TY - JOUR
T1 - AAV2-Mediated Subretinal Gene Transfer of hIFN-alpha Attenuates Experimental Autoimmune Uveoretinitis in Mice
AU - Tian, Lichun
AU - Yang, Peizeng
AU - Lei, Bo
AU - Shao, Ju
AU - Wang, Chaokui
AU - Xiang, Qin
AU - Wei, Lin
AU - Peng, Zhougui
AU - Kijlstra, Aize
PY - 2011/5/17
Y1 - 2011/5/17
N2 - Background: Recent reports show that gene therapy may provide a long-term, safe and effective intervention for human diseases. In this study, we investigated the effectiveness of adeno-associated virus 2 (AAV2) based human interferon-alpha (hIFN-alpha) gene therapy in experimental autoimmune uveoretinitis (EAU), a classic model for human uveitis. Methodology/Principal Findings: An AAV2 vector harboring the hIFN-alpha gene (AAV2.hIFN-alpha) was subretinally injected into B10RIII mice at two doses (1.5x10(6) vg, 1.5x10(8) vg). AAV2 vector encoding green fluorescent protein (AAV2.GFP) was used as a control (5x10(8) vg). The expression of hIFN-alpha in homogenized eyes and serum was detected by ELISA three weeks after injection. The biodistribution of vector DNA in the injected eyes, contralateral eyes and distant organs was determined by PCR. EAU was induced by immunization with IRBP(161-180) three weeks following vector injections, and evaluated clinically and pathologically. IRBP-specific proliferation and IL-17 expression of lymphocytes from the spleen and lymph nodes were assayed to test the influence of the subretinal delivery of AAV2. hIFN-alpha on the systemic immune response. hIFN-alpha was effectively expressed in the eyes from three weeks to three months following subretinal injection of AAV2. hIFN-alpha vector. DNA of AAV2.GFP was observed only in the injected eyes, but not in the distant organs or contralateral eyes. Subretinal injection of both doses significantly attenuated EAU activity clinically and histologically. For the lower dose, there was no difference concerning lymphocyte proliferation and IL-17 production among the AAV2.hIFN-alpha, AAV2.GFP and PBS injected mice. However, the higher dose of AAV2.hIFN-alpha significantly suppressed lymphocyte proliferation and IL-17 production. Conclusions/Significance: Subretinal delivery of AAV2.hIFN-alpha lead to an effective expression within the eye for at least three months and significantly attenuated EAU activity. AAV2.hIFN-alpha was shown to inhibit the systemic IRBP-specific immune response.
AB - Background: Recent reports show that gene therapy may provide a long-term, safe and effective intervention for human diseases. In this study, we investigated the effectiveness of adeno-associated virus 2 (AAV2) based human interferon-alpha (hIFN-alpha) gene therapy in experimental autoimmune uveoretinitis (EAU), a classic model for human uveitis. Methodology/Principal Findings: An AAV2 vector harboring the hIFN-alpha gene (AAV2.hIFN-alpha) was subretinally injected into B10RIII mice at two doses (1.5x10(6) vg, 1.5x10(8) vg). AAV2 vector encoding green fluorescent protein (AAV2.GFP) was used as a control (5x10(8) vg). The expression of hIFN-alpha in homogenized eyes and serum was detected by ELISA three weeks after injection. The biodistribution of vector DNA in the injected eyes, contralateral eyes and distant organs was determined by PCR. EAU was induced by immunization with IRBP(161-180) three weeks following vector injections, and evaluated clinically and pathologically. IRBP-specific proliferation and IL-17 expression of lymphocytes from the spleen and lymph nodes were assayed to test the influence of the subretinal delivery of AAV2. hIFN-alpha on the systemic immune response. hIFN-alpha was effectively expressed in the eyes from three weeks to three months following subretinal injection of AAV2. hIFN-alpha vector. DNA of AAV2.GFP was observed only in the injected eyes, but not in the distant organs or contralateral eyes. Subretinal injection of both doses significantly attenuated EAU activity clinically and histologically. For the lower dose, there was no difference concerning lymphocyte proliferation and IL-17 production among the AAV2.hIFN-alpha, AAV2.GFP and PBS injected mice. However, the higher dose of AAV2.hIFN-alpha significantly suppressed lymphocyte proliferation and IL-17 production. Conclusions/Significance: Subretinal delivery of AAV2.hIFN-alpha lead to an effective expression within the eye for at least three months and significantly attenuated EAU activity. AAV2.hIFN-alpha was shown to inhibit the systemic IRBP-specific immune response.
U2 - 10.1371/journal.pone.0019542
DO - 10.1371/journal.pone.0019542
M3 - Article
C2 - 21611186
SN - 1932-6203
VL - 6
SP - 8
JO - PLOS ONE
JF - PLOS ONE
IS - 5
ER -