FR2672616A1 - Nucleotide sequences encoding variable regions of alpha chains of the receptors of human T lymphocytes, corresponding peptide segments and the diagnostic and therapeutic applications - Google Patents

Abstract

The present invention relates to new nucleotide sequences encoding the variable regions of alpha chains of the receptors of the human T lymphocytes, the corresponding peptide segments and the diagnostic and therapeutic applications.

Description

 The present invention relates to novel nucleotide sequences encoding variable regions of T cell receptor chains, corresponding peptide segments and diagnostic and therapeutic applications.

 It is known that receptors recognizing antigens on the surface of mature T lymphocytes (hereinafter referred to as T-cell receptors) have a structure that is somewhat analogous to those of immunoglobulins. Thus, they include heterodimeric structures with glycoprotein chains and glycoprotein chains (see Meuer et al (1), Moingeon et al.

(2), Brenner et al. (3), Bank et al. (4)).

The repertoire of T cell receptors must be able to cope with the immense diversity of antigenic determinants. This is obtained by genetic recombination of the different discontinuous segments of the genes which encode the different structural regions of the T cell receptors. Thus, the genes comprise V segments (variable segments), possibly D segments (diversity segments), segments J (joining segments) and segments
C (constant segments). During T-cell differentiation, specific genes are created by recombination of the V, D and J segments for the locusp and f and V and J segments for the A and B loci.
These specific combinations as well as the pairing of the two chains create combinatorial diversity.

This diversity is strongly amplified by two additional mechanisms, namely the imprecise union of the V-D-J or V-J segments and the addition of nucleotides corresponding to the N region (Davis et al.

 (5)).

 A number of V gene segments are already known. These segments have been grouped into sub-families according to the similarity of the sequences. By definition, segments with more than 75% similarity in the nucleotide sequence were considered members of the same subfamily (-Crews et al., 6)). The known gene segments V g have thus been classified in. 22 subfamilies including 14 with one limb (see Concannon et al (7), Kimura et al (8), Wilson et al (9)).

 In addition, about 60 J gene segments have been described (9).

 Furthermore, WO 90/06758 has recently described monoclonal antibodies directed against specific segments of the variable portions of the T cell receptors, in particular C / I and R caines.

These monoclonal antibodies are useful not only as diagnostic tools but also as therapeutic tools, for example with respect to rheumatoid arthritis.

 The use of synthetic peptides corresponding to g or ss variable regions in the treatment of autoimmune diseases (23 and 24) has also been described.

 It is also known that there are variations from one individual to another in the expression of the different variable segments of the human T receptor (27 and 28).

The present invention aims to enrich the repertoire of gene segments coding for variable regions of chains of T cell receptors by providing on the one hand new V & alpha gene segments; belonging to new sub-families or belonging to sub-families of which we already know at least one member and secondly, new genetic segments J
The present invention thus relates to nucleotide sequences coding for variable regions of α; lymphocyte receptors
Human T, corresponding to cDNAs comprising nucleotide sequences selected from any one of V <segments corresponding to one of SEQ ID N 1 to 11, and
b - segments J corresponding to one of the sequences SEQ ID N 12 to 20, and the sequences which differ from it by one or more nucleotides.

The present invention more particularly relates to:
sequences coding for variable regions of the &alpha; lymphocyte receptors
Human T, corresponding to cDNAs comprising nucleotide sequences chosen from any one of the segments V b (corresponding to one of the sequences SEQ ID Nos. 1 to 10, and the sequences which differ therefrom by one or more nucleotides.

sequences coding for variable regions of the &alpha; lymphocyte receptors
Human T, corresponding to cDNAs comprising nucleotide sequences selected from any one of J <segments corresponding to one of SEQ ID N 12 to 20, and sequences differing therefrom by one or more nucleotides.

 By the expression "and the sequences which differ therefrom by one or more nucleotides", we include alleles which have up to 8 nucleotides of difference, but most often 1 or 2 nucleotides of difference or which may differ by the deletion or the addition of one or two codons.

The present invention also more particularly relates to
nucleotide sequences coding for variable regions of the &alpha; human T cell receptors corresponding to cDNAs corresponding to all or part of the nucleotide sequences selected from any one of the V & alpha segments; corresponding to one of the sequences SEQ ID N '1 to 5, and the sequences which differ therefrom by one or two nucleotides,
nucleotide sequences coding for variable regions of human T lymphocyte receptor 4 chains corresponding to all or part of the nucleotide sequences chosen from any one of the segments corresponding to one of the sequences
1 to 467 of SEQ ID N 6
1 to 77 of SEQ ID N 7
1 to 151 of SEQ ID NO: 8
291 to 386 of SEQ ID N 9
1 to 260 of SEQ ID No. 10 and the sequences which differ therefrom by one or two nucleotides,
nucleotide sequences coding for variable regions of Q chains (human T cell receptors corresponding to cDNAs corresponding to all or part of the nucleotide sequence corresponding to SEQ ID No. 11 and which comprise nucleotide 108,
nucleotide sequences coding for variable regions of human T cell receptor chains corresponding to cDNAs corresponding to all or part of the nucleotide sequences chosen from any of the JA segments corresponding to one of the sequences SEQ ID N 12 at 20 and the sequences differing therefrom by one or two nucleotides.

 The expression "nucleotide sequences corresponding to cDNAs corresponding to all or part of the nucleotide sequences" denotes both the complete sequences and fragments of these sequences, including short fragments (oligonucleotides) which find applications as probes. (generally having at least 10 nucleotides) or as a primer (generally having at least 15 nucleotides). In general, the invention encompasses all the novel oligonucleotides which are fragments of the V sequences and according to the invention.

Sequences that differ in one or two nucleotides, they correspond to variations observed experimentally
CDNA.

 The present invention also relates to the peptides encoded by nucleotide sequences according to the invention as well as alleles and derivatives thereof which have the same function.

 The subject of the present invention is also the peptides constituted or comprising a peptide sequence encoded by all or part of the sequence 108 to 364 of SEQ ID No. 11.

 In general, the present invention encompasses peptides constituted by or comprising a peptide sequence encoded by the nucleotide sequences according to the invention as well as the fragments of these peptides. It also encompasses peptides which differ from one or more amino acids and which have the same function. These peptides may correspond to modifications such as those known with muteins or to allelic variations. In particular, it has been shown that certain gene segments coding for variable T-chain regions in humans are subject to a phenomenon of genetic polymorphism called allelic variation (25). The present invention encompasses peptides derived from this phenomenon.

The nucleotide sequences according to the invention were obtained according to the following steps
- isolation of peripheral lymphocytes RNA from an individual
obtaining the complementary DNA using reverse transcriptase and a primer A specific for the C <region (SEQ ID No. 21)
- gene amplification (by Anchored
Polymerase Chain Reaction or A-PCR) using a DNA polymerase, a poly C primer (SEQ ID No. 22) and a B primer specific for the Co region ((SEQ ID No. 23)).
new amplification by A-PCR using DNA polymerase and a primer C specific for the C & alpha region (SEQ ID No. 24)
- insertion in a plasmid vector
transformation of a bacterial host with the recombinant vector;
screening of the recombinant bacterial colonies with a labeled oligonucleotide D (SEQ ID No. 25)
extraction of the plasmids from the positive colonies,
and sequencing the DNA fragments containing the C region.

 The present invention may be reproduced in particular by bispecific gene amplification (polymerase chain reaction or PCR) starting from the peripheral lymphocytes expressing the mRNAs including the variable or junctional segments corresponding to the sequences ID Ne 1 to 20 of the invention or alternatively by applying this technique from PCR to genomic DNA from any somatic cell of a randomly selected individual. The invention can be reproduced by preparing the above gene sequences by chemical synthesis of oligonucleotides.

 The peptides according to the invention can be obtained by conventional peptide synthesis. They can also be obtained by applying known genetic engineering techniques including the insertion of a DNA sequence coding for a peptide according to the invention into an expression vector such as a plasmid and the transformation of cells with this invention. expression vector.

 The subject of the present invention is therefore also plasmids and expression vectors comprising a DNA sequence coding for a peptide according to the invention as well as hosts transformed with this vector.

 The subject of the present invention is also antibodies, and in particular monoclonal antibodies, directed against an antigenic determinant belonging to or comprising a peptide according to the invention.

 Monoclonal antibodies can be obtained by any of the techniques that allow the production of antibody molecules from cell line culture. These techniques include the different techniques using hybridomas.

 The production of antibodies can be obtained in animals by immunizing the animals by injection of the peptides or fragments according to the invention, whether natural, recombinant or synthetic, optionally after coupling with an immunogenic agent such as tetanus toxoid, or by injection of human T lymphocytes expressing the corresponding sequences on their surface, including recombinant cells transfected with the corresponding coding sequences.

 The subject of the present invention is also hybridomas producing monoclonal antibodies directed against the polypeptides according to the invention.

The present invention also encompasses the monoclonal antibody fragments and derivatives according to the invention which are reactive with defined variable regions of the T cell receptors. These fragments are notably the F (ab ') 2 fragments which can be obtained by cleavage. enzymatic antibody molecules with pepsin, fragments
Fab 'which can be obtained by reduction of disulfide bridges of F (ab') 2 fragments and Fab fragments which can be obtained by enzymatic cleavage of antibody molecules with papain in the presence of a reducing agent. These fragments can also be obtained by genetic engineering.

 The monoclonal antibody derivatives are, for example, antibodies or fragments of these antibodies to which markers such as a radioisotope are linked. The monoclonal antibody derivatives are also antibodies or fragments of these antibodies to which therapeutically active molecules, in particular cytotoxic compounds, are bound.

 The products of the invention find several types of application in the field of diagnosis and in the field of therapy.

1 - Applications in the field of diagnosis
The oligonucleotides contained in the nucleotide sequences according to the invention can be used to constitute detection probes (generally at least 10 nucleotides) -capable of hybridizing to a variable region of the chain <or primers for amplification of DNA (generally comprising at least 15 nucleotides and preferably at least 17 nucleotides) capable of binding to a sequence to be amplified.

 The oligonucleotides thus find an application in the diagnosis of immune disorders by detecting the presence of nucleic acid sequences homologous to a gene encoding variable regions of T-cell receptor waves in the mRNA of a sample of a patient.

Different methods can be used to link T cell gene expression with disease. These methods include
a - the production and analysis of cDNA expression libraries obtained from disease-related T cells to determine the frequency of dominant genes
b - Southern blot analysis of genomic DNA samples to determine if there are genetic polymorphisms or rearrangements of genes encoding T cell receptors
c - analysis of samples by obtaining cDNA, PCR amplification and hybridization with labeled probes
d - in situ hybridization of T cells without prior culture of T cells.

 The primers find application in PCR reactions in a method such as that defined in c above.

 The monoclonal antibodies, the fragments or the derivatives of these antibodies according to the invention, in particular the anti-Vg antibodies, can be used to study T-type immune responses, for example in the field of autoimmune diseases of oncology, allergy, transplantation and infectious diseases. In particular, the repertoire of different variable segments i of the T-receptor can be studied, whether they are blood T-cells or tissues. In general, the techniques used can be in vitro or in vivo methods.

In in vitro methods, the samples used may be samples of body fluids or tissue samples. The techniques used may include flow cytofluorimetry to analyze lymphocytes
Blood T or immunoperoxidase markings on anatomopathological section to study tissue infiltrating lymphocytes.

 In in vivo methods, antibodies, fragments or derivatives thereof are administered by the usual routes, for example by the intravenous route, and the immunospecific binding is detected. This can be achieved for example in the case where a radioisotope labeled antibody is used.

2 - Applications in the therapeutic field
The oligonucleotides contained in the nucleotide sequences according to the invention can be used in therapy as antisense oligonucleotides. It is known that it is possible in vitro to inhibit the expression of a transcript gene in human lymphocytes by incubating these lymphocytes with an ante oligonucleotide in specific to the gene in question (26). These antisense oligonucleotides generally comprise at least 10 and, preferably, at least 16 nucleotides. These antisense oligonucleotides may be in particular the inverted and complemented sequences corresponding to the nucleotides upstream of the translation initiation site (ATG).

The advantage of an in vitro use of antisense oligonucleotides specific to a gene segment Zou
It is necessary to abolish (or greatly reduce) the expression of a receptor T comprising this segment V or J and thus to obtain a clonal deletion phenomenon at the level of the specific reactivity of the T lymphocytes.

The ante oligonucleotides can not only be used in vitro on human T lymphocytes which are then reinjected, but also in vivo by local or systemic injection, preferably after modification to increase the stability in vivo and the penetration inside the lymphocytes. T of these oligonucleotides.

The monoclonal antibodies according to the invention, in particular the anti V <antibodies, can be used to modulate the immune system. Thus the antibodies can be administered to block the interaction of effector T cells with their specific antigen. Anti-T-receptor antibodies bound for example to a cytotoxic molecule or a radioisotope can also be administered so as to obtain a clonal deletion by specific binding on a chain & a cell receptor.
T. The monoclonal antibodies according to the invention may be used therapeutically at low mitogenic concentrations so as to specifically activate certain subsets of T cells or may be used at much higher concentrations to bind to the relevant receptors and thus to mark these subsets for their elimination by the reticuloendothelial system. An important criterion for the treatment of a disease is the ability to modulate T-cell subsets related to a disease. The exact nature of this therapeutic modulation, namely blocking or suppressing a particular subset of T cells or, on the contrary, stimulating and activating a particular subset, will depend on the disease in question and the specific subset of T cells concerned.

This type of treatment has an advantage over current treatments using antibodies such as treatment with anti-CD3 antibodies in patients having a kidney transplant and having a problem of rejection, since thanks to the invention it is There will be no modulation of the T cell population but only the subset of T cells expressing the particular cell receptor subfamily.
T.

 Moreover, the response of T cells is often oligoclonal, it is generally appropriate to use in therapy # cocktails of several antibodies.

In addition, the anti-antibodies can be used
V g to select in vitro T lymphocytes, for example by passing on a column containing beads carrying the antibody. This separation of certain T lymphocytes can be used for culturing these lymphocytes before reinjecting them to the patient.

 In addition, all or part of the peptide sequences according to the invention can be used therapeutically, that is to say the peptide sequences encoded by the nucleotide sequences according to the invention or the fragments of these sequences (generally comprising at least 8 to 10 amino acids). These sequences or fragments, administered to humans or animals, can act as decoys, that is to say they will bind to the epitope carried by the harmful antigen and prevent the reaction of normal T cells with the antigen, thus preventing the development of an aggressive disease against self determinants. They can also be used as immunogens in the manufacture of vaccines (possibly after coupling to protein carriers).

The following will be described in more detail below, the obtaining of the nucleotide sequences according to the invention, with reference to FIGS. annexed on which
- Figs. 1A to E give in alignment both a known sequence V i and a partial sequence of an extension according to the invention for the respective sequences SEQ ID N 6 to 10, denoted IGRa 08 to IGRa 12. In these figures, the nucleotide numbering begins at the initiation ATG codon (which is underlined). The dots indicate identical nucleotides. The sequences that are supposed to be leader sequences are highlighted.

FIG. 2 gives in alignment the new sequences J '(SEQ ID NO 12 to 20) denoted IGR & O1 to
IGRJa 09. In these sequences the recombination signals of the germ line are underlined. The amino acids corresponding to the highly conserved codons are marked above the sequences. The codons corresponding to a substitution at a position of a conserved amino acid are doubly underlined.

 FIG. 3 gives Southern blot analysis of genomic DNA treated with a restriction enzyme using probes specific for SEQ ID NO: 1 to 5. The restriction enzymes used are EcoRI (R column), Hind III (H column) and Bam III (column B). In this Figure, the sorting groups mark the position of the DNA fragments hybridizing specifically with C.

I - Obtaining the cDNA and PCR amplification
RNA sources were peripheral lymphocytes of an individual.

 Total RNA was prepared according to the guanidinium isothiocyanate and cesium chloride method (Chirgwin (10)) or the one-step method by extraction with guanidinium isothiocyanate, phenol and chloroform (Chomczynski (11)).

The first strand of cDNA was synthesized in a final volume of 50 microliters at a temperature of 42 ° C for 1 hour using 5 micrograms of total RNA, reverse transcriptase and a primer A specific for the C 4 region consisting of the sequence 5'-GTTGCTCCAGGCCACAGCACTG (SEQ ID N 21) This material was then purified by phenol / chloroform extraction and ammonium acetate precipitation After selection of a 0.45 / 1 kb gel fraction of agarose, the addition of a dG end to the RNA / cDNA hetero duplex in addition buffer was performed.
CoCl2 with 14 units of deoxynucleotidyl transferase (Tdt) terminal for 30 minutes at 37 ° C. The reaction was stopped by holding at 70 ° C. for 10 minutes. NaOH
IN (13% volume) was added and the sample incubated at 50 ° C for 1 hour to hydrolyze the RNA and then neutralized with 2M Tris HCl pH 8 and 1N HCl. After extraction with a phenol / chloroform mixture, the first strand of G-end cDNA was precipitated with ethanol and amplified using PCR (Polymerase Chain
Reaction described by Saiki et al. (12)) in a final volume of 100 microliters containing 50 mM KCl, 10 mM Tris-Cl pH 8.3, 1.5 mM MgCl 2, 0.1% (w / v) gelatin, 200 micromoles dNTP, 2.5 units of Taq polymerase and 100 picomoles of two primers. The two primers used are, on the one hand, a poly-C primer (5'-GCATGCGCGCGGCCGCGGAGG-14C) (SEQ
ID N 22) described by Loh et al. (13) and a CO-specific primer B ((5'-GTCCATAGACCTC
ATGTCCAGCACAG) (SEQ ID No. 23).

 25 amplification cycles are carried out followed by a period of 15 minutes of final elongation at 72 ° C.

Each cycle includes a denaturation step at 92
C for 1 minute, a hybridization step at 55 ° C for 2 minutes and an elongation period at 72 ° C. for 4 minutes The amplified products are then precipitated with ethanol, resuspended in acetate 30mM sodium pH5, 50mM NaCl, 1mM ZnCl2, 5% v / v glycerol, and 1/10 of this material is size-purified on a 1% low melting agarose gel.

 A second amplification phase is then carried out directly on approximately 10% of the band containing the agarose according to the same conditions as above, except that C-region-specific primer C <is used as the 5'-ATACACATCAGAATTCTTACTTTG primer. (SEQ ID N 24). The reaction mixture is then precipitated with ethanol and resuspended in 60 l H2O.

II - Clonaae and sequence of cDNAs
1/3 of the product of the second amplification is digested with Sac II, separated on 1% agarose gel and purified by absorption on glass beads. The material is inserted into the vector
Bluescript SK (Stratagene, La Jolla, USA) and the recombinants obtained are used to transform E. coli strains XL1-blue (Stratagene). After deposition in the presence of λ-galactosidase and IPTG, a test is carried out on the white colonies using a "dot blot" technique and as a probe, a third oligonucleotide specific for the C4 region (5I-GTCACTGG
ATTTAGAGTCT) (SEQ ID NO: 25) labeled at 32p. Plasmid DNA was extracted from the positive colonies and sequenced under both strands by the dideoxy chain termination method (Sanger et al (14)) with Sequenase. 2.0 (United States Biochemicals,
Cleveland, USA) according to the supplier's recommendations. Except for SEQ ID No. 5, all nucleotide sequences were determined on both strands using at least two distinct cDNA clones.

 The resulting sequences were compared to published sequences V i and J i using the method developed by Lipman and Pearson (15). The presumed start codons were identified by searching for the presence of the Kozak consensus sequence for translation initiation sites in eukaryotic cells (Kozak (16)). The presence of hydrophobic leader sequences on the N-terminal side was detected by analysis of the hydrophobicity according to the method described by Kyte (17 !.

III - Analvse By Southern blot
The DNA was extracted from the human erythroleukemic cell line K562 and digested with one of the following restriction enzymes: EcoRI,
Bam HI or Hind III. DNA (15 micrograms) was electrophoresed on 0.7% agarose and transferred to nylon membranes as described by
Triebel et al. (18). Hybridizations were performed at 65 ° C with 6 X SSC, 0.5% SDS, 5 X
Denhardt's and 100 micrograms of denatured salmon sperm DNA for 16 hours. The membranes were washed at 65 ° C with 2 X SSC, 0.2% SDS.

 V & alpha probes were used; specific probes obtained by amplification of V-J-C cDNA (> 500 bp) comprising V & alpha fragments; corresponding to sequences SEQ ID N 1 to 5 using as primer the primer poly-C and primer C. The probes were purified on agarose gel 1%. 32-labeled DNA probes were prepared from the agarose-purified fragments by the Feinberg method (19).

IV - Results
Using the A-PCR method, 308 cDNAs that hybridize with the C & alpha probe; were cloned, then sequenced. Of these, 172 cDNAs correspond to unique VJ-Co variable regions.

 Met J & alpha sequences; of the invention are listed in the sequence list respectively under SEQ ID Nos. 1 to 11 and SEQ ID Nos. 12 to 20. Sequence sequences SEQ ID No. 1 to 5 correspond to new subfamilies while sequences SEQ ID No. 6 to 11 correspond to V & alpha segment extensions; known.

1. V & alpha sequences; corresponding to. new subfamilies
Southern blot analyzes of endonuclease-digested germline DNA using VJC & alpha probes; comprising V & alpha fragments; corresponding to SEQ sequences
ID N 1 to 5 were performed under "low stringency" hybridization conditions to identify the number of V gene segments belonging to each family and to characterize DNA restriction fragments carrying these V & alpha gene segments;
Representative results are reported on the
Figure 3

 These analyzes show that the subfamily corresponding to the sequence SEQ ID No. 3 comprises at least two gene segments while the other sequences (SEQ ID No. 1, N 2, N 4 and N 5) probably correspond to single members.

The sizes of the restriction fragments of the germinal DNA are as follows
SEQ ID N 1: Eco RI 4.7 kb, Hind III 3.7 kb
SEQ ID NO: 2 EcoRI 2.2 kb, Hind III 4.8 and 5.7 kb,
Bam HI 25 kb
SEQ ID N-3: Eco RI 4.6 and 7.5 kb, Hind III 4.2 and
6.4 kb, Bam HI 23 and 4.5 kb
SEQ ID N 44: Eco RI 7.6 kb, Hind III 18 kb, Bam HI 9
and 0.9 kb
SEQ ID NO: 5: Eco RI 5.9 and 4.8 kb, Hind III 6.6 kb,
Bam HI 6.5 kb.

2. Sequences corresponding to extensions of V & alpha sequences; known
SEQ ID No. 6 (IGR a 08): This sequence corresponds to an extension on the 5 'side of the sequence of the V & alpha clone; 1 AE1f (Klein et al (21)). The two aligned sequences are shown in Figure 1A.

SEQ ID No. 7 (IGR a 09): This sequence corresponds to an extension coding for the NH 2 terminal end of the V 4 2 AFIII sequence (Klein already cited). The two aligned sequences are shown in FIG. 1B.

The sequence ID No. 7 corresponds to a consensus sequence. In position 206 the existence of a T instead of a C was observed.

SEQ ID NO: 8 (IGR at 010): This sequence corresponds to an extension of the 5 'region of the V & alpha clone; HAP 35 (Yoshikai (22)). The two aligned sequences are shown in FIG. 1 C. The sequence ID No. 8 corresponds to a consensus sequence. In position 307 we observed the existence of a G instead of an A and in position 360 the existence of a T instead of a C.

SEQ ID N-9 (IGR a 11). This sequence corresponds to an extension on the 3 'side of the V & alpha sequence; 7 HAP 12 (Yoshikai already cited). Sequence alignment is shown in FIG. 1D. The sequence ID N 9 corresponds to a consensus sequence. Position 86 has been observed to have a C instead of a T.

SEQ ID N 10 (IGR at 12): This sequence comprises the complete coding region of a gene of the subfamily V &alpha; 22 which had previously been identified by the partial sequence 113 bp AC9 (Klein already cited). The two aligned sequences are shown in FIG.

1E.

SEQ ID N-11 (IGRa 13): This sequence corresponds in part to the clones HAVT 32 and HAVT 35 (belonging to the subfamily V i 16 (8) and which have been described as pseudogenes. of the addition of a nucleotide at position 108, SEQ ID No. 11 encodes an original variable region of a T cell receptor.

3. Sequences J i
It is shown in FIG. 2 all the new sequences J g. Of the 9 J segments, most of them have a highly conserved amino acid sequence of FGXGT segments as described by Yoshikai already cited. However, for segment IGRJa 07 the threonine residue is replaced by an isoleucine residue.

 In addition, instead of the phenylanine residue, a cysteine residue was found in IGRJa 02G.

REFERENCES 1. Meuer, S.C., et al., J. Exp. Med. 1983. 157: 705.

2. Moingeon, P., et al., Nature 1986a, 323: 638.

3. Brenner, M.B., et al., Nature 1986. 322: 145.

4. Bank, I., et al., Nature 1986. 322: 179.

5. Davis, M. M., et al., Nature 1988.334: 395.

6. Crews, S., et al., Cell 1981. 25:59.

7. Concannon, P., et al., Proc. Natl. Acad. Sci. USA
1986. 83: 6598.

8. Kimura, N., et al., Eur. J. Immunol. 1987.

 17: 375.

Wilson, R.K., et al., Immunological Reviews 1988c.

 101: 149.

10. Chirgwin, J.M., et al., Biochemistry 1979.

 18: 5294.

11. Chomczynski, P. et al., Anal. Biochem. 1987
162: 156.

12. Saiki, R.K., et al., Science 1988. 239: 487.

13. toh, E.Y., et al., Science 1989. 243: 217.

14. Sanger, F., et al., Proc. Natl. Acad. Sci. USA
1977, 74: 5463.

15. Lipman, D.J., et al., Science 1985. 227: 1435.

16. Kozak, M., Nucl. Acids Res. 1984. 12: 857.

17. Kyte, J., et al., R.F., J. Mol. Biol. 1982.

 157: 105.

18. Triebel, F., et al., J. Immun. 1988. 140: 300.

19. Feinberg, A. P., et al., Anal. Biochem. 1983.

 132: 6.

20. Mengle-Gaw, L., et al., The EMBO Journal, 1987.

 6: 2273.

21. Klein, MH, et al., Proc. Natl. Acad. Sci. USA
1987, 84: 6884.

-22. Yoshikai, Y., et al., J.-Exp. med. 1986. 164: 90 23. Naudenbark A., et al., Nature, 341, 541.

24. Janeway C., Nature, 341, 482.

25. Lin Y., J. Exp. Med., 171, 221.

26. Acha-Orbea H., EMBO Journal, 1990, 9, 12, 3815.

27. Kappler J., Science, 244, 811 28. Choi, Y., PNAS, 86, 8941.

LIST OF SEQUENCES
I - GENERAL INFORMATION
(1) APPLICANT: Company known as ROUSSEL UCLAF
(2) TITLE OF THE INVENTION
Nucleotide sequences coding for
variable regions of the &alpha; receptions
human T lymphocytes, segments
corresponding peptides and applied them
diagnostic and therapeutic

(3) NUMBER OF SEQUENCES: 25
II - INFORMATION FOR SEQ ID N 1
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 371 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: - IGR a 02
SEQUENCE V &alpha;
DESCRIPTION OF THE SEQUENCE
AGTCAACTTC TGGGAGCAGT CTCTGCAGAA TAAAA ATG AAA CAT AAG 47
Met Lys Lys His
1
CTG ACG ACC TTC TTG GTG AT TTG TGG CTT TAT TT TAT AGG GGG AAT 95
Leu Thr Thr Phe Leu Val Ile Leu Trp Leu Tyr Phe Tyr Arg Gly Asn 5 10 15 20
GGC AAA AAC CAA GTG GAG CAG AGT CCT CAG TCC CTG ATC ATC CTG GAG 143
Gly Lys Asn Gln Gln Val Glu Ser Gln Ser Pro Leu Ile Ile Glu Glu
25 30 35
GGA AAG AAC TGC ACT CTT CAA TGC AAT TAT ACA GTG AGC CCC TTC AGC 191
Gly Lys Asn Cys Thr Leuk Gln cys Asn Tyr Thr Ser Val Pro Phe Ser
40 45 50
AAC TTA AGG TGG TAT AAG CAA ACT GGG AGA GGT CCT GTT TCC CTG 239
Asn Leu Arg Trp Tyr Lys Asp Gln Gly Arg Arg Gly Pro Val Ser Leu
55 60 65
ACA ATC ATG ACT TTC AGT GAG AAC ACA AAG TCG AAC GGA AGA TAT ACA 287
Thr Met Thr Phe Ser Glu Asn Thr Lys Ser Asn Gly Arg Tyr Thr
70 75 80
GCA ACT CTG GAT GCA ACA GAC ACG CAA AGC TCT CTG CAC ATC ACA GCC 335
Ala Thr Leu Asp Ala Asp Thr Lys Gln Ser Ser Leu His Isle Thr Ala 85 90 95 100
TCC CAG CTC AGC GAT TCA GCC TCC TAC ATC TGT GTG 371
Ser Gln Ser Ser Asp Ser Ser Ala Ser Isle Cys Val
105,110
III - INFORMATION FOR SEQ ID N 2
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 400 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T cells CHARACTERISTICS
DNA NAME: IGR a 03
SEQUENCE V &alpha;
DESCRIPTION OF THE SEQUENCE
GACTCTAAGC CCAAGAGAGT TTCTTGAAGC AAAAAAAAAA 40
AAAACCCATT CAGGAAATAA TTCTTTGCTG ATAAGG ATG CTC CTT GAA CAT TTA 94
Met Leu Leu Glu His Leu
1 5
TTA ATA ATC TTG TGG ATG CAG CTC ACA TGG GTC AGT GGT CAA CAG CTG 142
Leu Ile Leu Trp Island Net Gln Leu Thr Trp Ser Val Glly Gln Gln Leu
10 15 20
AAT CAG ACT CCT CAA TCT ATG TTT ATC CAG GAA GGA GAAT GAT GTC TCC 190
Asn Gln Ser Gln Gln Ser Met Phe Gln Glu Gly Asp Glu Gl
25 30 35
ATG AAC TGC ACT TCT TCA AGC ATA TTT ACC AAC TGG CTA TGG TAC AAG 238
Met Asn Cys Ser Ser Ser Ser Phe Asn Thr Trp Leu Trp Tyr Lys
40 45 50
CAG GAC CCT GGG GAA GGT CCT GTC CTC TTC GCC ATA GTA TAT AAG GCT 286
Gln Asp Pro Gly Glu Gly Pro Val Leu Leu Ile Ala Leu Tyr Lys Ala
55 60 65 70
GGT GAA TTC ACC TCA AAT GGA AGA CTC ACT CCT CAG TT GGT ATA ACC 334
Gly Glu Leu Thr Ser Asn Gly Arg Leu Thr Ala Gln Phe Gly Thr Island
75 80 85
AGA AAG GAC AGC TTC CTG AAT. ATC TCA GCA CBT ATA CCT AGT GAT GTA 382
Arg Lys Asp Ser Phe Leu Asn Ser Ser Ala Ser Pro Ser Asp Asp Val
90 95 100
GGT ATC TAC TTC TGT GCT 400
Gly Island Tyr Phe Cys Ala
105
IV - INFORMATION FOR SEQ ID NO: 3
CHARACTERISTICS OF THE SEQUENCES
TYPE. nucleotide and its corresponding protein
ponding
LENGTH: 339 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF DNA: IGR a 04
SEQUENCE V
DESCRIPTION OF THE SEQUENCE
AGCTAAGGG ATG GAG ACT GTT CTG CAA GTA CTA CTC GGG ATA TTG GGG 48
Met Glu Thr Val Leu Gln Val Leu Leu Gly Ile Leu Gly
1 5 10
VAT 'GAA GCA GCC TGG GTC AGT AGC CAA GAA CTG GAG AGC AGT CCT CAG 96
Phe Gln Ala Ala Trp Ser Ser Gln Glu Leu Glu Ser Gln Pro Gln
15 20 25
TCC TTG ATC GTC CAA GAG GGA AAG AAT CTC ACC ATA AAC TGC ACG TCA 144
Ser Leu Val Gln Island Glu Gly Lys Asn Leu Thr Island Asn Cys Thr Ser 30 35 40 45
TCA AAG ACG TTA TAT GGC TTA TAC TGG TAT AAG CAA AAG TAT GGT GAA 192
Ser Lys Thr Leu Tyr Gly Leu Tyr Trp Tyr Lys Gln Lys Tyr Gly Glu
50 55 60
GGT CTT ATC TTG TTG ATG ATC CTA CAG AAA GGT GGG GAA GAG AAA AGT 240
Gly Leu Ile Phe Leu Met Met Gln Lys Gly Gly Glu Glu Lys Ser
65 70 75
CAT GAA AAG ATA ACT GCC AAC TTC GAT GAG AAA AAG CAG CAA AGT TCC 288
His Glu Lys Thr Island Ala Lys Leu Asp Glu Lily Lys Gln Gln Ser Ser
80 85 90
CTG CAT ATC ACA GCC TCC CAG CCC AGC CAT GCA GGC ATC TAC CTC TGT 336
Leu His Island Thr Ala Ser Gln Pro Ser His Ala Gly Ile Tyr Leu Cys
95 100 105
GGA 339
Gly 110
V - INFORMATION FOR SEQ ID N 4
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 335 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF DNA: IGR a 05
SEQUENCE V &alpha;
DESCRIPTION OF THE SEQUENCE
AGAAAAAAAA AATGAAGAAG CTACTAGCAA TGATCCTGTG GCTTCAACTA 50
GACCGGTTAA GTGGAGAGCT GAAAGTG GAA CAA AAC CCT CTG TTG 95
Glu Asn Gln Asn Pro Phe
1 5
CTG AGC ATG CAG GAG GGA AAA ATAC TAT ACC ATC TAC TGC AAT TAT TCA 143
Leu Ser Met Gln Glu Gly Lys Asn Tyr Thr Tire Tyr cys Asn Tyr Ser
10 15 20
ACC ACT TCA GAC AGA CTG TAT TGG TAC AGG CAG GAT CCT GGG AAA AGT 191
Thr Thr Ser Asp Asp Arg Tyr Tyr Trp Tyr Arg Asp Gln Gly Lys Ser
25 30 35
CTG GAA TCT CTC TT GTG TTG CTA TCA AAT GGA GTG GTG AAC CAG GAG 239
Leu Leu Ser Leu Phe Leu Leu Leu Ser Asn Gly Ala Val Lys Gln Glu
40 45 50
GGA CGA TTA ATG GCC TCA CTT GAT ACC AAA GCC CGT CTC AGC ACC GTG 287
Gly Arg Leu Met Ala Ser Leu Asp Thr Lys Ala Arg Leu Ser Thr Leu
55 60 65 70
CAC ATC ACA GCT GCC GTG CAT GAC CTC TCT GCC ACC TAC TTC TGT GCC 335
His Thr Island Ala Ala Val His Asp Al Ser Leu Thr Tyr Phe Cys Ala
75 80 85
VI - INFORMATION FOR SEQ ID N 5
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 361 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF DNA: IGR a 07
SEQUENCE V &alpha;
DESCRIPTION OF THE SEQUENCE
GAAGCTGACT GGATATTCTG GCAGGCCAAG ATG GAG ACT CTC CTG 46
Met Glu Thr Leu Leu 1 Es
AAA CTG CCT TCA GGC ACC TTC TTC TGG CAG TTC ACC TGG GTG GGA AGC 94
Lys Val Pro Ser Gly Thr Leu Leu Trp Gln Leutr Trp Val Gly Ser
10 15 20
CAA CAA CCA GTG CAG AGT CCT CAA GCC GTG ATC CTC CGA GAA GGG GAA 142
Gln Gln Pro Gln Gln Pro Val Gln Ala Val Glu Glu Gly Glu
25 30 35
GAT GCT GTC ACC AAC TGC AGT TCC TCC AGG GCT TTA TAT TCT GTA AGC 190
Asp Ala Val Thr Asn Cys Ser Ser Ser Lys Ala Leu Tyr Ser Val His
40 45 50
TGG TAC AGG CAG AAG CAT GGT GAA GCA CCC GTC TTG CTG ATG ATA TTA 238
Trp Tyr Arg Gln Lily His Gly Glu Ala Pro Val Phe Leu Met Ile Leu
55 60 65
CTG AAG GGT GGA GAA CAG ATG CGT GAA GAA AAA ATA TCT GCT TCA TT 286
Leu Lys Gly Gly Glu Gln Met Arg Arg Glu Lily Ser Ser Ala Ser Phe 70 75 80 85
AAT GAA AAA AAG CAG CAA AGC TCC CTG TAC CTT ACG GCC TCC CAG CTC 334
Asn Glu Lys Lys Gln Gln Ser Ser Leu Leu Leu Thr Ala Ser Gln Leu
90 95 100
AGT TAC TCA GGA TAC ACCT TTG TGC GGG 361
Ser Tyr Ser Gly Thr Tyr Phe Cys Gly
105,110
VII - INFORMATION FOR SEQ ID N 6
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 569 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
DNA NAME: IGR at 08
SEQUENCE V &alpha; 1
DESCRIPTION OF THE SEQUENCE
TCAGTTTCTT 10
CTTCCTGCAG CTGGTTGAGT TCTTTCCAGA CAAAGACAAG TGACAAGAAT TAGAGGTTTA 70
AAAAGCAACC AGATTCATCT CAGCAGCTTT TGTAGTTTTA AATAAGCAAG GAGTTTCTCC 130
AGCGAAACTT CCTCACACCT CTTGGTCTTG GTCTCTTCAG ACACTTTCCT TCCTGTTCTC 190
TGGAGATCTT GCAGAAAAGA GCCTGCAGTG TTTCCCTTGC TCAGCC ATG CTC CTG 245
Met Leu Leu
1
GAG CTT ATC CCA CTG CTG GGG ATA CAT TTT GTC CTG AGA ACT GCC AGA 293
Glu Leu Ile Pro Leu Leu Gly Ile His Phe Val Leu Arg Thr Ala Arg
5 10 15
GCC CAG TCA CTG ACC CAG CCT GAC ATC CAC ATC ACT GTC TCT GAA GGA 341
-Ala Gln Ser Thr Val Gln Pro Asp Her Island Isle Thr Val Ser Glu Gly
20 25 30 35
GCC TCA CTG GAG TTG AGM TGT AAC TAT TCC TAT GGG GCA ACA CCT TAT 389
Ala Ser Leu Glu Leu Arg Cys Asn Tyr Ser Tyr Gly Ala Thr Pro Tyr
40 45 50
CTC TTC TGG TAT GTC CAG TCC CCC GGC CAA GGC CTC CAG CGT CTC CTG 437
Leu Phe Trp Tyr Val Gln Ser Gly Gln Gly Leu Leu Leu Leu Leu
55 60 65
AAG TAC TTT TCA GGA GAC ACT CTG GTT CAA GGC ATT AAA GGC TTT GAG 485
Lys Tyr Phe Ser Gly Asp Thr Leu Val Gln Gly Ile Lys Gly Phe Glu
70 75 80
GCT GAA TTT AAG AGG AGT CAA TCT TCC TTC AAC CTG AGG AAA CCC TCT 533
Ala Glu Phe Lys Arg Ser Ser Gln Ser Phe Asn Leu Ser Arg Lys Pro Ser
85 90 95
GTG CAT TGG AGT GAT GCT GCT GAG TAC TTC TGT GCT 569
Val His Trp Asp Asp Ala Ala Glu Tyr Phe Cys Ala
100 105 110
VIII - INFORMATION FOR SEQ ID N 7
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 330 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
SEQUENCE CONSENSUS
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
DNA NAME: IGR a 09
SEQUENCE V &alpha; 2
DESCRIPTION OF THE SEQUENCE
AAA TGG TTC AGA GTT TTA CTA GTG ATC CTG TGG CTT AGC GTC AGC CGG 48
Ser Leu Leu Arg Leu Leu Leu Leu Leu Leu Leu
1 5 10 15
GTT TGG AGC CAA CAG AGG GAG GTC GAG AGC AGT ATCT GCA CCC GTG AGT 96
Val Trp Ser Gln Gln Lys Glu Glu Val Glu Asn Ser Gly Pro Leu Ser
20 25 30
GTT CCA GAG GGA GCC ATT GCC TCT CTC AAC TGC ACT TAC AGT GAC CGA 144
Val Pro Glu Gly Ala Ile Ala Ser Asn Leu Cys Thr Tyr Asp Asp Asp Arg
35 40 45
GGT TGG CAG TCC TTG TTG TGG TAC AGA CAAT TAT TCT GGG AAA AGC CCT 192
Gly Ser Gln Ser Phe Phe Trp Tyr Arg Gln Ser Ser Gly Lys Ser Pro
50 55 60
GAG TTG ATA ATG TCC ATA TAC TCC AAT GGT GAC AAA GAAT GAT GGA AGG 240
Glu Leu Ile Met Ser Tire Asn Asly Gly Asp Lys Glu Asp Gly Arg
65 70 75 80
TTT ACA GCA CAG CTC AAT AAA GCC AGC CAG TAT GTT TCT CTG CTC ATC 288
Phe Thr Ala Gln Leu Asn-Lys Ala Ser Gln Tyr Val Ser Leu Leu Ile
85 90 95
AGA GAC TCC AGC CCC AGT GAT TCA GCC ACC TAC CTC TGT GCC 330
Arg Asp Ser Gln Pro Ser Asp Ser Ala Thr Tyr Leu Cys Ala
100 105 110
IX - INFORMATION FOR SEQ ID N 8
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 400 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
SEQUENCE CONSENSUS
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
DNA NAME: IGR has 10
SEQUENCE V &alpha; 5
DESCRIPTION OF THE SEQUENCE
GCCAAACAGA ATGGCTTTTT GGCTGAGAAG GCTGGGTCTA CATTTCAGGC 50
CACATTTGGG GAGACGA ATG GAG TCA TCC CTG GGA GGT GTT TTG 94
Met Glu Ser Leu Leu Glv Glv Val Leu
1 5
CTG ATT TTC TCC CTT CAA GTG GAC TGG GTG AAG AGC CAA AAC ATA GAA 142
Leu Ile Leu Trp Leu Gln Asp Asp Val Lys Ser Val Gln Lys Glu Island
10 15 20 25
CAG AAT TCC GAG GCC CTG AAC ATT CAG GAG GGT AAA ACG GCC ACC CTG 190
Gln Asp Ser Glu Ala Leu Asn Gln Glu Glu Gls Lys Thr Ala Thr Leu
30 35 40
ACC TGC AAC TAT ACA AAC TAT TCT CCA GCA TAC TTA CAG TGG TAC CGA 238
Thr Cys Asn Tyr Thr Asn Tyr Pro Ser Ala Tyr Leu Gln Trp Tyr Arg
45 50 55
CAA GAT CCA GGA AGA GGC CCT GTT TTC TTG CTA CTC ATA CGT GAA AAT 286
Gln Asp Pro Glv Arg Glv Pro Val Phe Leu Leu Leu Isle Arg Glu Asn
60 65 70
GAG AAA GAA AAA AGG AAA GAA AGA CTG AAG GTC ACC TTT GAT ACC ACC 334
Glu Lys Glu Lys Arg Lys Glu Arg Leu Lys Asp Thr Thr Thr Thr Thr
75 80 85
CTT AAA CAG AGT TTG TT CAT ATC ACA GCC TCC CAG CT GAC GAC TCA 382
Leu Lvs Gln Ser Leu Phe His Thr Island Ser Ala Ser Gln Pro Ala Asp Ser
90 95 100 105
GCT ACC TAC CTC TGT GCT 400
Ala Thr Tyr Leu Cys Ala
110 x - INFORMATION FOR SEQ ID N 9
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 386 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
SEQUENCE CONSENSUS
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF DNA: IGR a 11
SEQUENCE V &alpha; 7
DESCRIPTION OF THE SEQUENCE
GCCTTCTGCA GACTCCAATG GCTCAGGAAC TGGGAATGCA GTGCCAGGCT 50
CGTGGTATCC TGCAGCAG ATG TGG GGA GTT TTG CTT CTT TAT GTT 95
Met Trp Gly Val Phe Leu Val Tyr Leu
1 5
TCC ATG AAG ATG GGA GGC ACT ACA GGA CAA AAC ATT GAC CAG CCC ACT 143
Ser Met Lys Met Gly Thr Thrly Gly Gln Asn Asp Asp Gln Pro Thr
10 15 20 25
GAG ATG ACA GCT ACG GAA GGT GCC ATT GTC CAG ATC AAC TGC ACG TAC 191
Glu Met Thr Ala Thr Glu Gly Ala Val Gln Island Asn Island Cys Thr Tyr
30 35 40
CAG ACA TCT GGG TTC AAC GGG CTG TTC TGG TAC CAG CAA CAT GCT GGC 239
Gln Thr Ser Gly Phe Asn Gly Leu Phe Trp Gln Tyr Gln His Ala Gly
45 50 55
GAA GCA CCC ACA TTT CTG TCT TAC AAT GTT CTG GAT GGT TTG GAG GAG 287
Glu Ala Pro Thr Phe Leu Ser Tyr Asn Val Leu Asp Gly Leu Glu Glu
60 65 70
AAA GGT CGT TT TCT TCA TTC CTT AGT CGG TCT AAA GGG TAC AGT TAC 335
Lys Gly Arg Ser Phe Ser Phe Leu Ser Arg Ser Lys Gly Tyr Ser Tyr
75 80 85
CTC CTT TTG AAG GAG CTC CAG ATG AAA GAC TCT GCC TCT TAC CTC TGT 383
Leu Leu Leu Lys Glu Leu Gln Met Lys Asp Ser Ala Ser Tyr Leu Cys
90 95 100 105
GCT 386
To the
XI - INFORMATION FOR SEQ IN N 10
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 383 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
DNA NAME: IGR has 12
SEQUENCE V &alpha; 22
DESCRIPTION OF THE SEQUENCE
TGTGACTTCT TCATGTTAAG GATCAAGACC ATTATTTGGG TAACACACTA 50
AAG ATG AAC TAT TCT CCA GGC TTA GTA TCT CTG ATA GTG TTA CTC 95
Asn Tyr Ser Pro Gly Leu Ser Ser Leu Leu Leu Leu Leu
1 5 10
CTT GGA AGA ACC CGT GGA GAT TCA GTG ACC AGC ATC GAA GGG CCA GTG 143
Leu Gly Arg Arg Thr Arg Gly Asp Ser Val Gln Gln Met Glu Gly Pro Val 15 20 25 30
ACT GTG TCA GAA GAG GCC TTC CTG ACT ATA AAC TGC ACG TAC ACA GCC 191
Thr Leu Ser Glu Glu Ala Phe Leu Thr Ile Asn Cys Thr Tyr Thr Ala
35 40 45
ACA GGA TAC CCT TCC CTT TTG TGG TAT GTC CAA TAT CCT GGA GAA GGT 239
Thr Gly Tyr Pro Ser Leu Phe Trp Tyr Val Gln Tire Pro Gly Glu Gly
50 55 60
CTA CAG GTG GTG CTG AAA GCC ACG AAG CGT GAT GAC AAG GGA AGC AAC 287
Leu Leu Leu Leu Leu Lys Ala Thr Lys Ala Asp Asp Lys Gly Ser Asn
65 70 75
AAA GGT TT GAA GCC ACA TAC CGT AAA GAA ACC ACT TCT TTC CAC TTC 335
Lys Gly Phe Glu Ala Thr Tyr Arg Lys Glu Thr Ser Ser Phe His Leu
80 85 90
GAG AAA GGC TCA GTT CAA GTG TCA GAC TCA GCG GTG TAC TTG TGT CGT 383
Glu Lys Gly Ser Gln Val Ser Val Ser Asp Ala Val Tyr Phe Cys Ala 95 100 105 .110
XII - INFORMATION FOR SEQ ID NO: 11
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 364 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
DNA NAME: IGR has 13
SEQUENCE V &alpha; 16
DESCRIPTION OF THE SEQUENCE
AATCCCGCCC GCCGTGAGCT TAGCTGGAGC ATG GCC TCT GCA CCC 46
Met Ala Ser Ala Pro
1 5
ATC TCG ATG CTT GCG ATG CTC TTC ACA TTG AGT AGG GGG AGG AGA GCT AGC 94
Ile Ser Met Leu Ala Met Leu Phe Thr Leu Ser Gly Leu Arg Ala Gln
10 15 20
TCA GTG CGT CAG CCG GAA GAT CAG GTc AAC GTT GCT GAA GGG AAT CCT 142
Ser Val Ala Gln Pro Glu Asp Gln Asn Val Val Ala Glu Gly Asn Pro
25 30 35
CTG ACT GTG AAA TGC ACC TAT TCA GTC TCT GGA AAC CCT TAT TT CTT 190
Leu Thr Val Lys CYS Thr Ser Ser Val Ser Gly Asn Pro Tyr Leu Phe
40 45 50
TGG TAT GTT CAA TAC CCC AAC CGA GGC GTG CAG TTG CTT CTG AAA TAC 238
Trp Tyr Val Gln Tyr Pro Asn Arg Gly Leu Gln Phe Leu Leu Lys Tyr
55 60 65
ATC ACA GGG GAT AAC CTG GTT AAA GGC AGC TAT GGC TT GAA GCT GAA 286
Thr Gly Asp Asn Leu Val Lys Gly Ser Tyr Gly Phe Glu Ala Glu 70 75 80 85
TTT AAC AAG AGC CAA ACC TCC TTC CAC CTG AAG AAA CCA TCT GCC CTT 334
Phe Asn Lys Ser Gln Ser Ser Phe His Leu Lys Pro Lys Ser Ala Leu
90 95 100
GTG AGC GAC TCC GCT TTG TAC TTc TGT GCT 364
Ser Ser Asp Ser Ala Leu Tyr Phe Cys Ala
105,110
XIII - INFORMATION FOR SEQ ID N 12
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 264 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: Ja 01
SEQUENCE
DESCRIPTION OF THE SEQUENCE
CCTTCAAGGA AAATTAAGGC AAATAGAATT GGGCTGGGGA GTTGCTACTT ATTAGTATTC 60
CTCCCACGTT CTAACCTAAT TATAAGGAGG TTGTTTTGGC CATGGGCAGT CATCTCAGGT 120
TTTGTTTTCC TGCTTTCCTC CCTAACCTCC ACCTGTCTTC CTAGAGGCCT GAGTCAAGGT 180
TATTGCAATA GCACTAAAGA CTGTGT ACC AAT GCA GGC AAA TCA ACC TT 233
Asn Thr Asn Ala Gly Lys Ser Thr Phe
1 5
GGG GAT GGG ACG ACT CTC ACT GTG AAG CCA 264
Gly Asp Gly Thr Thr Leu Thr Val Lys Pro
10 15
XIV - INFORMATION FOR SEQ ID NO: 13
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 277 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: Ja 02
SEQUENCE J &alpha;
DESCRIPTION OF THE SEQUENCE
AAGGACACAG ACTGCCTGCA TGAAGGCTGG AGCTGGGCCC AGGATGAGGA AAGGCCTCAG 60
GAAGGAAGGG CTGACACGAA ATAAGGAATA CCATGGCATT CATGAGATGT GCGTCTGAAT 120
CCTCTCTCTT GCCTGAGAAG CTTTAGCTTC CACCTTGAGA CACAAAACAT GTGGTTATGA 180
AGAGATGACA AGGTTTTTGT AAAAGAATGA GCCATTGTGG ATA GGC TT GGG AAT 234
Gly Phe Island Gly Asn i 5
GTG CTG CAT TGC GGG TCC GGC CAA ACT GTG ATT GTT TTA CCA 277
Val Leu His Cys Gly Ser Gly Thr Gln Val Ile Val Leu Pro
10 15 -XV - INFORMATION FOR SEQ ID.N 14
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
LENGTH: 58 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULES: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: Ja 03
SEQUENCE J &alpha;
DESCRIPTION OF THE SEQUENCE
AA ACC AGT GGC TCT AGG TTG ACC TT GGG GAA GGA ACA AGC CTC ACA 47
Thr Ser Gly Ser Arg Leu Thr Phe Gly Glu Gly Thr Gln Leu Thr
I 5 10 15
GTG AAT CCT GA 58
Val Asn Pro
XVI - INFORMATION FOR SEQ ID N- 15
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 60 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: Ja 04
SEQUENCE J &alpha;
DESCRIPTION OF THE SEQUENCE
TA GAT ACT GGA GGC VAT included AAA ACT ATC TT GGA GCA GGA ACA AGA CTA 47
Asp Thr Gly Gly Phe Lys Thr Ile Phe Gly Ala Gly Thr Arg Leu
1 5 10 15
TTT GTT AAA GCA A 60
Phe Val Lys Ala
XVII - INFORMATION FOR SEQ ID N-16
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 59 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: Ja 05
SEQUENCE J &alpha;
DESCRIPTION OF THE SEQUENCE
C CTA AGT GGG GCA AAC AAC GTC TTG TT GGG ACT GGA ACG AGA GTG 46
The Tht Oiy Ala Asn Asn Phe Phe Gly Tht Gly Tht Atg
i 5 10 15
ACC GTT CTT CCC T 59
Thr Val leu Pro
XVIII - INFORMATION FOR SEQ ID N 17
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 60 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T cells CHARACTERISTICS
NAME OF THE DNA: Ja 06
SEQUENCE J &alpha;
DESCRIPTION OF THE SEQUENCE
AT GGA GGA AGC CAA GGA AAT CTC ATC TTT GGA AAA GGC ACT AAA CTC 47
Gly Gly Ser Gln Gly Asn Leu Ile Phe Gly Lys Gly Thr Lys Leu
i 5 10 15
TGT GTT AAA CCA A 60
Ser Val Lys Pro
XIX - INFORMATION FOR SEQ ID N-18
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 56 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: Ja 07
SEQUENCE J &alpha;
DESCRIPTION OF THE SEQUENCE
GGA GCC AAT AGT AAG CTG ACA TT GGA AAA GGA ATA ACT CTG AGT GTT 48
Gly Ala Asn Ser Lys Leu Thr Phe Gly Lys Gly Thr Island Leu Ser Val 1 5 10 15
AGA CCA GA 56
Arg Pro
XX - INFORMATION FOR SEQ ID-N 19
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 57 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULE: cDNA for ARMm
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: Ja 08
SEQUENCE J &alpha;
DESCRIPTION OF THE SEQUENCE
GT GGT GGC TAC AAT AAG CTG ATT TT GGA GCA GGG ACC AGG CTG GCT 47
Ciy Oly Tyr Asn Lys The island Phe Oiy Ala Gly Tht Arg The eu Ala
i 5 10 15
GTA CAC CCA T 57
Val His Pro
XXI - INFORMATION FOR SEQ ID N- 20
CHARACTERISTICS OF THE SEQUENCES
TYPE: nucleotide and its corresponding protein
ponding
LENGTH: 50 base pairs
NUMBER OF BRINS: simple
CONFIGURATION: linear
TYPE OF MOLECULES: mRNA cDNA
ORIGIN
ORGANISM: human
CELL LINE: human T lymphocytes
CHARACTERISTICS
NAME OF THE DNA: Ja 09
SEQUENCE Jd
DESCRIPTION OF THE SEQUENCE
T GGA AAC AAG CTG GTC TTT GGC GCA GGA ACC ATT CTG AGA GTC AAG 46
Gly Asn Lys Leu Val Phe Gly Ala Gly Thr Ile Leu Arg Val Lys
10 15
i 5 10 15
TCC T 50
Ser
XXII - INFORMATION FOR SEQ ID N 21 to 25
TYPE OLIGONUCLEOTIDES
DESCRIPTION OF SEQUENCES
No 21 A (5'-GTTGCTCCAGGCCACAGCACTG)
N-22 poly C (5'-GCATGCGCGCGGCCGCGGAGG-14C)
N 23 B (5'-GTCCATAGACCTCATGTCCAGCACAG)
N 24 C (5'-ATACACATCAGAATTCTTACTTTG)
N 25 D (5'-GTCACTGGATTTAGAGTCT)

Claims (23)

 1. Nucleotide sequences encoding variable regions of &alpha; human T cell receptors, corresponding to cDNAs comprising nucleotide sequences selected from any one of  a - Vi segments corresponding to one of the sequences SEQ ID N 1 to 11, and  b - segments Jd corresponding to one of the sequences SEQ ID No. 12 to 20 and the sequences which differ from it by one or more nucleotides.  2. Sequences according to claim 1 coding for variable regions of the human T cell receptor chains, corresponding to CDNA comprising nucleotide sequences chosen from any one of the segments V g corresponding to one of the sequences SEQ ID N 1 to 10, and the sequences which differ therefrom by one or more nucleotides.  3. Sequences according to claim 1 encoding variable regions of the human T cell receptor chains, corresponding to CDNA comprising nucleotide sequences chosen from any of the segments J i corresponding to one of the sequences SEQ ID N 12 to 20, and the sequences which differ from it by one or more nucleotides.  4. Nucleotide sequences coding for variable regions of &alpha; human T cell receptors corresponding to cDNAs corresponding to all or part of the nucleotide sequences selected from any of the V & alpha segments corresponding to one of SEQ ID N 1 to 5 sequences, and sequences which differ by one or two nucleotides.  5. Nucleotide sequences encoding human T-cell receptor variable region A corresponding to cDNAs corresponding to all or part of the nucleotide sequences selected from any of the Vi segments corresponding to one of the sequences  1 to 467 of SEQ ID N 6  1 to 77 of SEQ ID N 7  1 to 151 of SEQ ID N 8  291 to 386 of SEQ ID N 9  1 to 260 of SEQ ID N 10 and the sequences which differ therefrom by one or two nucleotides.  6. Nucleotide sequences coding for variable regions of human T-lymphocyte receptor chains corresponding to cDNAs corresponding to all or part of the nucleotide sequence corresponding to SEQ ID No. 11 and which comprise nucleotide 108.  7. Nucleotide sequences encoding Q chain variable regions (human T cell receptors corresponding to cDNAs corresponding to all or part of the nucleotide sequences selected from any one of the segments corresponding to one of the SEQ ID sequences N 12 to 20 and the sequences differing therefrom by one or two nucleotides.  8. Peptides encoded by any of the nucleotide sequences as defined in any one of claims 1 to 7, as well as alleles and derivatives thereof that have the same function.  9. Peptides constituted by or comprising a peptide sequence encoded by all or part of the sequence 108 to 364 of SEQ ID N-11.  10. Vectors. expression system comprising a DNA sequence encoding one of the peptides as defined in claim 8 or claim 9.  11. Worms transformed with a vector according to claim 10.  12. Antibodies directed against an antigenic determinant of one of the peptides defined in claim 8 or claim 9.  An antibody according to claim 12 which is a monoclonal antibody or a fragment thereof.  14. Fab, Fab 'or Fab' fragment 2 of a monoclonal antibody according to claim 13 and derivatives thereof.  15, -Derivatives of a monoclonal antibody or a fragment thereof according to claim 13 or 14 which are bound to a detectable label and / or at least one therapeutic molecule.  Hybridomas producing an antibody according to claim 13.  17. A diagnostic composition comprising one or more monoclonal antibodies, fragments or derivatives thereof as defined in any one of claims 13 to 15.  18. Therapeutic composition comprising one or more monoclonal antibodies, fragments or derivatives thereof as defined in one of claims 13 to 15.  19. The composition of claim 18 comprising derivatives containing a cytotoxic molecule or a radioisotope.  20. Therapeutic composition comprising at least one of the peptides defined in claim 8 or claim 9.  21. A therapeutic composition comprising antisense oligonucleotides corresponding to any one of the sequences of a segment V i or in any of the sequences of a segment J i as defined in any one of claims 4 to 7.  22. Use, as primers for DNA amplification, of nucleotide sequences of at least about 17 nucleotides included in any one of the sequences of a VO segment (or any of the sequences of a J & alpha segment as defined in any one of claims 4 to 7.  23. Use, as a detection probe, of nucleotide sequences of at least about 10 nucleotides included in any of the sequences of a V-segment or in any of the sequences of a J & alpha segment, such as defined in any one of claims 4 to 7.