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Multimers of MHC-Peptide Complexes and Uses Thereof in Borrelia Infectious Diseases

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Multimers of MHC-Peptide Complexes and Uses Thereof in Borrelia Infectious Diseases (19) TZZ ¥_¥Z T (11) EP 2 361 930 A2 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) Int Cl.: 31.08.2011 Bulletin 2011/35 C07K 14/705 (2006.01) A61K 47/48 (2006.01) A61K 39/00 (2006.01) A61K 38/17 (2006.01) (2006.01) (2006.01) (21) Application number: 11150404.9 G01N 33/50 A61P 31/00 A61K 39/02 (2006.01) (22) Date of filing: 26.03.2008 (84) Designated Contracting States: (72) Inventors: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR • Schøller, Jørgen HR HU IE IS IT LI LT LU LV MC MT NL NO PL PT 2800 Lyngby (DK) RO SE SI SK TR • Pedersen, Henrik 3540 Lynge (DK) (30) Priority: 26.03.2007 DK 200700461 • Brix, Liselotte 26.03.2007 US 907217 P 2880 Bagsværd (DK) 03.07.2007 DK 200700973 03.07.2007 DK 200700975 (74) Representative: Aamand, Jesper L. et al 03.07.2007 DK 200700972 Hjerrild & Levin ApS 03.07.2007 DK 200700974 Tuborg Boulevard 12 03.07.2007 US 929583 P 2900 Hellerup (DK) 03.07.2007 US 929581 P 03.07.2007 US 929582 P Remarks: 03.07.2007 US 929586 P •ThecompletedocumentincludingReferenceTables and the Sequence Listing can be downloaded from (62) Document number(s) of the earlier application(s) in the EPO website accordance with Art. 76 EPC: •This application was filed on 07-01-2011 as a 08715595.8 / 2 155 782 divisional application to the application mentioned under INID code 62. (71) Applicant: Dako Denmark A/S 2600 Glostrup (DK) (54) Multimers of MHC-peptide complexes and uses thereof in Borrelia infectious diseases (57) The present application concerns multimers of MHC-peptide complexes, in particular, dextramers, wherein the peptide is a 9-mer derived from a Borrelia outer surface protein antigen. The application is also di- rected to compositions, kits and vaccines comprising said MHC multimers and uses thereof in immuno-moni- toring, diagnosis, and therapy, especially to detect spe- cific T cells. EP 2 361 930 A2 Printed by Jouve, 75001 PARIS (FR) EP 2 361 930 A2 Description [0001] All patent and non-patent references cited in US 60/907,217 as well as in this application are hereby incorporated by reference in their entirety. US 60/907,217 is hereby also incorporated herein by reference in its entirety. 5 Field of invention [0002] The present invention relates to MHC-peptide complexes and uses thereof in the treatment of a disease in an individual. 10 Background of invention [0003] Biochemical interactions between peptide epitope specific membrane molecules encoded by the Major Histo- compatibility Complex (MHC, in humans HLA) and T-cell receptors (TCR) are required to elicit specific immune responses. 15 This requires activation of T-cells by presentation to the T-cells of peptides against which a T-cell response should be raised. The peptides are presented to the T-cells by the MHC complexes. The immune response 20 [0004] The immune response is divided into two parts termed the innate immune response and the adaptive immune response. Both responses work together to eliminate pathogens (antigens). Innate immunity is present at all times and is the first line of defense against invading pathogens. The immediate response by means of pre-existing elements, i.e. various proteins and phagocytic cells that recognize conserved features on the pathogens, is important in clearing and control of spreading of pathogens. If a pathogen is persistent in the body and thus only partially cleared by the actions 25 of the innate immune system, the adaptive immune system initiate a response against the pathogen. The adaptive immune system is capable of eliciting a response against virtually any type of pathogen and is unlike the innate immune system capable of establishing immunological memory. [0005] The adaptive response is highly specific to the particular pathogen that activated it but it is not so quickly launched as the innate when first encountering a pathogen. However, due to the generation of memory cells, a fast and 30 more efficient response is generated upon repeated exposure to the same pathogen. The adaptive response is carried out by two distinct sets of lymphocytes, the B cells producing antibodies leading to the humoral or antibody mediated immune response, and the T cells leading to the cell mediated immune response. [0006] T cells express a clonotypic T cell receptor (TCR) on the surface. This receptor enable the T cell to recognize peptide antigens bound to major histocompatibility complex (MHC) molecules, called human leukocyte antigens (HLA) 35 in man. Depending on the type of pathogen, being intracellular or extracellular, the antigenic peptides are bound to MHC class I or MHC class II, respectively. The two classes of MHC complexes are recognized by different subsets of T cells; Cytotoxic CD8+ T cells recognizing MHC class I and CD4+ helper cells recognizing MHC class II. In general, TCR recognition of MHC-peptide complexes result in T cell activation, clonal expansion and differentiation of the T cells into effector, memory and regulatory T cells. 40 [0007] B cells express a membrane bound form of immunoglobulin (Ig) called the B cell receptor (BCR). The BCR recognizes an epitope that is part of an intact three dimensional antigenic molecule. Upon BCR recognition of an antigen the BCR:antigen complex is internalized and fragments from the internalized antigen is presented in the context of MHC class II on the surface of the B cell to CD4+ helper T-cells (Th). The specific Th cell will then activate the B cell leading to differentiation into an antibody producing plasma cell. 45 [0008] A very important feature of the adaptive immune system is its ability to distinguish between self and non-self antigens, and preferably respond against non-self. If the immune system fails to discriminate between the two, specific immune responses against self-antigens are generated. These autoimmune reactions can lead to damage of self-tissue. [0009] The adaptive immune response is initiated when antigens are taken up by professional antigen presenting cells such as dendritic cells, Macrophages, Langerhans cells and B-cells. These cells present peptide fragments, resulting 50 from the degradation of proteins, in the context of MHC class II proteins (Major Histocompatibility Complex) to helper T cells. The T helper cells then mediate help to B-cells and antigen specific cytotoxic T cells, both of which have received primary activation signals via their BCR respective TCR. The help from the Th-cell is mediated by means of soluble mediators e.g. cytokines. [0010] In general the interactions between the various cells of the cellular immune response is governed by receptor- 55 ligand interactions directly between the cells and by production of various soluble reporter substances e.g. cytokines by activated cells. 2 EP 2 361 930 A2 MHC-peptide complexes. [0011] MHC complexes function as antigenic peptide receptors, collecting peptides inside the cell and transporting them to the cell surface, where the MHC-peptide complex can be recognized by T-lymphocytes. Two classes of classical 5 MHC complexes exist, MHC class I and II. The most important difference between these two molecules lies in the protein source from which they obtain their associated peptides. MHC class I molecules present peptides derived from endog- enous antigens degraded in the cytosol and are thus able to display fragments of viral proteins and unique proteins derived from cancerous cells. Almost all nucleated cells express MHC class I on their surface even though the expression level varies among different cell types. MHC class II molecules bind peptides derived from exogenous antigens. Exog- 10 enous proteins enter the cells by endocytosis or phagocytosis, and these proteins are degraded by proteases in acidified intracellular vesicles before presentation by MHC class II molecules. MHC class II molecules are only expressed on professional antigen presenting cells like B cells and macrophages. [0012] The three-dimensional structure of MHC class I and II molecules are very similar but important differences exist. MHC class I molecules consist of two polypeptide chains, a heavy chain, α, spanning the membrane and a light 15 chain, β2-microglobulin (β2m). The heavy chain is encoded in the gene complex termed the major histocompatibility complex (MHC), and its extracellular portion comprises three domains, α1, α2 and α3. The β2m chain is not encoded in the MHC gene and consists of a single domain, which together with the α3 domain of the heavy chain make up a folded structure that closely resembles that of the immunoglobulin. The α1 and α2 domains pair to form the peptide binding cleft, consisting of two segmented α helices lying on a sheet of eight β-strands. In humans as well as in mice 20 three different types of MHC class I molecule exist. HLA-A, B, C are found in humans while MHC class I molecules in mice are designated H-2K, H-2D and H-2L. [0013] The MHC class II molecule is composed of two membrane spanning polypeptide chains, α and β, of similar size (about 30000 Da). Genes located in the major histocompatibility complex encode both chains. Each chain consists of two domains, where α1 and β1 forms a 9-pocket peptide-binding cleft, where pocket 1, 4, 6 and 9 are considered as 25 major peptide binding pockets. The α2 and β2, like the α2 and β2m in the MHC class I molecules, have amino acid sequence and structural similarities to immunoglobulin constant domains. In contrast to MHC class I complexes, where the ends of the antigenic peptide is buried, peptide-ends in MHC class II complexes are not.
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