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IMMUNOBIOLOGY
From the Tumor Immunology Laboratory and Cancer
Immunotherapy and Gene Therapy Program, Istituto Scientifico H S. Raffaele and Vita-Salute San Raffaele University; Department of
Biology, University of Milan and Istituto di Ricerche Farmacologiche
"Mario Negri," Milan, Italy; and the Sezione di Patologia e
Immunologia, Dipartimento di Biotecnologie, Università di
Brescia, Brescia, Italy.
Pentraxins are acute-phase proteins produced in vivo during
inflammatory reactions. Classical short pentraxins, C-reactive protein, and serum amyloid P component are generated in the
liver in response to interleukin (IL)-6. The long pentraxin PTX3 is produced in tissues under the control of primary proinflammatory signals, such as lipopolysaccharide, IL-1 Dying cells are removed from living tissues. Errors
in the clearance of cell corpses may cause
autoimmunity.1,2 Specific proteases (caspases) cleave many
autoantigens and determine the generation of nucleosomes, a major
antigen in the prototypic systemic autoimmune disease, systemic lupus
erythematosus (SLE).3 Key autoantigens cluster into
apoptotic blebs, and cleavage by granzyme B, which leads
cytotoxic T-cell-induced apoptosis, selectively modifies
autoantigens.4 Conversely, the processing of the
internalized dying cells yields T-cell epitopes.5
Scavenger phagocytes, which are devoid of the ability to initiate
immune responses, perform most of the clearance of dying cells in
vivo.1 However, the most potent antigen-presenting cells
(APCs), dendritic cells (DCs),6,7 also internalize dying
cells and present epitopes derived from their processing to major
histocompatibility complex (MHC) class I- and class II-restricted T
lymphocytes.8-15
DCs originate from bone marrow CD34+
progenitors.16 DC precursors enter the blood and reach
peripheral organs, where they develop to immature DCs. Immature DCs
capture soluble antigens via macropinocytosis7,16-18 and
particulates through phagocytosis.19-21 Primary
proinflammatory signals, such as interleukin-1 The outcome of presentation of antigens from dying cells by DCs
is tightly regulated. Cells (by definition, reservoirs of "self-antigens") die continuously during development and normal tissue turnover, and DCs constitutively shuttle the material derived from their intracellular processing to lymphoid organs.26
Normal cell death via apoptosis occurs in the absence of inflammatory signals causing DC maturation. Besides, apoptotic cell clearance by
macrophages triggers the release of anti-inflammatory
cytokines,27,28 including IL-10 and transforming growth
factor- In contrast, during inflammation, cells die in a context in which
proinflammatory cytokines promote DC maturation and T-cell priming and
immune responses start toward the initiating noxa (pathogens, etc). Immature DCs challenged with cells undergoing postapoptotic necrosis9,21 or directly killed by
necrosis11,13 mature and cross-present antigens to
autoreactive T cells. Therefore, dying cells and maturing DCs coexist,
threatening tolerance toward peripheral antigens.12
Autoimmunity, however, does not as a rule associate with inflammation.
Factors in the milieu, possibly recruited by the inflammatory signals
themselves, must prevent phagocytosis of dying cell by DCs.
Pentraxins are acute-phase proteins, usually characterized by cyclic
pentameric structure, that are conserved during
phylogenesis.31-33 Short pentraxins are produced in the
liver in response to inflammatory mediators.34,35 They
recognize several ligands, including substrates targeted during
systemic autoimmunity such as chromatin or small nuclear
ribonucleoproteins.36-40 The function of pentraxins
includes amplification of innate resistance against microbes and
regulation of the scavenging of DNA released from dying
cells.32
PTX3 is the prototypic long pentraxin, structurally related to,
but distinct from, C-reactive protein (CRP) and serum amyloid P
component (SAP). Signals, including lipopolysaccharide (LPS), IL-1 In this study, we report that PTX3 binds to dying but not to living
cells and that recognition is restricted to a phase in which nuclear
antigens segregate to the cell membrane. The binding to apoptotic cells
during inflammatory reactions and the regulation of clearance by DCs
may contribute to limit autoimmune phenomena.
Cells
Apoptosis induction and detection
Proteins Human PTX3 was purified from Chinese hamster ovary (CHO) cells stably and constitutively expressing the protein, as described.45 Purified human CRP, SAP, and bovine serum albumin (BSA) were purchased from Sigma (St Louis, MO). Biotinylation was performed as described.45 Biotinylated PTX3 was analyzed in the native state in 5% to 10% gradient polyacrylamide gel electrophoresis (PAGE) as described.45 Gels were stained with silver nitrate. Molecular weight markers for native gels (Pharmacia Biotech Europe, Brussels, Belgium) are as follows: thyroglobulin, 669 000; ferritin, 440 000; catalase, 232 000; lactate dehydrogenase, 140 000; BSA, 67 000. Human 2-glycoprotein I (2-GPI) was purified from normal human serum by sequential perchloric acid precipitation, heparin-sepharose (HiTrap Heparin, Pharmacia) affinity chromatography and cationic exchange chromatography (Resource S, Pharmacia), as described.48
Binding of soluble factors to apoptotic cells Cells were challenged with biotinylated PTX3 (range tested 0.1 to 500 µg/mL) for 30 minutes at room temperature. Cell-bound cofactors were revealed by flow cytometry after addition of FITC-conjugated streptavidin (Pierce, Rockford, IL). The fluorescence background was calculated with FITC-conjugated streptavidin only. The binding of purified2-GPI (10 µg/mL) was revealed by means of
rabbit polyclonal anti-2-GPI antibodies and FITC-labeled antirabbit antibodies (Sigma).48 Propidium iodide (PI) (10 µg/mL)
was added immediately before flow cytometry. When indicated, assays
were performed in the absence of divalent cations. The specificity of
the binding was verified before incubating apoptotic cells with PTX3,
SAP, CRP, and BSA (500 µg/mL) and before addition of biotinylated
PTX3 (10 µg/mL) and FITC streptavidin, and fluorescence-activated cell sorter (FACS) analysis. The inhibition percentage was calculated as follows: 100  (mean fluorescence intensity in the presence of
biotinylated PTX3 and FITC-streptavidin / mean fluorescence intensity
in the presence of FITC-streptavidin alone) × 100.
Phagocytosis Jurkat cells were committed to apoptosis by UV irradiation and labeled with the green fluorescent aliphatic dye PKH2-GL (Sigma).5,49 After washing, cells were incubated or not with PTX3 (100 µg/mL for 30 minutes at room temperature) and co-incubated with DCs for 90 minutes at 37°C or 4°C. Both immature DCs or DCs that matured after treatment with TNF- 20 were
used. As a control, the internalization of FITC-labeled ovalbumin
(Sigma) or of green fluorescent latex beads (2 µm diameter)
(Polysciences, Warrington, PA) by immature and mature DCs was assessed
in the presence or absence of PTX3. After phagocytosis, DCs were
incubated in PBS containing EDTA and trypsin for 15 minutes at 37°C
and analyzed by flow cytometry (FACS Scan, Becton Dickinson, San
Josè, CA). When indicated, trypsin was omitted and the
percentages of cells that bound or internalized apoptotic cells were
compared in independent samples.
DC maturation Jurkat cells were committed to apoptosis by UV irradiation or killed by primary necrosis (5 cycles of freezing and thawing). After washing, cells were incubated or not with PTX3, as described above, and coincubated with immature DCs for 24 hours at 37°C. DCs were also incubated in the presence of PTX3 alone. As a positive control, DCs were committed to maturation with the use of LPS (10 ng/mL). DCs were then retrieved by centrifugation over a Percol density gradient (50% to 30% interface). DCs were then analyzed by flow cytometry (FACS Scan) for MHC class II, CD40, CD80, and CD86 expression by means of appropriately diluted FITC- or phycoerythrin-labeled monoclonal antibodies (Caltag, Burlingame, CA).Confocal microscopy Stained cells were seeded on polylysine-coated coverslip for 20 minutes at room temperature, fixed for 15 minutes in 4% (wt/vol) paraformaldehyde, and mounted in Mowiol medium. Confocal laser scanning microscopy was performed with a Leica TCS-NT (Leica Microsystems, Heidelberg, Germany) confocal laser scanning microscope, equipped with laser Argon/Krypton, 75-mW multiline. Focal series of horizontal planes of section were simultaneously monitored for FITC and PI by means of the 488- and 568-nm laser lines, a FITC band pass 590/30 filter, and a long pass 590 filter for PI.
PTX3 binds to dying cells Jurkat cells were propagated in TCM-nut to avoid interference by uncharacterized serum cofactors and were committed to apoptosis by CD95 cross-linking or UV irradiation. After both treatments, most Jurkat T cells (more than 95%) underwent apoptosis, as verified by morphological and cytometric criteria.47 PTX3 was purified from CHO cells stably and constitutively expressing the protein and the binding to living or apoptotic cells revealed by flow cytometry. Biotinylated PTX3 did not bind to living Jurkat cells (Figure 1A). In contrast, it bound efficiently after induction of apoptosis (Figure 1B; Table 1). The procedure of biotinylation does not influence the structural or functional integrity of the molecule.45 Accordingly, purified biotinylated PTX3, PTX3 in the conditioned medium of transfected CHO cells, and purified PTX3 all migrated as 440-kd apparent molecular mass species, possibly corresponding to a decamer, when gel electrophoresis was performed in nondenaturing, nonreducing conditions (Figure 1C and Bottazzi et al45). Linear increase of the binding was observed when increasing concentrations of biotinylated PTX3 were used; it reached a plateau at around 100 µg/mL (Figure 1D). The preincubation of apoptotic cells with the native molecule inhibited the binding (Figure 1E), demonstrating its specificity. The preincubation with an excess of unlabeled CRP or SAP also inhibited PTX3 binding to apoptotic cells (Figure 1E); the preincubation with the unrelated plasma protein BSA did not exert any effect (Figure 1E). PTX3 does not have a specific site for Ca++ and does not bind classical Ca++-dependent ligands of pentraxins.45 Accordingly, the binding of PTX3 to dying cells did not require the presence of extracellular divalent cations (Figure 1F), which were necessary for binding of the unrelated plasma factor annexin V (Figure 1F).
PTX3 binds to membrane domains of apoptotic cells The binding of PTX3 to dying cells was characterized by confocal imaging. Jurkat cells undergoing CD95-triggered apoptosis were easily identified on the basis of nuclear characteristics (Figure 2B,C,F, red color), such as chromatin margination, condensation, and fragmentation. Biotinylated PTX3 bound to discrete membrane domains of apoptotic cells (Figure 2D,F, green color), as revealed with FITC-labeled streptavidin. The binding of streptavidin alone to Jurkat cells was negligible (Figure 2A,C, green color). PTX3 did not bind to living cells, whose nuclei displayed normal morphology (Figure 2, upper left quadrant of each panel).
PTX3 recognition is restricted to a window during the apoptotic program We analyzed whether the phase of apoptosis influenced PTX3 binding. Figure 3 shows that PTX3 did not bind 2 hours after the cross-linking of the CD95 receptor (panel A). At this time point, the PS-binding proteins annexin V and2-GPI bound to most cells (Figure 3B-C). PTX3 acquired the ability
to bind to cells undergoing later apoptosis: most cells that were
recognized by PTX3 after 4 hours of treatment failed to exclude PI
(Figure 3E). On the contrary, 2-GPI and annexin V bound to both
PI+ and PI dying cells (Figure 3F-G). PTX3
did not bind to polymorphonuclear leukocytes undergoing early phases of
apoptosis (annexin V+ and PI) (Figure
4A). They acquired this ability only at
later time points (72 hours) (Figure 4B). PTX3 also recognized normal
  T cells after cross-linking of the CD95 receptor or antigen
recognition by the T-cell receptor50 during established
apoptosis only (Figure 4C).
PTX3 lost the ability to bind to cells evolving toward a postapoptotic phase (Table 1). Table 1 also shows that recognition by PTX3 does not depend on the initiating stimuli used to trigger apoptosis. Some binding to cells killed in vitro by necrosis was also observed. The mere entry of PTX3 into cells after permeabilization with saponin was not sufficient for binding, suggesting that cell death is necessary to make proper ligand binding sites available (Table 1). Apoptotic cells bound by PTX3 escape internalization by DCs Soluble factors bound to apoptotic cells may provide new ligands or mask signals for phagocytes (discussed in Ren and Savill1). We verified the rate of internalization of PTX3-bound apoptotic cells by immature human DCs, derived from circulating monocytes cultured in the presence of recombinant GM-CSF and IL-4.17,18 This treatment allowed the generation of homogenous populations of immature DCs (Figure 5A). Figure 5B shows that internalization of apoptotic cells abated as a consequence of PTX3 binding. This was not due to a toxic effect on DCs, since the molecule did not influence the uptake of fluorescent ovalbumin by macropinocytosis or of inert particulate substrates such as latex beads (Figure 5; Table 2). Furthermore, PTX3 did not influence the binding of apoptotic cells to immature DCs (Table 2). As expected, the ability to bind and to internalize apoptotic cells was down-regulated during DC maturation-ensuing treatment with TNF-
(Table 2). Treatment with PTX3 did not influence the extent of the
phenomenon (Table 2). Furthermore, treatment with PTX3 did not affect
the maturation state of DCs challenged with either apoptotic or
necrotic cells (not shown).
The general objective of this study was to assess whether the long
pentraxin PTX3 binds to dying cells and whether it regulates the
clearance by professional APCs. We found that spontaneous (polymorphonuclear neutrophils) or induced (normal and
transformed T lymphocytes) apoptosis is associated with recognition by
PTX3. Expression of PTX3 binding sites occurs late in the apoptotic process, subsequent to exposure of PS recognized by annexin V and
PTX3 binds also to necrotic cells, although less efficiently. This suggests that caspase activation is dispensable. Accordingly, interference with enzyme activity by means of diffusible peptide caspase inhibitors does not prevent PTX3 binding to dying cells (P.R. et al, unpublished data, March 1999). Suitable candidates may become available for pentraxin recognition-ensuing plasma membrane disruption. Besides classical ligands, such as phosphoethanolamine and phosphocoline, CRP binds to small nuclear ribonucleoproteins and SAP to chromatin/nucleolar components.36,38,51 These intracellular moieties abandon their original location and redistribute to the plasma membrane during late apoptosis.3,4 They are therefore present in the districts and during the time frame associated with PTX3 recognition. This preferential redistribution could also explain why PTX3 recognizes apoptotic cells with higher efficiency than necrotic cells. Alternatively, alteration in the glycosylation pathways, which occurs during cell death, may contribute to their recognition by pentraxins.40 Further studies are under way to define a possible role for these molecules as binding sites for PTX3 on the membrane of dying cells. The DC system of APCs is the initiator and modulator of normal immune responses.6,7 Immature DCs internalize dying cells, process them, and present antigens to T cells.7 This apoptosis-dependent antigen presentation pathway contributes to immune tolerance: DCs from noninflamed tissues phagocytose cells dying by apoptosis during normal tissue turnover, move to the T-cell areas of lymph nodes, and, in the absence of interaction with T cells, die. Resident DCs in the lymph node phagocytose the dying migratory DCs and induce tolerance in, or regulate, autoreactive T cells.10,30 During inflammation, the balance is skewed toward the initiation of immune responses. DCs mature, express costimulatory signals, and, after arrival at the lymph node, stimulate T cells that in turn maintain their viability.10 This event is crucial for cross-priming of T cells specific for pathogens.10,14 Efficient censorship mechanisms must prevent DC activation of
autoreactive T cells after internalization of dying cells during inflammation. This is particularly so since DCs do not mature when
challenged with low numbers of apoptotic cells,9
representative of apoptotic cells they meet in vivo during normal
tissue turnover or development. In contrast, in the presence of an
excess of dying cells that evolve toward postapoptotic necrosis, or of
cells killed by primary necrosis, DCs mature, up-regulate costimulatory
signal expression, and enhance their ability to cross-present antigens and initiate immune responses. Inflamed tissues The results described here suggest that the first cloned long pentraxin
PTX3 controls the interaction between maturing DCs and dying cells.
PTX3 is expressed by diverse cell types, most prominently mononuclear
phagocytes and endothelial cells, in response to primary
proinflammatory signals such as LPS, IL-1 Classical short pentraxins, produced in the liver under the control of IL-6, may serve the same function systemically. SAP, the major acute-phase reactant in the mouse, binds to keratin bodies.52 After immunization with native chromatin, mice with targeted deletion of the SAP gene develop a syndrome that resembles SLE, a systemic inflammatory autoimmune disease characterized by the generation of high-affinity T-cell-dependent autoantibodies.53 Regulation of antigen degradation by SAP may be involved in these findings.54 The results presented here suggest the alternative complementary explanation that pentraxins represent a safeguard mechanism against autoimmunity by preventing apoptotic cell internalization by APCs. Development of PTX3 gene-targeted mice, currently under way, will be useful to dissect the relative role of different pentraxins and to put the "PTX3 censorship" model to a test.
G. Balestrieri and A. Tincani for the gift of
Submitted March 2, 2000; accepted September 13, 2000.
Supported by the Associazione Italiana per la Ricerca sul Cancro (AIRC); the MURST "Cofinanziamento 1998" and "Cofinanziamento 1999"; and the Ministero della Sanità (Progetto Finalizzato Biotecnologie and Ricerca Finalizzata 1999).
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Angelo Manfredi, H S Raffaele, via Olgettina 60, 20132 Milano, Italy; e-mail: a.manfredi{at}hsr.it.
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© 2000 by The American Society of Hematology.
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  Y. IWATA, A. YOSHIZAKI, F. OGAWA, K. KOMURA, T. HARA, E. MUROI, M. TAKENAKA, K. SHIMIZU, M. HASEGAWA, M. FUJIMOTO, et al. Increased Serum Pentraxin 3 in Patients with Systemic Sclerosis J Rheumatol, May 1, 2009; 36(5): 976 - 983. [Abstract] [Full Text] [PDF]
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 J.-F. Augusto, C. Onno, S. Blanchard, S. Dubuquoi, A. Mantovani, A. Chevailler, P. Jeannin, and J.-F. Subra Detection of anti-PTX3 autoantibodies in systemic lupus erythematosus Rheumatology, April 1, 2009; 48(4): 442 - 444. [Full Text] [PDF]
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 L. Deban, H. Jarva, M. J. Lehtinen, B. Bottazzi, A. Bastone, A. Doni, T. S. Jokiranta, A. Mantovani, and S. Meri Binding of the Long Pentraxin PTX3 to Factor H: Interacting Domains and Function in the Regulation of Complement Activation J. Immunol., December 15, 2008; 181(12): 8433 - 8440. [Abstract] [Full Text] [PDF]
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 H. Wan, C. G. van Helden-Meeuwsen, C. Garlanda, L. M. E. Leijten, V. Maina, N. A. Khan, H. A. Drexhage, A. Mantovani, R. Benner, and M. A. Versnel Chorionic gonadotropin up-regulates long pentraxin 3 expression in myeloid cells J. Leukoc. Biol., November 1, 2008; 84(5): 1346 - 1352. [Abstract] [Full Text] [PDF]
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 G E Grossmayer, L E Munoz, C K Weber, S Franz, R E Voll, P M Kern, J R Kalden, G Schett, M Herrmann, and U S Gaipl IgG autoantibodies bound to surfaces of necrotic cells and complement C4 comprise the phagocytosis promoting activity for necrotic cells of systemic lupus erythaematosus sera Ann Rheum Dis, November 1, 2008; 67(11): 1626 - 1632. [Abstract] [Full Text] [PDF]
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 A. Doni, G. Mantovani, C. Porta, J. Tuckermann, H. M. Reichardt, A. Kleiman, M. Sironi, L. Rubino, F. Pasqualini, M. Nebuloni, et al. Cell-specific Regulation of PTX3 by Glucocorticoid Hormones in Hematopoietic and Nonhematopoietic Cells J. Biol. Chem., October 31, 2008; 283(44): 29983 - 29992. [Abstract] [Full Text] [PDF]
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W. Xu, S. P. Berger, L. A. Trouw, H. C. de Boer, N. Schlagwein, C. Mutsaers, M. R. Daha, and C. van Kooten Properdin Binds to Late Apoptotic and Necrotic Cells Independently of C3b and Regulates Alternative Pathway Complement Activation J. Immunol., June 1, 2008; 180(11): 7613 - 7621. [Abstract] [Full Text] [PDF]
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 M.E. Suliman, A.R. Qureshi, J.J. Carrero, P. Barany, M.I. Yilmaz, S. Snaedal-Jonsdottir, A. Alvestrand, O. Heimburger, B. Lindholm, and P. Stenvinkel The long pentraxin PTX-3 in prevalent hemodialysis patients: associations with comorbidities and mortality QJM, May 1, 2008; 101(5): 397 - 405. [Abstract] [Full Text] [PDF]
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A. Inforzato, V. Rivieccio, A. P. Morreale, A. Bastone, A. Salustri, L. Scarchilli, A. Verdoliva, S. Vincenti, G. Gallo, C. Chiapparino, et al. Structural Characterization of PTX3 Disulfide Bond Network and Its Multimeric Status in Cumulus Matrix Organization J. Biol. Chem., April 11, 2008; 283(15): 10147 - 10161. [Abstract] [Full Text] [PDF]
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 R. Glynne-Jones and P. Hoskin In Reply J. Clin. Oncol., March 20, 2008; 26(9): 1563 - 1563. [Full Text] [PDF]
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 R. M. Popovici, M. S. Krause, J. Jauckus, A. Germeyer, I. S. Brum, C. Garlanda, T. Strowitzki, and M. von Wolff The Long Pentraxin PTX3 in Human Endometrium: Regulation by Steroids and Trophoblast Products Endocrinology, March 1, 2008; 149(3): 1136 - 1143. [Abstract] [Full Text] [PDF]
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 M. Salio, S. Chimenti, N. De Angelis, F. Molla, V. Maina, M. Nebuloni, F. Pasqualini, R. Latini, C. Garlanda, and A. Mantovani Cardioprotective Function of the Long Pentraxin PTX3 in Acute Myocardial Infarction Circulation, February 26, 2008; 117(8): 1055 - 1064. [Abstract] [Full Text] [PDF]
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 S. Tranguch, A. Chakrabarty, Y. Guo, H. Wang, and S. K Dey Maternal Pentraxin 3 Deficiency Compromises Implantation in Mice Biol Reprod, September 1, 2007; 77(3): 425 - 432. [Abstract] [Full Text] [PDF]
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 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]
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 S. Jaillon, G. Peri, Y. Delneste, I. Fremaux, A. Doni, F. Moalli, C. Garlanda, L. Romani, H. Gascan, S. Bellocchio, et al. The humoral pattern recognition receptor PTX3 is stored in neutrophil granules and localizes in extracellular traps J. Exp. Med., April 16, 2007; 204(4): 793 - 804. [Abstract] [Full Text] [PDF]
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J. Gamrekelashvili, C. Kruger, R. von Wasielewski, M. Hoffmann, K. M. Huster, D. H. Busch, M. P. Manns, F. Korangy, and T. F. Greten Necrotic Tumor Cell Death In Vivo Impairs Tumor-Specific Immune Responses J. Immunol., February 1, 2007; 178(3): 1573 - 1580. [Abstract] [Full Text] [PDF]
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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]
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 M McMahon, J Grossman, W Chen, and B H Hahn Inflammation and the pathogenesis of atherosclerosis in systemic lupus erythematosus Lupus, November 1, 2006; 15(11_suppl): 59 - 69. [Abstract] [PDF]
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P. Baruah, I. E. Dumitriu, G. Peri, V. Russo, A. Mantovani, A. A. Manfredi, and P. Rovere-Querini The tissue pentraxin PTX3 limits C1q-mediated complement activation and phagocytosis of apoptotic cells by dendritic cells J. Leukoc. Biol., July 1, 2006; 80(1): 87 - 95. [Abstract] [Full Text] [PDF]
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E. F. Vernon-Wilson, F. Aurade, and S. B. Brown CD31 promotes {beta}1 integrin-dependent engulfment of apoptotic Jurkat T lymphocytes opsonized for phagocytosis by fibronectin J. Leukoc. Biol., June 1, 2006; 79(6): 1260 - 1267. [Abstract] [Full Text] [PDF]
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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]
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M Bijl, E Reefman, G Horst, P C Limburg, and C G M Kallenberg Reduced uptake of apoptotic cells by macrophages in systemic lupus erythematosus: correlates with decreased serum levels of complement Ann Rheum Dis, January 1, 2006; 65(1): 57 - 63. [Abstract] [Full Text] [PDF]
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 P. Baruah, A. Propato, I. E. Dumitriu, P. Rovere-Querini, V. Russo, R. Fontana, D. Accapezzato, G. Peri, A. Mantovani, V. Barnaba, et al. The pattern recognition receptor PTX3 is recruited at the synapse between dying and dendritic cells, and edits the cross-presentation of self, viral, and tumor antigens Blood, January 1, 2006; 107(1): 151 - 158. [Abstract] [Full Text] [PDF]
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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]
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L. E. Munoz, U. S. Gaipl, S. Franz, A. Sheriff, R. E. Voll, J. R. Kalden, and M. Herrmann SLE--a disease of clearance deficiency? Rheumatology, September 1, 2005; 44(9): 1101 - 1107. [Abstract] [Full Text] [PDF]
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C. J. G. de Almeida, L. B. Chiarini, J. P. da Silva, P. M. R. e Silva, M. A. Martins, and R. Linden The cellular prion protein modulates phagocytosis and inflammatory response J. Leukoc. Biol., February 1, 2005; 77(2): 238 - 246. [Abstract] [Full Text] [PDF]
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R. Latini, A. P. Maggioni, G. Peri, L. Gonzini, D. Lucci, P. Mocarelli, L. Vago, F. Pasqualini, S. Signorini, D. Soldateschi, et al. Prognostic Significance of the Long Pentraxin PTX3 in Acute Myocardial Infarction Circulation, October 19, 2004; 110(16): 2349 - 2354. [Abstract] [Full Text] [PDF]
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M Bijl, H Bootsma, Y van der Geld, P C Limburg, C G M Kallenberg, and M H van Rijswijk Serum amyloid P component levels are not decreased in patients with systemic lupus erythematosus and do not rise during an acute phase reaction Ann Rheum Dis, July 1, 2004; 63(7): 831 - 835. [Abstract] [Full Text] [PDF]
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M. Rusnati, M. Camozzi, E. Moroni, B. Bottazzi, G. Peri, S. Indraccolo, A. Amadori, A. Mantovani, and M. Presta Selective recognition of fibroblast growth factor-2 by the long pentraxin PTX3 inhibits angiogenesis Blood, July 1, 2004; 104(1): 92 - 99. [Abstract] [Full Text] [PDF]
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 G. Brouckaert, M. Kalai, D. V. Krysko, X. Saelens, D. Vercammen, `M. Ndlovu, G. Haegeman, K. D'Herde, and P. Vandenabeele Phagocytosis of Necrotic Cells by Macrophages Is Phosphatidylserine Dependent and Does Not Induce Inflammatory Cytokine Production Mol. Biol. Cell, March 1, 2004; 15(3): 1089 - 1100. [Abstract] [Full Text] [PDF]
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S. P. Hart, K. M. Alexander, and I. Dransfield Immune Complexes Bind Preferentially to Fc{gamma}RIIA (CD32) on Apoptotic Neutrophils, Leading to Augmented Phagocytosis by Macrophages and Release of Proinflammatory Cytokines J. Immunol., February 1, 2004; 172(3): 1882 - 1887. [Abstract] [Full Text] [PDF]
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L. C. Kao, A. Germeyer, S. Tulac, S. Lobo, J. P. Yang, R. N. Taylor, K. Osteen, B. A. Lessey, and L. C. Giudice Expression Profiling of Endometrium from Women with Endometriosis Reveals Candidate Genes for Disease-Based Implantation Failure and Infertility Endocrinology, July 1, 2003; 144(7): 2870 - 2881. [Abstract] [Full Text] [PDF]
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 B. Davidson and B. J. Swalla A molecular analysis of ascidian metamorphosis reveals activation of an innate immune response Development, March 12, 2003; 129(20): 4739 - 4751. [Abstract] [Full Text] [PDF]
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S. P. Hart, C. Jackson, L. M. Kremmel, M. S. McNeill, H. Jersmann, K. M. Alexander, J. A. Ross, and I. Dransfield Specific Binding of an Antigen-Antibody Complex to Apoptotic Human Neutrophils Am. J. Pathol., March 1, 2003; 162(3): 1011 - 1018. [Abstract] [Full Text] [PDF]
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B. Bussolati, G. Peri, G. Salvidio, D. Verzola, A. Mantovani, and G. Camussi The Long Pentraxin Ptx3 Is Synthesized in IgA Glomerulonephritis and Activates Mesangial Cells J. Immunol., February 1, 2003; 170(3): 1466 - 1472. [Abstract] [Full Text] [PDF]
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 S. Varani, J. A. Elvin, C. Yan, J. DeMayo, F. J. DeMayo, H. F. Horton, M. C. Byrne, and M. M. Matzuk Knockout of Pentraxin 3, a Downstream Target of Growth Differentiation Factor-9, Causes Female Subfertility Mol. Endocrinol., June 1, 2002; 16(6): 1154 - 1167. [Abstract] [Full Text] [PDF]
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 L. Stuart and J. Hughes Apoptosis and autoimmunity Nephrol. Dial. Transplant., May 1, 2002; 17(5): 697 - 700. [Full Text] [PDF]
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 E. Napoleone, A. Di Santo, A. Bastone, G. Peri, A. Mantovani, G. de Gaetano, M. B. Donati, and R. Lorenzet Long Pentraxin PTX3 Upregulates Tissue Factor Expression in Human Endothelial Cells: A Novel Link Between Vascular Inflammation and Clotting Activation Arterioscler. Thromb. Vasc. Biol., May 1, 2002; 22(5): 782 - 787. [Abstract] [Full Text] [PDF]
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M. S. Rolph, S. Zimmer, B. Bottazzi, C. Garlanda, A. Mantovani, and G. K. Hansson Production of the Long Pentraxin PTX3 in Advanced Atherosclerotic Plaques Arterioscler. Thromb. Vasc. Biol., May 1, 2002; 22(5): e10 - 14. [Abstract] [Full Text] [PDF]
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D. G. Souza, A. C. Soares, V. Pinho, H. Torloni, L. F. L. Reis, M. T. Martins, and A. A. M. Dias Increased Mortality and Inflammation in Tumor Necrosis Factor-Stimulated Gene-14 Transgenic Mice after Ischemia and Reperfusion Injury Am. J. Pathol., May 1, 2002; 160(5): 1755 - 1765. [Abstract] [Full Text] [PDF]
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G. K. Hansson Immune Mechanisms in Atherosclerosis Arterioscler. Thromb. Vasc. Biol., December 1, 2001; 21(12): 1876 - 1890. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
)
or late apoptosis (PI+, 
) are reported as RFI (y-axis),
calculated by dividing the mean fluorescence intensity in the presence
of biotinylated PTX3 and FITC-streptavidin with that obtained in the
presence of FITC-streptavidin alone.
where dying cells, antigen-capturing immature DCs, and maturative stimuli for DCs coexist