Fig 4. Schematic representation of T cell enrichment, CFSE staining and cell transference procedure.

Groups of BALB/c mice (n = 13) received two doses of pcTPANS1 by the intramuscular (i.m.) route with two weeks of interval between each dose. Two weeks after the second dose, spleen and anti-NS1 antiserum samples were collected from these animals. T cell-enriched suspensions were obtained from isolated splenocytes by nylon wool purification and then labeled with CFSE. Cells were then transferred by the intravenous (i.v.) route to BALB/c recipients (n = 6) which were previously challenged (4 days) with DENV by the intracerebral (i.c.) route. Recipient mice also received 1 ml of anti-NS1 antiserum. After 3 days, recipients were sacrificed and spleen, blood and liver samples were analyzed by flow cytometry. To control the experiment, the same procedure was performed in parallel with splenocytes collected from non-vaccinated mice (n = 13).