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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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The expression of transgenes following subretinal injection. In the AAV2.IFN-α injected eyes, the level of IFN-α increases from 14 days (the first time point tested) to three months after injection. For the eyes receiving AAV2.IFN-α combined with AAV2.IL-4 injection, IFN-α level reaches a peak on day 42 and remains at a moderate level until three months after injection (a). IL-4 expression is similar in eyes receiving an injection of AAV2.IL-4 alone as compared to eyes receiving AAV2.IL-4 combined with AAV2.IFN-α (b). Results are expressed as the mean±standard deviation.
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pone-0037995-g002: The expression of transgenes following subretinal injection. In the AAV2.IFN-α injected eyes, the level of IFN-α increases from 14 days (the first time point tested) to three months after injection. For the eyes receiving AAV2.IFN-α combined with AAV2.IL-4 injection, IFN-α level reaches a peak on day 42 and remains at a moderate level until three months after injection (a). IL-4 expression is similar in eyes receiving an injection of AAV2.IL-4 alone as compared to eyes receiving AAV2.IL-4 combined with AAV2.IFN-α (b). Results are expressed as the mean±standard deviation.

Mentions: Mice were subretinally injected with AAV2.IFN-α (1.5×107 vg) alone, AAV2.IL-4 (3.55×107 vg) alone, and AAV2.IFN-α combined with AAV2.IL-4. Two weeks following subretinal injection, the IFN-α and IL-4 level in ocular fluid samples obtained from injected eyes was assayed by ELISA. Data showed that the expression of IFN-α and IL-4 was detectable two weeks following subretinal injection in each group. In the eye receiving an injection of AAV2.IFN-α (1.5×107 vg) alone, the level of IFN-α was 0.128 ng/ml on day 14, and 0.66 ng/ml on day 21 (Fig. 2a). The level increased further up to day 42 and started declining on day 90. Following the combined AAV2.IFN-α and AAV2.IL-4 vector injection, expression of IFN-α was observed on day 14, reached a peak on day 42 and remained detectable until three months after injection (Fig. 2a). For the expression of IL-4, the eyes injected with the AAV2.IL-4 vector alone and injected with the combined AAV2.IL-4 and AAV2.IFN-α showed a similar expression profile from 14 days to three months after subretinal injection (Fig. 2b). For all mice receiving subretinal injection of either AAV2.IFN-α alone, AAV2.IL-4 (3.55×107 vg) alone, or AAV2.IFN-α combined with AAV2.IL-4, IFN-α or IL-4 remained undetectable in undiluted serum and in the contralateral uninjected eyes over time (data not shown).


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)

The expression of transgenes following subretinal injection. In the AAV2.IFN-α injected eyes, the level of IFN-α increases from 14 days (the first time point tested) to three months after injection. For the eyes receiving AAV2.IFN-α combined with AAV2.IL-4 injection, IFN-α level reaches a peak on day 42 and remains at a moderate level until three months after injection (a). IL-4 expression is similar in eyes receiving an injection of AAV2.IL-4 alone as compared to eyes receiving AAV2.IL-4 combined with AAV2.IFN-α (b). Results are expressed as the mean±standard deviation.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0037995-g002: The expression of transgenes following subretinal injection. In the AAV2.IFN-α injected eyes, the level of IFN-α increases from 14 days (the first time point tested) to three months after injection. For the eyes receiving AAV2.IFN-α combined with AAV2.IL-4 injection, IFN-α level reaches a peak on day 42 and remains at a moderate level until three months after injection (a). IL-4 expression is similar in eyes receiving an injection of AAV2.IL-4 alone as compared to eyes receiving AAV2.IL-4 combined with AAV2.IFN-α (b). Results are expressed as the mean±standard deviation.
Mentions: Mice were subretinally injected with AAV2.IFN-α (1.5×107 vg) alone, AAV2.IL-4 (3.55×107 vg) alone, and AAV2.IFN-α combined with AAV2.IL-4. Two weeks following subretinal injection, the IFN-α and IL-4 level in ocular fluid samples obtained from injected eyes was assayed by ELISA. Data showed that the expression of IFN-α and IL-4 was detectable two weeks following subretinal injection in each group. In the eye receiving an injection of AAV2.IFN-α (1.5×107 vg) alone, the level of IFN-α was 0.128 ng/ml on day 14, and 0.66 ng/ml on day 21 (Fig. 2a). The level increased further up to day 42 and started declining on day 90. Following the combined AAV2.IFN-α and AAV2.IL-4 vector injection, expression of IFN-α was observed on day 14, reached a peak on day 42 and remained detectable until three months after injection (Fig. 2a). For the expression of IL-4, the eyes injected with the AAV2.IL-4 vector alone and injected with the combined AAV2.IL-4 and AAV2.IFN-α showed a similar expression profile from 14 days to three months after subretinal injection (Fig. 2b). For all mice receiving subretinal injection of either AAV2.IFN-α alone, AAV2.IL-4 (3.55×107 vg) alone, or AAV2.IFN-α combined with AAV2.IL-4, IFN-α or IL-4 remained undetectable in undiluted serum and in the contralateral uninjected eyes over time (data not shown).

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