Peptides of the SM-D antigen and their use for diagnosis of syst

Details Peptides of the SM-D antigen and their use for diagnosis of syst Peptides of the SM-D antigen and their use for diagnosis of syst

1992-04-03

1995-04-18

Nucker, Christine M.

Chemistry: analytical and immunological testing

For preexisting immune complex or auto-immune disease

Antinuclear

436538, 435 794, 435975, 530325, 530326, 530812, G01N 33564, C07K 1518, C07K 1708

Patent

active

054078334

DESCRIPTION:

BRIEF SUMMARY
The present invention concerns peptides capable of being recognized by antibodies present in biological fluids, in particular sera of patients or animals stricken with systemic lupus erythematosus (DLE).
The invention also concerns applications of these peptides, and compositions which contain them, to the in vitro diagnosis of systemic lupus erythematosus in humans, as well as their use in the composition of diagnostic kits.
The invention further concerns applications of these peptides in the production of immunogenic compositions and vaccination compositions against this disease.
Finally, the invention concerns antibodies which can be induced in vivo using these peptides, whether immunogenic or made immunogenic, and the application of these antibodies and the compositions containing them to in vitro diagnosis in humans stricken with DLE, as well as to the production of drugs used to fight this disease.
The presence of autoantibodies directed against cellular components is the general feature of auto-immune diseases such as systemic lupus erythematosus (DLE), scleroderma (Scl), polymyositis, and connective tissue disease (Morrow and Isenberg, 1987; Tan et al., 1988). Among the numerous kinds of autoantibodies identified for these disease, those which react with the Sm antigen constitute a very useful marker, since they are present in 20 to 30% of the subjects stricken with DLE, but are very rarely present in subjects suffering from other systemic autoimmune diseases of the connective tissues.
The Sm antigen is linked to a specific class of ribonucleoproteins, termed RNP's. These ribonucleoproteins contain five species of RNA rich in uridine, termed U1, U2, U4, US, and U6 (Lerner and Steitz, 1979; Brunel and al., 1985), in each of which seven proteins have been identified. Because of their electrophoretic mobility on polyacrylamide gels, they have been named band B' (29 Kd), band B (38 Kd), band D (16 Kd), band D' (15.5 Kd), band E (12 Kd), band F (11 Kd), and band G (9 Kd). In addition to these proteins, which make up the common nucleus, the U1-RNA particle contains three single polypeptides, corresponding, respectively, to 70 Kd, 34 Kd (A), and 22 Kd (C). The U2-RNP species contains two single polypeptides named A' (33 Kd) and B" (28.5 Kd). The anti-RNP antibodies, or anti UI-RNP, precipitate only the constituents of the U1 particle, while the anti-Sm antibodies react with the U1, U2, U4, US, and U6 particles.
Furthermore, immunoblotting studies have demonstrated that the anti-U1-RNP antibodies react with the A, C, and 70 Kd polypeptides, while the anti-Sm antibodies react with the B', B, and D polypeptides; the E, F, and G bands are also sometimes recognized by the anti-Sm antibodies (Pettersson et al., 1984; Reichlin and Harley, 1987; Hoch, 1989; Combe et al., 1989).
Sequencing of several polypeptides of the ribonucleoproteins was obtained using recombinant DNA techniques (Theissen et al., 1986; Habets et al., 1987; Stanford et al., 1987; Sillekens et al., 1987, 1989; Yamamoto et al., 1988; Rokeach et al., 1989). European Patent Application No. 295 719 describes the cloning of a DNA coding for the Sm-D antigen (Rokeach et al., 1988). This gene codes for a polypeptide of 119 amino acids containing several basic regions; the authors believe that a glycine-arginine unit repeated nine times and localized at the C-terminal end constitutes the antigenic determinant of the Sm-D antigen. This sequence, deduced from the isolated DNA, shows little similarity with the other, aforementioned sequenced polypeptides.
European Patent Application No. 295 719 describes, moreover, a method for detecting DLE which uses the cloned Sm-D antigen.
Work carried out by Applicant on the sequence of the Sm-D polypeptide containing 119 amino acids made it possible to conclude that certain peptide sequences selected from said Sm-D polypeptide are especially advantageous for the detection of DLE. Applicant observed that certain peptides deduced from the Sm-D polypeptide are recognized in a completely specific manner by antibodies pr

REFERENCES:
patent: 4629783 (1986-12-01), Cosand
patent: 4784942 (1988-11-01), Harley
Rokeach, L. A., et al., Proc. Natl. Acad. Sci., USA, 85(13):48321-4836, 1988.
Hubets, W. J., et al., Proc. Natl. Acad. Sci., USA, 86:4674-4678, 1989.

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