Coreceptor Tropism of HIV-1 Development of a virus-free Assay HIV-1 - PowerPoint PPT Presentation

Update from the AREVIR-projects University of Cologne Institute of Virology •Coreceptor Tropism of HIV-1 Development of a virus-free Assay •HIV-1 Tropism Study Comparison of Trofile™ and Geno2pheno [coreceptor] Eva Heger Institute of Virology, University of Cologne AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Conditions for MVC therapy? University of Cologne Institute of Virology MVC R5 / X4 VL R5 ratio X4 5 0 time MVC R5/ X4 VL ratio 50 time Eva Heger/Rettenstein/2009 AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Cell-Cell Fusion Assay University of Cologne Institute of Virology pNL43 transfection gp120 gp41 tat env expression vector env expression gp120 PCR HIV-RNA LTR LTR gag gag pol pol env env LTR LTR AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Cell-Cell Fusion Assay University of Cologne Institute of Virology CCR5+CXCR4 X4 R5 expressing cells LT luc R LTR luc + fusion Luc activity + env expression LTR luc CXCR4 expressing cells AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Cell-Cell Fusion Assay University of Cologne Institute of Virology AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Cell-Cell Fusion Assay University of Cologne Institute of Virology AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Phenotype determination University of Cologne Institute of Virology Fusion assay X4 R5 X4 X4 X4 R5 Cell culture SI NSI SI SI SI NSI AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Summary University of Cologne Institute of Virology • Comparable results of fusion assay and microscopic cell culture analysis • Sensitivity for X4 detection ~ 10% Eva Heger/Rettenstein/2009 AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Coreceptor analysis for MVC University of Cologne Institute of Virology administration • VL< 1000 copies/ml as limiting factor for phenotypical coreceptor analysis by Trofile ™ • MVC treatment history – till June 2008 patients mostly with VL > 1000 (only 22.9% patients with VL< 1000) – nowadays, 51.2% patients with VL< 1000 AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Trofile/geno2pheno University of Cologne Institute of Virology # of patients Trofile Geno2pheno 135 pos pos 22 neg neg R5 � 16 21 neg D/M � 5 R5 � 9 14 neg 6/RNA, 3/DNA X4 � 5 5/RNA neg � 53 133 VL<1000 R5 � 52 25/RNA, 27/DNA X4 � 28 12/RNA, 16/DNA N = 325 AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Clinical response to MVC University of Cologne Institute of Virology Number Clinical Trofile g2p comments of pats. response - Ø X4 1 salvage - R5 R5 2 no OBT - D/ M RAL, T-20, DRV 1 R5 + R5 R5 19 + Ø R5 11 * + R5 7 VL< 1000 therapy after g2p + R5 Ø 2 + R5 X4 4 + D/ M R5 1 + D/ M X4 1 salvage * 3 patients with proviral genotype only * 3 patients with proviral genotype only N = 49 AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Summary clinical data University of Cologne Institute of Virology • Coreceptor determination – 28/49 patients Trofile™ and geno2pheno [coreceptor] – 2/49 patients Trofile™ only – 19/49 patients geno2pheno [coreceptor] only • Trofile™ – 25/27 patients � � successful outcome * R5 patients � � successful outcome – 2/3 D/M • geno2pheno [coreceptor] – 38/41 patients � � successful outcome * R5 patient � � successful outcome – 1/6 X4 patients � VL< 1000 � R5 � successful outcome – 7/48 AREVIR-Genafor-Meeting/ 2009/ Eva Heger .* no backbone

Thank you! University of Cologne Institute of Virology NRZ University of Erlangen Sabine Wittmann Hauke Walter MPI for Informatics, Saarbrücken Alexander Thielen Thomas Lengauer University of Duisburg-Essen Stefan Esser University Hospital of Düsseldorf Stefan Reuter Björn Jensen University of Cologne Gerd Fätkenheuer Institute of Virology University Hospital of Bonn Nadine Sichtig Bernd Kupfer Saleta Sierra-Aragon Jürgen Rockstroh Eva Heger Susanna Trapp Rolf Kaiser Herbert Pfister AREVIR-Genafor-Meeting/ 2009/ Eva Heger