Table 2 Contribution of factors to conclusion of ChR2-specific immunogenicity.
Potential Causes | TA histology | Spinal Cord IHC | Sciatic IF | Rag2−/− Electrophys. | Excitotox. Control | Immuno-suppression | CAG muscle histology | CAG mortality | No Reporter |
|---|---|---|---|---|---|---|---|---|---|
Phototoxicity | + | − | |||||||
Excitotoxicity | + | − | |||||||
ChR2 toxicity | + | − | |||||||
EYFP toxicity | + | − | |||||||
AAV immunogenicity | + | + | + | + | + | − | − | ||
ChR2 immunogenicity | + | + | + | + | + | + | + | ||
EYFP immunogenicity | + | + | + | + | + | + | − | ||
Episomal DNA loss | − | − | |||||||
Epigenetic silencing | − | − | |||||||
Transgene protein degradation or mRNA lysis | − | − | |||||||
Anatomical | − | − | − | ||||||
Rationale | Denervation atrophy suggests neuronal damage | CD8+ & CD68+ WBC home to ChR2+ Neurons | Axons healthy, with slight elevation of CTLs | 5/5 Rag2−/− compared to 1/5 WT with expression | WT rats low expression & pos. immune panel whether stim or not | Tacrolimus rats maintain expression & neg. immune panel | Inflammation only present in CAG-ChR2-EYFP+ myocytes | AAV is same, but significant mortality in CAG rats | Loss of expression & pos. immune panel without reporter |
