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. Author manuscript; available in PMC: 2011 Mar 17.
Published in final edited form as: Vaccine. 2008 Jun 2;26(32):4062–4072. doi: 10.1016/j.vaccine.2008.05.018

Figure 4.

Figure 4

CD40L expressed by recombinant ALVAC virus promotes human MDDCs maturation independent of TNF-α secretion. Immature MDDCs were infected with recombinant ALVAC virus expressing human CD40L, vA3131-2; an ALVAC-HIV vaccine, vCP205; or a parental control, ALVAC II at an MOI of 10, or incubated with medium alone at 37°C for 48 h. Expression of surface molecules was analyzed by flow cytometry. (A) Numbers in CD83 row represent percentage of CD83 positive cells in total MDDCs and numbers in CD86 and CD80 rows represent the mean fluorescence intensity of specific staining (histogram with solid line) subtracted from the value of background staining with matched isotype control mouse mAbs (dotted histogram). Data shown are from HIV-1-infected participant #1 and representative of six experiments performed with MDDCs obtained from three HIV-1-uninfected blood donors and three HIV-1-infected individuals. (B) Pooled data from all six participants studied are shown. Data shown are mean±SEM. **: P<0.01. (C) Immature human MDDCs were infected or not (medium) with vA3131-2 (CD40L expressing), vCP205, or ALVAC II at an MOI of 10 and then incubated in the presence of a blocking anti-human TNF-α Ab or the control isotype immunoglobulin (20 μg/mL). After 24 h incubation, MDDCs were collected and stained for CD83 expression to monitor DC maturation. Numbers in each dot plot represent percentage of CD83 positive cells in total MDDCs. Expression of CD83 on MDDCs treated with isotype immunoglobulin is similar to that on MDDCs treated with medium. Increasing the concentration of blocking anti-human TNF-α Ab up to 100 μg/mL did not further reduce the CD83 expression in all conditions (data not shown). The results represent one of five experiments.