Roles of CD147 on T lymphocytes activation and MMP-9 secretion in systemic lupus erythematosus

Gina Pistol, Cristiana Matache, Ana Calugaru, Crina Stavaru, Stefanita Tanaseanu, Ruxandra Ionescu, Sergiu Dumitrache, Maria Stefanescu, Gina Pistol, Cristiana Matache, Ana Calugaru, Crina Stavaru, Stefanita Tanaseanu, Ruxandra Ionescu, Sergiu Dumitrache, Maria Stefanescu

Abstract

The cellular and molecular mechanisms involved in many abnormalities described in Systemic Lupus Erythematosus (SLE) are still unclear. Some of these abnormalities referred to the hyperactivation of T lymphocytes and the enhanced secretion of MMP-9 by peripheral blood mononuclear cells (PBMCs). Therefore, in this paper we investigated the potential role of CD147 molecule in these abnormalities. Our results demonstrated that CD147 molecule is overexpressed on CD3+T lymphocytes from SLE patients when compared with CD3+T lymphocytes from healthy donors. Monoclonal anti-CD147 antibodies, MEM-M6/1 clone, were able to inhibit protein tyrosine phosphorylation only in CD3 x CD28 costimulated T lymphocytes from SLE patients. However, this monoclonal antibody was unable to inhibit the enhanced activity of MMP-9 secreted by SLE PBMCs.

Figures

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1
FACS analysis of CD3+CD147+T lymphocytes. PBMCs freshly isolated from one subject were double stained using FITC conjugated monoclonal anti-CD147 antibodies (FL1-H) and PE conjugated monoclonal anti-CD3 antibodies (FL2-H). CD3+T lymphocytes were gated (A, gate R1). The percentage of CD3+CD147+T lymphocytes from R1 is identified in upper right quadrant (B).
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The percentage of CD147 positive cells in CD3+T lymphocytes from SLE patients and healthy donors. Points represent the percentages of CD3+CD147+T lymphocytes obtained for each active (aSLE) and inactive (iSLE) SLE patient and healthy donor (HD) as well as the mean values ± SD calculated for each group of subjects.
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3
FACS analysis for the density of CD147 molecules on CD3+T lymphocytes from SLE patients and healthy donors. Median of fluorescence intensities (MFI) that corresponds to the density of CD147 molecules on CD3+T lymphocytes is presented as points for each studied subject. The mean values ± SD of MFI for each group of subjects are included in histogram.
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Immunoblotting analysis of CD147 expression on PBMCs. PBMCs isolated from three healthy donors and three SLE patients (two in inactive and one in active stage of disease) were analyzed by immunoblotting using specific anti-CD147 and anti-actin antibodies. Immunoblotting was analyzed by densitometry and the expression level of CD147 on PBMCs was expressed as the ratio between CD147 and actin band intensities.
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Tyrosine phosphorylation pattern in unstimulated and stimulated T lymphocytes. PBMCs isolated from one SLE patient (A) and one healthy donor (B) were stimulated with different types of monoclonal antibodies, as follows: (1) unstimulated, (2) stimulated by anti-CD147 antibodies, (3) stimulated by anti-CD3 and anti-CD28 antibodies, (4) costimulated by antibodies directed to CD3, CD28 and CD147 receptors. After 5 min of stimulation, the cells were lysed and analyzed by immunoblotting with monoclonal anti-phosphotyrosine antibodies and ECL system. The standards of known molecular weight (MW) were included in each experiment.
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Tyrosine phosphorylation level in unstimulated and stimulated T lymphocytes. PBMCs isolated from SLE patients and healthy donors were stimulated as in Figure 5 and analyzed by densitometry for total tyrosine phosphorylation level. For each type of stimulation and for SLE (A) and healthy donors (B) group the mean values ± SD was calculated and indicated on histograms.
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Correlation between the percentage of CD3+CD147+T lymphocytes and the activity of secreted MMP-9. PBMCs freshly isolated from peripheral blood of SLE patients were analyzed by FACS in order to determine the percentage of CD3+CD147+T lymphocytes. In addition, PBMCs from the same patients were cultured for 24 hrs and the culture supernatants were analyzed by gelatin zymography. Zymograms were scanned and the intensity of bands was determined. Subsequently, these two parameters were correlated by Spearman's analysis.
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Modulation of MMPs activity in PBMCs cultured in absence or presence of monoclonal anti-CD147 antibodies. PBMCs of one SLE patient were cultured in absence (Line 2) or presence of anti-CD147 antibodies immobilized by rabbit anti-mouse IgG antibodies (Lines 4, 5) or soluble (lines 6, 7). Two controls were included in all experiments: positive control (fetal calf serum as source of latent and active forms of MMP-9 and latent MMP-2) (Line 1) and negative control (supernatant of PBMCs cultured only in the presence of rabbit antimouse IgG antibodies) (Line 3). After 24 hours, supernatants were collected and analyzed by gelatin zymography.

References

    1. Kammer GM, Perl A, Richardson BC, Tsokos GC. Abnormal T cell signal transduction in systemic lupus erythematosus. Arthritis Rheum. 2002;46:1139–54.
    1. Blasini AM, Brundula V, Paris M, Rivas L, Salazar S, Stekman IL, Rodriguez MA. Protein tyrosine kinase activity in T lymphocytes from patients with systemic lupus erythematosus. J Autoimmun. 1998;11:387–93.
    1. Matache C, Stefanescu M, Onu A, Tanaseanu S, Matei I, Frade R, Szegli G. p56lck activity and expression in peripheral blood lymphocytes from patients with systemic lupus erythematosus. Autoimmunity. 1999;29:111–20.
    1. Nambiar MP, Enyedy EJ, Fisher CU, Krishnan S, Warke VG, Gilliland WR, Oglesby RJ, Tsokos GC. Abnormal expression of various molecular forms and distribution of T cell receptor zeta chain in patients with systemic lupus erythematosus. Arthritis Rheum. 2002;46:163–74.
    1. Nambiar MP, Krishnan S, Warke VG, Tsokos GC. TCR zeta-chain abnormalities in human systemic lupus erythematosus. Methods Mol Med. 2004;102:49–72.
    1. Vassilopoulos D, Kovacs B, Tsokos GC. TCR/CD3 complex–mediated signal transduction pathway in T cells and T cell lines from patients with systemic lupus erythematosus. J Immunol. 1995;155:2269–81.
    1. Takeuchi T, Pang M, Amano K, Koide, Abe T. Reduced protein tyrosine phosphatase (PTPase) activity of CD45 on peripheral blood lymphocytes in patients with systemic lupus erythematosus (SLE) Clin Exp Immunol. 1997;109:20–6.
    1. Theofilopoulos AN. Immune complexes in autoimmunity. In: Bona CA, Siminovitch KA, Zanetti M, Theofilopoulos AN, editors. The molecular pathology of autoimmune diseases. Switzerland: Harwood: Academic Publishers; 1993. pp. 229–44.
    1. Matache C, Stefanescu M, Dragomir C, Tanaseanu S, Onu A, Ofiteru A, Szegli G. Matrix metalloproteinase- 9 and its natural inhibitor TIMP-1 expressed or secreted by peripheral blood mononuclear cells from patients with systemic lupus erythematosus. J Autoimmun. 2003;20:323–31.
    1. Kasinrerk W, Fiebiger E, Stefanova I, Baumruker T, Knapp W, Stockinger H. Human leukocyte activation antigen M6, a member of the Ig superfamily, is the species homologue of rat OX-47, mouse basigin, and chicken HT7 molecule. J Immunol. 1992;149:847–54.
    1. Biswas C, Zhang Y, DeCastro R, Guo H, Nakamura T, Kataoka H, Nabeshima K. The human tumor cellderived collagenase stimulatory factor (renamed EMMPRIN) is a member of the immunoglobulin superfamily. Cancer Res. 1995;55:434–9.
    1. Miyauchi T, Masuzawa Y, Muramatsu T. The basigin group of the immunoglobulin superfamily: complete conservation of a segment in and around transmembrane domains of human and mouse basigin and chicken HT7 antigen. J Biochem (Tokyo) 1991;110:770–4.
    1. Fossum S, Mallett S, Barclay AN. The MRC OX-47 antigen is a member of the immunoglobulin superfamily with an unusual transmembrane sequence. Eur J Immunol. 1991;21:671–9.
    1. DeCastro R, Zhang Y, Guo H, Kataoka H, Gordon MK, Toole B, Biswas G. Human keratinocytes express EMMPRIN, an extracellular matrix metalloproteinase inducer. J Invest Dermatol. 1996;106:1260–5.
    1. Nehme CL, Fayos BE, Bartles JR. Distribution of the integral plasma membrane glycoprotein CE9 (MRC OX-47) among rat tissues and its induction by diverse stimuli of metabolic activation. Biochem J. 1995;310:693–8.
    1. Koch C, Staffler G, Huttinger R, Hilgert I, Prager E, Cerny J, Steinlein P, Majdic O, Horejsi V, Stockinger H. T cell activation-associated epitopes of CD147 in regulation of the T cell response, and their definition by antibody affinity and antigen density. Int Immunol. 1999;11:777–86.
    1. Renno T, Wilson A, Dunkel C, Coste I, Maisnier-Patin K, Benoit de Coignac A, Aubry JP, Lees RK, Bonnefoy JY, MacDonald HR, Gauchat JF. A role for CD147 in thymic development. J Immunol. 2002;168:4946–50.
    1. Stonehouse TJ, Woodhead VE, Herridge PS, Ashrafian H, George M, Chain BM, Katz DR. Molecular characterization of U937-dependent T-cell co-stimulation. Immunology. 1999;96:35–47.
    1. Woodhead VE, Stonehouse TJ, Binks MH, Speidel K, Fox DA, Gaya A, Hardie D, Henniker AJ, Horejsi V, Sagawa K, Skubitz KM, Taskov H, Todd RF, 3rd, Van Agthoven A, Katz DR, Chain BM. Novel molecular mechanisms of dendritic cell-induced T cell activation. Int Immunol. 2000;12:1051–61.
    1. Zhou J, Zhu P, Jiang J, Jiang Q, Wu ZB, Yao XY, Thang H, Lu N, Yang Y, Chen ZN. Involvement of CD147 in overexpression of MMP-2 and MMP-9 and enhancement of invasise potential of PMA-differentiated THP-1. Cell Biology. 2005;6:1–10.
    1. Stockinger H, Ebel T, Hansmann C, Koch C, Majdic O, Prager E, Patel DD, Fox DA, Horejsi V, Sagawa K, Shen D-C. CD147 (neurothelin/basigin) Workshop Panel Report. In: Kishimoto T, Kikutani H, Von Dem Borne AEGK, Goyert SM, Mason DY, Miyasaka M, Moretta L, Okumura K, Shaw S, Springer TA, Sugamura K, Zola H, editors. Leucocyte Typing VI. New York: Garland Publishing, Inc; 1997. pp. 760–5.
    1. Gabison EE, Hoang-Xuan T, Mauviel A, Menashi S. EMMPRIN/CD147, an MMP modulator in cancer, development and tissue repair. Biochimie. 2005;87:361–8.
    1. Zucker S, Hymowitz M, Rollo EE, Mann R, Conner CE, Cao J, Foda HD, Tompkins DC, Toole BP. Tumorigenic potential of extracellular matrix metalloproteinase inducer. Am J Pathol. 2001;158:1921–8.
    1. Caudroy S, Polette M, Nawrocki-Raby B, Cao J, Toole BP, Zucker S, Birembaut P. EMMPRIN mediated MMP regulation in tumor and endothelial cells. Clin Exp Metastasis. 2002;19:697–702.
    1. Yurchenko V, Zybarth G, O'Connor M, Dai WW, Franchin G, Hao T, Guo H, Hung HC, Toole B, Gallay P, Sherry B, Bukrinsky M. Active site residues of cyclophilin A are crucial for its signaling activity via CD147. J Biol Chem. 2002;277:22959–65.
    1. Allain F, Vanpouille C, Carpentier M, Slomianny MC, Durieux S, Spik G. Interaction with glycosaminoglycans is required for cyclophilin B to trigger integrinmediated adhesion of peripheral blood T lymphocytes to extracellular matrix. Proc Natl Acad Sci USA. 2002;99:2714–9.
    1. Pushkarsky T, Yurchenko V, Vanpouille C, Brichacek B, Vaisman I, Hatakeyama S, Nakayama KI, Sherry B, Bukrinsky MI. Cell surface expression of CD147/emmprin is regulated by cyclophilin 60. J Biol Chem. 2005;280:27866–71.
    1. Zhou S, Zhou H, Walian PJ. CD147 is a regulatory subunit of the gamma-secretase complex in Alzheimer's disease amyloid beta-peptide production. Proc Natl Acad Sci USA. 2005;102:7499–504.
    1. Zhu P, Ding J, Zhou J, Dong WJ, Fan CM, Chen ZN. Expression of CD147 on monocytes/macrophages in rheumatoid arthritis: its potential role in monocyte accumulation and matrix metalloproteinase production. Arthritis Res Ther. 2005;7:1023–33.
    1. Zhu P, Lu N, Shi ZG, Zhou J, Wu ZB, Yang Y, Ding J, Chen ZN. CD147 overexpression on synoviocytes in rheumatoid arthritis enhances matrix metalloproteinase production and invasiveness of synoviocytes. Arthritis Res Ther. 2006;8:1–12.
    1. Yang JM, O'Neill P, Jin W, Foty R, Medina DJ, Xu Z, Lomas M, Arndt GM, Tang Y, Nakada M, Yan L, Hait WN. Extracellular matrix metalloproteinase inducer (CD147) confers resistance of breast cancer cells to Anoikis through inhibition of Bim. J Biol Chem. 2006;281:9719–27.
    1. Hochberg MC. Updating the American College of Rheumatology revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum. 1997;40:1725–35.
    1. Staffler G, Szekeres A, Schutz GJ, Saemann MD, Prager E, Zeyda M, Drbal K, Zlabinger GJ, Stulnig TM, Stockinger H. Selective inhibition of T cell activation via CD147 through novel modulation of lipid rafts. J Immunol. 2003;171:1707–14.
    1. Igakura T, Kadomatsu K, Taguchi O, Muramatsu H, Kaname T, Miyauchi T, Yamamura K, Arimura K, Muramatsu T. Roles of basigin, a member of the immunoglobulin superfamily, in behavior as to an irritating odor, lymphocyte response, and blood-brain barrier. Biochem Biophys Res Commun. 1996;224:33–6.
    1. Nabeshima K, Suzumiya J, Nagano M, Ohshima K, Toole BP, Tamura K, Iwasaki H, Kikuchi M. Emmprin, a cell surface inducer of matrix metalloproteinases (MMPs), is expressed in T-cell lymphomas. J Pathol. 2004;202:341–51.
    1. Konttinen YT, Li TF, Mandelin J, Liljestrom M, Sorsa T, Santavirta S, Virtanen I. Increased expression of extracellular matrix metalloproteinase inducer in rheumatoid synovium. Arthritis Rheum. 2000;43:275–80.
    1. Schmidt R, Bultmann A, Ungerer M, Joghetaei N, Bulbul O, Thieme S, Chavakis T, Toole BP, Gawaz M, Schomig A, May AE. Extracellular matrix metalloproteinase inducer regulates matrix metalloproteinase activity in cardiovascular cells: implications in acute myocardial infarction. Circulation. 2006;113:834–41.
    1. Abe N, Osanai T, Fujiwara T, Kameda K, Matsunaga T, Okumura K. C-reactive protein-induced upregulation of extracellular matrix metalloproteinase inducer in macrophages: inhibitory effect of fluvastatin. Life Sci. 2006;78:1021–8.
    1. Chiampanichayakul S, Peng-in P, Khunkaewla P, Stockinger H, Kasinrerk W. CD147 contains different bioactive epitopes involving the regulation of cell adhesion and lymphocyte activation. Immunobiology. 2006;211:167–78.
    1. Jury EC, Kabouridis PS, Flores-Borja F, Mageed RA, Isenberg DA. Altered lipid raft–associated signaling and ganglioside expression in T lymphocytes from patients with systemic lupus erythematosus. J Clin Invest. 2004;113:1176–87.

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