CD45 autoantibodies mediate neutralization of activated T cells from lupus patients through anergy or apoptosis

Abstract

The objectives were to provide estimates of the prevalence of autoantibody (Ab) directed to CD45 in lupus patients, identify the target autoantigen(s), and determine the ability of such reactivity to mediate neutralization of T lymphocytes.

Sera from 64 patients were studied using 2 assays: Western blot and an ELISA with CD45 eluted from 3 cell lines as antigen (U937, Jurkat and Daudi). The role of carbohydrate specificity was investigated using enzyme digestion of blotted glycans, competition with sugars, and inhibition with lectins. Apoptosis was studied through annexin V binding, and cell cycle analysis using the propidium iodide method.

AutoAb to CD45 were detected in 16/64 sera (25%) by Western blot, and 21/32 sera (66%) found positive in the ELISA. CD45 purified from Daudi cells was identified in the ELISA, but not in the blot. AutoAb were of the IgM and the IgG isotypes, but not specific for a particular cell type or CD45 isoform: 2 dominant specificities were recognized, one against p180, and another against high MW isoforms. Neuraminidase-induced enhancement of reactivity, together with the inhibitory effect of N-acetyl galactosamine and Dolichos diflorus lectin suggest that the epitopes are carbohydrates. AutoAb which were specific for activated CD4 + T cells triggered the annexin V binding, and, in 2 of 4 cases, lymphocytes underwent apoptosis.

In conclusion, carbohydrate conformational epitopes may be important as target antigens, and some CD45 autoAb have the capacity to neutralize activated T cells through anergy or apoptosis.

Keywords

systemic lupus erythematosus autoantibody T cell CD45

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References

Shoenfeld Y , Isenberg D . The Mosaic of Autoimmunity. Amsterdam: Elsevier , 1989.

Morimoto C , Steinberg AD , Letvin NL et al. A defect of immunoregulatory T cell subsets in systemic lupus erythematosus patients demonstrated with 2H4 antibody . J Clin Invest 1987; 79: 762–768 .

Winfield JB , Mimura T , Fernsten PD . Autoantibodies to CD45 in systemic lupus erythematosus . Int J Clin Lab Res 1992; 21: 292–295 .

Mimura T , Fernsten P , Jarjour W , Winfield JB . Autoantibodies specific for different isoforms of CD45 in systemic lupus erythematosus . J Exp Med 1990; 172: 653–656 .

Trowbridge IS , Thomas ML . CD45: an emerging role as a protein tyrosine phosphatase required for lymphocyte activation and development . Annu Rev Immunol 1994; 12: 85–116 .

Pingel JT , Thomas ML . Evidence that the leucocyte-commun antigen is required for antigen-driven T lymphocyte proliferation . Cell 1989; 58: 1055–1065 .

Thomas ML , Lefrancois L . Differential expression of leucocytecommon antigen family . Immunol Today 1988; 9: 320–326 .

Sato T , Furakawa K , Autero M et al. Structural study of the sugar chains of human leukocyte common antigen CD45 . Biochemistry 1993; 32: 12694–12704 .

Hathcock KS , Hirano H , Murakami S , Hodes RJ . CD45 expression by B cells. Expression of different CD45 isoforms by subpopulations of activated B cells . J Immunol 1992; 149: 2286–2294 .

10.

Caldwell CW , Patterson WP , Toalson BD , Yesus YW . Surface and cytoplasmic expression of CD45 antigen isoforms in normal and malignant myeloid cell differentiation . Am J Clin Pathol 1991; 95: 170–180 .

11.

Tan EM , Cohen AS , Fries JF et al. The 1982 revised criteria for the classification of systemic lupus erythematosus . Arthritis Rheum 1982; 25: 1271–1277 .

12.

Bombardier C , Gladman DD , Urowitz MB et al, and the committee on Prognosis Studies in SLE. Derivation of SLEDAI: a disease activity index for lupus patients . Arthritis Rheum 1982; 35: 630–640 .

13.

Hoffman RA , Recktenwald DJ , Vogt R . Cell-associated receptor quantitation. In. Bauer KD , Duque RE , Shankey TV (eds). Clinical Flow Cytometry: Principles and Applications. Williams & Wilkins: Baltimore, MD , 1993, pp 469–477.

14.

Bordron A , Dueymes M , Levy Y et al. Antiendothelial cell antibody binding induces endothelial cell apoptosis . J Clin Invest 1998; 101: 2029–2035 .

15.

Andrée HAM , Reutelingsperger CPM , Haupman R et al. Binding of vascular anticoagulant a to plasma phospholipid bilayers . J Biol Chem 1990; 265: 4923–4929 .

16.

Nicoletti I , Migliorati G , Pagliacci MC et al. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide and flow cytometry . J Immunol Meth 1991; 139: 271–279 .

17.

Czyzyk J , Fernsten P , Shaw M , Winfield JB . Cell-type specificity of anti-CD45 autoantibodies in systemic lupus erythematosus . Arthritis Rheum 1996; 39: 592–599 .

18.

Mimota S , Winfield JB . Identification of three major target molecules of IgM antilymphocyte autoantibodies in systemic lupus erythematosus . J Immunol 1987; 139: 3644–3650 .

19.

Bourgignon LYW , Suchard SJ , Nagpal MD , Clenny AT Jr . A T-lymphoma transmembrane glycoprotein (gp180) is linked to the cytoskeletal protein . J Cell Biol 1985; 101: 477–487 .

20.

Fernsten P , Shaw M , Hocker S et al. Expression of the sialosyl-Tn epitope on CD45 derived from activated peripheral blood T cells . Immunol Invest 1998; 27: 323–338 .

21.

Alsinet E , Inglés-Esteve J , Vilella R et al. Differential effects of anti-CD45 monoclonal antibody on human B cell proliferation: a monoclonal antibody recognizing a neuraminidase-sensitive epitope of the T200 molecule enhances anti-immunologulin-induced proliferation . Eur J Immunol 1990; 20: 2801–2804 .

22.

Fernsten PD , Czyzyk JK , Mimura T , Winfield JB . Carbohydrate specificity of IgM autoantibodies to CD45 in systemic lupus erythematosus . Mol Biol Rep 1994; 20: 85–95 .

23.

Pulido R , Sánchez-Madrid F . Glycosylation of CD45: carbohydrate composition and its role in acquisition of CD45RO and CD45 RB T cell maturation-related antigen specificities during biosynthesis . Eur J Immunol 1990; 20: 2667–2671 .

24.

Ohta T , Kitamura K , Maizel AL , Takeda A . Alterations in CD45 glycosylation pattern accompanying different cell proliferation states . Biochem Biophys Res Commun 1994; 200: 1283–1289 .

25.

Lefrancois L , Puddington L , Machamer CE , Bevan MJ . Acquisition of cytotoxic T lymphocyte-specific carbohydrate differentiation antigens . J Exp Med 1998; 162: 1275–1293 .

26.

Bernabeu C , Carrera AC , De Landazuri MO , Sánchez-Madrid F . Interaction between the CD45 antigen and phytohemagglutinin inhibitory effect on the lectin-induced T cell proliferation by anti-CD45 monoclonal antibody . Eur J Immunol 1987; 17: 1461–1466 .

27.

Yamada A , Cohen PL , Winfield JB . Subset specificity of antilymphocyte antibodies in systemic lupus erythematosus. Preferential reactivity with cells bearing the T4 and autologous erythrocyte receptor phenotype . Arthritis Rheum 1985; 26: 262–270 .

28.

Litvin DA , Cohen PL , Winfield JB . Characterization of warm-reactive IgG anti-lymphocyte antibodies in systemic lypus erythematosus. Relative specificity for mitogen-activated T cells and their soluble products . J Immunol 1983; 130: 181–186 .

29.

Klaus SJ , Sidorenko SP , Clark EA . CD45 ligation induces programmed cell death in T and B lymphocytes . J Immunol 1996; 156: 2743–2753 .

30.

Wu MX , Ao Z , Hegen M et al. Requirement of Fas (CD95), CD45 and CD11a/CD18 in monocyte-dependent apoptosis of human T cells . J Immunol 1996; 157: 707–713 .

31.

Salmon M , Pilling D , Borthwick NJ et al. The progressive differentation of primed T cells is associated with an increasing susceptibility to apoptosis . Eur J Immunol 1994; 24: 892–899 .

32.

Akbar AN , Terry L , Timms A et al. Loss of CD45R and gain of UCHL1 reactivity is a feature of primed T cells . J Immunol 1988; 140: 2171–2178 .

33.

Perillo N , Pace JJ , Seilhamer JJ , Baum LG . Apoptosis of T-cells mediated by galectin 1 . Nature 1995; 378: 736–738 .

34.

Stamenkoviv I , Sgroi D , Aruffo A et al. The B lymphocyte adhesion molecule CD22 interacts with leukocyte common antigen CD45RO on T cells and a 2 – 6 sialyltransferase CD75 on B cells . Cell 1991; 66: 1133–1144 .