Objective: To establish the relationship between T cell responses to integrin coreceptor stimulation and B cell hyperreactivity as measured by pathologic autoantibody production.
Methods: Peripheral blood mononuclear cells from 42 patients with SLE according to the American Rheumatism Association criteria were examined for their ability to adhere to plateimmobilised fibronectin. Co-stimulation assays were performed on the same cells using anti-CD3 antibody alone or co-immobilised with an anti-b1-integrin antibody. Proliferative responses were measured by 3[H]thymidine pulsing on day 3 and activation was determined using a commercial protein kinase C assay, the protocol being established by our group in association with Promega. b1-Integrin expression was established by FACS analysis.
Results: An impaired PKC response to integrin-mediated activation was found in T-lymphocytes from 6=21 (29%) SLE patients, which correlated significantly with an absence of anti-dsDNA antibody in patient sera, irrespective of prednisolone treatment. Integrin co-stimulation of TcR/CD3-induced proliferation and T cell adhesion to fibronectin were also impaired among 5=21 (24%) and 6=15 (40%) patients studied, respectively.
Conclusion: We hypothesise that the integrity of b1-integrin signalling pathways may influence pathological antibody production in SLE by affecting T-lymphocyte activation and interactions between T and Blymphocytes.
Mohan C , Shi Y , Laman JD , Datta SK . Interaction between CD40 and its ligand gp39 in the development of murine lupus nephritis . J Immunol1995; 154: 1470–1480 .
3.
Finck BK , Linsley PS , Wofsy D . Treatment of murine lupus with CTLA4Ig . Science1994; 265: 1225–1227 .
4.
Mohan C , Adams S , Stanik V , Datta SK . Nucleosome: a major immunogen for pathogenic autoantibody-inducing T cells of lupus . J Exp Med1993; 177: 1367–1381 .
5.
Adams S , Leblanc P , Datta SK . Junctional region sequences of T-cell receptor beta-chain genes expressed by pathogenic anti-DNA autoantibody-inducing helper T cells from lupus mice: possible selection by cationic autoantigens . Proc Natl Acad Sci USA1991; 88: 11271–11275 .
6.
Rajagopalan S , Zordan T , Tsokos GC , Datta SK . Pathogenic anti-DNA autoantibody-inducing T helper cell lines from patients with active lupus nephritis: isolation of CD47 CD87 T helper cell lines that express the gd T-cell antigen receptor . Proc Natl Acad Sci USA1990; 87: 7020–7024 .
7.
Gallo CD , et al. Autoreactive kidney-infiltrating T-cell clones in murine lupus nephritis . Kidney Int1992; 42: 851–859 .
Hynes RO . Integrins: versatility, modulation, and signaling in cell adhesion . Cell1992; 69: 11–25 .
10.
Takeuchi T , et al. Upregulated expression and function of integrin adhesive receptors in systemic lupus erythematosis patients with vasculitis . J Clin Invest1993; 92: 3008–3016 .
11.
Shikata K , et al. Distribution of extracellular matrix receptors in various forms of glomerulonephritis . Am J Kidney Dis1995; 25: 680–688 .
Wuthrich RP , et al. Intercellular adhesion molecule-1 (ICAM-1) is upregulated in autoimmune lupus nephritis . Am J Pathol1990; 136: 441–450 .
14.
Yurochko AD , Liu DY , Eierman D , Haskill S . Integrins as a primary signal transduction molecule regulating monocyte immediate-early gene induction . Proc Natl Acad Sci1992; 89: 9034–9038 .
15.
Eierman DF , Johnson CE , Haskill JS . Human monocyte inflammatory mediator gene expression is selectively regulated by adherence substrates . J Immunol1989; 142: 1970–1976 .
16.
Matsushima K , et al. Molecular cloning of a human monocyte-derived neutrophil chemotactic factor (MDNCF) and the induction of MDNCF mRNA by interleukin-1 and tumor necrosis factor . J Exp Med1988; 167: 1883–1893 .
17.
Strieter RM , et al. Endothelial cell gene expression of a neutrophil chemotactic factor by TNF-a, LPS, and IL-1b . Science1989; 243: 1467–1469 .
18.
Iademarco MF , Barks JL , Dean DC . Regulation of vascular cell adhesion molecule-1 expression by IL-4 and TNF-a in cultured endothelial cells . J Clin Invest1995; 95: 264–271 .
19.
Swerlick RA , et al. Regulation of vascular cell adhesion molecule 1 on human dermal microvascular endothelial cells . J Immunol1992; 149: 698–705 .
20.
Huhtala P , et al. Cooperative signaling by a5=b1 and a4=b1 integrins regulates metalloproteinase gene expression in fibroblasts adhering to fibronectin . J Cell Biol1995; 129: 867–879 .
21.
Riikonen T , et al. Integrin a2b1 is a positive regulator of collagenase (MMP-1) and collagen a1(I) gene expression . J Biol Chem1995; 270: 13548–13552 .
22.
Vincenti MP , Clark IM , Brinckerhoff CE . Using inhibitors of metalloproteinases to treat arthritis . Arthritis Rheum1994; 37: 1115–1126 .
23.
Via CS , et al. T cell–antigen-presenting cell interactions in human systemic lupus erythematosus . J Immunol1993; 151: 3914–3922 .
24.
Tan EM , et al. The 1982 revised criteria for the classification of systemic lupus erythematosus . Arthritis Rheum1982; 25: 1271–1277 .
25.
Vitali C , et al. Disease activity in systemic lupus erythematosus: report of the Consensus Study Group of the European Workshop for Rheumatology Research . Clin Exp Rheum atol1992; 10: 541–547 .
26.
Mould AP , et al. Identification of a novel anti-integrin monoclonal antibody that recognises a ligand-induced binding site epitope on the b1 subunit . FEBS Lett1995; 363: 118–122 .
27.
Kinchington D , Ng T. Separation and maintenance of primary T and B lymphocytes . Methods Mol Biol1997; 75: 91–100 .
28.
Ager A , Humphries MJ . Use of synthetic peptides to probe lymphocyte–high endothelial cell interactions. Lymphocytes recognize a ligand on the endothelial surface which contains the CS1 adhesion motif . Int Immunol1990; 2: 921–928 .
29.
Nojima Y , et al. Ligation of VLA-4 on T cells stimulates tyrosine phsophorylation of a 105kD protein . J Exp Med1992; 175: 1045–1053 .
30.
Ng TTC , et al. Adhesion co-receptor expression and intracellular signalling in HIV disease: implications for immunotherapy . AIDS1995; 9: 337–343 .
31.
Pomies P , Frachet P , Block MR . Control of the a5b1 integrin/fibronectin interaction in vitro by the serine/threonine protein phosphatase calcineurin . Biochemistry1995; 34: 5104–5112 .
32.
Linker-Israeli M , et al. Defective production of interleukin 1 and interleukin 2 in patients with systemic lupus erythematosus (SLE) . J Immunol1983; 130: 2651–2655 .
33.
Alcocer-Varela J , Alarcon-Segovia D . Decreased production of and response to interleukin-2 by cultured lymphocytes from patients with systemic lupus erythematosus . J Clin Invest1982; 69: 1388–1392 .
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 Immunol1995; 155: 2269–2281 .
40.
Sfikakis PP , Oglesby R , Sfikakis P , Tsokos GC . B7=BB1 provides an important costimulatory signal for CD3-mediated T lymphocyte proliferation in patients with systemic lupus erythematosus (SLE) . Clin Exp Immunol1994; 96: 8–14 .
41.
Berry N , Nishizuka Y . Protein kinase C and T cell activation . Eur J Biochem1990; 189: 205–214 .
42.
Genot EM , Parker PJ , Cantrell DA . Analysis of the role of protein kinase C-a,-e, and z in T cell activation . J Biol Chem1995; 270: 9833–9839 .
43.
Illera VA , et al. Apoptosis in splenic B lymphocytes. Regulation by protein kinase C and IL-4 . J Immunol1993; 151: 2965–2973 .
44.
Evans CA , et al. Suppression of apoptosis by v-ABL protein tyrosine kinase is associated with nuclear translocation and activation of protein kinase C in an interleukin-3-dependent haemopoietic cell line . J Cell Sci1995; 108: 2591–2598 .
45.
McConkey DJ , Orrenius S , Jondal M . Cellular signalling in programmed cell death (apoptosis) . Immunol Today1990; 11: 120–121 .
46.
Chong P , et al.. Inhibition of protein-kinase C in peripheral blood mononuclear cells of patients with systemic lupus erythematosus: effect of spontaneous immunoglobulin production . Autoimmunity1991; 10: 227–231 .
47.
Boudreau N , Sympson CJ , Werb Z , Bissell MJ . Suppression of ICE and apoptosis in mammary epithelial cells by extracellular matrix . Science1995; 267: 891–893 .
48.
Meredith JE , Fazeli B , Schwartz MA . The extracellular matrix as a cell survival factor . Mol Biol Cell1993; 4: 953–961 .
49.
Zhang Z , Vuori K , Reed JC , Ruoslahti E . The a5b1 integrin supports survival of cells on fibronectin and up-regulates Bcl-2 expression . Proc Natl Acad Sci1995; 92: 6161–6165 .
50.
Hoyt DG , et al. Integrin activation suppresses etoposide-induced DNA strand breakage in cultured murine tumor-derived endothelial cells . Cancer Res1996; 56: 4146–4149 .
51.
Rich S , Nood NV , Lee HM . Role of a5b1 integrin in TGF b1costimulated CD8/ T cell growth and apoptosis . J Immunol1996; 157: 2916–2923 .
52.
Emlen W , Niebur J , Kadera R . Accelerated in vitro apoptosis of lymphocytes from patients with systemic lupus erythematosus . J Immunol1994; 152: 3685–3692 .
53.
Ohkusu K , Isobe K , Hidaka H , Nakashima I . Elucidation of the protein kinase C-dependent apoptosis pathway in distinct subsets of T lymphocytes in MRL-lpr/lpr mice . Eur J Immunol1995; 25: 3180–3186 .
54.
McNamee HP , Ingber DE , Schwartz MA . Adhesion to fibronectin stimulates inositol lipid synthesis and enhances PDGF-induced inositol lipid breakdown . J Cell Biol1993; 121: 673–678 .
55.
Auer KL , Jacobson BS . b1 integrins signal lipid second messengers required during cell adhesion . Mol Biol Cell1995; 6: 1305–1313 .
56.
Cybulsky AV , et al. Extracellular matrix-stimulated phospholipase activation is mediated by b1-integrin . Am J Physiol1993; 264: C323–C332 .
57.
Clark EA , Hynes RO . Ras activation is necessary for integrin-mediated activation of extracellular signal-regulated kinase 2 and cytosolic phospholipase A2 but not for cytoskeletal organization . J Biol Chem1996; 271: 14814–14818 .
58.
Cazaubon S , Bornancin F , Parker PJ . Threonine-497 is a critical site for permissive activation of protein kinase Ca . Biochem J1994; 301: 443–448 .
59.
Cazaubon SM , Parker PJ . Identification of the phosphorylated region responsible for the permissive activation of protein kinase C . J Biol Chem1993; 268: 17559–17563 .
60.
Bornancin F , Parker PJ . Phosphorylation of protein kinase C-a on serine 657 controls the accumulation of active enzyme and contributes to its phosphatase resistant state . J Biol Chem1997; 272: 3544–3549 .
61.
Bornancin F , Parker PJ . Phosphorylation of threonine 638 critically controls the dephosphorylation and inactivation of protein kinase C-a . Curr Biol1996; 6: 1114–1123 .
62.
Keranen LM , Dutil EM , Newton AC . Protein kinase C is regulated in vivo by three functionally distinct phosphorylations . Curr Biol1995; 5: 1394–1403 .
63.
Dutil EM , Keranen LM , DePaoli-Roach AA , Newton AC . In vivo regulation of protein kinase C by trans-phosphorylation followed by autophosphorylation . J Biol Chem1994; 269: 29359–29362 .
64.
Newton AC . Protein kinase C: structure, function and regulation . J Biol Chem1995; 270: 28495–28498 .
65.
Matsuzawa S , et al. Increase in potential activities of protein phosphatases PP1 and PP2A in lymphoid tissues of autoimmune MRL/MpJ-lpr/lpr mice . J Biochem1992; 111: 472–478 .
66.
Gordon C , et al. Active systemic lupus erythematosus is associated with the recruitment of naive/resting T-cells . Br J Rheumatol1996; 35: 226–230 .
