TABLE 1.

T-cell subset response to VV in immunized animalsa

VaccinationStimulationNo. of T cells (SEM)
CD8+CD8+/IFN-γ+CD4+CD4+/IFN-γ+
VV/NEA-20 (infVV)836.0 (± 87)282.0 (± 14)1,941.5 (± 47.5)271.4 (± 25)
A-20825.0 (± 157)40.5 (± 17.5)2,073.0 (± 170)75.3 (± 14)
cl7 (infVV)815.4 (± 91.2)212.6 (± 73)NANA
cl71,086.3 (± 113.5)73.0 (± 24)NANA
VV/scarA-20 (infVV)475.1 (± 43)135.6 (± 19.6)1,987.2 (± 244)303.5 (± 65.5)
A-20594.3 (± 17)52.0 (± 10.4)2,209.2 (± 365)81.3 (± 12.7)
cl7 (infVV)589.5 (± 33.5)173.0 (± 22)NANA
cl7679.0 (± 32.6)81.5 (± 12.5)NANA
Nonvaccinated controlsA-20 (infVV)644.0 (± 48.6)75.5 (± 15.5)1,934.0 (± 50)82.5 (± 9.6)
A-20667.5 (± 75.5)42.5 (± 14.8)1,767.0 (± 56)54.5 (± 15.4)
cl7 (infVV)518.5 (± 10.6)61.5 (± 12.1)NANA
cl7691.5 (± 22.1)44.0 (± 25.7)NANA
  • a Splenocytes were stimulated with A-20 cells infected with VVWR (infVV), with uninfected A-20 cells serving as a background control. VV-specific T cells expressing IFN-γ were detected by intracellular staining and enumerated by flow cytometry, and percentages of CD8+ and CD4+ cells that were IFN-γ+ are indicated. A-20 is a syngeneic H-2d haplotype B-cell lymphoma cell line expressing both MHC class I and MHC class II molecules, and cl7 is a syngeneic H-2d haplotype fibroblast line expressing only the MHC class I molecule. These were used as stimulator cells to show that different populations of cells could be stimulated based on CD4/CD8 expression. IFN-γ-producing CD4 and CD8 cells were specifically generated in response to VV stimulation in animals vaccinated with either scarification or i.n. NE administration.