A pleiotropic response is induced in F9 embryonal carcinoma cells and rhino mouse skin by All-trans-retinoic acid, a RAR agonist but not by SR11237, a RXR-selective agonist.

We evaluated SR11237, a retinoid X receptor (RXR)-specific compound, for its pharmacologic effects on cell differentiation in F9 embryonal carcinoma cells and rhino mouse epidermis. SR11237 can cause RXR/RXR homodimers to form and transactivate a reporter gene containing a RXR-response element. We confirmed, using nuclear receptor co-transfection assays in COS-1 cells, that SR11237 is effective at transactivating a chloramphenicol acetyltransferase reporter gene through RXRs but not retinoic acid receptors. When SR11237 was tested for its ability to modulate cell differentiation, it was inactive on F9 embryonal carcinoma cells and rhino mouse skin. Because differentiation in these systems is known to be regulated by RAR-specific compounds, such as all-trans-retinoic acid and (E)-4-[2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-1-prope nyl benzoic acid], our results with SR11237 are compatible with the concept that classical retinoid pleiotropic responses are mediated by RXR/RAR heterodimeric nuclear receptors rather than through RXR/RXR homodimers.

Dengue virus plaque development in simian cell systems. I. Factors influencing virus adsorption and variables in the agar overlay medium.

Chemical and physical variables influencing the plaquing of all dengue serotypes in two simian cell systems were studied. Calf serum in the nutrient overlay may be replaced by mouse ascitic fluid or bovine plasma albumin when employing the rhesus monkey kidney LLC-MK(2) cell system for plaquing all dengue serotypes. Doubling the serum concentration in the overlay had little effect in modifying dengue types 1, 2, 3, and 4 plaque titers. Newborn agamma, 4-week-old and 8-week-old calf serum gave comparable titers with all dengue virus serotypes. Dengue virus titers, plaque size, and development time were unaffected by sodium bicarbonate concentrations ranging from 1.1 to 4.4 mg/ml of overlay. A twofold increase (0.00332 g%) in the amount of either autoclaved or filtered-sterilized neutral red reduced the dengue 2 virus titer as much as 2.2 logs. An increased Mg(++) and decreased Ca(++) concentration in the overlay medium increased the efficiency of the plaquing system.

Dengue virus plaque development in simian cell systems. II. Agar variables and effect of chemical additives.

Dengue type 2 virus, strain New Guinea B, plaqued with equal facility and titer under overlays containing six different grades of commercial agar in the LLC-MK(2) cell system. Doubling the agar volume on LLC-MK(2) cell monolayers increased the plaque development time of dengue type 1, strain Hawaii. Storage of agar at 56 C reduced or totally abolished dengue type 4, strain H-241, plaque titer in LLC-MK(2) cells. The influence of six known virus plaque-enhancing compounds on plaque development of all dengue virus serotypes was studied in two continuous simian kidney cell lines, LLC-MK(2) and Vero. In the absence of any chemical additive, plaque development of all dengue serotypes was more rapid (4 to 10 days) in the LLC-MK(2) line than in the Vero line (6 to 13 days). Increased plaque development time of type 1, strain Hawaii, by pancreatin and plaque-size doubling of dengue types 1 and 4 was the only advantage conferred by the addition of six chemical additives in the LLC-MK(2) cell system. Dengue types 1 and 6 failed to plaque in the Vero cell system unless aided by a plaque-enhancing compound; plaques of dengue types 2, 3, 4, and 5 appeared sooner (2 days) and were increased in plaque diameter. The optimal DEAE concentration for plaquing dengue type 1, strain Hawaii, was 100 mug/ml; plaque development either failed at lower concentrations or was inhibited at higher (200 mug/ml) concentrations.

Arbovirus plaquing in a clonal line (PS Y-15) of porcine kidney.

Plaque development of 42 arbovirus serotypes, representing 17 serogroups, was studied in a clonal line (PS Y-15) of porcine kidney. The arboviruses plaqued within 3 to 7 days, with plaque diameters ranging from 1 to 5 mm. Mouse mortality and cell infectivity end points were compared with 24 arboviruses. Arbovirus LD(50) end points in mice exceeded the porcine kidney monolayer plaque-forming unitage end point with all 24 viruses compared.