleus, as determined by Western blot and electro-mobility shift assay. These results show that PTX3 is an enhancer of the expression of TF by mononuclear cells. In the area of vascular injury, during the inflammatory response, cell-mediated fibrin deposition takes place. PTX3 increases TF expression, thus potentially playing a role in thrombogenesis and wound healing.
Key Words: thrombogenesis • leukocytes • acute phase • fibrin formation
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
More recently, larger proteins, in which the C-terminal domain is highly homologous with the sequence of the classical PTXs, have been identified. In these molecules, the C-terminal half is coupled with a long, unrelated N-terminal portion, which increases their size of approximately twice as compared with the "short" PTXs. These proteins were named "long" PTXs. PTX3, the first member of this subfamily to be discovered, was cloned as an interleukin-1 (IL-1)- and tumor necrosis factor (TNF)-inducible gene in endothelial cells and fibroblasts [2 ].
Although SAP and CRP are made in the liver in response to inflammatory mediators, most prominently IL-6 [1 ], PTX3, is predominantly expressed at extrahepatic sites and is found elevated in muscular tissues, including the heart [3 ].
PTX3 binds the classical complement component C1q [4 ] and selected microorganisms [5 ], suggesting a role for PTX3 in the amplification of inflammation and in the innate immune response.
PTX3 production is induced in models of systemic or localized infection including sepsis [6 ]. In addition, dramatic elevations of PTX3 were observed in patients with sepsis [7 ]. Very recently, PTX3 was found increased in the blood of patients with acute myocardial infarction [8 ]. The finding that a strong expression of PTX3 could be detected in advanced atherosclerotic lesions suggests that PTX3 may also contribute to the pathogenesis of atherosclerosis [9 ].
Nonvascular cells constitutively express tissue factor (TF), the in vivo trigger of blood coagulation. Upon vascular injury, TF is exposed to circulating blood, where it becomes available to the binding of factor VIIa. The resulting complex, activating factors IX and X, leads to cleavage of prothrombin and ultimately, fibrin formation. A growing body of evidence assigns TF a central role in thrombosis and inflammation associated with cancer, sepsis, and atherosclerosis [10 ].
Although normally absent in intravascular cells, TF can be induced in circulating monocytes and endothelial cells by a wide variety of agents, including lipopolysaccharide (LPS) [11
, 12
], immune complexes [13
], inflammatory cytokines [14
], and P-selectin [15
]. A mechanism that involves binding of c-Rel/p65 heterodimers to a putative B site in the TF promoter is responsible for its synthesis [16
].
We recently showed that PTX3 functions as a modulator of TF synthesis in activated endothelial cells [17 ], and the effect of PTX3 on TF expression by human monocytes remains unknown. In the present study, we demonstrate that PTX3 enhances the expression of TF on LPS-stimulated mononuclear cells.
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
were from PeproTech (Rocky Hill, NJ). SAP and CRP (
94% pure) were from Sigma Chemical Co. (St. Louis, MO). The monoclonal anti-TF antibody HTF1 was a generous gift of Dr. Yale Nemerson (Mt. Sinai School of Medicine, New York). Sterile, pyrogen-free microtubes were obtained from Sarstedt (Numbrecht, Germany).
Cell isolation and culture
Human mononuclear cells (MN) were obtained from whole blood collected from healthy donors by Lymphoprep sedimentation as described previously [18
]. The monocytes in this population were 25–30%, as assessed by nonspecific esterase staining. Monocytes were purified by using a discontinuous Percoll density gradient [18
]. The Percoll-isolated fraction contained 85% monocytes. Cells were resuspended in RPMI-1640 medium supplemented with penicillin (100 U/mL) and streptomycin (100 µg/mL) at a cell concentration of 3 x 106/mL in the presence of LPS, rhIL-1ß, rhTNF-, SAP, CRP, or human PTX3, purified from the supernatant of transfected Chinese hamster ovary cells as described [4
], alone or in different combinations in sterile, pyrogen-free, stoppered test tubes at 37°C.
Control of LPS contamination
Sterile, pyrogen-free working conditions were observed to avoid any contamination by LPS. Solutions were prepared in glassware rendered pyrogen-free by heating at 180°C for 3 h. Reagents were dissolved in sterile pyrogen-free solvents and when tested for LPS contamination by the Limulus assay (Whittaker Bioproducts, Inc., Walkersville, MD), were found negative at a sensitivity threshold of 0.1 LPS unit/ml, corresponding to 0.01 ng/mL.
TF assays
After incubation, cells were disrupted by three freeze-thaw cycles, and procoagulant activity was assessed by a one-stage clotting time [18
]. Results were expressed in arbitrary units (U) by comparison with a standard curve obtained using a human brain thromboplastin standard kindly donated by Dr. Leon. Poller (University of Manchester, Manchester, UK). This preparation was assigned a value of 1000 U for a clotting time of 20 s.
To determine TF antigen, cells were harvested and lysed in Tris-buffered saline (TBS), pH 8.5, containing 1% Triton X-100 overnight. Cell debris was pelleted by centrifugation at 100,000 g for 60 min at 4°C. The protein content of the supernatant was determined by a bicinchoninic acid protein assay kit (Pierce, Rockford, IL) and adjusted to 2 mg/ml. Samples were then used for enzyme-linked immunosorbent assay (ELISA; Imubind TF kit, American Diagnostica, Stamford, CT), according to the manufacturer’s instructions.
Polymerase chain reaction (PCR) analysis of TF mRNA
Oligonucleotides F1 (sense bp 178–198) and R1 (antisense bp 495–515), from the coding sequence of the human TF, and GF1 (sense bp 64–86) and GR1 (antisense bp 581–603), from the coding sequence of the human glyceraldehyde phosphate dehydrogenase (GAPDH), were synthesized.
To obtain the first cDNA strand, 1 µg total RNA from mononuclear leukocytes was reverse-transcribed using random hexamers and Moloney murine leukaemia virus reverse transcriptase (RT) [15 ]. PCR was performed with 5 µl cDNA in 10 mM Tris-HCl (pH 8.3), 50 mM KCl, 2 mM MgCl2, 0.4 µg each appropriate sense and antisense primers, 200 µM each dNTPs, and 2.5 units Taq polymerase. The amplification conditions were 94°C for 1 min, 60° for 1 min, and 72°C for 1 min to obtain a product of 528 bp from the GAPDH mRNA and 337 bp from the TF mRNA. The PCR was performed in the exponential phase (cycles 23 for GADPH or 32 for TF), as assessed in previous experiments, in which at various cycles, the PCR product was tested (data not shown). The reactions (8 µl) were analyzed on 1% agarose gel stained with ethidium bromide.
Electro-mobility shift assay (EMSA)
To determine the effect of PTX3 on c-Rel/p65 nuclear translocation, nuclear extracts from 4–5 x 106 MN cultured for 1 h with the different reagents were prepared, and the level of c-Rel/p65 was monitored by EMSA as described previously [19
]. Generally, protein concentrations, as determined by Bradford, ranged between 25 and 50 µg/4–5 x 106 MN. The TF-B-like probe (5'-GTC CCG GAG TTT CCT ACC GGG-3'; Labtek S.R.L., Corsico, Italy) was annealed with a complementary primer and radiolabeled with [32P]deoxycytidine 5'-triphosphate. Nuclear extracts (5 µg) were preincubated with 18 µmol/l binding buffer (60 mmol/l HEPES, pH 7.9, 180 mmol/l KCl, 3 mmol/l EDTA, 36% glycerol, 4 µg poly(dI-dC), 15 mmol/l dithiothreitol) in ice for 20 min. After preicubation, radiolabeled DNA probes (1x105 cpm) were added, and the mixture was incubated at room temperature for 20 min. Examination of DNA binding proteins was performed on electrophoresis on native 5% acrylamide gels in 0.5x Tris-boric acid-EDTA buffer. Autoradiography was performed on Kodak XAR films for 16–48 h at –80°C.
Western blot analysis
Following experimental treatment, cytoplasmic extracts were prepared. The extracts (100 µg total proteins) were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto nitrocellulose membrane in 25 mM Tris/192 mM glycine/20% methanol for 3 h at 80 V and 4°C as described previously [20
]. The blots were blocked for 1 h in 20 mM Tris, pH 7.6/137 mM NaCl/0.1% Tween-20 [TBS/Tween 20 (TBST)] containing 5% dry milk. The blots were washed in TBST and incubated with primary antibody for inhibitor of IB (IB; Santa Cruz Biotechnology, Santa Cruz, CA) for 1 h at room temperature with shaking. After washing with TBST, the blots were incubated with a secondary antibody, and proteins were visualized by chemiluminescence (Amersham Pharmacia Biotech). Ponceau S staining of membranes monitored transfer efficiency.
Statistical analysis
The results are given as mean values ± SEM. Differences between groups were tested for significance using Student’s t-test for paired observations unless otherwise indicated. ANOVA analysis followed by Dunnett’s test was used for multiple comparisons.
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
When purified PTX3 was present during the incubation, TF activity was enhanced in a concentration-dependent way, reaching approximately a threefold increase at a concentration of 10 µg/mL (Fig. 1A ). No effect could be observed when PTX3 was incubated with MN alone in the absence of other stimuli (Fig. 1A) .
 View larger version (10K): [in a new window] |
Figure 1. Effect of PTX3 on TF expression in mononuclear cells and monocytes. (A) TF expression was measured in MN, which were exposed to different concentrations of PTX3 without ( ) or with ( ) LPS (0.1 µg/mL) for 6 h at 37°C (n=4, mean±SEM; *, P<0.01; **, P<0.002; ***, P<0.001). (B) TF activity from monocytes (M) alone (C) or exposed to LPS (0.1 µg/mL) without or with PTX3 (5 µg/mL) for 6 h at 37°C (n=3, mean±SEM;*, P<0.05, for LPS+PTX3 vs. LPS). At the end of incubation, samples were frozen and thawed, and TF activity was measured by a one-stage clotting time, as described in Materials and Methods.
|
Although among mononuclear leukocytes, the monocyte is the only cell known as capable of expressing TF, we confirmed that the cellular target for PTX3 up-regulation of TF was the monocyte. Monocytes isolated from MN with a Percoll density gradient incubated with LPS expressed TF activity that was enhanced in the presence of PTX3 (Fig. 1B) .
To test whether PTX3 was effective also when agents different from LPS were used to stimulate monocytes, other known inducers of TF activity, namely IL-1ß and TNF-, were incubated with the cells. Maximal effective concentrations of the inducers were determined by dose responses (not shown) and used in these experiments. The almost no-detectable activity expressed by MN alone (0.03±0.01 U/3x105 MN) was greatly enhanced by the different agents (Fig. 2
). PTX3 was effective only when LPS was used. No increment in activity was detected when LPS was replaced by IL-1ß or TNF- (Fig. 2)
.
 View larger version (7K): [in a new window] |
Figure 2. Effect of PTX3 on TF expression in MN exposed to LPS, IL-1ß, and TNF-. MN were incubated with LPS (0.1 µg/mL), IL-1ß (40 ng/mL), or TNF- (40 ng/mL) at the presence of PTX3 (5 µg/mL). Treatment conditions were as outlined in the legend of Figure 1
. The data represent the mean ± SEM of three duplicate experiments. *, P < 0.03. TFa, TF activity.
|
 View larger version (13K): [in a new window] |
Figure 3. Effect of PTX3, CRP, and SAP on TF expression in MN, which were exposed to SAP, CRP, or PTX3 (5 µg/mL each) with (open bars) or without (solid bars) LPS (0.1 µg/mL). Conditions are the same as in Figure 1
. Bars represent the mean of four experiments ± SEM. *, P < 0.02.
|
Regulation of TF mRNA levels in LPS-stimulated MN by PTX3
To examine the effect of PTX3 on TF mRNA, MN were incubated with or without LPS in the presence and in the absence of PTX3 for 1 h, after which the cells were washed and treated to isolate mRNA. The mRNA was then reverse-transcribed and used for parallel assay of TF and GAPDH mRNA by PCR amplification. The expected 337-bp PCR product for TF was obtained. Southern blot analysis of the PCR products was performed. No PCR product from the control cells could be detected (Fig. 4
). In contrast, expression of TF mRNA could be observed in cells exposed to LPS. PTX3, which by itself had no effect on MN (not shown), caused a strong increase in the level of expression of TF mRNA. Southern blot analysis of GAPDH mRNA showed similar mRNA levels in control and LPS- and PTX3-treated cells, indicating that the efficiency of RT was comparable among the experimental groups.
 View larger version (19K): [in a new window] |
Figure 4. Effect of PTX3 on TF mRNA levels in LPS-stimulated MN, which untreated (C) or exposed to LPS (0.1 µg/mL) with or without PTX3 (5 µg/mL) for 1 h at 37°C, were treated for RT-PCR, and analysis of TF and GAPDH mRNA expression was assessed by ethidium bromide staining. Data are representative of three separate experiments.
|
 View larger version (60K): [in a new window] |
Figure 5. Effect of PTX3 on nuclear translocation of c-Rel/p65 in LPS-stimulated MN. Nuclear extracts were prepared from MN untreated (C) or treated with LPS (0.1 µg/mL), with or without PTX3 (5 µg/mL), for 30 min at 37°C. The presence of c-Rel/p65 heterodimers was determined by EMSA. Data are representative of three separate experiments.
|
BB. We investigated the effect of PTX3 on degradation of IB in MN stimulated with LPS. Cytoplasmic proteins were extracted from treated MN and analyzed by Western blot. As expected, LPS treatment resulted in rapid degradation of IB (Fig. 6
). PTX3 greatly increased the proteolytic degradation of IB in stimulated MN.
 View larger version (20K): [in a new window] |
Figure 6. Effect of PTX3 on IB. MN untreated (C) or incubated with LPS (0.1 µg/ml) for 5 and 15 min with or without PTX3 (5 µg/mL) were treated for cytosolic protein extraction. The samples were subjected to analysis by Western blotting using an IkB-specific antibody. Data are representative of three separate experiments.
|
B degradation in LPS-stimulated Percoll-purified monocytes, with an effect similar to that exerted in MN (not shown). |
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
PTX3, a member of the family of the PTXs, is expressed in several cell types, most prominently MN and endothelial cells, following stimulation by LPS, IL-1ß, and TNF- [2
]. PTX3 production is induced in models of systemic or localized infection including sepsis [6
]. Recently, plasma levels of PTX3 were found high in critically ill patients where they correlated with the severity of the disease (with a gradient from systemic inflammatory response syndrome to septic shock), the highest levels of PTX3 being associated with an unfavorable outcome [7
].
Under physiological conditions, TF is exclusively expressed by extravascular cells. However, expression of TF has been detected on monocytes and endothelial cells in vitro and in vivo in response to stimulation, e.g., by bacterial LPS; indeed, TF expression by monocytes is one of the leading causes of disseminated intravascular coagulation in patients with severe sepsis [23 ]. When exposed to inflammatory agents, monocytes and endothelial cells express TF on their membrane, shifting the hemostatic balance of the system toward a procoagulant state, which may lead to fibrin deposition, a common feature of inflammatory diseases.
Clinical evidences for the role of monocyte/macrophage TF in human disease comes from patients with meningococcal sepsis [23 ], peritonitis [24 ], lupus erythematosus [25 ], cirrhosis [26 ], Crohn’s disease [27 ], obstructive jaundice [28 ], adult respiratory distress syndrome [29 ], and active, unstable angina [30 ].
The elevated levels of PTX3 in the early stage of the inflammatory disease support the hypothesis of a role of PTX3 in the modulation of monocyte procoagulant activity.
PTXs are markers of the acute-phase response. SAP and CRP, which belong to the same family, are acute-phase reactants found elevated in plasma during unstable coronary syndromes [31 ]. Recently, PTX3 was found increased in the blood of patients with acute myocardial infarction [8 ]. PTX3 was detected in the intact myocardium and disappeared from damaged myocytes, suggesting that dying or necrotic cardiomyocytes could be the source of PTX3. In these patients, PTX3 peaked at 7.5 h, much earlier than the short PTX CRP. The lack of correlation with the levels of CRP suggests that PTX3 may represent a new and independent indicator of inflammatory components in ischemic heart disease.
Although the concentration of PTX3 used in our experiments far exceeds the amount of PTX3 in blood of patients with acute myocardial infarction, it is conceivable to speculate that during the onset of inflammatory reactions, recruitment of PTX3-producing cells such as monocytes will be responsible for delivering high amounts of this acute-phase reactant in a localized environment.
The effect of PTX3 was specific, as CRP and SAP were inactive in our test system. It is interesting that we could not detect TF activity when CRP was incubated with MN in the absence of LPS, unless used at exceedingly high concentrations, i.e., over 50 µg/mL. This result is in disagreement with previous reports, which showed induction of monocyte TF by CRP in the absence of other exogenously added stimuli with much lower concentrations [32 , 33 ]. Conversely, lately, Paffen et al. [34 ] have observed that human monocytes were almost insensitive to CRP in terms of TF expression and responded with a sixfold increase in monocyte chemoattractant protein-1 levels. A different degree of purity of CRP preparations and different experimental conditions could help us to understand this discrepancy. Potential LPS contamination in our experimental conditions has been avoided through very stringent laboratory techniques and confirmed by a commercial ELISA kit for LPS detection.
The effect of PTX3 is exerted at the level of mRNA as well as at the level of protein synthesis. Following LPS stimulation, monocytes initiate transcription of TF, which implies binding of the transcription factors activated protein-1 and the heterodimer c-Rel/p65, components of the nuclear factor (NF)-B transacting factor family, to the promoter of TF. In unstimulated monocytes, c-Rel/p65 is bound to IB, which holds the c-Rel/p65 transacting factor in an inactive state in the cytoplasm. Upon binding of LPS to its receptors on the cell membrane, protein kinase cascades are activated, phosphorylation of IB occurs, and the transacting factor c-Rel/p65 rapidly migrates to the nucleus [35
]. The results obtained in the present study are in agreement with the above picture: Incubation of LPS with MN induces degradation of IB and the consequent migration of c-Rel/p65. The presence of PTX3, together with LPS throughout the incubation, is responsible for an increased migration of c-Rel/p65 into the nucleus, which occurs by means of an enhanced degradation of IB.
Binding sites for the components of the NF-B/Rel superfamily are found in the promoter region of numerous proinflammatory genes and could play a key role, among others, in the pathogenesis of inflammatory disease, atherosclerosis, and septic shock [36
]. Activation of NF-B takes place in humans with unstable angina [37
], and it has been shown to play a role in ischemia-reperfusion injury [38
]. In addition, NF-B expression is elevated in regions of high probability for the development of arteriosclerotic plaques [39
]. Conversely, fatty, diet-fed mice deficient in NF-B signaling exhibit reduced fatty streak formation [40
], and a cis-element decoy against the NF-B binding site inhibits intimal hyperplasia following balloon injury in rat carotid artery [38
].
These observations, the presence of PTX3 during the early phases of inflammation and the enhancement of c-Rel/p65 activation by PTX3, offer the biological plausibility for a role of PTX3 in modulation of MN procoagulant activity.
In our previous study, we have shown that PTX3 could enhance TF by human umbilical vein endothelial cells exposed to LPS, IL-1ß, and TNF- [17
]. In contrast, no increase in TF could be observed in the present study, when IL-1ß and TNF- were used as inducing agents, suggesting a specificity for PTX3 toward the TF-inducing agent in MN. A possible explanation for the discordant behavior observed in the two cell types may reside in a different mechanism of regulation of TF synthesis by the different inducers. These results suggest that binding of an agonist to its receptor causes IB phosphorylation and degradation through different signal-transduction pathways depending on the nature of the agonist/receptor pair and on the cell type. The hypothesis of differential regulation of IB through the existence of cell-specific signaling pathways has recently been proposed by Holschermann et al. [41
], who demonstrated that the fungal-derived immunosuppressive drug cyclosporin A inhibits TF expression in monocytes but enhances it in endothelial cells. In their paper, Holschermann et al. [41
] suggest that the phospatase calcineurin may be the target enzyme of this action: calcineurin differently modulates NF-B transactivation at the level of IB by enhancing, in monocytes, or suppressing, in endothelial cells, IB phosphorylation.
In conclusion, the present data, together with those of our previous report about endothelial cells, offer two pieces of evidence for the complex puzzle of inflammation/coagulation interactions. During the onset of inflammation, molecules inducing the procoagulant signal responsible for the local fibrin deposition are generated. Monocytes and endothelial cells, at sites of vascular injury and inflammation, actively participate in this general scheme expressing TF on their membrane. PTX3, once generated, potentiates the expression of TF, which is required for thrombogenesis and wound healing. These processes may be of particular relevance for the amplification of clotting activation on advanced atherosclerotic plaques, where PTX3 has been localized within monocytes and endothelial cells.
Received October 31, 2003; revised March 8, 2004; accepted March 9, 2004.
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
B/Rel, and Sp1 proteins in uninduced and lipopolysaccharide-induced expression Arterioscler. Thromb. Vasc. Biol. 17,365-374B phosphorylation and endothelial cell adhesion molecule expression show anti-inflammatory effects in vivo J. Biol. Chem. 272,21096-21103
and lipopolysaccharide potentiate monocyte tissue factor induction by C-reactive protein: relationship with age, sex, and hormone replacement treatment Circulation 101,1785-1791B and IB: an inducible regulatory system in endothelial activation J. Exp. Med. 179,503-512-B and the heart J. Am. Coll. Cardiol. 38,307-314B is selectively and markedly activated in humans with unstable angina pectoris Circulation 98,1707-1713B binding site: evidence for a role of NFB in ischemia-reperfusion injury Circulation 96,280-285 B signal transduction pathway in aortic endothelial cells is primed for activation in regions predisposed to atherosclerotic lesion formation Proc. Natl. Acad. Sci. USA 97,9052-9057This article has been cited by other articles:
 |
|
 |
|
  N. S. Jenny, A. M. Arnold, L. H. Kuller, R. P. Tracy, and B. M. Psaty Associations of Pentraxin 3 With Cardiovascular Disease and All-Cause Death: The Cardiovascular Health Study Arterioscler. Thromb. Vasc. Biol., April 1, 2009; 29(4): 594 - 599. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Loeffler, Z. Haddad, M. Bonin, N. Romeike, M. Mezger, U. Schumacher, M. Kapp, F. Gebhardt, G.-U. Grigoleit, S. Stevanovic, et al. Interaction analyses of human monocytes co-cultured with different forms of Aspergillus fumigatus J. Med. Microbiol., January 1, 2009; 58(1): 49 - 58. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Mallat and A. Tedgui HDL, PTX3, and Vascular Protection Arterioscler. Thromb. Vasc. Biol., May 1, 2008; 28(5): 809 - 811. [Full Text] [PDF]
|
|
|
|
 |
|
 M. Tong, J. J. Carrero, A. R. Qureshi, B. Anderstam, O. Heimburger, P. Barany, J. Axelsson, A. Alvestrand, P. Stenvinkel, B. Lindholm, et al. Plasma Pentraxin 3 in Patients with Chronic Kidney Disease: Associations with Renal Function, Protein-Energy Wasting, Cardiovascular Disease, and Mortality Clin. J. Am. Soc. Nephrol., September 1, 2007; 2(5): 889 - 897. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. He, B. Han, and M. Liu Long pentraxin 3 in pulmonary infection and acute lung injury Am J Physiol Lung Cell Mol Physiol, May 1, 2007; 292(5): L1039 - L1049. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Okutani, B. Han, M. Mura, T. K. Waddell, S. Keshavjee, and M. Liu High-volume ventilation induces pentraxin 3 expression in multiple acute lung injury models in rats Am J Physiol Lung Cell Mol Physiol, January 1, 2007; 292(1): L144 - L153. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Bottazzi, A. Bastone, A. Doni, C. Garlanda, S. Valentino, L. Deban, V. Maina, A. Cotena, F. Moalli, L. Vago, et al. The long pentraxin PTX3 as a link among innate immunity, inflammation, and female fertility J. Leukoc. Biol., May 1, 2006; 79(5): 909 - 912. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Han, M. Mura, C. F. Andrade, D. Okutani, M. Lodyga, C. C. dos Santos, S. Keshavjee, M. Matthay, and M. Liu TNF{alpha}-Induced Long Pentraxin PTX3 Expression in Human Lung Epithelial Cells via JNK J. Immunol., December 15, 2005; 175(12): 8303 - 8311. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
