Compositions and Methods for the Therapy and Diagnosis of Influenza

1 . A composition comprising:

(a) an isolated human antibody or antigen-binding fragment thereof that specifically binds to an epitope of the hemagglutinin (HA) glycoprotein of an influenza virus; and

(b) an isolated human monoclonal antibody or antigen-binding fragment thereof that specifically binds to an epitope in the extracellular domain of the matrix 2 ectodomain (M2e) polypeptide of an influenza virus.

2 . The composition of claim 1 , wherein said isolated human monoclonal antibody or antigen-binding fragment thereof that specifically binds an epitope of the M2e polypeptide is selected from the group consisting of:

(a) an antibody or antigen-binding fragment thereof comprising a heavy chain variable (VH) domain and a light chain variable (VL) domain, wherein the VH domain and the VL domain each comprise three complementarity determining regions 1 to 3 (CDR1-3), and wherein each CDR comprises the following amino acid sequences:

VH CDR1: SEQ ID NOs: 72, 103, 179, 187, 203, 211, 228, 252, 260, 268, 284, 293, or 301;

VH CDR2: SEQ ID NOs: 74, 105, 180, 188, 204, 212, 229, 237, 253, 261, 269, 285, or 294;

VH CDR3 SEQ ID NOs: 76, 107, 181, 189, 197, 205, 213, 230, 238, 254, 262, 270, 286, or 295;

VL CDR1: SEQ ID NOs: 59, 92, 184, 192, 208, 192, 233, 241, 265, or 273;

VL CDR2: SEQ ID NOs: 61, 94, 185, 193, 209, 217, 226, 234, 258, 274, or 282; and

VL CDR3: SEQ ID NOs: 63, 96, 186, 194, 210, 218, 243, 259, 267, 275, 291, or 300; and

(b) an antibody or antigen-binding fragment thereof comprising a heavy chain variable (VH) domain and a light chain variable (VL) domain, wherein the VH domain and the VL domain each comprise three complementarity determining regions 1 to 3 (CDR1-3), and wherein each CDR comprises the following amino acid sequences:

VH CDR1: SEQ ID NOs: 109, 112, 182, 190, 206, 214, 239, 255, 263, 271, 287, 296, or 304;

VH CDR2: SEQ ID NOs: 110, 113, 183, 191, 207, 215, 232, 240, 256, 264, 272, 288, or 297;

VH CDR3 SEQ ID NOs: 76, 107, 181, 189, 197, 205, 213, 230, 238, 254, 262, 270, 286, or 295;

VL CDR1: SEQ ID NOs: 59, 92, 184, 192, 208, 192, 223, 241, 265, or 273;

VL CDR2: SEQ ID NOs: 61, 94, 185, 193, 209, 217, 226, 234, 258, 274, or 282; and

VL CDR3: SEQ ID NOs: 63, 96, 186, 194, 210, 218, 243, 259, 267, 275, 291, or 300.

3 . The composition of claim 1 , wherein said isolated human antibody that specifically binds an epitope of the HA glycoprotein comprises a heavy chain variable region (VH) domain and a light chain variable (VL) domain, wherein the VH domain and the VL domain each comprise three complementarity determining regions 1 to 3 (CDR1-3), and wherein each CDR comprises the following amino acid sequences:

VH CDR1: SEQ ID NOs: 247, 571, 586, 597, 603, 609, 615, 627, 633, 637, 643, 649, 658, 664, 670, 303, 251, 242, or 222;

VH CDR2: SEQ ID NOs: 248, 572, 587, 592, 598, 604, 610, 616, 628, 634, 638, 644, 650, 655, 659, 665, 671, 306, 249, 307, or 221;

VH CDR3: SEQ ID NOs: 568, 573, 588, 593, 599, 605, 611, 617, 629, 635, 639, 645, 651, 656, 660, 666, 672, 725, 246, 290, or 220;

VL CDR1: SEQ ID NOs: 569, 574, 577, 580, 583, 589, 594, 612, 618, 621, 624, 640, 646, 652, 661, 667, 285, 289, 245, 224, or 219;

VL CDR2: SEQ ID NOs: 570, 575, 578, 581, 584, 590, 595, 601, 607, 613, 619, 622, 625, 631, 653, 662, 668, 305, 223, or 231;

VL CDR3: SEQ ID NOs: 289, 576, 579, 582, 585, 591, 596, 602, 608, 614, 620, 623, 626, 632, 636, 642, 648, 654, 657, 663, 669, 308, 250, 227, or 280.

4 . The composition of claim 1 , wherein said epitope of the HA glycoprotein is GVTNKVNSIIDK (SEQ ID NO: 198), GVTNKVNSIINK (SEQ ID NO: 283), GVTNKENSIIDK (SEQ ID NO: 202), GVTNKVNRIIDK (SEQ ID NO: 201), GITNKVNSVIEK (SEQ ID NO: 281), GITNKENSVIEK (SEQ ID NO: 257), GITNKVNSIIDK (SEQ ID NO: 225), and KITSKVNNIVDK (SEQ ID NO: 216).

5 . The composition of claim 1 , wherein said epitope of the M2e polypeptide is a discontinuous epitope.

6 . The composition of claim 1 , wherein said epitope of the M2e polypeptide comprises,

a) the amino acid at positions 2, 5, and 6 of MSLLTEVETPTRNEWGCRCNDSSD (SEQ ID NO: 1); or

b) the amino acid at positions 2, 5, and 6 of SLLTEV (SEQ ID NO: 42).

7 . The composition of claim 1 comprising:

(a) an isolated human anti-HA antibody, or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) domain, wherein the VH domain comprises the following amino acid sequences: SEQ ID NOs 309, 313, 317, 321, 325, 329, 333, 337, 341, 345, 349, 353, 357, 361, 365, 369, 373, 377, 381, 385, 389, 393, 397, 401, 405, 409, 199, 417, 423, 429, 435, 441, 447, 453, 459, 465, 471, 477, 483, 489, 495, 501, 507, 513, 519, 525, 531, 537, 543, 550, 556, or 562, and a light chain variable (VL) domain, wherein the VL domain comprises the following amino acid sequences: SEQ ID NOs 310, 314, 318, 322, 326, 330, 334, 338, 342, 346, 350, 354, 358, 362, 366, 370, 374, 378, 382, 386, 390, 394, 398, 402, 406, 410, 414, 420, 426, 432, 438, 444, 450, 456, 462, 468, 474, 480, 486, 492, 498, 504, 510, 516, 522, 528, 534, 540, 547, 553, 559, or 565; and

(b) an isolated anti-matrix 2 ectodomain (M2e) antibody, or antigen-binding fragment thereof, comprising a heavy chain variable (VH) domain, wherein the VH domain comprises the following amino acid sequences: SEQ ID NOs 44, 277, 276, 50, 236, 235, 116, 120, 124, 128, 132, 136, 140, 144, 148, 152, 156, 160, 164, 168, 172, or 176, and a light chain variable (VL) domain, wherein the VL domain comprises the following amino acid sequences: SEQ ID NOs 46, 292, 52, 118, 122, 126, 130, 134, 138, 142, 146, 150, 154, 158, 162, 166, 170, or 178.

8 . A multivalent vaccine composition comprising the composition of claim 1 .

9 . A pharmaceutical composition comprising the composition of claim 1 and a pharmaceutical carrier.

10 . A method for stimulating an immune response in a subject, comprising administering to the subject the composition of claim 9 .

11 . A method for the treatment of an influenza virus infection in a subject in need thereof, comprising administering to said subject the composition of claim 9 .

12 . The method of claim 11 , wherein the subject has been exposed to an influenza virus.

13 . The method of claim 12 , wherein the subject has not be diagnosed with an influenza infection.

14 . A method for the prevention of an influenza virus infection in a subject in need thereof, comprising administering to said subject the vaccine of claim 8 , prior to exposure of said subject to an influenza virus.

15 . The method of claim 11 , wherein the method further comprises administering an anti-viral drug, a viral entry inhibitor or a viral attachment inhibitor.

16 . The method of claim 15 , wherein said anti-viral drug is a neuraminidase inhibitor, a HA inhibitor, a sialic acid inhibitor or an M2 ion channel.

17 . The method of claim 16 , wherein said M2 ion channel inhibitor is amantadine or rimantadine.

18 . The method of claim 16 , wherein said neuraminidase inhibitor is zanamivir or oseltamivir phosphate.

19 . The method of claim 11 , further comprising administering a second anti-Influenza A antibody.

20 . The method of claim 19 , wherein said antibody is administered prior to or after exposure to Influenza virus.

21 . The method of claim 12 , wherein the subject is at risk of contracting an influenza infection.

22 . The method of claim 11 , wherein said composition is administered at a dose sufficient to promote viral clearance or eliminate influenza infected cells.

23 . A method for determining the presence of an Influenza virus infection in a subject, comprising the steps of:

(a) contacting a biological sample obtained from the subject with the antibody of claim 1 ;

(b) detecting an amount of the antibody that binds to the biological sample; and

(c) comparing the amount of antibody that binds to the biological sample to a control value, and therefrom determining the presence of the Influenza virus in the subject.

24 . The method of claim 23 , wherein the control value is determined by contacting a control sample obtained from the subject with the antibody according to claim 1 and detecting an amount of the antibody that binds to the control sample.