The HSV 1 strain KOS and HSV 2 strain G were used as reference herpesviruses. Daudi cells were pifithrin a obtained from ATCC. All cell lines were cultured in RPMI 1640 medium supplemented with hands down the L glutamine and 10% fetal calf serum. The human embryonic lung fibroblasts, employed for the anti HSV assays, were obtained from the ATCC and cultured in minimal important medium supplemented with 0, 1000 Lglutamine and 10 percent FCS. Three full minutes sodium bicarbonate. The epithelial cell line HEC 1A was cultivated in DMEM medium supplemented with ten percent FCS and one of the HEPES. The VK2 epithelial cells were cultivated in serum free keratinocyte choice. Human astroglioma U87 cells expressing human CD4 were a kind present from Dr. Dan R. Littman. These cells were transfected with either CCR5 or CXCR4 and cultured in Dulbecco s modified Eagle s medium containing ten percent fetal bovine serum, 0. 01 M HEPES buffer, 0. 2 mg/ml geneticin and 1 mg/ml puromycin. Peripheral blood mononuclear cells were isolated from buffy coats from healthier blood donors, derived from the blood transfusion center, by density gradient centrifugation. The PBMCs were cultured in RPMI 1640 medium containing 10 % FCS and 1000 M glutamine. Stimulation was obtained pro-peptide by 2 mg/ml phytohemagglutinin for 3 days on 37uC before their further use within anti-hiv assays. Monocyte derived macrophages were prepared as follows: freshly isolated PBMCs were seeded in a 48 well plate in 1 ml RPMI 1640 medium with one hundred thousand FCS and incubated for 1 week at 37uC. A while later, the nonadherent cells were carefully mixed and removed from the adherent cell layer. The cells were watchfully cleaned and this washing step was repeated after 6 days of incubation at 37uC. Gemcitabine ic50 The adherent cells were then infected with HIV 1 R5 BaL. Infections The HIV 1 stresses NL4. HIV 1IIIB, 3 and HIV 1 BaL were received from the AIDS Research and Reference Reagent Program. The HIV 1 strain HE was later on cultured in a variety of CD4 T cell lines and initially isolated in the Rega Institute from the Belgian AIDS patient. Numerous HIV 1 clinical isolates, agent for different clades, were kindly supplied by Dr. J. L. Lathey and only passaged in PHA triggered PBMCs. Their coreceptor utilization was identified in house utilizing the U87. CD4. CXCR4 and U87. CD4. CCR5 transfected cells. The HIV 1 X4 isolate CI17 was obtained through the effort of the clinical AMD3100 trial via Dr. H. Bridger. HIV 2 ROD was received from the Medical Research Council. The in vitro generated HIV 1 IIIB or NL4. 3 strains resistant to raltegravir, AMD3100, enfuvirtide, 2G12 mAb, azidothymidinie and feglymycin were characterized in earlier in the day publications. Several HSV 1 wild type, HSV 1 thymidine kinase deficient, HSV 2 wild type and HSV 2 TK2 clinical isolates derived from virus infected people in Belgium were used.