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TRIM5 suppresses cross-species transmission of a primate immunodeficiency virus and selects for emergence of resistant variants in the new species.

Kirmaier A, Wu F, Newman RM, Hall LR, Morgan JS, O'Connor S, Marx PA, Meythaler M, Goldstein S, Buckler-White A, Kaur A, Hirsch VM, Johnson WE - PLoS Biol. (2010)

Bottom Line: Simian immunodeficiency viruses of sooty mangabeys (SIVsm) are the source of multiple, successful cross-species transmissions, having given rise to HIV-2 in humans, SIVmac in rhesus macaques, and SIVstm in stump-tailed macaques.Surprisingly, transmission occurred even in individuals bearing restrictive TRIM5 genotypes, resulting in attenuation of replication rather than an outright block to infection.In cell-culture assays, the same TRIM5 alleles associated with viral suppression in vivo blocked infectivity of two SIVsm strains, but not the macaque-adapted strain SIVmac239.

View Article: PubMed Central - PubMed

Affiliation: New England Primate Research Center, Department of Microbiology and Molecular Genetics, Harvard Medical School, Southborough, Massachusetts, United States of America.

ABSTRACT
Simian immunodeficiency viruses of sooty mangabeys (SIVsm) are the source of multiple, successful cross-species transmissions, having given rise to HIV-2 in humans, SIVmac in rhesus macaques, and SIVstm in stump-tailed macaques. Cellular assays and phylogenetic comparisons indirectly support a role for TRIM5alpha, the product of the TRIM5 gene, in suppressing interspecies transmission and emergence of retroviruses in nature. Here, we investigate the in vivo role of TRIM5 directly, focusing on transmission of primate immunodeficiency viruses between outbred primate hosts. Specifically, we retrospectively analyzed experimental cross-species transmission of SIVsm in two cohorts of rhesus macaques and found a significant effect of TRIM5 genotype on viral replication levels. The effect was especially pronounced in a cohort of animals infected with SIVsmE543-3, where TRIM5 genotype correlated with approximately 100-fold to 1,000-fold differences in viral replication levels. Surprisingly, transmission occurred even in individuals bearing restrictive TRIM5 genotypes, resulting in attenuation of replication rather than an outright block to infection. In cell-culture assays, the same TRIM5 alleles associated with viral suppression in vivo blocked infectivity of two SIVsm strains, but not the macaque-adapted strain SIVmac239. Adaptations appeared in the viral capsid in animals with restrictive TRIM5 genotypes, and similar adaptations coincide with emergence of SIVmac in captive macaques in the 1970s. Thus, host TRIM5 can suppress viral replication in vivo, exerting selective pressure during the initial stages of cross-species transmission.

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Differential restriction of SIVmac and SIVsm strains by multiple alleles of rhesus macaque TRIM5.Single-cycle infectivity was measured on a panel of cell lines stably expressing six common alleles of rhesus TRIM5 [15],[17]. Alleles tested included three TRIM5TFP (Mamu-1, Mamu-2, Mamu-3, dark blue bars), two TRIM5Q (Mamu-4 and Mamu-5, light blue bars), and TRIM5CypA (orange bars). Control cells stably expressing the empty vector served as a negative control (black bars). Of the four closely related SIVs, only the rhesus macaque isolate SIVmac239 is resistant to multiple alleles. Infectivity (% GFP-positive cells) was measured by flow-cytometry (error bars indicate ± SEM). Virus stocks were first titered by serial dilution on parental cells and normalized (Figure S1). Stable cell lines were generated from CRFK cells as described in the Materials and Methods section. HIV-1NL4-3 was used as a positive control to confirm expression and function of the TRIM5Q/Q alleles. (A) SIVmac239; (B) SIVsmE041; (C) SIVsmE543-3; (D) SIVstm37/13; (E) HIV-1NL4-3; (F) Immunoblot confirming expression of HA-tagged TRIM5 proteins in cell lysates. upper panel, TRIM5 proteins; lower panel, β-actin loading control. Lanes: (1) Mamu-1; (2) Mamu-2; (3) Mamu-3; (4) Mamu-4; (5) Mamu-5; M, protein standard; TC, TRIM5CypA; V, control cells expressing vector-only. GenBank accession number for the SIVsmE041 clone: HM059825.
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pbio-1000462-g002: Differential restriction of SIVmac and SIVsm strains by multiple alleles of rhesus macaque TRIM5.Single-cycle infectivity was measured on a panel of cell lines stably expressing six common alleles of rhesus TRIM5 [15],[17]. Alleles tested included three TRIM5TFP (Mamu-1, Mamu-2, Mamu-3, dark blue bars), two TRIM5Q (Mamu-4 and Mamu-5, light blue bars), and TRIM5CypA (orange bars). Control cells stably expressing the empty vector served as a negative control (black bars). Of the four closely related SIVs, only the rhesus macaque isolate SIVmac239 is resistant to multiple alleles. Infectivity (% GFP-positive cells) was measured by flow-cytometry (error bars indicate ± SEM). Virus stocks were first titered by serial dilution on parental cells and normalized (Figure S1). Stable cell lines were generated from CRFK cells as described in the Materials and Methods section. HIV-1NL4-3 was used as a positive control to confirm expression and function of the TRIM5Q/Q alleles. (A) SIVmac239; (B) SIVsmE041; (C) SIVsmE543-3; (D) SIVstm37/13; (E) HIV-1NL4-3; (F) Immunoblot confirming expression of HA-tagged TRIM5 proteins in cell lysates. upper panel, TRIM5 proteins; lower panel, β-actin loading control. Lanes: (1) Mamu-1; (2) Mamu-2; (3) Mamu-3; (4) Mamu-4; (5) Mamu-5; M, protein standard; TC, TRIM5CypA; V, control cells expressing vector-only. GenBank accession number for the SIVsmE041 clone: HM059825.

Mentions: Infectivity of all four viruses was measured on cell lines stably expressing six common alleles of rhesus macaque TRIM5, including three TRIM5TFP alleles, two TRIM5Q alleles, and the TRIM5CypA allele (Figure 2). SIVmac239 was resistant to all six. SIVsmE041, SIVsmE543, and SIVstm were also resistant to TRIM5Q but unlike SIVmac239 were sensitive to both TRIM5CypA alleles and TRIM5TFP alleles (Figure 2B,C,D). Of the four SIV strains tested, only SIVmac239 is the product of decades of replication and spread in rhesus macaques. Thus, the comparison suggests that sensitivity to TRIM5TFP and TRIM5CypA alleles represents the ancestral phenotype and that emergence of SIVsm (as SIVmac) in rhesus macaques required acquisition of adaptive changes to overcome those particular types of alleles. In contrast, the SIVsm variants that first invaded rhesus macaques were probably inherently resistant to TRIM5Q alleles.


TRIM5 suppresses cross-species transmission of a primate immunodeficiency virus and selects for emergence of resistant variants in the new species.

Kirmaier A, Wu F, Newman RM, Hall LR, Morgan JS, O'Connor S, Marx PA, Meythaler M, Goldstein S, Buckler-White A, Kaur A, Hirsch VM, Johnson WE - PLoS Biol. (2010)

Differential restriction of SIVmac and SIVsm strains by multiple alleles of rhesus macaque TRIM5.Single-cycle infectivity was measured on a panel of cell lines stably expressing six common alleles of rhesus TRIM5 [15],[17]. Alleles tested included three TRIM5TFP (Mamu-1, Mamu-2, Mamu-3, dark blue bars), two TRIM5Q (Mamu-4 and Mamu-5, light blue bars), and TRIM5CypA (orange bars). Control cells stably expressing the empty vector served as a negative control (black bars). Of the four closely related SIVs, only the rhesus macaque isolate SIVmac239 is resistant to multiple alleles. Infectivity (% GFP-positive cells) was measured by flow-cytometry (error bars indicate ± SEM). Virus stocks were first titered by serial dilution on parental cells and normalized (Figure S1). Stable cell lines were generated from CRFK cells as described in the Materials and Methods section. HIV-1NL4-3 was used as a positive control to confirm expression and function of the TRIM5Q/Q alleles. (A) SIVmac239; (B) SIVsmE041; (C) SIVsmE543-3; (D) SIVstm37/13; (E) HIV-1NL4-3; (F) Immunoblot confirming expression of HA-tagged TRIM5 proteins in cell lysates. upper panel, TRIM5 proteins; lower panel, β-actin loading control. Lanes: (1) Mamu-1; (2) Mamu-2; (3) Mamu-3; (4) Mamu-4; (5) Mamu-5; M, protein standard; TC, TRIM5CypA; V, control cells expressing vector-only. GenBank accession number for the SIVsmE041 clone: HM059825.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2927514&req=5

pbio-1000462-g002: Differential restriction of SIVmac and SIVsm strains by multiple alleles of rhesus macaque TRIM5.Single-cycle infectivity was measured on a panel of cell lines stably expressing six common alleles of rhesus TRIM5 [15],[17]. Alleles tested included three TRIM5TFP (Mamu-1, Mamu-2, Mamu-3, dark blue bars), two TRIM5Q (Mamu-4 and Mamu-5, light blue bars), and TRIM5CypA (orange bars). Control cells stably expressing the empty vector served as a negative control (black bars). Of the four closely related SIVs, only the rhesus macaque isolate SIVmac239 is resistant to multiple alleles. Infectivity (% GFP-positive cells) was measured by flow-cytometry (error bars indicate ± SEM). Virus stocks were first titered by serial dilution on parental cells and normalized (Figure S1). Stable cell lines were generated from CRFK cells as described in the Materials and Methods section. HIV-1NL4-3 was used as a positive control to confirm expression and function of the TRIM5Q/Q alleles. (A) SIVmac239; (B) SIVsmE041; (C) SIVsmE543-3; (D) SIVstm37/13; (E) HIV-1NL4-3; (F) Immunoblot confirming expression of HA-tagged TRIM5 proteins in cell lysates. upper panel, TRIM5 proteins; lower panel, β-actin loading control. Lanes: (1) Mamu-1; (2) Mamu-2; (3) Mamu-3; (4) Mamu-4; (5) Mamu-5; M, protein standard; TC, TRIM5CypA; V, control cells expressing vector-only. GenBank accession number for the SIVsmE041 clone: HM059825.
Mentions: Infectivity of all four viruses was measured on cell lines stably expressing six common alleles of rhesus macaque TRIM5, including three TRIM5TFP alleles, two TRIM5Q alleles, and the TRIM5CypA allele (Figure 2). SIVmac239 was resistant to all six. SIVsmE041, SIVsmE543, and SIVstm were also resistant to TRIM5Q but unlike SIVmac239 were sensitive to both TRIM5CypA alleles and TRIM5TFP alleles (Figure 2B,C,D). Of the four SIV strains tested, only SIVmac239 is the product of decades of replication and spread in rhesus macaques. Thus, the comparison suggests that sensitivity to TRIM5TFP and TRIM5CypA alleles represents the ancestral phenotype and that emergence of SIVsm (as SIVmac) in rhesus macaques required acquisition of adaptive changes to overcome those particular types of alleles. In contrast, the SIVsm variants that first invaded rhesus macaques were probably inherently resistant to TRIM5Q alleles.

Bottom Line: Simian immunodeficiency viruses of sooty mangabeys (SIVsm) are the source of multiple, successful cross-species transmissions, having given rise to HIV-2 in humans, SIVmac in rhesus macaques, and SIVstm in stump-tailed macaques.Surprisingly, transmission occurred even in individuals bearing restrictive TRIM5 genotypes, resulting in attenuation of replication rather than an outright block to infection.In cell-culture assays, the same TRIM5 alleles associated with viral suppression in vivo blocked infectivity of two SIVsm strains, but not the macaque-adapted strain SIVmac239.

View Article: PubMed Central - PubMed

Affiliation: New England Primate Research Center, Department of Microbiology and Molecular Genetics, Harvard Medical School, Southborough, Massachusetts, United States of America.

ABSTRACT
Simian immunodeficiency viruses of sooty mangabeys (SIVsm) are the source of multiple, successful cross-species transmissions, having given rise to HIV-2 in humans, SIVmac in rhesus macaques, and SIVstm in stump-tailed macaques. Cellular assays and phylogenetic comparisons indirectly support a role for TRIM5alpha, the product of the TRIM5 gene, in suppressing interspecies transmission and emergence of retroviruses in nature. Here, we investigate the in vivo role of TRIM5 directly, focusing on transmission of primate immunodeficiency viruses between outbred primate hosts. Specifically, we retrospectively analyzed experimental cross-species transmission of SIVsm in two cohorts of rhesus macaques and found a significant effect of TRIM5 genotype on viral replication levels. The effect was especially pronounced in a cohort of animals infected with SIVsmE543-3, where TRIM5 genotype correlated with approximately 100-fold to 1,000-fold differences in viral replication levels. Surprisingly, transmission occurred even in individuals bearing restrictive TRIM5 genotypes, resulting in attenuation of replication rather than an outright block to infection. In cell-culture assays, the same TRIM5 alleles associated with viral suppression in vivo blocked infectivity of two SIVsm strains, but not the macaque-adapted strain SIVmac239. Adaptations appeared in the viral capsid in animals with restrictive TRIM5 genotypes, and similar adaptations coincide with emergence of SIVmac in captive macaques in the 1970s. Thus, host TRIM5 can suppress viral replication in vivo, exerting selective pressure during the initial stages of cross-species transmission.

Show MeSH
Related in: MedlinePlus