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. 2004 Jun;112(2):328–338. doi: 10.1111/j.1365-2567.2004.01877.x

Figure 1.

Figure 1

Antigen-specific lymphocyte proliferation induced by binding of tgD or tgD-CD154 to bovine PBMCs. Supernatants from (a) mock-transfected, (b) pSLIAtgD-transfected, or (c) pSLIAtgD-CD154-transfected Cos-7 cells were incubated with bovine PBMCs. Bound protein was detected with a gD-specific mAb cocktail, followed by FITC-conjugated goat anti-mouse immunoglobulin. (d) PBMCs were isolated from naïve (tgD–) and tgD immunized (tgD+) cattle (n=3) and incubated for 30 min at 4° with either pSLIAtgD (tgD s/n pul) or pSLIAtgD-CD154 (tgD-CD154 s/n pul) transfected Cos-7 cell supernatants or with 3 μg/well of purified tgD (PtgD pul). Subsequently, PBMCs were washed twice and 2 × 105 cells/well were cultured for 72 hr. As a positive control, 2 × 105 PBMCs/well were also cultured for 72 hr with 3 μg of purified tgD (PtgD). Cell proliferation was detected by adding methyl-[3H]thymidine during the last 8 hr of culture. Data are shown as average c.p.m.+SEM for triplicate cultures.