ANTI-IL17 ANTIBODIES

Description

FIELD OF THE INVENTION

[0001] The present invention relates to the identification of biological markers of rheumatoid arthritis (RA). The invention describes a method for determining the level of these new biomarkers, new antibodies as well as their therapeutic, diagnostic or prognostic applications. PREVIOUS ART

[0002] Autoimmune diseases are generally characterized by the triggering of an immune reaction, which results in an inflammatory reaction, against substances and / or tissues normally present in the individual. They are often, but not systematically, highlighted by the detection of autoantibodies directed against "self" antigens.

[0003] These chronic autoimmune and inflammatory diseases are heterogeneous pathologies, for which there is a high variability in phenotypes and responses to treatments. Thus, the prognosis of these chronic autoimmune and inflammatory diseases is likely to vary considerably within individuals who are nevertheless considered to have the same pathology.

[0004] In particular, rheumatoid arthritis is a chronic inflammatory degenerative disease, characterized by joint damage that is often bilateral and symmetrical, progressing in flare-ups towards deformation and destruction of the affected joints. Symptomatic treatment involves non-steroidal anti-inflammatory drugs and corticosteroids. Nowadays, methotrexate is the standard treatment. "Basic" treatments for the disease are actively sought, particularly by identifying the cytokines and cells involved in the inflammatory process of the disease. These treatments use specific inhibitors of the cytokines and cells involved. Examples include Etanercept ®< and Infliximab ®<, TNF (Tumor Necrosis Factor) inhibitors.Other treatments involve interleukin antagonists such as IL-1 inhibitors (Anakinra ®< ), IL-6 receptor inhibitors (MrA ®< ) and anti-CD20 (Rituximab ®< ). These compounds reduce inflammation and slow the progression of the disease.

[0005] However, there is a wide variation in response between rheumatoid arthritis patients, who do not respond similarly to these treatments. It is estimated that up to 30% of patients do not respond at all to the biotherapies listed above.

[0006] Jouvenne P. et al., Scand. J. Immunol., 1997, 46(4): 413-418 reports a follow-up study of the evolution of patients suffering from RA with and without bone destruction which shows that high levels of neutralizing antibodies directed against IL-1α are associated with a better prognosis of RA and concludes that natural autoantibodies against IL-1α can protect against or delay the onset of RA with bone destruction and act in a dose-dependent manner for the evolution of the disease.

[0007] IL-17 has been shown to play a major role in inflammatory diseases, autoimmune diseases, certain infections, and cancer. Thus, this cytokine is now considered a potential therapeutic target for many diseases (Miossec et al., N. Engl. J. Med. 2009, 361(9): 888-898). The two major forms of this cytokine are called IL-17A and IL-17F. IL-17A induces the production of numerous cytokines and chemokines, such as IL-6, G-CSF, IL-1β, and IL-8. In rheumatoid arthritis, IL-17 is present at sites of inflammation and acts by amplifying the inflammatory effects of other compounds such as TNF, making it an important player in the physiology of the disease. However, circulating levels of IL-17 in the plasma of patients are often very low, and very high heterogeneity of circulating IL-17 levels is observed between patients.

[0008] IL-17A inhibitors, including antibodies, have been proposed for the treatment of inflammatory diseases, and various clinical trials are underway (Miossec and Kolls, Nat. Rev. Drug Discov. 2012, 11(10): 763-776). Initial results indicate that it is difficult to predict how patients will respond to these treatments, as there is considerable individual variation.

[0009] In some patients, the role of IL-17 in the inflammatory process is significant, while in other patients it is very minor. A major effort must now be made to pre-select patients whose inflammatory state is directly dependent on IL-17, because they are the ones who would benefit from this type of treatment.

[0010] WO2015 / 186106 teaches in particular processes in vitroimplementing the level of functional IL-17 (IPDL) as a biomarker, to determine the chances of response of a patient with a chronic inflammatory state to the administration of an anti-IL17 antibody.

[0011] Thus, there remains a need to identify new markers of rheumatoid arthritis. There also remains a need to identify new methods and tools for the diagnosis, prognosis and / or treatment of this chronic autoimmune and inflammatory disease. SUMMARY OF THE INVENTION

[0012] The present invention describes a biomarker for rheumatoid arthritis, based on the detection in a biological sample of an anti-IL-17 autoantibody, which can be determined either at a total level or in complex with Interleukin-17.

[0013] The present invention therefore relates to a method in vitroto assess the prognosis of rheumatoid arthritis in an individual, including the following steps: a) determining (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from said individual, and b) comparing the level of autoantibodies and / or complex determined in step a) with a reference value, said comparison being indicative of the prognosis of rheumatoid arthritis in said individual.

[0014] The above methods also make it possible to identify particular populations of individuals, such as particular populations of individuals suffering from rheumatoid arthritis, for whom an active ingredient inhibiting IL-17 is administrable.

[0015] Knowledge of this new biomarker allows in particular the following diagnostic and / or prognostic applications: (i) determining the risk of severity, in particular bone destruction, in an individual suffering from rheumatoid arthritis; (ii) determining the chances of response of an individual suffering from rheumatoid arthritis to a treatment comprising the administration of an active ingredient inhibiting IL-17, such as an anti-IL17 antibody. (iii) determining the efficacy of a treatment or prevention of bone destruction in an individual suffering from rheumatoid arthritis, said treatment or said prevention consisting of the administration of an active ingredient inhibiting IL-17, such as an anti-IL-17 antibody.

[0016] In particular, the present invention describes a method in vitro of antibody selection for treating (i) rheumatoid arthritis, and / or (ii) bone destruction associated with said disease, comprising the following steps: a)determining the presence of (i) an anti-IL-17 autoantibody and / or (ii) an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from an individual evaluated by said method in vitro or according to claim 1; b) selecting an anti-IL-17 autoantibody or a cell producing an anti-IL-17 autoantibody from a biological sample of said individual; said autoantibody being capable of treating said disease, and / or said bone destruction.

[0017] The present invention also relates to a method in vitro to distinguish destructive rheumatoid arthritis from non-destructive rheumatoid arthritis in an individual, comprising the following steps: a) determining (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from said individual, and b)comparing the level of autoantibodies and / or complex determined in step a) with a reference value, said comparison being indicative of destructive or non-destructive rheumatoid arthritis in said individual.

[0018] The present invention also relates to an isolated anti-IL-17 antibody for the treatment or prevention of (i) destructive rheumatoid arthritis and / or (ii) rheumatoid arthritis in an individual not producing anti-IL-17 autoantibodies, or bone destruction associated with any of groups (i) or (ii).

[0019] The present invention also relates to the use of an isolated anti-IL-17 antibody for the preparation of a medicament for the treatment or prevention of (i) destructive rheumatoid arthritis and / or (ii) rheumatoid arthritis in an individual not producing anti-IL-17 autoantibodies, or bone destruction associated with any of groups (i) or (ii). LEGEND OF THE FIGURES

[0020] Figure 1 : Principle of detection of anti-IL-17 autoantibodies and IL-17 / anti-IL-17 antibody complexes.Anti-IL-17 autoantibodies are detected in plasma by a competitive ELISA (A). IL-17 and plasma are combined to promote the binding of IL-17 and anti-IL-17 antibody. Anti-IL-17 and irrelevant antibodies were used as positive and negative controls, respectively. A detection anti-IL-17 antibody was then added. The mixture is then transferred to plates containing the capture anti-human IL-17 antibody. The [IL-17 / anti-IL-17 autoantibodies] complexes are detected in plasma by an indirect ELISA (B). In a 96-well plate containing the anti-IL-17 capture antibody, plasma pre-incubated with horse serum is added. The positive control immune complex (IC) was formed in vitrofrom 500 ng of IL-17 and 5 µg / ml of human anti-human IL-17 antibody. A goat anti-human IgG Fc fragment antibody (anti-human IgG Fc) conjugated to peroxidase was used for the revelation. Figure 2 : Detection of IL-17 / anti-IL-17 antibody complexes in the plasma of patients with destructive and non-destructive rheumatoid arthritis, compared to healthy donors.

[0021] An eight-point standard curve is determined from serial dilutions (2-fold) and tested with the formed positive control in vitro from 500 ng of IL-17 and 5 µg / ml of human anti-human IL-17 antibody (A).

[0022] Immune complexes (ICs) were detected in the plasma of 30 healthy donors and 60 patients with rheumatoid arthritis (RA) with or without bone destruction. For each plasma sample, a standard curve of serial dilutions (2-fold) was tested to determine the immune complex titers. (B).

[0023] The mean level of immune complexes [IL-17 / anti-IL-17 autoantibodies] in the non-destructive RA vs. destructive RA groups are shown. (C). The Mann-Whitney test shows a statistically significant difference (p value) (* < 0.05; *** < 0.001). Error bars represent the standard deviation. DETAILED DESCRIPTION OF THE INVENTION

[0024] Surprisingly and unexpectedly, the inventors identified the presence of anti-IL-17 autoantibodies in individuals with rheumatoid arthritis.

[0025] Also surprisingly, the inventors showed that the production of anti-IL-17 autoantibodies and / or immune complexes [IL-17 / anti-IL-17 autoantibodies] was more common in individuals with non-destructive rheumatoid arthritis than those with destructive rheumatoid arthritis.

[0026] This negative relationship between the presence of anti-IL-17 autoantibodies and the phenomenon of bone destruction in this chronic autoimmune and inflammatory disease suggests that these autoantibodies and complexes [IL-17 / anti-IL-17 autoantibodies] protect against bone destruction, by neutralizing all or part of the function of Interleukin 17 (IL-17).

[0027] IL-17 or IL17 refers to interleukin-17 as identified in 1993 by Rouvier's team et al.It was renamed IL-17A after the identification of new family members, designated IL-17B to IL-17F. IL-17 is a homodimeric glycoprotein of 155 amino acids, weighing 35 kDa. IL-17 is secreted by CD4+ and CD8+ lymphocytes and is involved in the coordination of local tissue inflammation, notably via the induction of pro-inflammatory cytokine secretion and inducing neutrophil mobilization (Kolls and Linden, Immunity 2004, 21(4):467-476).

[0028] Indeed, IL-17 induces the secretion of pro-inflammatory factors by its numerous target cells, each type of target cell being specialized in the production and secretion of one or more cytokines or chemokines.

[0029] The present invention relates in particular to IL-17A, but can also be applied to other members of the family: IL-17B, IL-17C, IL-17D, IL-17E and in particular IL-17F which has the highest rate of sequence identity with IL-17A. By analogy, antibodies, anti-IL-17A autoantibodies, and complexes comprising said autoantibodies are particularly considered, although those specifically directed against the other members are also relevant from a clinical point of view (diagnosis, prognosis, and / or therapy).

[0030] It was certainly known that chronic autoimmune diseases could be characterized by the presence of autoantibodies. It was also known that Interleukin 17 (IL-17) could constitute a therapeutic target for a growing number of inflammatory diseases.

[0031] Thus, clinical trials for the treatment of rheumatoid arthritis using anti-IL17 antibodies have already been reported, notably by Hueber et al. (Sci Transl Med. 2010;2:52ra72) and Genovese et al. (Arthritis Rheumatol. 2014;66:1693-1704). However, the results from these clinical trials are heterogeneous.

[0032] Without wishing to be bound by theory, the inventors are of the opinion that this heterogeneity is linked to the presence, at variable levels, of anti-IL17 autoantibodies and / or immune complexes [IL-17 / anti-IL-17 autoantibodies].

[0033] In particular, the results obtained (see examples) on biological samples from patients suggest that, in non-destructive rheumatoid arthritis, these anti-IL17 autoantibodies are present in excess and bind interleukin IL-17 to form more [IL-17 / anti-IL-17 autoantibodies] complexes; the observed consequence is thus a decrease or even an absence of detectable levels of free bioactive IL-17.

[0034] The detection of anti-IL17 autoantibodies and / or immune complexes [IL-17 / anti-IL-17 autoantibodies] in these individuals therefore represents a biomarker of interest for predicting the prognosis of these chronic autoimmune and inflammatory diseases, in particular for predicting bone destruction associated with these diseases, but also for predicting a response to treatment in this context.

[0035] We understand by " bone destruction» a progressive disappearance of bone tissue due to the patient's chronic inflammatory state.

[0036] We understand by " destructive rheumatoid arthritis ", the occurrence of rheumatoid arthritis in an individual, for which bone destruction is associated. Conversely, " non-destructive rheumatoid arthritis » , the occurrence of rheumatoid arthritis in an individual, for which bone destruction is not associated.

[0037] In view of the experimental data, a " non-destructive rheumatoid arthritis » is statistically likely to correspond to rheumatoid arthritis associated with the production of anti-IL17 autoantibodies.

[0038] Conversely, a " destructive rheumatoid arthritis » is statistically likely to correspond to rheumatoid arthritis not associated with the production of anti-IL17 autoantibodies, or for which the production of anti-IL17 autoantibodies is insufficient.

[0039] The occurrence of destructive or non-destructive rheumatoid arthritis can also be determined from the Larsen score, and in particular the wrist Larsen score. A Larsen score above 2 is generally considered characteristic of destructive rheumatoid arthritis. A Larsen score between 0 and 1 generally corresponds to non-destructive rheumatoid arthritis. The meaning and determination of a Larsen score, in particular a wrist Larsen score, is part of the general knowledge of the person skilled in the art.

[0040] In particular, the term " determination of the production of anti-IL17 autoantibodies”, (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex.

[0041] The term " antibody» is used here in its most general sense, and specifically covers monoclonal antibodies (including whole monoclonal antibodies), polyclonal antibodies (e.g., bispecific antibodies), and antibody fragments provided they exhibit the desired biological activity.

[0042] The term " antibody fragment » refers to antibody fragments comprising a portion of a complete antibody, typically the portion responsible for binding to the antigen or its variable domain. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibodies; and multispecific antibodies formed from antibody fragments.

[0043] The term " autoantibodies » refers to an antibody produced by the host's own immune system and directed against one or more of its proteins.

[0044] The term " individual» includes any individual, human or non-human (preferably human) susceptible to developing a chronic autoimmune or inflammatory disease.

[0045] According to a first embodiment, the invention relates to a method in vitro to assess the prognosis of rheumatoid arthritis in an individual, including the following steps: a) determining (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from said individual, and b) comparing the level of autoantibody and / or complex determined in step a) with a reference value, said comparison being indicative of the prognosis of rheumatoid arthritis in said individual. rheumatoid arthritis may be: destructive rheumatoid arthritis; non-destructive rheumatoid arthritis; rheumatoid arthritis in an individual producing anti-IL-17 autoantibodies; or rheumatoid arthritis in an individual not producing anti-IL-17 autoantibodies.

[0046] More specifically, the process in vitro as defined above can be implemented for (i) determining the risk of bone destruction in an individual suffering from rheumatoid arthritis; and / or (ii) determining the chances of response of an individual suffering from rheumatoid arthritis to a treatment comprising the administration of an active ingredient inhibiting IL-17 (in particular an anti-IL-17 antibody); and / or (iii) determining the efficacy of a treatment or prevention of bone destruction in an individual suffering from rheumatoid arthritis, said treatment or said prevention consisting of the administration of an active ingredient inhibiting IL-17 (in particular an anti-IL-17 antibody).

[0047] Thus, according to these particular embodiments, the comparison in step b) is indicative of said risk of bone destruction, of said chances of response to said treatment, and / or of the effectiveness of said treatment or of said prevention of bone destruction.

[0048] The present disclosure describes a method in vitroselection of an individual in whom the presence of anti-IL-17 autoantibodies reflects the presence in a biological sample (including blood) of said individual of cells producing this antibody. This presence of cells producing anti-IL-17 autoantibodies allows their isolation from said sample, and in particular from a blood sample.

[0049] The use of current laboratory techniques then makes it possible to isolate the so-called anti-IL-17 autoantibody, particularly for therapeutic use.

[0050] In particular, this process in vitro evaluation can be implemented in a process in vitro of antibody selection for treating (i) rheumatoid arthritis and / or (ii) bone destruction associated with said disease, comprising the following steps: a) determining the level of (i) an anti-IL-17 autoantibody and / or (ii) an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample of an individual evaluated according to the methods described above, and / or claim 1; b) selecting an anti-IL-17 autoantibody or a cell producing an anti-IL-17 autoantibody from a biological sample of said individual; said autoantibody being capable of treating said disease, and / or said bone destruction.

[0051] Biological samples of the stages a) And b) of the said selection process may be identical or different. The step b) selection can be carried out from any biological sample likely to contain the said anti-IL-17 autoantibody or the said producing cell, which includes: serum, plasma, blood and primary or secondary lymphoid organs.

[0052] Thus, the invention relates to a method in vitro of antibody selection for treating (i) rheumatoid arthritis, and / or (ii) bone destruction associated with said disease, comprising the following steps: a) determine the level of (i) an anti-IL-17 autoantibody and / or (ii) an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from an individual with rheumatoid arthritis b) compare the level of autoantibody and / or complex determined in step a) with a reference value; c) selecting an anti-IL-17 autoantibody or a cell producing an anti-IL-17 autoantibody from a biological sample of said individual; said autoantibody being capable of treating said disease, and / or said bone destruction.

[0053] In the context of the said procedures in vitroantibody selection, the biological sample may in particular come from an individual preferably suffering from non-destructive rheumatoid arthritis.

[0054] According to a second embodiment, the invention relates to a method in vitro to distinguish destructive rheumatoid arthritis (RA) from non-destructive rheumatoid arthritis in an individual, comprising the following steps: a) determining (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from said individual, and b) comparing the level of autoantibodies and / or complex determined in step a) with a reference value, said comparison being indicative of destructive or non-destructive rheumatoid arthritis (RA) in said individual.

[0055] According to a third embodiment, the invention relates to an isolated anti-IL-17 antibody, for the treatment or prevention of (i) destructive rheumatoid arthritis and / or (ii) non-destructive rheumatoid arthritis and / or (iii) rheumatoid arthritis in an individual producing anti-IL-17 autoantibodies and / or (iv) rheumatoid arthritis in an individual not producing anti-IL-17 autoantibodies, and / or (v) bone destruction with respect to any of groups (i) to (iv).

[0056] According to a fourth embodiment, the invention relates to an isolated human anti-IL-17 antibody, obtained from a biological sample of an individual suffering from rheumatoid arthritis and producing anti-IL-17 autoantibodies.

[0057] An anti-IL-17 antibody isolated according to the invention is preferably an antibody isolated from the method in vitroselection of the invention, and / or an antibody isolated from a biological sample of an individual suffering from rheumatoid arthritis, and preferably suffering from non-destructive rheumatoid arthritis.

[0058] In particular, said anti-IL17 antibody can be obtained according to a method comprising the following steps: a) preparing a composition enriched in lymphocytes producing anti-IL17 antibodies from a sample originating from an individual suffering from rheumatoid arthritis, and preferably suffering from non-destructive rheumatoid arthritis, b) selecting at least one B lymphocyte producing anti-IL17 antibodies, or at least one clone of B lymphocytes producing anti-IL17 antibodies, and c) obtaining said anti-IL17 antibody.

[0059] In some embodiments, the sample from which the composition enriched in anti-IL17 antibody-producing lymphocytes is prepared, in step a), is a sample from human blood, which includes whole blood and a cell-enriched fraction of the blood such as a peripheral blood mononuclear cell-enriched fraction and a lymphocyte-enriched fraction.

[0060] Step b) can be carried out using any technique well known to those skilled in the art.

[0061] Step c) can be carried out by culturing the B lymphocyte(s) selected in step b), then purifying the anti-IL17 antibody thus produced.

[0062] Most commonly, the B lymphocyte(s) selected in step b) are then immortalized in the form of a cell line producing the said anti-IL17 antibody. The cells in the line are then cultured and the anti-IL17 antibody thus produced is then purified. Processes in vitro

[0063] For the purpose of carrying out the processes in vitro described above, we mean by " reference value ", a level of anti-IL-17 autoantibodies and / or (ii) level of [IL-17 / anti-IL-17 autoantibodies] complex determined in individuals / patients affected by rheumatoid arthritis, and for whom the occurrence or not of bone destruction is possibly known, or for whom a phenomenon of bone destruction has been observed where appropriate.

[0064] In general, said reference value is an average value measured from a biological sample of a plurality of individuals affected by rheumatoid arthritis, and whose occurrence (or not) of bone destruction, or whose production of said levels of anti-IL-17 autoantibodies and / or (ii) level of [IL-17 / anti-IL-17 autoantibodies] complex is known.

[0065] For the purposes of implementing the methods according to the invention, it is possible to choose as " reference value » an average value determined or measured in patients affected by rheumatoid arthritis, and in whom the occurrence or not of bone destruction is observed.

[0066] In some embodiments of the method, the reference value is a value determined in individuals without bone destruction and / or in individuals whose rheumatoid arthritis is associated with the production of anti-IL-17 autoantibodies and / or [IL-17 / anti-IL-17 autoantibodies] complex. In these embodiments, a patient tested according to the method of the invention will be classified as " at reduced or no risk of bone destruction » when the (i) level of anti-IL-17 autoantibodies and / or (ii) level of [IL-17 / anti-IL-17 autoantibodies] complex is higher than the said reference value.

[0067] In some embodiments of the method, the reference value is a value determined in patients with bone destruction and / or in individuals whose rheumatoid arthritis is not associated with the production of anti-IL-17 autoantibodies and / or [IL-17 / anti-IL-17 autoantibodies] complex.

[0068] In these embodiments, a patient tested according to the method of the invention will be classified as “ at moderate or high risk of bone destruction » when the (i) level of anti-IL-17 autoantibodies and / or (ii) level of [IL-17 / anti-IL-17 autoantibodies] complex is lower than the said reference value.

[0069] In certain embodiments of the method, the reference value is a value determined in the same patient, before or after administration of an active ingredient to said patient, or before or after bringing the biological sample into contact with said active ingredient. This embodiment can be advantageously used to determine: the chances of response of an individual suffering from rheumatoid arthritis to a treatment comprising the administration of an active ingredient inhibiting IL-17; and / or the effectiveness of a treatment or prevention of bone destruction in an individual suffering from said disease, said treatment or said prevention consisting of the administration of an active ingredient inhibiting IL-17.

[0070] The administration step of an active ingredient, or candidate compound, will be carried out over a period suitable for judging the effectiveness of said compound, this period ranging from one day to several months, and being modelled on the usual administration periods for this type of compound.

[0071] Thus, according to a particular embodiment, the invention relates to a method in vitro to assess the prognosis of rheumatoid arthritis in an individual, to determine: the chances of response of said individual to a treatment comprising the administration of an active ingredient inhibiting IL-17; and / or the effectiveness of a treatment or prevention of bone destruction of said individual, said treatment or said prevention consisting of the administration of an active ingredient inhibiting IL-17; and comprising the following steps: a)determining (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from said individual prior to said administration; b) administering to the individual the said IL-17 inhibitor active ingredient; c) determining (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from said individual after said administration; and d) comparing the level of autoantibodies and / or complex determined in steps a) and c) with a reference value, said comparison being indicative of said chances of response and / or the effectiveness of said treatment or prevention.

[0072] It is understood that said IL-17 inhibitory active ingredient (which is preferably an anti-IL-17 antibody) is considered effective when the value determined in step a) is greater than the value determined in step c).

[0073] In still other embodiments, the reference value is a so-called " threshold » or « cut-off » , which is determined from (i) values ​​determined in patients " at reduced or no risk " and (ii) values ​​determined in patients " at moderate or significant risk » . In these embodiments, a patient tested according to the method of the invention will be classified as “ at reduced or no risk of bone destruction » when the (i) level of anti-IL-17 autoantibodies and / or (ii) level of [IL-17 / anti-IL-17 autoantibodies] complex measured for this patient is higher than the reference value.

[0074] A reference value " threshold » or « cut-off» can be easily determined by a person skilled in the art using his general knowledge. A reference value "threshold" or "cut-off" can be determined, for example, as described by Limmathurotsakul et al. (2011, Clin. Infect. Dis., Vol. 52: 1024-1028).

[0075] It is understood that for the implementation of all these procedures, the biological sample was taken from the individual tested, and that it is a sample of whole blood, plasma, serum, synovial fluid, cerebrospinal fluid, pleural fluid, or peritoneal fluid of said individual.

[0076] The said processes in vitro can also be carried out on any type of cell capable of producing said autoantibodies, which includes B lymphocytes from individuals with a chronic autoimmune or inflammatory disease, for example rheumatoid arthritis.

[0077] Where the biological sample comprises or consists of cells, these methods will preferably be characterised in that the cells used are primary cell cultures, in particular cells taken from the individual whose biological sample is being tested.

[0078] According to a particular embodiment of these methods in vitro, step a) consists of determining the level of an anti-IL-17 autoantibody in said biological sample.

[0079] According to another particular embodiment of these methods in vitro, the stage a) consists of determining the level of a complex [IL-17 / anti-IL-17 auto-antibodies] in the said biological sample.

[0080] In particular, the biological sample can be chosen from: whole blood, plasma, and serum.

[0081] According to a preferred embodiment, the anti-IL-17 autoantibody is an anti-IL-17A autoantibody; and the [IL-17 / anti-IL-17 autoantibody] complex is an [IL-17A / anti-IL-17A autoantibody] complex.

[0082] Thus, the anti-IL-17 autoantibody may preferably be a human autoantibody directed against human interleukin IL-17A.

[0083] Determination of the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample can be performed by any test known to those skilled in the art. In a non-exhaustive manner, tests for the determination of autoantibodies are referenced in Aggarwal et al (Best Practice & Research Clinical Rheumatology 28 (2014) 907-920).

[0084] For example, the determination of the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex can be carried out according to one of the following methods: immunoelectrophoresis, counterelectrophoresis, gel immunodiffusion, immunoagglutination, immunofluorescence, ELISA, nephelometry and immunoblotting.

[0085] According to one embodiment, step a) is implemented in the form of an ELISA test. Immunological quantification tests in the form of ELISA tests are known to those skilled in the art. They can in particular be implemented in the form of competitive ELISAs (see examples and Figure 1A ) or indirect ELISA (see Figure 1B ).

[0086] According to one sub-embodiment, a competitive ELISA assay can be performed to determine the level of an anti-IL-17 autoantibody in a sample.

[0087] According to one sub-embodiment, an indirect ELISA test can be performed to determine the level of an [IL-17 / anti-IL-17 autoantibody] complex in a sample.

[0088] Thus, a competitive ELISA test to determine the level of an anti-IL-17 autoantibody in a sample may include the following steps: a) optionally bringing into contact a sample likely to contain an anti-IL17 autoantibody, in the presence of an agent likely to inhibit the interaction of rheumatoid factor possibly present with said autoantibodies (for example a serum from another organism); b) contacting said sample with an interleukin IL-17 for a time sufficient for said anti-IL-17 autoantibodies to interact with said interleukin; c)contacting said sample with an exogenous antibody directed against said interleukin for a time sufficient for said exogenous antibody to compete with said anti-IL-17 autoantibodies to interact with said interleukin; d) determining the level of anti-IL-17 autoantibodies in said sample from the level of exogenous antibody interacting with said interleukin.

[0089] In particular, said exogenous antibody may be in labeled form, in order to facilitate the determination of the level of autoantibodies in step d).

[0090] Step d) may further include a step of fixing said biological sample on a support on which anti-IL17 antibodies are fixed.

[0091] An indirect ELISA test to determine the level of an [IL-17 / anti-IL-17 autoantibody] complex in a sample may include the following steps: a)contacting a sample likely to contain an [IL-17 / anti-IL-17 autoantibody] complex with a support on which anti-IL17 antibodies are fixed, for a time sufficient for said complexes to interact with said antibody.; b) contacting said sample with an exogenous antibody directed against said anti-IL-17 autoantibody, for a time sufficient for said exogenous antibody to interact with said anti-IL-17 autoantibody. c) determining the level of [IL-17 / anti-IL-17 autoantibody] complex in said sample from the level of exogenous antibody interacting with said autoantibody.

[0092] In particular, said exogenous antibody may be in labeled form, in order to facilitate the determination of the level

[0093] The said samples may in particular be blood samples, or blood derivatives, such as plasma or serum.

[0094] According to one embodiment, a method in vitro according to the invention may further comprise a step of determining, in a biological sample of said individual, the level of a polypeptide chosen from: cytokines IL-17A, IL-17F, IL-25, IL-23, transcription factors, including RORγt, and interleukin IL-17 receptors such as IL-17RA, IL-17RB, IL-17RC, autoantibodies.

[0095] Thus, according to a particular embodiment, a method in vitro according to the invention may comprise a step of determining, in a biological sample of said individual, the level of a polypeptide chosen from: the cytokines IL-17A, IL-17F, IL-25, IL-23, the transcription factor RORγt, and the receptors IL-17RA, IL-17RB, IL-17RC, the autoantibodies directed against IL-1α, the autoantibodies directed against IL-8 and / or the autoantibodies directed against osteopontin. Pharmaceutical composition and therapeutic applications

[0096] Anti-IL-17 antibodies per se, and for the treatment or prevention of autoimmune or chronic inflammatory diseases are described below.

[0097] These anti-IL-17 antibodies can be used as drugs or even for the preparation of a drug.

[0098] The identification of anti-IL-17 autoantibodies and [IL-17 / anti-IL-17 autoantibodies] complexes in patients with destructive and non-destructive rheumatoid arthritis also allows consideration of the administration of anti-IL-17 antibodies to specific subgroups of individuals with: Destructive rheumatoid arthritis; non-destructive rheumatoid arthritis; rheumatoid arthritis associated with the production of anti-IL-17 autoantibodies; rheumatoid arthritis not associated with the production of anti-IL-17 autoantibodies; and / or bone destruction associated with any of the preceding subgroups.

[0099] Thus, the present invention relates to an isolated anti-IL-17 antibody for the treatment or prevention of (i) destructive rheumatoid arthritis and / or (ii) non-destructive rheumatoid arthritis and / or (iii) rheumatoid arthritis in an individual producing anti-IL-17 autoantibodies and / or (iv) rheumatoid arthritis in an individual not producing anti-IL-17 autoantibodies, and / or (v) bone destruction associated with any of groups (i) to (iv).

[0100] The present invention also relates to the use of an isolated anti-IL-17 antibody for the preparation of a medicament for the treatment or prevention of (i) destructive rheumatoid arthritis and / or (ii) non-destructive rheumatoid arthritis and / or (iii) rheumatoid arthritis in an individual producing anti-IL-17 autoantibodies and / or (iv) rheumatoid arthritis in an individual not producing anti-IL-17 autoantibodies, and / or (v) bone destruction associated with any of groups (i) to (iv).

[0101] Preferably, the subgroups of individuals to be considered in the context of administration of anti-IL-17 antibodies and / or preparation of a drug are those suffering from: Destructive rheumatoid arthritis; rheumatoid arthritis not associated with the production of anti-IL-17 autoantibodies; and / or bone destruction associated with any of the preceding subgroups.

[0102] The said subgroups of individuals may in particular be those evaluated by any of the processes in vitro evaluation according to the invention.

[0103] The said anti-IL-17 antibodies according to the invention may in particular be those selected by any of the methods in vitro selection according to the invention.

[0104] An anti-IL-17 antibody according to the invention may be an anti-IL-17 antibody obtained from a biological sample of an individual producing anti-IL-17 autoantibodies.

[0105] In particular, said anti-IL-17 antibody is an isolated human anti-IL-17 antibody obtained from a biological sample of an individual producing anti-IL-17 autoantibodies.

[0106] An anti-IL-17 antibody may also be derived from a cell producing anti-IL-17 autoantibodies; said producing cell (for example a B lymphocyte and / or a hybridoma) being obtained from a biological sample of an individual producing said anti-IL-17 autoantibodies.

[0107] The said individual from whom the said anti-IL-17 autoantibody or the said producing cell is isolated, suffers from rheumatoid arthritis.

[0108] According to one embodiment, the anti-IL-17 antibody is an isolated human anti-IL-17 antibody obtained from a biological sample of an individual suffering from rheumatoid arthritis and producing anti-IL-17 autoantibodies.

[0109] According to an exemplified embodiment, the anti-IL-17 antibody is a human antibody directed against human interleukin IL-17 obtained after immortalization of human B lymphocytes derived from PBMC cells (“ Peripheral blood mononuclear cells”) of individuals with rheumatoid arthritis and producing anti-IL-17 autoantibodies.

[0110] Preferably, said anti-IL-17 antibodies are monoclonal antibodies.

[0111] The anti-IL-17 antibody may be a human antibody directed against human interleukin (IL17A), human IL-17F, or directed against both human IL-17A and human IL-17F.

[0112] According to a preferred embodiment, the anti-IL-17 antibody is an anti-IL-17A antibody.

[0113] Such an antibody may be identical or substantially identical to a natural antibody, for example produced from antibody-producing cells, such as B lymphocytes from individuals with rheumatoid arthritis, such as individuals with non-destructive rheumatoid arthritis and / or rheumatoid arthritis associated with the production of anti-IL-17 autoantibodies.

[0114] Thus, the anti-IL-17 autoantibody may preferably be a human autoantibody, directed against human interleukin IL-17A, human IL-17F, or directed against both human IL-17A and human IL-17F.

[0115] An isolated anti-IL-17 antibody as defined above can be implemented in a pharmaceutical composition and / or as a medicament.

[0116] The invention therefore also relates to a pharmaceutical composition or a medicament comprising said anti-IL-17 antibody.

[0117] The invention is defined by the appended claims. EXAMPLES MATERIALS AND METHODS

[0118] Patients: Sixty patients with rheumatoid arthritis (RA) were selected from a large database and classified into two groups in a 1:1 ratio based on their radiographic degree of destruction (Larsen score). Destructive rheumatoid arthritis (RA) is defined by a wrist Larsen score greater than 2, and non-destructive RA between 0 and 1. All RA-related parameters were obtained from the clinical database. Destructive and non-destructive RA patients were matched for sex (female / male = 21 / 09 vs. 20 / 10), age (66.1 ± 10.8 vs. 71.3 ± 9.3 years), disease duration (18.6 ± 9.6 vs. 23.0 ± 9.8 years), DAS28 (3.9 ± 1.2 vs. 4.0 ± 1.4), except for Larsen score (0.5 ± 0.5 vs. 3.3 ± 1.0, p < 0.0001) (Table 1). Thirty healthy donors were used as negative controls. Written consent was obtained from each subject. The protocol is in accordance with the Ethics Committee of the Lyon Hospitals.

[0119] Detection of anti-IL-17 antibodies by a competitive ELISA test ( Figure 1A ): Plasma from RA patients and healthy donors was first pre-incubated with horse serum overnight to prevent cross-reactions with rheumatoid factor (RF). Plasma samples at 1 / 4, 1 / 8, and 1 / 16 dilutions were then incubated with 30 µl of IL-17A (50 ng / ml) (IL17A, Dendritics, Lyon, France). After 1 h of incubation, an anti-human IL-17 detection antibody (406G9.02-HRP, Dendritics, Lyon, France) was added. This mixture was transferred to a 96-well plate containing a mouse anti-human IL-17 antibody (408H6.01, Dendritics, Lyon, France) for 2 h. Tetramethylbenzidine (TMB) substrate was added, and the absorbance at 620 nm was determined.

[0120] Detection of IL-17 complexes / anti-IL-17 antibodies by an indirect ELISA test ( Figure 1B ) : the wells are incubated overnight with 3 µg / mL of an anti-IL-17 capture antibody (408H6.01). After washing with PBS / Tween 0.05%, 100 µl of plasma diluted to ¼ are added overnight (first dilution to ½ in horse serum and second dilution in a PBS / BSA / Tween mixture). After 3 washes, a goat anti-human IgG Fc Fragment antibody (anti-human IgG Fc) conjugated to peroxidase (109-035-098, Jackson Immuno research, Baltimore, USA) at 1 / 5000 is added and incubated for 1h30. As a positive control, a mixture of a human anti-human IL-17A antibody isolated from the blood of a patient with rheumatoid arthritis (5µg / ml) and an interleukin IL-17 at 500 ng / ml was used.This human antibody against human interleukin IL-17 was obtained after immortalization of human B lymphocytes from PBMC cells of patients with rheumatoid arthritis and after immortalization by EBV and the CD40 system (DDXK-HuBBB, Dendritics, Lyon, France). To verify the elimination of rheumatoid factor, the reaction was tested with and without horse serum.

[0121] Statistical analysis: Data were expressed as mean ± SD. A non-parametric two-tailed t-test in Graphpad Prism software was used. A value p ( p value) less than 0.05 is considered statistically significant. EXAMPLE 1 - High incidence of anti-IL-17 autoantibodies in patients with non-destructive rheumatoid arthritis.

[0122] A competitive ELISA assay was developed to measure anti-IL17 autoantibodies in plasma. To prevent cross-reactivity with rheumatoid factor, which is present in large amounts in plasma, a pre-incubation step in the presence of horse serum was added. A positive control assay using purified anti-IL-17 antibodies showed a decrease in absorbance with dilution ( fig 1A ), while non-specific antibodies showed no variation.

[0123] Plasma from 30 healthy donors allowed the identification of a threshold value. An absorbance of 0.9 ± 0.1 was observed for a ½ dilution, and no variation for dilutions of ¼ to 1 / 8, which indicates an absence of positivity for anti-IL-17 autoantibodies.

[0124] Conversely, anti-IL-17 antibodies were detected in 36.6% of the 60 patients with rheumatoid arthritis ( p< 0.05 vs. controls), indicating a link between the presence of autoantibodies directed against interleukin IL-17 and rheumatoid arthritis.

[0125] To study the relationship between the severity of rheumatoid arthritis, plasma from individuals with destructive and non-destructive rheumatoid arthritis was collected. The different groups were matched, as indicated in the Materials & Methods section (see table 1 below). The so-called "non-destructive" patients had presented this pathology for several years, in order to confirm the reduced severity. Table 1: Clinical parameters of healthy donors, destructive and non-destructive rheumatoid arthritis, and incidence of anti-IL-17 autoantibodies Settings Healthy donors (n=30) PR patients (n=60) Non-destructive (n=30) Destroyer (n=30) p value Gender (F:M) 20:10 41 :19 21:09 20:10 ns Age (years) 60.0 ± 5.5 68.7 ± 10.5 66.1 ± 10.8 71.3 ± 9.3 ns Duration of illness (years) 20.8 ± 9.7 18.6 ± 9.6 23.0 ± 9.8 ns DAS28 4.0 ± 1.3 3.9 ± 1.2 4.0 ± 1.4 ns Larsen Score 1.9 ± 0.8 0.5 ± 0.5 3.3 ± 1.0 < 0.0001 Rheumatoid factor positive (%) 63.2 57.1 69.2 ns Anti-CCP antibody positive (%) 58.3 50.0 66.6 ns Anti-IL-17 antibodies (%) 0.0 35.4 46.6 24.2 < 0.05 ns = not significant, F = female, M = male, Abs = antibodies, DAS28 = disease activity score or Disease Activity Score 28.

[0126] Anti-IL-17 antibodies were detected in 46.6% of "non-destructive" patients, whereas these antibodies were detected in only 24.2% of "destructive" patients. These results suggest that natural anti-IL-17 autoantibodies are associated with rheumatoid arthritis with a better prognosis, regarding bone destruction.

[0127] Similar observations have been reported for other cytokines. In particular, increased levels of autoantibodies directed against IL-1α in patients with rheumatoid arthritis suggest a protective role of these anti-IL-1α antibodies against bone destruction.

[0128] Conversely, autoantibodies against IL-8 and osteopontin have been associated with extra-articular manifestations of rheumatoid arthritis.

Claims

1. In vitro method for evaluating the prognosis of a chronic autoimmune or inflammatory disease in an individual, comprising the following steps: a) determining (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from said individual, and b) comparing the level of autoantibody and / or of complex determined in step a) with a reference value, said comparison being indicative of the prognosis of a chronic autoimmune or inflammatory disease in said individual; in which the chronic autoimmune or inflammatory disease is rheumatoid arthritis (RA).

2. In vitro method according to Claim 1, for evaluating the prognosis of a chronic autoimmune or inflammatory disease in an individual for (i) determining the risk of bone destruction in said individual suffering from said chronic autoimmune or inflammatory disease; and / or (ii) determining the chances of response, of said individual suffering from said chronic autoimmune or inflammatory disease, to a treatment comprising the administration of an IL-17-inhibiting active ingredient; and / or (iii) determining the efficacy of a treatment or of a prevention of bone destruction in said individual suffering from said chronic autoimmune or inflammatory disease, said treatment or said prevention consisting of the administration of an IL-17-inhibiting active ingredient; in which the comparison in step b) is indicative of said risk of bone destruction, of said chances of response to said treatment, and / or of the efficacy of said treatment or of said prevention of bone destruction.

3. In vitro method for selecting an antibody for treating (i) a chronic autoimmune or inflammatory disease, and / or (ii) bone destruction associated with said disease, in which the chronic autoimmune or inflammatory disease is rheumatoid arthritis (RA), comprising the following steps: a) determining the level (i) of an anti-IL-17 autoantibody and / or (ii) of an [IL 17 / anti-IL-17 autoantibody] complex in a biological sample from an individual evaluated by the method according to Claim 1; b) selecting an anti-IL-17 autoantibody or a cell producing an anti-IL-17 autoantibody from a biological sample of said individual; said autoantibody being capable of treating said disease and / or said bone destruction.

4. In vitro method for distinguishing destructive rheumatoid arthritis (RA) from non-destructive RA in an individual, comprising the following steps: a) determining (i) the level of an anti-IL-17 autoantibody and / or (ii) the level of an [IL-17 / anti-IL-17 autoantibody] complex in a biological sample from said individual, the biological sample being chosen from whole blood, plasma and serum, and b) comparing the level of autoantibody and / or of complex determined in step a) with a reference value, said reference value being determined (i) in individuals who do not have bone destruction and / or whose rheumatoid arthritis is associated with the production of anti-IL-17 autoantibody and / or of [IL-17 / anti-IL-17 autoantibody] complex, or (ii) in individuals with bone destruction and / or in individuals whose rheumatoid arthritis is not associated with the production of anti-IL-17 autoantibody and / or of [IL-17 / anti-IL-17 autoantibody] complex, said comparison being indicative of destructive or non-destructive rheumatoid arthritis (RA) in said individual.

5. In vitro method according to one of the preceding claims, in which step a) consists in determining the level of an anti-IL-17 autoantibody in said biological sample.

6. In vitro method according to one of the preceding claims, in which step a) consists in determining the level of an [IL-17 / anti-IL-17 autoantibody] complex in said biological sample.

7. In vitro method according to one of Claims 1-3, in which the biological sample is chosen from: whole blood, plasma, serum, synovial fluid, cerebrospinal fluid, pleural fluid and peritoneal fluid.

8. In vitro method according to one of the preceding claims, in which the anti-IL-17 autoantibody is an anti-IL-17A autoantibody; and the [IL-17 / anti-IL-17 autoantibody] complex is an [IL-17A / anti-IL-17A autoantibody] complex.

9. In vitro method according to one of the preceding claims, in which step a) is carried out in the form of an ELISA assay.

10. In vitro method according to one of the preceding claims, also comprising a step of determining, in a biological sample from said individual, the level of a polypeptide chosen from: the cytokines IL-17A, IL-17F, IL-25, IL-23, the transcription factor RORγt, and the IL-17RA, IL-17RB, IL-17RC receptors, autoantibodies directed against IL-1α, autoantibodies directed against IL-8 and / or autoantibodies directed against osteopontin.

11. Isolated anti-IL-17 antibody, for use in the treatment or prevention (i) of destructive rheumatoid arthritis or (ii) of rheumatoid arthritis in an individual not producing anti-IL-17 autoantibodies, or of bone destruction associated with either one of groups (i) or (ii), said individual being identified by the method according to one of Claims 4 to 10.