adsorption and virus mu1 tiplication in liquid medium, while chondroitin sulfate was completely ineffective as an inhibitor. No diffference was noted in the behavior of the 2 variants with respect to their sensitivity toward the 4 sulfated polysaccharides.General application of the complement fixation technic for serologic diagnosis of infections caused by the varicella-zoster (V-Z) virus has been limited by difficulties in obtaining sufficiently potent antigens free from an ticomplemen tary activity.While fairly potent antigens can be prepared from vesicular fluids( 1) p the source is too undependable to meet the requirements of the diagnostic laboratory. In early studies on the V-Z virus, Weller and Witton( 2 ) showed that infected tissue cultures contained specific complement-fixing antigen, but concentration of the culture fluids by ultrafiltration was necessary to obtain antigens with sufficient reactivity. Concentrates prepared in this manner were anticomplementary, and while heating abolished this activity, it also reduced the antigenicity. Caunt et aZ(3) prepared complement-fixing antigens to the V-Z virus in cultures of human amnion cells; fluids from infected cultures were concentrated by drying from the frozen state, and the resulting antigens were free from anticomplementary activity.The procedures described to date for preparation of V-Z complement-fixing antigens *Supported by grant from Nat. Inst. of Allergy and Infect. Dis., U.S.P.H.S. from infected cell cultures are cumbersome, involving replacement of the culture medium a t relatively frequent intervals after infection as well as concentration of the antigenic material from large volumes of culture fluid. More importantly, despite such concentration the resulting antigens have been of low potency. This report describes a simple method for preparation of relatively high-titered antigens from the cellular phase of infected cultures of human fetal diploid skin and muscle cells. Materials and methods. Human fetal dip- Zoid cell strains. Initiation of cell strains:Cultures of human fetal diploid skin and muscle (HFDSM) cells were initiated from fetuses 2 to 5 months of age. Tissue from the limbs and torso of the fetus was minced and washed in 3 changes of Hanks' balanced salt solution (BSS). Cells were dispersed by overnight trypsinization at 4OC in 200 ml of a 0.25% trypsin solution, pH 7.5. The dispersed cells were centrifuged at 600 rpm for 20 minutes and the trypsin solution was removed. The cells were then resuspended in 4 0 ml of Hanks' BBS, transferred to a 50 ml graduated, conical contrifuge tube and centrifuged at 600 rpm for 20 minutes. The volume of the cell pack was noted and, after at UNSW Library on July 7, 2015 ebm.sagepub.com Downloaded from

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