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AAV2-mediated combined subretinal delivery of IFN-α and IL-4 reduces the severity of experimental autoimmune uveoretinitis.

Tian L, Lei B, Shao J, Wei L, Kijlstra A, Yang P - PLoS ONE (2012)

Bottom Line: IFN-α and IL-4 were effectively expressed in the eyes from three weeks to three months following subretinal injection of AAV2 vectors either alone or following combined administration and significantly attenuated EAU activity clinically and histopathologically.AAV2.IL-4 showed a better therapeutic effect as compared to AAV2.IFN-α.The combination of AAV2.IL-4 and AAV2.IFN-α was not significantly different as compared to AAV2.IL-4 alone.

View Article: PubMed Central - PubMed

Affiliation: The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology and Chongqing Eye Institute, Chongqing, People's Republic of China.

ABSTRACT
We previously showed that adeno-associated virus 2 (AAV2) mediated subretinal delivery of human interferon-alpha (IFN-α) could effectively inhibit experimental autoimmune uveoretinitis (EAU). In this study we investigated whether subretinal injection of both AVV2.IFN-α and AAV2.IL-4 had a stronger inhibition on EAU activity. B10RIII mice were subretinally injected with AAV2.IFN-α alone (1.5×10(7) vg), AAV2.IL-4 alone (3.55×10(7) vg), and AAV2.IFN-α combined with AAV2.IL-4. PBS, AAV2 vector encoding green fluorescent protein (AAV2.GFP) (5×10(7) vg) was subretinally injected as a control. IFN-α and IL-4 were effectively expressed in the eyes from three weeks to three months following subretinal injection of AAV2 vectors either alone or following combined administration and significantly attenuated EAU activity clinically and histopathologically. AAV2.IL-4 showed a better therapeutic effect as compared to AAV2.IFN-α. The combination of AAV2.IL-4 and AAV2.IFN-α was not significantly different as compared to AAV2.IL-4 alone. There was no difference concerning DTH (delayed-type hypersensitivity) reaction, lymphocyte proliferation and IL-17 production among the investigated treatment groups, suggesting that local retinal gene delivery did not affect the systemic immune response.

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Clinical evaluation of EAU activity.EAU was induced in groups of mice receiving subretinal vector or PBS injection. Severe uveitis is observed in the PBS (a) and AAV2.GFP injected eyes (b) as compared to the AAV vectors treated eyes (c–e). Three AAV vector treated groups show significantly attenuated EAU over time as compared with controls (f). Data are presented as mean±standard deviation. The clinical score shows that compared with controls, the AAV2.IFN-α treated group (p = 0.019, Mann-Whitney U test), the AAV2.IL-4 treated group (p<0.0001) and the combined treated group (p<0.0001) developed a significantly reduced EAU (g). Each point represents an individual eye. The average scores of each group are denoted by the horizontal bars.
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pone-0037995-g003: Clinical evaluation of EAU activity.EAU was induced in groups of mice receiving subretinal vector or PBS injection. Severe uveitis is observed in the PBS (a) and AAV2.GFP injected eyes (b) as compared to the AAV vectors treated eyes (c–e). Three AAV vector treated groups show significantly attenuated EAU over time as compared with controls (f). Data are presented as mean±standard deviation. The clinical score shows that compared with controls, the AAV2.IFN-α treated group (p = 0.019, Mann-Whitney U test), the AAV2.IL-4 treated group (p<0.0001) and the combined treated group (p<0.0001) developed a significantly reduced EAU (g). Each point represents an individual eye. The average scores of each group are denoted by the horizontal bars.

Mentions: Clinical signs were monitored after immunization by slit lamp microscopy. In PBS or AAV2.GFP injected eyes, a severe inflammatory reaction including conjunctival hyperemia, ciliary injection, corneal edema, aqueous cells and posterior synechiae was observed (Figs. 3a, b). A mild uveitis as manifested by conjunctival hyperemia or ciliary injection was observed in AAV2.IFN-α alone treated, AAV2.IL-4 alone treated, AAV2.IFN-α and AAV2.IL-4 combined treated eyes (Figs. 3c–e). Severity of EAU was clinically scored on a scale from 0 to 5. A significantly decreased activity of EAU throughout the course of the disease was observed in the three AAV vector treated groups as compared with the PBS or AAV2.GFP injected controls (Fig. 3f). Clinical scoring on day 12 following EAU induction showed that the severity of EAU in AAV2.IL-4 alone injected eyes and AAV2.IL-4 combined with AAV2.IFN-α treated eyes was significantly decreased when compared with PBS and AAV2.GFP injected controls (p<0.0001). Eyes receiving an injection of AAV2.IFN-α alone also showed a significantly decreased inflammation compared with the PBS (p = 0.001) or AAV2.GFP group (p = 0.013). AAV2.IL-4 treatment showed a much more dramatically increased blockade of EAU than AAV2.IFN-α treatment (p = 0.001). Subretinal injection of both AAV2.IL-4 and AAV2.IFN-α showed a stronger inhibitory effect on EAU activity as compared with AAV2.IFN-α administration alone (p<0.0001) and a trend of increased inhibition when compared with AAV2.IL-4 alone, although the difference did not reach statistical significance. (Fig. 3g).


AAV2-mediated combined subretinal delivery of IFN-α and IL-4 reduces the severity of experimental autoimmune uveoretinitis.

Tian L, Lei B, Shao J, Wei L, Kijlstra A, Yang P - PLoS ONE (2012)

Clinical evaluation of EAU activity.EAU was induced in groups of mice receiving subretinal vector or PBS injection. Severe uveitis is observed in the PBS (a) and AAV2.GFP injected eyes (b) as compared to the AAV vectors treated eyes (c–e). Three AAV vector treated groups show significantly attenuated EAU over time as compared with controls (f). Data are presented as mean±standard deviation. The clinical score shows that compared with controls, the AAV2.IFN-α treated group (p = 0.019, Mann-Whitney U test), the AAV2.IL-4 treated group (p<0.0001) and the combined treated group (p<0.0001) developed a significantly reduced EAU (g). Each point represents an individual eye. The average scores of each group are denoted by the horizontal bars.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3369876&req=5

pone-0037995-g003: Clinical evaluation of EAU activity.EAU was induced in groups of mice receiving subretinal vector or PBS injection. Severe uveitis is observed in the PBS (a) and AAV2.GFP injected eyes (b) as compared to the AAV vectors treated eyes (c–e). Three AAV vector treated groups show significantly attenuated EAU over time as compared with controls (f). Data are presented as mean±standard deviation. The clinical score shows that compared with controls, the AAV2.IFN-α treated group (p = 0.019, Mann-Whitney U test), the AAV2.IL-4 treated group (p<0.0001) and the combined treated group (p<0.0001) developed a significantly reduced EAU (g). Each point represents an individual eye. The average scores of each group are denoted by the horizontal bars.
Mentions: Clinical signs were monitored after immunization by slit lamp microscopy. In PBS or AAV2.GFP injected eyes, a severe inflammatory reaction including conjunctival hyperemia, ciliary injection, corneal edema, aqueous cells and posterior synechiae was observed (Figs. 3a, b). A mild uveitis as manifested by conjunctival hyperemia or ciliary injection was observed in AAV2.IFN-α alone treated, AAV2.IL-4 alone treated, AAV2.IFN-α and AAV2.IL-4 combined treated eyes (Figs. 3c–e). Severity of EAU was clinically scored on a scale from 0 to 5. A significantly decreased activity of EAU throughout the course of the disease was observed in the three AAV vector treated groups as compared with the PBS or AAV2.GFP injected controls (Fig. 3f). Clinical scoring on day 12 following EAU induction showed that the severity of EAU in AAV2.IL-4 alone injected eyes and AAV2.IL-4 combined with AAV2.IFN-α treated eyes was significantly decreased when compared with PBS and AAV2.GFP injected controls (p<0.0001). Eyes receiving an injection of AAV2.IFN-α alone also showed a significantly decreased inflammation compared with the PBS (p = 0.001) or AAV2.GFP group (p = 0.013). AAV2.IL-4 treatment showed a much more dramatically increased blockade of EAU than AAV2.IFN-α treatment (p = 0.001). Subretinal injection of both AAV2.IL-4 and AAV2.IFN-α showed a stronger inhibitory effect on EAU activity as compared with AAV2.IFN-α administration alone (p<0.0001) and a trend of increased inhibition when compared with AAV2.IL-4 alone, although the difference did not reach statistical significance. (Fig. 3g).

Bottom Line: IFN-α and IL-4 were effectively expressed in the eyes from three weeks to three months following subretinal injection of AAV2 vectors either alone or following combined administration and significantly attenuated EAU activity clinically and histopathologically.AAV2.IL-4 showed a better therapeutic effect as compared to AAV2.IFN-α.The combination of AAV2.IL-4 and AAV2.IFN-α was not significantly different as compared to AAV2.IL-4 alone.

View Article: PubMed Central - PubMed

Affiliation: The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology and Chongqing Eye Institute, Chongqing, People's Republic of China.

ABSTRACT
We previously showed that adeno-associated virus 2 (AAV2) mediated subretinal delivery of human interferon-alpha (IFN-α) could effectively inhibit experimental autoimmune uveoretinitis (EAU). In this study we investigated whether subretinal injection of both AVV2.IFN-α and AAV2.IL-4 had a stronger inhibition on EAU activity. B10RIII mice were subretinally injected with AAV2.IFN-α alone (1.5×10(7) vg), AAV2.IL-4 alone (3.55×10(7) vg), and AAV2.IFN-α combined with AAV2.IL-4. PBS, AAV2 vector encoding green fluorescent protein (AAV2.GFP) (5×10(7) vg) was subretinally injected as a control. IFN-α and IL-4 were effectively expressed in the eyes from three weeks to three months following subretinal injection of AAV2 vectors either alone or following combined administration and significantly attenuated EAU activity clinically and histopathologically. AAV2.IL-4 showed a better therapeutic effect as compared to AAV2.IFN-α. The combination of AAV2.IL-4 and AAV2.IFN-α was not significantly different as compared to AAV2.IL-4 alone. There was no difference concerning DTH (delayed-type hypersensitivity) reaction, lymphocyte proliferation and IL-17 production among the investigated treatment groups, suggesting that local retinal gene delivery did not affect the systemic immune response.

Show MeSH
Related in: MedlinePlus