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A molecular tweezer antagonizes seminal amyloids and HIV infection.

Lump E, Castellano LM, Meier C, Seeliger J, Erwin N, Sperlich B, Stürzel CM, Usmani S, Hammond RM, von Einem J, Gerold G, Kreppel F, Bravo-Rodriguez K, Pietschmann T, Holmes VM, Palesch D, Zirafi O, Weissman D, Sowislok A, Wettig B, Heid C, Kirchhoff F, Weil T, Klärner FG, Schrader T, Bitan G, Sanchez-Garcia E, Winter R, Shorter J, Münch J - Elife (2015)

Bottom Line: In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils.We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism.CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.

ABSTRACT
Semen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides that target viral components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement of viral infection by preventing the formation of virion-amyloid complexes and by directly disrupting the membrane integrity of HIV and other enveloped viruses. We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism. CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself. These combined anti-amyloid and antiviral activities make CLR01 a promising topical microbicide for blocking infection by HIV and other sexually transmitted viruses.

No MeSH data available.


Related in: MedlinePlus

CLR01 is a broad-spectrum inhibitor of enveloped viruses.(A) Human cytomegalovirus was incubated with PBS, 100 µM CLR03, or 100 µM CLR01. Afterwards, HFF cells were infected and immediate early (IE) antigen positive cells were counted 1 day post infection as a measure for infectivity. Values are means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (B) Herpes simplex virus type 2 comprising a GFP reporter gene was treated with PBS, 100 µM CLR03 or 100 µM CLR01 and added to Vero cells. GFP-positive cells were counted using flow cytometry 2 days post infection. Values represent means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (C) A luciferase encoding hepatitis C virus was treated with 150 µM CLR01 or 150 µM CLR03 and used for infection of Huh-7.5 reporter cells. Infection was measured 3 days post infection. Values represent means ±SEM (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (D) A GFP-reporter adenovirus type 5 was added to A549 cells after treatment with 158 µM CLR01 or 158 µM CLR03. GFP positive cells were counted using flow cytometry 1 day post infection. Values represent means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant).DOI:http://dx.doi.org/10.7554/eLife.05397.027
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fig8: CLR01 is a broad-spectrum inhibitor of enveloped viruses.(A) Human cytomegalovirus was incubated with PBS, 100 µM CLR03, or 100 µM CLR01. Afterwards, HFF cells were infected and immediate early (IE) antigen positive cells were counted 1 day post infection as a measure for infectivity. Values are means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (B) Herpes simplex virus type 2 comprising a GFP reporter gene was treated with PBS, 100 µM CLR03 or 100 µM CLR01 and added to Vero cells. GFP-positive cells were counted using flow cytometry 2 days post infection. Values represent means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (C) A luciferase encoding hepatitis C virus was treated with 150 µM CLR01 or 150 µM CLR03 and used for infection of Huh-7.5 reporter cells. Infection was measured 3 days post infection. Values represent means ±SEM (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (D) A GFP-reporter adenovirus type 5 was added to A549 cells after treatment with 158 µM CLR01 or 158 µM CLR03. GFP positive cells were counted using flow cytometry 1 day post infection. Values represent means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant).DOI:http://dx.doi.org/10.7554/eLife.05397.027

Mentions: CLR01 exhibited a surprising ability to disrupt viral membranes (Figures 5A, 6A,C–F), but did not affect cell viability like various non-ionic and anionic surfactants (Figure 4—figure supplement 3). We therefore explored whether CLR01 could act as a general inhibitor of enveloped viruses. To test this concept, human cytomegalovirus (HCMV), herpes simplex virus type 2 (HSV-2), and hepatitis C virus (HCV) were treated with CLR01 or CLR03 and then assessed for their ability to infect target cells. Remarkably, CLR01, but not CLR03, reduced infection rates of all three analyzed enveloped viruses (Figure 8A–C). By contrast, CLR01 did not inhibit infection by the non-enveloped human adenovirus type 5 (HAdV5) (Figure 8D). Thus, the tweezer is a broad-spectrum inhibitor of enveloped viruses, including viruses that can be sexually transmitted such as HIV-1, HSV-2, and HCV.10.7554/eLife.05397.027Figure 8.CLR01 is a broad-spectrum inhibitor of enveloped viruses.


A molecular tweezer antagonizes seminal amyloids and HIV infection.

Lump E, Castellano LM, Meier C, Seeliger J, Erwin N, Sperlich B, Stürzel CM, Usmani S, Hammond RM, von Einem J, Gerold G, Kreppel F, Bravo-Rodriguez K, Pietschmann T, Holmes VM, Palesch D, Zirafi O, Weissman D, Sowislok A, Wettig B, Heid C, Kirchhoff F, Weil T, Klärner FG, Schrader T, Bitan G, Sanchez-Garcia E, Winter R, Shorter J, Münch J - Elife (2015)

CLR01 is a broad-spectrum inhibitor of enveloped viruses.(A) Human cytomegalovirus was incubated with PBS, 100 µM CLR03, or 100 µM CLR01. Afterwards, HFF cells were infected and immediate early (IE) antigen positive cells were counted 1 day post infection as a measure for infectivity. Values are means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (B) Herpes simplex virus type 2 comprising a GFP reporter gene was treated with PBS, 100 µM CLR03 or 100 µM CLR01 and added to Vero cells. GFP-positive cells were counted using flow cytometry 2 days post infection. Values represent means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (C) A luciferase encoding hepatitis C virus was treated with 150 µM CLR01 or 150 µM CLR03 and used for infection of Huh-7.5 reporter cells. Infection was measured 3 days post infection. Values represent means ±SEM (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (D) A GFP-reporter adenovirus type 5 was added to A549 cells after treatment with 158 µM CLR01 or 158 µM CLR03. GFP positive cells were counted using flow cytometry 1 day post infection. Values represent means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant).DOI:http://dx.doi.org/10.7554/eLife.05397.027
© Copyright Policy
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC4536748&req=5

fig8: CLR01 is a broad-spectrum inhibitor of enveloped viruses.(A) Human cytomegalovirus was incubated with PBS, 100 µM CLR03, or 100 µM CLR01. Afterwards, HFF cells were infected and immediate early (IE) antigen positive cells were counted 1 day post infection as a measure for infectivity. Values are means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (B) Herpes simplex virus type 2 comprising a GFP reporter gene was treated with PBS, 100 µM CLR03 or 100 µM CLR01 and added to Vero cells. GFP-positive cells were counted using flow cytometry 2 days post infection. Values represent means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (C) A luciferase encoding hepatitis C virus was treated with 150 µM CLR01 or 150 µM CLR03 and used for infection of Huh-7.5 reporter cells. Infection was measured 3 days post infection. Values represent means ±SEM (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant; *** denotes p < 0.001). (D) A GFP-reporter adenovirus type 5 was added to A549 cells after treatment with 158 µM CLR01 or 158 µM CLR03. GFP positive cells were counted using flow cytometry 1 day post infection. Values represent means ±SD (n = 3). Unpaired t-tests were used to compare the buffer control to the CLR03 or CLR01 condition (ns denotes not significant).DOI:http://dx.doi.org/10.7554/eLife.05397.027
Mentions: CLR01 exhibited a surprising ability to disrupt viral membranes (Figures 5A, 6A,C–F), but did not affect cell viability like various non-ionic and anionic surfactants (Figure 4—figure supplement 3). We therefore explored whether CLR01 could act as a general inhibitor of enveloped viruses. To test this concept, human cytomegalovirus (HCMV), herpes simplex virus type 2 (HSV-2), and hepatitis C virus (HCV) were treated with CLR01 or CLR03 and then assessed for their ability to infect target cells. Remarkably, CLR01, but not CLR03, reduced infection rates of all three analyzed enveloped viruses (Figure 8A–C). By contrast, CLR01 did not inhibit infection by the non-enveloped human adenovirus type 5 (HAdV5) (Figure 8D). Thus, the tweezer is a broad-spectrum inhibitor of enveloped viruses, including viruses that can be sexually transmitted such as HIV-1, HSV-2, and HCV.10.7554/eLife.05397.027Figure 8.CLR01 is a broad-spectrum inhibitor of enveloped viruses.

Bottom Line: In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils.We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism.CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Virology, Ulm University Medical Center, Ulm, Germany.

ABSTRACT
Semen is the main vector for HIV transmission and contains amyloid fibrils that enhance viral infection. Available microbicides that target viral components have proven largely ineffective in preventing sexual virus transmission. In this study, we establish that CLR01, a 'molecular tweezer' specific for lysine and arginine residues, inhibits the formation of infectivity-enhancing seminal amyloids and remodels preformed fibrils. Moreover, CLR01 abrogates semen-mediated enhancement of viral infection by preventing the formation of virion-amyloid complexes and by directly disrupting the membrane integrity of HIV and other enveloped viruses. We establish that CLR01 acts by binding to the target lysine and arginine residues rather than by a non-specific, colloidal mechanism. CLR01 counteracts both host factors that may be important for HIV transmission and the pathogen itself. These combined anti-amyloid and antiviral activities make CLR01 a promising topical microbicide for blocking infection by HIV and other sexually transmitted viruses.

No MeSH data available.


Related in: MedlinePlus