                         SEQUENCE LISTING

<110>  THE UNITED STATES OF AMERICA, as represented by the 
       Secretary, Department of Health and Human Services
       Perera, Liyanage P
       Waldmann, Thomas
 
<120>  CHIMERIC ANTIGEN RECEPTOR (CAR) THAT TARGETS CHEMOKINE RECEPTOR 
       CCR4 AND ITS USE

<130>  4239-97560-02

<150>  62/397,810
<151>  2016-09-21

<160>  19    

<170>  PatentIn version 3.5

<210>  1
<211>  118
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  VH of humanized affinity matured mAb1567

<400>  1

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
1               5                   10                  15      


Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ala Ser Ala 
            20                  25                  30          


Trp Met His Trp Met Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 
        35                  40                  45              


Gly Trp Ile Asn Pro Gly Asn Val Asn Thr Lys Tyr Asn Glu Lys Phe 
    50                  55                  60                  


Lys Gly Arg Ala Thr Leu Thr Val Asp Thr Ser Thr Asn Thr Ala Tyr 
65                  70                  75                  80  


Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                85                  90                  95      


Ala Arg Ser Thr Tyr Tyr Arg Pro Leu Asp Tyr Trp Gly Gln Gly Thr 
            100                 105                 110         


Leu Val Thr Val Ser Ser 
        115             


<210>  2
<211>  112
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  VL of humanized affinity matured mAb1567

<400>  2

Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 
1               5                   10                  15      


Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Ile Leu Tyr Ser 
            20                  25                  30          


Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 
        35                  40                  45              


Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 
    50                  55                  60                  


Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 
65                  70                  75                  80  


Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys His Gln 
                85                  90                  95      


Tyr Met Ser Ser Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 
            100                 105                 110         


<210>  3
<211>  17
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker

<400>  3

Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
1               5                   10                  15      


Ser 
    


<210>  4
<211>  22
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Signal protein

<400>  4

Met Asp Phe Gln Val Gln Ile Phe Ser Phe Leu Leu Ile Ser Ala Ser 
1               5                   10                  15      


Val Ile Met Ser Arg Gly 
            20          


<210>  5
<211>  22
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Signal Protein

<400>  5

Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro Ala Phe 
1               5                   10                  15      


Leu Leu Ile Pro Asp Thr 
            20          


<210>  6
<211>  236
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Immunoglobulin Domain

<400>  6

Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 
1               5                   10                  15      


Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 
            20                  25                  30          


Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 
        35                  40                  45              


Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 
    50                  55                  60                  


Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 
65                  70                  75                  80  


Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 
                85                  90                  95      


Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 
            100                 105                 110         


Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 
        115                 120                 125             


Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 
    130                 135                 140                 


Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 
145                 150                 155                 160 


Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 
                165                 170                 175     


Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 
            180                 185                 190         


Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 
        195                 200                 205             


Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 
    210                 215                 220                 


Ser Leu Ser Leu Ser Pro Gly Lys Lys Asp Pro Lys 
225                 230                 235     


<210>  7
<211>  93
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD8 Transmembrane Domain

<400>  7

Ala Leu Ser Asn Ser Ile Met Tyr Phe Ser His Phe Val Pro Val Phe 
1               5                   10                  15      


Leu Pro Ala Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro 
            20                  25                  30          


Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys 
        35                  40                  45              


Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala 
    50                  55                  60                  


Cys Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu 
65                  70                  75                  80  


Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Asn His Arg 
                85                  90              


<210>  8
<211>  42
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  4-1BB Signalling Domain

<400>  8

Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 
1               5                   10                  15      


Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 
            20                  25                  30          


Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 
        35                  40          


<210>  9
<211>  112
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD3 zeta domain

<400>  9

Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly 
1               5                   10                  15      


Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr 
            20                  25                  30          


Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys 
        35                  40                  45              


Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys 
    50                  55                  60                  


Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg 
65                  70                  75                  80  


Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala 
                85                  90                  95      


Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
            100                 105                 110         


<210>  10
<211>  516
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Chimeric Antigen Receptor

<400>  10

Met Asp Phe Gln Val Gln Ile Phe Ser Phe Leu Leu Ile Ser Ala Ser 
1               5                   10                  15      


Val Ile Met Ser Arg Gly Asp Ile Val Met Thr Gln Ser Pro Asp Ser 
            20                  25                  30          


Leu Ala Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser 
        35                  40                  45              


Gln Ser Ile Leu Tyr Ser Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr 
    50                  55                  60                  


Gln Gln Lys Pro Gly Gln Ser Pro Lys Leu Leu Ile Tyr Trp Ala Ser 
65                  70                  75                  80  


Thr Arg Glu Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly 
                85                  90                  95      


Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala 
            100                 105                 110         


Val Tyr Tyr Cys His Gln Tyr Met Ser Ser Tyr Thr Phe Gly Gln Gly 
        115                 120                 125             


Thr Lys Leu Glu Ile Lys Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 
    130                 135                 140                 


Gly Ser Gly Gly Gly Gly Ser Gln Val Gln Leu Val Gln Ser Gly Ala 
145                 150                 155                 160 


Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser 
                165                 170                 175     


Gly Tyr Thr Phe Ala Ser Ala Trp Met His Trp Met Arg Gln Ala Pro 
            180                 185                 190         


Gly Gln Gly Leu Glu Trp Ile Gly Trp Ile Asn Pro Gly Asn Val Asn 
        195                 200                 205             


Thr Lys Tyr Asn Glu Lys Phe Lys Gly Arg Ala Thr Leu Thr Val Asp 
    210                 215                 220                 


Thr Ser Thr Asn Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu 
225                 230                 235                 240 


Asp Thr Ala Val Tyr Tyr Cys Ala Arg Ser Thr Tyr Tyr Arg Pro Leu 
                245                 250                 255     


Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Leu Ser 
            260                 265                 270         


Asn Ser Ile Met Tyr Phe Ser His Phe Val Pro Val Phe Leu Pro Ala 
        275                 280                 285             


Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr 
    290                 295                 300                 


Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala 
305                 310                 315                 320 


Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile 
                325                 330                 335     


Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser 
            340                 345                 350         


Leu Val Ile Thr Leu Tyr Cys Asn His Arg Lys Arg Gly Arg Lys Lys 
        355                 360                 365             


Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr 
    370                 375                 380                 


Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly 
385                 390                 395                 400 


Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala 
                405                 410                 415     


Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg 
            420                 425                 430         


Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu 
        435                 440                 445             


Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn 
    450                 455                 460                 


Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met 
465                 470                 475                 480 


Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly 
                485                 490                 495     


Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala 
            500                 505                 510         


Leu Pro Pro Arg 
        515     


<210>  11
<211>  66
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA encoding signal sequence

<400>  11
atggactttc aagtgcagat ctttagtttc ctgctcataa gcgctagtgt gatcatgtcc       60

agagga                                                                  66


<210>  12
<211>  336
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA encoding VL of humanized affinity matured  mAb1567

<400>  12
gatattgtga tgactcaaag ccccgacagt ctggccgtgt ctttgggcga gagagccaca       60

atcaactgca agtcctcaca gagtatcctt tattcctcta atcagaagaa ttacctcgca      120

tggtatcaac aaaaacccgg acagagccct aagcttttga tctattgggc atctacccga      180

gaatcaggag tgccggaccg cttcagtgga tcaggatcag gcacagactt tacgctgaca      240

atatcctctc ttcaggccga agacgttgcc gtgtactact gccatcaata tatgtcaagc      300

tacacattcg gccagggcac caaactcgag attaag                                336


<210>  13
<211>  354
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA encoding VH of humanized affinity matured  mAb1567

<400>  13
caggttcagc tcgtgcaatc aggggcagag gtcaagaagc cgggtgcctc tgtgaaggtg       60

tcatgtaagg cctccgggta tacttttgcc agcgcctgga tgcattggat gaggcaggcg      120

cccggccagg gtctggagtg gattggttgg attaatcccg gaaacgtgaa tactaagtat      180

aacgagaagt ttaagggcag ggccacactc acagtcgaca caagcaccaa taccgcgtac      240

atggaacttt ccagcctccg gtccgaggac actgcggtgt attactgcgc acgctccacc      300

tattacagac cacttgatta ctggggccaa gggaccctgg tgaccgtgtc tagc            354


<210>  14
<211>  51
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA encoding a linker

<400>  14
tctagtggtg gcggaggcag tggcggagga ggctccgggg gcggagggtc c                51


<210>  15
<211>  279
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA encoding CD8 domains

<400>  15
gcactcagca attccatcat gtacttctct catttcgtgc cagtatttct gcctgccaag       60

ccaactacca cacctgcgcc acgccctccc acgcccgcac ccacaattgc ttcacagcct      120

ctttctctgc ggcctgaggc ttgtcgccca gcagccggag gcgccgtgca tacgcgcggc      180

cttgacttcg catgtgacat ctacatttgg gctcctttgg ctggaacctg cggggtgttg      240

ttgcttagtc tggtgattac cctctactgc aatcataga                             279


<210>  16
<211>  126
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA encoding 4-1BB signaling molecule

<400>  16
aagcgggggc gaaagaaact tctctatatc ttcaaacagc ctttcatgcg accagtgcag       60

acaacccaag aggaagacgg atgcagctgt cgctttccag aggaggaaga agggggctgc      120

gagctg                                                                 126


<210>  17
<211>  306
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA encoding a CD3 zeta domain

<400>  17
agagtgaaat tctctcgctc cgctgacgcc cccgcgtatc aacagggcca gaatcagctc       60

tacaacgaac ttaaccttgg gcggagagaa gaatacgatg ttctcgacaa gcgcaggggg      120

agagaccctg agatgggcgg gaaaccgcgc cgcaagaacc cccaagaagg gttgtataac      180

gagctccaga aggacaaaat ggctgaagcc tactcagaga taggtatgaa gggcgagcgc      240

cgcagaggga agggacacga tggtctgtac caaggccttt caaccgccac caaggatacc      300

tatgat                                                                 306


<210>  18
<211>  1565
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA encoding a chimeric antigen receptor

<400>  18
gcggccgcat ggactttcaa gtgcagatct ttagtttcct gctcataagc gctagtgtga       60

tcatgtccag aggagatatt gtgatgactc aaagccccga cagtctggcc gtgtctttgg      120

gcgagagagc cacaatcaac tgcaagtcct cacagagtat cctttattcc tctaatcaga      180

agaattacct cgcatggtat caacaaaaac ccggacagag ccctaagctt ttgatctatt      240

gggcatctac ccgagaatca ggagtgccgg accgcttcag tggatcagga tcaggcacag      300

actttacgct gacaatatcc tctcttcagg ccgaagacgt tgccgtgtac tactgccatc      360

aatatatgtc aagctacaca ttcggccagg gcaccaaact cgagattaag tctagtggtg      420

gcggaggcag tggcggagga ggctccgggg gcggagggtc ccaggttcag ctcgtgcaat      480

caggggcaga ggtcaagaag ccgggtgcct ctgtgaaggt gtcatgtaag gcctccgggt      540

atacttttgc cagcgcctgg atgcattgga tgaggcaggc gcccggccag ggtctggagt      600

ggattggttg gattaatccc ggaaacgtga atactaagta taacgagaag tttaagggca      660

gggccacact cacagtcgac acaagcacca ataccgcgta catggaactt tccagcctcc      720

ggtccgagga cactgcggtg tattactgcg cacgctccac ctattacaga ccacttgatt      780

actggggcca agggaccctg gtgaccgtgt ctagcgcact cagcaattcc atcatgtact      840

tctctcattt cgtgccagta tttctgcctg ccaagccaac taccacacct gcgccacgcc      900

ctcccacgcc cgcacccaca attgcttcac agcctctttc tctgcggcct gaggcttgtc      960

gcccagcagc cggaggcgcc gtgcatacgc gcggccttga cttcgcatgt gacatctaca     1020

tttgggctcc tttggctgga acctgcgggg tgttgttgct tagtctggtg attaccctct     1080

actgcaatca tagaaagcgg gggcgaaaga aacttctcta tatcttcaaa cagcctttca     1140

tgcgaccagt gcagacaacc caagaggaag acggatgcag ctgtcgcttt ccagaggagg     1200

aagaaggggg ctgcgagctg agagtgaaat tctctcgctc cgctgacgcc cccgcgtatc     1260

aacagggcca gaatcagctc tacaacgaac ttaaccttgg gcggagagaa gaatacgatg     1320

ttctcgacaa gcgcaggggg agagaccctg agatgggcgg gaaaccgcgc cgcaagaacc     1380

cccaagaagg gttgtataac gagctccaga aggacaaaat ggctgaagcc tactcagaga     1440

taggtatgaa gggcgagcgc cgcagaggga agggacacga tggtctgtac caaggccttt     1500

caaccgccac caaggatacc tatgatgcac tgcacatgca agccctgcct cctcgctaag     1560

gatcc                                                                 1565


<210>  19
<211>  67
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD28 transmembrane domain

<400>  19

Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn 
1               5                   10                  15      


Gly Thr Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu 
            20                  25                  30          


Phe Pro Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly 
        35                  40                  45              


Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe 
    50                  55                  60                  


Trp Val Arg 
65          


