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. 2021 May 18;12:2935. doi: 10.1038/s41467-021-23244-3

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© The Author(s) 2021, corrected publication 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a 5 mg MP containing 250 µg OVA and 2.5 µg polyI:C (MP-OVA/pIC) or 2.5 µg Riboxxim (MP-OVA/Rib) were incubated in PBS (pH 7.4) at 37 °C for a total period of 6 days. Amounts of encapsulated OVA and of OVA released from PLGA-MP at indicated time points were determined by SDS-PAGE and subsequent silver staining. Data are presented as means ± SD from three independent experiments. b Encapsulation efficiency and cumulative in vitro OVA release from 5 mg PLGA-MP-OVA/polyI:C (OVA/pIC, blue circles, n = 6) or PLGA-MP-OVA/Riboxxim (OVA/Rib, red squares, n = 6) over 6 days as analyzed by MicroBCA™ assay. c Near-infrared (NIR) fluorescence IVIS® images of a representative BALB/c mouse s.c. injected with 5 mg fluorescent MP-QD705/OVA/Rib at the indicated days post-immunization. Scaling of the pseudo-color code is depicted next to the luminescent images. d Ex vivo NIR fluorescence IVIS® image of draining popliteal and inguinal LN (dLNs) and of the contralateral non-draining LN (ndLN) of a representative mouse 6 days after s.c. immunization with MP-QD705/OVA/Rib. e Percentage of fluorescent particle internalization 6 h after incubating DCs with 10 µg/ml MP-QD705/OVA/Rib at 37 °C (black circles) or 4 °C (no uptake control, gray squares) was assessed by flow cytometry. Approximately 60% of PLGA-MP were efficiently engulfed by mouse bone marrow-derived DCs (mBMDCs, n = 3) or thioglycolate elicited peritoneal macrophages (mpMØ, n = 3), as well as by in vitro differentiated human monocyte-derived DCs (hMoDCs, n = 3) or monocyte-derived macrophages (hMoMØ, n = 3). PLGA particle uptake by human myeloid DCs was assessed using flow cytometry by gating on QD705⁺CD1c⁺ (n = 4) and QD705⁺CD141⁺ cells (n = 3) (black circles) after incubation of fluorescent microparticles with 10⁷ CD14-negative PBMCs. CD1c-negative (n = 4) or CD141-negative cells (n = 3) served as control (CTRL, gray squares). Data are presented as means ± SD and are derived from at least two independent experiments with a similar outcome.