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

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

• 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

Update from the AREVIR-projects

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Eva Heger/Rettenstein/2009

Conditions for MVC therapy?

VL 5 0

R5 / X4 ratio time

MVC R5 X4

50 VL

R5/ X4 ratio time

MVC

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Cell-Cell Fusion Assay

transfection

HIV-RNA gp120 PCR

LTR LTR LTR LTR gag gag pol pol env env

env expression pNL43

env expression vector

gp120 gp41 tat

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

CCR5+CXCR4

expressing cells

CXCR4

expressing cells

fusion

env expression

LTR luc LTR luc

LT R luc

+ +

Cell-Cell Fusion Assay

X4 R5

Luc activity

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

Cell-Cell Fusion Assay

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

X4 R5 X4 X4 X4 R5

Phenotype determination

SI NSI SI SI SI NSI

Cell culture Fusion assay

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Eva Heger/Rettenstein/2009

Summary

• Comparable results of fusion assay and

microscopic cell culture analysis

• Sensitivity for X4 detection ~ 10%

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Coreceptor analysis for MVC 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

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Trofile/geno2pheno

# of patients Trofile Geno2pheno

135

pos pos

22

neg neg

21

R5 16 D/M 5 neg

14

neg R5 9

6/RNA, 3/DNA

X4 5

5/RNA

133

VL<1000 neg 53 R5 52

25/RNA, 27/DNA

X4 28

12/RNA, 16/DNA

N = 325

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Number

• f pats.

Clinical response Trofile g2p comments

1

• Ø

X4

salvage 2

• R5

R5

no OBT 1

• D/ M

R5 RAL, T-20, DRV

19

+ R5 R5

11 *

+ Ø R5

7

+

VL< 1000

R5

therapy after g2p

2

+ R5 Ø

4

+ R5 X4

1

+ D/ M R5

1

+ D/ M X4

salvage

* 3 patients with proviral genotype only * 3 patients with proviral genotype only

Clinical response to MVC

N = 49

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Summary clinical data

• Coreceptor determination

– 28/49 patients Trofile™ and geno2pheno[coreceptor] – 2/49

patients Trofile™ only

– 19/49 patients geno2pheno[coreceptor] only

• Trofile™

– 25/27 patients

R5

successful outcome* – 2/3

patients D/M

successful outcome

• geno2pheno[coreceptor]

– 38/41 patients

R5

successful outcome * – 1/6

patient X4

successful outcome – 7/48

patients VL< 1000 R5 successful outcome

.* no backbone

University of Cologne Institute of Virology

AREVIR-Genafor-Meeting/ 2009/ Eva Heger

Thank you!

University of Cologne Gerd Fätkenheuer Institute of Virology Nadine Sichtig Saleta Sierra-Aragon Eva Heger Susanna Trapp Rolf Kaiser Herbert Pfister 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 Hospital of Bonn Bernd Kupfer Jürgen Rockstroh