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Cell cytotoxicity, inhibition of synthesis of Epstein-Barr virus (EBV) viral capsid antigen (VCA), nuclear antigen (EBNA) and transformation of human cord blood lymphocytes (CBL) by EBV, were studied using the disodium salt of phosphonoacetic acid (PAA), ethyl diethyl-phosphonoacetate (Et-PAA) and two derivatives of phosphonopropionic acid: ethyl diethyl-2-phosphonopropionate (Et-2-PPA) and ethyl diethyl-3-phosphonopropionate (Et-3-PPA). These substances were tested on EBV producing cell lines, B.95-8 and P3HR1. VCA and EBNA synthesis were determined by immunofluorescence and transformation of CBL by 3H-thymidine uptake. Up to 100 micrograms/ml of PAA were not toxic to 2.10(5) cells. Et-PAA, Et-2-PPA and Et-3-PPA were nontoxic at concentrations up to 2000 micrograms/ml. PAA inhibited 82.05% of EBV VCA synthesis at a concentration of 50 micrograms/ml. Et-PAA inhibited 47.01% of VCA at a concentration of 100 micrograms/ml and 78.09% at a concentration of 2000 micrograms/ml. Et-2-PPA inhibited 41.04% of VCA at 100 micrograms/ml and 80.87% when used at 2000 micrograms/ml. Et-3-PPA inhibited 35.06% of VCA at 100 micrograms/ml and 69.92% at 2000 micrograms/ml. Removal of the substances restored VCA synthesis. EBNA synthesis was not affected by these substances. PAA completely inhibited the transformation of human CBL by EBV at a concentration of 100 micrograms/ml. Et-PAA at a concentration of 2000 micrograms/ml completely inhibited 3H-thymidine uptake. ET-2-PPA was less effective whereas Et-3-PPA had almost no effects at the same concentration.