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. Author manuscript; available in PMC: 2016 Jan 31.
Published in final edited form as: J Immunol. 2014 Dec 29;194(3):1047–1056. doi: 10.4049/jimmunol.1401832

Figure 5. CD40L-induced reticulation facilitates the intercellular transfer of exogenous Ag between DC1.

Figure 5

(A–C) Cy5-labeled (blue), YG latex bead (40 nm)-containing ‘donor’ DC1 or DC2 were co-cultured with the respective non-bead-containing, Cy3-labeled (red) ‘recipient’ DC type for 20 h in the presence or absence of rhCD40L. Data are representative of 2 donors independently tested. (A) Montage of time-lapse, confocal imaging (600×) showing beads (green; arrows) moving rapidly through donor DC1 TNTs (blue) over a time span of 28 min. See also Video 5. (B) Montage tracing the pathway of beads (green; arrows) traveling from the cell body of donor DC1 (blue), through donor cell TNTs and into the connected recipient DC (red), where they finally collect in the recipient cell body (yellow, lower left arrows). (C) Flow cytometric analysis and quantification of intercellular exogenous bead transfer from donor to recipient DC types after treatment with media only or CD40L. (D) IFNγ-ELISPOT assays measuring Ag-specific recall responses of cultured T cells following their in vitro sensitization with FACS sorted, YG bead/Ag positive and negative recipient DC1. Data are represented as mean ± SE of 2 independent experiments, and * represents the p-value <0.05. See also Supplemental Fig. 3.