HLA alleles in the context of RAMs in therapy naive patients Finja - - PowerPoint PPT Presentation

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Finja Schweitzer

Institute of Virology Cologne, Germany

HLA alleles in the context

• f RAMs in therapy naive patients

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Background

Human leukocyte antigen (HLA) class I molecules present viral peptides on infected cells to cytotoxic T cells (CTLs)

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

viral peptides are  8-10 amino acids long  defined for each HLA type (specific anker & recognition site)

Background

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

HLA types, which are associated with a high viremia and fast progression of disease  HLA-A1-B8-DR3, B37 due to:  ineffective presentation on HLA molecules  ineffective recognition by CTLs  immune escape mutants

Background

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

HLA class I molecule EG MN P GR W L

TCR

CTL

E E M NLP GR W

Antigen-presenting cell

T cell

HLA class I molecule

TCR

CTL Antigen-presenting cell

T cell

Background

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Antigen-presenting cell

T cell

E E M NLP GR W HLA class I molecule

TCR

CTL Antigen-presenting cell

T cell

E E M NLP Y R W HLA class I molecule

TCR

CTL

Background

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Escape Mutant Immune Response Inhibition Drug Resistance Infectious Agent: HIV Host: Human Therapy: Antiretrovirals

Background

Pressure of immune system and drugs on the same sequence:

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Aim

Influence of RAMs on the immune system  Identification of immune escape mutants among RAMs

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Methods and patients

• HLA-A, -B and –C typing of 139 RESINA patients

(with primary RAMs)

• Correlation of HLA alleles and mutations within PR

and RT

• Using epitope prediction server (netmhc) to identify

potential epitopes

• Analysis of clinical data

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results

PR: RT:

# of correlated mutations

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results 1 – Correlation PR

Specific mutations could be assigned to HLA alleles:

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results 1 – Correlation RT

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results 2: Epitope prediction PR

reference sequence: TGADDTVLEEMSLPGRWKPKM potential epitope: MSLPGRWK HLA-A*03: WB EMSLPGRWK HLA-A*03: WB MSLPGRWKPK HLA-A*03: WB MSLPGRWK HLA-A*30: WB MSLPGRWKPK HLA-A*30: SB reference sequence: GADDTVLEEMSLPGRWKPKMI potential epitope: EEMSLPGRW HLA-B*44: WB reference sequence: MSLPGRWKPKMIGGIGGFIKV potential epitope: MIGGIGGFIKV HLA-A*03: WB KMIGGIGGFIKV HLA-A*03: SB reference sequence: IGTVLVGPTPVNIIGRNLLTQ potential epitope: VLVGPTPV HLA-A*02: WB VLVGPTPVNI HLA-A*02: WB

Protease: M36I Protease: S37D/N Protease: M46I/L Protease: V82A

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results 2: Epitope prediction RT

reference sequence: EPFRKQNPDIVIYQYMDDLYV potential epitope: QNPDIVIY HLA-Cw*07: WB QNPDIVIYQY HLA-Cw*07: SB KQNPDIVIY HLA-Cw*07: WB KQNPDIVIYQY HLA-Cw*07: SB reference sequence: TKIEELRQHLLRWGLTTPDKK potential epitope: EELRQHLLRW HLA-B*44: WB reference sequence: YDPSKDLIAEIQKQGQGQWTY potential epitope: AEIQKQGQGQW HLA-B*44: WB

RT: V179D/E RT: L210F RT: I329L

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results 3: Clinical data analysis



CD4 cell count, viral load (at time point of 1st positive test)

 Δ

Days (time between 1st positive test and therapy start)

vs

Specific HLA type w/ specific mutation Specific HLA type w/out specific mutation

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results 3: Clinical data analysis

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results 4: Follow up

1st positive test 1st time point 2nd time point 3rd time point

vs

Specific HLA type w/ specific mutation Specific HLA type w/out specific mutation



CD4 cell count, VL (Median time between time points: 2-3 months)

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Results 4: Follow up

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Conclusion

• RAMs could be assigned to specific

HLA alleles

• Epitopes were predicted within regions
• f RAMs
• Trends could be seen between CD4 cell

counts / VL of two compared groups

University of Cologne Institute of Virology

10th AREVIR-GenaFor-Meeting Bonn, 2010, Finja Schweitzer

Thank you!

Jens Verheyen Maria Fraune Nadine Sichtig Eugen Schülter Claudia Müller Dörte Hammerschmidt Monika Timmen-Wego Saleta Sierra-Aragon Eva Heger Elena Knops Melanie Balduin Susanna Trapp Rolf Kaiser Herbert Pfister Martin Däumer Institute of Immunolgy and Genetics, Bernhard Thiele Kaiserslautern Thomas Harrer Department of Medicine III, Sandra Müller University Hospital of Erlangen Stefan Reuter

• Dept. of Gastroenterology,

Björn Jensen University of Düsseldorf Mark Oette Klösterchen Hospital, Cologne Stefan Esser Department of Dermatology, University Hospital of Essen HIV/HBV group Institute of Virology, University of Cologne

The RESINA-study group