Apoptosis and expression of soluble Fas mRNA in systemic lupus erythematosus

Abstract

Our objective was to measure the relative proportions of messenger RNAs coding for soluble and membrane-bound Fas in peripheral blood mononuclear cells (PBMCs) and to quantify lymphocyte apoptosis in vitro in systemic lupus erythematosus (SLE).

A RT-PCR (soluble/membrane Fas mRNA) ratio was calculated in 16 SLE, 11 RA and 16 healthy subjects' PBMCs. Apoptosis was quantitated in vitro using nick-end labeling and flowcytometry analysis immediately and after overnight T cell activation.

The mean soluble/membrane Fas ratio was 1.219–0.424 in the SLE group, significantly higher than in healthy subjects, 0.516–0.180 (P = 0.0001). There was no significant difference between inactive and active SLE groups whereas Fas ratio was higher in SLE patients with nephritis (1.460–0.365) compared to those without nephritis (1.031–0.380) (P = 0.039). Mean soluble/membrane Fas ratio in RA patients (0.975–0.37) was higher than in healthy subjects (P = 0.0003) and lower than lupus nephritis patients (P = 0.015). A significantly higher proportion of mononuclear cells engaged apoptosis after T cell activation in active lupus patients, compared to control subjects.

In comparison with healthy subjects, Fas alternative splicing was skewed toward the soluble form of Fas in SLE and RA. Apoptosis after T cell activation in vitro was increased in active SLE patients.

Keywords

CD 95 lupus T lymphocyte nephritis

Get full access to this article

View all access options for this article.

References

Fraser A , Evan G . A license to kill . Cell 1996; 85: 781–784 .

Cheng J et al. Protection from Fas-mediated apoptosis by a soluble form of the Fas molecule . Science 1994; 263: 1759–1762 .

Chomczynski P , Sacchi N . Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction . Ann Biochem 1987; 162: 156–159 .

Itoh N et al. The polypeptide encoded by the cDNA for human cell surface antigen Fas can mediate apoptosis . Cell 1991; 66: 233–243 .

Pannetier C , Even J , Kourilsky P . T-cell repertoire diversity and clonal expansions in normal and clinical samples . Immunol Today 1995; 16: 176–181 .

Gonzalez EN , Rothfield NF . Immunoglobulin class and pattern of nuclear fluorescence in systemic lupus erythematosus . N Engl J Med 1966; 274: 1333–1338 .

Aarden CA , de Groot ER , Fedkamp TEW . Crithidiae luciliae, a simple substrate for the determination of anti ds-DNA with immunofluorescence technique . Ann NY Acad Sci 1975; 254: 505–515 .

Schur PH , Deangelis D , Jackson JM . Immunological detection of nucleic acids and antibodies to nucleic acids and nuclear antigens by counter-immunoelectrophoresis . Clin Exp Immunol 1974; 17: 209–218 .

Piqueras B , Autran B , Debré P , Gorochov G . Detection of apoptosis at the single cell level by direct incorporation of fluorescein-dUTP in DNA strand breaks . Biotechniques 1996; 20: 634–640 .

10.

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

11.

Bombardier C et al. Derivation of the SLEDAI. A disease activity index for lupus patients . Arthritis Rheum 1992; 35: 630–639 .

12.

Arase H , Arase N , Saito K . Fas-mediated cytotoxicity by freshly isolated natural killer cells . J Exp Med 1995; 181: 1235–1238 .

13.

Dhein J et al. Autocrine T-cell suicide by APO-1/(Fas/CD95) . Nature 1995; 373: 438–441 .

14.

Ju ST et al. Fas(CD95)/FasL interactions required for programmed cell death after T-cell activation . Nature 1995; 373: 444–448 .

15.

Nagata S , Suda T . Fas and Fas ligand: lpr and gld mutations . Immunol Today 1995; 16: 39–43 .

16.

Ito RM et al. Rheumatic diseases in an MRL strain of mice with a deficit in the functional Fas ligand . Arthritis Rheum 1997; 40: 1054–1063 .

17.

Fisher GH et al. Dominant interfering Fas gene mutations impair apoptosis in a human autoimmune lymphoproliferative syndrome . Cell 1995; 81: 935–946 .

18.

Rieux-Laucat F et al. Mutations in Fas associated with human lymphoproliferative syndrome and autoimmunity . Science 1995; 268: 1347–1349 .

19.

Le Deist F et al. Clinical, immunological, and pathological consequences of Fas-deficient conditions . Lancet 1996; 348: 719–723 .

20.

Drappa J et al. Fas gene mutations in the Canale-Smith syndrome: an inherited lymphoproliferative disorder associated with autoimmunity . N Engl J Med 1996; 335: 1643–1649 .

21.

Sneller MC et al. Clinical, immunologic, and genetic features of an autoimmune lymphoproliferative syndrome associated with abnormal lymphocyte apoptosis . Blood 1997; 89: 1341–1348 .

22.

Mysler E et al. The apoptosis-1/Fas protein in human systemic lupus erythematosus . J Clin Invest 1994; 93: 1029–1034 .

23.

McNally J et al. Fas ligand expression and function in systemic lupus erythematosus . J Immunol 1997; 159: 4628–4636 .

24.

Wu J et al. Fas ligand mutation in a patient with systemic lupus erythematosus and lymphoproliferative disease . J Clin Invest 1996; 98: 1107–1113 .

25.

Goel N et al. Lack of correlation between serum soluble Fas/APO-1 levels and autoimmune disease . Arthritis Rheum 1995; 38: 1738–1743 .

26.

Knipping E et al. Levels of soluble Fas/APO-1/CD95 in systemic lupus erythematosus and juvenile rheumatoid arthritis . Arthritis Rheum 1995; 38: 1735–1737 .

27.

Jodo S et al. Serum levels of soluble Fas/APO-1 (CD95) and its molecular structure in patients with systemic lupus erythematosus (SLE) and other autoimmune diseases . Clin Exp Immunol 1997; 107: 89–95 .

28.

Nozawa K et al. Soluble Fas (APO-1, CD95) and soluble Fas ligand in rheumatic diseases . Arthritis Rheum 1997; 40: 1126–1129 .

29.

Wolf RE , Brelsford WG . Soluble interleukin-2 receptors in systemic lupus erythematosus . Arthritis Rheum 1988; 31: 729–735 .

30.

Savill J , Henson P , Haslett C . Phagocyte recognition of cells undergoing apoptosis . Immunol Today 1993; 14: 131–136 .

31.

Emlen W , Niebur J , Kadera R . Accelerated in vitro apoptosis of lymphocytes from patients with systemic lupus erythematosus . J Immunol 1994; 152: 3685–3692 .

32.

Lorenz HM et al. In vitro apoptosis and expression of apoptosis-related molecules in lymphocytes from patients with systemic lupus erythematosus and other autoimmune diseases . Arthritis Rheum 1997; 40: 306–317 .

33.

Van Houten N , Budd RC . Accelerated programmed cell death of MRL-lpr/lpr T lymphocytes . J Immunol 1992; 149: 2513–2517 .

34.

Yoshino T et al. Inverse expression of bcl-2 protein and Fas antigen in lymphoblasts in peripheral lymph nodes and activated peripheral blood T and B lymphocytes . Blood 1994; 83: 1856–1861 .

35.

Sato T , Irie S , Kitada S , Reed JC . FAP-1: a protein tyro-sine phosphatase that associates with Fas . Science 1995; 268: 411–415 .

36.

Golstein P . Controlling cell death . Science 1997; 275: 1081–1082 .

37.

Georgescu L , Vakkalanka RK , Elkon KB , Crow MK . Interleukin-10 promotes activation-induced cell death of SLE lymphocytes mediated by Fas ligand . J Clin Invest 1997; 100: 2622–2633 .

38.

Rathmell JC et al. CD95 (Fas)-dependant elimination of self-reactive B cells upon interaction with CD4 T cells . Nature 1995; 376: 181–184 .

39.

Richardson BC et al. Monocyte apoptosis in patients with active lupus . Arthritis Rheum 1996; 39: 1432–1434 .