Replication of herpes simplex virus in WI-38 cells was inhibited by phosphonoacetic acid, as measured by decreased virus cytopathogenic effect and incorporation of radiolabeled thymidine in virus-infected cells. The drug appeared to have no effect on adsorption, penetration, or release of the virus nor on the synthesis of ribonucleic acid or protein. It appeared to inhibit virus deoxyribonucleic acid synthesis.Herpes simplex virus (HSV) is a frequent infection in man with degrees of severity from mild to severe discomfort (cutaneous lesions) to severe infection damaging the eye sight (herpes keratitis) to severe and life-threatening diseases (herpes encephalitis). It has been suggested recently that herpes-viruses may be directly or indirectly associated with cancer in several animals: frog renal adenocarcinoma (4), malignant lymphomas in chickens (Marek's disease) (11), rabbits (6), and monkeys (8). Herpes-type viruses have been indirectly associated with Burkitt's lymphoma, nasopharyngeal carcinoma (5), and cervical carcinoma (1) in humans. 5-Iodo-2-deoxyuridine and cytosine arabinoside are two well-known nucleoside analogs and are inhibitors of herpervirus replication. Both compounds are quite toxic (2, 12), and 5-iodo-2-deoxyuridine has been shown to be carcinogenic and teratogenic (10).In earlier studies by Shipkowitz et al. (13), phosphonoacetic acid (PAA) was shown to be active against herpes dermatitis in mice and herpes keratitis in rabbits. We report here the efficacy of PAA against HSV in tissue culture, its effect on WI-38 cells, and studies related to the mechanism of action. Infected cells. Virus stock solutions were diluted in MM and were added to confluent WI-38 cells. The virus was allowed to adsorb for 1 h at 37 C. The medium and unadsorbed virus were removed by decanting and the cells were washed twice with MM. Cell cultures were then incubated at 37 C in MM.Drug tests. PAA was prepared from sodium phosphite and sodium chloroacetic acid as described by Nylen (9). Stock solutions were prepared in MM at a concentration of 10 mg/ml. Portions were then added aseptically to cell cultures to give the desired final concentrations of PAA.Ribonucleic acid (RNA), deoxyribonucleic acid (DNA), and protein synthesis. Cultures (1 ml) of WI-38 cells were grown and infected in tubes. At various intervals during the incubation, the cultures were pulse-labeled for 30 min with [14CIthymidine (0.5 MCi), [3Hjuridine (2.5 1Ci), or "4C-labeled amino acid mixtures (0.2 1Ci). PAA was added at the time of HSV adsorption and was continued in the culture medium thereafter. After each 30-min labeling with radioactive materials, triplicate tubes were removed, and then the medium was decanted. The cell layers were washed twice with cold MM. Then, 4 ml of phosphate-buffered saline were added to each tube, and the cells were scraped from the tube wall. Trichloroacetic acid was added to a final concentration of 5%. The mixture was frozen, thawed once, and

Liver enzyme levels, viral RNA, and the immune response against both structural and nonstructural hepatitis C virus (HCV) proteins have been studied in experimentally infected chimpanzees in order to further understand the natural history of HCV infection. An ELISA for measuring both IgG and IgM responses to core (c22), 33c (NS3), and c100 (NS4) was employed. The IgG response rates were 5/8 for core, and 8/8 for both 33c and c100. Utilizing this antigen combination, at least one antibody response is measureable at, or within 3 weeks of, the major ALT peak. Although no individual antibody response is universally associated with initial detection of seroconversion, the combination of all three recombinant proteins measures seroconversion an average of 54 days earlier than with c100 alone, in 6/8 of the animals. IgM responses were measureable in 5/8 of the chimpanzees, were of shorter duration, and usually arose concomitantly with IgG responses. IgM appears to be a good indicator of primary infection since neither boosting nor recrudescence of disease during the chronic phase of disease elicited a secondary IgM response. Viral RNA can be measured 4-7 days (average = 9 days) postinfection with the period preceding the ALT peak being characterized by several PCR positive segments interrupted by periods in which no viral RNA can be measured. Following the ALT peak, chronically infected animals with recurring ALT elevations are generally PCR positive with intercedent PCR negative periods. Those animals that appear to have biochemically resolved disease generally have PCR negative profiles, although they still may periodically exhibit PCR positive sera. This indicates that with the recent advent of new screening techniques, a more stringent definition of HCV resolution will be required.

Disodium phosphonoacetate when administered orally or topically to mice experimentally infected with herpes simplex virus was able to significantly reduce the mortality associated with the agent. In addition, this compound was able to reduce herpesvirus lesions on the corneas of infected rabbits.

Disodium phosphonoacetate when administered orally or topically to mice experimentally infected with herpes simplex virus was able to significantly reduce the mortality associated with the agent. In addition, this compound was able to reduce herpesvirus lesions on the corneas of infected rabbits.

Various multisurface mass-scale tissue culture propagators ranging in capacity up to 200 liters have been developed a t .4bbott Laboratories. These patented units consist of an enclosed vessel containing a multiplicity of separated glass plates or disks on which cells may attach and proliferate. Means for mixing and aeration of the medium are provided. Sample ports facilitate the addition of cultures and media, the withdrawal of samples, the washing of cell monolayers, and the harvesting of cells and cell products. The large cell growth surface per occupied volume, the provision for separating tissue cells from media simply and easily, and the minimization of the amount of labor required per cell growth area are some of the many advantages of the multisurface tissue propagator that are described.