1. Simian Foamy Viruses (SFV) Arifa S. Khan, Ph.D. DVP, OVRR, CBER, FDA BPAC, December 13, 2001

2. FV Classification Family: Retroviridae Genus: Spumavirus n Single-stranded RNA genome (12-13 kb) n tRNA lys1,2 primer for minus-strand synthesis n Integration is a critical event in the virus life cycle n Encode proteases and integrases Complex genome: gag, pol, env + several ORFsComplex genome: gag, pol, env + several ORFs Temporal regulation of transcriptionTemporal regulation of transcription Highly cytopathic with broad species-, tissue-, and cell-tropismHighly cytopathic with broad species-, tissue-, and cell-tropism Highly cell-associatedHighly cell-associated No evidence of pathogenicity in either natural or accidentally infected hostsNo evidence of pathogenicity in either natural or accidentally infected hosts

3. FV Classification Family: Hepadnaviridae Family: Hepadnaviridae Replication mechanism similar to complex DNA viruses: Replication mechanism similar to complex DNA viruses: multiple promoters (LTR, IP) multiple promoters (LTR, IP) Infectious particles have linear, DNA genomes Infectious particles have linear, DNA genomes Late reverse transcription Late reverse transcription Mature virions composed of two large Gag proteins Mature virions composed of two large Gag proteins Viral budding requires both Gag and Env Viral budding requires both Gag and Env Majority of virus buds through the ER. Most virus is Majority of virus buds through the ER. Most virus is intracellular. intracellular. Peristently infected cells contain large amounts of DNA Peristently infected cells contain large amounts of DNA (intracellular cycling pathway) (intracellular cycling pathway)

4. LTR prt-pol env tas orf-2 LTR gag SFV GENOME STRUCTURE

5. FV Replication Cycle rt ER ER budding budding CYTOPLASM NUCLEUS rt rt rt integ uncoat vDNA vRNA vCores ? ?

6. FV: Species Distribution Species Virus Designation Simian Simian foamy virus (SFV) Bovine Bovine syncytium-forming virus (BSFV; BFV) Feline Feline syncytium-forming virus (FeSFV; FFV) Murine Hamster syncytium-forming virus (HaSFV) Otariidine Sea lion foamy virus Ovine Sheep foamy virus Equine Horse foamy virus (EFV) Human Human foamy virus (HFV; SFVcpz[hu]; Acquired by cross-infection from a chimpanzee Acquired by cross-infection from a chimpanzee)

7. SFV Prevalence in Primates Species ProsimianProsimian Galago Galago New World PrimatesNew World Primates Squirrel, Spider, Capuchin, Marmoset Squirrel, Spider, Capuchin, Marmoset Old World PrimatesOld World Primates Macaques [Rhesus, Pigtailed, Cynomolgus, Macaques [Rhesus, Pigtailed, Cynomolgus, Formosan Rock macaque, Bonnet], Formosan Rock macaque, Bonnet], African green monkey; Mangabey; Baboon African green monkey; Mangabey; Baboon ApesApes Chimpanzee, Orangutan, Gorilla Chimpanzee, Orangutan, Gorilla

8. SFV Biology: Host Range and Cell Tropism Exceptionally broad host range Non-Primate Cell Type Tissue Origin Species Chicken, Quail Fibroblast Embyro Mouse Fibroblast Embryo, Tail Dog Fibroblast Thymus Cat Fibroblast Kidney

9. SFV Biology: Host Range and Cell Tropism Primate Cell Type Tissue Origin Species Monkey Fibroblast Lung, Kidney Epithelial Kidney Epithelial Kidney Macrophage Lung Macrophage Lung Human Fibroblast Lung Epithelial Lung, Muscle Epithelial Lung, Muscle Lymphoid T cells, B cells Lymphoid T cells, B cells Macrophage Macrophage Neural Neural

10. SFV Biology: In Vitro Replication SFV can replicate in all species Replication efficiency- (Mergia et al., J. Med. Primatol. 1996) l Cell type dependent (Mergia et al., J. Med. Primatol. 1996) Fibroblasts and Epithelial cells > Lymphoid and Fibroblasts and Epithelial cells > Lymphoid and Macrophage cells Macrophage cells (Khan et al., submitted) l Virus type dependent (Khan et al., submitted) Prototype SFV-1 and SFV-2 > Naturally-occurring Isolates (Khan et al., submitted) l Virus isolate dependent (Khan et al., submitted) SFV-1 > SFV-2 > Naturally-occurring Isolates

11. Latent Infection of A549 Human Tumor Cells with Naturally Occurring SFV Isolates

12. Lack of Virus Production in SFV Infected A549 Cells using STF-PERT Assay

13. PCR Detection of SFV Sequences in SFV Infected A549 Cells

14. SFV BIOLOGY: In Vitro Infection l Productive InfectionMost cell types especially fibroblasts l Productive Infection: Most cell types especially fibroblasts Variable amount of extracellular virus production - Variable amount of extracellular virus production (depending upon the species, tissue origin and (depending upon the species, tissue origin and cell type- Mus dunni cells most sensitive) cell type- Mus dunni cells most sensitive) - Cytopathic Effect- Lysis, Apoptosis - Cytopathic Effect- Lysis, Apoptosis l Chronic InfectionTransformed cell lines of myeloid, erythroid, and lymphoid origin l Chronic Infection: Transformed cell lines of myeloid, erythroid, and lymphoid origin - Low level virus production - No Cytopathic Effect - No Cytopathic Effect l Latent InfectionSelected cases l Latent Infection:Selected cases - No virus production - Virus induction/reactivation - Virus induction/reactivation

15. SFV Natural infection in Non-human Primates SFV is widespread in all non-human primate species Eleven serologically distinct subtypes (SFV-1 to SFV-11) Seroprevalence is high in capitivity (93% in AGM; Schweizer et al., 1995) Higher incidence in adults than infants (about 30% seropositive by 1 year of age in a baboon breeding colony- Blewett et al., 2000) Sequences genetically stable (minimal genetic drift over 13 years in AGM study- Schweizer et al., 1999) Broad tissue distribution (viral DNA persist in all tissues in AGM- Falcone et al., 1999) Latent, peristent infection (viral RNA detected only in oral mucosa- Falcone et al., 1999)

16. SFV Experimental Infection in Animals Immunocompetent rabbits and mice [SFVmac; SFVcpz; SFVcpz(hu)] Persistent infection Persistent infection Transient immunosuppressive effect Transient immunosuppressive effect No signs of any disease; no pathology No signs of any disease; no pathology In general, SFV infection in small animals was similar to naturally occurring infection in non- human primates.

17. SFV Experimental Infection in Animals Transgenic Mice [SFVcpz(hu)] Transgene expression in forebrain andTransgene expression in forebrain and cerbellum resulted in pathology in cerbellum resulted in pathology in tissues of the central nervous system tissues of the central nervous system and striated. Probably due to ORFs and striated. Probably due to ORFs (tas, bet). (tas, bet). Pathology enhanced in the presence ofPathology enhanced in the presence of structural genes. structural genes. FV replication not demonstrated.FV replication not demonstrated.

18. SFV “Accidental” Infection in Occupationally Exposed Humans SFV infection in non-human primate handlers and zoo keepers has occurred due to exposure to African green monkey, chimpanzee, baboon, macaque. Pesistent virus infection (>30 years in one animal handler) Latent virus infection (no evidence of plasma viremia; virus isolated in co-culture from peripheral blood lymphocytes). No evidence of virus transmission in close contacts. No clinical signs of FV-associated disease.

19. SFV Pathogenesis “A Virus in Search of a Disease” [Weiss, R.A., 1988, Nature (London) 333, 497-498] Tenuous association with various diseases in humans:Tenuous association with various diseases in humans: Thyroiditis de Quervain, Graves’ disease, Multiple sclerosis, Chronic fatigue syndrome, Familial Mediterranean fever, Senorineural hearing loss, Dialysis encephalopathy, and Myasthenia gravis. In most studies, use of multiple detection assay failed to confirm the initial disease associationIn most studies, use of multiple detection assay failed to confirm the initial disease association. In Myasthenia Gravis FV detected by serological and molecular assays but no infectious virus recovered. The authors suggested further studies be done to confirm the role of FV as an etiological agent in Myasthenia Gravis.

20. SFV Pathogenesis “A Virus in Search of a Disease” [Weiss, R.A. 1988 Nature (London) 333, 497-498] No evidence of any disease in non-human primates due to naturally occurring SFV. In small animal models using prototype, laboratory- adapted viruses, no disease seen in immunocompetent rabbits or mice; disease seen in transgenic mice due to protein expression. No evidence of disease in SFV-infected humans

21. SFV Summary SFV transmission is high in non human primatesSFV transmission is high in non human primates probably due to saliva. No disease. probably due to saliva. No disease. SFV can infect humans due to accidental exposureSFV can infect humans due to accidental exposure (mostly due to bites; other?). No evidence of human- (mostly due to bites; other?). No evidence of human- to-human transmission. No evidence of any virus- to-human transmission. No evidence of any virus- associated disease. associated disease. Experimental infection of mice and rabbitsExperimental infection of mice and rabbits demonstrated by different routes: intradermal, demonstrated by different routes: intradermal, intraperitoneal, intranasal. No disease in normal intraperitoneal, intranasal. No disease in normal animals. Disease in transgenic mice due to proteins. animals. Disease in transgenic mice due to proteins. n No evidence of FV transmission by blood due to lack of relevant animal studies. relevant animal studies.