Journal: iScience
Article Title: In vitro recapitulation of intramuscular mRNA vaccination with naive and recall antigens using a human lymphoid follicle chip platform
doi: 10.1016/j.isci.2026.116416
Figure Lengend Snippet: Induction of LFs and anti-RABV-G IgG production by SAM-LNP1 in human LF chips (A) Schematic of the LF Chip created with monocytes and conditioned medium from the intramuscular vaccination-mimicking module created with BioRender.com . (B) Representative 3D confocal microscopic stacks showing pseudo-colored follicles (blue) and cell nuclei (green) present within extracellular matrix (ECM) gels cultured for 4 days within a perfused LF Chip when vaccinated with SAM-LNP1 in the absence or presence of IL-2 and IL-4 (SAM-LNP1 + IL-2&IL-4); scale bars, 100 μm. (C) Quantification of the number (left) and size (right) of LFs in LF chips of one donor based on immunostaining followed by confocal imaging. Each data point represents one field of view (left) or an individual follicle (right); 2 independent chips were analyzed per condition. (D) Anti-RABV-G IgG levels in effluents of no treatment LF chips or chips vaccinated with or without IL-2 and IL-4 and cultured for 14 days were detected using a cell-based assay; 3–4 independent chips were analyzed per condition. (E) Quantification of the number (left) and size (right) of LFs in LF chips from four different donors, with each color representing a different donor. Each data point corresponds to one field of view (left) or an individual follicle (right). (F) Anti-RABV-G IgG levels in effluents of LF chips. Each data point represents one chip, with different symbols indicating chips from three independent donors. (G) Anti-RABV-G IgG levels in effluents of LF chips seeded with naive B + bulk T + monocyte mixture. Each data point represents one chip, with different symbols indicating chips from three independent donors. (H) Heatmap showing average log2 fold changes in cytokine levels in the effluents of three LF chips created using bulk lymphocytes from donors whose prior exposure to rabies was unknown, measured using a Luminex Multiplex Assay at various time points (4, 7, 15, 22, and 28 days post-vaccination) compared to the average levels of each cytokine found at day 1. Representative results from one donor are shown in (B)–(D) and (H), with similar outcomes observed in two donors. Data shown are mean ± SD; (C, left; D; and E) one way ANOVA test followed by Tukey’s multiple comparisons test, (C, right; and E) Mann-Whitney U test, (F and G) Welch’s t test.
Article Snippet: After allowing the matrix to gel for 30 min at 37°C, the top channel was filled with 1 part conditioned medium from the 2D intramuscular module, and 2 parts fresh medium supplemented with IL-2 (Mitenyi Biotec, 130-097-742) and IL-4 (Mitenyi Biotec, 130-093-922) for experiments with RABV-G mRNA vaccines.
Techniques: Cell Culture, Immunostaining, Imaging, Cell Based Assay, Luminex, Multiplex Assay, MANN-WHITNEY
Miltenyi Biotec is a verified supplier 
Miltenyi Biotec manufactures this product 
