Correlations between transmitted HI V drug resistance mutations and - PowerPoint PPT Presentation

Correlations between transmitted HI V drug resistance mutations and HLA of therapy-naive HI V-patients 20.04.2007 | Martin Däumer, Institut für Virologie, Universität zu Köln / Labor Thiele, Kaiserslautern

Host Genetic Factors O’Brien SJ and Nelson GW. Human genes that limit AI DS. Nat Gen 2004 Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 2 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

I ntrahost Evolution ■ Evading host immune response ■ Developing drug resistance Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 3 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Escape from Humoral Immunity Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 4 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Escape from Cell Mediated Immunity Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 5 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Human Leukocyte Antigen (HLA) Human leukocyte antigen � displays peptides on cell surfaces in order to present antigens in the course of an adaptive immune response located on short arm of chromosome 6 � most polymorphic region of entire human genome HLA class I � associated with (viral) infections � interacts with cytotoxic T-cells (CTL) HLA class II � stimulates the helper T-cell activity HLA class III � encodes other components of the immune system e.g. complement components and cytokines Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 6 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Basic genetics to HLA Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 7 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Peptide processing and presentation Degradation of viral proteins inside cell into small peptides in cytosol � Peptides are loaded onto HLA molecule and transported to cell surface � Impaired presentation or processing of the peptides is due to mutations in CTL epitopes � cells are not destroyed by CTL Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 8 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Problem definition Do transmitted resistance mutations correlate with certain HLA types indicating escape from the immune system? Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 9 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Patients and Methods � 103 therapy-naive patients (RESINA) with documented drug resistance mutations in protease or / and reverse transcriptase � Determination of HLA-A and –B type by sequencing (n=75) � Frequencies of HLA-types were correlated with mutations in protease and reverse transcriptase � Significance of correlation determined using Fisher‘s exact probability test Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 10 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Ethnic analysis of the study cohort � The HLA-type is origin dependent . Or igin of P a t ie nt s origin unknown 8% Asiens 4% Africans 8% Caucasians 80% Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 11 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Mutations in RT Frequency of HLA-B*44 45,0% 41,7% 40,0% 35,0% Occurrence [%] 30,0% Frequencies of HLA-types in subgroups 25,0% compared to the whole group as well as 20,0% 15,0% literature values 9,3% 10,0% 5,8% 5,0% 0,0% Subgroup (K 103 Whole Group Literature Value R) n = 6; P = 0.0018 Frequency of HLA-B*07 Frequency of HLA-B*07 + HLA-B*40 Frequency of HLA-B*40 25,0% 20,0% 14,0% 17,6% 20,6% 11,8% 18,0% 12,0% 20,0% Occurrence [%] 16,0% Occurrence [%] Occurrence [%] 10,0% 14,0% 8,0% 15,0% 12,2% 12,0% 8,0% 6,7% 10,7% 10,0% 10,0% 6,0% 8,0% 5,3% 4,0% 6,0% 5,0% 4,0% 2,0% 0,8% 2,0% 0,0% 0,0% 0,0% Subgroup (V 118 Whole Group Literature Value Subgroup (V 118 Whole Group Literature Value Subgroup (V 118 Whole Group Literature Value I) I) I) n = 17; P = 0.4700 n = 17; P = 0.0344 n = 17; P = 0.0531 Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 12 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Mutations in RT Frequency of HLA-B*44 n = 5; P = 0.0074 45,0% 40,0% 40,0% 35,0% Occurrence [%] 30,0% 25,0% 20,0% 15,0% 9,3% 10,0% 5,8% 5,0% Frequency of HLA-A*11 0,0% Subgroup (L 210 Whole Group Literature Value 60,0% F) 50,0% 50,0% Occurrence [%] 40,0% 30,0% 20,0% 8,0% 6,2% 10,0% 0,0% n = 2; P = 0.0323 Subgroup (V 75 I) Whole Group Literature Value Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 13 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Mutations in PR Frequency of HLA-B*35 Frequency of HLA-A*01 30,0% 37,5% 25,0% 40,0% Occurrence [%] 25,0% 35,0% Occurrence [%] 30,0% 20,0% 14,7% 25,0% 15,0% 20,0% 15,2% 9,7% 8,5% 12,7% 10,0% 15,0% 10,0% 5,7% 5,0% 5,0% 0,0% 0,0% Subgroup (L Whole Group Literature Literature Subgroup (L Whole Group Literature Literature 33 F) Value for Value for 33 F) Value for Value for Caucasian African Caucasian African n = 4; P = 0.6044 n = 4; P = 0.0643 Population Population Population Population Frequency of HLA-A*01 + HLA-B*35 60,0% 50,0% Occurrence [%] 50,0% 40,0% 30,0% 20,0% 5,3% 10,0% 1,5% 0,5% 0,0% Subgroup (L Whole Group Literature Literature 33 F) Value for Value for Caucasian African n = 4; P = 0.0125 Population Population Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 14 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Mutations in PR Frequency of HLA-A*03 Frequency of HLA-B*35 45,0% 45,0% 40,0% 40,0% 40,0% 40,0% 35,0% Occurrence [%] 35,0% Occurrence [%] 30,0% 30,0% 25,0% 25,0% 20,0% 20,0% 14,7% 14,0% 13,4% 15,0% 15,0% 9,7% 10,0% 10,0% 5,0% 5,0% 0,0% 0,0% Subgroup (M 46 I Whole Group Literature Value Subgroup (M 46 I Whole Group Literature Value / L) / L) n = 5; P = 0.0344 n = 5; P = 0.0406 Frequency of HLA-A*03 + HLA-B*35 70,0% 60,0% 60,0% Occurrence [%] 50,0% 40,0% 30,0% 20,0% 12,0% 10,0% 1,3% 0,0% Subgroup (M 46 I Whole Group Literature Value / L) n = 5; P = 0.0109 Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 15 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Correlation of resistance mutations and HLA types Frequency in genotype Mutation HLA-A / -B Lit. Value P value Subgroup Whole group K103R B* 44 41.7 % 9.3 % 5.8 % 0.0018 V118I B* 07 + B* 40 17.6 % 5.3 % 0.8 % 0.0344 L210F B* 44 40 % 9.3 % 5.8 % 0.0074 V75I A* 11 50 % 8 % 6.2 % 0.0323 1.5 % 0.5 % L33F A* 01 + B* 35 50% 5.3 % 0.0125 Caucasians Africans M46I/L A* 03 40 % 14 % 13.4 % 0.0344 M46I/L B* 35 40 % 14.7 % 9.7 % 0.0406 M46I/L A* 03 + B* 35 60% 12 % 1.3 % 0.0109 Phenotypic HLA frequency: presence of the HLA molecule on the cell surface, independent of the gene dose Genotypic HLA frequency: presence of the HLA molecule multiplied by the gene dose Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 16 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

I s this cohort representative? Control Investigation of the already reported correlation D121x & HLA-B* 35 (P = 0.0001) (Moore et al ., 2002) � Our study revealed a P value of: 0.000005 � proves that this study cohort acts like a normal population of HIV positive patients and the previous selection of patients did not cause a shift (bias) in the normal distribution of variation Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 17 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern

Summary I Ethnic analysis � 80.5 % of Caucasian origin � 7.8 % of African origin � 3.9 % of Asian origin � 7.8 % origin unknown 75/ 103 ART naive patients who harbor HIV drug resistance mutations were typed for both HLA-A & -B for 8 resistance associated positions not yet reported significant correlations between drug resistance mutations and HLA-type were found: K103R & HLA-B* 44; V118I & HLA-B* 07 + HLA-B* 40; L210F & HLA-B* 44; V75I & HLA-A* 11; L33F & HLA-A* 01 + HLA-B* 35; M46I/L & HLA-A* 03; M46I/L & HLA-B* 35; M46I/L & HLA-A* 03 + HLA-B* 35 Institut fü Institut f ür Immunologie und Genetik r Immunologie und Genetik Seite 18 20.04.2007 | Martin Däumer Medizinisches Labor Kaiserslautern Medizinisches Labor Kaiserslautern