               SEQUENCE LISTING

<110> Medizinische Hochschule Hannover
      Technische Universität Braunschweig

<120> CAR for use in the treatment of HvG disease

<130> M1085PCT

<150> EP19166421.8
<151> 2019-03-29

<160> 11

<170> BiSSAP 1.3.6

<210> 1
<211> 81
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD8 hinge domain and transmembran domain

<400> 1
Phe Val Pro Val Phe Leu Pro Ala Lys Pro Thr Thr Thr Pro Ala Pro 
1               5                   10                  15      
Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu 
            20                  25                  30          
Arg Pro Glu Ala Ser Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg 
        35                  40                  45              
Gly Leu Asp Phe Ala Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr 
    50                  55                  60                  
Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Asn His 
65                  70                  75                  80  
Arg 
    

<210> 2
<211> 42
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD28 signalling domain

<400> 2
Trp Val Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn 
1               5                   10                  15      
Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr 
            20                  25                  30          
Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg 
        35                  40          

<210> 3
<211> 113
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD3zeta signalling domain

<400> 3
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         
Ser 
    

<210> 4
<211> 280
<212> PRT
<213> Artificial Sequence


<220> 
<223> hΔFC IgG domain

<400> 4
Met Gly Ala Gly Ala Thr Gly Arg Ala Met Asp Gly Pro Arg Leu Leu 
1               5                   10                  15      
Leu Leu Leu Leu Leu Gly Val Ser Leu Gly Gly Ala Lys Glu Ala Cys 
            20                  25                  30          
Pro Thr Gly Leu Tyr Thr His Ser Gly Glu Cys Cys Lys Ala Cys Asn 
        35                  40                  45              
Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala Asn Gln Thr Val Cys 
    50                  55                  60                  
Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp Val Val Ser Ala Thr 
65                  70                  75                  80  
Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly Leu Gln Ser Met Ser 
                85                  90                  95      
Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys Arg Cys Ala Tyr Gly 
            100                 105                 110         
Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu Ala Cys Arg Val Cys 
        115                 120                 125             
Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln Asp Lys Gln Asn Thr 
    130                 135                 140                 
Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser Asp Glu Ala Asn His 
145                 150                 155                 160 
Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu Asp Thr Glu Arg Gln 
                165                 170                 175     
Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu Cys Glu Glu Ile Pro 
            180                 185                 190         
Gly Arg Trp Ile Thr Arg Ser Thr Pro Pro Glu Gly Ser Asp Ser Thr 
        195                 200                 205             
Ala Pro Ser Thr Gln Glu Pro Glu Ala Pro Pro Glu Gln Asp Leu Ile 
    210                 215                 220                 
Ala Ser Thr Val Ala Gly Val Val Thr Thr Val Met Gly Ser Ser Gln 
225                 230                 235                 240 
Pro Val Val Thr Arg Gly Thr Thr Asp Asn Leu Ile Pro Val Tyr Cys 
                245                 250                 255     
Ser Ile Leu Ala Ala Val Val Val Gly Leu Val Ala Tyr Ile Ala Phe 
            260                 265                 270         
Lys Arg Trp Asn Arg Gly Ile Leu 
        275                 280 

<210> 5
<211> 184
<212> PRT
<213> Artificial Sequence


<220> 
<223> hCD28 transmembrane domain - hCD3zeta signalling domain

<400> 5
Asp Pro Lys Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys 
1               5                   10                  15      
Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser 
            20                  25                  30          
Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg 
        35                  40                  45              
Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Ala Tyr Ala Ala Ala Arg 
    50                  55                  60                  
Asp Phe Ala Ala Tyr Arg Ser Leu Arg Val Lys Phe Ser Arg Ser Ala 
65                  70                  75                  80  
Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu 
                85                  90                  95      
Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly 
            100                 105                 110         
Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu 
        115                 120                 125             
Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser 
    130                 135                 140                 
Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly 
145                 150                 155                 160 
Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu 
                165                 170                 175     
His Met Gln Ala Leu Pro Pro Arg 
            180                 

<210> 6
<211> 452
<212> PRT
<213> Artificial Sequence


<220> 
<223> fusion P2A hFOXP3

<400> 6
Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu 
1               5                   10                  15      
Asn Pro Gly Pro Ser Met Pro Asn Pro Arg Pro Gly Lys Pro Ser Ala 
            20                  25                  30          
Pro Ser Leu Ala Leu Gly Pro Ser Pro Gly Ala Ser Pro Ser Trp Arg 
        35                  40                  45              
Ala Ala Pro Lys Ala Ser Asp Leu Leu Gly Ala Arg Gly Pro Gly Gly 
    50                  55                  60                  
Thr Phe Gln Gly Arg Asp Leu Arg Gly Gly Ala His Ala Ser Ser Ser 
65                  70                  75                  80  
Ser Leu Asn Pro Met Pro Pro Ser Gln Leu Gln Leu Pro Thr Leu Pro 
                85                  90                  95      
Leu Val Met Val Ala Pro Ser Gly Ala Arg Leu Gly Pro Leu Pro His 
            100                 105                 110         
Leu Gln Ala Leu Leu Gln Asp Arg Pro His Phe Met His Gln Leu Ser 
        115                 120                 125             
Thr Val Asp Ala His Ala Arg Thr Pro Val Leu Gln Val His Pro Leu 
    130                 135                 140                 
Glu Ser Pro Ala Met Ile Ser Leu Thr Pro Pro Thr Thr Ala Thr Gly 
145                 150                 155                 160 
Val Phe Ser Leu Lys Ala Arg Pro Gly Leu Pro Pro Gly Ile Asn Val 
                165                 170                 175     
Ala Ser Leu Glu Trp Val Ser Arg Glu Pro Ala Leu Leu Cys Thr Phe 
            180                 185                 190         
Pro Asn Pro Ser Ala Pro Arg Lys Asp Ser Thr Leu Ser Ala Val Pro 
        195                 200                 205             
Gln Ser Ser Tyr Pro Leu Leu Ala Asn Gly Val Cys Lys Trp Pro Gly 
    210                 215                 220                 
Cys Glu Lys Val Phe Glu Glu Pro Glu Asp Phe Leu Lys His Cys Gln 
225                 230                 235                 240 
Ala Asp His Leu Leu Asp Glu Lys Gly Arg Ala Gln Cys Leu Leu Gln 
                245                 250                 255     
Arg Glu Met Val Gln Ser Leu Glu Gln Gln Leu Val Leu Glu Lys Glu 
            260                 265                 270         
Lys Leu Ser Ala Met Gln Ala His Leu Ala Gly Lys Met Ala Leu Thr 
        275                 280                 285             
Lys Ala Ser Ser Val Ala Ser Ser Asp Lys Gly Ser Cys Cys Ile Val 
    290                 295                 300                 
Ala Ala Gly Ser Gln Gly Pro Val Val Pro Ala Trp Ser Gly Pro Arg 
305                 310                 315                 320 
Glu Ala Pro Asp Ser Leu Phe Ala Val Arg Arg His Leu Trp Gly Ser 
                325                 330                 335     
His Gly Asn Ser Thr Phe Pro Glu Phe Leu His Asn Met Asp Tyr Phe 
            340                 345                 350         
Lys Phe His Asn Met Arg Pro Pro Phe Thr Tyr Ala Thr Leu Ile Arg 
        355                 360                 365             
Trp Ala Ile Leu Glu Ala Pro Glu Lys Gln Arg Thr Leu Asn Glu Ile 
    370                 375                 380                 
Tyr His Trp Phe Thr Arg Met Phe Ala Phe Phe Arg Asn His Pro Ala 
385                 390                 395                 400 
Thr Trp Lys Asn Ala Ile Arg His Asn Leu Ser Leu His Lys Cys Phe 
                405                 410                 415     
Val Arg Val Glu Ser Glu Lys Gly Ala Val Trp Thr Val Asp Glu Leu 
            420                 425                 430         
Glu Phe Arg Lys Lys Arg Ser Gln Arg Pro Ser Arg Cys Ser Asn Pro 
        435                 440                 445             
Thr Pro Gly Pro 
    450         

<210> 7
<211> 280
<212> PRT
<213> Artificial Sequence


<220> 
<223> ΔLNGFR (reporter)

<400> 7
Met Gly Ala Gly Ala Thr Gly Arg Ala Met Asp Gly Pro Arg Leu Leu 
1               5                   10                  15      
Leu Leu Leu Leu Leu Gly Val Ser Leu Gly Gly Ala Lys Glu Ala Cys 
            20                  25                  30          
Pro Thr Gly Leu Tyr Thr His Ser Gly Glu Cys Cys Lys Ala Cys Asn 
        35                  40                  45              
Leu Gly Glu Gly Val Ala Gln Pro Cys Gly Ala Asn Gln Thr Val Cys 
    50                  55                  60                  
Glu Pro Cys Leu Asp Ser Val Thr Phe Ser Asp Val Val Ser Ala Thr 
65                  70                  75                  80  
Glu Pro Cys Lys Pro Cys Thr Glu Cys Val Gly Leu Gln Ser Met Ser 
                85                  90                  95      
Ala Pro Cys Val Glu Ala Asp Asp Ala Val Cys Arg Cys Ala Tyr Gly 
            100                 105                 110         
Tyr Tyr Gln Asp Glu Thr Thr Gly Arg Cys Glu Ala Cys Arg Val Cys 
        115                 120                 125             
Glu Ala Gly Ser Gly Leu Val Phe Ser Cys Gln Asp Lys Gln Asn Thr 
    130                 135                 140                 
Val Cys Glu Glu Cys Pro Asp Gly Thr Tyr Ser Asp Glu Ala Asn His 
145                 150                 155                 160 
Val Asp Pro Cys Leu Pro Cys Thr Val Cys Glu Asp Thr Glu Arg Gln 
                165                 170                 175     
Leu Arg Glu Cys Thr Arg Trp Ala Asp Ala Glu Cys Glu Glu Ile Pro 
            180                 185                 190         
Gly Arg Trp Ile Thr Arg Ser Thr Pro Pro Glu Gly Ser Asp Ser Thr 
        195                 200                 205             
Ala Pro Ser Thr Gln Glu Pro Glu Ala Pro Pro Glu Gln Asp Leu Ile 
    210                 215                 220                 
Ala Ser Thr Val Ala Gly Val Val Thr Thr Val Met Gly Ser Ser Gln 
225                 230                 235                 240 
Pro Val Val Thr Arg Gly Thr Thr Asp Asn Leu Ile Pro Val Tyr Cys 
                245                 250                 255     
Ser Ile Leu Ala Ala Val Val Val Gly Leu Val Ala Tyr Ile Ala Phe 
            260                 265                 270         
Lys Arg Trp Asn Arg Gly Ile Leu 
        275                 280 

<210> 8
<211> 21
<212> PRT
<213> Artificial Sequence


<220> 
<223> secretory leader peptide

<400> 8
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 
1               5                   10                  15      
His Ala Ala Ala Pro 
            20      

<210> 9
<211> 246
<212> PRT
<213> Artificial Sequence


<220> 
<221> CHAIN
<222> 1..123
<223> heavy chain variable region

<220> 
<223> scFv (ELFA*2)

<220> 
<221> CHAIN
<222> 26..33
<223> CDR1

<220> 
<221> CHAIN
<222> 51..58
<223> CDR2

<220> 
<221> CHAIN
<222> 97..112
<223> CDR3

<220> 
<221> CHAIN
<222> 124..139
<223> linker

<220> 
<221> CHAIN
<222> 140..246
<223> light chain variable kappa

<220> 
<221> CHAIN
<222> 166..171
<223> CDR1

<220> 
<221> CHAIN
<222> 189..191
<223> CDR2

<220> 
<221> CHAIN
<222> 228..237
<223> CDR3

<400> 9
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Gly 
1               5                   10                  15      
Ser Leu Arg Val Ser Cys Ala Ala Ser Gly Val Thr Leu Ser Asp Tyr 
            20                  25                  30          
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 
        35                  40                  45              
Ala Phe Ile Arg Asn Asp Gly Ser Asp Lys Tyr Tyr Ala Asp Ser Val 
    50                  55                  60                  
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Glu Lys Thr Val Ser 
65                  70                  75                  80  
Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 
                85                  90                  95      
Ala Lys Asn Gly Glu Ser Gly Pro Leu Asp Tyr Trp Tyr Leu Asp Leu 
            100                 105                 110         
Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser 
        115                 120                 125             
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Asp Val Val Met Thr 
    130                 135                 140                 
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile 
145                 150                 155                 160 
Thr Cys Gln Ser Ser Leu Asp Ile Ser His Tyr Leu Asn Trp Tyr Gln 
                165                 170                 175     
Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Asn 
            180                 185                 190         
Leu Glu Thr Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr 
        195                 200                 205             
His Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr 
    210                 215                 220                 
Tyr Tyr Cys Gln Gln Tyr Asp Asn Leu Pro Leu Thr Phe Gly Gly Gly 
225                 230                 235                 240 
Thr Lys Leu Glu Ile Lys 
                245     

<210> 10
<211> 21
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD28 extracellular domain signal peptide

<400> 10
Met Ala Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 
1               5                   10                  15      
His Ala Ala Ala Pro 
            20      

<210> 11
<211> 245
<212> PRT
<213> Artificial Sequence


<220> 
<221> CHAIN
<222> 1..123
<223> heavy chain variable region

<220> 
<223> variant ELFA*2

<220> 
<221> CHAIN
<222> 124..138
<223> linker

<220> 
<221> CHAIN
<222> 139..245
<223> light chain varible kappa

<400> 11
Gln Val Gln Leu Val Gln Ser Gly Gly Gly Val Val Gln Pro Gly Gly 
1               5                   10                  15      
Ser Leu Arg Val Ser Cys Ala Ala Ser Gly Val Thr Leu Ser Asp Tyr 
            20                  25                  30          
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 
        35                  40                  45              
Ala Phe Ile Arg Asn Asp Gly Ser Asp Lys Tyr Tyr Ala Asp Ser Val 
    50                  55                  60                  
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Glu Lys Thr Val Ser 
65                  70                  75                  80  
Leu Gln Met Ser Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 
                85                  90                  95      
Ala Lys Asn Gly Glu Ser Gly Pro Leu Asp Tyr Trp Tyr Leu Asp Leu 
            100                 105                 110         
Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser 
        115                 120                 125             
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Val Val Met Thr Gln 
    130                 135                 140                 
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 
145                 150                 155                 160 
Cys Gln Ser Ser Leu Asp Ile Ser His Tyr Leu Asn Trp Tyr Gln Gln 
                165                 170                 175     
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Asn Leu 
            180                 185                 190         
Glu Thr Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr His 
        195                 200                 205             
Phe Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 
    210                 215                 220                 
Tyr Cys Gln Gln Tyr Asp Asn Leu Pro Leu Thr Phe Gly Gly Gly Thr 
225                 230                 235                 240 
Lys Leu Glu Ile Lys 
                245 

