Quantitation of OVA-specific cytokine responses in DO11.10 TCR transgenic BALB/c mice by ELISPOT. Mice were naive controls or had been injected intramuscularly with AAV-CMV-OVA vector (3 × 10¹² vg/animal). Panels A, C, E, and G show numbers of IFN-γ–secreting cells as spot-forming units (SFUs) per 10⁶ cells after mock or OVA stimulation. Panels B, D, F, and H show numbers of IL-4–secreting cells as SFUs per 10⁶ cells after mock or OVA stimulation. (A-B) Splenocytes, day 10 after gene transfer. (C-D) LN cells, day 10. (E-F) Splenocytes, day 30. (G-H) LN cells, day 30 after gene transfer. LNs were inguinal and popliteal LNs. D-LN indicates draining LNs of vector-injected leg muscle; ND-LN, nondraining LNs of noninjected contralateral leg. Results from D-LNs are marked by vertical arrow. All ELISPOT cultures were set up in quadruplicate. Spleens from each animal were processed and analyzed individually. LN cells were pooled for experimental groups (5 animals/group). Results are average of quadruplicate measurement ± SD.