MHC class ii molecules and methods of using the same

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 DP beta chain having: (i) an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) an amino acid other than valine at a position corresponding to amino acid residue 141 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 / 880,496, filed July 30, 2019, and U.S. Provisional Application No. 63 / 029,111, filed May 22, 2020, each of which is incorporated by reference in its entirety.

[0003] Incorporation by Reference of Sequence Listing Submitted Electronically via EFS-WEB

[0004] The content of the electronically submitted sequence listing (name: 4285.010PC02_SL_ST25.txt; size: 144,830 bytes; date created: July 28, 2020) is incorporated herein by reference in its entirety. SUMMARY

[0005] The present disclosure provides major histocompatibility complex (MHC) class II molecules with increased affinity for CD4 and uses thereof. SUMMARY

[0007] Immunotherapy has become a key tool against a variety of diseases, including cancer. T cell therapy is at the forefront of immunotherapy development, and the adoptive transfer of anti-tumor T cells has been shown to induce clinical responses in cancer patients.

[0008] Directed T cell therapy using T cells expressing T cell receptors (TCRs) specific for target epitopes expressed by tumor cells is a promising form of T cell therapy. Antigen presenting cells display peptide fragments associated with major histocompatibility complex (MHC) on their surface to induce an immune response. Improved presentation of endogenous peptides by class II has been shown to correlate with improved survival rates in cancer patients. However, the development of new TCRs capable of specifically targeting MHC class II presented peptides is hindered by the low affinity of MHC class II proteins for CD4 expressed by T cells.

[0009] The present disclosure provides MHC class II proteins with increased affinity for CD4 and methods of using them to identify and develop new MHC class II specific TCRs. SUMMARY

[0010] Certain aspects of the present disclosure relate to HLA class II molecules comprising a DP beta chain, wherein the DP beta chain comprises an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1.

[0011] Certain aspects of the present disclosure relate to an HLA class II molecule comprising a DP b chain, wherein the DP b chain comprises a substitution mutation at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, wherein the substitution mutation is a substitution with an amino acid other than leucine.

[0012] In some aspects, the DP b chain further comprises an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1.

[0013] Certain aspects of the present disclosure relate to an HLA class II molecule comprising a DP b chain, wherein the DP b chain comprises an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1.

[0014] Certain aspects of the present disclosure relate to an HLA class II molecule comprising a DP b chain, wherein the DP b chain comprises a substitution mutation at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, wherein the substitution mutation is a substitution with an amino acid other than valine.

[0015] In some aspects, the DP b chain further comprises an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1.

[0016] In some aspects, the DP b 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 to an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 4, and 5.

[0017] In some aspects, the amino acid other than leucine comprises a hydrophobic side chain.

[0018] In some aspects, the amino acid other than leucine is selected from the group consisting of alanine, valine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan.

[0019] In some aspects, the amino acid other than leucine is tryptophan.

[0020] In some aspects, the amino acid other than valine comprises a hydrophobic side chain. In some aspects, the amino acid other than valine is selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, the amino acid other than valine is methionine.

[0021] In some aspects, the DP beta chain comprises a tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, and a methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1.

[0022] In some aspects, the DP beta chain comprises an amino acid sequence set forth in SEQ ID NO: 3. In some aspects, the DP beta chain comprises an amino acid sequence set forth in SEQ ID NO: 4.

[0023] DPB1*194, DPB1*195, DPB1*196, DPB1*197, DPB1*198, DPB1*199, DPB1*20, DPB1*200, DPB1*201, DPB1*202, DPB1*203, DPB1*204, DPB1*205, DPB1*206, DPB1*207, DPB1*208, DPB1*209, DPB1*21, DPB1*210, DPB1*211, DPB1*212, DPB1*213, DPB1*214, DPB1*215, DPB1*216, DPB1*217, DPB1*218, DPB1*219, DPB1*22, DPB1*220, DPB1*221, DPB1*222, DPB1*223, DPB1*224, DPB1*225, DPB1*226, DPB1*227, DPB1*228, DPB1*229, DPB1*23, DPB1*230, DPB1*231, DPB1*232, DPB1*233, DPB1*234, DPB1*235, DPB1*236, DPB1*237, DPB1*238, DPB1*239, DPB1*24, DPB1*240, DPB1*241, DPB1*242, DPB1*243, DPB1*244, DPB1*245, DPB1*246, DPB1*247, DPB1*248, DPB1*249, DPB1*25, DPB1*250, DPB1*251, DPB1*252, DPB1*253, DPB1*254, DPB1*255, DPB1*256, DPB1*257, DPB1*258, DPB1*259, DPB1*26, DPB1*260, DPB1*261, DPB1*262, DPB1*263, DPB1*264, DPB1*265, DPB1*266, DPB1*267, DPB1*268, DPB1*269, DPB1*27, DPB1*270, DPB1*271, DPB1*272, DPB1*273, DPB1*274, DPB1*275, DPB1*276, DPB1*277, DPB1*278, DPB1*279, DPB1*28, DPB1*280, DPB1*281, DPB1*282, DPB1*283, DPB1*284, DPB1*285, DPB1*286, DPB1*287, DPB1*288, DPB1*289, DPB1*29, DPB1*290, DPB1*291, DPB1*292, DPB1*293, DPB1*294, DPB1*295, DPB1*296, DPB1*297, DPB1*298, DPB1*299, DPB1*30, DPB1*301, DPB1*302, DPB1*303, DPB1*304, DPB1*305, DPB1*306, DPB1*307, DPB1*308, DPB1*309, DPB1*31, DPB1*310, DPB1*311, DPB1*312, DPB1*313, DPB1*314, DPB1*315, DPB1*316, DPB1*317, DPB1*318, DPB1*319, DPB1*32, DPB1*320, DPB1*321, DPB1*322, DPB1*323, DPB1*324, DPB1*325, DPB1*326, DPB1*327, DPB1*328, DPB1*329, DPB1*33, DPB1*330, DPB1*331, DPB1*332, DPB1*333, DPB1*334, DPB1*335, DPB1*336, DPB1*337, DPB1*338, DPB1*339, DPB1*34, DPB1*340, DPB1*341, DPB1*342, DPB1*343, DPB1*344, DPB1*345, DPB1*346, DPB1*347, DPB1*348, DPB1*349, DPB1*35, DPB1*350, DPB1*351, DPB1*352, DPB1*353, DPB1*354, DPB1*355, DPB1*356, DPB1*357, DPB1*358, DPB1*359, DPB1*36, DPB1*360, DPB1*361, DPB1*362, DPB1*363, DPB1*364, DPB1*365, DPB1*366, DPB1*367, DPB1*368, DPB1*369, DPB1*37, DPB1*370, DPB1*371, DPB1*372, DPB1*373, DPB1*374, DPB1*375, DPB1*376, DPB1*377, DPB1*378, DPB1*379, DPB1*38, DPB1*380, DPB1*381, DPB1*382, DPB1*383, DPB1*384, DPB1*385, DPB1*386, DPB1*387, DPB1*388, DPB1*389, DPB1*39, DPB1*390, DPB1*391, DPB1*392, DPB1*393, DPB1*394, DPB1*395, DPB1*396, DPB1*397, DPB1*398, DPB1*399, DPB1*40, DPB1*401, DPB1*402, DPB1*403, DPB1*404, DPB1*405, DPB1*406, DPB1*407, DPB1*408, DPB1*409, DPB1*41, DPB1*410, DPB1*411, DPB1*412, DPB1*413, DPB1*414, DPB1*415, DPB1*416, DPB1*417, DPB1*418, DPB1*419, DPB1*42, DPB1*420, DPB1*421, DPB1*422, DPB1*423, DPB1*424, DPB1*425, DPB1*426, DPB1*427, DPB1*428, DPB1*429, DPB1*43, DPB1*430, DPB1*431, DPB1*432, DPB1*433, DPB1*434, DPB1*435, DPB1*436, DPB1*437, DPB1*438, DPB1*439, DPB1*44, DPB1*440, DPB1*441, DPB1*442, DPB1*443, DPB1*444, DPB1*445, DPB1*446, DPB1*447, DPB1*448, DPB1*449, DPB1*45, DPB1*450, DPB1*451, DPB1*452, DPB1*453, DPB1*454, DPB1*455, DPB1*456, DPB1*457, DPB1*458, DPB1*459, DPB1*46, DPB1*460, DPB1*461, DPB1*462, DPB1*463, DPB1*464, DPB1*465, DPB1*466, DPB1*467, DPB1*468, DPB1*469, DPB1*47, DPB1*470, DPB1*471, DPB1*472, DPB1*473, DPB1*474, DPB1*475, DPB1*476, DPB1*477, DPB1*478, DPB1*479, DPB1*48, DPB1*480, DPB1*481, DPB1*482, DPB1*483, DPB1*484, DPB1*485, DPB1*486, DPB1*487, DPB1*488, DPB1*489, DPB1*49, DPB1*490, DPB1*491, DPB1*492, DPB1*493, DPB1*494, DPB1*495, DPB1*496, DPB1*497, DPB1*498, DPB1*499, DPB1*50, DPB1*501, DPB1*502, DPB1*503, DPB1*504, DPB1*505, DPB1*506, DPBDPB1*194, DPB1*195, DPB1*196, DPB1*197, DPB1*198, DPB1*199, DPB1*20, DPB1*200, DPB1*201, DPB1*202, DPB1*203, DPB1*204, DPB1*205, DPB1*206, DPB1*207, DPB1*208, DPB1*209, DPB1*21, DPB1*210, DPB1*211, DPB1*212, DPB1*213, DPB1*214, DPB1*215, DPB1*216, DPB1*217, DPB1*218, DPB1*219, DPB1*22, DPB1*220, DPB1*221, DPB1*222, DPB1*223, DPB1*224, DPB1*225, DPB1*226, DPB1*227, DPB1*228, DPB1*229, DPB1*23, DPB1*230, DPB1*231, DPB1*232, DPB1*233, DPB1*234, DPB1*235, DPB1*236, DPB1*237, DPB1*238, DPB1*239, DPB1*24, DPB1*240, DPB1*241, DPB1*242, DPB1*243, DPB1*244, DPB1*245, DPB1*246, DPB1*247, DPB1*248, DPB1*249, DPB1*25, DPB1*250, DPB1*251, DPB1*252, DPB1*253, DPB1*254, DPB1*255, DPB1*256, DPB1*257, DPB1*258, DPB1*259, DPB1*26, DPB1*260, DPB1*261, DPB1*262, DPB1*263, DPB1*264, DPB1*265, DPB1*266, DPB1*267, DPB1*268, DPB1*269, DPB1*27, DPB1*270, DPB1*271, DPB1*272, DPB1*273, DPB1*274, DPB1*275, DPB1*276, DPB1*277, DPB1*278, DPB1*279, DPB1*28, DPB1*280, DPB1*281, DPB1*282, DPB1*283, DPB1*284, DPB1*285, DPB1*286, DPB1*287, DPB1*288, DPB1*289, DPB1*29, DPB1*290, DPB1*291, DPB1*292, DPB1*293, DPB1*294, DPB1*295,DPB1*296, DPB1*297, DPB1*298, DPB1*299, DPB1*30, DPB1*300, DPB1*301, DPB1*302, DPB1*303, DPB1*304, DPB1*305, DPB1*306, DPB1*307, DPB1*308, DPB1*309, DPB1*31, DPB1*310, DPB1*311, DPB1*312, DPB1*313, DPB1*314, DPB1*315, DPB1*316, DPB1*317, DPB1*318, DPB1*319, DPB1*32, DPB1*320, DPB1*321, DPB1*322, DPB1*323, DPB1*324, DPB1*325, DPB1*326, DPB1*327, DPB1*328, DPB1*329, DPB1*33, DPB1*330, DPB1*331, DPB1*332, DPB1*333, DPB1*334, DPB1*335, DPB1*336, DPB1*337, DPB1*338, DPB1*339, DPB1*34, DPB1*340, DPB1*341, DPB1*342, DPB1*343, DPB1*344, DPB1*345, DPB1*346, DPB1*347, DPB1*348, DPB1*349, DPB1*35, DPB1*350, DPB1*351, DPB1*352, DPB1*353, DPB1*354, DPB1*355, DPB1*356, DPB1*357, DPB1*358, DPB1*359, DPB1*36, DPB1*360, DPB1*361, DPB1*362, DPB1*363, DPB1*364, DPB1*365, DPB1*366, DPB1*367, DPB1*368, DPB1*369, DPB1*37, DPB1*370, DPB1*371, DPB1*372, DPB1*373, DPB1*374, DPB1*375, DPB1*376, DPB1*377, DPB1*378, DPB1*379, DPB1*38, DPB1*380, DPB1*381, DPB1*382, DPB1*383, DPB1*384, DPB1*385, DPB1*386, DPB1*387, DPB1*388, DPB1*389, DPB1*39, DPB1*390, DPB1*391, DPB1*392, DPB1*393, DPB1*394, DPB1*395, DPB1*396, DPB1*397,DPB1*398, DPB1*399, DPB1*40, DPB1*400, DPB1*401, DPB1*402, DPB1*403, DPB1*404, DPB1*405, DPB1*406, DPB1*407, DPB1*408, DPB1*409, DPB1*41, DPB1*410, DPB1*411, DPB1*412, DPB1*413, DPB1*414, DPB1*415, DPB1*416, DPB1*417, DPB1*418, DPB1*419, DPB1*420, DPB1*421, DPB1*422, DPB1*423, DPB1*424, DPB1*425, DPB1*426, DPB1*427, DPB1*428, DPB1*429, DPB1*430, DPB1*431, DPB1*432, DPB1*433, DPB1*434, DPB1*435, DPB1*436, DPB1*437, DPB1*438, DPB1*439, DPB1*44, DPB1*440, DPB1*441, DPB1*442, DPB1*443, DPB1*444, DPB1*445, DPB1*446, DPB1*447, DPB1*448, DPB1*449, DPB1*45, DPB1*450, DPB1*451, DPB1*452, DPB1*453, DPB1*454, DPB1*455, DPB1*456, DPB1*457, DPB1*458, DPB1*459, DPB1*46, DPB1*460, DPB1*461, DPB1*462, DPB1*463, DPB1*464, DPB1*465, DPB1*466, DPB1*467, DPB1*468, DPB1*469, DPB1*47, DPB1*470, DPB1*471, DPB1*472, DPB1*473, DPB1*474, DPB1*475, DPB1*476, DPB1*477, DPB1*478, DPB1*479, DPB1*48, DPB1*480, DPB1*481, DPB1*482, DPB1*483, DPB1*484, DPB1*485, DPB1*486, DPB1*487, DPB1*488, DPB1*489, DPB1*49, DPB1*490, DPB1*491, DPB1*492, DPB1*493, DPB1*494, DPB1*495, DPB1*496, DPB1*497, DPB1*498, DPB1*499, DPB1*50, DPB1*500,DPB1*501, DPB1*502, DPB1*503, DPB1*504, DPB1*505, DPB1*506, DPB1*507, DPB1*508, DPB1*509, DPB1*51, DPB1*510, DPB1*511, DPB1*512, DPB1*513, DPB1*514, DPB1*515, DPB1*516, DPB1*517, DPB1*518, DPB1*519, DPB1*52, DPB1*520, DPB1*521, DPB1*522, DPB1*523, DPB1*524, DPB1*525, DPB1*526, DPB1*527, DPB1*528, DPB1*529, DPB1*53, DPB1*530, DPB1*531, DPB1*532, DPB1*533, DPB1*534, DPB1*535, DPB1*536, DPB1*537, DPB1*538, DPB1*539, DPB1*54, DPB1*540, DPB1*541, DPB1*542, DPB1*543, DPB1*544, DPB1*545, DPB1*546, DPB1*547, DPB1*548, DPB1*549, DPB1*55, DPB1*550, DPB1*551, DPB1*552, DPB1*553, DPB1*554, DPB1*555, DPB1*556, DPB1*557, DPB1*558, DPB1*559, DPB1*56, DPB1*560, DPB1*561, DPB1*562, DPB1*563, DPB1*564, DPB1*565, DPB1*566, DPB1*567, DPB1*568, DPB1*569, DPB1*57, DPB1*570, DPB1*571, DPB1*572, DPB1*573, DPB1*574, DPB1*575, DPB1*576, DPB1*577, DPB1*578, DPB1*579, DPB1*58, DPB1*580, DPB1*581, DPB1*582, DPB1*583, DPB1*584, DPB1*585, DPB1*586, DPB1*587, DPB1*588, DPB1*589, DPB1*59, DPB1*590, DPB1*591, DPB1*592, DPB1*593, DPB1*594, DPB1*595, DPB1*596, DPB1*597, DPB1*598, DPB1*599, DPB1*60, DPB1*600, DPB1*601, DPB1*602,DPB1*603, DPB1*604, DPB1*605, DPB1*606, DPB1*607, DPB1*608, DPB1*609, DPB1*61, DPB1*610, DPB1*611, DPB1*612, DPB1*613, DPB1*614, DPB1*615, DPB1*616, DPB1*617, DPB1*618, DPB1*619, DPB1*62, DPB1*620, DPB1*621, DPB1*622, DPB1*623, DPB1*624, DPB1*625, DPB1*626, DPB1*627, DPB1*628, DPB1*629, DPB1*63, DPB1*630, DPB1*631, DPB1*632, DPB1*633, DPB1*634, DPB1*635, DPB1*636, DPB1*637, DPB1*638, DPB1*639, DPB1*64, DPB1*640, DPB1*641, DPB1*642, DPB1*643, DPB1*644, DPB1*645, DPB1*646, DPB1*647, DPB1*648, DPB1*649, DPB1*65, DPB1*650, DPB1*651, DPB1*652, DPB1*653, DPB1*654, DPB1*655, DPB1*656, DPB1*657, DPB1*658, DPB1*659, DPB1*66, DPB1*660, DPB1*661, DPB1*662, DPB1*663, DPB1*664, DPB1*665, DPB1*666, DPB1*667, DPB1*668, DPB1*669, DPB1*67, DPB1*670, DPB1*671, DPB1*672, DPB1*673, DPB1*674, DPB1*675, DPB1*676, DPB1*677, DPB1*678, DPB1*679, DPB1*68, DPB1*680, DPB1*681, DPB1*682, DPB1*683, DPB1*684, DPB1*685, DPB1*686, DPB1*687, DPB1*688, DPB1*689, DPB1*69, DPB1*690, DPB1*691, DPB1*692, DPB1*693, DPB1*694, DPB1*695, DPB1*696, DPB1*697, DPB1*698, DPB1*699, DPB1*70, DPB1*700, DPB1*701, DPB1*702, DPB1*703, DPB1*704,DPB1*705, DPB1*706, DPB1*707, DPB1*708, DPB1*709, DPB1*71, DPB1*710, DPB1*711, DPB1*712, DPB1*713, DPB1*714, DPB1*715, DPB1*716, DPB1*717, DPB1*718, DPB1*719, DPB1*72, DPB1*720, DPB1*721, DPB1*722, DPB1*723, DPB1*724, DPB1*725, DPB1*726, DPB1*727, DPB1*728, DPB1*729, DPB1*73, DPB1*730, DPB1*731, DPB1*732, DPB1*733, DPB1*734, DPB1*735, DPB1*736, DPB1*737, DPB1*738, DPB1*739, DPB1*74, DPB1*740, DPB1*741, DPB1*742, DPB1*743, DPB1*744, DPB1*745, DPB1*746, DPB1*747, DPB1*748, DPB1*749, DPB1*75, DPB1*750, DPB1*751, DPB1*752, DPB1*753, DPB1*754, DPB1*755, DPB1*756, DPB1*757, DPB1*758, DPB1*759, DPB1*76, DPB1*760, DPB1*761, DPB1*762, DPB1*763, DPB1*764, DPB1*765, DPB1*766, DPB1*767, DPB1*768, DPB1*769, DPB1*77, DPB1*770, DPB1*771, DPB1*772, DPB1*773, DPB1*774, DPB1*775, DPB1*776, DPB1*777, DPB1*778, DPB1*779, DPB1*78, DPB1*780, DPB1*781, DPB1*782, DPB1*783, DPB1*784, DPB1*785, DPB1*786, DPB1*787, DPB1*788, DPB1*789, DPB1*79, DPB1*790, DPB1*791, DPB1*792, DPB1*794, DPB1*795, DPB1*796, DPB1*797, DPB1*798, DPB1*799, DPB1*80, DPB1*800, DPB1*801, DPB1*802, DPB1*803, DPB1*804, DPB1*805, DPB1*806, DPB1*807,DPB1*808, DPB1*809, DPB1*81, DPB1*810, DPB1*811, DPB1*812, DPB1*813, DPB1*814, DPB1*815, DPB1*816, DPB1*817, DPB1*818, DPB1*819, DPB1*82, DPB1*820, DPB1*821, DPB1*822, DPB1*823, DPB1*824, DPB1*825, DPB1*826, DPB1*827, DPB1*828, DPB1*829, DPB1*83, DPB1*830, DPB1*831, DPB1*832, DPB1*833, DPB1*834, DPB1*835, DPB1*836, DPB1*837, DPB1*838, DPB1*839, DPB1*84, DPB1*840, DPB1*841, DPB1*842, DPB1*843, DPB1*844, DPB1*845, DPB1*846, DPB1*847, DPB1*848, DPB1*849, DPB1*85, DPB1*850, DPB1*851, DPB1*852, DPB1*853, DPB1*854, DPB1*855, DPB1*856, DPB1*857, DPB1*858, DPB1*859, DPB1*86, DPB1*860, DPB1*861, DPB1*862, DPB1*863, DPB1*864, DPB1*865, DPB1*866, DPB1*867, DPB1*868, DPB1*869, DPB1*87, DPB1*870, DPB1*871, DPB1*872, DPB1*873, DPB1*874, DPB1*875, DPB1*876, DPB1*877, DPB1*878, DPB1*879, DPB1*88, DPB1*880, DPB1*881, DPB1*882, DPB1*883, DPB1*884, DPB1*885, DPB1*886, DPB1*887, DPB1*888, DPB1*889, DPB1*89, DPB1*890, DPB1*891, DPB1*892, DPB1*893, DPB1*894, DPB1*895, DPB1*896, DPB1*897, DPB1*898, DPB1*899, DPB1*90, DPB1*900, DPB1*901, DPB1*902, DPB1*903, DPB1*904, DPB1*905, DPB1*906, DPB1*907, DPB1*908, DPB1*909,DPB1*91, DPB1*910, DPB1*911, DPB1*912, DPB1*913, DPB1*914, DPB1*915, DPB1*916, DPB1*917, DPB1*918, DPB1*919, DPB1*92, DPB1*920, DPB1*921, DPB1*922, DPB1*923, DPB1*924, DPB1*925, DPB1*926, DPB1*927, DPB1*928, DPB1*929, DPB1*93, DPB1*930, DPB1*931, DPB1*932, DPB1*933, DPB1*934, DPB1*935, DPB1*936, DPB1*937, DPB1*938, DPB1*939, DPB1*94, DPB1*940, DPB1*941, DPB1*942, DPB1*943, DPB1*944, DPB1*945, DPB1*946, DPB1*947, DPB1*948, DPB1*949, DPB1*95, DPB1*950, DPB1*951, DPB1*952, DPB1*953, DPB1*954, DPB1*955, DPB1*956, DPB1*957, DPB1*958, DPB1*959, DPB1*96, DPB1*960, DPB1*961, DPB1*962, DPB1*963, DPB1*964, DPB1*965, DPB1*97, DPB1*98, and DPB1*99.

[0024] In some aspects, the DPB chain comprises the amino acid sequence set forth in SEQ ID NO: 1.

[0025] In some aspects, the HLA class II molecule further comprises a DP a chain. In some aspects, the DP a chain is selected from DPA1*01:03:01:01, DPA1*01:03:01:02, DPA1*01:03:01:03, DPA1*01:03:01:04, DPA1*01:03:01:05, DPA1*01:03:01:06, DPA1*01:03:01:07, DPA1*01:03:01:08, DPA1*01:03:01:09, DPA1*01:03:01:10, DPA1*01:03:01:11, DPA1*01:03:01:12, DPA1*01:03:01:13, DPA1*01:03:01:14, DPA1*01:03:01:15, DPA1*01:03:01:16, DPA1*01:03:01:17, DPA1*01:03:01:18Q, DPA1*01:03:01:19, DPA1*01:03:01:20, DPA1*01:03:01:21, DPA1*01:03:01:22, DPA1*01:03:01:23, DPA1*01:03:02, DPA1*01:03:03, DPA1*01:03:04, DPA1*01:03:05, DPA1*01:03:06, DPA1*01:03:07, DPA1*01:03:08, DPA1*01:03:09, DPA1*01:04, DPA1*01:05, DPA1*01:06:01, DPA1*01:06:02, DPA1*01:07, DPA1*01:08, DPA1*01:09, DPA1*01:10, DPA1*01:11, DPA1*01:12, DPA1*01:13, DPA1*01:14, DPA1*01:15, DPA1*01:16, DPA1*01:17, DPA1*01:18, DPA1*01:19, DPA1*02:01:01:01, DPA1*02:01:01:02, DPA1*02:01:01:03, DPA1*02:01:01:04, DPA1*02:01:01:05, DPA1*02:01:01:06, DPA1*02:01:01:07, DPA1*02:01:01:08, DPA1*02:01:01:09, DPA1*02:01:01:10, DPA1*02:01:01:11, DPA1*02:01:02:01, DPA1*02:01:02:02, DPA1*02:01:03, DPA1*02:01:04, DPA1*02:01:05,DPA1*02:01:06, DPA1*02:01:07, DPA1*02:01:08:01, DPA1*02:01:08:02, DPA1*02:02:02:01, DPA1*02:02:02:02, DPA1*02:02:02:03, DPA1*02:02:02:04, DPA1*02:02:02:05, DPA1*02:02:03, DPA1*02:02:04, DPA1*02:02:05, DPA1*02:02:06, DPA1*02:03, DPA1*02:04, DPA1*02:05, DPA1*02:06, DPA1*02:07:01:01, DPA1*02:07:01:02, DPA1*02:07:01:03, DPA1*02:08, DPA1*02:09, DPA1*02:10, DPA1*02:11, DPA1*02:12, DPA1*02:13N, DPA1*02:14, DPA1*02:15, DPA1*02:16, DPA1*03:01:01:01, DPA1*03:01:01:02, DPA1*03:01:01:03, DPA1*03:01:01:04, DPA1*03:01:01:05, DPA1*03:01:02, DPA1*03:02, DPA1*03:03, DPA1*03:04, DPA1*04:01:01:01, DPA1*04:01:01:02, and DPA1*04:01:01:03, DPA1*04:02.

[0026] In some aspects, the DPa 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 to SEQ ID NO: 6 or 8. In some aspects, the DPa chain comprises the amino acid sequence set forth in SEQ ID NO: 6 or 8.

[0027] In some aspects, the HLA Class II molecule is a DP1, DP2, DP3, DP4, DP5, DP6, DP8, or DP9 allele.

[0028] In some aspects, the DPp chain has increased affinity for a CD4 protein compared to a reference HLA Class II molecule, wherein the DPp chain comprised by the reference HLA Class II molecule comprises (i) a leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 and / or (ii) a valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1.

[0029] In some aspects, the increased affinity is at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, at least about 50-fold, at least about 75-fold, at least about 100-fold, at least about 200-fold, at least about 300-fold, at least about 400-fold, at least about 500-fold, or at least about 1000-fold.

[0030] In some aspects, the DPβ chain is associated with the membrane of a cell. In some aspects, the DPβ chain is not associated with the membrane of a cell. In some aspects, the DPβ chain comprises the extracellular domain of a full-length DPα chain. In some aspects, the DPβ chain does not comprise the transmembrane domain of a full-length DPβ chain.

[0031] In some aspects, the DPα chain is associated with the membrane of a cell. In some aspects, the DPα chain is not associated with the membrane of a cell. In some aspects, the DPα chain comprises the extracellular domain of a full-length DPα chain. In some aspects, the DPα chain does not comprise the transmembrane domain of a full-length DPα chain.

[0032] In some aspects, the DPβ chain is attached to or associated with an inert particle. In some aspects, the inert particle is a bead. In some aspects, the inert particle is a nanoparticle. In some aspects, the nanoparticle is selected from the group consisting of pegylated iron oxide, chitosan, dextran, gelatin, alginate, liposome, starch, branched polymer, carbon-based carrier, polylactic acid, poly(cyano)acrylate, polyethylenimine, block copolymer, polycaprolactone, SPIONS, USPIONS, Cd / Zn-selenide, or silica nanoparticle. In some aspects, the nanoparticle is a pegylated iron oxide nanoparticle.

[0033] In some aspects, the DPβ chain comprises a signal peptide. In some aspects, the DPα chain comprises a signal peptide. In some aspects, the signal peptide comprises the amino acid sequence set forth in SEQ ID NO: 9.

[0034] Certain aspects of the present disclosure relate to a nucleic acid molecule encoding a DPβ chain disclosed herein. In some aspects, the nucleic acid molecule further encodes a DPα chain disclosed herein.

[0035] In some aspects, 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 to SEQ ID NO: 2.

[0036] Certain aspects of the present disclosure relate to a vector comprising a nucleic acid molecule disclosed herein.

[0037] Certain aspects of the present disclosure relate to a cell comprising an HLA class II molecule disclosed herein, a nucleic acid molecule disclosed herein, or a vector disclosed herein. In some aspects, the cell is a mammalian cell or an insect cell. In some aspects, the cell is selected from a K562 cell, T2, HEK293, HEK293T, A375, SK-MEL-28, Me275, COS, fibroblast, tumor cell, or any combination thereof.

[0038] In some aspects, the cell lacks endogenous MHC class II DP beta chain expression. In some aspects, the cell lacks endogenous MHC class II DP alpha chain expression.

[0039] Certain aspects of the present disclosure relate to a method of identifying a T cell receptor capable of binding an epitope in an MHC class II complex, the method comprising pulsing a cell disclosed herein with one or more peptides comprising the epitope, and stimulating one or more CD4 + T cells.

[0040] Certain aspects of the present disclosure relate to a method of treating a disease or condition in a subject in need thereof, the method comprising administering to the subject an MHC class II molecule disclosed herein. In some aspects, the disease or condition is a cancer or an infection.

[0041] In some aspects, the 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 ureter cancer, renal pelvis cancer, glioma, squamous cell carcinoma, and a combination of said cancers.

[0042] In some aspects, the cancer is relapsed or refractory. In some aspects, the cancer is locally advanced. In some aspects, the cancer is advanced. In some aspects, the cancer is metastatic.

[0043] In some aspects, the HLA class II molecule binds CD4 with a K D In some aspects, the HLA class II molecule binds CD4 with a K D In some aspects, the HLA class II molecule binds CD4 with a K D In some aspects, the HLA class II molecule binds CD4 with a K

[0044] Certain aspects of the present disclosure relate to complexes comprising an HLA class II molecule disclosed herein and a peptide, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 32-237. BRIEF DESCRIPTION OF DRAWINGS

[0045] Figures 1A-1V is a histogram showing the results of affinity maturation of DP4 L112W / V141M molecules exhibiting enhanced CD4 binding ability. Figures 1A-1F is a histogram showing the results of stable HLA class II-null K562 cells expressing wild-type DP alpha chain (DPAl*01:03) transduced with empty, wild-type or mutant DP beta chains containing L112W, V114M, V141M and M158I substitutions (DP4 L112W / V114M / V141M / M158I ) and stained with anti-class II mAb and soluble CD4 (sCD4). Figure 1G is a bar graph summarizing the binding affinity (MFI; y-axis) of all possible DP4 back-mutants to sCD4, which were similarly expressed and stained with sCD4. As quantified by steady-state analysis, Figures 1A-1F Figure 1H shows the affinity between DP4 L112W / V141M and CD4. Figure 1I shows the results of IL-2 EPISPOT assays of Jurkat 76 and Jurkat 76 / CD4 cells transduced with DP4 / WT1 TCR clone 9 stimulated by aAPCs expressing wild-type DP4 or DP4 L112W / V141M . Figures 1J-1W is a histogram showing K562 cells expressing DP L112W / V141M alleles (as indicated) stained with anti-class II mAb and sCD4. Open histograms represent isotype control staining. *P < 0.05 by Student’s t-test. Bars and error bars represent the mean ± SD of results from experiments performed in triplicate. At least 2 independent experiments were performed. Figures 1X-1AA is a histogram showing wild-type DP4 and DP4 L112W / V141M molecules on the surface of K562 cells detected with the indicated anti-HLA class II antibodies. Staining of control cells without class II expression is shown as a solid gray color. Figure 1A B to Figure 1B H is a histogram showing indicated concentrations of aAPCs expressing indicated DP4 or class II parental cells stained with sCD4. Figure 1BI shows quantification of indicated concentrations of aAPCs expressing wild-type DP4 or DP4 L112W / V141M . Error bars represent the mean ± standard deviation of experiments performed in triplicate.​Figure 1BJ is a biolayer interferometry sensorgram showing the interaction of biotinylated wild-type DP4 (ligand) with sCD4 (analyte) over a range of concentrations. Figure 1BK is a biolayer interferometry sensorgram showing the interaction of biotinylated DP4 L112W / V141M (ligand) with sCD4 (analyte) over a range of concentrations. Figure 1BJ and 1BI were performed in parallel. All data represent two independent experiments.

[0046] Figures 2A-2D is a ribbon diagram of the model structure of DP4 L112W / V141M and human CD4 complex. Figures 2A-2B is a two-directional view of the model structure of the ternary complex of DPA1*01:03, DPB1*04:01 and CD4, as indicated. The DPB1*04:01-CD4 binding interface is boxed with a dashed line ( Figure 2B ). Figures 2C-2D Close-up views of the CD4 binding interface of wild-type DP4 ( Figure 2C ) and DP4 L112W / V141M ( Figure 2D ) are provided. Side chains of interacting residues are shown as stick representations ( Figures 2C-2D ).

[0047] Figures 3A-3P are graphical representations of data illustrating DP4 L112W / V141M dimer staining of cognate TCR expressed in human primary CD4 + T cells. Primary T cells were transduced with DP4 / MAGE-A3 243-258 (R12C9; Figures 3E-3H), DP4 / WT1 328-348 (clonal 9; Figures 31-3L) or DP4 / NY-ESO-1 157-170 (5B8; Figures 3M-3P) TCR and stained with the indicated DP4 L112W / V141M dimer (Figures 3B-3D, 3F-3H, 3J-3L and 3N-3P).

[0048] Figures 4A-4D are scatter plots illustrating the composite staining of R12C9 transduced CD4 L112W / V141M T cells stained with DP4 + dimer and anti-Vp22 mAb. Note that R12C9 represents Vp22. Figures 4E-4H are scatter plots illustrating the composite staining of clonal 9 transduced CD4 L112W / V141M T cells double stained with DP4 + dimer and anti-NGFR mAb. Note that clonal 9 and the NGFR gene are fused with P2A.

[0049] Figures 5A to 5P illustrate the use of 5 μg / ml conventional wild-type DP4 tetramer and DP4 L112W / V141M Scatter plots of primary T cells stained with a combination of clone 9 transduction (Fig. 5A to 5H) and 5B8 transduction (Fig. 5I to 5P) for dimer staining. Perform at least two independent experiments.

[0050] Figures 6A-6F This explains the use of DP4 L112W / V141M A bar graph showing the results of comprehensive screening of dimers, which identified a range of novel DP4-restricted tumor-associated antigens. From 6 DP4... + Purified peripheral CD4 from melanoma patients + T cells were stimulated with aAPCs expressing DP4 by shocks with 196 different peptides derived from tumor-associated antigens, and homologous DP4 was used. L112W / V141M Dimer staining. Results were displayed using the 30 peptides with the highest positive values. Figures 6A-6B The results for the remaining 166 peptides were shown in... Figures 6C-6F In the middle. Set each gate so that the control dimer staining shows <0.2% positivity. Positive dimer staining is defined as staining exceeding 3 standard deviations of control dimer staining, as shown by the dashed line (>0.6%).

[0051] Figures 7A-7L It is a peptide-specific CD4 from melanoma patients. + DP4 of T cells L112W / V141M Graphical representation of dimer staining. From 6 DP4 + Purified primary CD4 from melanoma patients + T cells were stimulated with aAPCs expressing DP4 by shocks with 196 different peptides derived from tumor-associated antigens, and homologous DP4 was used. L112W / V141M Dimer staining, such as Figures 6A-6F As shown. DP4 is displayed. L112W / V141M Example of dimer staining. *P<0.05, obtained by Student's t-test. ns, not significant. Perform at least two independent experiments.

[0052] Figures 8A to 8X are graphical representations of the data, illustrating the changes from DP4. L112W / V141M Isolate from dimeric positive cells and in human TCR-deficient CD4 + The reconstructed DP4-restricted TCR in T cells functions in a DP4-restricted and antigen-specific manner. 03-CCND1 219-238 (Figures 8A to 8D), 05-HSD17B12 225-244 and 09-HSD17B12 225-244(FIG. 8E to FIG. 8J), 05-LGSN 296-315 (FIG. 8K to FIG. 8N), 03-MAGE-A2 108-127 and 06-MAGE-A2 108-127 (FIG. 8O to FIG. 8T) and 05-MUC5AC 4922-4941 (FIG. 8U to FIG. 8X) from DP4 L112W / V141M dimer-positive cell clones were reconstituted in TCR-deficient Jurkat 76 / CD4 cells and stained with the corresponding DP4 L112W / V141M dimers.

[0053] Figures 9A-9G are bar graphs illustrating the results of IL-2 ELISPOT assays of 03-CCND1 219-238 ( Figure 9A ), 05-HSD17B12 225-244 ( Figure 9B ), 09-HSD17B12 225-244 ( Figure 9C ), 05-LGSN 296-315 ( Figure 9D ), 03-MAGE-A2 108-127 ( Figure 9E ), 06-MAGE-A2 108-127 ( Figure 9F ) and 05-MUC5AC 4922-4941 ( Figure 9G ) stimulated by aAPCs pulsed with the corresponding peptides. DP4 / WT1 (clone 9) TCR was used as a negative control. At least two independent experiments were performed. *, P < 0.05 by Student’s t-test. Bars and error bars represent the mean ± SD of triplicate experiments.

[0054] FIG. 10A to Figure 10Q are graphical representations of data showing that DP4-restricted TCRs isolated from DP4 L112W / V141M dimer-positive cells and reconstituted in human primary CD4 + T cells act in a DP4-restricted and antigen-specific manner. 03-CCND1 219-238 (FIG. 10A to FIG. 10D and Figure 10O ), 03-MAGE-A2 108-127 and 06-MAGE-A2 108-127 (FIG. 10E to FIG. 10J and Figure 10P ) and 05-MUC5AC 4922-4941 (FIG. 10K to FIG. 10N and Figure 10Q ) were retrovirally transduced into human primary CD4 + T cells and stained with the corresponding DP4 L112W / V141MDimer staining (Figs. 10A to 10N). *P<0.05, obtained by Student's t-test. ns, not significant. Perform at least two independent experiments. *,P<0.05, obtained by Student's t-test. Bars and error bars represent the mean ± SD of three experimental results.

[0055] Figure 11A The data presented in Figure 11E show that DP4-restricted TCRs from melanoma patient clones recognize peptides that are endogenously processed and presented by K562-based aAPCs. Figures 11A-11B This demonstrates the endogenous expression of CCDN1 in K562-derived aAPC cells. Figure 11A ) and MAGE-A2 ( Figure 11B Images of gel chromatography. Figures 11C-11D It shows 03-CCND1 219-238 ( Figure 11C ) or 06-MAGE-A2 108-127 ( Figure 11D Bar graph showing the results of IFN-γ ELISPOT assays in human primary T cells transduced with retroviral agents and stimulated with HLA-empty or DP4-aAPC cells without peptide shock. Figures 11C-11D Figure 11E shows the use of 05-MUC5AC. 4922-4941 TCR retroviral transduction and MUC5AC 4914-4949 Bar graphs of IFN-γ ELISPOT assays in small gene transduction and HLA-empty or DP4-aAPC-unstimulated human primary T cells without peptide shock. At least two independent experiments were performed. *, P < 0.05, obtained via Student's t-test. Bars and error bars represent the mean ± SD of three experimental results.

[0056] Figures 12A-12E The presented data is 06-MAGE-A2 108-127 TCR recognizes melanoma cell lines in a DP4 and MAGE-A2 dependent manner. Figure 12A This is a Western blot image showing endogenous MAGE-A2 expression in K562 cells and the melanoma cell line shown. Figures 12B-12E It's a bar chart showing results from using SK-MEL-21 (DP4) + MAGE-A2 - ; Figure 12B ) or SK-MEL-37 (DP4) + MAGE-A2 + ; Figure 12C Stimulating with 06-MAGE-A2 108-127 TCR-transduced primary human T cells and those using SK-MEL-28 (DP4) -MAGE-A2 + ; Figure 12D ) and Me275 (DP4 - MAGE-A2 + ; Figure 12E ) stimulated IFN-γ ELISPOT assays with DP4 transduced cases. *, P < 0.05 by Student's t test. Bars and error bars represent mean ± SD of triplicate experimental results. At least 2 independent experiments were performed.

[0057] Figures 13A-13Q is a histogram comparing expression levels of wild-type HLA DP*04:01 and its derivatives in K562 cells stained with anti-HLA class II mAb clone 9-49. Open histograms represent isotype control staining. Figures 14A-14D is a graphical representation showing comparison of DP4 L112W / V141M dimer and dextramer to endogenous TRPC1 578-597 specific CD4 + T cells. CD4 + T cells from melanoma patients were expanded with endogenous (non-transduced) TRPC1 578-597 specific CD4 + T cells stimulated with DP4 L112W / V141M TRPC1 578-597 dimer ( Figure 14B ) or TRPC1 578-597 dextramer (14D). Corresponding CLIP multimers were used as controls ( Figure 14A and Figure 14C ).

[0058] Figures 15A-15F is a graphical representation showing comparison of DP4 L112W / V141M dimer to conventional DP4 tetramer and dextramer to endogenous NY-ESO-1 157-170 specific T cells. CD4 + T cells were purified from DP4 + healthy donor #4 and stimulated once with NY-ESO-1 157-170 pulsed and irradiated DP4 + artificial APC. As shown by three different DP4 multimers (DP4 L112W / V141M dimer ( Figure 15B ), DP4 tetramer (15D) or DP4 dextramer (FIG. 15F), expanded CD4 + T cells were stained, respectively.

[0059] Figures 16A to 16Y show the process of using DP4. L112W / V141M Pathogen-specific CD4 staining in vitro using dimers + Graphical representation of T cell data. From five DP4 + Purified memory CD4 from donor + T cells, targeting the following pathogen-associated peptides with DP4 in the absence of in vitro stimulation. L112W / V141M In vitro staining of dimers: TT 948-968 (Figures 16F to 16J), HSV-2-UL21 283-302 (Figures 16K to 16O), Flu-HA 527-546 (Figures 16P to 16T) and RSV-GP 162-175 (Figures 16U to 16Y). CLIP peptide was used as a negative control (Figures 16A to 16E).

[0060] Figures 17A-17W It is displayed from DP4 L112W / V141M Dimer + Endogenous RSV-GP successfully established in cells 162-175 Specific CD4 + Graphical representation of T cell clone data. From DP4 No. 6. + Purified memory CD4 from donor + T cells, and DP4 without external stimulation. L112W / V141M RSV-GP 162-175 The dimer was stained in vitro. Then, the dimer was cloned by limiting dilution. + CD4 + T cells. Figures 17A-17V This is a graphical representation of representative dimeric staining data from 10 dimeric-positive and 1 dimeric-negative single-cell clones. 77 of the 84 clones (91.7%) were successfully stained with DP4. L112W / V141M RSV-GP 162-175 Dimer staining. Figure 17W It displays RSV-GP 162-175 Dimer + Bar graph showing antigen-specific IL-2 production in a single-cell clone.

[0061] Figures 18A-18S It is displayed from DP4 L112W / V141M Dimer + Endogenous DP4 TT successfully established in cells 948-968 Specific CD4 + Graphical representation of T cell clone data. From DP4, number 04. + Purified memory CD4 from donor + T cells, and DP4 without external stimulation.L112W / V141M TT 948-968 Dimerization was performed ex vivo. Dimerization was then followed by limiting dilution cloning of dimers + CD4 + T cells. Figures 18A-18R is a graphical representation of representative dimer staining data for 8 dimer positive and 1 dimer negative single cell clones. 26 of 29 clones (89.7%) successfully stained with DP4 L112W / V141M TT 948-968 Dimerization. Figure 18S is a graphical representation of TT 948-968 Dimerization + Bar graph of antigen-specific IL-2 production in single cell clones.

[0062] FIGS. 19A-19NN are graphical representations of DP4 multimer staining of RSV-GP (FIGS. 19A-19P) and TT (FIGS. 19Q-19NN) dimerization + single cell clones. RSV-GP dimers + single cell clones (c6, c12, c26, and c39) were stained with DP4 L112W / V141M RSV-GP 162-175 dimers (FIGS. 19B, 19D, 19F, and 19H) or wild-type DP4 dextramer (FIGS. 19J, 19L, 19N, and 19P). TT dimers + single cell clones (c2, c4, c6, and c9) were stained with three different DP4 TT 948-968 multimers (DP4 L112W / V141M dimers (FIGS. 19R, 19T, 19V, and 19X), wild-type DP4 tetramers (FIGS. 19Z, 19BB, 19DD, and 19FF), and wild-type DP4 dextramer (FIGS. 19HH, 19JJ, 19LL, and 19NN). DETAILED DESCRIPTION

[0063] The present disclosure relates to MHC class II molecules with increased affinity for CD4. In some aspects, the present disclosure relates to MHC class II molecules comprising an HLA-DP (DP) beta chain, wherein the DP beta chain has increased affinity for CD4.

[0064] The present disclosure further relates to MHC class II molecules comprising a DP beta chain, wherein the DP beta chain comprises an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1. In some aspects, the DP beta chain further comprises an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1.

[0065] The present disclosure further relates to MHC class II molecules comprising a DP beta chain, wherein the DP beta chain comprises an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1. In some aspects, the DP beta chain further comprises an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1.

[0066] I. Terminology

[0067] To enable a better understanding of the present disclosure, certain terms are first defined. As used throughout this application, each of the following terms shall have the meaning set forth below, unless the context clearly indicates otherwise. Additional definitions are set forth throughout the application.

[0068] It should be noted that the term "a" or "an" entity refers to one or more than one of that entity; for example, "a nucleotide sequence," is understood to mean one or more nucleotide sequences. As such, the terms "a" (or "an"), "one or more," and "at least one" can be used interchangeably herein.

[0069] Furthermore, "and / or" where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term "and / or" as used in a phrase such as "A and / or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and / or" as used in a phrase such as "A, B, and / or C" is intended to encompass each of the following aspects: 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).

[0070] The term "about" is used herein to mean approximately, roughly, around, or in the vicinity of, when used in relation to a numerical value. When the term "about" is used in conjunction with a numerical range, it modifies the range by extending the boundaries above and below the numerical values. Generally, the term "about" is used herein to modify a numerical value by variations of up to and down from the value by 10%.

[0071] It should be appreciated that whenever aspects are described herein with the wording "comprising" various aspects, additional analogous aspects described with the wording "consisting of and / or "consisting essentially of are also provided.

[0072] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure is related. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; the Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary Of Biochemistry And Molecular Biology, Revised Edition, 2000, Oxford University Press, give one of ordinary skill in the art access to a glossary of terms used in the art. Each document, patent, patent application, and / or patent publication cited herein is hereby incorporated by reference in its entirety.

[0073] Units, prefixes, and symbols are denoted in their SI accepted form. Numeric ranges are inclusive of the numbers defining the range. Unless otherwise indicated, nucleotide sequences are written left to right in 5' to 3' orientation. Amino acid sequences are written left to right in amino to carboxyl orientation. The headings provided herein are not limitations of the various aspects of the disclosure, which can be had by reference to the specification as a whole. Accordingly, the terms defined immediately below are more fully defined by reference to the specification as a whole.

[0074] “Administering” refers to the physical introduction of an agent into a subject using any of a variety of methods and delivery systems known to those of skill in the art. Exemplary routes of administration for the formulations disclosed herein include intravenous, intramuscular, subcutaneous, intraperitoneal, spinal or other parenteral routes of administration, e.g., by injection or infusion. As used herein, the phrase“parenteral administration” means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcutaneous, subcutaneous, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion, as well as in vivo electroporation. In some aspects, the formulation is administered by a non-parenteral route, e.g., orally. Other non-parenteral routes include topical, epidermal, or mucosal routes of administration, e.g., intranasal, vaginal, rectal, sublingual, or topical. Administration can also be carried out, for example, once, multiple times, and / or over one or more extended periods of time.

[0075] As used herein, the term“HLA” refers to human leukocyte antigen. In humans, HLA genes encode major histocompatibility complex (MHC) proteins. MHC proteins are expressed on the surface of cells and are involved in activation of immune responses. HLA class II genes encode MHC class II proteins that are expressed on the surface of professional antigen presenting cells (APCs). Non-limiting examples of professional APCs include monocytes, macrophages, dendritic cells (DCs), and B lymphocytes. Some endothelial cells and epithelial cells also express MHC class II molecules upon activation of inflammatory signals. Humans lacking functional MHC class II molecules are highly susceptible to a range of infectious diseases and typically die in early childhood.

[0076] 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 that encodes a MHC class II molecule. Thus,“HLA class II molecule” and“MHC class II molecule” are used interchangeably herein. A typical MHC class II molecule comprises two protein chains: an alpha chain and a beta chain. Generally, each of the naturally occurring alpha and beta chains comprises a transmembrane domain that anchors the alpha / beta chain to the surface of a cell, and an extracellular domain that carries an antigen and interacts with TCRs and / or CD4 expressed on cells.

[0077] Both MHC class II alpha and beta chains are encoded by the HLA gene complex. The HLA complex is located within the 6p21.3 region of human chromosome 6 and comprises more than 220 genes with different functions. It is known in the art that the HLA gene complex is highly variable, with over 20,000 HLA alleles and related alleles, including over 250 MHC class II alpha chain alleles and 5,000 MHC class II beta 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 populations. Each alpha chain and beta chain is typically expressed as a proprotein, which further comprises a signal peptide that is cleaved off. Any number of naturally occurring signal peptides can be used to facilitate expression and localization of the alpha and beta chains disclosed herein. One such example is SEQ ID NO: 9.

[0078] 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-DP sequences are provided in Table 1.

[0079] Table 1: DP beta chain and alpha chain amino acid and nucleotide sequences.

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086] When MHC class II molecules are complexed with an antigenic peptide, an antigenic peptide of 10-30 amino acids in length binds the peptide binding groove and is presented extracellularly to CD4+ cells. Both the alpha and beta chains fold into two separate domains; alpha-1 and alpha-2 of the alpha polypeptide, and beta-1 and beta-2 of the beta polypeptide. The invariant residues at LI 12, VI 14, VI 41, LI 56, and M158 that are recognized and bound by CD4 are located in the beta-2 domain of the beta polypeptide. An open peptide binding groove that holds the presented antigen is found between the alpha-1 and beta-1 domains. In interaction with CD4+ T cells, MHC class II complexes interact with T cell receptors (TCRs) expressed on the surface of T cells. In addition, the beta chain of MHC class II molecules weakly interacts with CD4 expressed on the surface of T cells (K D >2mM). The canonical CD4 amino acid sequence (UniProt-P01730) is provided in Table 2 (SEQ ID NO: 10).

[0087] Table 2: Human CD4 amino acid sequence

[0088]

[0089] As used herein, the term “T cell receptor” (TCR) refers to a heteromeric cell surface receptor capable of specific interaction with a target antigen. As used herein, “TCR” includes, but is not limited to, naturally occurring and non-naturally occurring TCRs, full-length TCRs and antigen-binding portions thereof, 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 of antigen-presenting cells. Antigen-presenting cells (APCs) display fragments of foreign proteins (antigens) complexed with major histocompatibility complexes (MHC class I or MHC class II) (also referred to herein as complexed with HLA molecules, e.g., 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 the T cell, thereby activating the TCR. The activated TCR initiates downstream signaling and immune responses, including destruction of the APC.

[0090] Generally, a TCR can comprise two chains, an alpha chain and a beta chain (or, less commonly, a gamma chain and a delta chain), which are interconnected to each other by disulfide bonds. Each chain comprises a variable region (alpha chain variable domain and beta chain variable domain) and a constant region (alpha chain constant region and beta chain constant region). The variable domain is located distal to the cell membrane, and the variable domain interacts with an antigen. The constant region is located proximal to the cell membrane. The TCR can also comprise a transmembrane region and a short cytoplasmic tail. As used herein, the term “constant region” includes the transmembrane region and cytoplasmic tail, as well as (when present) the traditional “constant region.”

[0091] The variable domain can be further subdivided into hypervariable regions, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved, termed framework regions (FRs). Each alpha chain variable domain and beta chain variable domain comprises three CDRs and four FRs: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Each variable domain comprises a binding domain that interacts with an antigen. While 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 the HLA molecule.

[0092] The term "TCR" also includes, unless otherwise indicated, antigen-binding fragments or antigen-binding portions of any of the TCRs disclosed herein, and includes monovalent and bivalent fragments or portions, as well as single chain TCRs, unless otherwise indicated. The term "TCR" is not limited to naturally occurring TCRs that are bound to the surface of a T cell. As used herein, the term "TCR" further refers to a TCR expressed on the surface of a cell other than a T cell (e.g., a cell that naturally expresses or is modified to express CD4 as described herein) or a TCR that is not contained within a cell membrane (e.g., an isolated TCR or a soluble TCR) as described herein.

[0093] An "antigen-binding molecule," "portion of a TCR," or "TCR fragment" refers to any portion of a TCR that is less than the whole. An antigen-binding molecule can include antigenic CDRs.

[0094] An "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, an "epitope" refers to a portion of a polypeptide that elicits an immune response or is capable of being bound by a TCR. The immune response can involve the production of antibodies or the activation of specific immune-competent cells, or both. Those skilled in the art will readily understand that any macromolecule, including virtually all proteins or peptides, can serve as an antigen. An antigen and / or epitope can be expressed endogenously, i.e., by genomic DNA, or can be expressed recombinantly. An antigen and / or epitope can be specific to a particular tissue, such as a cancer cell, or it can be widely expressed. Furthermore, fragments of larger molecules can serve as antigens. In an aspect, the antigen is a tumor antigen. An epitope can exist within a longer polypeptide (e.g., a protein), or an epitope can exist as a fragment of a longer polypeptide. In some aspects, an epitope is complexed with a major histocompatibility complex (MHC) (also referred to herein as complexed with an HLA molecule (e.g., an HLA class 1 molecule)).

[0095] The term "autologous" refers to any material derived from the same individual into which the material is later reintroduced. For example, autologous T cell therapy includes administration of T cells to a subject that are isolated from the same subject. The term "allogeneic" refers to any material derived from one individual that is later introduced into another individual of the same species. For example, allogeneic T cell transplantation includes administration of T cells to a subject that are obtained from a donor other than the subject.

[0096] "Cancer" refers to a broad group of various disorders characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and growth results in malignant tumors that invade neighboring tissues and can also metastasize to distant parts of the body through the lymphatic system or bloodstream. "Cancer" or "cancerous tissue" can include a tumor. Examples of cancers that can be treated by the methods of the present application include, but are not limited to, cancers of the immune system, including lymphomas, leukemias, and other white blood cell malignancies. In some aspects, the methods of the present application can be used to reduce tumor size of a tumor derived from, for example, 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 ureter cancer, renal pelvis cancer, glioma, squamous cell carcinoma, and combinations of said cancers. A particular cancer can be responsive to chemotherapy or radiation therapy, or the cancer can be refractory. Refractory cancer refers to a cancer that cannot be treated with surgical intervention, and the cancer is initially unresponsive or becomes unresponsive over time to chemotherapy or radiation therapy.

[0097] As used herein, the term "progression-free survival" can be abbreviated as PFS, and refers to the time from the treatment date to the date of disease progression according to the revised IWG response criteria for malignant lymphoma or death from any cause.

[0098] The term "overall survival" can be abbreviated as OS, and is defined as the time from the treatment date to the date of death.

[0099] As used herein, the term "infection" refers to any type of invasion of a foreign substance into one or more tissues of the body. The term "infection" includes, but is not limited to, infections caused by viruses (including viroids and prions), bacteria, fungi, parasites, and any combination thereof.

[0100] 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 reject tumors and cells infected by viruses. It acts through the process of apoptosis or programmed cell death. They are called "natural killers" because they do not require activation to kill cells. 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 special organ of the immune system, primarily responsible for the maturation of T cells. There are six types of T cells, namely: helper T cells (e.g., CD4+ cells), cytotoxic T cells (also known as TC, cytotoxic T lymphocytes, CTLs, T killer cells, cytolytic T cells, CD8+ T cells, or killer T cells), memory T cells ((i) stem memory T cells, which, like naive cells, are CD45RO-, CCR7+, CD45RA+, CD62L+ (L-selectin), CD27+, CD28+, and IL-7Ra+, but they also express large amounts of CD95, IL-2Rβ, CXCR3, and LFA-1, and show many functional attributes unique to memory cells), (ii) central memory T cells express L-selectin and CCR7, they secrete IL-2 but not IFNy or IL-4, and (iii) effector memory T cells, however, do not express L-selectin or CCR7 but produce effector cytokines such as IFNy and IL-4), regulatory T cells (Tregs, suppressor T cells, or CD4+CD25+ regulatory T cells), natural killer T cells (NKT), and gamma delta T cells. B cells, on the other hand, play a major role in humoral immunity (with antibody involvement). B cells manufacture antibodies and antigens, and function as antigen-presenting cells (APCs), and upon activation by antigen interaction, transform into memory B cells. In mammals, immature B cells are formed in the bone marrow, which is where the name originates. SCM CM EM

[0101] ​​​The terms“modified” and“mutated” when used herein in reference to a nucleotide or amino acid sequence refer to a change in the sequence relative to a wild-type sequence or a particular reference sequence. Unless otherwise specified, the terms“modified” and“mutated” do not require steps in the process used to make the modified or mutated sequence (e.g., a modified beta chain sequence). Rather, these terms indicate that the modified or mutated sequence has a variation relative to a reference sequence (e.g., a wild-type sequence). For example, a DP beta chain comprising a substitution mutation at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 does not require that the wild-type DP beta chain has been physically altered to arrive at the DP beta chain; rather, when properly aligned, the DP beta chain comprises an amino acid residue at the position (residue 112) that is different from the amino acid residue at the corresponding position in the wild-type or reference DP beta chain.

[0102] As used herein, the term“any amino acid” refers to any known amino acid. An amino acid is an organic compound that contains (i) an amine (-NH2) functional group, (ii) a carboxyl (-COOH)_ functional group, and (iii) a side chain (R group), where the side chain is specific to each amino acid. This includes, but is not limited to, any naturally occurring amino acid as well as any modification and variant 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 (lie; I), leucine (Leu; L), methionine (Met; M), phenylalanine (Phe; F), valine (Val; V), tryptophan (Trp; W), tyrosine (Tyr; Y). Tryptophan (Trp; W), tyrosine (Tyr; Y), and methionine (Met; M) can also be classified as polar and / or amphipathic amino acids, as these amino acids are often found on the surface of a protein or lipid membrane. Additional amino acids include cysteine (Cys; C), selenocysteine (Sec; U), glycine (Gly; G), and proline (Pro; P).

[0103] As used herein, “at a position corresponding to” is used as a means to identify a particular amino acid residue (e.g., a particular amino acid position) in a polynucleotide or a particular nucleic acid (e.g., a particular nucleic acid position) in a polypeptide. The position can be determined by aligning the sequence in question with a reference sequence properly. One of skill 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).

[0104] The term “genetically engineered” or “engineered” refers to a method of modifying the genome of a cell, including but not limited to deletion of a coding or non-coding region or portion thereof, or insertion of a coding region or portion thereof. In some aspects, the modified cell is a lymphocyte, such as a T cell expressing CD4 or a modified cell, which can be obtained from a patient or a donor. The cell can be modified to express an exogenous construct, such as a T cell receptor (TCR) as disclosed herein, which is incorporated into the cell genome. In some aspects, the cell is modified to express CD4.

[0105] “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 produced by any of these cells or the liver (including Abs, cytokines, and complement) that result in the selective targeting, binding, impairment, destruction, and / or elimination of invading pathogens, cells or tissues infected with a pathogen, cancer cells or other abnormal cells, or (in the case of autoimmune or pathological inflammation) normal body cells or tissues in a vertebrate.

[0106] The term “immunotherapy” refers to the treatment of a subject having a disease or at risk of contracting a disease or suffering a recurrence of a disease by a method that includes inducing, enhancing, suppressing, or otherwise modifying an immune response. Examples of immunotherapy include, but are not limited to, T cell therapy. T cell therapy can include adoptive T cell therapy, tumor infiltrating lymphocyte (TIL) immunotherapy, autologous cell therapy, engineered autologous cell therapy (eACT), and allogeneic T cell transplantation.

[0107] The cells used in the immunotherapy described herein can be from any source known in the art. For example, T cells can be differentiated in vitro from a population of hematopoietic stem cells, or the 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 a site of infection, ascites, pleural effusion, spleen tissue, and tumors. In addition, the T cells can be derived from one or more T cell lines available in the art. T cells can also be obtained from unit blood collected from a subject using any number of techniques known to those of skill in the art, such as FICOLL TM separation and / or apheresis). Additional methods of isolating T cells for use in T cell therapy are disclosed in U.S. Patent Publication No. 2013 / 0287748, which is incorporated by reference herein in its entirety. The immunotherapy can also include administering to the subject a modified cell, wherein the modified cell expresses a CD4 and a TCR disclosed herein. In some aspects, the modified cell is not a T cell.

[0108] As used herein, a“patient” includes any human who has cancer (e.g., lymphoma or leukemia). The terms“subject” and“patient” are used interchangeably herein.

[0109] The terms“peptide,”“polypeptide,” and“protein” are used interchangeably and refer to a compound composed of amino acid residues connected to one another 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 of a sequence that can make up a protein or peptide. A polypeptide includes any peptide or protein comprising two or more amino acids connected to one another by peptide bonds. As used herein, the terms refer to both short chains, e.g., often referred to in the art as peptides, oligopeptides and oligomers, as well as longer chains often referred to in the art as proteins, of which there are many types. “Polypeptide” includes, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, and the like. Polypeptides include natural, recombinant, synthetic, or combinations thereof.

[0110] As used herein, "stimulation" refers to a primary response induced by the binding of a stimulatory molecule to its cognate ligand, wherein the binding mediates a signal transduction event. A "stimulatory molecule" is a molecule on a T cell, such as the T cell receptor (TCR) / CD4 complex, that specifically binds to a cognate stimulatory ligand present on an antigen presenting cell. A "stimulatory ligand" is a molecule that, when present on an antigen presenting cell (e.g., aAPC, dendritic cell, B cell, etc.) can specifically bind to a stimulatory molecule on a T cell, thereby mediating a primary response by the T cell, including but not limited to activation, initiation of an immune response, proliferation, etc. Stimulatory ligands include, but are not limited to, MHC class II molecules loaded with peptide, anti-CD4 antibodies, anti-CD28 antibodies, anti-CD2 antibodies, and anti-CD3 antibodies.

[0111] "treatment" or "treating" a subject refers to any type of intervention or process performed on the subject, or administration of an active agent to the subject, with the objective of reversing, alleviating, improving, inhibiting, slowing down or preventing the onset of the symptoms, complications or condition, or the progression, development, severity or recurrence of a disease-related biochemical indicia. In one aspect, "treatment" or "treating" includes partial alleviation. In another aspect, "treatment" or "treating" includes complete alleviation.

[0112] Use of alternatives (e.g., "or") should be understood as meaning one, the other, or both or any combination thereof. As used herein, the indefinite article "a" or "an" should be understood to mean "one or more" of any described or recited component.

[0113] The terms "about" or "substantially" refer to values or compositions that are within an acceptable error range for the specific value or composition determined by one of ordinary skill in the art, which is between about 1 and 10% of the value or composition. For example, "about" or "substantially" can refer to within 1 or more than 1 standard deviation, per the practice in the art. Alternatively, "about" or "substantially" can mean up to 10% (i.e., ±10%) of the stated value. For example, about 3 mg can include any number between 2.7 mg and 3.3 mg (10%). Furthermore, particularly with respect to biological systems or processes, the terms can mean up to an order of magnitude or up to five-fold of a value. When a particular value or composition is provided in the application and claims, unless otherwise stated the intent is that "about" or "substantially" be assumed in the recitation of the value or composition to capture values or compositions within an acceptable error range.

[0114] Any concentration range, percentage range, ratio range, or integer range is to be understood to include also any integer or fraction within that range.

[0115] Various aspects of the application are described in more detail in the following subsections.

[0116] II. Compositions of the Disclosure

[0117] The present disclosure relates to HLA class II molecules with enhanced CD4 binding. Certain aspects of the disclosure relate to HLA class II molecules comprising a beta chain, wherein the beta chain comprises one or more mutations. In certain aspects, the one or more mutations in the beta chain increase the affinity of the beta chain for CD4. In certain aspects, the beta chain is an HLA-DP (“DP”) beta chain.

[0118] II. A. MHC class II molecules

[0119] 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 that specialize in presenting antigenic peptides to T cell receptors (TCRs) on T cells. (See also Overview of the Immune System). MHC molecules that present antigens (Ag) are divided into two broad classes: class I MHC molecules and class II MHC molecules.

[0120] Class II MHC molecules exist as transmembrane glycoproteins on the surface of professional antigen-presenting cells (APCs). The complete class II molecule is composed of an alpha chain and a beta chain. Three loci in the HLA complex encode MHC class II proteins: HLA-DP, HLA-DQ, and HLA-DR. T cells that express the CD4 molecule react with class II MHC molecules. These lymphocytes generally have effector and helper functions and activate responses to eliminate self cells infected with intracellular pathogens or to destroy extracellular parasites and help 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 nonpolymorphic portions of the alpha-2 and beta-2 domains of the alpha and beta chains of MHC class II molecules, respectively).

[0121] In some aspects, the HLA class II alpha and beta chains are selected from HLA-DP, HLA-DQ, and HLA-DR alleles. In certain aspects, the HLA class II beta chain is an HLA-DP allele. In certain aspects, the HLA class II alpha chain is an HLA-DP allele.

[0122] Many HLA-DP alleles are known in the art, and any known allele can be used in the present disclosure. Examples of HLA-DP alpha and beta chain alleles are shown in Table 1. An updated list of HLA alleles is available at hla.alleles.org / (last accessed February 27, 2019).

[0123] II.A.1. MHC Class II Beta Chain

[0124] In certain aspects, the HLA Class II molecule comprises a DP beta chain, wherein the DP beta chain comprises an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1. Any amino acid other than leucine can be present at a position corresponding to amino acid residue 112 of SEQ ID NO: 1. In some aspects, the amino acid other than leucine is an amino acid comprising a hydrophobic side chain. In certain aspects, the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is an amino acid selected from the group consisting of alanine, valine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan. In certain aspects, the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is alanine. In certain aspects, the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is valine. In certain aspects, the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is isoleucine. In certain aspects, the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is methionine. In certain aspects, the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is phenylalanine. In certain aspects, the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is tyrosine. In certain aspects, the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is tryptophan.

[0125] In some embodiments, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is composed of more than one amino acid, e.g., two amino acids, three amino acids, four amino acids, five amino acids, or more. In some aspects, at least one of the more than one amino acids comprises a hydrophobic side chain. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is composed of a series, e.g., at least 2, at least 3, at least 4, or at least 5 amino acids, wherein each of the series of amino acids comprises a hydrophobic side chain.

[0126] In certain aspects, the HLA class II molecule comprises a DP beta chain, wherein the DP beta chain comprises an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1. Any amino acid other than valine can be present at a position corresponding to amino acid residue 141 of SEQ ID NO: 1. In some aspects, the amino acid other than valine is an amino acid comprising a hydrophobic side chain. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is an amino acid selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is alanine. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is isoleucine. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is leucine. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is methionine. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is phenylalanine. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is tyrosine. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is tryptophan.

[0127] In some aspects, the amino acid other than valine at the position corresponding to amino acid residue 141 of SEQ ID NO: 1 is composed of more than one amino acid, e.g., two amino acids, three amino acids, four amino acids, five amino acids, or more. In some aspects, at least one of the more than one amino acids comprises a hydrophobic side chain. In certain aspects, the amino acid other than valine at the position corresponding to amino acid residue 141 of SEQ ID NO: 1 is composed of a series, e.g., at least 2, at least 3, at least 4, or at least 5 amino acids, wherein each of the series of amino acids comprises a hydrophobic side chain.

[0128] In certain aspects of the disclosure, the MHC class II molecule comprises a DP b chain containing more than one substitution mutation relative to a wild-type DP b chain. In certain aspects, the DP b chain comprises at least two mutations, at least three mutations, at least four mutations, at least five mutations, at least six mutations, at least seven mutations, at least eight mutations, at least nine mutations, or at least ten mutations relative to a wild-type DP b chain.

[0129] In certain aspects, the DP b chain comprises an amino acid other than leucine at the position corresponding to amino acid residue 112 of SEQ ID NO: 1 and an amino acid other than valine at the position corresponding to amino acid residue 141 of SEQ ID NO: 1. In some aspects, each of (i) the amino acid other than leucine at the position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) the amino acid other than valine at the position corresponding to amino acid residue 141 of SEQ ID NO: 1, or the amino acid other than leucine at the position corresponding to amino acid residue 112 of SEQ ID NO: 1 and the amino acid other than valine at the position corresponding to amino acid residue 141 of SEQ ID NO: 1 is an amino acid comprising a hydrophobic side chain. In some aspects, (i) the amino acid other than leucine at the position corresponding to amino acid residue 112 of SEQ ID NO: 1 is selected from the group consisting of alanine, valine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan; and (ii) the amino acid other than valine at the position corresponding to amino acid residue 141 of SEQ ID NO: 1 is selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan.

[0130] In some aspects, (i) the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is tryptophan; and (ii) the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In some aspects, (i) the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is selected from the group consisting of alanine, valine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan; and (ii) the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is methionine. In some aspects, (i) the amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 is tryptophan; and (ii) the amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1 is methionine.

[0131] In certain aspects, the DP beta chain further comprises an amino acid other than valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1. In some aspects, the amino acid other than valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1 is selected from the group consisting of alanine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan. In certain aspects, the amino acid other than valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1 is methionine.

[0132] In certain aspects, the DP beta chain further comprises an amino acid other than methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1. In some aspects, the amino acid other than methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1 is selected from the group consisting of alanine, valine, isoleucine, methionine, phenylalanine, tyrosine, and tryptophan. In certain aspects, the amino acid other than methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1 is isoleucine.

[0133] In some aspects, the DP beta chain comprises (i) an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, and (ii) an amino acid other than valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1. In some aspects, the DP beta chain comprises (i) an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, and (ii) an amino acid other than methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0134] In some aspects, the DP beta chain comprises (i) tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, and (ii) methionine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1. In some aspects, the DP beta chain comprises (i) tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, and (ii) isoleucine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0135] In some aspects, the DP beta chain comprises (i) an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, and (ii) an amino acid other than valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1. In some aspects, the DP beta chain comprises (i) an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, and (ii) an amino acid other than methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0136] In some aspects, the DP beta chain comprises (i) methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, and (ii) an amino acid other than valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1. In some aspects, the DP beta chain comprises (i) methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, and (ii) isoleucine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0137] In some aspects, the DP beta chain comprises (i) an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, (iii) an amino acid other than valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1, and (iv) an amino acid other than methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0138] In some aspects, the DP beta chain comprises (i) tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, (iii) methionine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1, and (iv) isoleucine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0139] In some aspects, the DP beta chain comprises valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1. In some aspects, the DP beta chain comprises methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1. In some aspects, the DP beta chain comprises (i) an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, and (iii) valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1. In some aspects, the DP beta chain comprises (i) an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, and (iii) methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1. In some aspects, the DP beta chain comprises (i) an amino acid other than leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) an amino acid other than valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, (iii) valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1, and (iv) methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0140] In some aspects, the DP β chain comprises (i) a tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) a methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, and (iii) a valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1. In some aspects, the DP β chain comprises (i) a tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) a methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, and (iii) a methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1. In some aspects, the DP β chain comprises (i) a tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) a methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, (iii) a valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1, and (iv) a methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0141] In some aspects, the DP β chain comprises (i) a tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) a methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, (iii) a methionine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1, and (iv) an isoleucine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1.

[0142] In certain aspects, a DP β chain described herein has increased affinity for a CD4 protein compared to a reference HLA class II molecule. In some aspects, the reference HLA class II molecule is an HLA class II molecule having a wild-type DP β chain. In some aspects, the reference HLA class II molecule is an HLA class II molecule having a DP β chain comprising (i) a leucine at a position corresponding to amino acid residue 112 of SEQ ID NO: 1 and / or (ii) a valine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1.

[0143] In some aspects, the increased affinity for CD4 is at least about 1.5-fold, at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least about 8-fold, at least about 9-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, at least about 30-fold, at least about 35-fold, at least about 40-fold, at least about 45-fold, at least about 50-fold, at least about 75-fold, at least about 100-fold, at least about 200-fold, at least about 300-fold, at least about 400-fold, at least about 500-fold, at least about 1000-fold, at least about 1500-fold, at least about 2000-fold, at least about 2500-fold, at least about 3000-fold, at least about 3500-fold, at least about 4000-fold, at least about 4500-fold, or at least about 4000-fold of the reference HLA class II molecule's affinity for CD4.

[0144] In some aspects, the increased affinity for CD4 is at least about 1.5-fold to at least about 5000-fold, 1.5-fold to at least about 4000-fold, 1.5-fold to at least about 3000-fold, 1.5-fold to at least about 2000-fold, 1.5-fold to at least about 1000-fold, 10-fold to at least about 5000-fold, 10-fold to at least about 4000-fold, 10-fold to at least about 3000-fold, 10-fold to at least about 2000-fold, 10-fold to at least about 1000-fold, 10-fold to at least about 900-fold, 10-fold to at least about 800-fold, 10-fold to at least about 700-fold, 10-fold to at least about 600-fold, 10-fold to at least about 500-fold, 10-fold to at least about 400-fold, 10-fold to at least about 300-fold, 10-fold to at least about 200-fold, 10-fold to at least about 100-fold, 100-fold to at least about 5000-fold, 100-fold to at least about 4000-fold, 100-fold to at least about 3000-fold, 100-fold to at least about 2000-fold, 100-fold to at least about 1000-fold, 100-fold to at least about 900-fold, 100-fold to at least about 800-fold, 100-fold to at least about 700-fold, 100-fold to at least about 600-fold, 100-fold to at least about 500-fold, 100-fold to at least about 400-fold, 100-fold to at least about 300-fold, or 100-fold to at least about 200-fold of the reference HLA class II molecule's affinity for CD4.

[0145] In certain aspects, the DP beta chain comprises an allele selected from the group consisting of DPB1*01, DPB1*02, DPB1*03, DPB1*04, DPB1*05, DPB1*06, DPB1*08, DPB1*09, DPB1*10, DPB1*100, DPB1*101, DPB1*102, DPB1*103, DPB1*104, DPB1*105, DPB1*106, DPB1*107, DPB1*108, DPB1*109, DPB1*11, DPB1*110, DPB1*111, DPB1*112, DPB1*113, DPB1*114, DPB1*115, DPB1*116, DPB1*117, DPB1*118, DPB1*119, DPB1*120, DPB1*121, DPB1*122, DPB1*123, DPB1*124, DPB1*125, DPB1*126, DPB1*127, DPB1*128, DPB1*129, DPB1*13, DPB1*130, DPB1*131, DPB1*132, DPB1*133, DPB1*134, DPB1*135, DPB1*136, DPB1*137, DPB1*138, DPB1*139, DPB1*14, DPB1*140, DPB1*141, DPB1*142, DPB1*143, DPB1*144, DPB1*145, DPB1*146, DPB1*147, DPB1*148, DPB1*149, DPB1*15, DPB1*150, DPB1*151, DPB1*152, DPB1*153, DPB1*154, DPB1*155, DPB1*156, DPB1*157, DPB1*158, DPB1*159, DPB1*16, DPB1*160, DPB1*161, DPB1*162, DPB1*163, DPB1*164, DPB1*165, DPB1*166, DPB1*167, DPB1*168, DPB1*169, DPB1*17, DPB1*170, DPB1*171, DPB1*172, DPB1*173, DPB1*174, DPB1*175, DPB1*176, DPB1*177, DPB1*178, DPB1*179, DPB1*18, DPB1*180, DPB1*181, DPB1*182, DPB1*183, DPB1*184, DPB1*185, DPB1*186, DPB1*187, DPB1*188, DPB1*189, DPB1*19, DPB1*190, DPB1*191, DPB1*192,DPB1*193, DPB1*194, DPB1*195, DPB1*196, DPB1*197, DPB1*198, DPB1*199, DPB1*20, DPB1*200, DPB1*201, DPB1*202, DPB1*203, DPB1*204, DPB1*205, DPB1*206, DPB1*207, DPB1*208, DPB1*209, DPB1*21, DPB1*210, DPB1*211, DPB1*212, DPB1*213, DPB1*214, DPB1*215, DPB1*216, DPB1*217, DPB1*218, DPB1*219, DPB1*22, DPB1*220, DPB1*221, DPB1*222, DPB1*223, DPB1*224, DPB1*225, DPB1*226, DPB1*227, DPB1*228, DPB1*229, DPB1*23, DPB1*230, DPB1*231, DPB1*232, DPB1*233, DPB1*234, DPB1*235, DPB1*236, DPB1*237, DPB1*238, DPB1*239, DPB1*24, DPB1*240, DPB1*241, DPB1*242, DPB1*243, DPB1*244, DPB1*245, DPB1*246, DPB1*247, DPB1*248, DPB1*249, DPB1*25, DPB1*250, DPB1*251, DPB1*252, DPB1*253, DPB1*254, DPB1*255, DPB1*256, DPB1*257, DPB1*258, DPB1*259, DPB1*26, DPB1*260, DPB1*261, DPB1*262, DPB1*263, DPB1*264, DPB1*265, DPB1*266, DPB1*267, DPB1*268, DPB1*269, DPB1*27, DPB1*270, DPB1*271, DPB1*272, DPB1*273, DPB1*274, DPB1*275, DPB1*276, DPB1*277, DPB1*278, DPB1*279, DPB1*28, DPB1*280, DPB1*281, DPB1*282, DPB1*283, DPB1*284, DPB1*285, DPB1*286, DPB1*287, DPB1*288, DPB1*289, DPB1*29, DPB1*290, DPB1*291, DPB1*292, DPB1*293, DPB1*294,DPB1*295, DPB1*296, DPB1*297, DPB1*298, DPB1*299, DPB1*30, DPB1*300, DPB1*301, DPB1*302, DPB1*303, DPB1*304, DPB1*305, DPB1*306, DPB1*307, DPB1*308, DPB1*309, DPB1*31, DPB1*310, DPB1*311, DPB1*312, DPB1*313, DPB1*314, DPB1*315, DPB1*316, DPB1*317, DPB1*318, DPB1*319, DPB1*32, DPB1*320, DPB1*321, DPB1*322, DPB1*323, DPB1*324, DPB1*325, DPB1*326, DPB1*327, DPB1*328, DPB1*329, DPB1*33, DPB1*330, DPB1*331, DPB1*332, DPB1*333, DPB1*334, DPB1*335, DPB1*336, DPB1*337, DPB1*338, DPB1*339, DPB1*34, DPB1*340, DPB1*341, DPB1*342, DPB1*343, DPB1*344, DPB1*345, DPB1*346, DPB1*347, DPB1*348, DPB1*349, DPB1*35, DPB1*350, DPB1*351, DPB1*352, DPB1*353, DPB1*354, DPB1*355, DPB1*356, DPB1*357, DPB1*358, DPB1*359, DPB1*36, DPB1*360, DPB1*361, DPB1*362, DPB1*363, DPB1*364, DPB1*365, DPB1*366, DPB1*367, DPB1*368, DPB1*369, DPB1*37, DPB1*370, DPB1*371, DPB1*372, DPB1*373, DPB1*374, DPB1*375, DPB1*376, DPB1*377, DPB1*378, DPB1*379, DPB1*38, DPB1*380, DPB1*381, DPB1*382, DPB1*383, DPB1*384, DPB1*385, DPB1*386, DPB1*387, DPB1*388, DPB1*389, DPB1*39, DPB1*390, DPB1*391, DPB1*392, DPB1*393, DPB1*394, DPB1*395, DPB1*396,DPB1*397, DPB1*398, DPB1*399, DPB1*40, DPB1*400, DPB1*401, DPB1*402, DPB1*403, DPB1*404, DPB1*405, DPB1*406, DPB1*407, DPB1*408, DPB1*409, DPB1*41, DPB1*410, DPB1*411, DPB1*412, DPB1*413, DPB1*414, DPB1*415, DPB1*416, DPB1*417, DPB1*418, DPB1*419, DPB1*420, DPB1*421, DPB1*422, DPB1*423, DPB1*424, DPB1*425, DPB1*426, DPB1*427, DPB1*428, DPB1*429, DPB1*430, DPB1*431, DPB1*432, DPB1*433, DPB1*434, DPB1*435, DPB1*436, DPB1*437, DPB1*438, DPB1*439, DPB1*44, DPB1*440, DPB1*441, DPB1*442, DPB1*443, DPB1*444, DPB1*445, DPB1*446, DPB1*447, DPB1*448, DPB1*449, DPB1*45, DPB1*450, DPB1*451, DPB1*452, DPB1*453, DPB1*454, DPB1*455, DPB1*456, DPB1*457, DPB1*458, DPB1*459, DPB1*46, DPB1*460, DPB1*461, DPB1*462, DPB1*463, DPB1*464, DPB1*465, DPB1*466, DPB1*467, DPB1*468, DPB1*469, DPB1*47, DPB1*470, DPB1*471, DPB1*472, DPB1*473, DPB1*474, DPB1*475, DPB1*476, DPB1*477, DPB1*478, DPB1*479, DPB1*48, DPB1*480, DPB1*481, DPB1*482, DPB1*483, DPB1*484, DPB1*485, DPB1*486, DPB1*487, DPB1*488, DPB1*489, DPB1*49, DPB1*490, DPB1*491, DPB1*492, DPB1*493, DPB1*494, DPB1*495, DPB1*496, DPB1*497, DPB1*498, DPB1*499, DPB1*50,DPB1*500, DPB1*501, DPB1*502, DPB1*503, DPB1*504, DPB1*505, DPB1*506, DPB1*507, DPB1*508, DPB1*509, DPB1*51, DPB1*510, DPB1*511, DPB1*512, DPB1*513, DPB1*514, DPB1*515, DPB1*516, DPB1*517, DPB1*518, DPB1*519, DPB1*52, DPB1*520, DPB1*521, DPB1*522, DPB1*523, DPB1*524, DPB1*525, DPB1*526, DPB1*527, DPB1*528, DPB1*529, DPB1*53, DPB1*530, DPB1*531, DPB1*532, DPB1*533, DPB1*534, DPB1*535, DPB1*536, DPB1*537, DPB1*538, DPB1*539, DPB1*54, DPB1*540, DPB1*541, DPB1*542, DPB1*543, DPB1*544, DPB1*545, DPB1*546, DPB1*547, DPB1*548, DPB1*549, DPB1*55, DPB1*550, DPB1*551, DPB1*552, DPB1*553, DPB1*554, DPB1*555, DPB1*556, DPB1*557, DPB1*558, DPB1*559, DPB1*56, DPB1*560, DPB1*561, DPB1*562, DPB1*563, DPB1*564, DPB1*565, DPB1*566, DPB1*567, DPB1*568, DPB1*569, DPB1*57, DPB1*570, DPB1*571, DPB1*572, DPB1*573, DPB1*574, DPB1*575, DPB1*576, DPB1*577, DPB1*578, DPB1*579, DPB1*58, DPB1*580, DPB1*581, DPB1*582, DPB1*583, DPB1*584, DPB1*585, DPB1*586, DPB1*587, DPB1*588, DPB1*589, DPB1*59, DPB1*590, DPB1*591, DPB1*592, DPB1*593, DPB1*594, DPB1*595, DPB1*596, DPB1*597, DPB1*598, DPB1*599, DPB1*60, DPB1*600, DPB1*601,DPB1*602, DPB1*603, DPB1*604, DPB1*605, DPB1*606, DPB1*607, DPB1*608, DPB1*609, DPB1*61, DPB1*610, DPB1*611, DPB1*612, DPB1*613, DPB1*614, DPB1*615, DPB1*616, DPB1*617, DPB1*618, DPB1*619, DPB1*62, DPB1*620, DPB1*621, DPB1*622, DPB1*623, DPB1*624, DPB1*625, DPB1*626, DPB1*627, DPB1*628, DPB1*629, DPB1*63, DPB1*630, DPB1*631, DPB1*632, DPB1*633, DPB1*634, DPB1*635, DPB1*636, DPB1*637, DPB1*638, DPB1*639, DPB1*64, DPB1*640, DPB1*641, DPB1*642, DPB1*643, DPB1*644, DPB1*645, DPB1*646, DPB1*647, DPB1*648, DPB1*649, DPB1*65, DPB1*650, DPB1*651, DPB1*652, DPB1*653, DPB1*654, DPB1*655, DPB1*656, DPB1*657, DPB1*658, DPB1*659, DPB1*66, DPB1*660, DPB1*661, DPB1*662, DPB1*663, DPB1*664, DPB1*665, DPB1*666, DPB1*667, DPB1*668, DPB1*669, DPB1*67, DPB1*670, DPB1*671, DPB1*672, DPB1*673, DPB1*674, DPB1*675, DPB1*676, DPB1*677, DPB1*678, DPB1*679, DPB1*68, DPB1*680, DPB1*681, DPB1*682, DPB1*683, DPB1*684, DPB1*685, DPB1*686, DPB1*687, DPB1*688, DPB1*689, DPB1*69, DPB1*690, DPB1*691, DPB1*692, DPB1*693, DPB1*694, DPB1*695, DPB1*696, DPB1*697, DPB1*698, DPB1*699, DPB1*70, DPB1*700, DPB1*701, DPB1*702, DPB1*703,DPB1*704, DPB1*705, DPB1*706, DPB1*707, DPB1*708, DPB1*709, DPB1*71, DPB1*710, DPB1*711, DPB1*712, DPB1*713, DPB1*714, DPB1*715, DPB1*716, DPB1*717, DPB1*718, DPB1*719, DPB1*72, DPB1*720, DPB1*721, DPB1*722, DPB1*723, DPB1*724, DPB1*725, DPB1*726, DPB1*727, DPB1*728, DPB1*729, DPB1*73, DPB1*730, DPB1*731, DPB1*732, DPB1*733, DPB1*734, DPB1*735, DPB1*736, DPB1*737, DPB1*738, DPB1*739, DPB1*74, DPB1*740, DPB1*741, DPB1*742, DPB1*743, DPB1*744, DPB1*745, DPB1*746, DPB1*747, DPB1*748, DPB1*749, DPB1*75, DPB1*750, DPB1*751, DPB1*752, DPB1*753, DPB1*754, DPB1*755, DPB1*756, DPB1*757, DPB1*758, DPB1*759, DPB1*76, DPB1*760, DPB1*761, DPB1*762, DPB1*763, DPB1*764, DPB1*765, DPB1*766, DPB1*767, DPB1*768, DPB1*769, DPB1*77, DPB1*770, DPB1*771, DPB1*772, DPB1*773, DPB1*774, DPB1*775, DPB1*776, DPB1*777, DPB1*778, DPB1*779, DPB1*78, DPB1*780, DPB1*781, DPB1*782, DPB1*783, DPB1*784, DPB1*785, DPB1*786, DPB1*787, DPB1*788, DPB1*789, DPB1*79, DPB1*790, DPB1*791, DPB1*792, DPB1*794, DPB1*795, DPB1*796, DPB1*797, DPB1*798, DPB1*799, DPB1*80, DPB1*800, DPB1*801, DPB1*802, DPB1*803, DPB1*804, DPB1*805, DPB1*806,DPB1*807, DPB1*808, DPB1*809, DPB1*81, DPB1*810, DPB1*811, DPB1*812, DPB1*813, DPB1*814, DPB1*815, DPB1*816, DPB1*817, DPB1*818, DPB1*819, DPB1*82, DPB1*820, DPB1*821, DPB1*822, DPB1*823, DPB1*824, DPB1*825, DPB1*826, DPB1*827, DPB1*828, DPB1*829, DPB1*83, DPB1*830, DPB1*831, DPB1*832, DPB1*833, DPB1*834, DPB1*835, DPB1*836, DPB1*837, DPB1*838, DPB1*839, DPB1*84, DPB1*840, DPB1*841, DPB1*842, DPB1*843, DPB1*844, DPB1*845, DPB1*846, DPB1*847, DPB1*848, DPB1*849, DPB1*85, DPB1*850, DPB1*851, DPB1*852, DPB1*853, DPB1*854, DPB1*855, DPB1*856, DPB1*857, DPB1*858, DPB1*859, DPB1*86, DPB1*860, DPB1*861, DPB1*862, DPB1*863, DPB1*864, DPB1*865, DPB1*866, DPB1*867, DPB1*868, DPB1*869, DPB1*87, DPB1*870, DPB1*871, DPB1*872, DPB1*873, DPB1*874, DPB1*875, DPB1*876, DPB1*877, DPB1*878, DPB1*879, DPB1*88, DPB1*880, DPB1*881, DPB1*882, DPB1*883, DPB1*884, DPB1*885, DPB1*886, DPB1*887, DPB1*888, DPB1*889, DPB1*89, DPB1*890, DPB1*891, DPB1*892, DPB1*893, DPB1*894, DPB1*895, DPB1*896, DPB1*897, DPB1*898, DPB1*899, DPB1*90, DPB1*900, DPB1*901, DPB1*902, DPB1*903, DPB1*904, DPB1*905, DPB1*906, DPB1*907, DPB1*908,DPB1*909, DPB1*91, DPB1*910, DPB1*911, DPB1*912, DPB1*913, DPB1*914, DPB1*915, DPB1*916, DPB1*917, DPB1*918, DPB1*919, DPB1*92, DPB1*920, DPB1*921, DPB1*922, DPB1*923, DPB1*924, DPB1*925, DPB1*926, DPB1*927, DPB1*928, DPB1*929, DPB1*93, DPB1*930, DPB1*931, DPB1*932, DPB1*933, DPB1*934, DPB1*935, DPB1*936, DPB1*937, DPB1*938, DPB1*939, DPB1*94, DPB1*940, DPB1*941, DPB1*942, DPB1*943, DPB1*944, DPB1*945, DPB1*946, DPB1*947, DPB1*948, DPB1*949, DPB1*95, DPB1*950, DPB1*951, DPB1*952, DPB1*953, DPB1*954, DPB1*955, DPB1*956, DPB1*957, DPB1*958, DPB1*959, DPB1*96, DPB1*960, DPB1*961, DPB1*962, DPB1*963, DPB1*964, DPB1*965, DPB1*97, DPB1*98, and DPB1*99. In some aspects, the DP beta chain comprises an HLA-DPB1*01, HLA-DPB1*02, HLA-DPB1*03, HLA-DPB1*04, HLA-DPB1*05, HLA-DPB1*06, HLA-DPB1*08, or HLA-DPB1*09 allele. In certain aspects, the DP beta chain comprises an HLA-DPB1*04 allele. In particular aspects, the DP beta chain comprises an HLA-DPB1*04:01 allele.

[0146] In certain aspects, the MHC class II molecule comprises a DP beta chain comprising 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 to SEQ ID NO: 3, wherein the DP beta chain comprises a tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, and wherein the DP beta chain comprises a methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1. In certain aspects, the MHC class II molecule comprises a DP beta chain comprising 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 to SEQ ID NO: 3, wherein the DP beta chain comprises (i) a tryptophan at a position corresponding to amino acid residue 112 of SEQ ID NO: 1, (ii) a methionine at a position corresponding to amino acid residue 141 of SEQ ID NO: 1, (iii) a valine at a position corresponding to amino acid residue 114 of SEQ ID NO: 1, and (iv) a methionine at a position corresponding to amino acid residue 158 of SEQ ID NO: 1. In certain aspects, the MHC class II molecule comprises a DP beta chain comprising the amino acid sequence set forth in SEQ ID NO: 3.

[0147] II.A.2. MHC class II alpha chain

[0148] In some aspects of the disclosure, the MHC class II molecule further comprises an alpha chain. In some aspects, the alpha chain is a wild-type alpha chain. In some aspects, the alpha chain is a DP alpha chain. Any DP alpha chain can be used in the compositions and methods of the disclosure. In some aspects, the DP alpha chain comprises an HLA-DPA1*01, HLA-DPA1*02, HLA-DPA1*03, or HLA-DPA1*04 allele. In certain aspects, the DP alpha chain comprises an HLA-DPA1*01 allele. In certain aspects, the DP alpha chain comprises an HLA-DPA1*02 allele. In certain aspects, the DP alpha chain comprises an HLA-DPA1*03 allele. In certain aspects, the DP alpha chain comprises an HLA-DPA1*04 allele.

[0149] In certain aspects, the DP a chain is selected from DPA1*01:03:01:01, DPA1*01:03:01:02, DPA1*01:03:01:03, DPA1*01:03:01:04, DPA1*01:03:01:05, DPA1*01:03:01:06, DPA1*01:03:01:07, DPA1*01:03:01:08, DPA1*01:03:01:09, DPA1*01:03:01:10, DPA1*01:03:01:11, DPA1*01:03:01:12, DPA1*01:03:01:13, DPA1*01:03:01:14, DPA1*01:03:01:15, DPA1*01:03:01:16, DPA1*01:03:01:17, DPA1*01:03:01:18Q, DPA1*01:03:01:19, DPA1*01:03:01:20, DPA1*01:03:01:21, DPA1*01:03:01:22, DPA1*01:03:01:23, DPA1*01:03:02, DPA1*01:03:03, DPA1*01:03:04, DPA1*01:03:05, DPA1*01:03:06, DPA1*01:03:07, DPA1*01:03:08, DPA1*01:03:09, DPA1*01:04, DPA1*01:05, DPA1*01:06:01, DPA1*01:06:02, DPA1*01:07, DPA1*01:08, DPA1*01:09, DPA1*01:10, DPA1*01:11, DPA1*01:12, DPA1*01:13, DPA1*01:14, DPA1*01:15, DPA1*01:16, DPA1*01:17, DPA1*01:18, DPA1*01:19, DPA1*02:01:01:01, DPA1*02:01:01:02, DPA1*02:01:01:03, DPA1*02:01:01:04, DPA1*02:01:01:05, DPA1*02:01:01:06, DPA1*02:01:01:07, DPA1*02:01:01:08, DPA1*02:01:01:09, DPA1*02:01:01:10, DPA1*02:01:01:11, DPA1*02:01:02:01, DPA1*02:01:02:02, DPA1*02:01:03, DPA1*02:01:04, DPA1*02:01:05, DPA1*02:01:06, DPA1*02:01:07,DPA1*02:01:08:01, DPA1*02:01:08:02, DPA1*02:02:02:01, DPA1*02:02:02:02, DPA1*02:02:02:03, DPA1*02:02:02:04, DPA1*02:02:02:05, DPA1*02:02:03, DPA1*02:02:04, DPA1*02:02:05, DPA1*02:02:06, DPA1*02:03, DPA1*02:04, DPA1*02:05, DPA1*02:06, DPA1*02:07:01:01, DPA1*02:07:01:02, DPA1*02:07:01:03, DPA1*02:08, DPA1*02:09, DPA1*02:10, DPA1*02:11, DPA1*02:12, DPA1*02:13N, DPA1*02:14, DPA1*02:15, DPA1*02:16, DPA1*03:01:01:01, DPA1*03:01:01:02, DPA1*03:01:01:03, DPA1*03:01:01:04, DPA1*03:01:01:05, DPA1*03:01:02, DPA1*03:02, DPA1*03:03, DPA1*03:04, DPA1*04:01:01:01, DPA1*04:01:01:02, DPA1*04:01:01:03, DPA1*04:02, or any combination thereof.

[0150] II.A.3. Signal Peptide

[0151] In some aspects, the DP beta chain and / or DP alpha chain further comprises a signal peptide. Any signal peptide known in the art can be used in the compositions and methods disclosed herein. In some aspects, the DP beta chain signal peptide is the same as the DP alpha signal peptide. In some aspects, the DP beta chain signal peptide is different from the DP alpha signal peptide.

[0152] In some aspects, the signal peptide is derived from a naturally occurring signal peptide. In some aspects, the signal peptide is derived from a naturally occurring DP beta chain signal peptide. In some aspects, the signal peptide comprises a naturally occurring DP beta chain signal peptide. In some aspects, the signal peptide is derived from a naturally occurring DP alpha chain signal peptide. In some aspects, the signal peptide comprises a naturally occurring DP alpha chain signal peptide. In some aspects, the signal peptide is derived from a fibulin light chain (FibL) signal peptide. In some aspects, the signal peptide comprises SEQ ID NO: 9. In some aspects, the signal peptide is synthetic.

[0153] II.A.4. Transmembrane Domain

[0154] In some aspects, the DP β chain and / or DP α chain further comprises a transmembrane domain. The transmembrane domain can be of any length and of any origin. In some aspects, the transmembrane domain is at least about 1 to at least about 50 amino acids in length. 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 DP β chain transmembrane domain. In some aspects, the transmembrane domain comprises a naturally occurring DP β chain transmembrane domain. In some aspects, the transmembrane domain is derived from a naturally occurring DP α chain transmembrane domain. In some aspects, the transmembrane domain comprises a naturally occurring DP α chain transmembrane domain.

[0155] II.A.5. Leucine Zippers

[0156] In some aspects, the DP β chain and / or DP α chain further comprises one or more leucine zipper (LZip) sequences. Any LZip sequence known in the art can be used in the compositions and methods disclosed herein. In some aspects, the DP β chain and / or DP α chain comprises an acidic LZip (aLZip), a basic LZip (bLZip), or both. In some aspects, the one or more LZip sequences are derived from a naturally occurring LZip sequence. In some aspects, the one or more LZip sequences comprise a naturally occurring LZip sequence. In some aspects, the one or more LZip sequences are synthetic. In certain aspects, the one or more LZip sequences comprise the LZip sequence set forth in SEQ ID NO: 4 (Table 1).

[0157] II.A.6. Linkers

[0158] In some aspects, the DP β chain and / or DP α chain useful in the present disclosure further comprises a linker. Any linker known in the art can be used in the compositions and methods disclosed herein. In certain aspects, the linker comprises a Gly / Ser linker. In some aspects, the linker comprises an amino acid sequence selected from the group consisting of GlySer, Gly2Ser, Gly3Ser, and Gly4Ser. In some aspects, the linker is located N-terminal to the extracellular domain of the DP α chain or DP β chain. In some aspects, the linker is located C-terminal to the extracellular domain of the DP α chain or DP β chain. In some aspects, the linker is located between the extracellular domain and the transmembrane domain of the DP α chain or DP β chain. In some aspects, the linker is located between the extracellular domain and the one or more LZip sequences of the DP α chain or DP β chain. In some aspects, the linker is located between the extracellular domain and the signal peptide of the DP α chain or DP β chain.

[0159] 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 aspects, 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.

[0160] 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, 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.

[0161] II.B. Cells

[0162] In certain aspects of the disclosure, the MHC class II molecule of the disclosure is linked or associated with the membrane of a cell. In certain aspects, the beta chain of the MHC class II molecule is linked or associated with the membrane of a cell. In certain aspects, the alpha chain of the MHC class II molecule is linked or associated with the membrane of a cell. In certain aspects, the alpha chain and the beta chain of the MHC class II molecule are linked or associated with the membrane of a cell.

[0163] Certain aspects of the present disclosure relate to cells comprising the MHC class II molecules disclosed herein. Any cell can be used in the compositions described herein. In certain aspects, the cell is a mammalian cell. In some aspects, the cell is an insect cell. In some aspects, the cell is derived from a healthy cell, e.g., a healthy fibroblast. In some aspects, the cell is derived from a tumor cell. Non-limiting examples of cells that can be used in the present 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 certain aspects, the cell is any of the cells disclosed in Hasan et al., Adv. Genet. Eng. 4(3): 130 (2015), incorporated by reference in its entirety herein.

[0164] In certain aspects, the cell is a professional APC. In certain aspects, the cell is a macrophage, a B cell, a dendritic cell, or any combination thereof.

[0165] In certain aspects, the cell lacks endogenous expression of one or more MHC class II alleles. In some aspects, the cell lacks endogenous expression of an HLA-DP allele. In some aspects, the cell lacks endogenous expression of an HLA-DP a chain allele. In some aspects, the cell lacks endogenous expression of an HLA-DP b chain allele.

[0166] II.C. Soluble MHC class II molecules

[0167] In certain aspects, the MHC class II molecule is not associated with a cell membrane, e.g., the MHC class II molecule is in a soluble form. As used herein, a soluble MHC class II molecule includes any MHC class II molecule or portion thereof described herein that is not associated with a cell membrane. In certain aspects, the MHC class II molecule or portion thereof is not bound to any membrane. In some aspects, the MHC class II molecule or portion thereof is bound to an inert particle. In some aspects, the MHC class II molecule or portion thereof is bound to the membrane of an extracellular vesicle. In some aspects, the MHC class II molecule is bound to an artificial membrane or artificial surface, e.g., the surface of an array plate.

[0168] Any inert particle known in the art can be used in the compositions and methods of the present disclosure. In some aspects, the inert particle is a bead. In some aspects, the bead is a glass bead, a latex bead, a metal bead, or any combination thereof. In some aspects, the inert particle is a nanoparticle (NP). Any NP known in the art can be used in the compositions and methods of the present disclosure. In certain aspects, the nanoparticle is selected from the group consisting of polyethylene glycolated iron oxide, chitosan, dextran, gelatin, alginate, liposome, starch, branched polymer, carbon-based carriers, polylactic acid, poly(cyano)acrylate, polyethylenimine, block copolymer, polycaprolactone, SPIONS, USPIONS, Cd / Zn-selenide, or silica nanoparticles. In particular aspects, the nanoparticle is a polyethylene glycolated iron oxide nanoparticle. Non-limiting examples of nanoparticles that can 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 by reference herein in their entirety.

[0169] In some aspects, the MHC class II molecule comprises a fragment of a full-length MHC class II molecule, wherein one or more amino acids of the transmembrane domain of the alpha chain and / or the transmembrane domain of the beta chain are deleted. In some aspects, the MHC class II molecule comprises an extracellular domain of the alpha chain (e.g., as set forth in SEQ ID NO: 6) and / or an extracellular domain of the beta chain (e.g., as set forth in SEQ ID NO: 1 or 3). In certain aspects, the MHC class II molecule comprises a DP alpha 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 to SEQ ID NO: 6. In some aspects, the MHC class II molecule comprises a DP alpha chain comprising the amino acid sequence set forth in SEQ ID NO: 6.

[0170] In certain aspects, the MHC class II molecule comprises a DP beta 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 to SEQ ID NO: 1. In some aspects, the MHC class II molecule comprises a DP beta chain comprising the amino acid sequence set forth in SEQ ID NO: 1. In certain aspects, the MHC class II molecule comprises a DP beta 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 to SEQ ID NO: 3. In some aspects, the MHC class II molecule comprises a DP beta chain comprising the amino acid sequence set forth in SEQ ID NO: 3. In certain aspects, the MHC class II molecule comprises a DP beta 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 to SEQ ID NO: 4. In some aspects, the MHC class II molecule comprises a DP beta chain comprising the amino acid sequence set forth in SEQ ID NO: 4. In certain aspects, the MHC class II molecule comprises a DP beta 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 to SEQ ID NO: 5. In some aspects, the MHC class II molecule comprises a DP beta chain comprising the amino acid sequence set forth in SEQ ID NO: 5.

[0171] II.D. NUCLEIC ACID MOLECULES AND VECTORS

[0172] Certain aspects of the present disclosure relate to nucleic acid molecules encoding the MHC class II molecules disclosed herein. In some aspects, the nucleic acid molecule encodes a MHC class II beta chain disclosed herein. In certain aspects, the nucleic acid molecule encoding the MHC class II beta 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 to the sequence set forth in SEQ ID NO: 2.

[0173] In some aspects, the nucleic acid molecule encodes a MHC class II alpha chain disclosed herein. In certain aspects, the nucleic acid molecule encoding the MHC class II alpha 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 to the sequence set forth in SEQ ID NO: 7.

[0174] In some aspects, the nucleic acid molecule encodes both a MHC class II alpha chain disclosed herein and a MHC class II beta chain disclosed herein. In some aspects, the sequence encoding the MHC class II alpha chain is under the control of the same promoter as the sequence encoding the MHC class II beta chain. In some aspects, the sequence encoding the MHC class II alpha chain is under the control of a first promoter and the sequence encoding the MHC class II beta chain is under the control of a second promoter.

[0175] In some aspects, the disclosure relates to a first nucleic acid molecule encoding a MHC class II beta chain disclosed herein and a second nucleic acid molecule encoding a MHC class II alpha chain disclosed herein.

[0176] Certain aspects of the disclosure relate to a vector or a set of vectors comprising a nucleic acid molecule 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.

[0177] In certain aspects, the vector is a retroviral vector. In some aspects, the vector is an adenoviral vector, a lentivirus, a Sendai virus, a baculovirus vector, an Epstein Barr virus vector, a papovaviral vector, a vaccinia virus vector, a herpes simplex virus vector, or an adeno-associated virus (AAV) vector. In particular aspects, the vector is an AAV vector. In some aspects, the vector is a lentivirus. In particular aspects, the vector is an adenoviral vector. In some aspects, the vector is a Sendai virus. In some aspects, the vector is a hybrid vector. Examples of hybrid vectors that can be used in the present disclosure can be found in Huang and Kamihira, Biotechnol. Adv. 31(2):208-23 (2103), which is incorporated by reference herein in its entirety.

[0178] III. Methods of the disclosure

[0179] Certain aspects of the disclosure relate to methods of treating a disease or a condition in a subject. In some aspects, the disclosure relates to methods of enhancing an immune response in a subject in need thereof.

[0180] III. A. Methods of treating a tumor

[0181] Certain aspects of the present disclosure relate to methods of treating a cancer in a subject in need thereof, 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.

[0182] In some aspects, the cancer is selected from melanoma, bone cancer, kidney cancer, prostate cancer, breast cancer, colon cancer, lung cancer, cutaneous or intraocular malignant melanoma, pancreatic cancer, skin cancer, cancer of the head or neck, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, 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), cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, acute or chronic leukemia, acute myelogenous leukemia (AML), chronic myelogenous leukemia, acute lymphoblastic leukemia (ALL) (including non T-cell ALL), chronic lymphocytic leukemia (CLL), solid tumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumors, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, other B-cell malignancies, and any combination of these cancers. In some aspects, the cancer is melanoma.

[0183] In some aspects, the cancer is recurrent. In some aspects, the cancer is refractory. In some aspects, the cancer is advanced. In some aspects, the cancer is metastatic.

[0184] In some aspects, the methods disclosed herein treat a cancer in a subject. In some aspects, the methods disclosed herein reduce the severity of one or more symptoms of a cancer. In some aspects, the methods disclosed herein reduce the size or number of tumors derived from the cancer. In some aspects, the methods disclosed herein increase the overall survival of a subject relative to a subject not provided the methods disclosed herein. In some aspects, the methods disclosed herein increase the progression-free survival of a subject relative to a subject not provided the methods disclosed herein. In some aspects, the methods disclosed herein result in a partial response in a subject. In some aspects, the methods disclosed herein result in a complete response in a subject.

[0185] Certain aspects of the present disclosure relate to methods of treating an infection in a subject in need thereof, 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. Non-limiting examples of infections that can be treated using the compositions and methods disclosed herein include infections of viruses (including viroids and prions), bacteria, fungi, parasites, or any combination thereof. In some aspects, the virus is a herpes virus, HIV, a papovavirus, a morbillivirus, a rubivirus, a human papillomavirus (HPV), human T-lymphotrophic virus 1, Epstein-Barr virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, influenza virus, norovirus, and any combination thereof. In some aspects, the bacteria is selected from the group consisting of Streptococcus, Staphylococcus, and Escherichia coli. In some aspects, the bacterial infection is selected from the group consisting of brucellosis, Campylobacter infection, cat scratch disease, cholera, E. coli infection, gonorrhea, Klebsiella infection, Enterobacter infection, Serratia infection, Legionella infection, meningococcal infection, pertussis, plague, Pseudomonas infection, Salmonella infection, Shigellosis, typhoid fever, tularemia, anthrax, diphtheria, Enterococcus infection, Erysipelothricosis, Listeriosis, Nocardiosis, Pneumococcal infection, Staphylococcus infection, Streptococcus infection, and any combination thereof. In some embodiments, the parasitic infection is selected from the group consisting of enterobiasis, trichomoniasis, toxoplasmosis, giardiasis, cryptosporidiosis, malaria, hookworm disease, ringworm, tapeworm disease, fluke disease, and any combination thereof. In some aspects, the fungal infection is selected from the group consisting of Candida, Malassezia furfur, dermatophytes (e.g., Epidermophyton, Microsporum, and Trichophyton), or any combination thereof.

[0186] In some aspects, the methods disclosed herein comprise treating a cancer or an infection in a subject in need thereof, the methods comprising administering to the subject a cell described herein, wherein the cell comprises an MHC class II molecule disclosed herein, a nucleic acid molecule disclosed herein, a vector disclosed herein, or any combination thereof.

[0187] In some aspects, the cell is obtained from the subject. In some aspects, the cell is obtained from a donor other than the subject.

[0188] III.B. Methods of enriching a target T cell population

[0189] Certain aspects of the present disclosure relate to methods of enriching a target T cell population obtained from a human subject. In some aspects, the methods comprise contacting the T cells with an HLA class II molecule disclosed herein. In some aspects, the methods comprise contacting the T cells with a cell (e.g., an APC) disclosed herein. In some aspects, following the contacting, the enriched T cell population comprises a greater number of T cells capable of binding to the HLA class II molecule relative to the number of T cells capable of binding to the HLA class II molecule prior to the contacting.

[0190] Some aspects of the disclosure relate to methods of selecting T cells that are capable of targeting diseased cells, e.g., tumor cells. In some aspects, the methods comprise contacting a population of isolated T cells in vitro with a complex comprising an MHC class II molecule disclosed herein and a fragment of a polypeptide, e.g., an antigen expressed by a diseased cell, e.g., a polypeptide expressed by a tumor, e.g., an epitope. In some aspects, the T cells are obtained from a human subject.

[0191] The T cells obtained from a human subject can be any T cell disclosed herein. In some aspects, the T cells obtained from a human subject are tumor infiltrating lymphocytes (TILs).

[0192] In some aspects, the methods further comprise administering the enriched T cells to a human subject. In some aspects, the subject is preconditioned prior to receiving the T cells as described herein.

[0193] All of the various aspects, aspects, and options described herein can be combined in any and all variations.

[0194] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0195] Having generally described this disclosure, a further understanding can be obtained by reference to the examples provided herein. These examples are for illustrative purposes only and are not intended to limit the disclosure.

[0196] Examples

[0197] Example 1 - Methods

[0198] Cells

[0199] Peripheral mononuclear cells were obtained by density gradient centrifugation (Ficoll-Paque PLUS, GE Healthcare Life Sciences, Marlborough, MA). The K562 cell line is a red-white blood cell line with defective HLA class I / II expression. K562-based artificial APCs (aAPCs) expressing various HLA class II genes as single HLA alleles along with CD80 and CD83 have been previously reported (Butler et al., PloS One 7, e30229 (2012)). The Jurkat 76 cell line is a T cell leukemia cell line that lacks endogenous TCR, CD4, and CD8 expression. Jurkat 76 / CD4 cells were generated by retroviral transduction of the human CD4 gene. A375, SK-MEL-21, SK-MEL-28, SK-MEL-37, and Me275 are melanoma cell lines. HEK293T cells and melanoma cell lines were grown in DMEM supplemented with 10% FBS and 50 pg / ml gentamicin (ThermoFisher Scientific, Waltham, MA). K562 and Jurkat 76 cell lines were cultured in RPMI 1640 supplemented with 10% FBS and 50 pg / ml gentamicin.

[0200] Peptides

[0201] Synthetic peptides were purchased from Genscript (Piscataway, NJ) and dissolved in DMSO at 50 pg / ml. Peptide sequences are shown in Table 3.

[0202] Table 3: Synthetic peptide sequences

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209] Genes

[0210] Novel TCR genes were cloned using the SMARTer RACE 5' / 3' kit (Takara Bio, Shiga, Japan) via 5'-rapid amplification (RACE) PCR of the cDNA ends and sequenced as previously described. All genes were cloned into the pMX retroviral vector and transduced into cell lines using a retroviral system based on 293GPG and PG13 cells.

[0211] Antibody

[0212] The following antibodies were used for flow cytometry analysis: PE-conjugated anti-class II (9-49(I3)) and APC-Cy7-conjugated anti-CD4 (RPA-T4, Biolegend, San Diego, CA). 44 FITC-conjugated anti-NGFR (ME20.4, Biolegend, San Diego, CA), PE-conjugated anti-His tag (AD1.1.10, Abcam, Cambridge, MA), and FITC-conjugated anti-Vβ22 (IMMU 546, Beckman Coulter, Brea, CA). According to the manufacturer's instructions, PE-conjugated streptavidin (Thermo Fisher Scientific, Waltham, MA) was used to bind biotinylated DP4 / NY-ESO1. 157-170 and DP4 / WT1 329-348单 Cell multiplication was performed. 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).

[0213] The following antibodies were used for immunoblot analysis: anti-β-actin (C4, Santa Cruz Biotechnology, Santa Cruz, CA), rabbit polyclonal anti-MAGE-A2 (Abeam, Cambridge, MA), anti-CCND1 (EPR2241, Abeam, Cambridge, MA), HRP-conjugated goat anti-mouse IgG (H+L) secondary antibody (Promega, Fitchburg, WI), and HRP-conjugated anti-rabbit IgG (H+L) secondary antibody (Promega, Fitchburg, WI).

[0214] TCR transduction into primary T cells

[0215] Pan T cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) and CD4 + T cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) were used to purify CD3 + and CD4 + T cells. Purified T cells were stimulated with 200 Gy irradiated aAPC / mOKT3 at an E:T ratio of 20: 1. From the next day, activated T cells were retrovirally transduced with cloned TCR genes by centrifugation at 1,000 x g for 1 hour at 32 °C for 3 consecutive days or using Retronectin-coated plates (Takara Bio, Shiga, Japan). On the second day, 100 IU / ml IL-2 and 10 ng / ml IL-15 were added to TCR transduced T cells. Medium was replenished every 2-3 days.

[0216] Staining with soluble CD4

[0217] The soluble CD4 (sCD4) gene was generated by fusing the human CD4 extracellular domain with a 6xHis tag via a GS linker. HEK293T cells were retrovirally transduced with the sCD4 gene and culture supernatant containing sCD4 monomers was harvested. sCD4 was dimerized with a PE-labeled anti-6xHis tag mAb (AD1.1.10, Abeam, Cambridge, MA) and used. K562 cells expressing HLA class II were stained with dimerized sCD4 in the presence of goat serum for 30 minutes at room temperature. Surface HLA class II expression in K562-derived cells expressing various class II genes was determined by flow cytometry as shown in Figures 13A-13Q .

[0218] Construction and screening of a multi-site directed DPB1*04:01 mutant cDNA library

[0219] Multi-site directed random mutagenesis was inserted into DPB1*04:01 cDNA by using PCR and the following primer sets: for L112 and V114, forward: 5'-CACCACAACNNNCTTNNNTGCCACGTG-3' (SEQ ID NO: 12) and reverse: 5'-CACGTGGCANNNAAGNNNGTTGTGGTG-3' (SEQ ID NO: 13); for V141, forward: 5'-ACAGCTGGGGTCNNNTCCACCAACCTG-3' (SEQ ID NO: 14) and reverse: 5'-CAGGTTGGTGGANNNGACCCCAGCTGT-3' (SEQ ID NO: 15); for L156 and M158, forward: 5'-CAGATCNNNGTGNNNCTGGAAATGACC-3' (SEQ ID NO: 16) and reverse: 5'-GGTCATTTCCAGNNNCACNNNGATCTG-3' (SEQ ID NO: 17). N represents any nucleotide. The resulting PCR fragments were fused to each other to construct a mutant full-length DPB1*04:01 cDNA expression library, which carries random mutations at positions L112, V114, V141, L156, and M158. K562 cells stably expressing DPA1*01:03 genes were infected with recombinant retroviruses produced by packaging cell line 293GPG at a transduction efficiency of less than 30%. The infected K562 cells were stained with soluble CD4 dimers, and dimer-positive cells were collected using a flow cytometry cell sorter. Mutant DPB1*04:01 genes were cloned from the collected cells and transduced into K562 cells by retrovirus together with the wild-type DPA1*01:03 gene described above.

[0220] Generation of HLA class II monomers and dimers

[0221] The extracellular domain of the wild-type class II alpha gene was fused via a GGGS linker to an acidic leucine zipper, followed by a GS linker to a 6xHis tag (see SEQ ID NO: 8). The extracellular domain of the class II beta gene carrying the mutation (see SEQ ID NO: 3) was similarly linked via a GGGS linker (see SEQ ID NO: 4) to a basic leucine zipper. HEK293T cells were transfected with the alpha and beta genes using a retroviral system based on 293GPG cells and cultured in DMEM supplemented with 10% FBS and 50 μg / ml gentamicin. For DP4 dimer staining, K562 cells stably expressing DPA1*01:03 genes were infected with recombinant retroviruses produced by packaging cell line 293GPG at a transduction efficiency of less than 30%. The infected K562 cells were stained with soluble DP4 dimers, and dimer-positive cells were collected using a flow cytometry cell sorter. Mutant DPB1*04:01 genes were cloned from the collected cells and transduced into K562 cells by retrovirus together with the wild-type DPA1*01:03 gene described above. L112W / V141MHEK293T cells were grown to confluency and the media was replaced with serum-free 293 SFM II media (Thermo Fisher Scientific, Waltham, MA). After 48 hours, the conditioned media was harvested and concentrated using Amicon Ultrafilters (10 kDa molecular weight cut-off (MWCO)) (MilliporeSigma, Burlington, MA). The supernatant containing soluble HLA class II was then mixed with 100 pg / ml of the peptide of interest at 37 °C for 20-24 hours to perform in vitro peptide exchange. Monomers that did not undergo peptide exchange were used as a control. The concentration of monomers was measured by specific ELISA using nickel-coated plates (XPressBio, Frederick, MD) and anti-His tag biotinylated mAb (AD1.1.10, R&D Systems, Minneapolis, MN). Soluble HLA class II monomers were dimerized at a 2: 1 molar ratio using PE-conjugated anti-His mAb (AD1.1.10, Abeam, Cambridge, MA) at 4 °C for 1.5 hours to perform staining.

[0222] DP4-restricted antigen-specific CD4 + Stimulation of T cells

[0223] Using CD4 + T cell isolation kit (Miltenyi Biotec, Bergisch Gladbach, Germany) to purify CD4 + T cells. Purified T cells were stimulated with DP4-expressing aAPCs pulsed with DP4-restricted peptide at 10 pg / ml and irradiated at 200 Gy at an E:T ratio of 20: 1. Forty-eight hours later, 10 IU / ml IL-2 and 10 ng / ml IL-15 were added to the CD4 + T cells. Medium supplemented with IL-2 (10 IU / ml) and IL-15 (10 ng / ml) was replenished every 2-3 days. After 2 weeks of stimulation, T cells were stained for DP4 L112W / V141M dimer staining.

[0224] HLA class II dimer and tetramer staining

[0225] Primary T cells and Jurkat 76 / CD4 T cells transduced with exogenous TCR genes were incubated with 10 pg / ml of the peptide of interest at 37 °C for 20-24 hours to perform in vitro peptide exchange. Monomers that did not undergo peptide exchange were used as a control. The concentration of monomers was measured by specific ELISA using nickel-coated plates (XPressBio, Frederick, MD) and anti-His tag biotinylated mAb (AD1.1.10, R&D Systems, Minneapolis, MN). Soluble HLA class II monomers were dimerized at a 2: 1 molar ratio using PE-conjugated anti-His mAb (AD1.1.10, Abeam, Cambridge, MA) at 4 °C for 1.5 hours to perform staining. 46Cells were pre-treated with 50 nM dasatinib (LC Laboratories, Woburn, MA) for 30 min and stained with 5-15 pg / ml class II dimer at room temperature for 4-5 h. After washing, cell surface molecules were re-stained with APC-Cy7-conjugated anti-CD4 mAb, FITC-conjugated anti-NGFR mAb, and PE-conjugated anti-Vp22 mAb.

[0226] ELISPOT assay

[0227] Cytokine ELISPOT assays were performed as previously reported (see, e.g., Yamashita et al., Nat. Commun. 8: 15244 (2017); and Anczurowski et al., Sci. Rep. 8: 4804 (2018)).

[0228] Immunoblotting

[0229] Immunoblotting analysis was performed as previously reported (see, e.g., Yamashita et al., Nat. Commun. 8: 15244 (2017); and Anczurowski et al., Sci. Rep. 8: 4804 (2018)).

[0230] Protein modeling

[0231] HLA-DP4 and human CD4 complex model structures were predicted based on the structures of PDB ID 3S5L and 3T0E using Swiss-Model workspace for quaternary structure prediction.

[0232] Statistical analysis

[0233] Statistical analysis was performed using GraphPad Prism 6.0 software (GraphPad Software, San Diego, CA). Two-sample comparisons were performed using unpaired, two-tailed Student’s t test. No statistical methods were used to predetermine sample size. The investigator was not blinded to the allocation during experiments or outcome assessment. Experiments were not randomized.

[0234] Biolayer interferometry sensorgrams

[0235] The extracellular domain of human CD4 (residues 26-440 of NP_000607.1) followed by a GS linker and a 10x histidine (His) tag was stably expressed in the human cell line A375 (SEQ ID NOs:245-246; Table 4). Recombinant 10x His-tagged CD4 protein was purified from the supernatant using TALON metal affinity resin (Takara Bio, Shiga, Japan). Eluted protein was concentrated using Amicon Ultra-15 centrifugal columns with 10 kDa MWCO (MilliporeSigma, Burlington, MA). Buffer exchange was performed to HBS-EP (GE Healthcare Life Sciences, Marlborough, MA) using 10 kDa MWCO mini-dialyzers (Thermo Fisher Scientific, Waltham, MA). Purity of the recombinant CD4 protein was consistently >90% as confirmed by SDS-PAGE.

[0236] Recombinant DP4 protein was composed of the extracellular domain of DPA1*01:03 and either wild-type DPB1*04:01 or the L112W / V141M mutant. DPA1*01:03 was followed by an acidic leucine zipper, a GS linker, and a 10x histidine tag, while wild-type and mutant DPB1 were followed by a basic leucine zipper, a GS linker, and a biotinylation sequence (GLNDIFEAQKIEWHE; SEQ ID NO:244). Both DPA and DPB genes were stably expressed in A375-BirA cells, which were transduced with a codon-optimized BirA gene encoding a 5’-terminal leader sequence and a 3’-terminal ER retention KDEL motif. Recombinant DP4 protein was purified from the supernatant using TALON metal affinity resin (Takara Bio, Shiga, Japan). Eluted protein was concentrated using Vivaspin 500 centrifugal columns with 10 kDa MWCO (GE Healthcare Life Sciences, Marlborough, MA) and resuspended to working volume in PBS.

[0237] Wild-type DP4 and DP4 L112W / V141MBinding to CD4. Experiments were performed at 25 °C using 96-well OptiPlates (Perkin Elmer, Waltham, MA) with a sample volume of 200 μΐ and constant shaking at 1,000 rpm. Biotinylated recombinant DP4 was loaded onto a streptavidin-coated biosensor (ForteBio, Fremont, CA) until saturation, followed by baseline measurements in HBS-EP buffer. Association was measured by incubating the loaded sensor with titrated concentrations of recombinant CD4 (0.8125 to 26 μΜ) for 400 s, followed by dissociation in separate HBS-EP buffer for 300 s. Steady-state analysis was fitted using the one-site specific binding model in GraphPad Prism 7.0

[0238] Table 4. Soluble 10x His-tagged CD4 nucleic acid sequence

[0239]

[0240]

[0241] Example 2 - L112W / V141M substitutions in the DP β chain enhance DP binding to CD4

[0242] A cDNA expression library of DPB1*04:01 (DP4 β) genes carrying random mutations at L112, V114, V141, L156, and M158 (corresponding to L114, V116, V143, L158, and M160 of the DR1 β chain, respectively) was generated and co-expressed with a wild-type DPA1*01:03 (DP α) gene in class II-deficient K562 cells. After two rounds of screening using a soluble CD4 protein (sCD4), a cell population with enhanced CD4 binding was isolated, from which a mutant DP4 β gene carrying L112W, V114M, V141M, and M158I substitutions was molecularly cloned. When ectopically expressed in K562 cells, a mutant DP4 molecule composed of a wild-type DP α chain and the cloned mutant DP4 β chain carrying L112W, V114M, V141M, and M158I substitutions (DP4 L112W / V114M / V141M / M158I ) indeed showed enhanced binding to sCD4 compared to a wild-type DP4 molecule, thus ruling out the possibility that the enhanced CD4 binding was an artifact of the screening process Figures 1A-1F

[0243] ​To determine which of the four mutations was crucial for enhanced CD4 binding, reversion mutagenesis was performed. All possible reversion DP4 mutants were reconstructed in class II negative K562 cells and stained with sCD4. Both L112W and V141M, rather than single substitutions of V114M or M158I, enhanced DP4 binding to sCD4. Figure 1G Importantly, the L112W / V141M double mutation (DP4) L112W / V141M ) Synergistically enhanced DP4 / CD4 binding ( Figure 1G Interestingly, both the V114M and M158I single-replacement displays support DP4. L112W / V141M Enhanced binding achieved through mutation has negative effects. Figure 1G Previous studies estimated the K-lineage between CD4 and HLA class II. D Value >2mM. DPR was measured using biolayer interferometry (BLI) combined with assay. L112 / V141M Affinity to CD4. Although no binding was detected between wild-type DP4 and CD4, DP4... L112W / V141M With K at 8.9 μM ± 1.1 D Combined with CD4 ( Figure 1H and Figures 1X-1BK This value indicates that binding affinity has increased by at least 200-fold. Furthermore, CD4 and DP4 were observed... L112W / V141M The affinity between DP4 and HLA class I is higher than that between human CD8 and HLA class I (approximately 200 μM), and comparable to that between mouse CD8 and mouse MHC class I (approximately 10 μM). To confirm DP4... L112W / V141M Enhanced binding between CD4 and CD4 results in enhanced CD4. + T-cell response, using CD4 transduced with DP4 / WT1 TCR (clone 9). - and CD4 + Jurkat 76T cells act as responders to wild-type DP4 or DP4 expressing a single class II allele. L112W / V141M The immunostimulatory capacity of artificial APCs (aAPCs) was compared. As expected, those carrying DP4... L112W / V141M aAPCs exhibit enhanced T cell stimulation activity in a CD4-dependent manner. Figure 1I ).

[0244] Next, other DP alleles of CD4 were analyzed to determine whether the L112W / V141M mutation also enhanced binding. Although wild-type DP2, DP5, or DP8 do not bind to CD4, all three molecules showed strong binding to CD4 when the L112W / V141M double mutation was introduced into the DPβ chain of these molecules. Figures 1J-1W ). A structural model built based on previous reports (Figures 2A-2D ) reveals that in DP4 L112W / V141M In the CD4 complex, the two L112W / V141M mutations significantly induced hydrophobic effects at the K35, Q40, and T45 positions of CD4. These results indicate that the L112W / V141M mutation can enhance CD4 binding of at least all four tested DP alleles.

[0245] Example 3 – Affinity-matured DP4 L112W / V141M Multimer-specific staining homologous TCR

[0246] To determine the effect of the L112W / V141M double mutation of DP4β on DP4 multimer staining, soluble DP4 was produced. L112W / V141M单 The cells were then dimerized with an anti-His tag mAb. Primary T cells were transduced with three different DP4-restricted TCRs specific to MAGE-A3 (clone R12C9), WT1 (clone 9), and NY-ESO-1 (clone 5B8), respectively, and then homologous DP4 was used. L112W / V141M二 Polymer staining. As shown in Figures 3A to 3P, each DP4... L112W / V141M二 Polymer-specific staining of CD4 expressing homologous TCR + T cells. Anti-Vβ22 mAb and anti-NGFR mAb were administered along with their respective DP4 groups. L112W / V141M二 Complex staining of R12C9 and clone 9 transduced T cells with polymerase chain reaction (PCR) confirmed that almost all TCR-transduced CD4+ cells were transduced. + T cells were successfully used with the corresponding DP4 L112W / V141M二 Polymer staining (Figs. 4A to 4H). Our novel DP4 tetramer, compared to the conventional wild-type DP4 tetramer, shows... L112W / V141M The dimer showed superior staining performance on both DP4 / WT1 and DP4 / NY-ESO-1 T cells compared to conventional wild-type DP4 tetramer (Figs. 5A to 5P). Notably, even at the highest available concentration, conventional wild-type DP4 / NY-ESO-1 tetramer failed to stain homologous T cells (data not shown).

[0247] Example 4 – DP4 L112W / V141M Dimer technology is robust and versatile.

[0248] To prove DP4 L112W / V141M The robustness and versatility of multimer staining were demonstrated, and a comprehensive screening of the in vitro immunogenicity of potential DP4-restricted peptides derived from a range of tumor-associated antigens was conducted (Table 3). 196 DP4-restricted and tumor-associated antigen-derived 20-mer peptides were predicted using a peptide prediction algorithm (NetMHC2 ver. 2.2) and then chemically synthesized (Table 3). Peripheral antigen-specific CD4+ T cells are generally low in frequency; therefore, primary T cells from 6 DP4 + melanoma patients were isolated + T cells were stimulated once with DP4-aAPC, pulsed with 196 peptides, respectively, and stained with cognate DP4 L112W / V141M dimers. To avoid potential in vitro priming, weak stimulation conditions were used. As Figures 6A-6F shown, 103 predicted DP4 peptides were at least in vitro immunogenic.

[0249] To validate the dimer staining results, we cloned seven DP4-restricted TCR genes 219-238 specific for CCND1 225-244 , HSD17B12 296-315 , LGSN 108-127 , MAGE-A2 4922-4941 and MUC5AC + from dimer-positive T cells (Table 5 and Table 5). When cloned and reconstituted in human CD4 L112W / V141M TCR-deficient T cells, all these TCRs were successfully stained by cognate DP4 219-238、06 dimers (Figures 8A-8X) and functioned in a DP4-restricted and antigen-specific manner Figures 7A-7L ). Figures 9A-9G

[0250] Among the four TCRs expressed in primary T cells, respectively, three, 03-CCND1 108-127 , 04-HSD17B12 4922-4941 and 05-MUC5AC 108-127 , were able to recognize cognate peptides endogenously processed and presented by DP4 (Figures 10A-10C, 11A-11C and 12A-12C, respectively). Importantly, 06-MAGE-A2 L112W / V141M transduced primary T cells were able to recognize melanoma cell lines in a DP4 and MAGE-A2 dependent manner Figure 11A . Figures 12A-12E

[0251] Table 5: DP4-restricted TCRs

[0252]

[0253] The role and function of CD4 as a co-receptor compared to CD8 are not fully elucidated. This lack of information is mainly due to the exceptionally weak binding between CD4 and class II, which greatly limits the study of the binding interaction between CD4 and class II. In this study, an affinity-matured form of HLA-DP4, DP4 L112W / V141M , was isolated by enhanced CD4 binding and a novel DP4​​L112W / V141M dimer technology that introduced detection of DP4-restricted antigen-specific CD4 + robustness and stringency of T cells.

[0254] Using this DP4 L112W / V141M dimer technology, DP4-restricted anti-tumor T cell responses were comprehensively studied in vitro and multiple DP4-restricted immunogenic peptides and cognate TCR genes were identified. HLA-DP4 is the most prevalent HLA allele in many ethnic populations and belongs to the DP 84Gly group. Unlike other class II molecules, DP 84Gly molecules constitutively present peptides derived from endogenous sources regardless of invariant chain and HLA-DM expression. Improving endogenous peptide presentation by class II is associated with improved survival in cancer patients. Notably, the first human class II restricted TCR gene therapy did target a DP4-restricted MAGE-A3 peptide (see, e.g., Yao et al., J. Immunother. 39:191-201 (2016)). DP 84Gly genotypes such as DP2 and DP4 act as risk alleles for anti-neutrophil cytoplasmic autoantibody associated vasculitis. DP4 molecules can constitutively present peptides derived from endogenous tumor associated antigens and can induce more clinically relevant anti-tumor responses than other class II molecules, acting as a protective class II allele.

[0255] To identify affinity matured class II molecules, this example details multiple mutations in the beta chain but not the alpha chain, as the beta chain has more direct interaction with CD4 than the alpha chain. Additional mutations to the alpha and / or beta chain can further enhance binding between class II and CD4. However, using this soluble class II molecule with excessive CD4 binding capacity can result in non-specific staining of CD4 + T cells, thereby having a detrimental effect.

[0256] In summary, CD4 + T cells play a critical role in the development of autoimmune diseases as well as protection against pathogenic infections and cancer. The novel HLA class II multimer technology described herein can better facilitate the study of HLA class II restricted CD4 + T cell responses to HLA-DP alleles.

[0257] Example 5 - DP4 L112W / V141M specificity and binding

[0258] DP4 multimer staining of endogenous (untransduced) antigen-specific CD4 + T cells was analyzed. The novel DP4 L112W / V141MDimers for endogenous TRPC1 578-597 Specific CD4 + Positive staining of T cells ( Figures 14A-14B ) compared to the conventional DP4dextramer ( Figures 14C-14D Stronger than conventional tetramers ( ). Figures 15C-15D ) or dextramer ( Figures 15E-15F Compared to DP4 L112W / V141M The dimer showed significantly improved endogenous (untransduced) NY-ESO-1 157-170 Specific CD4 + T cell staining ( Figures 15A-15B (Table 6).

[0259] Table 6: DP4-Restricted TCR

[0260]

[0261]

[0262] Next, without in vitro stimulation, DP4, which is specific to a range of pathogen-related peptides, was used. L112W / V141M Dimer for memory CD4 + T cells were stained in vitro. Tetanus toxin was used for this purpose. 948-968 (TT 948-968 Herpes simplex virus type 2-UL21 283-302 (HSV-2-UL21 283-302 ) and respiratory syncytial virus glycoprotein 162-175 (RSV-GP 162-175 CD4 + A small subset of T cells was influenced by DP4. L112W / V141M Dimer positive staining (Figs. 16A to 16Y). Next, we used RSV-GP... 162-175 ( Figures 17A-17V ) and TT 948-968二 polymer + CD4 + Restrictive dilution of T cells to establish endogenous (untransduced) single-cell clones Figures 18A-18R These T cell clones showed that they produced IL-2 in an antigen-specific manner. Figure 17W and Figure 18S From DP4 L112W / V141M RSV-GP and TT dimer + Multiple TCRαβ pairs were isolated from single-cell clones, including one dominant pair (Table 6). Figures 17A-17W and Figures 18A-18S In the middle, via RSV-GP 162-175 and TT 948-968二Polymer + Limiting dilution of cells established single cell clones. When these RSV-GP and TT dimers + Single cell clones were stained with three different DP4 multimers (DP4 L112W / V141M DP4 L112W / V141M二 Polymer showed better staining of RSV-GP-(c12 and c39) and TT specific clones (c2 and c9) than conventional wild-type DP4 RSV-GP dextramer and wild-type DP4 TT tetramer and dextramer (Figures 19A-19NN).

[0263] Methods

[0264] Cells

[0265] Peripheral mononuclear cells were obtained by density gradient centrifugation. K562-based artificial antigen presenting cells (aAPCs) expressing various HLA class II genes as individual HLA alleles along with CD80 and CD83 have been previously reported (Butler, M. O., et al., PLoS One 7, e30229 (2012)). HEK293T cells were grown in DMEM supplemented with 10% FBS and 50 pg / ml gentamicin.

[0266] Peptides / antibodies

[0267] Synthetic peptides were dissolved in DMSO at 50 mg / ml. The following antibodies were used for flow cytometry analysis: APC-Cy7 conjugated anti-CD4 and PE conjugated anti-His tag.

[0268] Generation of HLA class II monomers and dimers

[0269] HEK293T cells were transfected with a and b genes using a 293GPG cell-based retroviral system (see Hirano, N., et al., Blood 107, 1528-1536 (2006); Butler, M. O., et al., Clin Cancer Res 13, 1857-1867 (2007); Hirano, N., et al., Blood 108, 2662-2668 (2006)) and cultured in DMEM supplemented with 10% FBS and 50 pg / ml gentamicin. For DP4 dimer staining, HEK293T cells stably secreting soluble DP4 L112W / V141MHEK293T cells were grown to confluency and the media was replaced with serum-free 293SFM II media (Thermo Fisher Scientific, Waltham, MA). After 48 hours, the conditioned media was harvested and concentrated using Amicon Ultrafilters (10 kDa molecular weight cut-off (MWCO)) (MilliporeSigma, Burlington, MA). The supernatant containing soluble HLA class II was then mixed with 100 pg / ml of the peptide of interest at 37°C for 20-24 hours to perform in vitro peptide exchange. Monomer concentration was measured by specific ELISA using nickel-coated plates and anti-His tag biotinylated mAb. Soluble HLA class II monomers were dimerized using PE-conjugated anti-His mAb at a 2: 1 molar ratio for 1.5 hours at 4°C for staining.

[0270] DP4-restricted antigen-specific CD4 + Stimulation of T cells

[0271] CD4 + T cells were purified and stimulated with DP4-expressing aAPC pulsed with DP4-restricted peptides at 10 pg / ml and irradiated at 200 Gy at an E:T ratio of 20: 1. After 48 hours, 10 IU / ml IL-2 and 10 ng / ml IL-15 were added to the CD4 + T cells. Medium supplemented with IL-2 (10 IU / ml) and IL-15 (10 ng / ml) was replenished every 2-3 days. After 2 weeks of stimulation, T cells were stained for DP4 L112W / V141M dimer.

[0272] HLA class II dimer, tetramer, and dextramer staining

[0273] DP4 tetramer and dextramer were compared in multimer staining analysis. Primary CD4 + T cells transduced with antigen-specific TCR genes were pre-treated with 50 nM dasatinib for 30 minutes at 37°C and stained with 5-15 pg / ml class II dimers for 4-5 hours at room temperature. After washing, cells were counterstained for cell surface molecules with APC-Cy7-conjugated anti-CD4 mAb.

[0274] Unstimulated CD4 + T cells from melanoma patients

[0275] One million CD4 +T cells were purified and pre-treated with 50 nM dasatinib for 30 min at 37°C. Cells were stained with 5-15 pg / ml class II dimers for 4-5 h at room temperature. After washing, cells were re-stained for cell surface molecules with APC-Cy7-conjugated anti-CD4 mAb. Dimers + Absolute counts of cells.

[0276] DP4 dimers + Expansion of T cells and establishment of single T cell clones.

[0277] To expand DP4 L112W / V141M dimers + T cells, as described above, were CD4 + T cells were stimulated and expanded with DP4 L112W / V141M dimers. Dimers + Cells were sorted by using anti-PE magnetic beads and expanded by using artificial APCs / mOKT3 irradiated at 200 Gy with an E:T ratio of 5-20:1 (see Butler, M.O. et al. PLoS One 7, e30229 (2012)). Medium was replenished every 2-3 days with IL-2 (10 IU / ml) and IL-15 (10 ng / ml). After two to three weeks, T cells were stained with DP4 L112W / V141M dimers. DP4 L112W / V141M dimers + single cell clones were generated by limiting dilution as previously described (see Su, L.F. et al. Immunity 38, 373-383 (2013)). Briefly, DP4 L112W / V141M dimers were used to purify and stain memory CD4 + T cells without dasatinib pre-treatment. Dimers + Cells were sorted and then stimulated in 96-well plates with 5 pg / ml PHA-P and PBMCs from multiple allogeneic donors irradiated at 20 Gy. Medium was replenished and topped up with IL-2 (100 IU / ml) and IL-15 (10 ng / ml) after 1 week of stimulation. After two weeks, single cell clones were stained with DP4 L112W / V141M dimers.

[0278] ELISPOT assay

[0279] Cytokine ELISPOT assays were performed as previously reported (see Yamashita, Y. et al. Nat Commun 8, 15244 (2017); Anczurowski, M. et al. Sci Rep 8, 4804 (2018)). SEQUENCE LIST <110> University Health Network <120> MHC class II molecules and methods of use thereof <130> 4285.010PC02 / C-K / BMD <150> US 62 / 880,496 <151> 30-JUL-2019 <150> US 63 / 029,111 <151> 22-MAY-2020 <160> 284 <170> PatentIn version 3.5 <210> 1 <211> 196 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 1 Arg Ala Thr Pro Glu Asn Tyr Leu Phe Gin Gly Arg Gin Glu Cys Tyr 1 5 10 15 Ala Phe Asn Gly Thr Gin Arg Phe Leu Glu Arg Tyr He Tyr Asn Arg 20 25 30 Glu Glu Phe Ala Arg Phe Asp Ser Asp Val Gly Glu Phe Arg Ala Val 35 40 45 Thr Glu Leu Gly Arg Pro Ala Ala Glu Tyr Trp Asn Ser Gin Lys Asp 50 55 60 He Leu Glu Glu Lys Arg Ala Val Pro Asp Arg Met Cys Arg His Asn 65 70 75 80 Tyr Glu Leu Gly Gly Pro Met Thr Leu Gin Arg Arg Val Gin Pro Arg 85 90 95 Val Asn Val Ser Pro Ser Lys Lys Gly Pro Leu Gin His His Asn Leu 100 105 110 Leu Val Gin His Val Thr Asp Phe Tyr Pro Gly Ser He Gin Val Arg 115 120 125 Trp Phe Leu Asn Gly Gin Gin Gin Thr Gin Gly Val Val Ser Thr Asn 130 135 140 Leu He Arg Asn Gly Asp Trp Thr Phe Gin He Leu Val Met Leu Gin 145 150 155 160 Met Thr Pro Gin Gin Gly Asp Val Tyr Thr Gin Gin Val Gin His Thr 165 170 175 Ser Leu Asp Ser Pro Val Thr Val Gin Trp Gin Ser Gin Ser Asp Ser 180 185 190 Ala Arg Ser Lys 195 <210> 2 <211> 588 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 2 Ala Gly Gly Gly Cys Cys Ala Cys Thr Cys Cys Ala Gly Ala Gly Ala 1 5 10 15 Ala Thr Thr Ala Cys Cys Thr Thr Thr Thr Cys Cys Ala Gly Gly Gly 20 25 30 Ala Cys Gly Gly Cys Ala Gly Gly Ala Ala Thr Gly Cys Thr Ala Cys 35 40 45 Gly Cys Gly Thr Thr Thr Ala Ala Thr Gly Gly Gly Ala Cys Ala Cys 50 55 60 Ala Gly Cys Gly Cys Thr Thr Cys Cys Thr Gly Gly Ala Gly Ala Gly 65 70 75 80 Ala Thr Ala Cys Ala Thr Cys Thr Ala Cys Ala Ala Cys Cys Gly Gly 85 90 95 Gly Ala Gly Gly Ala Gly Thr Thr Cys Gly Cys Gly Cys Gly Cys Thr 100 105 110 Thr Cys Gly Ala Cys Ala Gly Cys Gly Ala Cys Gly Thr Gly Gly Gly 115 120 125 Gly Gly Ala Gly Thr Thr Cys Cys Gly Gly Gly Cys Gly Gly Thr Gly 130 135 140 Ala Cys Gly Gly Ala Gly Cys Thr Gly Gly Gly Gly Cys Gly Gly Cys 145 150 155 160 Cys Thr Gly Cys Thr Gly Cys Gly Gly Ala Gly Thr Ala Cys Thr Gly 165 170 175 Gly Ala Ala Cys Ala Gly Cys Cys Ala Gly Ala Ala Gly Gly Ala Cys 180 185 190 Ala Thr Cys Cys Thr Gly Gly Ala Gly Gly Ala Gly Ala Ala Gly Cys 195 200 205 Gly Gly Gly Cys Ala Gly Thr Gly Cys Cys Gly Gly Ala Cys Ala Gly 210 215 220 Gly Ala Thr Gly Thr Gly Cys Ala Gly Ala Cys Ala Cys Ala Ala Cys 225 230 235 240 Thr Ala Cys Gly Ala Gly Cys Thr Gly Gly Gly Cys Gly Gly Gly Cys 245 250 255 Cys Cys Ala Thr Gly Ala Cys Cys Cys Thr Gly Cys Ala Gly Cys Gly 260 265 270 Cys Cys Gly Ala Gly Thr Cys Cys Ala Gly Cys Cys Thr Ala Gly Gly 275 280 285 Gly Thr Gly Ala Ala Thr Gly Thr Thr Thr Cys Cys Cys Cys Cys Thr 290 295 300 Cys Cys Ala Ala Gly Ala Ala Gly Gly Gly Gly Cys Cys Cys Thr Thr 305 310 315 320 Gly Cys Ala Gly Cys Ala Cys Cys Ala Cys Ala Ala Cys Cys Thr Gly 325 330 335 Cys Thr Thr Gly Thr Cys Thr Gly Cys Cys Ala Cys Gly Thr Gly Ala 340 345 350 Cys Gly Gly Ala Thr Thr Thr Cys Thr Ala Cys Cys Cys Ala Gly Gly 355 360 365 Cys Ala Gly Cys Ala Thr Thr Cys Ala Ala Gly Thr Cys Cys Gly Ala 370 375 380 Thr Gly Gly Thr Thr Cys Cys Thr Gly Ala Ala Thr Gly Gly Ala Cys 385 390 395 400 Ala Gly Gly Ala Gly Gly Ala Ala Ala Cys Ala Gly Cys Thr Gly Gly 405 410 415 Gly Gly Thr Cys Gly Thr Gly Thr Cys Cys Ala Cys Cys Ala Ala Cys 420 425 430 Cys Thr Gly Ala Thr Cys Cys Gly Thr Ala Ala Thr Gly Gly Ala Gly 435 440 445 Ala Cys Thr Gly Gly Ala Cys Cys Thr Thr Cys Cys Ala Gly Ala Thr 450 455 460 Cys Cys Thr Gly Gly Thr Gly Ala Thr Gly Cys Thr Gly Gly Ala Ala 465 470 475 480 Ala Thr Gly Ala Cys Cys Cys Cys Cys Cys Ala Gly Cys Ala Gly Gly 485 490 495 Gly Ala Gly Ala Thr Gly Thr Cys Thr Ala Cys Ala Cys Cys Thr Gly 500 505 510 Cys Cys Ala Ala Gly Thr Gly Gly Ala Gly Cys Ala Cys Ala Cys Cys 515 520 525 Ala Gly Cys Cys Thr Gly Gly Ala Thr Ala Gly Thr Cys Cys Thr Gly 530 535 540 Thr Cys Ala Cys Cys Gly Thr Gly Gly Ala Gly Thr Gly Gly Ala Ala 545 550 555 560 Gly Gly Cys Ala Cys Ala Gly Thr Cys Thr Gly Ala Thr Thr Cys Thr 565 570 575 Gly Cys Cys Cys Gly Gly Ala Gly Thr Ala Ala Gly 580 585 <210> 3 <211> 196 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 3 Arg Ala Thr Pro Glu Asn Tyr Leu Phe Gln Gly Arg Gln Glu Cys Tyr 1 5 10 15 Ala Phe Asn Gly Thr Gln Arg Phe Leu Glu Arg Tyr Ile Tyr Asn Arg 20 25 30 Glu Glu Phe Ala Arg Phe Asp Ser Asp Val Gly Glu Phe Arg Ala Val 35 40 45 Thr Glu Leu Gly Arg Pro Ala Ala Glu Tyr Trp Asn Ser Gln Lys Asp 50 55 60 Ile Leu Glu Glu Lys Arg Ala Val Pro Asp Arg Met Cys Arg His Asn 65 70 75 80 Tyr Glu Leu Gly Gly Pro Met Thr Leu Gln Arg Arg Val Gln Pro Arg 85 90 95 Val Asn Val Ser Pro Ser Lys Lys Gly Pro Leu Gln His His Asn Trp 100 105 110 Leu Val Cys His Val Thr Asp Phe Tyr Pro Gly Ser Ile Gln Val Arg 115 120 125 Trp Phe Leu Asn Gly Gln Glu Glu Thr Ala Gly Val Met Ser Thr Asn 130 135 140 Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val Met Leu Glu 145 150 155 160 Met Thr Pro Gln Gln Gly Asp Val Tyr Thr Cys Gln Val Glu His Thr 165 170 175 Ser Leu Asp Ser Pro Val Thr Val Glu Trp Lys Ala Gln Ser Asp Ser 180 185 190 Ala Arg Ser Lys 195 <210> 4 <211> 265 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 4 Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala Arg Ala Thr Pro Glu Asn Tyr Leu Phe Gln Gly Arg Gln Glu Cys 20 25 30 Tyr Ala Phe Asn Gly Thr Gln Arg Phe Leu Glu Arg Tyr Ile Tyr Asn 35 40 45 Arg Glu Glu Phe Ala Arg Phe Asp Ser Asp Val Gly Glu Phe Arg Ala 50 55 60 Val Thr Glu Leu Gly Arg Pro Ala Ala Glu Tyr Trp Asn Ser Gln Lys 65 70 75 80 Asp Ile Leu Glu Glu Lys Arg Ala Val Pro Asp Arg Met Cys Arg His 85 90 95 Asn Tyr Glu Leu Gly Gly Pro Met Thr Leu Gln Arg Arg Val Gln Pro 100 105 110 Arg Val Asn Val Ser Pro Ser Lys Lys Gly Pro Leu Gln His His Asn 115 120 125 Trp Leu Val Cys His Val Thr Asp Phe Tyr Pro Gly Ser lie Gin Val 130 135 140 Arg Trp Phe Leu Asn Gly Gin Glu Glu Thr Ala Gly Val Met Ser Thr 145 150 155 160 Asn Leu lie Arg Asn Gly Asp Trp Thr Phe Gin lie Leu Val Met Leu 165 170 175 Glu Met Thr Pro Gin Gin Gly Asp Val Tyr Thr Cys Gin Val Glu His 180 185 190 Thr Ser Leu Asp Ser Pro Val Thr Val Glu Trp Lys Ala Gin Ser Asp 195 200 205 Ser Ala Arg Ser Lys Gly Gly Gly Gly Ser Leu Glu lie Glu Ala Ala 210 215 220 Phe Leu Glu Arg Glu Asn Thr Ala Leu Glu Thr Arg Val Ala Glu Leu 225 230 235 240 Arg Gin Arg Val Gin Arg Leu Arg Asn Arg Val Ser Gin Tyr Arg Thr 245 250 255 Arg Tyr Gly Pro Leu Gly Gly Gly Lys 260 265 <210> 5 <211> 258 <212> PRT <213> Artificial Sequence <220> <223> Synthetic constructs <400> 5 Met Met Val Leu Gin Val Ser Ala Ala Pro Arg Thr Val Ala Leu Thr 1 5 10 15 Ala Leu Leu Met Val Leu Leu Thr Ser Val Val Gin Gly Arg Ala Thr 20 25 30 Pro Glu Asn Tyr Leu Phe Gin Gly Arg Gin Glu Cys Tyr Ala Phe Asn 35 40 45 Gly Thr Gin Arg Phe Leu Glu Arg Tyr He Tyr Asn Arg Glu Glu Phe 50 55 60 Ala Arg Phe Asp Ser Asp Val Gly Glu Phe Arg Ala Val Thr Glu Leu 65 70 75 80 Gly Arg Pro Ala Ala Glu Tyr Trp Asn Ser Gin Lys Asp He Leu Glu 85 90 95 Glu Lys Arg Ala Val Pro Asp Arg Met Cys Arg His Asn Tyr Glu Leu 100 105 110 Gly Gly Pro Met Thr Leu Gin Arg Arg Val Gin Pro Arg Val Asn Val 115 120 125 Ser Pro Ser Lys Lys Gly Pro Leu Gin His His Asn Leu Leu Val Cys 130 135 140 His Val Thr Asp Phe Tyr Pro Gly Ser He Gin Val Arg Trp Phe Leu 145 150 155 160 Asn Gly Gln Glu Glu Thr Ala Gly Val Val Ser Thr Asn Leu Ile Arg 165 170 175 Asn Gly Asp Trp Thr Phe Gln Ile Leu Val Met Leu Glu Met Thr Pro 180 185 190 Gln Gln Gly Asp Val Tyr Thr Cys Gln Val Glu His Thr Ser Leu Asp 195 200 205 Ser Pro Val Thr Val Glu Trp Lys Ala Gln Ser Asp Ser Ala Arg Ser 210 215 220 Lys Thr Leu Thr Gly Ala Gly Gly Phe Val Leu Gly Leu Ile Ile Cys 225 230 235 240 Gly Val Gly Ile Phe Met His Arg Arg Ser Lys Lys Val Gln Arg Gly 245 250 255 Ser Ala <210> 6 <211> 192 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 6 Ile Lys Ala Asp His Val Ser Thr Tyr Ala Ala Phe Val Gln Thr His 1 5 10 15 Arg Pro Thr Gly Glu Phe Met Phe Glu Phe Asp Glu Asp Glu Met Phe 20 25 30 Tyr Val Asp Leu Asp Lys Lys Glu Thr Val Trp His Leu Glu Glu Phe 35 40 45 Gly Gln Ala Phe Ser Phe Glu Ala Gln Gly Gly Leu Ala Asn Ile Ala 50 55 60 Ile Leu Asn Asn Asn Leu Asn Thr Leu Ile Gln Arg Ser Asn His Thr 65 70 75 80 Gln Ala Thr Asn Asp Pro Pro Glu Val Thr Val Phe Pro Lys Glu Pro 85 90 95 Val Glu Leu Gly Gln Pro Asn Thr Leu Ile Cys His Ile Asp Lys Phe 100 105 110 Phe Pro Pro Val Leu Asn Val Thr Trp Leu Cys Asn Gly Glu Leu Val 115 120 125 Thr Glu Gly Val Ala Glu Ser Leu Phe Leu Pro Arg Thr Asp Tyr Ser 130 135 140 Phe His Lys Phe His Tyr Leu Thr Phe Val Pro Ser Ala Glu Asp Phe 145 150 155 160 Tyr Asp Cys Arg Val Glu His Trp Gly Leu Asp Gln Pro Leu Leu Lys 165 170 175 His Trp Glu Ala Gln Glu Pro Ile Gln Met Pro Glu Thr Thr Glu Thr 180 185 190 <210> 7 <211> 576 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 7 Ala Thr Cys Ala Ala Gly Gly Cys Cys Gly Ala Cys Cys Ala Cys Gly 1 5 10 15 Thr Gly Thr Cys Cys Ala Cys Ala Thr Ala Cys Gly Cys Cys Gly Cys 20 25 30 Cys Thr Thr Cys Gly Thr Gly Cys Ala Gly Ala Cys Cys Cys Ala Cys 35 40 45 Ala Gly Ala Cys Cys Cys Ala Cys Cys Gly Gly Cys Gly Ala Gly Thr 50 55 60 Thr Cys Ala Thr Gly Thr Thr Cys Gly Ala Gly Thr Thr Cys Gly Ala 65 70 75 80 Cys Gly Ala Gly Gly Ala Cys Gly Ala Gly Ala Thr Gly Thr Thr Cys 85 90 95 Thr Ala Cys Gly Thr Gly Gly Ala Cys Cys Thr Gly Gly Ala Cys Ala 100 105 110 Ala Gly Ala Ala Ala Gly Ala Ala Ala Cys Cys Gly Thr Gly Thr Gly 115 120 125 Gly Cys Ala Cys Cys Thr Gly Gly Ala Ala Gly Ala Gly Thr Thr Cys 130 135 140 Gly Gly Cys Cys Ala Gly Gly Cys Cys Thr Thr Cys Ala Gly Cys Thr 145 150 155 160 Thr Thr Gly Ala Gly Gly Cys Cys Cys Ala Gly Gly Gly Cys Gly Gly 165 170 175 Ala Cys Thr Gly Gly Cys Cys Ala Ala Thr Ala Thr Cys Gly Cys Cys 180 185 190 Ala Thr Cys Cys Thr Gly Ala Ala Cys Ala Ala Cys Ala Ala Cys Cys 195 200 205 Thr Gly Ala Ala Cys Ala Cys Cys Cys Thr Gly Ala Thr Cys Cys Ala 210 215 220 Gly Cys Gly Gly Ala Gly Cys Ala Ala Cys Cys Ala Cys Ala Cys Cys 225 230 235 240 Cys Ala Gly Gly Cys Cys Ala Cys Cys Ala Ala Cys Gly Ala Thr Cys 245 250 255 Cys Cys Cys Cys Cys Gly Ala Ala Gly Thr Gly Ala Cys Cys Gly Thr 260 265 270 Gly Thr Thr Cys Cys Cys Cys Ala Ala Ala Gly Ala Ala Cys Cys Cys 275 280 285 Gly Thr Gly Gly Ala Ala Cys Thr Gly Gly Gly Cys Cys Ala Gly Cys 290 295 300 Cys Cys Ala Ala Thr Ala Cys Cys Cys Thr Gly Ala Thr Cys Thr Gly 305 310 315 320 Cys Cys Ala Cys Ala Thr Cys Gly Ala Cys Ala Ala Gly Thr Thr Cys 325 330 335 Thr Thr Cys Cys Cys Cys Cys Cys Cys Gly Thr Gly Cys Thr Gly Ala 340 345 350 Ala Cys Gly Thr Gly Ala Cys Cys Thr Gly Gly Cys Thr Gly Thr Gly 355 360 365 Cys Ala Ala Thr Gly Gly Cys Gly Ala Gly Cys Thr Cys Gly Thr Gly 370 375 380 Ala Cys Ala Gly Ala Gly Gly Gly Cys Gly Thr Gly Gly Cys Cys Gly 385 390 395 400 Ala Gly Thr Cys Thr Cys Thr Gly Thr Thr Cys Cys Thr Gly Cys Cys 405 410 415 Cys Ala Gly Ala Ala Cys Cys Gly Ala Cys Thr Ala Cys Ala Gly Cys 420 425 430 Thr Thr Cys Cys Ala Cys Ala Ala Gly Thr Thr Cys Cys Ala Cys Thr 435 440 445 Ala Cys Cys Thr Gly Ala Cys Cys Thr Thr Cys Gly Thr Gly Cys Cys 450 455 460 Cys Ala Gly Cys Gly Cys Cys Gly Ala Gly Gly Ala Cys Thr Thr Cys 465 470 475 480 Thr Ala Cys Gly Ala Cys Thr Gly Cys Ala Gly Ala Gly Thr Gly Gly 485 490 495 Ala Ala Cys Ala Cys Thr Gly Gly Gly Gly Cys Cys Thr Gly Gly Ala 500 505 510 Cys Cys Ala Gly Cys Cys Cys Cys Thr Gly Cys Thr Gly Ala Ala Ala 515 520 525 Cys Ala Thr Thr Gly Gly Gly Ala Ala Gly Cys Cys Cys Ala Gly Gly 530 535 540 Ala Ala Cys Cys Cys Ala Thr Cys Cys Ala Gly Ala Thr Gly Cys Cys 545 550 555 560 Cys Gly Ala Gly Ala Cys Ala Ala Cys Cys Gly Ala Gly Ala Cys Ala 565 570 575 <210> 8 <211> 269 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 8 Met Met Arg Pro lie Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala lie Lys Ala Asp His Val Ser Thr Tyr Ala Ala Phe Val Gin Thr 20 25 30 His Arg Pro Thr Gly Glu Phe Met Phe Glu Phe Asp Glu Asp Glu Met 35 40 45 Phe Tyr Val Asp Leu Asp Lys Lys Glu Thr Val Trp His Leu Glu Glu 50 55 60 Phe Gly Gin Ala Phe Ser Phe Glu Ala Gin Gly Gly Leu Ala Asn lie 65 70 75 80 Ala lie Leu Asn Asn Asn Leu Asn Thr Leu lie Gin Arg Ser Asn His 85 90 95 Thr Gin Ala Thr Asn Asp Pro Pro Glu Val Thr Val Phe Pro Lys Glu 100 105 110 Pro Val Glu Leu Gly Gin Pro Asn Thr Leu lie Cys His lie Asp Lys 115 120 125 Phe Phe Pro Pro Val Leu Asn Val Thr Trp Leu Cys Asn Gly Glu Leu 130 135 140 Val Thr Glu Gly Val Ala Glu Ser Leu Phe Leu Pro Arg Thr Asp Tyr 145 150 155 160 Ser Phe His Lys Phe His Tyr Leu Thr Phe Val Pro Ser Ala Glu Asp 165 170 175 Phe Tyr Asp Cys Arg Val Glu His Trp Gly Leu Asp Gln Pro Leu Leu 180 185 190 Lys His Trp Glu Ala Gln Glu Pro Ile Gln Met Pro Glu Thr Thr Glu 195 200 205 Thr Gly Gly Gly Gly Ser Leu Glu Ile Arg Ala Ala Phe Leu Arg Gln 210 215 220 Arg Asn Thr Ala Leu Arg Thr Glu Val Ala Glu Leu Glu Gln Glu Val 225 230 235 240 Gln Arg Leu Glu Asn Glu Val Ser Gln Tyr Glu Thr Arg Tyr Gly Pro 245 250 255 Leu Gly Gly Gly Lys Gly Ser His His His His His His 260 265 <210> 9 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <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 Construct <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 Gin Cys Arg Ser Pro Arg Gly 145 150 155 160 Lys Asn Ile Gin Gly Gly Lys Thr Leu Ser Val Ser Gin Leu Glu Leu 165 170 175 Gln Asp Ser Gly Thr Trp Thr Cys Thr Val Leu Gin Asn Gin Lys Lys 180 185 190 Val Glu Phe Lys Ile Asp Ile Val Val Leu Ala Phe Gin Lys Ala Ser 195 200 205 Ser Ile Val Tyr Lys Lys Glu Gly Glu Gin 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 Gin Asp Pro Lys Leu 260 265 270 Gln Met Gly Lys Lys Leu Pro Leu His Leu Thr Leu Pro Gin Ala Leu 275 280 285 Pro Gin Tyr Ala Gly Ser Gly Asn Leu Thr Leu Ala Leu Glu Ala Lys 290 295 300 Thr Gly Lys Leu His Gin Glu Val Asn Leu Val Val Met Arg Ala Thr 305 310 315 320 Gln Leu Gin 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 Gin Val Leu Leu Glu Ser Asn Ile 370 375 380 Lys Val Leu Pro Thr Trp Ser Thr Pro Val Gin 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 Gin Ala Glu Arg Met 420 425 430 Ser Gin Ile Lys Arg Leu Leu Ser Glu Lys Lys Thr Cys Gin Cys Pro 435 440 445 His Arg Phe Gin Lys Thr Cys Ser Pro Ile 450 455 <210> 11 <400> 11 000 <210> 12 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 12 Cys Ala Cys Cys Ala Cys Ala Ala Cys Asn Asn Asn Cys Thr Thr Asn 1 5 10 15 Asn Asn Thr Gly Cys Cys Ala Cys Gly Thr Gly 20 25 <210> 13 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 13 Cys Ala Cys Gly Thr Gly Gly Cys Ala Asn Asn Asn Ala Ala Gly Asn 1 5 10 15 Asn Asn Gly Thr Thr Gly Thr Gly Gly Thr Gly 20 25 <210> 14 <211> 27 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 14 Ala Cys Ala Gly Cys Thr Gly Gly Gly Gly Thr Cys Asn Asn Asn Thr 1 5 10 15 Cys Cys Ala Cys Cys Ala Ala Cys Cys Thr Gly 20 25 <210> 15 <211> 27 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 15 Cys Ala Gly Gly Thr Thr Gly Gly Thr Gly Gly Ala Asn Asn Asn Gly 1 5 10 15 Ala Cys Cys Cys Cys Ala Gly Cys Thr Gly Thr 20 25 <210> 16 <211> 27 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 16 Cys Ala Gly Ala Thr Cys Asn Asn Asn Gly Thr Gly Asn Asn Asn Cys 1 5 10 15 Thr Gly Gly Ala Ala Ala Thr Gly Ala Cys Cys 20 25 <210> 17 <211> 27 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 17 Gly Gly Thr Cys Ala Thr Thr Thr Cys Cys Ala Gly Asn Asn Asn Cys 1 5 10 15 Ala Cys Asn Asn Asn Gly Ala Thr Cys Thr Gly 20 25 <210> 18 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 18 Cys Ala Val Cys Thr Leu Tyr Asn Phe Asn Lys Phe Tyr Phe 1 5 10 <210> 19 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 19 Cys Ala Val Ala Pro Tyr Asp Arg Gly Ser Thr Leu Gly Arg Leu Tyr 1 5 10 15 Phe <210> 20 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 20 Cys Ala Gly Val Lys Asp Ser Asn Tyr Gln Leu Ile Trp 1 5 10 <210> 21 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 21 Cys Ala Leu Ser Asp Leu Ser Tyr Gly Gly Ala Thr Asn Lys Leu Ile 1 5 10 15 Phe <210> 22 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 22 Cys Ala Val Glu Val Asn Ser Gly Thr Tyr Lys Tyr Ile Phe 1 5 10 <210> 23 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 23 Cys Ala Leu Ser Val Gly Thr Tyr Lys Tyr Ile Phe 1 5 10 <210> 24 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 24 Cys Ala Phe Met Lys Arg Ala Glu Thr Ser Gly Ser Arg Leu Thr Phe 1 5 10 15 <210> 25 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 25 Cys Ala Ser Leu Thr Asp Asn Asn Glu Gin Phe Phe 1 5 10 <210> 26 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 26 Cys Ala Ser Ser Thr Gly Gin Gly Leu Glu Thr Gin Tyr Phe 1 5 10 <210> 27 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 27 Cys Ala Trp Ser Ser Tyr Asn Glu Gin Phe Phe 1 5 10 <210> 28 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 28 Cys Ala Ser Ser Lys Gly Gin Gly Leu Gly Asn Gin Pro Gin His Phe 1 5 10 15 <210> 29 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 29 Cys Ala Ser Arg Arg Asp Leu Ala Ala Phe Phe 1 5 10 <210> 30 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 30 Cys Ala Ser Ser Pro Gly Thr Gly Gly Arg Glu Thr Gin Tyr Phe 1 5 10 15 <210> 31 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 31 Cys Ala Ser Ser Tyr Trp Pro Thr Arg Glu Thr Gin Tyr Phe 1 5 10 <210> 32 <211> 20 <212> PRT <213> Artificial Sequence <220> ​​​​<223> SYNTHETIC CONSTRUCT <400> 32 Glu Val Leu Ala Pro Val lie Leu Met Leu Leu Asn Ser Phe Phe Asn 1 5 10 15 Ala lie Ser Thr 20 <210> 33 <211 > 20 <212> PRT <213> Artificial Sequence <220> <223> SYNTHETIC CONSTRUCT <400> 33 Glu Gly Glu lie Ser Asp Pro Phe Arg Phe Thr Thr Phe Tyr lie His 1 5 10 15 Phe Ala Leu Val 20 <210> 34 <211 > 20 <212> PRT <213> Artificial Sequence <220> <223> SYNTHETIC CONSTRUCT <400> 34 Lys Ala Leu Leu Ala Thr Phe Gly Ser Ser Phe Leu lie Ser Ala Cys 1 5 10 15 Phe Lys Leu lie 20 <210> 35 <211 > 20 <212> PRT <213> Artificial Sequence <220> <223> SYNTHETIC CONSTRUCT <400> 35 Phe Lys Leu lie Gin Asp Leu Leu Ser Phe lie Asn Pro Gin Leu Leu 1 5 10 15 Ser lie Leu lie 20 <210> 36 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 36 Gly Cys Glu Asp Val Arg Val Ser Gly Trp Leu Gin Thr Glu Phe Leu 1 5 10 15 Ser Phe Gin Asp 20 <210> 37 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 37 Thr Val Thr Lys Leu Ser Gin Lys Phe Thr Lys Val Asn Phe Thr Glu 1 5 10 15 lie Gin Lys Leu 20 <210> 38 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 38 Thr Asp Glu Glu Leu Asp Arg Phe Lys Phe Phe Leu Ser Asp Glu Phe 1 5 10 15 Asn Ile Ala Thr 20 <210> 39 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 39 Arg Ile Ile Ile Ile Ala Arg Tyr Tyr Arg His Ser Gly Phe Leu Glu 1 5 10 15 Val Asn Ser Ala 20 <210> 40 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 40 Arg Thr Ile Pro Ile Asp Gly Asn Phe Phe Thr Tyr Thr Arg His Glu 1 5 10 15 Pro Ile Gly Val 20 <210> 41 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 41 Asp Glu Leu Asp Phe Leu Met Glu Ala Leu Ile Ile Ser Lys Phe Asn 1 5 10 15 His Gln Asn Ile 20 <210> 42 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 42 Val Tyr Gly Asn Thr Ala Leu His Tyr Ala Val Tyr Ser Glu Ile Leu 1 5 10 15 Ser Val Val Ala 20 <210> 43 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 43 Leu Glu Ser Ile Arg Lys Glu Val Lys Gly Asp Leu Glu Asn Ala Phe 1 5 10 15 Leu Asn Leu Val 20 <210> 44 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 44 Arg Ile Arg Pro Leu Trp Lys His Tyr Phe Gln Asn Thr Gln Gly Leu 1 5 10 15 Ile Phe Val Val 20 <210> 45 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 45 Ala Val Phe Leu Ala Leu Ser Ala Gin Leu Leu Gin Ala Arg Leu Met 1 5 10 15 Lys Glu Glu Ser 20 <210> 46 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 46 Arg Thr He Met Gly Trp Thr Leu Asp Phe Leu Arg Glu Arg Leu Leu 1 5 10 15 Gly Trp He Gin 20 <210> 47 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 47 Leu Ser Tyr Lys Leu Ser Gin Lys Gly Tyr Ser Trp Ser Gin Phe Ser 1 5 10 15 Asp Val Glu Glu 20 <210> 48 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 48 Leu Ser Val Lys Lys Gin Phe Glu Glu Leu Thr Leu Gly Glu Phe Leu 1 5 10 15 Lys Leu Asp Arg 20 <210> 49 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 49 Asp Pro Trp Thr Glu His Ala Lys Trp Phe Pro Ser Cys Gin Phe Leu 1 5 10 15 Leu Arg Ser Lys 20 <210> 50 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 50 Tyr Pro Ser Ser Gin Asp Ser Ser Ser Ala Ala Ala Pro Gin Leu Leu 1 5 10 15 Ile Val Leu Leu 20 <210> 51 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 51 Leu Thr Thr Pro Pro Cys Ala Gin Gly Val He Trp Thr Val Phe Asn 1 5 10 15 Gln Thr Val Met 20 <210> 52 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 52 Met Gly Phe Gin Lys Phe Ser Pro Phe Leu Ala Leu Ser He Leu Val 1 5 10 15 Leu Leu Gin Ala 20 <210> 53 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 53 Val Leu Gin Gin Gin Leu Glu Ser Phe Gin Ala Leu Arg Met Gin Thr 1 5 10 15 Leu Gin Asn Val 20 <210> 54 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 54 Lys Tyr Val Ala Glu Leu Ser Leu Leu Glu Ala Asp Pro Phe Leu Lys 1 5 10 15 Tyr Leu Pro Ser 20 <210> 55 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 55 Gln Gln Ala Ile Arg Glu Lys Tyr Lys Ala Ser Lys Tyr Leu Cys Val 1 5 10 15 Ser Leu Met Glu 20 <210> 56 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 56 Ser Pro Asn Asn Phe Leu Ser Tyr Tyr Arg Leu Thr Arg Phe Leu Ser 1 5 10 15 Arg Val Ile Lys 20 <210> 57 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 57 Met Arg Ile Phe Ala Val Phe Ile Phe Met Thr Tyr Trp His Leu Leu 1 5 10 15 Asn Ala Phe Thr 20 <210> 58 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 58 Pro Ser Lys Tyr lie Asn Ala Ser Phe lie Met Ser Tyr Trp Lys Pro 1 5 10 15 Glu Val Met lie 20 <210> 59 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 59 Pro Leu Lys Glu Thr lie Gly Asp Phe Trp Gin Met lie Phe Gin Arg 1 5 10 15 Lys Val Lys Val 20 <210> 60 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 60 Lys Ala Arg Pro Gly Met Val Ser Thr Phe Glu Gin Tyr Gin Phe Leu 1 5 10 15 Tyr Asp Val lie 20 <210> 61 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 61 Asp Phe Val Thr Glu Thr Pro Leu Glu Gly Asp Phe Ala Trp Glu Arg 1 5 10 15 Val Arg Gly Leu 20 <210> 62 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 62 Asp Phe Val Thr Glu Thr Pro Leu Glu Gly Asp Phe Ala Trp Glu Arg 1 5 10 15 Val Arg Gly Leu 20 <210> 63 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 63 Pro Ala Gln Tyr Ser Trp Phe Val Asn Gly Thr Phe Gln Gln Ser Thr 1 5 10 15 Gln Glu Leu Phe 20 <210> 64 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 64 Arg Ser Leu Arg Thr Asn Phe Leu Arg Tyr Trp Thr Leu Thr Tyr Leu 1 5 10 15 Ala Leu Pro Thr 20 <210> 65 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 65 Cys Gly Lys Glu Gly Lys Tyr Ile His Phe Thr Pro Asn Phe Leu Leu 1 5 10 15 Asn Asp Asn Leu 20 <210> 66 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 66 Ile Ser Arg Ser Asp Glu Gly Lys Tyr Thr Cys Phe Ala Glu Asn Phe 1 5 10 15 Met Gly Lys Ala 20 <210> 67 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 67 Gln Gln Cys Thr Asp Asp Val Arg Leu Phe Ala Phe Val Arg Phe Thr 1 5 10 15 Thr Gly Asp Ala 20 <210> 68 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 68 Asn Ile Ile Asp Leu Gln Phe Leu His Gly Tyr Tyr Glu Pro Thr Leu 1 5 10 15 Leu Ile Leu Phe 20 <210> 69 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 69 Ala Asn Ala Ala Val Gly Glu Pro Ala Phe Leu Ser Glu Glu Phe Gln 1 5 10 15 Asn Ser Pro Glu 20 <210> 70 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 70 Gly Val Lys Arg Lys Asp Gln Gly Phe Leu Glu Lys Glu Phe Tyr His 1 5 10 15 Lys Thr Asn Ile 20 <210> 71 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 71 His Asp Lys Ala Cys Val Arg Thr Phe Tyr Glu Thr Pro Leu Gln Leu 1 5 10 15 Leu Glu Lys Val 20 <210> 72 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 72 Gln Arg Ser Glu His Asp Val Leu Phe Gln Val Thr Gln Phe Pro Ser 1 5 10 15 Arg Gly Gln Leu 20 <210> 73 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 73 Phe Gln Ile Asp Gln Gly Glu Val Val Phe Ala Phe Thr Asn Phe Ser 1 5 10 15 Ser Ser His Asp 20 <210> 74 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 74 Ile Asp Gin Gly Glu Val Val Phe Ala Phe Thr Asn Phe Ser Ser Ser 1 5 10 15 His Asp His Phe 20 <210> 75 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 75 Ser Ser Ile Leu Cys Ala Leu Ile Val Phe Trp Lys Tyr Arg Arg Phe 1 5 10 15 Gln Arg Asn Thr 20 <210> 76 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 76 Ala Gly Gin Ser Arg Cys Gly Gly Phe Leu Val Arg Glu Asp Phe Val 1 5 10 15 Leu Thr Ala Ala 20 <210> 77 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 77 Glu Gin Leu Asn Arg Asn Phe Ser Asn Phe He Leu Asp Lys Phe Leu 1 5 10 15 Arg His Cys Glu 20 <210> 78 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 78 Ser Thr Ala Leu Gin Trp Leu Leu Leu Leu Phe Thr Arg Tyr Pro Asp 1 5 10 15 Val Gin Thr Arg 20 <210> 79 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 79 Asp Pro Trp Pro Leu Asn Pro Leu Ser He Gin Gin Thr Thr Leu Leu 1 5 10 15 Leu Leu Leu Ser 20 <210> 80 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 80 Pro Gin Phe Ala Asn Cys Ser Val Tyr Asp Phe Phe Val Trp Leu His 1 5 10 15 Tyr Tyr Ser Val 20 <210> 81 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 81 Ala Asn Cys Ser Val Tyr Asp Phe Phe Val Trp Leu His Tyr Tyr Ser 1 5 10 15 Val Arg Asp Thr 20 <210> 82 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 82 Gln Lys Phe Asp Asn Pro Pro Phe Phe Gin Asn Ser Thr Phe Ser Phe 1 5 10 15 Arg Asn Ala Leu 20 <210> 83 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 83 Ile Ala Thr Pro Gly Arg Leu Asn Asp Leu Gin Met Ser Asn Phe Val 1 5 10 15 Asn Leu Lys Asn 20 <210> 84 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 84 Cys Ala Ser Gly Leu Cys Cys Ala Arg His Phe Trp Ser Lys Ile Cys 1 5 10 15 Lys Pro Val Leu 20 <210> 85 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 85 Glu Glu Gly Cys Glu Ala Ile Ser Phe Leu Leu Ser Leu Ile Asp Arg 1 5 10 15 Leu Val Leu Tyr 20 <210> 86 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 86 Gly Ser Lys Glu Asp Ala Asn Val Phe Ala Ser Ala Met Met His Ala 1 5 10 15 Leu Glu Val Leu 20 <210> 87 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 87 Thr Arg Tyr Gin Leu Asp Pro Lys Phe He Thr Ser He Leu Tyr Glu 1 5 10 15 Asn Asn Val He 20 <210> 88 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 88 Glu Ser Asp Leu Asp Tyr Gly Thr Asn Phe Gin Lys Arg Leu Phe Thr 1 5 10 15 Lys He Asp Thr 20 <210> 89 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 89 Glu Ser Asp Asp Asp His Gly Val Lys Phe Arg Glu His Gin Phe Thr 1 5 10 15 Lys He Asp Thr 20 <210> 90 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 90 Pro Glu Gly Ala Ser Ala Ala Ser Phe Glu Tyr Thr Ile Leu Asp Pro 1 5 10 15 Ser Ser Gln Leu 20 <210> 91 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 91 Glu Asp Leu Gly Pro Ala Ser Pro Leu Asp Ser Thr Phe Tyr Arg Ser 1 5 10 15 Leu Leu Glu Asp 20 <210> 92 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 92 Asp Leu Gly Pro Ala Ser Pro Leu Asp Ser Thr Phe Tyr Arg Ser Leu 1 5 10 15 Leu Glu Asp Asp 20 <210> 93 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 93 Ala Ala Ser Gin His Val Phe Arg Phe Tyr Arg Gin Ser Leu Gin Arg 1 5 10 15 Arg Tyr Ser Lys 20 <210> 94 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 94 His Thr Leu Phe Cys Arg Arg Cys Phe Lys Tyr Asp Cys Phe Leu His 1 5 10 15 Pro Phe His Ala 20 <210> 95 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 95 Met Ser Leu Ser Phe Leu Leu Leu Leu Phe Phe Ser His Leu Ile Leu 1 5 10 15 Ser Ala Trp Ala 20 <210> 96 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 96 Val Tyr Glu Thr Tyr Glu Leu Val Glu Lys Phe Tyr Asp Pro Met Phe 1 5 10 15 Lys Tyr His Leu 20 <210> 97 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 97 Val Ala Gln Thr Gly Ile Leu Trp Leu Leu Met Asn Asn Cys Phe Leu 1 5 10 15 Asn Leu Ser Pro 20 <210> 98 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 98 Met Asn Leu Ser Arg Gly Lys Ser Thr Tyr Tyr Trp Pro Arg Pro Arg 1 5 10 15 Arg Tyr Val Gln 20 <210> 99 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 99 Met Lys Gln Asp Phe Ser Val Pro Gln Leu Pro His Ser Ser Ser His 1 5 10 15 Trp Leu Arg Leu 20 <210> 100 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 100 Pro Ser Leu Thr Pro Gln Ala Phe Glu Phe Val Gly Glu Phe Phe Thr 1 5 10 15 Asp Val Ser Leu 20 <210> 101 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 101 Ala Met Trp Ile Gln Leu Leu Tyr Ser Ala Cys Phe Trp Trp Leu Phe 1 5 10 15 Cys Tyr Ala Val 20 <210> 102 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 102 Thr Gly Pro lie Arg Asn Gly Asp Trp Thr Phe Gin Thr Val Val Met 1 5 10 15 Leu Glu Met Thr 20 <210> 103 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 103 Pro Gly Val Tyr Thr Lys Val Ser Asp Phe Arg Glu Trp lie Phe Gin 1 5 10 15 Ala lie Lys Thr 20 <210> 104 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 104 Gly Thr Val Ala Tyr Leu Ala Leu Arg lie Ser Tyr Ser Leu Phe Thr 1 5 10 15 Ala Leu Arg Val 20 <210> 105 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 105 Val Phe Val Gln Ser Val Leu Pro Tyr Phe Val Ala Thr Lys Leu Ala 1 5 10 15 Lys Ile Arg Lys 20 <210> 106 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 106 Glu Gly Ile Asp Phe Tyr Thr Ser Ile Thr Arg Ala Arg Phe Glu Glu 1 5 10 15 Leu Cys Ser Asp 20 <210> 107 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 107 Ser Phe Arg Asp Gly Asp Cys Ser Lys Gly Phe Phe Leu Val Ser Leu 1 5 10 15 Leu Val Glu Ile 20 <210> 108 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 108 Ser Ser Glu Asn Lys Pro lie Arg Ser Ser Tyr Phe Thr Phe Gin Leu 1 5 10 15 Gln Asn lie Val 20 <210> 109 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 109 Asp Asp Ala Ala Pro Arg Val Glu Gly Val Pro Val Ala Val His Lys 1 5 10 15 His Ala Leu His 20 <210> 110 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 110 Thr Phe Gly Glu Met Ala Asp Ala Phe Lys Ser Asp Tyr Phe Asn Met 1 5 10 15 Pro Val His Met 20 <210> 111 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 111 Ile Ile Ile Leu Val Gly Thr Ala Val Ile Ala Met Phe Phe Trp Leu 1 5 10 15 Leu Leu Val Ile 20 <210> 112 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 112 Arg Phe Ser Ile Trp Ile Ser Phe Phe Glu Ile Tyr Asn Glu Leu Leu 1 5 10 15 Tyr Asp Leu Leu 20 <210> 113 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 113 Asn Gln Pro Cys Tyr Arg Lys Leu Gly Leu Glu Val Tyr Val Thr Phe 1 5 10 15 Phe Glu Ile Tyr 20 <210> 114 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 114 Ser Val Arg His Pro Glu Tyr Asn Arg Pro Leu Leu Ala Asn Asp Leu 1 5 10 15 Met Leu Ile Lys 20 <210> 115 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 115 Gln Lys Lys Thr Leu Gln Ala Leu Glu Phe His Thr Val Pro Phe Gln 1 5 10 15 Leu Leu Ala Arg 20 <210> 116 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 116 Val Met Val Asn Gly Ile Leu Phe Val Gln Tyr Phe His Arg Val Pro 1 5 10 15 Phe His Arg Val 20 <210> 117 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 117 Ile Ala Lys Arg Gin Leu Ser His Leu Gin Ala Ser Gly Phe Ser Leu 1 5 10 15 Leu Ser Ala Phe 20 <210> 118 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic Construct <400> 118 Thr Gly Val Lys Glu Val Ala Arg Lys Tyr Asn Tyr Ile Ala Ser Phe 1 5 10 15 Phe Ile Glu Thr 20 <210> 119 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic Construct <400> 119 Tyr Asn Tyr Ile Ala Ser Phe Phe Ile Glu Thr Gly Phe Cys Asp Ser 1 5 10 15 Gly Ile Leu Ser 20 <210> 120 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic Construct <400> 120 Gln Ala Met Ala Lys Asn Arg Leu Gln Phe Val Arg Phe Glu Ala Thr 1 5 10 15 Asp Leu His Gly 20 <210> 121 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic constructs <400> 121 Gln Cys Phe Val Cys Ala Gln Cys Phe Gln Gln Phe Pro Glu Gly Leu 1 5 10 15 Phe Tyr Glu Phe 20 <210> 122 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic constructs <400> 122 Glu Lys Arg Phe Leu Leu Glu Glu Pro Met Pro Phe Phe Tyr Leu Lys 1 5 10 15 Cys Cys Lys Ile 20 <210> 123 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic constructs <400> 123 Asp Glu Lys Val Thr Asp Leu Val Gin Phe Leu Leu Phe Lys Tyr Gin 1 5 10 15 Met Lys Glu Pro 20 <210> 124 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 124 Ala Leu Ser Arg Lys Met Ala Glu Leu Val His Phe Leu Leu Leu Lys 1 5 10 15 Tyr Arg Ala Arg 20 <210> 125 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 125 Pro Arg Lys Leu Leu Thr Gin Asp Leu Val Gin Glu Lys Tyr Leu Glu 1 5 10 15 Tyr Arg Gin Val 20 <210> 126 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 126 Ala lie Ser Arg Lys Met Val Glu Leu Val His Phe Leu Leu Leu Lys Tyr Arg Ala Arg 1 5 10 15 Tyr Arg Ala Arg 20 <210> 127 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 127 Arg Lys Met Val Glu Leu Val His Phe Leu Leu Leu Lys Tyr Arg Ala Arg 1 5 10 15 Arg Glu Pro Val 20 <210> 128 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 128 Tyr Glu Phe Leu Trp Gly Pro Arg Ala Leu Ala Glu Thr Ser Tyr Val 1 5 10 15 Lys Val Leu Glu 20 <210> 129 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 129 Leu Gly Ser Val Val Gly Asn Trp Gin Tyr Phe Phe Pro Val He Phe Gly Lys Ala Ser Glu Phe Met Gin Val He Ser Ser Leu Pro Arg Leu Val Thr Val Ser Ser 1 5 10 15 Ser Lys Ala Ser 20 <210> 130 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 130 Pro Lys Lys Leu Leu Thr Gin Tyr Phe Val Gin Glu Asn Tyr Leu Glu 1 5 10 15 Tyr Arg Gin Val 20 <210> 131 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 131 Tyr Phe Pro Val He Phe Gly Lys Ala Ser Glu Phe Met Gin Val He Ser Ser Leu Pro Arg Leu Val Thr Val Ser Ser 1 5 10 15 Phe Gly Thr Asp 20 <210> 132 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 132 Pro Arg Lys Phe lie Thr Gin Asp Leu Val Gin Glu Lys Tyr Leu Lys 1 5 10 15 Tyr Glu Gin Val 20 <210> 133 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 133 Pro Trp Lys Leu lie Thr Lys Asp Leu Val Gin Glu Lys Tyr Leu Glu 1 5 10 15 Tyr Lys Gin Val 20 <210> 134 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 134 Gln Ser Pro Leu Gin Asn Pro Ala Ser Ser Phe Phe Ser Ser Ala Leu 1 5 10 15 Leu Ser lie Phe 20 <210> 135 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 135 Gln Ser Pro Leu Gln lie Pro Val Ser Arg Ser Phe Ser Ser Thr Leu 1 5 10 15 Leu Ser lie Phe 20 <210> 136 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 136 Ser Pro Leu Gln lie Pro Gly Ser Pro Ser Phe Ser Ser Thr Leu Leu 1 5 10 15 Ser Leu Phe Gln 20 <210> 137 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 137 Ser Pro Leu Gln lie Pro Met Thr Ser Ser Phe Ser Ser Thr Leu Leu 1 5 10 15 Ser lie Leu Gln 20 <210> 138 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 138 Pro Leu Ser Ser Cys Cys Ser Ser Phe Ser Trp Ser Ser Phe Ser Glu 1 5 10 15 Glu Ser Ser Ser 20 <210> 139 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 139 Ile Leu Thr Val Ile Leu Gly Val Leu Leu Leu Ile Gly Cys Trp Tyr 1 5 10 15 Cys Arg Arg Arg 20 <210> 140 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 140 Ala Val Asp Arg Tyr Ile Ser Ile Phe Tyr Ala Leu Arg Tyr His Ser 1 5 10 15 Ile Val Thr Leu 20 <210> 141 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 141 Ile Leu Leu Gly lie Phe Phe Leu Cys Trp Gly Pro Phe Phe Leu His 1 5 10 15 Leu Thr Leu lie 20 <210> 142 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 142 Met Gin His Arg Gly Phe Leu Leu Leu Thr Leu Leu Ala Leu Leu Ala 1 5 10 15 Leu Thr Ser Ala 20 <210> 143 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 143 Thr Tyr Thr Met Lys Glu Val Leu Phe Tyr Leu Gly Gin Tyr lie Met 1 5 10 15 Thr Lys Arg Leu 20 <210> 144 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 144 Val Ser Arg lie Ser Ala lie Phe Ser Thr Phe lie Gly Glu His Tyr 1 5 10 15 Val His Val Asn 20 <210> 145 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 145 Arg Ser Ala Met Cys Ala Phe Pro lie Lys Tyr Val Asn Asp Phe Phe 1 5 10 15 Asn Lys lie Val 20 <210> 146 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 146 Lys Leu Gly Phe Phe Leu Val Thr Phe Gly Phe lie Trp Gly Met Met 1 5 10 15 Leu Leu His Phe 20 <210> 147 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 147 Ala Pro Gin Glu Glu Lys Ala Val Phe Phe Ala Gly Asn Glu Tyr Trp 1 5 10 15 Arg Thr Val Thr 20 <210> 148 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 148 Ala Pro Gin Glu Glu Lys Ala Val Phe Phe Ala Gly Asn Glu Tyr Trp 1 5 10 15 Ile Tyr Ser Ala 20 <210> 149 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 149 Ser Lys Asn Lys Lys Thr Tyr Ile Phe Ala Gly Asp Lys Phe Trp Arg 1 5 10 15 Tyr Asn Glu Val 20 <210> 150 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 150 Leu Gin Gly Gly Gly His Ser Tyr Phe Phe Lys Gly Ala Tyr Tyr Leu 1 5 10 15 Lys Leu Glu Asn 20 <210> 151 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 151 Glu Leu Gin Trp Glu Gin Ala Gin Asp Tyr Leu Lys Arg Phe Tyr Leu 1 5 10 15 Tyr Asp Ser Glu 20 <210> 152 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 152 Gln Pro Tyr Thr Glu Tyr He Ser Thr Arg Trp Tyr Arg Ala Pro Glu 1 5 10 15 Cys Leu Leu Thr 20 <210> 153 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 153 Lys Gly Arg Val Gly Pro Leu Leu Ala Cys Ile Ile Gly Thr Gin Phe 1 5 10 15 Arg Lys Leu Arg 20 <210> 154 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 154 Asp Pro Gly Ala Ala Glu Gin Val Pro Asp Phe Val Thr Phe Leu Tyr 1 5 10 15 Gin Ile Thr Arg 20 <210> 155 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 155 Gin Met Asp Arg Val Asn Ala Ile Pro Phe Thr Tyr Glu Gin Leu Asp 1 5 10 15 Val Leu Lys His 20 <210> 156 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 156 ​​​​​​​​​​​​​​​​​​​​​​​​​​​​​Gln Leu Ser Thr Gly Val Ser Phe Phe Phe Leu Ser Phe His Ile Ser 1 5 10 15 Asn Leu Gln Phe 20 <210> 157 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 157 Ser Met Pro Ala Asn Phe Glu Thr Thr Gly Phe Glu Ala Glu Pro Phe 1 5 10 15 Ser His Leu Thr 20 <210> 158 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 158 Ser Leu Pro Ser Ser Thr Pro Val Pro Phe Ser Ser Ser Thr Phe Thr 1 5 10 15 Thr Thr Asp Ser 20 <210> 159 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 159 Ala Lys Thr Thr Thr Thr Phe Asn Thr Leu Ala Gly Ser Leu Phe Thr 1 5 10 15 Pro Leu Thr Thr 20 <210> 160 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 160 Ser Thr Lys Ala Ile Ser Ala Ser Ser Phe Gln Ser Thr Gly Phe Thr 1 5 10 15 Glu Thr Pro Glu 20 <210> 161 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 161 Ile Leu Leu Thr Ile Lys Asp Asp Thr Ile Tyr Leu Thr Arg His Leu 1 5 10 15 Ala Val Leu Asn 20 <210> 162 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 162 Ser Gly Trp Gly Asp Pro His Tyr lie Thr Phe Asp Gly Thr Tyr Tyr 1 5 10 15 Thr Phe Leu Asp 20 <210> 163 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 163 Met Arg Ala Leu Gin lie Val Tyr Gly lie Arg Leu Glu His Phe Tyr 1 5 10 15 Met Met Pro Val 20 <210> 164 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 164 Leu Val Met Gly Val Asp Val Met Phe lie Ser Leu Ser Tyr Phe Leu 1 5 10 15 lie lie Arg Thr 20 <210> 165 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 165 Asp Pro Ser Val Glu Pro Pro Leu Ser Gin Glu Thr Phe Ser Asp Leu 1 5 10 15 Trp Lys Leu Leu 20 <210> 166 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 166 Gln lie Ala Ala Val Cys Arg Glu Cys Leu Gin Ala Leu Glu Phe Leu 1 5 10 15 His Ala Asn Gin 20 <210> 167 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 167 Val Glu Lys Arg Gly Ser Ala Lys Glu Leu Leu Gin His Pro Phe Leu 1 5 10 15 Lys Leu Ala Lys 20 <210> 168 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 168 Pro Arg His Val Asp Arg Ser Asp Phe Phe Thr Ser Phe Tyr Asp Lys 1 5 10 15 Leu Lys Leu Gln 20 <210> 169 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 169 Met Phe Val Asp Ser Asp Ser Thr Tyr Cys Ser Ser Thr Val Phe Leu 1 5 10 15 Asp Thr Met Pro 20 <210> 170 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 170 Arg Lys Gln Arg Arg Ser Arg Thr Thr Phe Thr Ala Glu Gln Leu Glu 1 5 10 15 Glu Leu Glu Arg 20 <210> 171 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 171 Leu Thr Gly Met Val Pro Gly Ser Glu Phe Ser Gly Ser Pro Tyr Ser 1 5 10 15 His Pro Gin Tyr 20 <210> 172 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 172 Val Trp Asn Gly Pro Val Gly Val Phe Glu Trp Glu Ala Phe Ala Arg 1 5 10 15 Gly Thr Lys Ala 20 <210> 173 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 173 Gln Ala Gly Ala Gin Glu Ala Gin Pro Leu Gin Pro Ser His Phe Leu 1 5 10 15 Asp He Ser Glu 20 <210> 174 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 174 Asn Leu Pro Leu Val Ser Ser Thr Tyr Asp Leu Met Ser Ser Ala Tyr 1 5 10 15 Leu Ser Thr Lys 20 <210> 175 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 175 Asn Glu Arg Phe Arg Cys Pro Glu Ala Leu Phe Gln Pro Cys Phe Leu 1 5 10 15 Gly Met Glu Ser 20 <210> 176 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 176 Leu Ala Thr Gly Glu Lys Gly Phe Gly Tyr Lys Asn Ser Lys Phe His 1 5 10 15 Arg Val Ile Lys 20 <210> 177 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 177 Ser Leu Gin Cys Leu Gin Ala Leu Tyr Val Asp Ser Leu Phe Phe Leu 1 5 10 15 Arg Gly Arg Leu 20 <210> 178 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 178 Tyr Gin Cys Lys Val Cys Pro Ala Lys Phe Thr Gin Phe Val His Leu 1 5 10 15 Lys Leu His Lys 20 <210> 179 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 179 Glu Val Ala Ala Lys Thr Leu Pro Phe Tyr Lys Asp Tyr Phe Asn Val 1 5 10 15 Pro Tyr Pro Leu 20 <210> 180 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 180 Lys Leu Asn Leu Gly Thr Val Gly Phe Tyr Arg Thr Gin Tyr Ser Ser 1 5 10 15 Ala Met Leu Glu 20 <210> 181 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 181 Ser His Thr Asp Phe Tyr Glu Glu lie Gin Glu Phe Val Lys Asp Val 1 5 10 15 Phe Ser Pro lie 20 <210> 182 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 182 Glu Leu Tyr Asn Arg Tyr Gin Gly Gly Phe Leu lie Ser Arg Leu lie 1 5 10 15 Lys Leu Ser Val 20 <210> 183 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 183 Cys Lys Leu Ser Ser Ala Arg Trp Gly Val Cys Trp Val Asn Phe Glu 1 5 10 15 Ala Leu Ile Ile 20 <210> 184 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 184 Ser Arg Leu Gly Pro Thr Leu Met Cys Leu Leu Ser Thr Gln Phe Lys 1 5 10 15 Arg Leu Arg Asp 20 <210> 185 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 185 Gln Gly Lys Asp Val Leu Met Asn Asn Gly Leu Lys Met Asp Gln Phe 1 5 10 15 Cys Lys Glu His 20 <210> 186 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 186 Asn Thr Lys Phe Ser Glu Glu Glu Leu Cys Ser Trp Tyr Gln Ser Phe 1 5 10 15 Leu Lys Asp Cys 20 <210> 187 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 187 Met Glu Lys Asp Ser Leu Pro Arg Phe Val Arg Ser Glu Phe Tyr Gln 1 5 10 15 Glu Leu Ile Lys 20 <210> 188 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 188 Tyr Gly Leu Ala Ser Phe Lys Ser Phe Leu Lys Ser Glu Phe Ser Glu 1 5 10 15 Glu Asn Leu Glu 20 <210> 189 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 189 ​​​Cys Gly Lys Thr Cys Leu Leu Ile Val Phe Ser Lys Asp Gln Phe Pro 1 5 10 15 Glu Val Tyr Val 20 <210> 190 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 190 Pro Asp Tyr Asp Val Trp Ile Leu Met Thr Val Val Gly Thr Ile Phe 1 5 10 15 Val Ile Ile Leu 20 <210> 191 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 191 Tyr Leu Thr Pro Asp Leu Trp Lys Glu Thr Val Phe Thr Lys Ser Pro 1 5 10 15 Tyr Gln Glu Phe 20 <210> 192 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 192 Lys Val Lys Arg Gin Phe Val Glu Phe Thr lie Lys Glu Ala Ala Arg 1 5 10 15 Phe Lys Lys Val 20 <210> 193 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 193 Met Asp Gin Glu Phe Gly Val Ser Thr Leu Val Glu Glu Glu Phe Gly 1 5 10 15 Gln Arg Arg Gin 20 <210> 194 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 194 Arg Gin Lys Met Ser Glu lie Phe Pro Leu Thr Glu Glu Leu Trp Leu 1 5 10 15 Glu Trp Leu His 20 <210> 195 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 195 Leu Ser Asn Val Glu Val Phe Met Val Ile Ser Phe Ser Ser Tyr Lys 1 5 10 15 Leu Phe Lys Ser 20 <210> 196 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 196 Thr Leu Arg Asp Lys Ala Ser Gly Val Cys Ile Asp Ser Glu Phe Phe 1 5 10 15 Leu Thr Thr Ala 20 <210> 197 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 197 Val Gly Leu Ile Phe Ala Val Cys Leu Val Gly Phe Met Leu Tyr Arg 1 5 10 15 Met Lys Lys Lys 20 <210> 198 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 198 His His His Leu Leu Gin Glu Tyr Glu lie Glu Arg Ser Phe Phe Leu 1 5 10 15 Arg Met Lys Cys 20 <210> 199 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 199 Leu Leu Val Pro Leu Leu Leu Ser Leu Phe Val Leu Gly Leu Phe Leu 1 5 10 15 Trp Phe Leu Lys 20 <210> 200 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 200 Val Gin Arg Leu Trp Val Ser Arg Leu Leu Arg His Arg Lys Ala Gin 1 5 10 15 Leu Leu Leu Val 20 <210> 201 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 201 Ser Gin lie Ala Tyr Thr Ser Leu Ser Leu Pro His Tyr Gly Ser Ala 1 5 10 15 Phe Pro Ser lie 20 <210> 202 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 202 Asn Phe Glu Ser Met Ser Leu Gly Ser Phe Ser Ser Ser Ser Ala Leu 1 5 10 15 Asp Arg Asp Leu 20 <210> 203 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 203 Arg Glu Gin Pro Asp Asn lie Pro Ala Phe Ala Ala Ala Tyr Phe Glu 1 5 10 15 Ser Leu Leu Glu 20 <210> 204 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 204 Lys Ala Phe Asp Asp lie Ala Lys Tyr Phe Ser Lys Glu Glu Trp Glu 1 5 10 15 Lys Met Lys Ala 20 <210> 205 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 205 Lys Ala Phe Asp Asp lie Ala Lys Tyr Phe Ser Lys Lys Glu Trp Glu 1 5 10 15 Lys Met Lys Ser 20 <210> 206 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 206 Leu Leu Gly Thr lie His Ala Leu lie Phe Ala Trp Asn Lys Trp lie 1 5 10 15 Asp lie Lys Gin 20 <210> 207 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 207 Phe Leu Pro lie Val Val Leu lie Phe Lys Ser lie Leu Phe Leu Pro 1 5 10 15 Cys Leu Arg Lys 20 <210> 208 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 208 Pro lie Lys lie Ala Ala lie lie Ala Ser Leu Thr Phe Leu Tyr Thr 1 5 10 15 Leu Leu Arg Glu 20 <210> 209 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 209 Lys lie Ala Ala lie lie Ala Ser Leu Thr Phe Leu Tyr Thr Leu Leu 1 5 10 15 Arg Glu Val lie 20 <210> 210 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 210 Leu Ala Leu Gly Leu Phe Val Cys Phe Tyr Ala Tyr Asn Phe Val Arg 1 5 10 15 Asp Val Leu Gln 20 <210> 211 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 211 Ala Val Thr Asp His Pro Asp Arg Leu Trp Ala Trp Glu Lys Phe Val 1 5 10 15 Tyr Leu Asp Glu 20 <210> 212 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 212 Leu Arg Ser Phe Phe Tyr Val Thr Glu Thr Thr Phe Gln Lys Asn Arg 1 5 10 15 Leu Phe Phe Tyr 20 <210> 213 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 213 Arg Ser Phe Phe Tyr Val Thr Glu Thr Thr Phe Gin Lys Asn Arg Leu 1 5 10 15 Phe Phe Tyr Arg 20 <210> 214 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 214 Leu Asp Ser Leu Gly Gin Trp Asn Tyr Thr Phe Ala Ser Thr Glu Gly 1 5 10 15 Gln Tyr Leu Leu 20 <210> 215 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 215 Leu Ser Asn Asn Ser Leu Val Ser Leu Thr Tyr Val Ser Phe Arg Asn 1 5 10 15 Leu Thr His Leu 20 <210> 216 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 216 Ala Pro Lys Ser Gin Phe Gly Arg He He His Thr Pro Phe Met Lys 1 5 10 15 Phe He He His 20 <210> 217 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 217 He He His Gly Ala Ser Tyr Phe Thr Phe Leu Leu Leu Leu Asn Leu 1 5 10 15 Tyr Ser Leu Val 20 <210> 218 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 218 Asn Gin Leu Ser Phe Val Met Asn Ser Leu Tyr Leu Ala Thr Phe Ala 1 5 10 15 Leu Lys Val Val 20 <210> 219 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 219 Gln Gln Ser Asn Asp Thr Phe His Ser Phe He Gly Thr Cys Phe Ala 1 5 10 15 Leu Phe Trp Tyr 20 <210> 220 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic Construct <400> 220 Pro Leu Leu Thr Thr Lys Arg Val Phe Trp Lys Gly Val Leu Glu Glu 1 5 10 15 Leu Leu Trp Phe 20 <210> 221 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic Construct <400> 221 Asp Pro He Phe Leu Leu His His Ala Phe Val Asp Ser He Phe Glu 1 5 10 15 Gln Trp Leu Arg 20 <210> 222 <211> 20 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic Construct <400> 222 Gln Arg Cys Leu Arg Asp lie Leu Asp Gly Phe Phe Pro Ser Glu Leu 1 5 10 15 Gln Arg Leu Tyr 20 <210> 223 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 223 Leu Ala Ala Ala Ser Gly Gin Asn Arg Met Thr Gin Gly Gin His Phe 1 5 10 15 Leu Gin Lys Val 20 <210> 224 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 224 Arg Arg Cys Asp Arg Val Thr Arg Leu Glu Phe Leu Pro Phe His Phe 1 5 10 15 Leu Leu Ala Thr 20 <210> 225 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 225 Cys Ala Thr Trp Lys Val lie Cys Lys Ser Cys lie Ser Gin Thr Pro 1 5 10 15 Gly lie Asn Leu 20 <210> 226 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 226 Leu Gin lie Gin Ser Leu lie Ser Cys Trp Ala Phe Trp Thr Thr Trp 1 5 10 15 Thr Gin Ser Cys 20 <210> 227 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 227 Val Ser Thr Ala Asn Val Gly Asp lie Leu Ser Ala Ala Arg Leu Leu 1 5 10 15 Glu lie Pro Ala 20 <210> 228 <211> 24 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 228 Leu Pro Lys Pro Pro Lys Pro Val Ser Lys Met Arg Met Ala Thr Pro 1 5 10 15 Leu Leu Met Gln Ala Leu Pro Met 20 <210> 229 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 229 Lys Lys Leu Leu Thr Gin His Phe Val Gin Glu Asn Tyr Leu Glu Tyr 1 5 10 15 <210> 230 <211> 21 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 230 Pro Gly Cys Asn Lys Arg Tyr Phe Lys Leu Ser His Leu Gin Met His 1 5 10 15 Ser Arg Lys His Thr 20 <210> 231 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 231 Ser Leu Leu Met Trp He Thr Gin Cys Phe Leu Pro Val Phe 1 5 10 <210> 232 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 232 Tyr Leu Ala Met Pro Phe Ala Thr Pro Met Glu Ala Glu Leu Ala Arg 1 5 10 15 Arg Ser Leu Ala 20 <210> 233 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 233 Pro Lys Tyr Val Lys Gin Asn Thr Leu Lys Leu Ala Thr 1 5 10 <210> 234 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 234 Phe Arg Asp Tyr Val Asp Arg Phe Tyr Lys Thr Leu Arg Ala Glu Gin 1 5 10 15 Ala Ser Gin Glu 20 <210> 235 <211> 20 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 235 Thr Gly Ser Asn Cys Pro Pro His lie Glu Asn Phe Ser Asp lie Asp 1 5 10 15 Met Gly Glu lie 20 <210> 236 <211> 12 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 236 Glu Thr Leu Leu Arg Ala Val Glu Ser Tyr Leu Leu 1 5 10 <210> 237 <211> 16 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 237 Arg Gly Tyr Phe Lys Met Arg Thr Gly Lys Ser Ser lie Met Arg Ser 1 5 10 15 <210> 238 <211> 282 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 238 Met Met Arg Pro lie Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala Arg Ala Thr Pro Glu Asn Tyr Leu Phe Gin Gly Arg Gin Glu Cys 20 25 30 Tyr Ala Phe Asn Gly Thr Gin Arg Phe Leu Glu Arg Tyr He Tyr Asn 35 40 45 Arg Glu Glu Phe Ala Arg Phe Asp Ser Asp Val Gly Glu Phe Arg Ala 50 55 60 Val Thr Glu Leu Gly Arg Pro Ala Ala Glu Tyr Trp Asn Ser Gin Lys 65 70 75 80 Asp He Leu Glu Glu Lys Arg Ala Val Pro Asp Arg Met Cys Arg His 85 90 95 Asn Tyr Glu Leu Gly Gly Pro Met Thr Leu Gin Arg Arg Val Gin Pro 100 105 110 Arg Val Asn Val Ser Pro Ser Lys Lys Gly Pro Leu Gin His His Asn 115 120 125 Leu Leu Val Cys His Val Thr Asp Phe Tyr Pro Gly Ser He Gin Val 130 135 140 Arg Trp Phe Leu Asn Gly Gin Glu Glu Thr Ala Gly Val Val Ser Thr 145 150 155 160 Asn Leu He Arg Asn Gly Asp Trp Thr Phe Gin He Leu Val Met Leu 165 170 175 Glu Met Thr Pro Gin Gin Gly Asp Val Tyr Thr Cys Gin Val Glu His 180 185 190 Thr Ser Leu Asp Ser Pro Val Thr Val Glu Trp Lys Ala Gin Ser Asp 195 200 205 Ser Ala Arg Ser Lys Gly Gly Gly Gly Ser Leu Glu lie Glu Ala Ala 210 215 220 Phe Leu Glu Arg Glu Asn Thr Ala Leu Glu Thr Arg Val Ala Glu Leu 225 230 235 240 Arg Gin Arg Val Gin Arg Leu Arg Asn Arg Val Ser Gin Tyr Arg Thr 245 250 255 Arg Tyr Gly Pro Leu Gly Gly Gly Lys Gly Ser Gly Leu Asn Asp lie 260 265 270 Phe Glu Ala Gin Lys lie Glu Trp His Glu 275 280 <210> 239 <211> 849 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 239 Ala Thr Gly Ala Thr Gly Ala Gly Gly Cys Cys Cys Ala Thr Cys Gly 1 5 10 15 Thr Gly Cys Thr Gly Gly Thr Gly Cys Thr Gly Cys Thr Gly Thr Thr 20 25 30 Cys Gly Cys Cys Ala Cys Ala Thr Cys Thr Gly Cys Cys Cys Thr Gly 35 40 45 Gly Cys Cys Ala Gly Ala Gly Cys Cys Ala Cys Cys Cys Cys Cys Gly 50 55 60 Ala Gly Ala Ala Cys Thr Ala Cys Cys Thr Gly Thr Thr Thr Cys Ala 65 70 75 80 Gly Gly Gly Cys Cys Gly Gly Cys Ala Gly Gly Ala Ala Thr Gly Cys 85 90 95 Thr Ala Cys Gly Cys Cys Thr Thr Cys Ala Ala Cys Gly Gly Cys Ala 100 105 110 Cys Cys Cys Ala Gly Cys Gly Gly Thr Thr Thr Cys Thr Gly Gly Ala 115 120 125 Ala Cys Gly Gly Thr Ala Cys Ala Thr Cys Thr Ala Cys Ala Ala Cys 130 135 140 Cys Gly Gly Gly Ala Ala Gly Ala Gly Thr Thr Cys Gly Cys Cys Ala 145 150 155 160 Gly Ala Thr Thr Cys Gly Ala Cys Ala Gly Cys Gly Ala Cys Gly Thr 165 170 175 Gly Gly Gly Cys Gly Ala Gly Thr Thr Cys Ala Gly Ala Gly Cys Cys 180 185 190 Gly Thr Gly Ala Cys Ala Gly Ala Gly Cys Thr Gly Gly Gly Cys Ala 195 200 205 Gly Ala Cys Cys Thr Gly Cys Cys Gly Cys Cys Gly Ala Gly Thr Ala 210 215 220 Cys Thr Gly Gly Ala Ala Cys Ala Gly Cys Cys Ala Gly Ala Ala Gly 225 230 235 240 Gly Ala Cys Ala Thr Cys Cys Thr Gly Gly Ala Ala Gly Ala Gly Ala 245 250 255 Ala Gly Cys Gly Gly Gly Cys Cys Gly Thr Gly Cys Cys Cys Gly Ala 260 265 270 Cys Cys Gly Gly Ala Thr Gly Thr Gly Cys Ala Gly Ala Cys Ala Cys 275 280 285 Ala Ala Thr Thr Ala Cys Gly Ala Gly Cys Thr Gly Gly Gly Ala Gly 290 295 300 Gly Cys Cys Cys Cys Ala Thr Gly Ala Cys Cys Cys Thr Gly Cys Ala 305 310 315 320 Gly Ala Gly Ala Ala Gly Ala Gly Thr Gly Cys Ala Gly Cys Cys Cys 325 330 335​​​​​​​​​​ Ala Gly Ala Gly Thr Gly Ala Ala Cys Gly Thr Gly Thr Cys Cys Cys 340 345 350 Cys Cys Ala Gly Cys Ala Ala Gly Ala Ala Gly Gly Gly Cys Cys Cys 355 360 365 Cys Cys Thr Gly Cys Ala Gly Cys Ala Cys Cys Ala Cys Ala Ala Cys 370 375 380 Thr Thr Gly Cys Thr Thr Gly Thr Cys Thr Gly Cys Cys Ala Cys Gly 385 390 395 400 Thr Gly Ala Cys Cys Gly Ala Cys Thr Thr Cys Thr Ala Cys Cys Cys 405 410 415 Cys Gly Gly Cys Thr Cys Thr Ala Thr Cys Cys Ala Ala Gly Thr Gly 420 425 430 Cys Gly Gly Thr Gly Gly Thr Thr Cys Cys Thr Gly Ala Ala Cys Gly 435 440 445 Gly Cys Cys Ala Gly Gly Ala Ala Gly Ala Gly Ala Cys Ala Gly Cys 450 455 460 Cys Gly Gly Cys Gly Thr Gly Gly Thr Gly Thr Cys Cys Ala Cys Cys 465 470 475 480 Ala Ala Cys Cys Thr Gly Ala Thr Cys Ala Gly Ala Ala Ala Cys Gly 485 490 495 Gly Cys Gly Ala Cys Thr Gly Gly Ala Cys Cys Thr Thr Cys Cys Ala 500 505 510 Gly Ala Thr Cys Cys Thr Cys Gly Thr Gly Ala Thr Gly Cys Thr Gly 515 520 525 Gly Ala Ala Ala Thr Gly Ala Cys Cys Cys Cys Cys Cys Ala Gly Cys 530 535 540 Ala Gly Gly Gly Cys Gly Ala Cys Gly Thr Gly Thr Ala Cys Ala Cys 545 550 555 560 Cys Thr Gly Thr Cys Ala Gly Gly Thr Gly Gly Ala Ala Cys Ala Cys 565 570 575 Ala Cys Cys Ala Gly Cys Cys Thr Gly Gly Ala Cys Ala Gly Cys Cys 580 585 590 Cys Cys Gly Thr Gly Ala Cys Cys Gly Thr Gly Gly Ala Ala Thr Gly 595 600 605 Gly Ala Ala Gly Gly Cys Cys Cys Ala Gly Ala Gly Cys Gly Ala Thr 610 615 620 Ala Gly Cys Gly Cys Cys Ala Gly Ala Ala Gly Cys Ala Ala Ala Gly 625 630 635 640 Gly Cys Gly Gly Cys Gly Gly Ala Gly Gly Cys Ala Gly Cys Cys Thr 645 650 655 Gly Gly Ala Ala Ala Thr Cys Gly Ala Gly Gly Cys Cys Gly Cys Cys 660 665 670 Thr Thr Cys Cys Thr Gly Gly Ala Ala Ala Gly Ala Gly Ala Gly Ala 675 680 685 Ala Cys Ala Cys Cys Gly Cys Cys Cys Thr Gly Gly Ala Ala Ala Cys 690 695 700 Cys Cys Gly Gly Gly Thr Gly Gly Cys Cys Gly Ala Gly Cys Thr Gly 705 710 715 720 Ala Gly Ala Cys Ala Gly Ala Gly Ala Gly Thr Gly Cys Ala Gly Ala 725 730 735 Gly Ala Cys Thr Gly Cys Gly Gly Ala Ala Cys Cys Gly Gly Gly Thr 740 745 750 Gly Thr Cys Cys Cys Ala Gly Thr Ala Cys Cys Gly Gly Ala Cys Cys 755 760 765 Ala Gly Ala Thr Ala Thr Gly Gly Cys Cys Cys Thr Cys Thr Gly Gly 770 775 780 Gly Ala Gly Gly Cys Gly Gly Cys Ala Ala Ala Gly Gly Gly Thr Cys 785 790 795 800 Cys Gly Gly Cys Thr Thr Gly Ala Ala Cys Gly Ala Cys Ala Thr Thr 805 810 815 Thr Thr Thr Gly Ala Gly Gly Cys Cys Cys Ala Gly Ala Ala Gly Ala 820 825 830 Thr Ala Gly Ala Gly Thr Gly Gly Cys Ala Cys Gly Ala Gly Thr Gly 835 840 845 Ala <210> 240 <211> 282 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 240 Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala Arg Ala Thr Pro Glu Asn Tyr Leu Phe Gln Gly Arg Gln Glu Cys 20 25 30 Tyr Ala Phe Asn Gly Thr Gln Arg Phe Leu Glu Arg Tyr Ile Tyr Asn 35 40 45 Arg Glu Glu Phe Ala Arg Phe Asp Ser Asp Val Gly Glu Phe Arg Ala 50 55 60 Val Thr Glu Leu Gly Arg Pro Ala Ala Glu Tyr Trp Asn Ser Gln Lys 65 70 75 80 Asp Ile Leu Glu Glu Lys Arg Ala Val Pro Asp Arg Met Cys Arg His 85 90 95 Asn Tyr Glu Leu Gly Gly Pro Met Thr Leu Gln Arg Arg Val Gln Pro 100 105 110 Arg Val Asn Val Ser Pro Ser Lys Lys Gly Pro Leu Gln His His Asn 115 120 125 Trp Leu Val Cys His Val Thr Asp Phe Tyr Pro Gly Ser Ile Gln Val 130 135 140 Arg Trp Phe Leu Asn Gly Gln Glu Glu Thr Ala Gly Val Met Ser Thr 145 150 155 160 Asn Leu Ile Arg Asn Gly Asp Trp Thr Phe Gln Ile Leu Val Met Leu 165 170 175 Glu Met Thr Pro Gln Gln Gly Asp Val Tyr Thr Cys Gln Val Glu His 180 185 190 Thr Ser Leu Asp Ser Pro Val Thr Val Glu Trp Lys Ala Gln Ser Asp 195 200 205 Ser Ala Arg Ser Lys Gly Gly Gly Gly Ser Leu Glu Ile Glu Ala Ala 210 215 220 Phe Leu Glu Arg Glu Asn Thr Ala Leu Glu Thr Arg Val Ala Glu Leu 225 230 235 240 Arg Gln Arg Val Gln Arg Leu Arg Asn Arg Val Ser Gln Tyr Arg Thr 245 250 255 Arg Tyr Gly Pro Leu Gly Gly Gly Lys Gly Ser Gly Leu Asn Asp Ile 260 265 270 Phe Glu Ala Gln Lys Ile Glu Trp His Glu 275 280 <210> 241 <211> 849 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 241 Ala Thr Gly Ala Thr Gly Ala Gly Gly Cys Cys Cys Ala Thr Cys Gly 1 5 10 15 Thr Gly Cys Thr Gly Gly Thr Gly Cys Thr Gly Cys Thr Gly Thr Thr 20 25 30 Cys Gly Cys Cys Ala Cys Ala Thr Cys Thr Gly Cys Cys Cys Thr Gly 35 40 45 Gly Cys Cys Ala Gly Ala Gly Cys Cys Ala Cys Cys Cys Cys Cys Gly 50 55 60 Ala Gly Ala Ala Cys Thr Ala Cys Cys Thr Gly Thr Thr Thr Cys Ala 65 70 75 80 Gly Gly Gly Cys Cys Gly Gly Cys Ala Gly Gly Ala Ala Thr Gly Cys 85 90 95 Thr Ala Cys Gly Cys Cys Thr Thr Cys Ala Ala Cys Gly Gly Cys Ala 100 105 110 Cys Cys Cys Ala Gly Cys Gly Gly Thr Thr Thr Cys Thr Gly Gly Ala 115 120 125 Ala Cys Gly Gly Thr Ala Cys Ala Thr Cys Thr Ala Cys Ala Ala Cys 130 135 140 Cys Gly Gly Gly Ala Ala Gly Ala Gly Thr Thr Cys Gly Cys Cys Ala 145 150 155 160 Gly Ala Thr Thr Cys Gly Ala Cys Ala Gly Cys Gly Ala Cys Gly Thr 165 170 175 Gly Gly Gly Cys Gly Ala Gly Thr Thr Cys Ala Gly Ala Gly Cys Cys 180 185 190 Gly Thr Gly Ala Cys Ala Gly Ala Gly Cys Thr Gly Gly Gly Cys Ala 195 200 205 Gly Ala Cys Cys Thr Gly Cys Cys Gly Cys Cys Gly Ala Gly Thr Ala 210 215 220 Cys Thr Gly Gly Ala Ala Cys Ala Gly Cys Cys Ala Gly Ala Ala Gly 225 230 235 240 Gly Ala Cys Ala Thr Cys Cys Thr Gly Gly Ala Ala Gly Ala Gly Ala 245 250 255 Ala Gly Cys Gly Gly Gly Cys Cys Gly Thr Gly Cys Cys Cys Gly Ala 260 265 270 Cys Cys Gly Gly Ala Thr Gly Thr Gly Cys Ala Gly Ala Cys Ala Cys 275 280 285 Ala Ala Thr Thr Ala Cys Gly Ala Gly Cys Thr Gly Gly Gly Ala Gly 290 295 300 Gly Cys Cys Cys Cys Ala Thr Gly Ala Cys Cys Cys Thr Gly Cys Ala 305 310 315 320 Gly Ala Gly Ala Ala Gly Ala Gly Thr Gly Cys Ala Gly Cys Cys Cys 325 330 335 Ala Gly Ala Gly Thr Gly Ala Ala Cys Gly Thr Gly Thr Cys Cys Cys 340 345 350 Cys Cys Ala Gly Cys Ala Ala Gly Ala Ala Gly Gly Gly Cys Cys Cys 355 360 365 Cys Cys Thr Gly Cys Ala Gly Cys Ala Cys Cys Ala Cys Ala Ala Cys 370 375 380 Thr Gly Gly Cys Thr Thr Gly Thr Cys Thr Gly Cys Cys Ala Cys Gly 385 390 395 400 Thr Gly Ala Cys Cys Gly Ala Cys Thr Thr Cys Thr Ala Cys Cys Cys 405 410 415 Cys Gly Gly Cys Thr Cys Thr Ala Thr Cys Cys Ala Gly Thr Gly 420 425 430 Cys Gly Gly Thr Gly Gly Thr Thr Cys Cys Thr Gly Ala Ala Cys Gly 435 440 445 Gly Cys Cys Ala Gly Gly Ala Ala Gly Ala Gly Ala Cys Ala Gly Cys 450 455 460 Cys Gly Gly Cys Gly Thr Gly Ala Thr Gly Thr Cys Cys Ala Cys Cys 465 470 475 480 Ala Ala Cys Cys Thr Gly Ala Thr Cys Ala Gly Ala Ala Ala Cys Gly 485 490 495 Gly Cys Gly Ala Cys Thr Gly Gly Ala Cys Cys Thr Thr Cys Cys Ala 500 505 510 Gly Ala Thr Cys Cys Thr Cys Gly Thr Gly Ala Thr Gly Cys Thr Gly 515 520 525 Gly Ala Ala Ala Thr Gly Ala Cys Cys Cys Cys Cys Cys Ala Gly Cys 530 535 540 Ala Gly Gly Gly Cys Gly Ala Cys Gly Thr Gly Thr Ala Cys Ala Cys 545 550 555 560 Cys Thr Gly Thr Cys Ala Gly Gly Thr Gly Gly Ala Ala Cys Ala Cys 565 570 575 Ala Cys Cys Ala Gly Cys Cys Thr Gly Gly Ala Cys Ala Gly Cys Cys 580 585 590 Cys Cys Gly Thr Gly Ala Cys Cys Gly Thr Gly Gly Ala Ala Thr Gly 595 600 605 Gly Ala Ala Gly Gly Cys Cys Cys Ala Gly Ala Gly Cys Gly Ala Thr 610 615 620 Ala Gly Cys Gly Cys Cys Ala Gly Ala Ala Gly Cys Ala Ala Ala Gly 625 630 635 640 Gly Cys Gly Gly Cys Gly Gly Ala Gly Gly Cys Ala Gly Cys Cys Thr 645 650 655 Gly Gly Ala Ala Ala Thr Cys Gly Ala Gly Gly Cys Cys Gly Cys Cys 660 665 670 Thr Thr Cys Cys Thr Gly Gly Ala Ala Ala Gly Ala Gly Ala Gly Ala 675 680 685 Ala Cys Ala Cys Cys Gly Cys Cys Cys Thr Gly Gly Ala Ala Ala Cys 690 695 700 Cys Cys Gly Gly Gly Thr Gly Gly Cys Cys Gly Ala Gly Cys Thr Gly 705 710 715 720 Ala Gly Ala Cys Ala Gly Ala Gly Ala Gly Thr Gly Cys Ala Gly Ala 725 730 735 Gly Ala Cys Thr Gly Cys Gly Gly Ala Ala Cys Cys Gly Gly Gly Thr 740 745 750 Gly Thr Cys Cys Cys Ala Gly Thr Ala Cys Cys Gly Gly Ala Cys Cys 755 760 765 Ala Gly Ala Thr Ala Thr Gly Gly Cys Cys Cys Thr Cys Thr Gly Gly 770 775 780 Gly Ala Gly Gly Cys Gly Gly Cys Ala Ala Ala Gly Gly Gly Thr Cys 785 790 795 800 Cys Gly Gly Cys Thr Thr Gly Ala Ala Cys Gly Ala Cys Ala Thr Thr 805 810 815 Thr Thr Thr Gly Ala Gly Gly Cys Cys Cys Ala Gly Ala Ala Gly Ala 820 825 830 Thr Ala Gly Ala Gly Thr Gly Gly Cys Ala Cys Gly Ala Gly Thr Gly 835 840 845 Ala <210> 242 <211> 273 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 242 Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala Ile Lys Ala Asp His Val Ser Thr Tyr Ala Ala Phe Val Gln Thr 20 25 30 His Arg Pro Thr Gly Glu Phe Met Phe Glu Phe Asp Glu Asp Glu Met 35 40 45 Phe Tyr Val Asp Leu Asp Lys Lys Glu Thr Val Trp His Leu Glu Glu 50 55 60 Phe Gly Gln Ala Phe Ser Phe Glu Ala Gln Gly Gly Leu Ala Asn Ile 65 70 75 80 Ala Ile Leu Asn Asn Asn Leu Asn Thr Leu Ile Gln Arg Ser Asn His 85 90 95 Thr Gln Ala Thr Asn Asp Pro Pro Glu Val Thr Val Phe Pro Lys Glu 100 105 110 Pro Val Glu Leu Gly Gln Pro Asn Thr Leu Ile Cys His Ile Asp Lys 115 120 125 Phe Phe Pro Pro Val Leu Asn Val Thr Trp Leu Cys Asn Gly Glu Leu 130 135 140 Val Thr Glu Gly Val Ala Glu Ser Leu Phe Leu Pro Arg Thr Asp Tyr 145 150 155 160 Ser Phe His Lys Phe His Tyr Leu Thr Phe Val Pro Ser Ala Glu Asp 165 170 175 Phe Tyr Asp Cys Arg Val Glu His Trp Gly Leu Asp Gln Pro Leu Leu 180 185 190 Lys His Trp Glu Ala Gln Glu Pro Ile Gln Met Pro Glu Thr Thr Glu 195 200 205 Thr Gly Gly Gly Gly Ser Leu Glu Ile Arg Ala Ala Phe Leu Arg Gln 210 215 220 Arg Asn Thr Ala Leu Arg Thr Glu Val Ala Glu Leu Glu Gln Glu Val 225 230 235 240 Gln Arg Leu Glu Asn Glu Val Ser Gln Tyr Glu Thr Arg Tyr Gly Pro 245 250 255 Leu Gly Gly Gly Lys Gly Ser His His His His His His His His His 260 265 270 His <210> 243 <211> 822 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 243 Ala Thr Gly Ala Thr Gly Ala Gly Gly Cys Cys Cys Ala Thr Cys Gly 1 5 10 15 Thr Gly Cys Thr Gly Gly Thr Gly Cys Thr Gly Cys Thr Gly Thr Thr 20 25 30 Cys Gly Cys Cys Ala Cys Ala Thr Cys Thr Gly Cys Cys Cys Thr Gly 35 40 45 Gly Cys Cys Ala Thr Cys Ala Ala Gly Gly Cys Cys Gly Ala Cys Cys 50 55 60 Ala Cys Gly Thr Gly Thr Cys Cys Ala Cys Ala Thr Ala Cys Gly Cys 65 70 75 80 Cys Gly Cys Cys Thr Thr Cys Gly Thr Gly Cys Ala Gly Ala Cys Cys 85 90 95 Cys Ala Cys Ala Gly Ala Cys Cys Cys Ala Cys Cys Gly Gly Cys Gly 100 105 110 Ala Gly Thr Thr Cys Ala Thr Gly Thr Thr Cys Gly Ala Gly Thr Thr 115 120 125 Cys Gly Ala Cys Gly Ala Gly Gly Ala Cys Gly Ala Gly Ala Thr Gly 130 135 140 Thr Thr Cys Thr Ala Cys Gly Thr Gly Gly Ala Cys Cys Thr Gly Gly 145 150 155 160 Ala Cys Ala Ala Gly Ala Ala Ala Gly Ala Ala Ala Cys Cys Gly Thr 165 170 175 Gly Thr Gly Gly Cys Ala Cys Cys Thr Gly Gly Ala Ala Gly Ala Gly 180 185 190 Thr Thr Cys Gly Gly Cys Cys Ala Gly Gly Cys Cys Thr Thr Cys Ala 195 200 205 Gly Cys Thr Thr Thr Gly Ala Gly Gly Cys Cys Cys Ala Gly Gly Gly 210 215 220 Cys Gly Gly Ala Cys Thr Gly Gly Cys Cys Ala Ala Thr Ala Thr Cys 225 230 235 240 Gly Cys Cys Ala Thr Cys Cys Thr Gly Ala Ala Cys Ala Ala Cys Ala 245 250 255 Ala Cys Cys Thr Gly Ala Ala Cys Ala Cys Cys Cys Thr Gly Ala Thr 260 265 270 Cys Cys Ala Gly Cys Gly Gly Ala Gly Cys Ala Ala Cys Cys Ala Cys 275 280 285 Ala Cys Cys Cys Ala Gly Gly Cys Cys Ala Cys Cys Ala Ala Cys Gly 290 295 300 Ala Thr Cys Cys Cys Cys Cys Cys Gly Ala Ala Gly Thr Gly Ala Cys 305 310 315 320 Cys Gly Thr Gly Thr Thr Cys Cys Cys Cys Ala Ala Ala Gly Ala Ala 325 330 335 Cys Cys Cys Gly Thr Gly Gly Ala Ala Cys Thr Gly Gly Gly Cys Cys 340 345 350 Ala Gly Cys Cys Cys Ala Ala Thr Ala Cys Cys Cys Thr Gly Ala Thr 355 360 365 Cys Thr Gly Cys Cys Ala Cys Ala Thr Cys Gly Ala Cys Ala Ala Gly 370 375 380 Thr Thr Cys Thr Thr Cys Cys Cys Cys Cys Cys Cys Gly Thr Gly Cys 385 390 395 400 Thr Gly Ala Ala Cys Gly Thr Gly Ala Cys Cys Thr Gly Gly Cys Thr 405 410 415 Gly Thr Gly Cys Ala Ala Thr Gly Gly Cys Gly Ala Gly Cys Thr Cys 420 425 430 Gly Thr Gly Ala Cys Ala Gly Ala Gly Gly Gly Cys Gly Thr Gly Gly 435 440 445 Cys Cys Gly Ala Gly Thr Cys Thr Cys Thr Gly Thr Thr Cys Cys Thr 450 455 460 Gly Cys Cys Cys Ala Gly Ala Ala Cys Cys Gly Ala Cys Thr Ala Cys 465 470 475 480 Ala Gly Cys Thr Thr Cys Cys Ala Cys Ala Ala Gly Thr Thr Cys Cys 485 490 495 Ala Cys Thr Ala Cys Cys Thr Gly Ala Cys Cys Thr Thr Cys Gly Thr 500 505 510 Gly Cys Cys Cys Ala Gly Cys Gly Cys Cys Gly Ala Gly Gly Ala Cys 515 520 525 Thr Thr Cys Thr Ala Cys Gly Ala Cys Thr Gly Cys Ala Gly Ala Gly 530 535 540 Thr Gly Gly Ala Ala Cys Ala Cys Thr Gly Gly Gly Gly Cys Cys Thr 545 550 555 560 Gly Gly Ala Cys Cys Ala Gly Cys Cys Cys Cys Thr Gly Cys Thr Gly 565 570 575 Ala Ala Ala Cys Ala Thr Thr Gly Gly Gly Ala Ala Gly Cys Cys Cys 580 585 590 Ala Gly Gly Ala Ala Cys Cys Cys Ala Thr Cys Cys Ala Gly Ala Thr 595 600 605 Gly Cys Cys Cys Gly Ala Gly Ala Cys Ala Ala Cys Cys Gly Ala Gly 610 615 620 Ala Cys Ala Gly Gly Cys Gly Gly Cys Gly Gly Ala Gly Gly Cys Ala 625 630 635 640 Gly Cys Cys Thr Gly Gly Ala Ala Ala Thr Cys Ala Gly Ala Gly Cys 645 650 655 Cys Gly Cys Cys Thr Thr Cys Cys Thr Gly Cys Gly Gly Cys Ala Gly 660 665 670 Ala Gly Ala Ala Ala Cys Ala Cys Cys Gly Cys Cys Cys Thr Gly Ala 675 680 685 Gly Ala Ala Cys Cys Gly Ala Ala Gly Thr Gly Gly Cys Cys Gly Ala 690 695 700 Gly Cys Thr Gly Gly Ala Ala Cys Ala Gly Gly Ala Ala Gly Thr Gly 705 710 715 720 Cys Ala Gly Cys Gly Gly Cys Thr Gly Gly Ala Ala Ala Ala Cys Gly 725 730 735 Ala Gly Gly Thr Gly Thr Cys Cys Cys Ala Gly Thr Ala Cys Gly Ala 740 745 750 Gly Ala Cys Ala Ala Gly Ala Thr Ala Cys Gly Gly Cys Cys Cys Thr 755 760 765 Cys Thr Gly Gly Gly Ala Gly Gly Cys Gly Gly Cys Ala Ala Gly Gly 770 775 780 Gly Cys Thr Cys Thr Cys Ala Cys Cys Ala Cys Cys Ala Cys Cys Ala 785 790 795 800 Thr Cys Ala Cys Cys Ala Thr Cys Ala Thr Cys Ala Thr Cys Ala Cys 805 810 815 Cys Ala Thr Thr Gly Ala 820 <210> 244 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 244 Gly Leu Asn Asp He Phe Glu Ala Gin Lys He Glu Trp His Glu 1 5 10 15 <210> 245 <211> 1185 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 245 Ala Thr Gly Ala Thr Gly Cys Gly Gly Cys Cys Cys Ala Thr Cys Gly 1 5 10 15 Thr Gly Cys Thr Gly Gly Thr Gly Cys Thr Gly Cys Thr Gly Thr Thr 20 25 30 Thr Gly Cys Cys Ala Cys Ala Thr Cys Thr Gly Cys Cys Cys Thr Gly 35 40 45 Gly Cys Cys Ala Ala Gly Ala Ala Ala Gly Thr Gly Gly Thr Gly Cys 50 55 60 Thr Gly Gly Gly Cys Ala Ala Ala Ala Ala Ala Gly Gly Gly Gly Ala 65 70 75 80 Thr Ala Cys Ala Gly Thr Gly Gly Ala Ala Cys Thr Gly Ala Cys Cys 85 90 95 Thr Gly Thr Ala Cys Ala Gly Cys Thr Thr Cys Cys Cys Ala Gly Ala 100 105 110 Ala Gly Ala Ala Gly Ala Gly Cys Ala Thr Ala Cys Ala Ala Thr Thr 115 120 125 Cys Cys Ala Cys Thr Gly Gly Ala Ala Ala Ala Ala Cys Thr Cys Cys 130 135 140 Ala Ala Cys Cys Ala Gly Ala Thr Ala Ala Ala Gly Ala Thr Thr Cys 145 150 155 160 Thr Gly Gly Gly Ala Ala Ala Thr Cys Ala Gly Gly Gly Cys Thr Cys 165 170 175 Cys Thr Thr Cys Thr Thr Ala Ala Cys Thr Ala Ala Ala Gly Gly Thr 180 185 190 Cys Cys Ala Thr Cys Cys Ala Ala Gly Cys Thr Gly Ala Ala Thr Gly 195 200 205 Ala Thr Cys Gly Cys Gly Cys Thr Gly Ala Cys Thr Cys Ala Ala Gly 210 215 220 Ala Ala Gly Ala Ala Gly Cys Cys Thr Thr Thr Gly Gly Gly Ala Cys 225 230 235 240 Cys Ala Ala Gly Gly Ala Ala Ala Cys Thr Thr Thr Cys Cys Cys Cys 245 250 255 Thr Gly Ala Thr Cys Ala Thr Cys Ala Ala Gly Ala Ala Thr Cys Thr 260 265 270 Thr Ala Ala Gly Ala Thr Ala Gly Ala Ala Gly Ala Cys Thr Cys Ala 275 280 285 Gly Ala Thr Ala Cys Thr Thr Ala Cys Ala Thr Cys Thr Gly Thr Gly 290 295 300 Ala Ala Gly Thr Gly Gly Ala Gly Gly Ala Cys Cys Ala Gly Ala Ala 305 310 315 320 Gly Gly Ala Gly Gly Ala Gly Gly Thr Gly Cys Ala Ala Thr Thr Gly 325 330 335 Cys Thr Ala Gly Thr Gly Thr Thr Cys Gly Gly Ala Thr Thr Gly Ala 340 345 350 Cys Thr Gly Cys Cys Ala Ala Cys Thr Cys Thr Gly Ala Cys Ala Cys 355 360 365 Cys Cys Ala Cys Cys Thr Gly Cys Thr Thr Cys Ala Gly Gly Gly Gly 370 375 380 Cys Ala Gly Ala Gly Cys Cys Thr Gly Ala Cys Cys Cys Thr Gly Ala 385 390 395 400 Cys Cys Thr Thr Gly Gly Ala Gly Ala Gly Cys Cys Cys Cys Cys Cys 405 410 415 Thr Gly Gly Thr Ala Gly Thr Ala Gly Cys Cys Cys Cys Thr Cys Ala 420 425 430 Gly Thr Gly Cys Ala Ala Thr Gly Thr Ala Gly Gly Ala Gly Thr Cys 435 440 445 Cys Ala Ala Gly Gly Gly Gly Thr Ala Ala Ala Ala Ala Cys Ala Thr 450 455 460 Ala Cys Ala Gly Gly Gly Gly Gly Gly Gly Ala Ala Gly Ala Cys Cys 465 470 475 480 Cys Thr Cys Thr Cys Cys Gly Thr Gly Thr Cys Thr Cys Ala Gly Cys 485 490 495 Thr Gly Gly Ala Gly Cys Thr Cys Cys Ala Gly Gly Ala Thr Ala Gly 500 505 510 Thr Gly Gly Cys Ala Cys Cys Thr Gly Gly Ala Cys Ala Thr Gly Cys 515 520 525 Ala Cys Thr Gly Thr Cys Thr Thr Gly Cys Ala Gly Ala Ala Cys Cys 530 535 540 Ala Gly Ala Ala Gly Ala Ala Gly Gly Thr Gly Gly Ala Gly Thr Thr 545 550 555 560 Cys Ala Ala Ala Ala Thr Ala Gly Ala Cys Ala Thr Cys Gly Thr Gly 565 570 575 Gly Thr Gly Cys Thr Ala Gly Cys Thr Thr Thr Cys Cys Ala Gly Ala 580 585 590 Ala Gly Gly Cys Cys Thr Cys Cys Ala Gly Cys Ala Thr Ala Gly Thr 595 600 605 Cys Thr Ala Thr Ala Ala Gly Ala Ala Ala Gly Ala Gly Gly Gly Gly 610 615 620 Gly Ala Ala Cys Ala Gly Gly Thr Gly Gly Ala Gly Thr Thr Cys Thr 625 630 635 640 Cys Cys Thr Thr Cys Cys Cys Ala Cys Thr Cys Gly Cys Cys Thr Thr 645 650 655 Thr Ala Cys Ala Gly Thr Thr Gly Ala Ala Ala Ala Gly Cys Thr Gly 660 665 670 Ala Cys Gly Gly Gly Cys Ala Gly Thr Gly Gly Cys Gly Ala Gly Cys 675 680 685 Thr Gly Thr Gly Gly Thr Gly Gly Cys Ala Gly Gly Cys Gly Gly Ala 690 695 700 Gly Ala Gly Gly Gly Cys Thr Thr Cys Cys Thr Cys Cys Thr Cys Cys 705 710 715 720 Ala Ala Gly Thr Cys Thr Thr Gly Gly Ala Thr Cys Ala Cys Cys Thr 725 730 735 Thr Thr Gly Ala Cys Cys Thr Gly Ala Ala Gly Ala Ala Cys Ala Ala 740 745 750 Gly Gly Ala Ala Gly Thr Gly Thr Cys Thr Gly Thr Ala Ala Ala Ala 755 760 765 Cys Gly Gly Gly Thr Thr Ala Cys Cys Cys Ala Gly Gly Ala Cys Cys 770 775 780 Cys Thr Ala Ala Gly Cys Thr Cys Cys Ala Gly Ala Thr Gly Gly Gly 785 790 795 800 Cys Ala Ala Gly Ala Ala Gly Cys Thr Cys Cys Cys Gly Cys Thr Cys 805 810 815 Cys Ala Cys Cys Thr Cys Ala Cys Cys Cys Thr Gly Cys Cys Cys Cys 820 825 830 Ala Gly Gly Cys Cys Thr Thr Gly Cys Cys Thr Cys Ala Gly Thr Ala 835 840 845 Thr Gly Cys Thr Gly Gly Cys Thr Cys Thr Gly Gly Ala Ala Ala Cys 850 855 860 Cys Thr Cys Ala Cys Cys Cys Thr Gly Gly Cys Cys Cys Thr Thr Gly 865 870 875 880 Ala Ala Gly Cys Gly Ala Ala Ala Ala Cys Ala Gly Gly Ala Ala Ala 885 890 895 Gly Thr Thr Gly Cys Ala Thr Cys Ala Gly Gly Ala Ala Gly Thr Gly 900 905 910 Ala Ala Cys Cys Thr Gly Gly Thr Gly Gly Thr Gly Ala Thr Gly Ala 915 920 925 Gly Ala Gly Cys Cys Ala Cys Thr Cys Ala Gly Cys Thr Cys Cys Ala 930 935 940 Gly Ala Ala Ala Ala Ala Thr Thr Thr Gly Ala Cys Cys Thr Gly Thr 945 950 955 960 Gly Ala Gly Gly Thr Gly Thr Gly Gly Gly Gly Ala Cys Cys Cys Ala 965 970 975 Cys Cys Thr Cys Cys Cys Cys Thr Ala Ala Gly Cys Thr Gly Ala Thr 980 985 990 Gly Cys Thr Gly Ala Gly Cys Thr Thr Gly Ala Ala Ala Cys Thr Gly 995 1000 1005 Gly Ala Gly Ala Ala Cys Ala Ala Gly Gly Ala Gly Gly Cys Ala 1010 1015 1020 Ala Ala Gly Gly Thr Cys Thr Cys Gly Ala Ala Gly Cys Gly Gly 1025 1030 1035 Gly Ala Gly Ala Ala Gly Gly Cys Gly Gly Thr Gly Thr Gly Gly 1040 1045 1050 Gly Thr Gly Cys Thr Gly Ala Ala Cys Cys Cys Thr Gly Ala Gly 1055 1060 1065 Gly Cys Gly Gly Gly Gly Ala Thr Gly Thr Gly Gly Cys Ala Gly 1070 1075 1080 Thr Gly Thr Cys Thr Gly Cys Thr Gly Ala Gly Thr Gly Ala Cys 1085 1090 1095 Thr Cys Gly Gly Gly Ala Cys Ala Gly Gly Thr Cys Cys Thr Gly 1100 1105 1110 Cys Thr Gly Gly Ala Ala Thr Cys Cys Ala Ala Cys Ala Thr Cys 1115 1120 1125 Ala Ala Gly Gly Thr Thr Cys Thr Gly Cys Cys Cys Ala Cys Ala 1130 1135 1140 Thr Gly Gly Gly Gly Cys Ala Gly Cys Cys Ala Cys Cys Ala Cys 1145 1150 1155 Cys Ala Cys Cys Ala Thr Cys Ala Cys Cys Ala Thr Cys Ala Thr 1160 1165 1170 Cys Ala Thr Cys Ala Cys Cys Ala Thr Thr Gly Ala 1175 1180 1185 <210> 246 <211> 394 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 246 Met Met Arg Pro Ile Val Leu Val Leu Leu Phe Ala Thr Ser Ala Leu 1 5 10 15 Ala Lys Lys Val Val Leu Gly Lys Lys Gly Asp Thr Val Glu Leu Thr 20 25 30 Cys Thr Ala Ser Gln Lys Lys Ser Ile Gln Phe His Trp Lys Asn Ser 35 40 45 Asn Gln Ile Lys Ile Leu Gly Asn Gln Gly Ser Phe Leu Thr Lys Gly 50 55 60 Pro Ser Lys Leu Asn Asp Arg Ala Asp Ser Arg Arg Ser Leu Trp Asp 65 70 75 80 Gln Gly Asn Phe Pro Leu Ile Ile Lys Asn Leu Lys Ile Glu Asp Ser 85 90 95 Asp Thr Tyr Ile Cys Glu Val Glu Asp Gln Lys Glu Glu Val Gln Leu 100 105 110 Leu Val Phe Gly Leu Thr Ala Asn Ser Asp Thr His Leu Leu Gln Gly 115 120 125 Gln Ser Leu Thr Leu Thr Leu Glu Ser Pro Pro Gly Ser Ser Pro Ser 130 135 140 Val Gln Cys Arg Ser Pro Arg Gly Lys Asn Ile Gln Gly Gly Lys Thr 145 150 155 160 Leu Ser Val Ser Gln Leu Glu Leu Gln Asp Ser Gly Thr Trp Thr Cys 165 170 175 Thr Val Leu Gln Asn Gln Lys Lys Val Glu Phe Lys Ile Asp Ile Val 180 185 190 Val Leu Ala Phe Gln Lys Ala Ser Ser Ile Val Tyr Lys Lys Glu Gly 195 200 205 Glu Gln Val Glu Phe Ser Phe Pro Leu Ala Phe Thr Val Glu Lys Leu 210 215 220 Thr Gly Ser Gly Glu Leu Trp Trp Gln Ala Glu Arg Ala Ser Ser Ser 225 230 235 240 Lys Ser Trp lie Thr Phe Asp Leu Lys Asn Lys Glu Val Ser Val Lys 245 250 255 Arg Val Thr Gin Asp Pro Lys Leu Gin Met Gly Lys Lys Leu Pro Leu 260 265 270 His Leu Thr Leu Pro Gin Ala Leu Pro Gin Tyr Ala Gly Ser Gly Asn 275 280 285 Leu Thr Leu Ala Leu Glu Ala Lys Thr Gly Lys Leu His Gin Glu Val 290 295 300 Asn Leu Val Val Met Arg Ala Thr Gin Leu Gin Lys Asn Leu Thr Cys 305 310 315 320 Glu Val Trp Gly Pro Thr Ser Pro Lys Leu Met Leu Ser Leu Lys Leu 325 330 335 Glu Asn Lys Glu Ala Lys Val Ser Lys Arg Glu Lys Ala Val Trp Val 340 345 350 Leu Asn Pro Glu Ala Gly Met Trp Gin Cys Leu Leu Ser Asp Ser Gly 355 360 365 Gln Val Leu Leu Glu Ser Asn lie Lys Val Leu Pro Thr Trp Gly Ser 370 375 380 His His His His His His His His His His 385 390 <210> 247 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 247 Cys Val Val Ser Gly Gly Val Asn Gly Gly Ala Thr Asn Lys Leu Ile 1 5 10 15 Phe <210> 248 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 248 Cys Ala Ser Ser Leu Thr Gly Gly Val Ser Tyr Glu Gin Tyr Phe 1 5 10 15 <210> 249 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 249 Cys Ala Ala Ser Val Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe 1 5 10 15 <210> 250 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 250 Cys Ala Ser Ser Leu Gly Thr Gly Gly Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 251 <211> 16 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 251 Cys Ala Val Ser Gly Gly Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe 1 5 10 15 <210> 252 <211> 15 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 252 Cys Ser Val Gin Gly Gly Leu Asp Ser Asn Tyr Gly Tyr Thr Phe 1 5 10 15 <210> 253 <211> 15 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 253 Cys Ala Ala Ser Val Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe 1 5 10 15 <210> 254 <211> 15 <212> PRT <213> Artificial Sequence (Artificial Sequence)​​​​ <220> <223> synthetic construct <400> 254 Cys Ala Ser Ser Leu Gly Thr Gly Gly Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 255 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 255 Cys Ala Ala Ser Val Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe 1 5 10 15 <210> 256 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 256 Cys Ala Ser Ser Leu Gly Thr Gly Gly Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 257 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 257 Cys Ala Tyr Arg Ser Asn Asn Phe Asn Lys Phe Tyr Phe 1 5 10 <210> 258 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 258 Cys Ala Ser Ser Leu Asn Thr Gly Ala Gly Tyr Gly Tyr Thr Phe 1 5 10 15 <210> 259 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 259 Cys Ala Ala Ser Val Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe 1 5 10 15 <210> 260 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 260 Cys Ala Ser Ser Leu Gly Thr Gly Gly Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 261 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 261 Cys Ala Ala Ser Val Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe 1 5 10 15 <210> 262 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 262 Cys Ala Ser Ser Leu Gly Thr Gly Gly Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 263 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 263 Cys Ala Val Glu Glu Arg Thr Gly Gly Phe Lys Thr Ile Phe 1 5 10 <210> 264 <211> 18 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 264 Cys Ala Ser Ser Leu Pro Ser Gly Gly Ala Pro Gly Thr Gly Glu Leu 1 5 10 15 Phe Phe <210> 265 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 265 Cys Ala Val Ser Gly Gly Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe 1 5 10 15 <210> 266 <211> 15 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 266 Cys Ser Val Gln Gly Gly Leu Asp Ser Asn Tyr Gly Tyr Thr Phe 1 5 10 15 <210> 267 <211> 15 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 267 Cys Ala Ala Ser Val Arg Gly Ser Thr Leu Gly Arg Leu Tyr Phe 1 5 10 15 <210> 268 <211> 15 <212> PRT <213> Artificial Sequence (Artificial Sequence) <220> <223> Synthetic construct <400> 268 Cys Ala Ser Ser Leu Gly Thr Gly Gly Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 269 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 269 Cys Leu Val Gly Asp Leu Gly Ala Asn Ala Gly Asn Met Leu Thr Phe 1 5 10 15 <210> 270 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 270 Cys Ala Ser Ser Ile Ala Thr Thr Asn Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 271 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 271 Cys Leu Val Gly Asp Leu Gly Ala Asn Ala Gly Asn Met Leu Thr Phe 1 5 10 15 <210> 272 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 272 Cys Ala Ser Ser Leu Glu Thr Gly Thr Asn Tyr Glu Gln Tyr Phe 1 5 10 15 <210> 273 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 273 Cys Ala Val Ala Leu Tyr Gly Gly Ala Thr Asn Lys Leu Ile Phe 1 5 10 15 <210> 274 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 274 Cys Ala Ser Ser Leu Asp Ile Gly Asn Asn Glu Gin Phe Phe 1 5 10 <210> 275 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 275 Cys Ala Gly Arg Ser Gly Gly Ser Asn Tyr Lys Leu Thr Phe 1 5 10 <210> 276 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 276 Cys Ala Ser Ser lie Ala Thr Thr Asn Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 277 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 277 Cys Ala Gly Arg Ser Gly Gly Ser Asn Tyr Lys Leu Thr Phe 1 5 10 <210> 278 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 278 Cys Ala Ser Ser lie Ala Thr Thr Asn Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 279 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct <400> 279 Cys Ala Gly Arg Ser Gly Gly Ser Asn Tyr Lys Leu Thr Phe 1 5 10 <210> 280 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 280 Cys Ala Ser Ser lie Ala Thr Thr Asn Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 281 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 281 Cys Ala Gly Arg Ser Gly Gly Ser Asn Tyr Lys Leu Thr Phe 1 5 10 <210> 282 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 282 Cys Ala Ser Ser lie Ala Thr Thr Asn Thr Gly Glu Leu Phe Phe 1 5 10 15 <210> 283 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> synthetic construct <400> 283 Cys Ala Gly Arg Ser Gly Gly Ser Asn Tyr Lys Leu Thr Phe 1 5 10 <210> 284 <211> 15 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Construct <400> 284 Cys Ala Ser Ser lie Ala Thr Thr Asn Thr Gly Glu Leu Phe Phe 1 5 10 15

Claims

1. An HLA class II molecule comprising a DP β chain, wherein the DP β chain comprises an extracellular domain, wherein the amino acid sequence of the extracellular domain is shown in SEQ ID NO:

3.

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

4.

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

4. The HLA class II molecule as claimed in claim 3, wherein the amino acid sequence of the DP α chain is shown in SEQ ID NO:

6.

5. The HLA class II molecule of claim 3, wherein the amino acid sequence of the DP α chain is shown in SEQ ID NO:

8.

6. The HLA class II molecule as described in any one of claims 1 to 5, wherein it is the DP1 allele.

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

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

9. The HLA class II molecule according to any one of claims 1 to 8, wherein the DP β chain comprises a signal peptide.

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