MHC Class II molecules and their usage

Abstract

The present disclosure relates to HLA class II molecules having higher affinity for CD4 than naturally occurring HLA class II molecules. In certain aspects, the HLA class II molecules comprise a DQ beta chain having: (i) an amino acid other than leucine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1, (ii) an amino acid other than valine at a position corresponding to amino acid residue 143 of SEQ ID NO: 1, or (iii) both (i) and (ii). Certain aspects of the present disclosure relate to nucleic acid molecules encoding the HLA class II molecules, vectors comprising the nucleic acid molecules, cells comprising them, and methods of using the same.

Description

[0001] Cross-reference to related applications

[0002] This PCT application claims priority to U.S. Provisional Application No. 62 / 880,501, filed July 30, 2019, and U.S. Provisional Application No. 63 / 029,114, filed May 22, 2020, each of which is incorporated herein by reference in its entirety.

[0003] Through EFS-WEB Electronic

[0004] References to the submitted sequence list

[0005] The contents of the sequence list submitted electronically (name: 4285-011PC02_SL_ST25.txt, size: 34,403 bytes; creation date: July 28, 2020) are incorporated herein by reference in their entirety. Invention Field

[0006] This disclosure provides major histocompatibility complex (MHC) class II molecules with increased affinity for CD4 and their uses. Background of the Invention

[0008] Immunotherapy has become a key tool in the fight against a variety of diseases, including cancer. T-cell therapy is at the forefront of immunotherapy development, and adoptive transfer of anti-tumor T cells has been shown to induce clinical responses in cancer patients.

[0009] Targeted T-cell therapy, using T-cell receptors (TCRs) expressed by T cells that specifically target epitopes expressed by tumor cells, is a promising form of T-cell therapy. Antigen-presenting cells display peptide fragments associated with the major histocompatibility complex (MHC) on their surface to induce an immune response. Improved presentation of endogenous peptides via class II has been shown to be associated with improved survival in cancer patients. However, the development of novel TCRs capable of specifically targeting MHC class II presenting peptides is hampered by the low affinity of MHC class II proteins for CD4 expressed by T cells.

[0010] This disclosure provides MHC class II proteins with increased affinity for CD4 and methods for using them to identify and develop novel MHC class II specific TCRs. Summary of the Invention

[0011] Some aspects of this disclosure relate to HLA class II molecules comprising a DQβ chain, wherein the DQβ chain comprises an amino acid other than leucine at the position of amino acid residue 114 corresponding to SEQ ID NO:1.

[0012] Certain aspects of this disclosure relate to HLA class II molecules comprising a DQβ chain, wherein the DQβ chain contains a substitution mutation at the position of amino acid residue 114 corresponding to SEQ ID NO:1, wherein the substitution mutation is a substitution of an amino acid other than leucine.

[0013] In some respects, the DQβ chain also contains amino acids other than valine at the position of amino acid residue 143 corresponding to SEQ ID NO:1.

[0014] Some aspects of this disclosure relate to HLA class II molecules comprising a DQβ chain, wherein the DQβ chain comprises an amino acid other than valine at the position of amino acid residue 143 corresponding to SEQ ID NO:1.

[0015] Certain aspects of this disclosure relate to HLA class II molecules comprising a DQβ chain, wherein the DQβ chain contains a substitution mutation at the position of amino acid residue 143 corresponding to SEQ ID NO:1, wherein the substitution mutation is a substitution of an amino acid other than valine.

[0016] In some aspects, the DQβ chain further includes an amino acid other than leucine at the position corresponding to amino acid residue 114 of SEQ ID NO:1. In some aspects, the DQβ chain further includes an amino acid other than asparagine at the position corresponding to amino acid residue 110 of SEQ ID NO:1. In some aspects, the DQβ chain further includes an amino acid other than isoleucine at the position corresponding to amino acid residue 116 of SEQ ID NO:1. In some aspects, the DQβ chain further includes an amino acid other than serine at the position corresponding to amino acid residue 118 of SEQ ID NO:1. In some aspects, the DQβ chain further includes an amino acid other than proline at the position corresponding to amino acid residue 146 of SEQ ID NO:1.

[0017] In some respects, the DQβ chain also comprises at least three of the following: (i) an amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1, (ii) an amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1, (iii) an amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1, and (iv) an amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1.

[0018] In some respects, the DQβ chain also comprises: (i) an amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1, (ii) an amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1, (iii) an amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1, and (iv) an amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1.

[0019] In some aspects, the amino acid at position 114 of amino acid residue SEQ ID NO:1, excluding leucine, comprises a hydrophobic side chain. In some aspects, the amino acid at position 114 of amino acid residue SEQ ID NO:1, excluding leucine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, the amino acid at position 114 of amino acid residue SEQ ID NO:1, excluding leucine, is tryptophan.

[0020] In some aspects, the amino acid at position 143 of amino acid residue SEQ ID NO:1, excluding valine, comprises a hydrophobic side chain. In some aspects, the amino acid at position 143 of amino acid residue SEQ ID NO:1, excluding valine, is selected from alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, the amino acid at position 143 of amino acid residue SEQ ID NO:1, excluding valine, is methionine.

[0021] In some aspects, the β-chain of an MHC class II molecule contains an amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1 is selected from serine, threonine, and glutamine. In some aspects, the amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1 is glutamine.

[0022] In some aspects, the β-chain of an MHC class II molecule contains an amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1 is selected from alanine, valine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, the amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1 is valine.

[0023] In some aspects, the β-chain of an MHC class II molecule contains an amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1 is selected from arginine, histidine, and lysine. In some aspects, the amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1 is histidine.

[0024] In some aspects, the β-chain of an MHC class II molecule contains an amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1 is selected from serine, threonine, asparagine, and glutamine. In some aspects, the amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1 is glutamine.

[0025] In some respects, the DQβ chain contains an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with an amino acid sequence selected from SEQ ID NO:1, 3, 4, and 5.

[0026] In some respects, the DQβ chain contains tryptophan at position 114 of amino acid residue SEQ ID NO:1 and methionine at position 143 of amino acid residue SEQ ID NO:1.

[0027] In some respects, the β chain of an MHC class II molecule comprises (a) tryptophan at position 114 of amino acid residue SEQ ID NO:1; (b) methionine at position 143 of amino acid residue SEQ ID NO:1; and (c) at least one of the following: (i) glutamine at position 110 of amino acid residue SEQ ID NO:1; (ii) valine at position 116 of amino acid residue SEQ ID NO:1; (iii) histidine at position 118 of amino acid residue SEQ ID NO:1; and (iv) glutamine at position 146 of amino acid residue SEQ ID NO:1.

[0028] In some respects, the β chain of an MHC class II molecule comprises (a) tryptophan at position 114 of amino acid residue SEQ ID NO:1; (b) methionine at position 143 of amino acid residue SEQ ID NO:1; and (c) at least two of the following: (i) glutamine at position 110 of amino acid residue SEQ ID NO:1; (ii) valine at position 116 of amino acid residue SEQ ID NO:1; (iii) histidine at position 118 of amino acid residue SEQ ID NO:1; and (iv) glutamine at position 146 of amino acid residue SEQ ID NO:1.

[0029] In some respects, the β chain of an MHC class II molecule comprises (a) tryptophan at position 114 of amino acid residue SEQ ID NO:1; (b) methionine at position 143 of amino acid residue SEQ ID NO:1; and (c) at least three of the following: (i) glutamine at position 110 of amino acid residue SEQ ID NO:1; (ii) valine at position 116 of amino acid residue SEQ ID NO:1; (iii) histidine at position 118 of amino acid residue SEQ ID NO:1; and (iv) glutamine at position 146 of amino acid residue SEQ ID NO:1.

[0030] In some respects, the β chain of an MHC class II molecule contains (a) tryptophan at position 114 of amino acid residue SEQ ID NO:1; (b) methionine at position 143 of amino acid residue SEQ ID NO:1; (c) glutamine at position 110 of amino acid residue SEQ ID NO:1; (d) valine at position 116 of amino acid residue SEQ ID NO:1; (e) histidine at position 118 of amino acid residue SEQ ID NO:1; and (f) glutamine at position 146 of amino acid residue SEQ ID NO:1.

[0031] In some aspects, the DQβ chain contains the amino acid sequence shown in SEQ ID NO:3. In some aspects, the DQβ chain contains the amino acid sequence shown in SEQ ID NO:4.

[0032] In some aspects, the β chain of HLA class II molecules contains alleles of DQ2, DQ3, DQ4, DQ5, or DQ6. In other aspects, the β chain of MHC class II molecules contains alleles of HLA-DQB1*02, HLA-DQB1*03, HLA-DQB1*04, HLA-DQB1*05, or HLA-DQB1*06.

[0033] In some respects, HLA class II molecules also contain the DQα chain. In some respects, the α chain of MHC class II molecules contains the HLA-DQA1*01, HLA-DQA1*02, HLA-DQA1*03, HLA-DQA1*04, HLA-DQA1*05, or HLA-DQA1*06 alleles.

[0034] In some aspects, the DQα chain comprises an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with SEQ ID NO:6 or 8. In some aspects, the DQα chain comprises the amino acid sequence shown in SEQ ID NO:6 or 8.

[0035] In some respects, the DQβ chain has increased affinity for CD4 protein compared to a reference HLA class II molecule, wherein the DQβ chain contained in the reference HLA class II molecule contains (i) leucine at position 114 of amino acid residue SEQ ID NO:1 and / or (ii) valine at position 143 of amino acid residue SEQ ID NO:1.

[0036] In some respects, the increased affinity is at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 15 times, at least about 20 times, at least about 25 times, at least about 30 times, at least about 35 times, at least about 40 times, at least about 45 times, at least about 50 times, at least about 75 times, at least about 100 times, at least about 200 times, at least about 300 times, at least about 400 times, at least about 500 times, or at least about 1000 times.

[0037] In some respects, the DQβ chain binds to the cell membrane. In other respects, the DQβ chain does not bind to the cell membrane. In some respects, the DQβ chain contains the extracellular domain of the full-length DQα chain. In other respects, the DQβ chain does not contain the transmembrane domain of the full-length DQβ chain.

[0038] In some respects, the DQα chain binds to the cell membrane. In other respects, the DQα chain does not bind to the cell membrane. In some respects, the DQα chain contains the extracellular domain of the full-length DQα chain. In other respects, the DQα chain does not contain the transmembrane domain of the full-length DQα chain.

[0039] In some respects, the DQβ chains are linked or associated with inert particles. In some respects, the inert particles are beads. In some respects, the inert particles are nanoparticles. In some respects, the nanoparticles are selected from polyethylene glycolated iron oxide, chitosan, dextran, gelatin, alginate, liposomes, starch, branched polymers, carbon-based carriers, polylactic acid, poly(cyano)acrylate, polyethyleneimine, block copolymers, polycaprolactone, SPIONs, USPIONs, Cd / Zn-selenide, or silica nanoparticles. In some respects, the nanoparticles are polyethylene glycolated iron oxide nanoparticles.

[0040] In some respects, the DQβ chain contains a signal peptide. In some respects, the DQα chain contains a signal peptide. In some respects, the signal peptide contains the amino acid sequence shown in SEQ ID NO:9.

[0041] Some aspects of this disclosure relate to nucleic acid molecules encoding the DQβ chain disclosed herein. In some aspects, said nucleic acid molecules further encode the DQα chain disclosed herein.

[0042] In some respects, the nucleic acid molecule comprises a nucleotide sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with SEQ ID NO:2.

[0043] Certain aspects of this disclosure relate to vectors containing nucleic acid molecules disclosed herein.

[0044] Certain aspects of this disclosure relate to treating cells comprising the HLA class II molecules, nucleic acid molecules, or vectors disclosed herein. In some aspects, the cells are mammalian or insect cells. In some aspects, the cells are selected from K562 cells, T2, HEK293, HEK293T, A375, SK-MEL-28, Me275, COS, fibroblasts, tumor cells, or any combination thereof. In some aspects, the cells lack endogenous MHC class II DQβ chain expression. In some aspects, the cells lack endogenous MHC class II DQα chain expression.

[0045] Certain aspects of this disclosure relate to methods for identifying T-cell receptors capable of binding to epitopes in MHC class II complexes, said methods comprising shocking cells disclosed herein with one or more peptides containing said epitopes and stimulating one or more CD4+ cells with APCs. + T cells.

[0046] Certain aspects of this disclosure relate to methods for treating a disease or condition in a subject in need, said methods comprising administering to the subject an MHC class II molecule disclosed herein. In some aspects, the disease or condition is cancer or an infection.

[0047] In some respects, cancer is selected from the group consisting of: melanoma, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, rectal cancer, stomach cancer, uterine cancer, lung cancer, Hodgkin's disease, non-Hodgkin's lymphoma (NHL), esophageal cancer, small bowel cancer, urethral cancer, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia (ALL) (including non-T-cell ALL), chronic lymphocytic leukemia (CLL), bladder cancer, kidney or ureteral cancer, renal pelvis cancer, glioma, squamous cell carcinoma, and combinations thereof.

[0048] In some respects, cancer is recurrent or refractory. In some respects, cancer is locally advanced. In some respects, cancer is advanced. In some respects, cancer is metastatic.

[0049] In some respects, HLA class II molecules have a K+ level of less than approximately 100 μM. D Binding to CD4. In some respects, HLA class II molecules bind with K at concentrations less than approximately 10 μM. D Binding to CD4. In some respects, HLA class II molecules bind at approximately 8.9 μM or less K. D Combined with CD4.

[0050] Certain aspects of this disclosure relate to complexes comprising the HLA class II molecules and peptides disclosed herein, wherein said peptides comprise DDX3Y.171-190 HA 255-270 GPC3 138-157 Or any combination thereof. Attached Figure Description

[0051] Figures 1A to 1F Data are provided illustrating the enhanced CD4 binding ability of the modified DQ molecule. Figure 1A It compares DPB1*04:01, DQB1*05:01, and DQB1*05:01. L114W / V143M+4reps A table of amino acid sequences, with mutated amino acids underlined. Figure 1B and Figure 1C Stable expression of wild-type DQ5 (DQA1*01:01 / DQB1*05:01) using sCD4 staining. L114W / V143M DQ5 L114W / V143M+4reps Wild-type DP4 or DP4 L112W / V141M (like Figure 1A A graphical representation of data for type II defective K562 cells (shown). Figure 1D This shows DQ5 cells similarly stained with sCD4, each expressing a single amino acid inversion at one of the four positions. L114W / V143M+4reps CD4 binding ability of a series of K562 derivatives of the mutant. Figure 1E This is a table listing the amino acid sequences of DPB1*04:01, DQB1*02:01, DQB1*04:02, and DQB1*06:01, with substituted amino acids underlined. Note that, unlike DQB1*05:01, DQB1*02:01, DQB1*04:02, and DQB1*06:01 encode Val at position 116, similar to DPB1*04:01, which encodes Val at position 114. Figure 1F Graphical representations are provided showing the data on L114W / V143M+3reps substitution enhancements of DQ2, DQ4, and DQ6 binding to CD4 in the β chain. At least two independent experiments were conducted. *, P < 0.05, obtained via Student's t-test. Bars and error bars represent the mean ± SD of triplicate experimental results.

[0052] Figures 2A to 2B This is a graphical representation illustrating the detection of affinity-matured DQ dimers in homologous TCRs expressed in human primary CD4+ T cells. It shows the DQ5(DQA1*01:01-DQB1*05:01)-restricted DDX3Y-specific TCR (E6) (…). Figure 2A ) and DQ6 (DQA1*01:02-DQB1*06:02) restricted influenza virus HA-specific TCR (DM2) ( Figure 2B Reconstructed in primary human CD4+ T cells, and administered via DQ5.L114W / V143M+4reps and DQ6 L114W / V143M+3reps Dimer staining. Perform at least two independent experiments.

[0053] Figures 3A to 3Q This is a graphical representation of the equivalent expression levels of HLA class II genes using histograms. HLA-DQ and its derivatives were reconstructed in K562 cells and stained with anti-HLA class II monoclonal antibodies. Anti-HLA class II monoclonal antibody clone 9-49(I3) (DQ5 and DQ6) or anti-class II monoclonal antibody clone... (DQ2 and DQ4) Surface expression of each DQ2, DQ5, and DQ6 allele was detected. Open histograms represent isotype control staining.

[0054] Figures 4A to 4L It displays DQ5 L114W / V143M+4reps Dimer robust staining E6-transduced CD4 + Graphical representation of T cells. E6 in CD4 + Reconstruction in T cells, then using wild-type DQ5 ( Figure 4D and Figure 4J DQ5 L114W / V143M ( Figure 4E and Figure 4K ) and DQ5 L114W / V143M+4reps ( Figure 4F and Figure 4L CLIP control dimer ( Figures 4D to 4F ) and DDX3Y 171-190 Specific dimers ( Figures 4J to 4L ) staining. Control cells not transduced with TCR are shown in Figures 4A to 4C and Figures 4G to 4I middle.

[0055] Figures 5A to 5G This is a graphical representation showing the restriction TCR of the affinity-matured dimeric clone DQ5. From DQ5.1 + Purified primary CD4 from melanoma patients + T cells, and with irradiated GPC3 138-157 The expression of DQ5.1 was stimulated by aAPC. Two weeks later, homologous GPC3 was used. 138-157 -DQ5 L114W / V143M+4reps Dimer response to CD4 stimulation + T cells were stained ( Figures 5A to 5B GPC3-specific TCRs were reconstructed in TCR-deficient Jurkat76 / CD4 cells and analyzed via the corresponding DQ5 assay. L114W / V143M+4reps dimer ( Figure 5C (E6 / Control); Figure 5D (E6 / GPC3 138-157 ); Figure 5E (DQ5-06-GPC3 138-157 (comparison); and Figure 5F (DQ5-06-GPC3 138-157 / GPC3 138-157 )) staining. In the IL-2ELISPOT assay, Jurkat 76 / CD4 cells expressing GPC3-specific TCRs were stimulated with DQ5-K562 cells pulsed with the corresponding peptide ( Figure 5G ). Detailed Implementation

[0056] This disclosure relates to MHC class II molecules that have an increased affinity for CD4. In some aspects, this disclosure relates to MHC class II molecules comprising an HLA-DQ (DQ)β chain, wherein the DQβ chain has an increased affinity for CD4.

[0057] This disclosure further relates to MHC class II molecules comprising a DQβ chain, wherein the DQβ chain comprises an amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1. In some aspects, the DQβ chain also comprises an amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1.

[0058] This disclosure further relates to MHC class II molecules comprising a DQβ chain, wherein the DQβ chain comprises an amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1. In some aspects, the DQβ chain also comprises an amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1.

[0059] In some respects, the DQβ chain also comprises at least three of the following: (i) an amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1, (ii) an amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1, (iii) an amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1, and (iv) an amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1.

[0060] I. Terminology

[0061] To make this disclosure more readily understandable, certain terms are first defined. As used in this application, each of the following terms shall have the meaning described below unless expressly specified herein. Further definitions are set forth throughout the application.

[0062] It should be noted that the term "a / an" refers to one or more of the entities described; for example, "a nucleotide sequence" should be understood to mean one or more nucleotide sequences. Therefore, the terms "a," "one or more," and "at least one" are used interchangeably herein.

[0063] Furthermore, when used herein, “and / or” should be considered as each of the two specified features or components disclosed herein, with or without the other. Therefore, the term “and / or” as used in phrases herein, such as “A and / or B”, means to include: “A and B”; “A or B”; “A” (alone); and “B” (alone). Similarly, the term “and / or” as used in phrases, such as “A, B and / or C”, is intended to cover each of the following: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0064] The term "about" is used in this document to mean approximately, roughly, or around. When used with a numerical range, it modifies the range by extending the boundaries above and below the value. Generally, the term "about" is used in this document to modify a value by a variation of about 10% above or below the value (increase or decrease).

[0065] It should be understood that whenever an aspect is described in this document as “comprising”, other similar aspects described as “consisting of” and / or “substantially consisting of” are also provided.

[0066] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art relating to this invention. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd Edition, 2002, CRC Press; the Dictionary of Cell and Molecular Biology, 3rd Edition, 1999, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, Revised Edition, 2000, Oxford University Press provide a general dictionary for those skilled in the art of the use of many terms used in this disclosure.

[0067] Units, prefixes, and symbols are represented in their form acceptable to the Système International de Unites (SI). Numerical ranges include the numerical values ​​defining the range. Unless otherwise indicated, nucleotide sequences are written from left to right with a 5' to 3' orientation. Amino acid sequences are written from left to right with an amino to carboxyl orientation. The headings provided herein are not intended to limit the various aspects of this disclosure, which are derived from the entire specification. Therefore, the terms defined below are more fully defined by reference to the entire specification.

[0068] "Administration" means the physical introduction of an agent into a subject using any of the various methods and delivery systems known to those skilled in the art. Exemplary routes of administration for the formulations disclosed herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal, or other parenteral administration routes, such as by injection or infusion. As used herein, the phrase "parenteral administration" means a method of administration other than enteral and local administration, typically by injection, and including, but not limited to, intravenous, intramuscular, intraarterial, intrasheath, intralymphatic, intralesional, intracapsular, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as intracorporeal electroporation. In some aspects, the formulation is administered via non-parenteral routes, such as oral administration. Other non-parenteral routes include local, epidermal, or mucosal administration routes, such as intranasal, vaginal, rectal, sublingual, or local administration. Administration may also be performed, for example, once, multiple times, and / or over one or more extended periods of time.

[0069] As used herein, the term "HLA" refers to human leukocyte antigens. In humans, HLA genes encode major histocompatibility complex (MHC) proteins. MHC proteins are expressed on the surface of cells and are involved in the activation of immune responses. HLA class II genes encode MHC class II proteins expressed on the surface of professional antigen-presenting cells (APCs). Non-restricted examples of professional APCs include monocytes, macrophages, dendritic cells (DCs), and B lymphocytes. Some endothelial and epithelial cells also express MHC class II molecules after inflammatory signals are activated. Individuals lacking functional MHC class II molecules are highly susceptible to a range of infectious diseases and typically die at a young age.

[0070] As used herein, "HLA class II molecule" or "MHC class II molecule" refers to the protein product of a wild-type or variant HLA class II gene encoding an MHC class II molecule. Therefore, "HLA class II molecule" and "MHC class II molecule" are used interchangeably in this document. A typical MHC class II molecule contains two protein chains: an α chain and a β chain. Generally, naturally occurring α and β chains each contain a transmembrane domain that anchors the α / β chain to the cell surface, and an extracellular domain that carries antigens and interacts with the TCR and / or CD4 expressed on T cells.

[0071] Both the MHC class II α and β chains are encoded by the HLA gene complex. The HLA complex is located in the 6p21.3 region on the short arm of human chromosome 6 and contains more than 220 genes with different functions. It is known in the art that the HLA gene complex is highly variable, with more than 20,000 HLA alleles and associated alleles, including more than 250 MHC class II α chain alleles and 5,000 MHC class II β chain alleles, encoding thousands of MHC class II proteins (see, e.g., hla.alleles.org, last accessed May 20, 2019, incorporated herein by reference in its entirety). For example, one such HLA-DP allele, DP4, is the most common allele in many ethnic groups. The α and β chains are typically expressed as protoproteins, which also contain cleaved signal peptides. Any number of naturally occurring signal peptides can be used to facilitate the expression and localization of the α and β chains disclosed herein. One such example is SEQ ID NO:9.

[0072] Three loci in the HLA complex encode MHC class II proteins: HLA-DP, HLA-DQ, and HLA-DR. HLA-DO and HLA-DM encode proteins that associate with MHC class II molecules and support their conformation and function. Representative HLA-DQ sequences are provided in Table 1.

[0073] Table 1: Amino acid and nucleotide sequences of the DQ β and α chains.

[0074]

[0075]

[0076]

[0077] When MHC class II molecules complex with antigenic peptides, the 10-30 amino acid-long antigenic peptide binds to a peptide-binding groove and is presented extracellularly to CD4+ cells. Both the α and β chains fold into two separate domains: α-1 and α-2 of the α peptide, and β-1 and β-2 of the β peptide. The invariant residues at L114, V116, V143, L158, and M160, recognized and bound by CD4, are located in the β-2 domain of the β peptide. An open peptide-binding groove maintaining antigen presentation is found between the α-1 and β-1 domains. Upon interaction with CD4+ T cells, the MHC class II complex interacts with the T cell receptor (TCR) expressed on the T cell surface. Furthermore, the β chain of the MHC class II molecule interacts weakly with CD4+ receptors expressed on the T cell surface (K-R). D >2mM). The exemplary CD4 amino acid sequence (UniProt-P01730) is provided in Table 2 (SEQ ID NO:10).

[0078] Table 2: Human CD4 amino acid sequence

[0079]

[0080] As used herein, the term “T-cell receptor” (TCR) refers to a heteromeric cell surface receptor capable of specifically interacting with a target antigen. As used herein, “TCR” includes, but is not limited to, naturally occurring and non-natural TCRs, full-length TCRs and their antigen-binding portions, chimeric TCRs, TCR fusion constructs, and synthetic TCRs. In humans, TCRs are expressed on the surface of T cells, and they are responsible for T cell recognition and targeting by antigen-presenting cells. Antigen-presenting cells (APCs) display foreign protein (antigen) fragments that are complexed with major histocompatibility complexes (MHC class I or MHC class II) (also referred to herein as complexed with HLA molecules, such as HLA class II molecules). TCRs recognize and bind to the peptide:HLA complex and recruit CD8 (for MHC class I molecules) or CD4 (for MHC class II molecules) expressed by T cells, thereby activating the TCR. Activated TCRs initiate downstream signaling and immune responses, including the destruction of APCs.

[0081] Typically, a TCR can consist of two chains, an α-chain and a β-chain (or less commonly, a γ-chain and a δ-chain), interconnected by disulfide bonds. Each chain contains variable regions (α-chain variable domains and β-chain variable domains) and constant regions (α-chain constant regions and β-chain constant regions). The variable domains are located distal to the cell membrane and interact with antigens. The constant regions are located proximal to the cell membrane. A TCR may also contain a transmembrane region and a short cytoplasmic tail. As used herein, the term "constant region" includes the transmembrane region and the cytoplasmic tail, as well as (where present) the conventional "constant region".

[0082] The variable domains can be further subdivided into hypervariable regions called complementarity-determining regions (CDRs), which contain more conserved regions called framework regions (FRs). Each α-chain and β-chain variable domain contains three CDRs and four FRs: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. Each variable domain contains a binding domain that interacts with the antigen. Although all three CDRs on each chain are involved in antigen binding, CDR3 is considered the primary antigen-binding region, while CDR1 and CDR2 are considered to primarily recognize HLA molecules.

[0083] Unless explicitly stated otherwise, the term "TCR" also includes antigen-binding fragments or portions of any TCR disclosed herein, and includes monovalent and bivalent fragments or portions, as well as single-chain TCRs. The term "TCR" is not limited to naturally occurring TCRs that bind to the surface of T cells. As used herein, the term "TCR" further refers to TCRs expressed on the surface of cells other than T cells (e.g., cells naturally expressing or modified to express CD4 as described herein) or TCRs that do not have a cell membrane (e.g., isolated or soluble TCRs) as described herein.

[0084] "Antigen-binding molecule," "part of TCR," or "TCR fragment" refers to any portion of the TCR smaller than the whole. Antigen-binding molecules may include antigenic CDRs.

[0085] “Antigen” refers to any molecule, such as a peptide, that elicits an immune response or is capable of being bound by a TCR. As used herein, “epitope” refers to a portion of a polypeptide that elicits an immune response or is capable of being bound by a TCR. An immune response may involve the production of antibodies or the activation of specific immune-active cells, or both. Those skilled in the art will readily understand that any macromolecule (including virtually all proteins or peptides) can be used as an antigen. Antigens and / or epitopes can be expressed endogenously, i.e., through genomic DNA, or they can be recombinantly expressed. Antigens and / or epitopes can be specific to a particular tissue, such as diseased cells, such as cancer cells, or they can be widely expressed. Furthermore, fragments of larger molecules can act as antigens. In one respect, the antigen is a tumor antigen. Epitopes can be present in longer polypeptides (e.g., proteins), or epitopes can exist as fragments of longer polypeptides. In some respects, epitopes are complexed with the major histocompatibility complex (MHC) (also referred to herein as complexed with HLA molecules, such as HLA class 1 molecules).

[0086] The term "autologous" refers to any material derived from the same individual and subsequently reintroduced into that individual. For example, autologous T-cell therapy involves administering T cells isolated from the same individual to a subject. The term "allogeneic" refers to any material derived from one individual and subsequently introduced into another individual of the same species. For example, allogeneic T-cell transplantation involves administering T cells obtained from a donor other than the subject to a subject.

[0087] "Cancer" refers to a broad group of diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and growth lead to the formation of malignant tumors that invade adjacent tissues and can also metastasize to distant parts of the body via the lymphatic system or bloodstream. "Cancer" or "cancer tissue" can include tumors. Examples of cancers that can be treated by the methods of the present invention include, but are not limited to, cancers of the immune system, including lymphoma, leukemia, and other white blood cell malignancies. In some aspects, the methods of the present invention can be used to reduce the size of tumors originating from, for example, melanoma, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, rectal cancer, gastric cancer, uterine cancer, lung cancer, Hodgkin's disease, non-Hodgkin's lymphoma (NHL), esophageal cancer, small bowel cancer, urethral cancer, chronic or acute leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia (ALL) (including non-T-cell ALL), chronic lymphocytic leukemia (CLL), bladder cancer, kidney or ureteral cancer, renal pelvis cancer, glioma, squamous cell carcinoma, and combinations of said cancers. A particular cancer may be responsive to chemotherapy or radiation therapy, or the cancer may be refractory. Refractory cancer is defined as cancer for which surgical intervention is not applicable, and said cancer initially did not respond to chemotherapy or radiation therapy, or said cancer has become unresponsive over time.

[0088] As used herein, the term “progression-free survival” may be abbreviated as PFS, which refers to the time from the date of treatment to the date of disease progression or death from any cause according to the revised IWG criteria for response to malignant lymphoma.

[0089] The term "overall survival" can be abbreviated as OS, and is defined as the time from the date of treatment to the date of death.

[0090] As used herein, the term "infection" refers to any type of invasion of one or more tissues of the body by a foreign substance. The term "infection" includes, but is not limited to, infections caused by viruses (including viroids and prions), bacteria, fungi, parasites, and any combination thereof.

[0091] As used herein, the term “lymphocyte” includes natural killer (NK) cells, T cells, or B cells. NK cells are a type of cytotoxic / cell toxic lymphocyte that represents a major component of the innate immune system. NK cells repel tumor cells and cells infected by viruses. They function through apoptosis, or programmed cell death. They are called “natural killers” because they can kill cells without activation. T cells play a major role in cell-mediated immunity (without antibody involvement). The T cell receptor (TCR) distinguishes T cells from other lymphocyte types. The thymus is a specialized organ of the immune system, primarily responsible for the maturation of T cells. There are six types of T cells: helper T cells (e.g., CD4+ cells), cytotoxic T cells (also known as TC, cytotoxic T lymphocytes, CTL, T killer cells, cytolytic T cells, CD8+ T cells, or killer T cells), memory T cells (i.e., stem memory T cells), and memory T cells. SCM Cells, like immature cells, are CD45RO-, CCR7+, CD45RA+, CD62L+ (L-selectin), CD27+, CD28+, and IL-7Rα+, but they also express large amounts of CD95, IL-2Rβ, CXCR3, and LFA-1, and exhibit many functional properties unique to memory cells; (ii) central memory T cells CM Cells express L-selectin and CCR7; they secrete IL-2 but not IFNγ or IL-4, and (iii) effector memory T cells. EM B cells, however, do not express L-selectin or CCR7 but produce effector cytokines such as IFNγ and IL-4, regulatory T cells (Treg, suppressor T cells, or CD4+CD25+ regulatory T cells), natural killer T cells (NKT), and γδ T cells. On the other hand, B cells play a major role in humoral immunity (with antibody involvement). B cells produce antibodies and antigens and act as antigen-presenting cells (APCs), transforming into memory B cells upon activation by antigen-antigen interactions. In mammals, immature B cells form in the bone marrow, from which their name originates.

[0092] The terms “modified” and “mutated” as used herein to refer to a nucleotide or amino acid sequence mean a change in sequence relative to a wild-type sequence or a specific reference sequence. Unless otherwise stated, the terms “modified” and “mutated” do not require steps in the process of preparing a modified or mutated sequence (e.g., a modified β-chain sequence). Rather, these terms indicate a variation in the modified or mutated sequence relative to a reference sequence (e.g., a wild-type sequence). For example, a DQβ chain containing a substitutional mutation at position 114 corresponding to amino acid residue 114 of SEQ ID NO:1 does not require that the wild-type DQβ chain has been physically altered to obtain the DQβ chain; rather, when properly aligned, the DQβ chain contains amino acid residues at said position (residue 114) that are different from the amino acid residues at the corresponding position in the wild-type or reference DQβ chain.

[0093] As used herein, the term "any amino acid" refers to any known amino acid. An amino acid is an organic compound containing (i) an amine (-NH2) functional group, (ii) a carboxyl (-COOH) functional group, and (iii) a side chain (R group), wherein the side chain is specific to each amino acid. This includes, but is not limited to, any naturally occurring amino acid and any modifications and variants thereof. There are approximately 500 naturally occurring amino acids, of which 20 are encoded by the genetic code. Amino acids with positively charged side chains include arginine (Arg; R), histidine (His; H), and lysine (Lys; K). Amino acids with negatively charged side chains include aspartic acid (Asp; D) and glutamic acid (Glu; E). Amino acids with polar, uncharged side chains include serine (Ser; S), threonine (Thr; T), glutamine (Gln; Q), and asparagine (Asn; N). Amino acids with hydrophobic side chains include alanine (Ala; A), isoleucine (Ile; I), leucine (Leu; L), methionine (Met; M), phenylalanine (Phe; F), valine (Val; V), tryptophan (Trp; W), and tyrosine (Tyr; Y). Tryptophan (Trp; W), tyrosine (Tyr; Y), and methionine (Met; M) can also be classified as polar and / or amphiphilic amino acids because these amino acids are commonly found on the surface of protein or lipid membranes. Other amino acids include cysteine ​​(Cys; C), selenocysteine ​​(Sec; U), glycine (Gly; G), and proline (Pro; P).

[0094] As used herein, “at the position corresponding to…” is a means of identifying a specific amino acid residue (e.g., a specific amino acid position) in a polynucleotide or a specific nucleic acid (e.g., a specific nucleic acid position) in a polypeptide. The position can be determined by correctly aligning the sequence in question with a reference sequence. Those skilled in the art will readily understand how to align sequences to determine relative positions. For example, various alignment tools are available online, including but not limited to “Clustal Omega Multiple Sequence Alignment” available at www.ebi.ac.uk (last accessed May 25, 2019).

[0095] The terms "genetically engineered" or "engineered" refer to methods of modifying the genome of cells, including but not limited to deleting coding or non-coding regions or portions thereof, or inserting coding regions or portions thereof. In some aspects, the modified cells are lymphocytes, such as CD4-expressing T cells, or modified cells that can be obtained from a patient or donor. Cells can be modified to express exogenous constructs, such as the T-cell receptor (TCR) disclosed herein, which are incorporated into the cell genome. In some aspects, cells are modified to express CD4.

[0096] "Immune response" refers to the action of cells of the immune system (e.g., T lymphocytes, B lymphocytes, natural killer (NK) cells, macrophages, eosinophils, mast cells, dendritic cells, and neutrophils) and soluble macromolecules (including alpha-1, cytokines, and complement) produced by any of these cells or the liver, which cause selective targeting, binding, damage, destruction, and / or elimination of invading pathogens, pathogen-infecting cells or tissues, cancer cells or other abnormal cells, or (in the case of autoimmune or pathological inflammation) normal human cells or tissues in vertebrates.

[0097] The term "immunotherapy" refers to the treatment of a subject who has a disease or is at risk of contracting or relapsing into a disease by means of methods including inducing, enhancing, suppressing, or otherwise modifying an immune response. Examples of immunotherapy include, but are not limited to, T-cell therapy. T-cell therapy may include adoptive T-cell therapy, tumor-infiltrating lymphocyte (TIL) immunotherapy, autologous cell therapy, engineered autologous cell therapy (eACT), and allogeneic T-cell transplantation.

[0098] The cells used in the immunotherapy described herein can be derived from any source known in the art. For example, T cells can be differentiated in vitro from hematopoietic stem cell populations, or T cells can be obtained from a subject. T cells can be obtained from, for example, peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymus tissue, tissue from sites of infection, ascites, pleural effusion, spleen tissue, and tumors. Furthermore, T cells can be derived from one or more T cell lines available in the art. T cells can also be generated using any number of techniques known to those skilled in the art (such as FICOLL). TM Isolation and / or apheresis are obtained from units of blood collected from the subject. Further methods for isolating T cells for T-cell therapy are disclosed in U.S. Patent Publication No. 2013 / 0287748, which is incorporated herein by reference in its entirety. Immunotherapy may also include administering modified cells to the subject, wherein said modified cells express the CD4 and TCR disclosed herein. In some aspects, the modified cells are not T cells.

[0099] As used herein, “patient” includes anyone who has cancer (e.g., lymphoma or leukemia). The terms “subject” and “patient” are used interchangeably in this document.

[0100] The terms “peptide,” “polypeptide,” and “protein” are used interchangeably and refer to compounds composed of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and there is no limit to the maximum number of amino acids that can form the sequence of a protein or peptide. A polypeptide includes any peptide or protein containing two or more amino acids linked together by peptide bonds. As used herein, the term refers both to short chains, such as those commonly referred to in the art as peptides, oligopeptides, and oligomers, and to longer chains, which are commonly referred to in the art as proteins, among many types. “Polypeptide” includes, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, etc. Polypeptides include natural peptides, recombinant peptides, synthetic peptides, or combinations thereof.

[0101] As used herein, “stimulus” refers to a primary response induced by the binding of a stimulating molecule to its homologous ligand, wherein this binding mediates a signal transduction event. A “stimulating molecule” is a molecule on T cells, such as the T cell receptor (TCR) / CD4 complex, which specifically binds to a homologous stimulating ligand present on antigen-presenting cells. A “stimulating ligand” is a stimulating molecule on T cells that, when present on antigen-presenting cells (e.g., aAPCs, dendritic cells, B cells, etc.), can specifically bind to the stimulating molecule on T cells, thereby mediating primary T cell responses, including but not limited to activation, initiation of an immune response, and proliferation. Stimulating ligands include, but are not limited to, MHC class II molecules loaded with peptides, anti-CD4 antibodies, anti-CD28 antibodies, anti-CD2 antibodies, and anti-CD3 antibodies.

[0102] "Treatment" in a subject refers to any type of intervention or treatment administered to the subject, or the administration of an active agent, with the aim of reversing, alleviating, improving, suppressing, slowing, or preventing the onset, progression, development, severity, or recurrence of symptoms, complications, or symptom-related biochemical markers. In one aspect, "treatment" includes partial remission. In another aspect, "treatment" includes complete remission.

[0103] The use of alternatives (e.g., "or") should be understood to mean one, both, or any combination of the alternatives. As used herein, the indefinite article "a / an" should be understood to mean "one or more / a" of any of the described or enumerated components.

[0104] The terms “about” or “substantially comprise” refer to a value or composition within an acceptable margin of error for a particular value or composition as determined by a person skilled in the art, in part depending on how the value or composition is measured or determined, i.e., the limitations of the measurement system. For example, “about” or “substantially comprise” may mean within one or more standard deviations according to practice in the art. Alternatively, “about” or “substantially comprise” may refer to a range up to 10% (i.e., ±10%). For example, about 3 mg may include any amount between 2.7 mg and 3.3 mg (10%). Furthermore, particularly with respect to biological systems or processes, these terms may refer to values ​​up to an order of magnitude or up to five times the value. When a particular value or composition is provided in this application and claims, unless otherwise stated, it should be assumed that the meaning of “about” or “substantially comprise” is within an acceptable margin of error for that particular value or composition.

[0105] As stated herein, unless otherwise indicated, any concentration range, percentage range, ratio range, or integer range shall be understood to include any integer value within the range, and, where appropriate, to include fractions thereof (such as one-tenth and one-hundredth of an integer).

[0106] Various aspects of the invention are described in more detail in the following sections.

[0107] II. Compositions disclosed herein

[0108] This disclosure relates to HLA class II molecules having enhanced CD4 binding. Certain aspects of this disclosure relate to HLA class II molecules comprising a β-chain, wherein said β-chain contains one or more mutations. In some aspects, one or more mutations in the β-chain increase the affinity of the β-chain for CD4. In some aspects, the β-chain is an HLA-DQ (“DQ”) β-chain.

[0109] Class II.A. MHC II molecules

[0110] The human leukocyte antigen (HLA) system (the major histocompatibility complex [MHC] in humans) is an important component of the immune system and is controlled by genes located on chromosome 6. It encodes cell surface molecules on T cells that specifically present antigenic peptides to T cell receptors (TCRs). (See also Overview of the Immune System). Antigen-presenting MHC molecules are divided into two main classes: class I MHC molecules and class II MHC molecules.

[0111] Class II MHC molecules exist as transmembrane glycoproteins on the surface of professional antigen-presenting cells (APCs). A complete class II molecule consists of an α-chain and a β-chain. Three loci in the HLA complex encode MHC class II proteins: HLA-DP, HLA-DQ, and HLA-DR. T cells expressing CD4 molecules respond to class II MHC molecules. These lymphocytes typically have effector and helper functions, activating responses to eliminate intracellular pathogens from their own cells or to destroy extracellular parasites and assist other T cells, such as CD8 T cells. Because only professional APCs express class II MHC molecules, only these cells present antigens to CD4 T cells (CD4 binds to the non-polymorphic portions of the α-2 and β-2 domains of the α and β chains of the class II MHC molecules, respectively).

[0112] In some respects, the HLA class II α and β chains are selected from the HLA-DP, HLA-DQ, and HLA-DR alleles. In some respects, the HLA class II β chain is the HLA-DQ allele. In some respects, the HLA class II α chain is the HLA-DQ allele.

[0113] Many HLA-DQ alleles are known in the art, and any known allele can be used in this disclosure. Examples of HLA-DQ α-chain and β-chain alleles are shown in Table 1. An updated list of HLA alleles is available at hla.alleles.org / (last accessed July 10, 2019).

[0114] II.A.1. MHC Class II β chain

[0115] In some aspects, HLA class II molecules comprise a DQβ chain, wherein the DQβ chain comprises an amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1. Any amino acid other than leucine may be present at position 114 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than leucine is an amino acid containing a hydrophobic side chain. In some aspects, the amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1 is an amino acid selected from alanine, valine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, the amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1 is alanine. In some aspects, the amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1 is valine. In some aspects, the amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1 is isoleucine. In some aspects, the amino acid at position 114 of amino acid residue SEQ ID NO:1, other than leucine, is methionine. In some aspects, the amino acid at position 114 of amino acid residue SEQ ID NO:1, other than leucine, is phenylalanine. In some aspects, the amino acid at position 114 of amino acid residue SEQ ID NO:1, other than leucine, is tyrosine. In some aspects, the amino acid at position 114 of amino acid residue SEQ ID NO:1, other than leucine, is tryptophan.

[0116] In some embodiments, the amino acid at position 114 corresponding to amino acid residue 114 of SEQ ID NO:1, excluding leucine, consists of more than one amino acid, for example, two, three, four, five, or more amino acids. In some aspects, at least one of the more than one amino acid contains a hydrophobic side chain. In some aspects, the amino acid at position 114 corresponding to amino acid residue 114 of SEQ ID NO:1, excluding leucine, consists of a series, for example, at least two, at least three, at least four, or at least five amino acids, wherein each of said series of amino acids contains a hydrophobic side chain.

[0117] In some aspects, HLA class II molecules comprise a DQβ chain, wherein the DQβ chain comprises an amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1. Any amino acid other than valine may be present at position 143 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than valine is an amino acid containing a hydrophobic side chain. In some aspects, the amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1 is an amino acid selected from alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, the amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1 is alanine. In some aspects, the amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1 is isoleucine. In some aspects, the amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1 is leucine. In some aspects, the amino acid at position 143 of amino acid residue SEQ ID NO:1, other than valine, is methionine. In some aspects, the amino acid at position 143 of amino acid residue SEQ ID NO:1, other than valine, is phenylalanine. In some aspects, the amino acid at position 143 of amino acid residue SEQ ID NO:1, other than valine, is tyrosine. In some aspects, the amino acid at position 143 of amino acid residue SEQ ID NO:1, other than valine, is tryptophan.

[0118] In some aspects, the amino acid at position 143 of amino acid residue 143 corresponding to SEQ ID NO:1, excluding valine, consists of more than one amino acid, for example, two, three, four, five, or more amino acids. In some aspects, at least one of the more than one amino acid contains a hydrophobic side chain. In some aspects, the amino acid at position 143 of amino acid residue 143 corresponding to SEQ ID NO:1, excluding valine, consists of a series, for example, at least two, at least three, at least four, or at least five amino acids, wherein each of said series of amino acids contains a hydrophobic side chain.

[0119] In some aspects, HLA class II molecules comprise a DQβ chain, wherein the DQβ chain comprises an amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1. Any amino acid other than asparagine may be present at position 110 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than asparagine is an amino acid comprising a polar, uncharged side chain. In some aspects, the amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1 is an amino acid selected from serine, threonine, and glutamine. In some aspects, the amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1 is serine. In some aspects, the amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1 is threonine. In some aspects, the amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1 is glutamine.

[0120] In some aspects, the amino acid at position 110 of amino acid residue 110 corresponding to SEQ ID NO:1, excluding asparagine, consists of more than one amino acid, for example, two, three, four, five, or more amino acids. In some aspects, at least one of the more than one amino acid contains a polar, uncharged side chain. In some aspects, the amino acid at position 110 of amino acid residue 110 corresponding to SEQ ID NO:1, excluding asparagine, consists of a series, for example, at least two, at least three, at least four, or at least five amino acids, wherein each of said series of amino acids contains a polar, uncharged side chain.

[0121] In some aspects, HLA class II molecules comprise a DQβ chain, wherein the DQβ chain comprises an amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1. Any amino acid other than isoleucine may be present at position 116 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than isoleucine is an amino acid containing a hydrophobic side chain. In some aspects, the amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1 is an amino acid selected from alanine, valine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, the amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1 is alanine. In some aspects, the amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1 is valine. In some aspects, the amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1 is leucine. In some aspects, the amino acid at position 116 of SEQ ID NO:1, excluding isoleucine, is methionine. In some aspects, the amino acid at position 116 of SEQ ID NO:1, excluding isoleucine, is phenylalanine. In some aspects, the amino acid at position 116 of SEQ ID NO:1, excluding isoleucine, is tyrosine. In some aspects, the amino acid at position 116 of SEQ ID NO:1, excluding isoleucine, is tryptophan.

[0122] In some aspects, the amino acid at position 116 corresponding to amino acid residue 116 of SEQ ID NO:1, excluding isoleucine, consists of more than one amino acid, for example, two, three, four, five, or more amino acids. In some aspects, at least one of the more than one amino acid contains a hydrophobic side chain. In some aspects, the amino acid at position 116 corresponding to amino acid residue 116 of SEQ ID NO:1, excluding isoleucine, consists of a series, for example, at least two, at least three, at least four, or at least five amino acids, wherein each of said series of amino acids contains a hydrophobic side chain.

[0123] In some aspects, HLA class II molecules comprise a DQβ chain, wherein the DQβ chain comprises an amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1. Any amino acid other than serine may be present at position 118 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than serine is an amino acid containing a charged side chain. In some aspects, the amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1 is an amino acid selected from arginine, histidine, and lysine. In some aspects, the amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1 is arginine. In some aspects, the amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1 is histidine. In some aspects, the amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1 is lysine.

[0124] In some aspects, the amino acid at position 118 corresponding to amino acid residue 118 of SEQ ID NO:1, excluding serine, consists of more than one amino acid, for example, two, three, four, five, or more amino acids. In some aspects, at least one of the more than one amino acid contains a charged side chain. In some aspects, the amino acid at position 118 corresponding to amino acid residue 118 of SEQ ID NO:1, excluding serine, consists of a series, for example, at least two, at least three, at least four, or at least five amino acids, wherein each of said series of amino acids contains a charged side chain.

[0125] In some aspects, HLA class II molecules comprise a DQβ chain, wherein the DQβ chain comprises an amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1. Any amino acid other than proline may be present at position 146 of amino acid residue SEQ ID NO:1. In some aspects, the amino acid other than proline is an amino acid comprising a polar, uncharged side chain. In some aspects, the amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1 is an amino acid selected from serine, threonine, asparagine, and glutamine. In some aspects, the amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1 is serine. In some aspects, the amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1 is threonine. In some aspects, the amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1 is asparagine. In some respects, the amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1 is glutamine.

[0126] In some aspects, the amino acid at position 146 corresponding to amino acid residue 1 of SEQ ID NO:1, excluding proline, consists of more than one amino acid, for example, two, three, four, five, or more amino acids. In some aspects, at least one of the more than one amino acid contains a polar, uncharged side chain. In some aspects, the amino acid at position 146 corresponding to amino acid residue 1 of SEQ ID NO:1, excluding proline, consists of a series, for example, at least two, at least three, at least four, or at least five amino acids, wherein each of said series of amino acids contains a polar, uncharged side chain.

[0127] In some aspects of this disclosure, MHC class II molecules comprise a DQβ chain containing more than one substitution mutation relative to the wild-type DQβ chain. In some aspects, the DQβ chain contains at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, or at least ten mutations relative to the wild-type DQβ chain.

[0128] In some aspects, the DQβ chain comprises an amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1 and an amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1. In some aspects, the DQβ chain comprises an amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1; an amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1; and at least three of the following: (i) an amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1, (ii) an amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1, (iii) an amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1, and (iv) an amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1.

[0129] In some respects, the DQβ chain comprises (i) an amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1; (ii) an amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1; (iii) an amino acid other than asparagine at position 110 of amino acid residue SEQ ID NO:1; (iv) an amino acid other than isoleucine at position 116 of amino acid residue SEQ ID NO:1; (v) an amino acid other than serine at position 118 of amino acid residue SEQ ID NO:1; and (vi) an amino acid other than proline at position 146 of amino acid residue SEQ ID NO:1.

[0130] In some respects, (i) the amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1, (ii) the amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1, or each of the amino acid other than leucine at position 114 of amino acid residue SEQ ID NO:1 and the amino acid other than valine at position 143 of amino acid residue SEQ ID NO:1 is an amino acid containing a hydrophobic side chain.

[0131] In some respects, (i) the amino acid at position 114 of SEQ ID NO:1, excluding leucine, is selected from alanine, valine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan; (ii) the amino acid at position 143 of SEQ ID NO:1, excluding valine, is selected from alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan; (iii) the amino acid at position 110 of SEQ ID NO:1, excluding asparagine, is selected from serine, threonine, and glutamine; (iv) the amino acid at position 116 of SEQ ID NO:1, excluding isoleucine, is selected from alanine, valine, leucine, methionine, phenylalanine, tyrosine, and tryptophan; (v) the amino acid at position 118 of SEQ ID NO:1, excluding serine, is selected from arginine, histidine, and lysine; and (vi) the amino acid at position 143 of SEQ ID NO:1, excluding valine, is selected from alanine, valine, leucine, methionine, phenylalanine, tyrosine, and tryptophan; The amino acid at position 146 of NO:1, excluding proline, is selected from serine, threonine, asparagine, and glutamine.

[0132] In some aspects, (i) the amino acid at position 114 of amino acid residue SEQ ID NO:1, excluding leucine, is tryptophan; and (ii) the amino acid at position 143 of amino acid residue SEQ ID NO:1, excluding valine, is selected from alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, (i) the amino acid at position 114 of amino acid residue SEQ ID NO:1, excluding leucine, is selected from alanine, valine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan; and (ii) the amino acid at position 143 of amino acid residue SEQ ID NO:1, excluding valine, is methionine. In some aspects, (i) the amino acid at position 114 of amino acid residue SEQ ID NO:1, excluding leucine, is tryptophan; and (ii) the amino acid at position 143 of amino acid residue SEQ ID NO:1, excluding valine, is methionine. In some respects, (i) the amino acid at position 114 of SEQ ID NO:1, excluding leucine, is tryptophan; (ii) the amino acid at position 143 of SEQ ID NO:1, excluding valine, is methionine; (iii) the amino acid at position 110 of SEQ ID NO:1, excluding asparagine, is glutamine; (iv) the amino acid at position 116 of SEQ ID NO:1, excluding isoleucine, is valine; (v) the amino acid at position 118 of SEQ ID NO:1, excluding serine, is histidine; and (vi) the amino acid at position 146 of SEQ ID NO:1, excluding proline, is glutamine.

[0133] In some respects, the DQβ chain described herein exhibits increased affinity for CD4 protein compared to a reference HLA class II molecule. In some respects, the reference HLA class II molecule is an HLA class II molecule possessing a wild-type DQβ chain. In some respects, the reference HLA class II molecule is an HLA class II molecule possessing a DQβ chain comprising (i) a leucine residue at position 114 corresponding to amino acid residue SEQ ID NO:1 and / or (ii) a valine residue at position 143 corresponding to amino acid residue SEQ ID NO:1. In some respects, the reference HLA class II molecule is an HLA class II molecule having a DQβ chain comprising (i) leucine at position 114 of amino acid residue SEQ ID NO:1, (ii) valine at position 143 of amino acid residue SEQ ID NO:1, (iii) asparagine at position 110 of amino acid residue SEQ ID NO:1, (iv) isoleucine at position 116 of amino acid residue SEQ ID NO:1, (iii) serine at position 118 of amino acid residue SEQ ID NO:1, and / or (iv) proline at position 146 of amino acid residue SEQ ID NO:1.

[0134] In some respects, the increased affinity for CD4 is at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 15 times, at least about 20 times, at least about 25 times, at least about 30 times, at least about 35 times, at least about 40 times, at least about 45 times, at least about 50 times, at least about 75 times, at least about 100 times, at least about 200 times, at least about 300 times, at least about 400 times, at least about 500 times, at least about 1000 times, at least about 1500 times, at least about 2000 times, at least about 2500 times, at least about 3000 times, at least about 3500 times, at least about 4000 times, at least about 4500 times, or at least about 4000 times.

[0135] In some respects, the increased affinity for CD4 is at least approximately 1.5 to at least approximately 5000 times, 1.5 to at least approximately 4000 times, 1.5 to at least approximately 3000 times, 1.5 to at least approximately 2000 times, 1.5 to at least approximately 1000 times, 10 to at least approximately 5000 times, 10 to at least approximately 4000 times, 10 to at least approximately 3000 times, 10 to at least approximately 2000 times, 10 to at least approximately 1000 times, 10 to at least approximately 900 times, 10 to at least approximately 800 times, 10 to at least approximately 700 times, 10 to at least approximately 600 times, 10 to at least approximately 500 times, and 10 to at least approximately 500 times, respectively. At least 400 times, 10 times to at least 300 times, 10 times to at least 200 times, 10 times to at least 100 times, 100 times to at least 5000 times, 100 times to at least 4000 times, 100 times to at least 3000 times, 100 times to at least 2000 times, 100 times to at least 1000 times, 100 times to at least 900 times, 100 times to at least 800 times, 100 times to at least 700 times, 100 times to at least 600 times, 100 times to at least 500 times, 100 times to at least 400 times, 100 times to at least 300 times, or 100 times to at least 200 times.

[0136] In some respects, the DQβ chain contains alleles selected from the following: HLA-DQB1*02, HLA-DQB1*03, HLA-DQB1*04, HLA-DQB1*05, and HLA-DQB1*06. In some respects, the DQβ chain contains the HLA-DQB1*05 allele. In certain respects, the DQβ chain contains the HLA-DQB1*05:01 allele.

[0137] In some respects, the DQβ chain contains alleles selected from the following: DQB1*02:01:01, DQB1*02:01:02, DQB1*02:01:03, DQB1*02:01:04, DQB1*02:01:05, DQB1*02:01:06, DQB1*02:01:07, DQB1*02:01:08, DQB1*02:01:09, DQB1*02:01:10, DQB1*02:01:11, DQB1*02:01:12, DQB1*02:01:13, DQB1*02:01:14, DQB1*02:01:15, DQB1*02:01:16, DQB 1*02:01:17, DQB1*02:01:18, DQB1*02:01:19, DQB1*02:01:20, DQB1*02:0 1:21, DQB1*02:01:22, DQB1*02:01:23, DQB1*02:01:24, DQB1*02:01:25, D QB1*02:01:26, DQB1*02:01:27, DQB1*02:01:28, DQB1*02:01:29, DQB1*02 :01:30, DQB1*02:01:31, DQB1*02:02:01:01, DQB1*02:02:01:02, DQB1*02: 02:01:03, DQB1*02:02:01:04, DQB1*02:02:02, DQB1*02:02:03, DQB1*02: 02:04, DQB1*02:02:05, DQB1*02:02:06, DQB1*02:02:07, DQB1*02:02:08, DQB1*02:02:09, DQB1*02:03:01, DQB1*02:03:02, DQB1*02:04, DQB1*02:0 5. DQB1*02:06, DQB1*02:07:01, DQB1*02:07:02, DQB1*02:08, DQB1*02:09, DQB1*02:10, DQB1*02:100, DQB1*02:101, DQB1*02:102, DQB1*02:103, DQB 1*02:104, DQB1*02:105, DQB1*02:106, DQB1*02:107, DQB1*02:108, DQB1*0 2:109, DQB1*02:11, DQB1*02:110, DQB1*02:111, DQB1*02:112, DQB1*02:1 13. DQB1*02:114, DQB1*02:115, DQB1*02:116, DQB1*02:117, DQB1*02:118,DQB1*02:119、DQB1*02:12、DQB1*02:120、DQB1*02:121、DQB1*02:122、DQB1*02:123、DQB1*02:124、DQB1*02:125、DQB1*02:126、DQB1*02:127、DQB1*02:128、DQB1*02:129N、DQB1*02:13、DQB1*02:130、DQB1*02:131、DQB1*02:132N、DQB1*02:133、DQB1*02:134N、DQB1*02:135、DQB1*02:136、DQB1*02: 137、DQB1*02:138、DQB1*02:139、DQB1*02:140、DQB1*02:141、DQB1*02:142、DQB1*02:14:01、DQB1*02:14:02、DQB1*02:15、DQB1*02:16、DQB1*02:17、DQB1*02:18N、DQB1*02:19、DQB1*02:20N、DQB1*02:21、DQB1*02:22、DQB1*02:23、DQB1*02:24、DQB1*02:25、DQB1*02:26、DQB1*02:27、DQB1*02:28、D QB1*02:29、DQB1*02:30、DQB1*02:31、DQB1*02:32、DQB1*02:33、DQB1*02:34、DQB1*02:35、DQB1*02:36、DQB1*02:37、DQB1*02:38、DQB1*02:39、DQB1*02:40、DQB1*02:41、DQB1*02:42、DQB1*02:43、DQB1*02:44、DQB1*02:45、DQB1*02:46、DQB1*02:47、DQB1*02:48、DQB1*02:49、DQB1*02:50、DQB1*02: 51、DQB1*02:52、DQB1*02:53Q、DQB1*02:54、DQB1*02:55、DQB1*02:56、DQB1*02:57、DQB1*02:58N、DQB1*02:59、DQB1*02:60、DQB1*02:61、DQB1*02:62、DQB1*02:63、DQB1*02:64、DQB1*02:65、DQB1*02:66、DQB1*02:67NX、DQB1*02:68、DQB1*02:69、DQB1*02:70、DQB1*02:71、DQB1*02:72、DQB1*02:73、DQB1*02:74、DQB1*02:75、DQB1*02:76、DQB1*02:77、DQB1*02:78、DQB1*02:79、DQB1*02:80、DQB1*02:81、DQB1*02:82、DQB1*02:83、DQB1*02:84、DQB1*02:85、DQB1*02:86、DQB1*02:87、DQB1*02:88、DQB1*02:89:01、DQB1*02:89:02、DQB1*02:90、DQB1*02:91、DQB1*02:92、DQB1*02:93、DQB1*02:94 ,DQB1*02:95,DQB1*02:96N,DQB1*02:97,DQB1*02:98,DQB1*02:99,DQB1*03:01:01:01,DQB1*03:01:01:02,DQB1*03:01:01:03,DQB1*03:01:01:04,DQB1*03:01:01:05,DQB1*03:01:01:06,DQB1*03:01:01:07,DQB1*03:01:01:08,DQB1*03:01:01:09,DQB1*03:01:01:10,DQB1*03:01:01:11,DQB1 *03:01:01:12、DQB1*03:01:01:14、DQB1*03:01:01:15、DQB1*03:01:01:16、DQB1*03:01:01:17、DQB1*03:01:01:18、DQB1*03:01:01:19、DQB1*03:01:01:20、DQB1*03:01:02、DQB1*03:01:03、DQB1*03:01:04、DQB1*03:01:05、DQB1*03:01:06、DQB1*03:01:07、DQB1*03:01:08、DQB1*03:01:09、DQB1 *03:01:10、DQB1*03:01:11、DQB1*03:01:12、DQB1*03:01:13、DQB1*03:01:14、DQB1*03:01:15、DQB1*03:01:16、DQB1*03:01:17、DQB1*03:01:18、DQB1*03:01:19、DQB1*03:01:20、DQB1*03:01:21、DQB1*03:01:22、DQB1*03:01:23、DQB1*03:01:24、DQB1*03:01:25、DQB1*03:01:26、DQB1*03:01:27、DQB1*03:01:28、DQB1*03:01:29、DQB1*03:01:30、DQB1*03:01:31、DQB1*03:01:32、DQB1*03:01:33、DQB1*03:01:34、DQB1*03:01:35、DQB1*03:01:36、DQB1*03:01:37、DQB1*03:01:38、DQB1*03:01:39、DQB1*03:01:40、DQB1*03:01:41、DQB1*03:01:42、DQB1*03:01:43、DQB1*03:01:44、DQB1*03:0 1:45、DQB1*03:01:46、DQB1*03:02:01:01、DQB1*03:02:01:02、DQB1*03:02:01:03、DQB1*03:02:01:04、DQB1*03:02:01:05、DQB1*03:02:01:06、DQB1*03:02:01:07、DQB1*03:02:01:08、DQB1*03:02:02、DQB1*03:02:03、DQB1*03:02:04、DQB1*03:02:05、DQB1*03:02:06、DQB1*03:02:07、DQB1*03:0 2:08、DQB1*03:02:09、DQB1*03:02:10、DQB1*03:02:11、DQB1*03:02:12、DQB1*03:02:13、DQB1*03:02:14、DQB1*03:02:15、DQB1*03:02:16、DQB1*03:02:17、DQB1*03:02:18、DQB1*03:02:19、DQB1*03:02:20、DQB1*03:02:21、DQB1*03:02:22、DQB1*03:02:23、DQB1*03:02:24、DQB1*03:02:25、DQB1* 03:02:26、DQB1*03:02:27、DQB1*03:02:28、DQB1*03:02:29、DQB1*03:02:30、DQB1*03:03:02:01、DQB1*03:03:02:02、DQB1*03:03:02:03、DQB1*03:03:02:04、DQB1*03:03:02:05、DQB1*03:03:03、DQB1*03:03:04、DQB1*03:03:05、DQB1*03:03:06、DQB1*03:03:07、DQB1*03:03:08、DQB1*03:03:09、DQB1*03:03:10、DQB1*03:03:11、DQB1*03:03:12、DQB1*03:03:13、DQB1*03:03:14、DQB1*03:03:15、DQB1*03:03:16、DQB1*03:03:17、DQB1*03:03:18、DQB1*03:03:19、DQB1*03:03:20、DQB1*03:03:21、DQB1*03:04:01、DQB1*03:04:02、DQB1*03:04:03、DQB1*03:04:04、DQB1*03:05:01、DQB1*03:05 :02、DQB1*03:05:03、DQB1*03:05:04、DQB1*03:06、DQB1*03:07、DQB1*03:08、DQB1*03:09、DQB1*03:100、DQB1*03:101、DQB1*03:102、DQB1*03:103、DQB1*03:104、DQB1*03:105、DQB1*03:106、DQB1*03:107、DQB1*03:108、DQB1*03:109、DQB1*03:10:01、DQB1*03:10:02:01、DQB1*03:10:02:02、DQB1* 03:11、DQB1*03:110、DQB1*03:111、DQB1*03:112、DQB1*03:113、DQB1*03:114、DQB1*03:115、DQB1*03:116、DQB1*03:117、DQB1*03:118N、DQB1*03:119、DQB1*03:12、DQB1*03:120、DQB1*03:121、DQB1*03:122、DQB1*03:123、DQB1*03:124、DQB1*03:125、DQB1*03:126、DQB1*03:127、DQB1*03:128、DQ B1*03:129、DQB1*03:13、DQB1*03:130、DQB1*03:131、DQB1*03:132、DQB1*03:133、DQB1*03:134、DQB1*03:135、DQB1*03:136、DQB1*03:137、DQB1*03:138、DQB1*03:139、DQB1*03:140、DQB1*03:141、DQB1*03:142、DQB1*03:143、DQB1*03:144、DQB1*03:145、DQB1*03:146、DQB1*03:147、DQB1*03:148、DQB1*03:149、DQB1*03:14:01、DQB1*03:14:02、DQB1*03:15、DQB1*03:150、DQB1*03:151、DQB1*03:152、DQB1*03:153、DQB1*03:154、DQB1*03:155、DQB1*03:156、DQB1*03:157、DQB1*03:158、DQB1*03:159、DQB1*03:16、DQB1*03:160、DQB1*03:161、DQB1*03:162、DQB1*03:163、DQB1*03:164、DQB1*0 3:165、DQB1*03:166、DQB1*03:167、DQB1*03:168、DQB1*03:169、DQB1*03:170、DQB1*03:171、DQB1*03:172、DQB1*03:173、DQB1*03:174、DQB1*03:175、DQB1*03:176、DQB1*03:177、DQB1*03:178、DQB1*03:179、DQB1*03:17:01、DQB1*03:17:02、DQB1*03:18、DQB1*03:180、DQB1*03:181、DQB1*03:182 ,DQB1*03:183,DQB1*03:184,DQB1*03:185,DQB1*03:186,DQB1*03:187,DQB1*03:188,DQB1*03:189,DQB1*03:190,DQB1*03:191,DQB1*03:192,DQB1*03:193,DQB1*03:194,DQB1*03:195,DQB1*03:196,DQB1*03:197Q,DQB1*03:198:01,DQB1*03:198:02,DQB1*03:199,DQB1*03:19:01,DQB1*03:19 :02、DQB1*03:19:03、DQB1*03:19:04、DQB1*03:20、DQB1*03:200、DQB1*03:201、DQB1*03:202、DQB1*03:203、DQB1*03:204、DQB1*03:205、DQB1*03:206、DQB1*03:207、DQB1*03:208、DQB1*03:209、DQB1*03:21、DQB1*03:210、DQB1*03:211、DQB1*03:212、DQB1*03:213NX、DQB1*03:214、DQB1*03:215、DQB1*03:216、DQB1*03:217、DQB1*03:218、DQB1*03:219、DQB1*03:220、DQB1*03:221、DQB1*03:222、DQB1*03:223、DQB1*03:224、DQB1*03:225、DQB1*03:226、DQB1*03:227、DQB1*03:228、DQB1*03:229、DQB1*03:22:01、DQB1*03:22:02、DQB1*03:230、DQB1*03:231、DQB1*03:232、DQB1*03:233、DQB1* 03:234、DQB1*03:235、DQB1*03:236、DQB1*03:237N、DQB1*03:238、DQB1*03:239、DQB1*03:23:01、DQB1*03:23:02、DQB1*03:23:03、DQB1*03:24、DQB1*03:240、DQB1*03:241、DQB1*03:242、DQB1*03:243、DQB1*03:244、DQB1*03:245、DQB1*03:246、DQB1*03:247、DQB1*03:248、DQB1*03:249、DQB1*03: 250、DQB1*03:251、DQB1*03:252、DQB1*03:253、DQB1*03:254、DQB1*03:255、DQB1*03:256、DQB1*03:257、DQB1*03:258、DQB1*03:259、DQB1*03:25:01、DQB1*03:25:02、DQB1*03:26、DQB1*03:260、DQB1*03:261、DQB1*03:262、DQB1*03:263、DQB1*03:264、DQB1*03:265、DQB1*03:266、DQB1*03:267、DQ B1*03:268、DQB1*03:269N、DQB1*03:27、DQB1*03:270、DQB1*03:271、DQB1*03:272、DQB1*03:273、DQB1*03:274、DQB1*03:275、DQB1*03:277、DQB1*03:278、DQB1*03:279、DQB1*03:28、DQB1*03:280、DQB1*03:281、DQB1*03:282N、DQB1*03:283、DQB1*03:284、DQB1*03:285、DQB1*03:286、DQB1*03:287、DQB1*03:288、DQB1*03:289、DQB1*03:29、DQB1*03:290、DQB1*03:291、DQB1*03:292、DQB1*03:293、DQB1*03:294、DQB1*03:295、DQB1*03:296、DQB1*03:297、DQB1*03:298、DQB1*03:299、DQB1*03:30、DQB1*03:300、DQB1*03:301、DQB1*03:302、DQB1*03:303N、DQB1*03:304、DQB1*03:305、DQB1*03:30 6、DQB1*03:307、DQB1*03:308、DQB1*03:309、DQB1*03:31、DQB1*03:310N、DQB1*03:311、DQB1*03:312、DQB1*03:313、DQB1*03:314、DQB1*03:315、DQB1*03:316、DQB1*03:317:01、DQB1*03:317:02、DQB1*03:318、DQB1*03:319、DQB1*03:32、DQB1*03:320、DQB1*03:321、DQB1*03:322、DQB1*03:323、DQB 1*03:324、DQB1*03:326、DQB1*03:327、DQB1*03:328、DQB1*03:329、DQB1*03:33、DQB1*03:330、DQB1*03:331、DQB1*03:332、DQB1*03:333、DQB1*03:334N4bp、DQB1*03:335、DQB1*03:336、DQB1*03:337、DQB1*03:338N、DQB1*03:339N、DQB1*03:34、DQB1*03:340N、DQB1*03:341、DQB1*03:342、DQB1*03: 343、DQB1*03:344、DQB1*03:345、DQB1*03:346、DQB1*03:347、DQB1*03:348、DQB1*03:349、DQB1*03:35、DQB1*03:350、DQB1*03:351、DQB1*03:352、DQB1*03:353、DQB1*03:354N、DQB1*03:355、DQB1*03:356NX、DQB1*03:357N、DQB1*03:358N、DQB1*03:36、DQB1*03:37、DQB1*03:38:01、DQB1*03:38:02、DQB1*03:39、DQB1*03:40、DQB1*03:41、DQB1*03:42、DQB1*03:43、DQB1*03:44、DQB1*03:45、DQB1*03:46、DQB1*03:47、DQB1*03:48、DQB1*03:49、DQB1*03:50、DQB1*03:51、DQB1*03:52、DQB1*03:53、DQB1*03:54、DQB1*03:55、DQB1*03:56、DQB1*03:57、DQB1*03:58、DQB1*03:59、DQB1*03:60、DQB1*03: 61、DQB1*03:62、DQB1*03:63、DQB1*03:64、DQB1*03:65、DQB1*03:66N、DQB1*03:67、DQB1*03:68、DQB1*03:69、DQB1*03:70、DQB1*03:71、DQB1*03:72、DQB1*03:73、DQB1*03:74、DQB1*03:75、DQB1*03:76、DQB1*03:77、DQB1*03:78、DQB1*03:79、DQB1*03:80、DQB1*03:81、DQB1*03:82、DQB1*03:83、DQB1 *03:84N、DQB1*03:85、DQB1*03:86、DQB1*03:87、DQB1*03:88、DQB1*03:89、DQB1*03:90N、DQB1*03:91Q、DQB1*03:92、DQB1*03:93、DQB1*03:94、DQB1*03:95N、DQB1*03:96、DQB1*03:97、DQB1*03:98、DQB1*03:99Q、DQB1*04:01:01、DQB1*04:01:01:02、DQB1*04:01:02、DQB1*04:01:03、DQB1*04:01:04: 04、DQB1*04:01:05、DQB1*04:02:01:01、DQB1*04:02:01:04、DQB1*04:02:01:05、DQB1*04:02:01:06、DQB1*04:02:01:07、DQB1*04:02:01:08、DQB1*04:02:01:09、DQB1*04:02:01:10、DQB1*04:02:02、DQB1*04:02:03、DQB1*04:02:04、DQB1*04:02:05、DQB1*04:02:06、DQB1*04:02:07、DQB1*04:02:08、DQB1*04:02:09、DQB1*04:02:10、DQB1*04:02:11、DQB1*04:02:12、DQB1*04:02:13、DQB1*04:02:14、DQB1*04:02:15、DQB1*04:02:16、DQB1*04:02:17、DQB1*04:02:18、DQB1*04:03:01、DQB1*04:03:02、DQB1*04:03、DQB1*04:04、DQB1*04:05、DQB1*04:06、DQB1*04:07、DQB1*04:08、DQB1*04:09 ,DQB1*04:10,DQB1*04:11,DQB1*04:12,DQB1*04:13,DQB1*04:14,DQB1*04:15,DQB1*04:16,DQB1*04:17,DQB1*04:18,DQB1*04:19,DQB1*04:20,DQB1*04:21,DQB1*04:22,DQB1*04:23,DQB1*04:24,DQB1*04:25N,DQB1*04:26,DQB1*04:27,DQB1*04:28,DQB1*04:29,DQB1*04:30,DQB1*04:31,DQB1 *04:32、DQB1*04:33、DQB1*04:34、DQB1*04:35、DQB1*04:36N、DQB1*04:37、DQB1*04:38、DQB1*04:39、DQB1*04:40、DQB1*04:41N、DQB1*04:42、DQB1*04:43、DQB1*04:44、DQB1*04:45、DQB1*04:46N、DQB1*04:47、DQB1*04:48、DQB1*04:49、DQB1*04:50、DQB1*04:51、DQB1*04:52、DQB1*04:53、DQB1*04 :54、DQB1*04:55、DQB1*04:56、DQB1*04:57、DQB1*04:58、DQB1*04:59N、DQB1*04:60、DQB1*04:61、DQB1*04:62、DQB1*05:01:01:01、DQB1*05:01:01:02、DQB1*05:01:01:03、DQB1*05:01:01:04、DQB1*05:01:01:05、DQB1*05:01:02、DQB1*05:01:03、DQB1*05:01:04、DQB1*05:01:05、DQB1*05:01:06、DQB1*05:01:07、DQB1*05:01:08、DQB1*05:01:09、DQB1*05:01:10、DQB1*05:01:11、DQB1*05:01:12、DQB1*05:01:13、DQB1*05:01:14、DQB1*05:01:15、DQB1*05:01:16、DQB1*05:01:17、DQB1*05:01:18、DQB1*05:01:19、DQB1*05:01:20、DQB1*05:01:21、DQB1*05:01:22、DQB1*05:01:23、DQB1*05:01: 24:01、DQB1*05:01:24:02、DQB1*05:01:25、DQB1*05:01:26、DQB1*05:01:27、DQB1*05:01:28、DQB1*05:01:29、DQB1*05:01:30、DQB1*05:01:31、DQB1*05:01:32、DQB1*05:01:33、DQB1*05:01:34、DQB1*05:02:01:01、DQB1*05:02:01:02、DQB1*05:02:01:03、DQB1*05:02:01:04、DQB1*05:02:01:05、DQ B1*05:02:01:06、DQB1*05:02:02、DQB1*05:02:03、DQB1*05:02:04、DQB1*05:02:05、DQB1*05:02:06、DQB1*05:02:07、DQB1*05:02:08、DQB1*05:02:09、DQB1*05:02:10、DQB1*05:02:11、DQB1*05:02:12、DQB1*05:02:13、DQB1*05:02:14、DQB1*05:02:15、DQB1*05:02:16、DQB1*05:02:17、DQB1*05:02: 18、DQB1*05:02:19、DQB1*05:03:01:01、DQB1*05:03:01:02、DQB1*05:03:01:03、DQB1*05:03:02、DQB1*05:03:03、DQB1*05:03:04、DQB1*05:03:05、DQB1*05:03:06、DQB1*05:03:07、DQB1*05:03:08、DQB1*05:03:09、DQB1*05:03:10、DQB1*05:03:11、DQB1*05:03:12、DQB1*05:03:13、DQB1*05:03:14、DQB1*05:03:15、DQB1*05:03:16、DQB1*05:03:17、DQB1*05:03:18、DQB1*05:03:19、DQB1*05:03:20、DQB1*05:04、DQB1*05:05:01、DQB1*05:05:02、DQB1*05:06:01、DQB1*05:06:02、DQB1*05:07、DQB1*05:08、DQB1*05:09、DQB1*05:10、DQB1*05:100、DQB1*05:101、DQB1*05:102、DQB1*05:103、DQB1*05 :104、DQB1*05:105、DQB1*05:106、DQB1*05:107、DQB1*05:108、DQB1*05:109、DQB1*05:110N、DQB1*05:111、DQB1*05:112、DQB1*05:113、DQB1*05:114 、DQB1*05:115、DQB1*05:116、DQB1*05:117、DQB1*05:118、DQB1*05:119、DQB1*05:11:01、DQB1*05:11:02、DQB1*05:12、DQB1*05:120、DQB1*05:121、D QB1*05:122、DQB1*05:123、DQB1*05:124、DQB1*05:125、DQB1*05:126、DQB1*05:127、DQB1*05:128N、DQB1*05:129、DQB1*05:13、DQB1*05:130、DQB1*05:131、DQB1*05:132Q、DQB1*05:133、DQB1*05:134、DQB1*05:135、DQB1*05:136、DQB1*05:137、DQB1*05:138、DQB1*05:139、DQB1*05:14、DQB1*05:140 ,DQB1*05:141,DQB1*05:142,DQB1*05:143,DQB1*05:144,DQB1*05:145,DQB1*05:146,DQB1*05:147,DQB1*05:148,DQB1*05:149,DQB1*05:15,DQB1*05:150,DQB1*05:151,DQB1*05:152,DQB1*05:153,DQB1*05:154,DQB1*05:155,DQB1*05:156,DQB1*05:157,DQB1*05:158,DQB1*05:159,DQB1*05:16,DQB1*05:160、DQB1*05:161、DQB1*05:162、DQB1*05:163、DQB1*05:164、DQB1*05:165、DQB1*05:166、DQB1*05:167、DQB1*05:168、DQB1*05:169、DQB1*05:17、DQB1*05:170、DQB1*05:171、DQB1*05:172、DQB1*05:173、DQB1*05:174、DQB1*05:175、DQB1*05:176、DQB1*05:177、DQB1*05:178、DQB1*05: 179、DQB1*05:18、DQB1*05:180、DQB1*05:181、DQB1*05:182、DQB1*05:183、DQB1*05:184、DQB1*05:185N、DQB1*05:186、DQB1*05:187、DQB1*05:188、DQB1*05:189、DQB1*05:19、DQB1*05:190、DQB1*05:191、DQB1*05:192、DQB1*05:193、DQB1*05:194、DQB1*05:195、DQB1*05:196、DQB1*05:197、DQB1* 05:198、DQB1*05:199、DQB1*05:20、DQB1*05:200、DQB1*05:201、DQB1*05:202、DQB1*05:203、DQB1*05:204、DQB1*05:205、DQB1*05:206N、DQB1*05:207、DQB1*05:208N5bp、DQB1*05:209、DQB1*05:21、DQB1*05:210、DQB1*05:211、DQB1*05:212、DQB1*05:213、DQB1*05:214、DQB1*05:215N、DQB1*05: 216、DQB1*05:217、DQB1*05:22、DQB1*05:23、DQB1*05:24、DQB1*05:25、DQB1*05:26、DQB1*05:27、DQB1*05:28、DQB1*05:29、DQB1*05:30、DQB1*05:31、DQB1*05:32、DQB1*05:33、DQB1*05:34、DQB1*05:35、DQB1*05:36、DQB1*05:37、DQB1*05:38、DQB1*05:39、DQB1*05:40、DQB1*05:41N、DQB1*05:42、DQB1*05:43:01、DQB1*05:43:02、DQB1*05:44、DQB1*05:45、DQB1*05:46、DQB1*05:47、DQB1*05:48、DQB1*05:49、DQB1*05:50、DQB1*05:51、DQB1*05:52、DQB1*05:53、DQB1*05:54、DQB1*05:55、DQB1*05:56、DQB1*05:57、DQB1*05:58、DQB1*05:59、DQB1*05:60、DQB1*05:61、DQB1*05:62、DQB1*05:63、 DQB1*05:64、DQB1*05:65、DQB1*05:66:01、DQB1*05:66:02、DQB1*05:67、DQB1*05:68、DQB1*05:69、DQB1*05:70、DQB1*05:71、DQB1*05:72、DQB1*05:73、DQB1*05:74、DQB1*05:75、DQB1*05:76、DQB1*05:77、DQB1*05:78、DQB1*05:79、DQB1*05:80、DQB1*05:81、DQB1*05:82、DQB1*05:83、DQB1*05:84、D QB1*05:85、DQB1*05:86、DQB1*05:87Q、DQB1*05:88、DQB1*05:89:01、DQB1*05:89:02、DQB1*05:90N、DQB1*05:91、DQB1*05:92、DQB1*05:93、DQB1*05:94、DQB1*05:95、DQB1*05:96、DQB1*05:97、DQB1*05:98、DQB1*05:99、DQB1*06:01:01、DQB1*06:01:02、DQB1*06:01:02、DQB1*06:01:03、DQB 1*06:01:04、DQB1*06:01:05、DQB1*06:01:06、DQB1*06:01:07、DQB1*06:01:08、DQB1*06:01:09、DQB1*06:01:10、DQB1*06:01:11、DQB1*06:01:12、DQB1*06:01:13、DQB1*06:01:14、DQB1*06:01:15、DQB1*06:01:16、DQB1*06:01:17、DQB1*06:01:18、DQB1*06:01:19、DQB1*06:01:20、DQB1*06:01:21、DQB1*06:02:01:01、DQB1*06:02:01:02、DQB1*06:02:01:03、DQB1*06:02:01:04、DQB1*06:02:02、DQB1*06:02:03、DQB1*06:02:04、DQB1*06:02:05、DQB1*06:02:06、DQB1*06:02:07、DQB1*06:02:08、DQB1*06:02:09、DQB1*06:02:10、DQB1*06:02:11、DQB1*06:02:12、DQB1*06:02:13、DQB1*06:02:14、 DQB1*06:02:15、DQB1*06:02:16、DQB1*06:02:17、DQB1*06:02:18、DQB1*06:02:19、DQB1*06:02:20、DQB1*06:02:21、DQB1*06:02:22、DQB1*06:02:23、DQB1*06:02:24、DQB1*06:02:25、DQB1*06:02:26、DQB1*06:02:27、DQB1*06:02:28、DQB1*06:02:29、DQB1*06:02:30、DQB1*06:02:31、DQB1*06:02:3 2、DQB1*06:02:33、DQB1*06:02:34、DQB1*06:02:35、DQB1*06:02:36、DQB1 *06:02:37、DQB1*06:02:38、DQB1*06:03:01:01、DQB1*06:03:01:02、DQB1* 06:03:01:03、DQB1*06:03:02、DQB1*06:03:03、DQB1*06:03:04、DQB1*06: 03:05、DQB1*06:03:06、DQB1*06:03:07、DQB1*06:03:08、DQB1*06:03:09、D QB1*06:03:10、DQB1*06:03:11、DQB1*06:03:12、DQB1*06:03:13、DQB1*06:03:14、DQB1*06:03:15、DQB1*06:03:16、DQB1*06:03:17、DQB1*06:03:18、DQB1*06:03:19、DQB1*06:03:20、DQB1*06:03:21、DQB1*06:03:22、DQB1*06:03:23、DQB1*06:03:24、DQB1*06:03:25、DQB1*06:03:26、DQB1*06:03:27、DQB1*06:03:28、DQB1*06:03:29、DQB1*06:03:30、DQB1*06:03:31、DQB1*06:03:32、DQB1*06:03:33、DQB1*06:03:34、DQB1*06:03:35、DQB1*06:04:01、DQB1*06:04:02、DQB1*06:04:03、DQB1*06:04:04、DQB1*06:04:05、DQB1*06:04:06、DQB1*06:04:07、DQB1*06:04:08、DQB1*06:04:09、DQB1*06:04: 10、DQB1*06:04:11、DQB1*06:04:12、DQB1*06:05:01、DQB1*06:05:02、DQB1*06:06、DQB1*06:07:01、DQB1*06:07:02、DQB1*06:01、DQB1*06:08:02、DQB1*06:08:03、DQB1*06:09:01:01、DQB1*06:09:01:02、DQB1*06:09:02、DQB1*06:09:03、DQB1*06:09:04、DQB1*06:09:05、DQB1*06:09:06、DQB1*0 6:09:07、DQB1*06:09:08、DQB1*06:09:09、DQB1*06:09:10、DQB1*06:10、DQB1*06:100、DQB1*06:101、DQB1*06:102N、DQB1*06:103、DQB1*06:104、DQB1*06:105、DQB1*06:106、DQB1*06:107、DQB1*06:108、DQB1*06:109、DQB1*06:110、DQB1*06:111、DQB1*06:112N、DQB1*06:113、DQB1*06:114、DQB1*0 6:115、DQB1*06:116、DQB1*06:117、DQB1*06:118:01、DQB1*06:118:02、DQB1*06:118:03、DQB1*06:119、DQB1*06:11:01、DQB1*06:11:02、DQB1*06:11:03、DQB1*06:11:04、DQB1*06:12、DQB1*06:120、DQB1*06:121、DQB1*06:122、DQB1*06:123、DQB1*06:124、DQB1*06:125、DQB1*06:126、DQB1*06:127、DQB1*06:128、DQB1*06:129、DQB1*06:130、DQB1*06:131、DQB1*06:132、DQB1*06:133、DQB1*06:134、DQB1*06:135、DQB1*06:136、DQB1*06:137、DQB1*06:138、DQB1*06:139、DQB1*06:13:01、DQB1*06:13:02、DQB1*06:13:03、DQB1*06:140、DQB1*06:141、DQB1*06:142、DQB1*06:143、DQB1*06:144N、DQ B1*06:145、DQB1*06:146:01、DQB1*06:146:02、DQB1*06:147、DQB1*06:148、DQB1*06:149、DQB1*06:14:01、DQB1*06:14:02、DQB1*06:14:03、DQB1*06:150、DQB1*06:151、DQB1*06:152、DQB1*06:153:01、DQB1*06:153:02、DQB1*06:154、DQB1*06:155、DQB1*06:156、DQB1*06:157、DQB1*06:158N、DQB1* 06:159、DQB1*06:15:01、DQB1*06:15:02、DQB1*06:16、DQB1*06:160、DQB1*06:161、DQB1*06:162、DQB1*06:163、DQB1*06:164、DQB1*06:165、DQB1*06:165 :166、DQB1*06:167、DQB1*06:168、DQB1*06:169、DQB1*06:17、DQB1*06:170、DQB1*06:171、DQB1*06:172、DQB1*06:173、DQB1*06:174、DQB1*06:175、DQB1*06:175 QB1*06:176、DQB1*06:177、DQB1*06:178、DQB1*06:179N、DQB1*06:180、DQB1*06:181、DQB1*06:182、DQB1*06:183、DQB1*06:184、DQB1*06:185、DQB1*06:186、DQB1*06:187、DQB1*06:188、DQB1*06:189、DQB1*06:18:01、DQB1*06:18:02、DQB1*06:190:01、DQB1*06:190:02、DQB1*06:191、DQB1*06:192、DQB1*06:193N、DQB1*06:194、DQB1*06:195、DQB1*06:196、DQB1*06:197、DQB1*06:198、DQB1*06:199、DQB1*06:19:01、DQB1*06:19:02、DQB1*06:20、DQB1*06:200、DQB1*06:201、DQB1*06:202、DQB1*06:203、DQB1*06:204、DQB1*06:205、DQB1*06:206:01、DQB1*06:206:02、DQB1*06:207、DQB1*06:208 ,DQB1*06:209,DQB1*06:21,DQB1*06:210,DQB1*06:211,DQB1*06:212,DQB1*06:213,DQB1*06:214,DQB1*06:215,DQB1*06:216N,DQB1*06:217,DQB1*06:218,DQB1*06:219,DQB1*06:221,DQB1*06:222,DQB1*06:223,DQB1*06:224,DQB1*06:225,DQB1*06:226,DQB1*06:227,DQB1*06:228,DQB1*06:2 29、DQB1*06:22:01、DQB1*06:22:02、DQB1*06:22:03、DQB1*06:23、DQB1*06:230、DQB1*06:231、DQB1*06:232、DQB1*06:233、DQB1*06:234、DQB1*06:235、DQB1*06:236、DQB1*06:237、DQB1*06:238、DQB1*06:239、DQB1*06:24、DQB1*06:240、DQB1*06:241、DQB1*06:242、DQB1*06:243、DQB1*06:244、DQ B1*06:245、DQB1*06:246、DQB1*06:247、DQB1*06:248、DQB1*06:249、DQB1*06:25、DQB1*06:250、DQB1*06:251、DQB1*06:252N、DQB1*06:253、DQB1*06:254、DQB1*06:255、DQB1*06:256、DQB1*06:257、DQB1*06:258、DQB1*06:259、DQB1*06:260、DQB1*06:261、DQB1*06:262、DQB1*06:263、DQB1*06:264、DQB1*06:265、DQB1*06:266、DQB1*06:267、DQB1*06:268、DQB1*06:269、DQB1*06:26N、DQB1*06:270:01、DQB1*06:270:02、DQB1*06:271、DQB1*06:272 、DQB1*06:273、DQB1*06:274、DQB1*06:275、DQB1*06:276、DQB1*06:277、DQB1*06:278、DQB1*06:279、DQB1*06:27:01、DQB1*06:28、DQB1*06:278 QB1*06:280、DQB1*06:281、DQB1*06:282、DQB1*06:283、DQB1*06:284、DQB1*06:285、DQB1*06:286、DQB1*06:287、DQB1*06:288、DQB1*06:289、DQB1*06:29、DQB1*06:290、DQB1*06:291、DQB1*06:292、DQB1*06:293、DQB1*06:294、DQB1*06:295、DQB1*06:296、DQB1*06:297、DQB1*06:298、DQB1*06:299、 DQB1*06:30、DQB1*06:300、DQB1*06:301、DQB1*06:302、DQB1*06:303N、DQB1*06:304N、DQB1*06:305、DQB1*06:306N、DQB1*06:307、DQB1*06:308N、DQB1*06:309、DQB1*06:31、DQB1*06:310、DQB1*06:311、DQB1*06:312、DQB1*06:313、DQB1*06:314、DQB1*06:315、DQB1*06:316、DQB1*06:317N、DQB1*06 :318、DQB1*06:319、DQB1*06:320、DQB1*06:321、DQB1*06:322、DQB1*06:323、DQB1*06:324、DQB1*06:325、DQB1*06:326、DQB1*06:32:01、DQB1*06:32:02、DQB1*06:33、DQB1*06:34、DQB1*06:35、DQB1*06:36、DQB1*06:37、DQB1*06:38、DQB1*06:39、DQB1*06:40、DQB1*06:41、DQB1*06:42、DQB1*06:43、DQB1*06:44, DQB1*06:45, DQB1*06:46, DQB1*06:47, DQB1*06:48:01, DQB1*06:48:02, DQB1*06:49, DQB1*06:50, DQB1*06:51:01, DQB1*06:51:02, DQB1*06:52, DQB1*06:53:01, DQB1*06:53:02, DQB1*06:54N, DQB1*06:55, DQB1*06:56, DQB1*06:57, DQB1*06:58, DQB1*06:59, DQB1*06:60, DQB1*06:61, DQB1*06:62, DQB1*06:63, DQB1*06:64, DQB1*06:65, DQB1*06:66, DQB1*06:67, DQB1*06:68, DQB1*06:69:01, DQB1*06:69:02, DQB1*06:70, DQB1*06:71, DQB1*06:72, DQB1*06:73, DQB1*06:74, DQB1*06:75NX, DQB1*06:76, DQB1*06:77N, DQB1*06:78, DQB1*06:79:01, DQB1*06:79:02, DQB1*06:80, DQB1*06:81, DQB1*06:82, DQB1*06:83, DQB1*06:84, DQB1*06:85, DQB1*06:86, DQB1*06:87, DQB1*06:88, DQB1*06:89, DQB1*06:90, DQB1*06:91, DQB1*06:92:01, DQB1*06:92:02, DQB1*06:93, DQB1*06:94, DQB1*06:95, DQB1*06:96:01, DQB1*06:96:02, DQB1*06:97, DQB1*06:98, DQB1*06:99:01, DQB1*06:99:02 and any combination thereof.

[0138] In some respects, MHC class II molecules comprise a DQβ chain containing an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity with SEQ ID NO:3, wherein the DQβ chain contains tryptophan at position 114 corresponding to amino acid residue 114 of SEQ ID NO:1, and wherein the DQβ chain contains methionine at position 143 corresponding to amino acid residue 143 of SEQ ID NO:1. In some respects, MHC class II molecules comprise a DQβ chain containing an amino acid sequence having at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% sequence identity with SEQ ID NO:3, wherein the DQβ chain comprises (i) tryptophan at position 114 corresponding to amino acid residue 114 of SEQ ID NO:1, (ii) methionine at position 143 corresponding to amino acid residue 143 of SEQ ID NO:1, (iii) glutamine at position 110 corresponding to amino acid residue 110 of SEQ ID NO:1, (iv) valine at position 116 corresponding to amino acid residue 116 of SEQ ID NO:1, (v) histidine at position 118 corresponding to amino acid residue 118 of SEQ ID NO:1, and (vi) glutamine at position 146 corresponding to amino acid residue 146 of SEQ ID NO:1. In some respects, MHC class II molecules contain a DQβ chain, which contains the amino acid sequence shown in SEQ ID NO:3.

[0139] II.A.2. MHC Class II α chain

[0140] In some aspects of this disclosure, MHC class II molecules also comprise an α chain. In some aspects, the α chain is a wild-type α chain. In some aspects, the α chain is a DQα chain. Any DQα chain may be used in the compositions and methods of this disclosure. In some aspects, the DQα chain comprises an HLA-DQA1*01, HLA-DQA1*02, HLA-DQA1*03, HLA-DQA1*04, HLA-DQA1*05, or HLA-DQA1*06 allele. In some aspects, the DQα chain comprises an HLA-DQA1*01 allele. In some aspects, the DQα chain comprises an HLA-DQA1*02 allele. In some aspects, the DQα chain comprises an HLA-DQA1*03 allele. In some aspects, the DQα chain comprises an HLA-DQA1*04 allele. In some respects, the DQα chain contains the HLA-DQA1*05 allele. In other respects, the DQα chain contains the HLA-DQA1*06 allele.

[0141] In some respects, the DQα chain is selected from DQA1*01:01:01:01, DQA1*01:01:01:02, DQA1*01:01:01:03, DQA1*01:01:01:05, DQA1*01:01:01:06, DQA1*01:01:02, and DQA1*01:01:03. , DQA1*01:01:04, DQA1*01:01:05, DQA1*01:02:01:01, DQA1*01:02:01:02, DQA1*01:02:01:03, DQA1*01:02:01:04, DQA1*01:02:01:05, DQA1*01:02: 01:06, DQA1*01:02:01:07, DQA1*01:02:01:08, DQA1*01:02:01:09, DQA1* 01:02:01:10, DQA1*01:02:01:11, DQA1*01:02:01:12, DQA1*01:02:02:01, DQA1*01:02:02:02, DQA1*01:02:02:03, DQA1*01:02:02:04, DQA1*01:02:03, DQA1*01:02:04, DQA1*01:03:01:01, DQA1*01:03:01:02, DQA1*01:03:0 1:03. DQA1*01:03:01:04. DQA1*01:03:01:05. DQA1*01:03:01:06. DQA1*01:03:01:07. DQA1*01:03:01:08. DQA1*01:03:01:09. DQA1*01:04:01:02, DQA1*01:04:01:03, DQA1*01:04:01:04, DQA1*01:04:02, DQA1*01:05:01, DQA1*01:05:02, DQA1*01:06, DQA1*01:07Q, DQA1*01:0 8. DQA1*01:09, DQA1*01:10, DQA1*01:11, DQA1*01:12, DQA1*01:13, DQA1*01:14, DQA1*01:15N, DQA1*01:16N, DQA1*01:17, DQA1*01:18, DQA1*01:19, DQA1*01:20, DQA1*01:21, DQA1*01:22, DQA1*01:23, DQA1*01:24, DQA1*01:25, DQA1*01:26, DQA1*02:01:01:01, DQA1*02:01:01:02, DQA1*02:01:02,DQA1*02:02N、DQA1*02:03、DQA1*03:01:01、DQA1*03:01:03、DQA1*03:02:01:01、DQA1*03:02:01:02、DQA1*03:03:01:01、DQA1*03:03:01:02、DQA1*03:03:01:03、DQA1*03:03:01:04、DQA1*03:03:01:05、DQA1*03:03:01:06、DQA1*03:03:01:07、DQA1*03:03:02、DQA1*03:03:04、DQA1*03:03:05、DQA1*03:03:01:07 06、DQA1*03:07、DQA1*04:01:01:01、DQA1*04:01:01:02、DQA1*04:01:01:03、DQA1*04:01:01:04、DQA1*04:01:01:05、DQA1*04:01:01:06、DQA1*04:01:01:07、DQA1*04:01:01:08、DQA1*04:01:02:01、DQA1*04:01:02:02、DQA1*04:01:03、DQA1*04:02、DQA1*04:03N、DQA1*04:04、DQA1*04:05、DQA1*0 5:01:01:01、DQA1*05:01:01:02、DQA1*05:01:01:03、DQA1*05:01:01:04、DQA1*05:01:02、DQA1*05:01:04、DQA1*05:01:05、DQA1*05:01:06、DQA1*05:02、DQA1*05:03:01:01、DQA1*05:03:01:02、DQA1*05:04、DQA1*05:05:01:01、DQA1*05:05:01:02、DQA1*05:05:01:03、DQA1*05:05:01:04、DQA1*0 5:05:01:05、DQA1*05:05:01:06、DQA1*05:05:01:07、DQA1*05:05:01:08、DQA1*05:05:01:09、DQA1*05:05:01:10、DQA1*05:05:01:11、DQA1*05:05:01:12、DQA1*05:05:01:13、DQA1*05:05:01:14、DQA1*05:05:01:15、DQA1*05:05:01:16、DQA1*05:05:01:17、DQA1*05:05:01:18、DQA1*05:05:01:19、DQA1*05:05:01:20, DQA1*05:06:01:01, DQA1*05:06:01:02, DQA1*05:07, DQA1*05:08, DQA1*05:09, DQA1*05:10, DQA1*05:11, DQA1*05:12, DQA1*05: 13, DQA1*05:14, DQA1*05:15N, DQA1*06:01:01:01, DQA1*06:01:01:02, DQA1*06:01:01:03, DQA1*06:01:01:04, DQA1*06:01:02, DQA1*06:02, and any combination thereof.

[0142] II.A.3 Signal Peptide

[0143] In some aspects, the DQβ chain and / or DQα chain further comprises a signal peptide. Any signal peptide known in the art may be used in the compositions and methods disclosed herein. In some aspects, the DQβ chain signal peptide is the same as the DQα signal peptide. In some aspects, the DQβ chain signal peptide is different from the DQα signal peptide.

[0144] In some respects, the signal peptide is derived from a naturally occurring signal peptide. In some respects, the signal peptide is derived from a naturally occurring DQβ chain signal peptide. In some respects, the signal peptide comprises a naturally occurring DQβ chain signal peptide. In some respects, the signal peptide is derived from a naturally occurring DQα chain signal peptide. In some respects, the signal peptide comprises a naturally occurring DQα chain signal peptide. In some respects, the signal peptide is derived from a fibrin light chain (FibL) signal peptide. In some respects, the signal peptide comprises SEQ ID NO:9. In some respects, the signal peptide is synthetic.

[0145] II.A.4. Transmembrane domain

[0146] In some aspects, the DQβ chain and / or the DQα chain also includes a transmembrane domain. The transmembrane domain can be of any length and of any origin. In some aspects, the length of the transmembrane domain is from at least about 1 to at least about 50 amino acids. In some aspects, the transmembrane domain is derived from a naturally occurring transmembrane domain. In some aspects, the transmembrane domain comprises a naturally occurring transmembrane domain. In some aspects, the transmembrane domain is derived from a naturally occurring HLA transmembrane domain. In some aspects, the transmembrane domain comprises a naturally occurring HLA transmembrane domain. In some aspects, the transmembrane domain is derived from a naturally occurring DQβ chain transmembrane domain. In some aspects, the transmembrane domain comprises a naturally occurring DQβ chain transmembrane domain. In some aspects, the transmembrane domain is derived from a naturally occurring DQα chain transmembrane domain. In some aspects, the transmembrane domain comprises a naturally occurring DQα chain transmembrane domain.

[0147] II.A.5 Leucine Zipper

[0148] In some aspects, the DQβ chain and / or the DQα chain further comprises one or more leucine zipper (LZip) sequences. Any LZip sequence known in the art may be used in the compositions and methods disclosed herein. In some aspects, the DQβ chain and / or the DQα chain comprises an acidic LZip (αLZip), a basic LZip (βLZip), or both. In some aspects, one or more LZip sequences are derived from naturally occurring LZip sequences. In some aspects, one or more LZip sequences comprise naturally occurring LZip sequences. In some aspects, one or more LZip sequences are synthetic. In some aspects, one or more LZip sequences comprise the LZip sequence shown in SEQ ID NO:4 (Table 1).

[0149] II.A.6. Connector

[0150] In some aspects, the DQβ and / or DQα chains used in this disclosure may further comprise a linker. Any linker known in the art may be used in the compositions and methods disclosed herein. In some aspects, the linker comprises a Gly / Ser linker. In some aspects, the linker comprises an amino acid sequence selected from GlySer, Gly2Ser, Gly3Ser, and Gly4Ser. In some aspects, the linker is located at the N-terminus of the extracellular domain of the DQα or DQβ chain. In some aspects, the linker is located at the C-terminus of the extracellular domain of the DQα or DQβ chain. In some aspects, the linker is located between the extracellular domain of the DQα or DQβ chain and a transmembrane domain. In some aspects, the linker is located between the extracellular domain of the DQα or DQβ chain and one or more LZip sequences. In some aspects, the linker is located between the extracellular domain of the DQα or DQβ chain and a signal peptide.

[0151] Linkers of any length can be used in the compositions and methods disclosed herein. In some aspects, the length of the linker is at least one amino acid. In some respects, the length of the linker is at least about 1 to at least about 100, at least about 1 to at least about 90, at least about 1 to at least about 80, at least about 1 to at least about 70, at least about 1 to at least about 60, at least about 1 to at least about 50, at least about 1 to at least about 40, at least about 1 to at least about 30, at least about 1 to at least about 20, at least about 1 to at least about 15, at least about 1 to at least about 14, at least about 1 to at least about 13, at least about 1 to at least about 12, at least about 1 to at least about 11, at least about 1 to at least about 10, at least about 1 to at least about 9, at least about 1 to at least about 8, at least about 1 to at least about 7, at least about 1 to at least about 6, at least about 1 to at least about 5, at least about 1 to at least about 4, at least about 1 to at least about 3 amino acids.

[0152] In some aspects, the length of the linker is at least about 1, at least about 2, at least about 3, at least about 4, at least about 5, at least about 6, at least about 7, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, at least about 13, at least about 14, at least about 15, at least about 20, at least about 30, at least about 40, at least about 50, at least about 60, at least about 70, at least about 80, at least about 90, or at least about 100 amino acids. In some aspects, the length of the linker is about 3 amino acids. In some aspects, the length of the linker is about 4 amino acids. In some aspects, the length of the linker is about 5 amino acids.

[0153] II.B cells

[0154] In some aspects of this disclosure, the MHC class II molecules of this disclosure are attached to or associated with the cell membrane. In some aspects, the β chain of the MHC class II molecules is attached to or associated with the cell membrane. In some aspects, the α chain of the MHC class II molecules is attached to or associated with the cell membrane. In some aspects, both the α and β chains of the MHC class II molecules are attached to or associated with the cell membrane.

[0155] Certain aspects of this disclosure relate to cells comprising the MHC class II molecules disclosed herein. Any cell may be used in the compositions described herein. In some aspects, the cell is a mammalian cell. In some aspects, the cell is an insect cell. In some aspects, the cell is derived from healthy cells, such as healthy fibroblasts. In some aspects, the cell is derived from tumor cells. Non-limiting examples of cells that may be used in this disclosure include K562 cells, T2 cells, HEK293 cells, HEK293T cells, A375 cells, SK-MEL-28 cells, Me275 cells, COS cells, fibroblasts, tumor cells, or any combination thereof. In some aspects, the cell is any cell disclosed in Hasan et al., Adv. Genet. Eng. 4(3):130 (2015) (incorporated herein by reference in its entirety).

[0156] In some respects, the cell is a professional APC. In other respects, the cell is a macrophage, B cell, dendritic cell, or any combination thereof.

[0157] In some respects, the cells lack endogenous expression of one or more MHC class II alleles. In some respects, the cells lack endogenous expression of the HLA-DQ allele. In some respects, the cells lack endogenous expression of the HLA-DQ α chain allele. In some respects, the cells lack endogenous expression of the HLA-DQ β chain allele.

[0158] II.C. Soluble MHC Class II molecules

[0159] In some respects, MHC class II molecules do not associate with cell membranes; for example, MHC class II molecules are in soluble form. As used herein, soluble MHC class II molecules include any MHC class II molecules or portions thereof that do not associate with cell membranes as described herein. In some respects, MHC class II molecules or portions thereof do not bind to any membrane. In some respects, MHC class II molecules or portions thereof bind to inert particles. In some respects, MHC class II molecules or portions thereof bind to the membranes of extracellular vesicles. In some respects, MHC class II molecules bind to artificial membranes or artificial surfaces, such as the surface of an array plate.

[0160] Any inert particles known in the art may be used in the compositions and methods disclosed herein. In some aspects, the inert particles are beads. In some aspects, the beads are glass beads, latex beads, metal beads, or any combination thereof. In some aspects, the inert particles are nanoparticles (NPs). Any NPs known in the art may be used in the compositions and methods disclosed herein. In some aspects, the nanoparticles are selected from polyethylene glycolated iron oxide, chitosan, dextran, gelatin, alginate, liposomes, starch, branched polymers, carbon-based carriers, polylactic acid, poly(cyano)acrylate, polyethyleneimine, block copolymers, polycaprolactone, SPIONs, USPIONs, Cd / Zn-selenide, or silica nanoparticles. In a particular aspect, the nanoparticles are polyethylene glycolated iron oxide nanoparticles. Non-limiting examples of nanoparticles that may be used in the compositions and methods disclosed herein include those set forth in De Jong and Borm, Int. J. Nanomedicine 3(2):133-49 (2008) and Umeshappa et al., Nat. Commun. 10(1):2150 (May 14, 2019) (each incorporated herein by reference in its entirety).

[0161] In some aspects, MHC class II molecules comprise fragments of full-length MHC class II molecules, wherein one or more amino acids of the transmembrane domain of the α chain and / or the transmembrane domain of the β chain are missing. In some aspects, MHC class II molecules comprise extracellular domains of the α chain (e.g., as shown in SEQ ID NO:6) and / or extracellular domains of the β chain (e.g., as shown in SEQ ID NO:1 or 3). In some aspects, MHC class II molecules comprise a DQα chain containing an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with SEQ ID NO:6. In some aspects, MHC class II molecules comprise a DQα chain containing the amino acid sequence shown in SEQ ID NO:6.

[0162] In some aspects, MHC class II molecules comprise a DQβ chain, said DQβ chain comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with SEQ ID NO:1. In some aspects, MHC class II molecules comprise a DQβ chain, said DQβ chain comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with SEQ ID NO:3. In some aspects, MHC class II molecules comprise a DQβ chain, said DQβ chain comprising an amino acid sequence shown in SEQ ID NO:3. In some aspects, MHC class II molecules comprise a DQβ chain, said DQβ chain comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with SEQ ID NO:4. In some aspects, MHC class II molecules comprise a DQβ chain, said DQβ chain comprising the amino acid sequence shown in SEQ ID NO:4. In some aspects, MHC class II molecules comprise a DQβ chain, said DQβ chain comprising an amino acid sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with SEQ ID NO:5. In some aspects, MHC class II molecules comprise a DQβ chain, said DQβ chain comprising the amino acid sequence shown in SEQ ID NO:5.

[0163] II.D. Nucleic Acid Molecules and Vectors

[0164] Certain aspects of this disclosure relate to nucleic acid molecules encoding the MHC class II molecules disclosed herein. In some aspects, the nucleic acid molecules encode the MHC class II β chains disclosed herein. In some aspects, the nucleic acid molecules encoding the MHC class II β chains comprise a nucleotide sequence having at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with the sequence shown in SEQ ID NO:2.

[0165] In some respects, the nucleic acid molecule encodes the MHC class II α chain disclosed herein. In some respects, the nucleic acid molecule encoding the MHC class II α chain comprises a nucleotide sequence having at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% sequence identity with the sequence shown in SEQ ID NO:7.

[0166] In some respects, the nucleic acid molecule encodes both the MHC class II α strand and the MHC class II β strand disclosed herein. In some respects, the sequence encoding the MHC class II α strand is under the control of the same promoter as the sequence encoding the MHC class II β strand. In some respects, the sequence encoding the MHC class II α strand is under the control of a first promoter, and the sequence encoding the MHC class II β strand is under the control of a second promoter.

[0167] In some respects, this disclosure relates to a first nucleic acid molecule encoding the MHC class II β chain disclosed herein and a second nucleic acid molecule encoding the MHC class II α chain disclosed herein.

[0168] Certain aspects of this disclosure relate to a vector or a group of vectors comprising the nucleic acid molecules disclosed herein. In some aspects, the vector is a viral vector. In some aspects, the vector is a viral particle or a virus. In some aspects, the vector is a mammalian vector. In some aspects, the vector is a bacterial vector.

[0169] In some respects, the vector is a retroviral vector. In some respects, the vector is an adenovirus vector, lentivirus, Sendai virus, baculovirus vector, Epstein-Barr virus vector, lactopolyvacuovirus vector, vaccinia virus vector, herpes simplex virus vector, or adeno-associated virus (AAV) vector. In certain respects, the vector is an AAV vector. In some respects, the vector is a lentivirus. In certain respects, the vector is an adenovirus vector. In some respects, the vector is Sendai virus. In some respects, the vector is a hybrid vector. Examples of hybrid vectors that may be used in this disclosure can be found in Huang and Kamihira, Biotechnol. Adv. 31(2):208-23(2103) (incorporated herein by reference in its entirety).

[0170] III. The Method of This Disclosure

[0171] Some aspects of this disclosure relate to methods for treating a disease or condition in a subject. In other aspects, this disclosure relates to methods for enhancing an immune response in a subject in need.

[0172] III.A. Methods of treating tumors

[0173] Certain aspects of this disclosure relate to methods for treating cancer in a subject in need, the methods comprising administering to the subject an HLA class II molecule disclosed herein, a nucleic acid molecule disclosed herein, a vector disclosed herein, or a cell disclosed herein.

[0174] In some respects, cancers include melanoma, bone cancer, kidney cancer, prostate cancer, breast cancer, colon cancer, lung cancer, malignant melanoma of the skin or eye, pancreatic cancer, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, testicular cancer, uterine cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, non-Hodgkin's lymphoma (NHL), primary mediastinal large B-cell lymphoma (PMBC), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), transformed follicular lymphoma, splenic marginal zone lymphoma (SMZL), esophageal cancer, small bowel cancer, endocrine system cancers, thyroid cancer, parathyroid cancer, and kidney cancer. Adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, acute or chronic leukemia, acute myeloid leukemia (AML), chronic myeloid leukemia, acute lymphoblastic leukemia (ALL) (including non-T-cell ALL), chronic lymphocytic leukemia (CLL), childhood solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvis cancer, central nervous system tumors, primary CNS lymphoma, tumor angiogenesis, spinal cord axis tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermoid carcinoma, squamous cell carcinoma, T-cell lymphoma, including those induced by asbestos, environmentally induced cancers, other B-cell malignancies, and any combination of these cancers. In some respects, the cancer is melanoma.

[0175] In some ways, cancer is recurrent. In some ways, cancer is refractory. In some ways, cancer is advanced. In some ways, cancer is metastatic.

[0176] In some respects, the methods disclosed herein treat cancer in subjects. In some respects, the methods disclosed herein reduce the severity of one or more symptoms of cancer. In some respects, the methods disclosed herein reduce the size or number of tumors derived from said cancer. In some respects, the methods disclosed herein increase the overall survival of subjects compared to subjects not provided with the methods disclosed herein. In some respects, the methods disclosed herein increase the progression-free survival of subjects compared to subjects not provided with the methods disclosed herein. In some respects, the methods disclosed herein result in a partial response in subjects. In some respects, the methods disclosed herein result in a complete response in subjects.

[0177] Certain aspects of this disclosure relate to methods for treating infections in subjects in need, said methods comprising administering to the subject an HLA class II molecule disclosed herein, a nucleic acid molecule disclosed herein, a vector disclosed herein, or a cell disclosed herein. Non-limiting examples of infections that can be treated using the compositions and methods disclosed herein include infections caused by viruses (including viroids and prions), bacteria, fungi, parasites, or any combination thereof. In some aspects, the virus is herpesvirus, HIV, papillomavirus, measles virus, rubella virus, human papillomavirus (HPV), human T-lymphovirus I, Epstein-Barr virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, influenza virus, norovirus, and any combination thereof. In some aspects, the bacteria are selected from streptococci, staphylococci, and Escherichia coli. In some embodiments, bacterial infections are selected from brucellosis, Campylobacter infection, cat scratch disease, cholera, Escherichia coli infection, gonorrhea, Klebsiella infection, Enterobacter infection, Serratia infection, Legionella infection, meningococcal infection, pertussis, plague, Pseudomonas infection, Salmonella infection, Shigella infection, typhoid fever, tularemia, anthrax, diphtheria, enterococcal infection, erysipelothricosis, listeriosis, nocardiac infection, pneumococcal infection, staphylococcal infection, streptococcal infection, and any combination thereof. In some embodiments, parasitic infections are selected from pinworm infection, trichomoniasis, toxoplasmosis, giardiasis, cryptosporidiosis, malaria, hookworm infection, tinea, tapeworm infection, trematode infection, and any combination thereof. In some respects, fungal infections are selected from Candida, Malassezia furfur, dermatophytes (e.g., Epidermophyton, Microsporum, and Trichophyton), or any combination thereof.

[0178] In some respects, the methods disclosed herein include treating a subject with cancer or infection, the methods comprising administering to the subject cells described herein, wherein the cells comprise the MHC class II molecules disclosed herein, the nucleic acid molecules disclosed herein, the vectors disclosed herein, or any combination thereof.

[0179] In some cases, the cells are obtained from the subject. In other cases, the cells are obtained from donors other than the subject.

[0180] III.B. Methods for Enriching Target T Cell Populations

[0181] Certain aspects of this disclosure relate to methods for enriching target T cell populations obtained from human subjects. In some aspects, the method includes contacting T cells with HLA class II molecules disclosed herein. In some aspects, the method includes contacting T cells with cells disclosed herein (e.g., APCs). In some aspects, after contact, the enriched T cell population contains a greater number of T cells capable of binding HLA class II molecules compared to the number of T cells capable of binding HLA class II molecules before contact.

[0182] Some aspects of this disclosure relate to methods for selecting T cells capable of targeting diseased cells (e.g., tumor cells). In some aspects, the method includes contacting an isolated population of T cells in vitro with a complex comprising a fragment of an MHC class II molecule and a polypeptide disclosed herein, said polypeptide being, for example, an antigen expressed by diseased cells, such as a tumor-expressed polypeptide, or, for example, an epitope. In some aspects, the T cells are obtained from a human subject.

[0183] T cells obtained from human subjects can be any T cells disclosed herein. In some respects, T cells obtained from human subjects are tumor-infiltrating lymphocytes (TILs).

[0184] In some aspects, the method also includes administering enriched T cells to a human subject. In some aspects, the subject undergoes preconditioning before receiving the T cells as described herein.

[0185] All aspects and options described in this article can be combined in any and all variations.

[0186] All publications, patents and patent applications mentioned in this specification are incorporated herein by reference, as each individual publication, patent or patent application is specifically and individually indicated to the extent of its inclusion herein by reference.

[0187] This disclosure has been generally described, and further understanding can be obtained by referring to the embodiments provided herein. These embodiments are for illustrative purposes only and are not intended to be limiting.

[0188] Example

[0189] Example 1 – Method

[0190] cell

[0191] Peripheral mononuclear cells (Ficoll-Paque PLUS, GE Healthcare LifeSciences, Marlborough, MA) were obtained by density gradient centrifugation. The K562 cell line is an erythroleukemia cell line with defective HLA class I / II expression. A375 is a melanoma cell line. HEK293T cells and A375 cells were grown in DMEM supplemented with 10% FBS and 50 μg / ml gentamicin (Thermo Fisher Scientific, Waltham, MA). K562 and Jurkat 76 cell lines were cultured in RPMI 1640 supplemented with 10% FBS and 50 μg / ml gentamicin.

[0192] peptides

[0193] The synthetic peptides were purchased from Genscript (Piscataway, NJ) and dissolved in DMSO at a concentration of 50 μg / ml.

[0194] Antibody

[0195] The following antibodies were used for flow cytometry analysis: PE-conjugated anti-class II (9-49(I3), Beckman Coulter, Brea, CA; Tü39), APC-Cy7-conjugated anti-CD4 (RPA-T4, Biolegend, San Diego, CA), and PE-conjugated anti-His tag (AD1.1.10, Abcam, Cambridge, MA). Dead cells were distinguished using the Live / Dead Fixable Near-IR Dead Cell Stain Kit 465 (Thermo Fisher Scientific, Waltham, MA). Stained cells were analyzed using Canto II or LSR Tortessa X-20 (BD Biosciences, Franklin Lakes, NJ). Cell sorting was performed using FACS Aria II (BD Biosciences, Franklin Lakes, NJ). Data analysis was performed using FlowJo software (Tree Star, Ashland, OR).

[0196] TCR transduction into primary T cells

[0197] Pan T cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) and CD4 were used respectively. + T-cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) for purifying CD3 + and CD4 + T cells. Purified T cells were stimulated with 200 Gy of irradiated aAPC / mOKT3 at an E:T ratio of 20:1. Starting from the second day, activated T cells were transduced with a cloned TCR gene retrovirus by centrifugation at 1,000 × g for 1 hour at 32°C for 3 consecutive days or using a Retrotronectin-coated plate (Takara Bio, Shiga, Japan). On the second day, 100 IU / ml IL-2 and 10 ng / ml IL-15 were added to the TCR-transduced T cells. The culture medium was replenished every 2–3 days.

[0198] Stain with soluble CD4

[0199] The soluble CD4 (sCD4) gene is generated by fusing the extracellular domain of human CD4 with a 6xHis tag via a GS linker. HEK293T cells were transduced with an sCD4 gene retrovirus, and culture supernatant containing sCD4 monomers was harvested. sCD4 was dimerized with PE-labeled anti-6xHis tag mAbs (AD1.1.10, Abcam, Cambridge, MA) and used. K562 cells expressing HLA class II were stained with dimerized sCD4 for 30 minutes at room temperature in the presence of goat serum. Surface HLA class II expression in K562-derived cells expressing various class II genes is shown below. Figures 3A to 3Q As shown.

[0200] Formation of HLA class II monomers and dimers

[0201] The extracellular domain of the wild-type class II α gene was fused to an acidic leucine zipper via a GGGS linker, followed by fusion to a 6xHis tag via a GS linker (see SEQ ID NO:8). The extracellular domain of the mutant class II β gene (see SEQ ID NO:3) was similarly linked to a basic leucine zipper via a GGGS linker (see SEQ ID NO:4). HEK293T and A375 cells were transfected with the α and β genes using a 293GPG cell-based retroviral system and cultured in DMEM supplemented with 10% FBS and 50 μg / ml gentamicin. DQ5 dimer staining was performed to ensure stable secretion of soluble DQ5. L114W / V143M+4reps HEK293T and A375 cells containing proteins with replacements (N110Q / I116V / S118H / P146N, in addition to L114W / V143M, 4 reps) were grown to confluence, and the culture medium was harvested after 48 hours. In vitro peptide exchange was then performed by mixing the supernatant containing soluble HLA class II with 100 μg / ml of the peptide of interest at 37°C for 20–24 hours. Monomers not subjected to peptide exchange served as controls. Monomer concentrations were measured by specific ELISA using nickel-coated plates (XPressBio, Frederick, MD) and anti-His-tagged biotinylated mAbs (AD1.1.10, R&D Systems, Minneapolis, MN). Soluble HLA class II monomers were dimerized at 4°C for 1.5 hours using PE-conjugated anti-His mAb (AD1.1.10, Abcam, Cambridge, MA) at a 2:1 molar ratio for staining.

[0202] HLA class II dimer staining

[0203] Primary T cells transduced with exogenous TCR gene were incubated at 37°C.46 Pretreated cells with 50 nM dasatinib (LC Laboratories, Woburn, MA) for 30 minutes, and stained with 5-15 μg / ml class II dimers at room temperature for 4-5 hours. After washing, counterstained cell surface molecules with APC-Cy7 conjugated anti-CD4 mAb.

[0204] Statistical analysis

[0205] Statistical analyses were performed using GraphPad Prism 6.0 software (GraphPad Software, San Diego, CA). Unpaired two-tailed Student's t-tests were used for two-sample comparisons. No statistical methods were used to predetermine sample size. Researchers did not blind the allocations during the experiment or outcome evaluation. The experiment was not randomized.

[0206] Example 2 – DQ molecule with enhanced CD4 binding ability

[0207] By introducing the L114W / V143M mutation to generate DQ molecules with enhanced affinity, we determined whether these substitutions could improve the binding of HLA-DQ molecules such as DQ5 (DQA1*01:01-DQB1*05:01) to CD4. In addition to positions 114 and 143, DQB1*05:01 also encodes four different amino acids at positions 110, 116, 118, and 146. Therefore, we generated expression of DQ5... L114W / V143M+4reps K562 cells, except for L114W / V143M ( Figure 1A In addition, it also possesses N110Q / I116V / S118H / P146N substitutions (4 reps), and these cells were stained with sCD4. It expresses DQ5. L114W / V143M+4reps But not expressing DQ5 L114W / V143M DQ5 4reps K562 cells with wild-type DQ5 showed enhanced CD4 binding (or wild-type DQ5 cells showed enhanced CD4 binding). Figures 1B to 1C Importantly, various DQ5 molecules, each expressed at one of the four positions, exhibit a single amino acid inversion. L114W / V143M+4reps A series of K562 cells from mutant strains lacked enhanced CD4 binding ability. Figure 1D These results indicate that the four additional substitutions at N110Q, I116V, S118H, and P146N are crucial for the effectiveness of the L114W / V143M mutation in the observed enhanced DQ5:CD4 binding.

[0208] DQβ chains such as DQB1*02:01, 04:02, and 06:01 encode different amino acids at positions 110, 118, and 146, rather than at position 116. Figure 1E Unlike DQB1*05:01, DQB1*02:01, 04:02, and 06:01 encode Val at position 116, similar to DPB1*04:01, which encodes Val at position 114. All DQ2... L114W / V143M+3reps DQ4 L114W / V143M+3reps and DQ6 L114W / V143M+3reps The mutant (whose β chain carries N110Q, S118H, and P146N substitutions (3 reps) along with L114W / V143M) exhibits enhanced CD4 binding activity. Figure 1F ).

[0209] Example 3 – Affinity-matured DQ dimer specificity and robust staining homologous TCR

[0210] The affinity maturation of DQ dimers carrying the mutations described in Example 2 was evaluated to identify antigen-specific CD4. + The ability of T cells. DQ5 L114W / V143M+4reps and DQ6 L114W / V143M+3reps The dimer successfully stained DQ5-restricted DDX3Y-specific TCR (E6) and DQ6-restricted influenza virus-specific TCR (DM2), respectively. Figures 2A to 2B ).

[0211] To identify class II molecules with mature affinity, this embodiment details several mutations in the β chain rather than the α chain, because the β chain has a more direct interaction with CD4 than the α chain. Further mutations in the α and / or β chains may further enhance the binding between class II and CD4. However, using such soluble class II molecules with excessive CD4 binding capacity may lead to CD4... + Non-specific staining of T cells can have adverse effects.

[0212] In summary, CD4 + T cells play a crucial role in the development of autoimmune diseases and in protection against pathogen infection and cancer. The novel HLA class II multimer technology described in this article can better facilitate the HLA class II-restricted CD4 targeting of the HLA-DQ allele. + Research on T cell responses.

[0213] Example 4

[0214] Wild-type DQ5 and DQ5 L114W / V143M Dimers (Table 3) and DQ5 L114W / V143M+4reps Dimers and their effects on CD4+ transduction of TCR were compared. + T cell staining. Wild-type DQ5 dimer did not detect E6-transduced CD4. + T cells. DQ5L114W / V143M The dimer only shows CD4 transduction to E6. + Weak staining of T cells, compared to DQ5 L114W / V143M+4reps The dimer, on the contrary, showed robust staining. Figures 4A to 4L To verify DQ5 L114W / V143M+4reps Dimer staining, we obtained from dimers amplified in vitro in a peptide-specific manner. + CD4 + GPC3 was cloned in T cells. 138-157 There is a specific DQ5-restricted TCR gene. When in human CD4... + During clonal remodeling in TCR-deficient T cells, the TCR is affected by homologous DQ5. L114W / V143M+4reps The dimer was successfully stained and functioned in a DQ5-restricted and antigen-specific manner. Figures 5A to 5G ).

[0215] Table 3: TCR sequences

[0216]

[0217] method

[0218] cell

[0219] Peripheral mononuclear cells were obtained by density gradient centrifugation. K562-based artificial antigen-presenting cells (aAPCs) expressing various HLA class II genes as single HLA alleles along with CD80 and CD83 have been previously reported (Butler, MO et al., PLoS One 7, e30229 (2012)). The Jurkat 76 cell line is a T-cell leukemia cell line lacking endogenous TCR, CD4, and CD8 expression (see Heemskerk, MH et al., Blood 102, 3530-3540 (2003)). Jurkat 76 / CD4 cells were generated by retroviral transduction of the human CD4 gene. A375 cells are a melanoma cell line. HEK293T cells and A375 cells were grown in DMEM supplemented with 10% FBS and 50 μg / ml gentamicin. The Jurkat 76 cell line was cultured in RPMI 1640 supplemented with 10% FBS and 50 μg / ml gentamicin.

[0220] peptides

[0221] The synthetic peptide was dissolved in DMSO at a concentration of 50 mg / ml.

[0222] Gene

[0223] The novel TCR gene was cloned by rapid 5'-amplification (RACE) PCR of the cDNA ends and sequenced as described above (see, for example, Nakatsugawa, M. et al., Sci Rep 6, 23821 (2016); Nakatsugawa, M. et al., JImmunol 194, 3487-3500 (2015); and Ochi, T. et al., Cancer Immunol Res 3, 1070-1081 (2015); each of which is incorporated herein by reference in its entirety). All genes were cloned into the pMX retroviral vector and transduced into cell lines using retroviral systems based on 293GPG and PG13 cells (see, for example, Hirano, N. et al., Blood 107, 1528-1536 (2006); Butler, Mo et al., Clin Cancer Res 13, 1857-1867 (2007); Hirano, N. et al., Clin Cancer Res 12, 2967-2975 (2006); each reference is incorporated herein by reference in its entirety).

[0224] Antibody

[0225] The following antibodies were used for flow cytometry analysis: APC-Cy7 conjugated anti-CD4 (RPA-T4, BIOLEGEND, San Diego, CA; see Wooldridge, L. et al., Eur J Immunol 36, 1847-1855 (2006)) and PE conjugated anti-His tag (AD1.1.10, ABCAM, Cambridge, MA). Dead cells were distinguished using the LIVE / DEAD Fixable Aqua Dead Cell Stain Kit. Stained cells were analyzed using FACSCanto II or LSR TortessaX-20. Cell sorting was performed using FACSAria II. Data analysis was performed using FlowJo software (version 9.9.6).

[0226] Formation of HLA class II monomers and dimers

[0227] A375 cells were transfected with the α and β genes using a 293GPG-based retroviral system (see, for example, Hirano, N. et al., Blood 107, 1528-1536 (2006); Butler, MO et al., Clin Cancer Res 13, 1857-1867 (2007); and Hirano, N. et al., Blood 108, 2662-2668 (2006); each cited in its entirety). The cells were then cultured in DMEM supplemented with 10% FBS and 50 μg / ml gentamicin. After 48 hours, the conditioned medium was harvested and concentrated by filtration (molecular weight cutoff (MWCO) 10 kDa). In vitro peptide exchange was then performed by mixing the supernatant containing soluble HLA class II peptides with 100 μg / ml of the peptide of interest at 37°C for 20–24 hours. Monomer concentrations were measured by specific ELISA using nickel-coated plates and anti-His-tagged biotinylated mAbs. Soluble HLA class II monomers were dimerized at 4°C for 1.5 hours using PE-conjugated anti-His mAbs at a 2:1 molar ratio for staining.

[0228] DQ5 restriction antigen-specific CD4 + T cell stimulation

[0229] CD4 + T cells were purified and then used with GPC3. 138-157 DQ5-expressing aAPCs were stimulated with a pulse of 10 μg / ml and irradiated at 200 Gy at an E:T ratio of 20:1. 48 hours later, 10 IU / ml IL-2 and 10 ng / ml IL-15 were added to CD4. + In T cells, the culture medium supplemented with IL-2 (10 IU / ml) and IL-15 (10 ng / ml) was replenished every 2-3 days. After two weeks, the T cells were subjected to DQ5 assay. L114W / V143M+4reps Dimer staining.

[0230] HLA class II dimer staining

[0231] Primary CD4 transduced with antigen-specific TCR gene + T cells and Jurkat76 / CD4 T cells were pretreated with 50 nM dasatinib at 37°C for 30 min and stained with class II dimers at 5-15 μg / ml for 4-5 h at room temperature. After washing, cell surface molecules were counterstained with APC-Cy7 conjugated anti-CD4 mAb.

[0232] ELISPOT measurement

[0233] As previously reported, the cytokine ELISPOT assay was performed (see, for example, Yamashita, Y. et al., Nat Commun 8, 15244 (2017); and Anczurowski, M. et al., Sci Rep 8, 4804 (2018); each of which is incorporated herein by reference in its entirety). sequence list <110> University Health Network <120> MHC Class II molecules and their usage <130> 4285.011PC01 / C-K / BMD <150> US 62 / 880,501 <151> 2019-07-30 <150> US 63 / 029,114 <151> 2020-05-22 <160> 15 <170> PatentIn version 3.5 <210> 1 <211> 198 <212> PRT <213> Artificial Sequence <220> <223> Synthetic constructs <400> 1 Arg Asp Ser Pro Glu Asp Phe Val Tyr Gln Phe Lys Gly Leu Cys Tyr 1 5 10 15 Phe Thr Asn Gly Thr Glu Arg Val Arg Gly Val Thr Arg His Ile Tyr 20 25 30 Asn Arg Glu Glu Tyr Val Arg Phe Asp Ser Asp Val Gly Val Tyr Arg 35 40 45 Ala Val Thr Pro Gln Gly Arg Pro Val Ala Glu Tyr Trp Asn Ser Gln 50 55 60 Lys Glu Val Leu Glu Gly Ala Arg Ala Ser Val Asp Arg Val Cys Arg 65 70 75 80 His Asn Tyr Glu Val Ala Tyr Arg Gly Ile Leu Gln Arg Arg Val Glu 85 90 95 Pro Thr Val Thr Ile Ser Pro Ser Arg Thr Glu Ala Leu Asn His His 100 105 110 Asn Leu Leu Ile Cys Ser Val Thr Asp Phe Tyr Pro Ser Gln Ile Lys [[ID=...]] 115 120 125 Val Arg Trp Phe Arg Asn Asp Gln Glu Glu Thr Ala Gly Val Val Ser 130 135 140 Thr Pro Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val Met 145 150 155 160 Leu Glu Met Thr Pro Gln Arg Gly Asp Val Tyr Thr Cys His Val Glu 165 170 175 His Pro Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln Ser 180 185 190 Glu Ser Ala Gln Ser Lys 195 <210> 2 <211> 594 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 2 Ala Gly Ala Gly Ala Cys Thr Cys Thr Cys Cys Cys Gly Ala Gly Gly 1 5 10 15 Ala Thr Thr Thr Cys Gly Thr Gly Thr Ala Cys Cys Ala Gly Thr Thr 20 25 30 Thr Ala Ala Gly Gly Gly Cys Cys Thr Gly Thr Gly Cys Thr Ala Cys 35 40 45 Thr Thr Cys Ala Cys Cys Ala Ala Cys Gly Gly Gly Ala Cys Gly Gly 50 55 60 Ala Gly Cys Gly Cys Gly Thr Gly Cys Gly Gly Gly Gly Thr Gly Thr 65 70 75 80 Gly Ala Cys Cys Ala Gly Ala Cys Ala Cys Ala Thr Cys Thr Ala Thr 85 90 95 Ala Ala Cys Cys Gly Ala Gly Ala Gly Gly Ala Gly Thr Ala Cys Gly 100 105 110 Thr Gly Cys Gly Cys Thr Thr Cys Gly Ala Cys Ala Gly Cys Gly Ala 115 120 125 Cys Gly Thr Gly Gly Gly Gly Gly Thr Gly Thr Ala Cys Cys Gly Gly 130 135 140 Gly Cys Ala Gly Thr Gly Ala Cys Gly Cys Cys Gly Cys Ala Gly Gly 145 150 155 160 Gly Gly Cys Gly Gly Cys Cys Thr Gly Thr Thr Gly Cys Cys Gly Ala 165 170 175 Gly Thr Ala Cys Thr Gly Gly Ala Ala Cys Ala Gly Cys Cys Ala Gly 180 185 190 Ala Ala Gly Gly Ala Ala Gly Thr Cys Cys Thr Gly Gly Ala Gly Gly 195 200 205 Gly Gly Gly Cys Cys Cys Gly Gly Gly Cys Gly Thr Cys Gly Gly Thr 210 215 220 Gly Gly Ala Cys Ala Gly Gly Gly Thr Gly Thr Gly Cys Ala Gly Ala 225 230 235 240 Cys Ala Cys Ala Ala Cys Thr Ala Cys Gly Ala Gly Gly Thr Gly Gly 245 250 255 Cys Gly Thr Ala Cys Cys Gly Cys Gly Gly Gly Ala Thr Cys Cys Thr 260 265 270 Gly Cys Ala Gly Ala Gly Gly Ala Gly Ala Gly Thr Gly Gly Ala Gly 275 280 285 Cys Cys Cys Ala Cys Ala Gly Thr Gly Ala Cys Cys Ala Thr Cys Thr 290 295 300 Cys Cys Cys Cys Ala Thr Cys Cys Ala Gly Gly Ala Cys Ala Gly Ala 305 310 315 320 Gly Gly Cys Cys Cys Thr Cys Ala Ala Cys Cys Ala Cys Cys Ala Cys 325 330 335 Ala Ala Cys Cys Thr Gly Cys Thr Gly Ala Thr Cys Thr Gly Cys Thr 340 345 350 Cys Gly Gly Thr Gly Ala Cys Ala Gly Ala Thr Thr Thr Cys Thr Ala 355 360 365 Thr Cys Cys Ala Ala Gly Cys Cys Ala Gly Ala Thr Cys Ala Ala Ala 370 375 380 Gly Thr Cys Cys Gly Gly Thr Gly Gly Thr Thr Thr Cys Gly Gly Ala 385 390 395 400 Ala Thr Gly Ala Thr Cys Ala Gly Gly Ala Gly Gly Ala Gly Ala Cys 405 410 415 Ala Gly Cys Cys Gly Gly Cys Gly Thr Thr Gly Thr Gly Thr Cys Cys 420 425 430 Ala Cys Cys Cys Cys Cys Cys Thr Cys Ala Thr Thr Ala Gly Gly Ala 435 440 445 Ala Cys Gly Gly Thr Gly Ala Cys Thr Gly Gly Ala Cys Cys Thr Thr 450 455 460 Cys Cys Ala Gly Ala Thr Cys Cys Thr Gly Gly Thr Gly Ala Thr Gly 465 470 475 480 Cys Thr Gly Gly Ala Ala Ala Thr Gly Ala Cys Thr Cys Cys Cys Cys 485 490 495 Ala Gly Cys Gly Thr Gly Gly Ala Gly Ala Thr Gly Thr Cys Thr Ala 500 505 510 Cys Ala Cys Cys Thr Gly Cys Cys Ala Cys Gly Thr Gly Gly Ala Gly 515 520 525 Cys Ala Cys Cys Cys Cys Ala Gly Cys Cys Thr Cys Cys Ala Gly Ala 530 535 540 Gly Cys Cys Cys Cys Ala Thr Cys Ala Cys Cys Gly Thr Gly Gly Ala 545 550 555 560 Gly Thr Gly Gly Cys Gly Gly Gly Cys Thr Cys Ala Gly Thr Cys Thr 565 570 575 Gly Ala Ala Thr Cys Thr Gly Cys Cys Cys Ala Gly Ala Gly Cys Ala 580 585 590 Ala Gly [[ID=_{32}]] <210> 3 <211> 198 <21^2> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 3 Arg Asp Ser Pro Glu Asp Phe Val Tyr Gln Phe Lys Gly Leu Cys Tyr 1 5 10 15 Phe Thr Asn Gly Thr Glu Arg Val Arg Gly Val Thr Arg His Ile Tyr 20 25 30 Asn Arg Glu Glu Tyr Val Arg Phe Asp Ser Asp Val Gly Val Tyr Arg 35 40 45 Ala Val Thr Pro Gln Gly Arg Pro Val Ala Glu Tyr Trp Asn Ser Gln 50 55 60 Lys Glu Val Leu Glu Gly Ala Arg Ala Ser Val Asp Arg Val Cys Arg 65 70 75 80 His Asn Tyr Glu Val Ala Tyr Arg Gly Ile Leu Gln Arg Arg Val Glu 85 90 95 Pro Thr Val Thr Ile Ser Pro Ser Arg Thr Glu Ala Leu Gln His His 100 105 110 Asn Trp Leu Val Cys His Val Thr Asp Phe Tyr Pro Ser Gln Ile Lys 115 120 125 Val Arg Trp Phe Arg Asn Asp Gln Glu Glu Thr Ala Gly Val Met Ser 130 135 140 Thr Asn Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val Met 145 150 155 160 Leu Glu Met Thr Pro Gln Arg Gly Asp Val Tyr Thr Cys His Val Glu 165 170 175 His Pro Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln Ser 180 185 190 Glu Ser Ala Gln Ser Lys 195 <210> 4 <211> 267 <212> PRT <213> Artificial Sequence(Artificial Sequence) <220> <223> synthetic construction <400> 4 Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala Arg Asp Ser Pro Glu Asp Phe Val Tyr Gln Phe Lys Gly Leu Cys 20 25 30 Tyr Phe Thr Asn Gly Thr Glu Arg Val Arg Gly Val Thr Arg His Ile 35 40 45 Tyr Asn Arg Glu Glu Tyr Val Arg Phe Asp Ser Asp Val Gly Val Tyr 50 55 60 Arg Ala Val Thr Pro Gln Gly Arg Pro Val Ala Glu Tyr Trp Asn Ser 65 70 75 80 Gln Lys Glu Val Leu Glu Gly Ala Arg Ala Ser Val Asp Arg Val Cys 85 90 95 Arg His Asn Tyr Glu Val Ala Tyr Arg Gly Ile Leu Gln Arg Arg Val 100 105 110 Glu Pro Thr Val Thr Ile Ser Pro Ser Arg Thr Glu Ala Leu Gln His 115 120 125 His Asn Trp Leu Val Cys His Val Thr Asp Phe Tyr Pro Ser Gln Ile 130 135 140 Lys Val Arg Trp Phe Arg Asn Asp Gln Glu Glu Thr Ala Gly Val Met 145 150 155 160 Ser Thr Asn Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val 165 170 175 Met Leu Glu Met Thr Pro Gln Arg Gly Asp Val Tyr Thr Cys His Val 180 185 190 Glu His Pro Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln 195 200 205 Ser Glu Ser Ala Gln Ser Lys Gly Gly Gly Gly Ser Leu Glu Ile Glu 210 215 220 Ala Ala Phe Leu Glu Arg Glu Asn Thr Ala Leu Glu Thr Arg Val Ala 225 230 235 240 Glu Leu Arg Gln Arg Val Gln Arg Leu Arg Asn Arg Val Ser Gln Tyr 245 250 255 Arg Thr Arg Tyr Gly Pro Leu Gly Gly Gly Lys 260 265 <210> 5 <211> 261 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 5 Met Ser Trp Lys Lys Ser Leu Arg Ile Pro Gly Asp Leu Arg Val Ala 1 5 10 15 Thr Val Thr Leu Met Leu Ala Ile Leu Ser Ser Ser Leu Ala Glu Gly 20 25 30 Arg Asp Ser Pro Glu Asp Phe Val Tyr Gln Phe Lys Gly Leu Cys Tyr 35 40 45 Phe Thr Asn Gly Thr Glu Arg Val Arg Gly Val Thr Arg His Ile Tyr 50 55 60 Asn Arg Glu Glu Tyr Val Arg Phe Asp Ser Asp Val Gly Val Tyr Arg 65 70 75 80 Ala Val Thr Pro Gln Gly Arg Pro Val Ala Glu Tyr Trp Asn Ser Gln 85 90 95 Lys Glu Val Leu Glu Gly Ala Arg Ala Ser Val Asp Arg Val Cys Arg 100 105 110 His Asn Tyr Glu Val Ala Tyr Arg Gly Ile Leu Gln Arg Arg Val Glu 115 120 125 Pro Thr Val Thr Ile Ser Pro Ser Arg Thr Glu Ala Leu Asn His His 130 135 140 Asn Leu Leu Ile Cys Ser Val Thr Asp Phe Tyr Pro Ser Gln Ile Lys 145 150 155 160 Val Arg Trp Phe Arg Asn Asp Gln Glu Glu Thr Ala Gly Val Val Ser 165 170 175 Thr Pro Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val Met 180 185 190 Leu Glu Met Thr Pro Gln Arg Gly Asp Val Tyr Thr Cys His Val Glu 195 200 205 His Pro Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln Ser 210 215 220 Glu Ser Ala Gln Ser Lys Met Leu Ser Gly Val Gly Gly Phe Val Leu 225 230 235 240 Gly Leu Ile Phe Leu Gly Leu Gly Leu Ile Ile Arg Gln Arg Ser Arg 245 250 255 Lys Gly Leu Leu His 260 <210> 6 <211> 195 <212> PRT <213> Artificial Sequence[[ID=4x]] <220> <223> Synthetic construct <400> 6 Glu Asp Ile Val Ala Asp His Val Ala Ser Cys Gly Val Asn Leu Tyr 1 5 10 15 Gln Phe Tyr Gly Pro Ser Gly Gln Tyr Thr His Glu Phe Asp Gly Asp 20 25 30 Glu Glu Phe Tyr Val Asp Leu Glu Arg Lys Glu Thr Ala Trp Arg Trp 35 40 45 Pro Glu Phe Ser Lys Phe Gly Gly Phe Asp Pro Gln Gly Ala Leu Arg 50 55 60 Asn Met Ala Val Ala Lys His Asn Leu Asn Ile Met Ile Lys Arg Tyr 65 70 75 80 Asn Ser Thr Ala Ala Thr Asn Glu Val Pro Glu Val Thr Val Phe Ser 85 90 95 Lys Ser Pro Val Thr Leu Gly Gln Pro Asn Thr Leu Ile Cys Leu Val 100 105 110 Asp Asn Ile Phe Pro Pro Val Val Asn Ile Thr Trp Leu Ser Asn Gly 115 120 125 Gln Ser Val Thr Glu Gly Val Ser Glu Thr Ser Phe Leu Ser Lys Ser 130 135 140 Asp His Ser Phe Phe Lys Ile Ser Tyr Leu Thr Phe Leu Pro Ser Ala 145 150 155 160 Asp Glu Ile Tyr Asp Cys Lys Val Glu His Trp Gly Leu Asp Gln Pro 165 170 175 Leu Leu Lys His Trp Glu Pro Glu Ile Pro Ala Pro Met Ser Glu Leu 180 185 190 Thr Glu Thr 195 <210> 7 <211> 585 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 7 Gly Ala Gly Gly Ala Cys Ala Thr Cys Gly Thr Gly Gly Cys Cys Gly 1 5 10 15 Ala Thr Cys Ala Cys Gly Thr Gly Gly Cys Ala Ala Gly Cys Thr Gly 20 25 30 Cys Gly Gly Cys Gly Thr Gly Ala Ala Cys Cys Thr Gly Thr Ala Cys 35 40 45 Cys Ala Gly Thr Thr Cys Thr Ala Cys Gly Gly Cys Cys Cys Cys Thr 50 55 60 Cys Thr Gly Gly Cys Cys Ala Gly Thr Ala Cys Ala Cys Cys Cys Ala 65 70 75 80 Thr Gly Ala Ala Thr Thr Thr Gly Ala Thr Gly Gly Ala Gly Ala Thr 85 90 95 Gly Ala Gly Gly Ala Gly Thr Thr Cys Thr Ala Cys Gly Thr Gly Gly 100 105 110 Ala Cys Cys Thr Gly Gly Ala Gly Ala Gly Gly Ala Ala Gly Gly Ala 115 120 125 Gly Ala Cys Thr Gly Cys Cys Thr Gly Gly Cys Gly Gly Thr Gly Gly 130 135 140 Cys Cys Thr Gly Ala Gly Thr Thr Cys Ala Gly Cys Ala Ala Ala Thr 145 150 155 160 Thr Thr Gly Gly Ala Gly Gly Thr Thr Thr Thr Gly Ala Cys Cys Cys 165 170 175 Gly Cys Ala Gly Gly Gly Thr Gly Cys Ala Cys Thr Gly Ala Gly Ala 180 185 190 Ala Ala Cys Ala Thr Gly Gly Cys Thr Gly Thr Gly Gly Cys Ala Ala 195 200 205 Ala Ala Cys Ala Cys Ala Ala Cys Thr Thr Gly Ala Ala Cys Ala Thr 210 215 220 Cys Ala Thr Gly Ala Thr Thr Ala Ala Ala Cys Gly Cys Thr Ala Cys 225 230 235 240 Ala Ala Cys Thr Cys Thr Ala Cys Cys Gly Cys Thr Gly Cys Thr Ala 245 250 255 Cys Cys Ala Ala Thr Gly Ala Gly Gly Thr Thr Cys Cys Thr Gly Ala 260 265 270 Gly Gly Thr Cys Ala Cys Ala Gly Thr Gly Thr Thr Thr Thr Cys Cys 275 280 285 Ala Ala Gly Thr Cys Thr Cys Cys Cys Gly Thr Gly Ala Cys Ala Cys 290 295 300 Thr Gly Gly Gly Thr Cys Ala Gly Cys Cys Cys Ala Ala Cys Ala Cys 305 310 315 320 Cys Cys Thr Cys Ala Thr Thr Thr Gly Thr Cys Thr Thr Gly Thr Gly 325 330 335 Gly Ala Cys Ala Ala Cys Ala Thr Cys Thr Thr Thr Cys Cys Thr Cys 340 345 350 Cys Thr Gly Thr Gly Gly Thr Cys Ala Ala Cys Ala Thr Cys Ala Cys 355 360 365 Ala Thr Gly Gly Cys Thr Gly Ala Gly Cys Ala Ala Thr Gly Gly Gly 370 375 380 Cys Ala Gly Thr Cys Ala Gly Thr Cys Ala Cys Ala Gly Ala Ala Gly 385 390 395 400 Gly Thr Gly Thr Thr Thr Cys Thr Gly Ala Gly Ala Cys Cys Ala Gly 405 410 415 Cys Thr Thr Cys Cys Thr Cys Thr Cys Cys Ala Ala Gly Ala Gly Thr 420 425 430 Gly Ala Thr Cys Ala Thr Thr Cys Cys Thr Thr Cys Thr Thr Cys Ala 435 440 445 Ala Gly Ala Thr Cys Ala Gly Thr Thr Ala Cys Cys Thr Cys Ala Cys 450 455 460 Cys Thr Thr Cys Cys Thr Cys Cys Cys Thr Thr Cys Thr Gly Cys Thr 465 470 475 480 Gly Ala Thr Gly Ala Gly Ala Thr Thr Thr Ala Thr Gly Ala Cys Thr 485 490 495 Gly Cys Ala Ala Gly Gly Thr Gly Gly Ala Gly Cys Ala Cys Thr Gly 500 505 510 Gly Gly Gly Cys Cys Thr Gly Gly Ala Cys Cys Ala Gly Cys Cys Thr 515 520 525 Cys Thr Thr Cys Thr Gly Ala Ala Ala Cys Ala Cys Thr Gly Gly Gly 530 535 540 Ala Gly Cys Cys Thr Gly Ala Gly Ala Thr Thr Cys Cys Ala Gly Cys 545 550 555 560 Cys Cys Cys Thr Ala Thr Gly Thr Cys Ala Gly Ala Gly Cys Thr Cys 565 570 575 Ala Cys Ala Gly Ala Gly Ala Cys Thr 580 585 <210> 8 <211> 272 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 8 Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15​​​​​​​​​​​​​​​​​​​​​​ 100 105 110 Ser Lys Ser Pro Val Thr Leu Gly Gln Pro Asn Thr Leu Ile Cys Leu 115 120 125 Val Asp Asn Ile Phe Pro Pro Val Val Asn Ile Thr Trp Leu Ser Asn 130 135 140 Gly Gln Ser Val Thr Glu Gly Val Ser Glu Thr Ser Phe Leu Ser Lys 145 150 155 160 Ser Asp His Ser Phe Phe Lys Ile Ser Tyr Leu Thr Phe Leu Pro Ser 165 170 175 Ala Asp Glu Ile Tyr Asp Cys Lys Val Glu His Trp Gly Leu Asp Gln 180 185 190 Pro Leu Leu Lys His Trp Glu Pro Glu Ile Pro Ala Pro Met Ser Glu 195 200 205 Leu Thr Glu Thr Gly Gly Gly Gly Ser Leu Glu Ile Arg Ala Ala Phe 210 215 220 Leu Arg Gln Arg Asn Thr Ala Leu Arg Thr Glu Val Ala Glu Leu Glu 225 230 235 240 Gln Glu Val Gln Arg Leu Glu Asn Glu Val Ser Gln Tyr Glu Thr Arg 245 250 255 Tyr Gly Pro Leu Gly Gly Gly Lys Gly Ser His His His His His 260 265 270 <210> 9 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Synthetic constructs <400> 9 Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala <210> 10 <211> 458 <212> PRT <213> Artificial Sequence <220> <223> Synthetic constructs <400> 10 Met Asn Arg Gly Val Pro Phe Arg His Leu Leu Leu Val Leu Gln Leu 1 5 10 15 Ala Leu Leu Pro Ala Ala Thr Gln Gly Lys Lys Val Val Leu Gly Lys 20 25 30 Lys Gly Asp Thr Val Glu Leu Thr Cys Thr Ala Ser Gln Lys Lys Ser 35 40 45 Ile Gln Phe His Trp Lys Asn Ser Asn Gln Ile Lys Ile Leu Gly Asn 50 55 60 Gln Gly Ser Phe Leu Thr Lys Gly Pro Ser Lys Leu Asn Asp Arg Ala 65 70 75 80 Asp Ser Arg Arg Ser Leu Trp Asp Gln Gly Asn Phe Pro Leu Ile Ile 85 90 95 Lys Asn Leu Lys Ile Glu Asp Ser Asp Thr Tyr Ile Cys Glu Val Glu 100 105 110 Asp Gln Lys Glu Glu Val Gln Leu Leu Val Phe Gly Leu Thr Ala Asn 115 120 125 Ser Asp Thr His Leu Leu Gln Gly Gln Ser Leu Thr Leu Thr Leu Glu 130 135 140 Ser Pro Pro Gly Ser Ser Pro Ser Val Gln Cys Arg Ser Pro Arg Gly 145 150 155 160 Lys Asn Ile Gln Gly Gly Lys Thr Leu Ser Val Ser Gln Leu Glu Leu 165 170 175 Gln Asp Ser Gly Thr Trp Thr Cys Thr Val Leu Gln Asn Gln Lys Lys 180 185 190 Val Glu Phe Lys Ile Asp Ile Val Val Leu Ala Phe Gln Lys Ala Ser 195 200 205 Ser Ile Val Tyr Lys Lys Glu Gly Glu Gln Val Glu Phe Ser Phe Pro 210 215 220 Leu Ala Phe Thr Val Glu Lys Leu Thr Gly Ser Gly Glu Leu Trp Trp 225 230 235 240 Gln Ala Glu Arg Ala Ser Ser Ser Lys Ser Trp Ile Thr Phe Asp Leu 245 250 255 Lys Asn Lys Glu Val Ser Val Lys Arg Val Thr Gln Asp Pro Lys Leu 260 265 270 Gln Met Gly Lys Lys Leu Pro Leu His Leu Thr Leu Pro Gln Ala Leu 275 280 285 Pro Gln Tyr Ala Gly Ser Gly Asn Leu Thr Leu Ala Leu Glu Ala Lys 290 295 300 Thr Gly Lys Leu His Gln Glu Val Asn Leu Val Val Met Arg Ala Thr 305 310 315 320 Gln Leu Gln Lys Asn Leu Thr Cys Glu Val Trp Gly Pro Thr Ser Pro 325 330 335 Lys Leu Met Leu Ser Leu Lys Leu Glu Asn Lys Glu Ala Lys Val Ser 340 345 350 Lys Arg Glu Lys Ala Val Trp Val Leu Asn Pro Glu Ala Gly Met Trp 355 360 365 Gln Cys Leu Leu Ser Asp Ser Gly Gln Val Leu Leu Glu Ser Asn Ile 370 375 380 Lys Val Leu Pro Thr Trp Ser Thr Pro Val Gln Pro Met Ala Leu Ile 385 390 395 400 Val Leu Gly Gly Val Ala Gly Leu Leu Leu Phe Ile Gly Leu Gly Ile 405 410 415 Phe Phe Cys Val Arg Cys Arg His Arg Arg Arg Gln Ala Glu Arg Met 420 425 430 Ser Gln Ile Lys Arg Leu Leu Ser Glu Lys Lys Thr Cys Gln Cys Pro 435 440 445 His Arg Phe Gln Lys Thr Cys Ser Pro Ile 450 455 <210> 11 <211> 198 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 11 Arg Asp Ser Pro Glu Asp Phe Val Tyr Gln Phe Lys Gly Leu Cys Tyr 1 5 10 15 Phe Thr Asn Gly Thr Glu Arg Val Arg Gly Val Thr Arg His Ile Tyr 20 25 30 Asn Arg Glu Glu Tyr Val Arg Phe Asp Ser Asp Val Gly Val Tyr Arg 35 40 45 Ala Val Thr Pro Gln Gly Arg Pro Val Ala Glu Tyr Trp Asn Ser Gln 50 55 60 Lys Glu Val Leu Glu Gly Ala Arg Ala Ser Val Asp Arg Val Cys Arg 65 70 75 80 His Asn Tyr Glu Val Ala Tyr Arg Gly Ile Leu Gln Arg Arg Val Glu 85 90 95 Pro Thr Val Thr Ile Ser Pro Ser Arg Thr Glu Ala Leu Asn His His 100 105 110 Asn Trp Leu Ile Cys Ser Val Thr Asp Phe Tyr Pro Ser Gln Ile Lys 115 120 125 Val Arg Trp Phe Arg Asn Asp Gln Glu Glu Thr Ala Gly Val Met Ser 130 135 140 Thr Pro Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val Met 145 150 155 160 Leu Glu Met Thr Pro Gln Arg Gly Asp Val Tyr Thr Cys His Val Glu 165 170 175 His Pro Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln Ser 180 185 190 Glu Ser Ala Gln Ser Lys 195 <210> 12 <211> 267 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 12 Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala Arg Asp Ser Pro Glu Asp Phe Val Tyr Gln Phe Lys Gly Leu Cys 20 25 30 Tyr Phe Thr Asn Gly Thr Glu Arg Val Arg Gly Val Thr Arg His Ile 35 40 45 Tyr Asn Arg Glu Glu Tyr Val Arg Phe Asp Ser Asp Val Gly Val Tyr 50 55 60 Arg Ala Val Thr Pro Gln Gly Arg Pro Val Ala Glu Tyr Trp Asn Ser 65 70 75 80 Gln Lys Glu Val Leu Glu Gly Ala Arg Ala Ser Val Asp Arg Val Cys 85 90 95 Arg His Asn Tyr Glu Val Ala Tyr Arg Gly Ile Leu Gln Arg Arg Val 100 105 110 Glu Pro Thr Val Thr Ile Ser Pro Ser Arg Thr Glu Ala Leu Asn His 115 120 125 His Asn Leu Leu Ile Cys Ser Val Thr Asp Phe Tyr Pro Ser Gln Ile 130 135 140 Lys Val Arg Trp Phe Arg Asn Asp Gln Glu Glu Thr Ala Gly Val Val 145 150 155 160 Ser Thr Pro Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val 165 170 175 Met Leu Glu Met Thr Pro Gln Arg Gly Asp Val Tyr Thr Cys His Val 180 185 190 Glu His Pro Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln 195 200 205 Ser Glu Ser Ala Gln Ser Lys Gly Gly Gly Gly Ser Leu Glu Ile Glu 210 215 220 Ala Ala Phe Leu Glu Arg Glu Asn Thr Ala Leu Glu Thr Arg Val Ala 225 230 235 240 Glu Leu Arg Gln Arg Val Gln Arg Leu Arg Asn Arg Val Ser Gln Tyr 245 250 255 Arg Thr Arg Tyr Gly Pro Leu Gly Gly Gly Lys 260 265 <210> 13 <211> 267<00011​​​​​​​​​​Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala Arg Asp Ser Pro Glu Asp Phe Val Tyr Gln Phe Lys Gly Leu Cys 20 25 30 Tyr Phe Thr Asn Gly Thr Glu Arg Val Arg Gly Val Thr Arg His Ile 35 40 45 Tyr Asn Arg Glu Glu Tyr Val Arg Phe Asp Ser Asp Val Gly Val Tyr 50 55 60 Arg Ala Val Thr Pro Gln Gly Arg Pro Val Ala Glu Tyr Trp Asn Ser 65 70 75 80 Gln Lys Glu Val Leu Glu Gly Ala Arg Ala Ser Val Asp Arg Val Cys 85 90 95 Arg His Asn Tyr Glu Val Ala Tyr Arg Gly Ile Leu Gln Arg Arg Val 100 105 110 Glu Pro Thr Val Thr Ile Ser Pro Ser Arg Thr Glu Ala Leu Asn His 115 120 125 His Asn Trp Leu Ile Cys Ser Val Thr Asp Phe Tyr Pro Ser Gln Ile 130 135 140 Lys Val Arg Trp Phe Arg Asn Asp Gln Glu Glu Thr Ala Gly Val Met 145 150 155 160 Ser Thr Pro Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val 165 170 175 Met Leu Glu Met Thr Pro Gln Arg Gly Asp Val Tyr Thr Cys His Val 180 185 190 Glu His Pro Ser Leu Gln Ser Pro Ile Thr Val Glu Trp Arg Ala Gln 195 200 205 Ser Glu Ser Ala Gln Ser Lys Gly Gly Gly Gly Ser Leu Glu Ile Glu 210 215 220 Ala Ala Phe Leu Glu Arg Glu Asn Thr Ala Leu Glu Thr Arg Val Ala 225 230 235 240 Glu Leu Arg Gln Arg Val Gln Arg Leu Arg Asn Arg Val Ser Gln Tyr 245 250 255 Arg Thr Arg Tyr Gly Pro Leu Gly Gly Gly Lys 260 265 <210> 14 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 14 Cys Ala Leu Tyr Thr Asn Ala Gly Lys Ser Thr Phe 1 5 10 <210> 15 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic constructs <400> 15 Cys Ala Thr Ser Arg Asp Val Ser Ser Thr Asp Thr Gln Tyr Phe 1 5 10 15

Claims

1. An HLA class II molecule comprising a DQ β chain, wherein the DQ β chain comprises an extracellular domain, wherein the amino acid sequence of the extracellular domain of the DQ β chain is as shown in SEQ ID NO: 3, and The β chain of HLA class II molecules contains the HLA-DQB1*05:01 allele.

2. The HLA class II molecule of claim 1, wherein the DQ β chain comprises the amino acid sequence shown in SEQ ID NO:

4.

3. The HLA class II molecule as described in claim 1 or 2, further comprising a DQ α chain.

4. The HLA class II molecule of claim 3, wherein the DQ α chain comprises the amino acid sequence shown in SEQ ID NO: 6 or 8.

5. The HLA class II molecule according to any one of claims 1 to 4, wherein the DQ β chain has increased affinity for CD4 protein compared to a reference HLA class II molecule, wherein the DQ β chain comprising the reference HLA class II molecule comprises (i) leucine at position 114 of amino acid residue SEQ ID NO: 1 and / or (ii) valine at position 143 of amino acid residue SEQ ID NO:

1.

6. The HLA class II molecule as claimed in any one of claims 1 to 5, wherein the DQ β chain binds to the cell membrane.

7. The HLA class II molecule as claimed in any one of claims 1 to 6, wherein the DQ β chain comprises an extracellular domain of the full-length DQ β chain.

8. The HLA class II molecule as claimed in any one of claims 3 to 7, wherein the DQ α chain binds to the cell membrane.

9. The HLA class II molecule as claimed in any one of claims 3 to 8, wherein the DQ α chain comprises an extracellular domain of the full-length DQ α chain.

10. The HLA class II molecule according to any one of claims 1 to 9, wherein the DQ β chain comprises a signal peptide.

Images