The degree of adeno-associated virus-induced retinal inflammation varies based on serotype and route of delivery: intravitreal, subretinal, or suprachoroidal
Open accessAbstract
Adeno-associated virus (AAV)-mediated gene therapy has great potential for treating a wide range of retinal degenerative diseases. However, some initial enthusiasm for gene therapy has been tempered by emerging evidence of AAV-associated inflammation, which in several instances has contributed to clinical trial discontinuation. Currently, there is a paucity of data describing the variable immune responses to different AAV serotypes, and similarly, little is known regarding how these responses differ depending on route of ocular delivery, including in animal models of disease. In this study, we characterize the severity and retinal distribution of AAV-associated inflammation in rats triggered by delivery of five different AAV vectors (AAV1, AAV2, AAV6, AAV8, and AAV9), each of which contained enhanced green fluorescent protein (eGFP) driven under control of the constitutively active cytomegalovirus promoter. We further compare the inflammation across three different potential routes (intravitreal, subretinal, and suprachoroidal) of ocular delivery. Compared to buffer-injected controls for each route of delivery, AAV2 and AAV6 induced the most inflammation across all routes of delivery of vectors tested, with AAV6 inducing the highest levels of inflammation when delivered suprachoroidally. AAV1-induced inflammation was highest when delivered suprachoroidally, whereas minimal inflammation was seen with intravitreal delivery. In addition, AAV1, AAV2, and AAV6 each induce infiltration of adaptive immune cells like T cells and B cells into the neural retina, suggesting an innate adaptive response to a single dose of virus. AAV8 and AAV9 induced minimal inflammation across all routes of delivery. Importantly, the degree of inflammation was not correlated with vector-mediated transduction and expression of eGFP. These data emphasize the importance of considering ocular inflammation when selecting AAV serotypes and ocular delivery routes for the development of gene therapy strategies.
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In mice, IVT AAV2 injections without immunomodulation predominantly cause mild to moderate inflammation with ocular infiltration of CD45+ peripheral immune cells, including T cells.
“Preclinical investigations utilizing mice have limited utility in predicting severe adverse events, as outcomes from AAV2 IVT injections without immunomodulation predominantly reveal mild to moderate inflammation accompanied by ocular infiltration of CD45+ peripheral immune cells, including T cells (30, 32–34).”
From Intravitreal AAV vector delivery induces integrin-dependent ocular inflammation, complement activation, and antiviral and DNA damage responses (2026) · cites this paper as [34]
The Delivery of viral vector section (page unavailable) identifies the study animals as rats, not mice. The severity-and-location results (page unavailable) report inflammation after intravitreal AAV2 and CD3-positive T cells after AAV2 by all delivery routes. They do not establish predominantly mild-to-moderate inflammation or identify infiltrating cells as CD45-positive; the Discussion describes their origin as plausible rather than confirmed. (LLM, conf=0.88)
Experimental — accuracy not yet measuredMouse preclinical investigations have limited utility in predicting severe adverse events from IVT AAV.
“Preclinical investigations utilizing mice have limited utility in predicting severe adverse events”
From Intravitreal AAV vector delivery induces integrin-dependent ocular inflammation, complement activation, and antiviral and DNA damage responses (2026) · cites this paper as [34]
The Discussion (page unavailable) says species-specific differences make it difficult to translate animal-model findings about adverse effects to humans. It also notes that rats, rather than murine models, are the authors’ preferred model for preclinical toxicity testing. The paper does not specifically establish that mouse studies have limited utility for predicting severe adverse events from intravitreal AAV. (LLM, conf=0.84)
Experimental — accuracy not yet measuredThe type of promoter, AAV serotype and type of transgene affect treatment outcome and the degree of post-operative inflammation.
“Finally, the type of promoter [14], AAV serotype [15] and type of transgene [16] also affect treatment outcome and the degree of post-operative inflammation.”
From Characterization of anti-AAV2 neutralizing antibody levels in sheep prior to and following intravitreal AAV2.7m8 injection (2024) · cites this paper as [15]
The discussion (page unavailable) reports that AAV serotype affects retinal inflammation in rats, and notes differences in retinal transduction between serotypes. It does not establish that serotype affects clinical treatment outcomes; the authors say they did not investigate consequences for retinal cell survival or death. The promoter and transgene parts of the sentence have separate citations and are not judged here. (LLM, conf=0.85)
Experimental — accuracy not yet measuredIba1+ cells from AAV-injected pigs showed upregulated SLA-DR, consistent with microglia activation.
“Moreover, Iba1+ cells from injected animals showed upregulated Swine Leukocyte Antigen (SLA) major histocompatibility complex (MHC) class II DR molecules (SLA-DR) (Figures 5D, F, G), consistent with microglia activation and in line with previous studies of subretinal and IVT AAV delivery in animal models (13, 34).”
From Intravitreal AAV vector delivery induces integrin-dependent ocular inflammation, complement activation, and antiviral and DNA damage responses (2026) · cites this paper as [34]
Early manifestations of GTAU may be subclinical based on animal studies demonstrating microglial activation and immune cell infiltration in the outer retina without overt clinical signs of intraocular inflammation.
“Early manifestations of GTAU may be subclinical based on animal studies demonstrating microglial activation and immune cell infiltration in the outer retina without overt clinical signs of intraocular inflam mation (Chan et al., 2021b; Chandler et al., 2021; Wiley et al., 2023).”
From Gene Therapy-Associated Uveitis (GTAU): Understanding and mitigating the adverse immune response in retinal gene therapy (2025) · cites this paper as Wiley et al., 2023
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Cites
Russell, S., Bennett, J., Wellman, J. A., Chung, D. C., Yu, Z.-F., Tillman, A., Wittes, J., Pappas, J., Elci, O., McCague, S., et al. (2017) · The Lancet · doi:10.1016/s0140-6736(17)31868-8 · Cited by 1,952 · see citing works
Self-citationBennett, J., Wellman, J., Marshall, K. A., McCague, S., Ashtari, M., DiStefano-Pappas, J., Elci, O. U., Chung, D. C., Sun, J., Wright, J. F., et al. (2016) · The Lancet · doi:10.1016/s0140-6736(16)30371-3 · Cited by 461 · see citing works
Microglial phagocytosis of living photoreceptors contributes to inherited retinal degeneration
Zhao, L., Zabel, M. K., Wang, X., Ma, W., Shah, P., Fariss, R. N., Qian, H., Parkhurst, C. N., Gan, W., & Wong, W. T. (2015) · EMBO Molecular Medicine · doi:10.15252/emmm.201505298 · Cited by 451 · see citing works
O’Koren, E. G., Yu, C., Klingeborn, M., Wong, A. Y. W., Prigge, C. L., Mathew, R., Kalnitsky, J., Msallam, R. A., Silvin, A., Kay, J. N., et al. (2019) · Immunity · doi:10.1016/j.immuni.2019.02.007 · Cited by 440 · see citing works
Cukras, C., Wiley, H. E., Jeffrey, B. G., Sen, H. N., Turriff, A., Zeng, Y., Vijayasarathy, C., Marangoni, D., Ziccardi, L., Kjellstrom, S., et al. (2018) · Molecular Therapy · doi:10.1016/j.ymthe.2018.05.025 · Cited by 268 · cited 3× · see citing works
“6,7,23 By evaluating serotype and ocular delivery route, this study primarily demonstrates that exposure to different viral components ( e.g. , viral coats, asvariesbyserotype)andocular tissue exposure (asvariesby route of delivery) result in different inflammatory responses.”
Bucher, K., Rodríguez-Bocanegra, E., Dauletbekov, D., & Fischer, M. D. (2021) · Progress in Retinal and Eye Research · doi:10.1016/j.preteyeres.2020.100915 · Cited by 242 · see citing works
Engineering adeno-associated viral vectors to evade innate immune and inflammatory responses
Chan, Y. K., Wang, S. K., Chu, C. J., Copland, D. A., Letizia, A. J., Costa Verdera, H., Chiang, J. J., Sethi, M., Wang, M. K., Neidermyer, W. J., et al. (2021) · Science Translational Medicine · doi:10.1126/scitranslmed.abd3438 · Cited by 215 · see citing works
Zabel, M. K., Zhao, L., Zhang, Y., Gonzalez, S. R., Ma, W., Wang, X., Fariss, R. N., & Wong, W. T. (2016) · Glia · doi:10.1002/glia.23016 · Cited by 210 · see citing works
Wang, M., Wang, X., Zhao, L., Ma, W., Rodríguez, I. R., Fariss, R. N., & Wong, W. T. (2014) · Journal of Neuroscience · doi:10.1523/jneurosci.3153-13.2014 · Cited by 201 · see citing works
Durability of Voretigene Neparvovec for Biallelic RPE65-Mediated Inherited Retinal Disease
Maguire, A. M., Russell, S., Chung, D. C., Yu, Z.-F., Tillman, A., Drack, A. V., Simonelli, F., Leroy, B. P., Reape, K. Z., High, K. A., et al. (2021) · Ophthalmology · doi:10.1016/j.ophtha.2021.03.031 · Cited by 194 · see citing works
(2008) · PubMed Central · Cited by 183 · see citing works
Two-Year Results After AAV2-Mediated Gene Therapy for Choroideremia: The Alberta Experience
Dimopoulos, I. S., Hoang, S. C., Radziwon, A., Binczyk, N. M., Seabra, M. C., MacLaren, R. E., Somani, R., Tennant, M. T. S., & MacDonald, I. M. (2018) · American Journal of Ophthalmology · doi:10.1016/j.ajo.2018.06.011 · Cited by 175 · see citing works
Garafalo, A. V., Cideciyan, A. V., Héon, E., Sheplock, R., Pearson, A., WeiYang Yu, C., Sumaroka, A., Aguirre, G. D., & Jacobson, S. G. (2020) · Progress in Retinal and Eye Research · doi:10.1016/j.preteyeres.2019.100827 · Cited by 173 · see citing works
Gange, W. S., Sisk, R. A., Besirli, C. G., Lee, T. C., Havunjian, M., Schwartz, H., Borchert, M., Sengillo, J. D., Mendoza‐Santiesteban, C. E., Berrocal, A. M., et al. (2021) · Ophthalmology Retina · doi:10.1016/j.oret.2021.03.016 · Cited by 157 · see citing works
AAV8 Can Induce Innate and Adaptive Immune Response in the Primate Eye
Reichel, F. F., Dauletbekov, D. L., Klein, R., Peters, T., Ochakovski, G. A., Seitz, I. P., Wilhelm, B., Ueffing, M., Biel, M., Wissinger, B., et al. (2017) · Molecular Therapy · doi:10.1016/j.ymthe.2017.08.018 · Cited by 145 · see citing works
Bouquet, C., Vignal Clermont, C., Galy, A., Fitoussi, S., Blouin, L., Munk, M. R., Valero, S., Meunier, S., Katz, B., Sahel, J. A., et al. (2019) · JAMA Ophthalmology · doi:10.1001/jamaophthalmol.2018.6902 · Cited by 102 · see citing works
Ma, W., Zhang, Y., Gao, C., Fariss, R. N., Tam, J., & Wong, W. T. (2017) · Scientific Reports · doi:10.1038/s41598-017-08702-7 · Cited by 102 · see citing works
de Guimarães, T. A. C., Georgiou, M., Bainbridge, J. W., & Michaelides, M. (2020) · British Journal of Ophthalmology · doi:10.1136/bjophthalmol-2020-316195 · Cited by 92 · see citing works
Rutar, M., Provis, J. M., & Valter, K. (2010) · Current Eye Research · doi:10.3109/02713681003682925 · Cited by 86 · see citing works
Accumulation of murine subretinal macrophages: effects of age, pigmentation and CX3CR1
Chinnery, H. R., McLenachan, S., Humphries, T. G., Kezic, J. M., Chen, X., Ruitenberg, M. J., & McMenamin, P. G. (2011) · Neurobiology of Aging · doi:10.1016/j.neurobiolaging.2011.03.010 · Cited by 78 · see citing works
Karlen, S. J., Miller, E. B., Wang, X., Levine, E. S., Zawadzki, R. J., & Burns, M. E. (2018) · Journal of Neuroinflammation · doi:10.1186/s12974-018-1365-4 · Cited by 77 · see citing works
Pennesi, M. E., Yang, P., Birch, D. G., Weng, C. Y., Moore, A. T., Iannaccone, A., Comander, J. I., Jayasundera, T., Chulay, J., Chulay, J., et al. (2022) · Ophthalmology Retina · doi:10.1016/j.oret.2022.06.013 · Cited by 71 · see citing works
Assessment of Adeno-Associated Virus Serotype Tropism in Human Retinal Explants
Wiley, L. A., Burnight, E. R., Kaalberg, E. E., Jiao, C., Riker, M. J., Halder, J. A., Luse, M. A., Han, I. C., Russell, S. R., Sohn, E. H., et al. (2017) · Human Gene Therapy · doi:10.1089/hum.2017.179 · Cited by 67 · see citing works
Self-citationBoswell, C. A., Mundo, E. E., Ulufatu, S., et al. (2014) · Molecular Pharmaceutics · doi:10.1021/mp400748t · Cited by 66 · see citing works
Han, I. C., Cheng, J. L., Burnight, E. R., Ralston, C. L., Fick, J. L., Thomsen, G. J., Tovar, E. F., Russell, S. R., Sohn, E. H., Mullins, R. F., et al. (2020) · Human Gene Therapy · doi:10.1089/hum.2020.043 · Cited by 60 · see citing works
Self-citationHost immune responses after suprachoroidal delivery of AAV8 in nonhuman primate eyes
Chung, S. H., Mollhoff, I. N., Mishra, A., Sin, T.-N., Ngo, T., Ciulla, T., Sieving, P., Thomasy, S. M., & Yiu, G. (2021) · Human Gene Therapy · doi:10.1089/hum.2020.281 · Cited by 57 · see citing works
Mullins, R. F., Kuehn, M. H., Radu, R. A., Enriquez, G., East, J. S., Schindler, E. I., Travis, G. H., & Stone, E. M. (2012) · Investigative Ophthalmology & Visual Science · doi:10.1167/iovs.12-9477 · Cited by 56 · see citing works
Self-citationKessel, L., Christensen, U. C., & Klemp, K. (2022) · Ophthalmology · doi:10.1016/j.ophtha.2022.06.018 · Cited by 46 · see citing works
Immune function in X-linked retinoschisis subjects in an AAV8-RS1 phase I/IIa gene therapy trial
Mishra, A., Vijayasarathy, C., Cukras, C. A., Wiley, H. E., Sen, H. N., Zeng, Y., Wei, L. L., & Sieving, P. A. (2021) · Molecular Therapy · doi:10.1016/j.ymthe.2021.02.013 · Cited by 38 · see citing works
Yang, J. M., Chung, S., Yun, K., et al. (2021) · Experimental & Molecular Medicine · doi:10.1038/s12276-021-00588-w · Cited by 35 · see citing works
Ocular Inflammation and Treatment Emergent Adverse Events in Retinal Gene Therapy
Mehta, N., Robbins, D. A., & Yiu, G. (2021) · International Ophthalmology Clinics · doi:10.1097/iio.0000000000000366 · Cited by 33 · see citing works
ICG-mediated photodisruption of the inner limiting membrane enhances retinal drug delivery
Peynshaert, K., Vanluchene, H., Clerck, K. D., Minnaert, A., Verhoeven, M., Gouspillou, N., Bostan, N., Hisatomi, T., Accou, G., Sauvage, F., et al. (2022) · Journal of Controlled Release · doi:10.1016/j.jconrel.2022.07.002 · Cited by 33 · see citing works
Han, I. C., Burnight, E. R., Ulferts, M. J., Worthington, K. S., Russell, S. R., Sohn, E. H., Mullins, R. F., Stone, E. M., Tucker, B. A., & Wiley, L. A. (2019) · Human Gene Therapy · doi:10.1089/hum.2019.159 · Cited by 30 · see citing works
Self-citationLópez, J., Borchert, M., Lee, T. C., & Nagiel, A. (2022) · British Journal of Ophthalmology · doi:10.1136/bjo-2022-321488 · Cited by 23 · see citing works
Han, I. C., Wiley, L. A., Ochoa, D., Lang, M. J., Harman, B. E., Sheehan, K. M., Mullins, R. F., Stone, E. M., & Tucker, B. A. (2022) · Gene Therapy · doi:10.1038/s41434-022-00365-y · Cited by 15 · see citing works
Self-citationTucker, B. A., Burnight, E. R., Cranston, C. M., Ulferts, M. J., Luse, M. A., Westfall, T. A., Scott, C. A., Marsden, A. N., Gibson-Corley, K. N., Wiley, L. A., et al. (2021) · Gene Therapy · doi:10.1038/s41434-021-00291-5 · Cited by 8 · see citing works
Self-citationHan, I. C., Burnight, E. R., Kaalberg, E. E., Boyce, T. M., Stone, E. M., Fingert, J. H., Mullins, R. F., Tucker, B. A., & Wiley, L. A. (2021) · Journal of Ocular Pharmacology and Therapeutics · doi:10.1089/jop.2021.0057 · Cited by 7 · see citing works
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Purdy, R., John, M., Bray, A., Clare, A. J., Copland, D. A., Chan, Y. K., Henderson, R. H., Nerinckx, F., Leroy, B. P., Yang, P., et al. (2025) · Progress in Retinal and Eye Research · doi:10.1016/j.preteyeres.2025.101354 · Cited by 24 · see citing works
“Early manifestations of GTAU may be subclinical based on animal studies demonstrating microglial activation and immune cell infiltration in the outer retina without overt clinical signs of intraocular inflammation (Chan et al., 2021b; Chandler et al., 2021; Wiley et al., 2023).”
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Clare, A. J., Langer, P. M., Ward, A., Chan, Y. K., Dick, A. D., & Copland, D. A. (2025) · Molecular Therapy · doi:10.1016/j.ymthe.2025.01.028 · Cited by 19 · see citing works
Ross, M., Sade, K., Obolensky, A., Averbukh, E., Desrosiers, M., Rosov, A., Dvir, H., Gootwine, E., Banin, E., Dalkara, D., et al. (2024) · Gene Therapy · doi:10.1038/s41434-024-00495-5 · Cited by 3 · see citing works
Costa-Verdera, H., Jamwal, D., Fabyanic, E., Hippen, A., Salvatore, M., Du, A., Majowicz, A., Markusic, D., Chatterton, J. E., Haurigot, V., et al. (2026) · Frontiers in Immunology · doi:10.3389/fimmu.2026.1799327 · Cited by 2 · see citing works
- Authors
- Wiley, L. A., Boyce, T. M., Meyering, E. E., Ochoa, D., Sheehan, K. M., Stone, E. M., Mullins, R. F., Tucker, B. A., & Han, I. C.
- Affiliations
- University of Iowa (US)
- Corresponding author
- Han, Ian C.
- Venue
- Human Gene Therapy
- Publisher
- SAGE Publications
- Published
- 2023-06
- Volume
- 34
- Issue
- 11-12
- Pages
- 530-539
- ISSN
- 1043-0342 (ISSN-L)
- 1557-7422
- PMC
- PMC10282814
- OpenAlex
- W4321004399
- Cited by
- 30
- References
- 37
- Citations by year
- 2026: 11 · 2025: 11 · 2024: 6 · 2023: 2
- FWCI
- 4.98
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Human Gene Therapy (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Eye Institute (P30EY025580)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- adeno-associated virus, AAV serotypes, retinal inflammation, ocular gene therapy, intravitreal delivery, subretinal delivery, AAV2, AAV6, AAV8, AAV9, AAV1, T cells, B cells, retinal degeneration
- MeSH terms
- Dependovirus, Animals, Genetic Vectors, Inflammation, Retina, Retinal Degeneration, Transduction, Genetic, Rats, Serogroup
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