Fig. 3: ATVTfR and ATVCD98hc exhibit enhanced brain uptake and biodistribution compared to control IgG. | Nature Communications

Fig. 3: ATVTfR and ATVCD98hc exhibit enhanced brain uptake and biodistribution compared to control IgG.

From: Fc-engineered large molecules targeting blood-brain barrier transferrin receptor and CD98hc have distinct central nervous system and peripheral biodistribution

Fig. 3

a Schematic of the experimental design. WT, TfRmu/hu KI, and CD98hcmu/hu KI mice were dosed with 50 mg/kg of AF647-conjugated control IgG and ATVTfR for 1 day and ATVCD98hc for 5 days. b Brain concentration of AF647-conjugated control IgG, ATVTfR, or ATVCD98hc as measured by huIgG ELISA in bulk brain lysates after a single 45 mg/kg IV dose. n = 4/ group, mean ± SEM, one-way ANOVA ****p < 0.0001 compared to control IgG, 95% confidence interval of ATVTfR vs. control IgG −34.76 to −23.5, 95% confidence interval of ATVCD98hc vs. control IgG −24.72 to −13.47. c Representative immunofluorescence in sagittal brain sections of AF647-conjugated control IgG, ATVTfR, and ATVCD98hc in mouse. Representative immunostainings from n = 2/group. d Brain lysate concentration measured by ELISA 1 day after a single 100 mg/kg IV dose of control IgG or 15 mg/kg of ATVTfR (n = 4 mice/group), unpaired two-tailed t-test, nonsignificant (n.s.) p value > 0.05. e Immunodetection of huIgG in whole sagittal brain sections by widefield imaging. Experiment conducted once with n = 4/group. f Magnified examples of brain regions from (e) including the superior colliculus, neocortex, and thalamus. Examples of putative perivascular huIgG signal indicated by arrowheads; notably darker alpha-smooth muscle actin (αSMA) + arteriole indicated by arrows (see Supplementary Fig. 4e–g). Insets show immunodetected huIgG (magenta) and vascular marker caveolin-1 (green), scalebar 50 μm. g Ventral volume view of immunodetected huIgG in tissue-cleared whole mouse hemibrain (n = 6 mice/group). Arrowheads indicate huIgG associated with the large surface arteries, asterisk indicates the median eminence, arrow indicates cranial nerves (region of trigeminal, facial, and vestibulocochlear nerves). h Higher magnification light sheet imaging of immunodetected huIgG and vascular marker lectin around the middle cerebral artery (MCA) and anterior cerebral artery (ACA) branching from the circle of Willis. Circumferential banding pattern around putative smooth muscle cells indicated by arrowheads; arrow indicates putative perivascular profile of a penetrating vessel. For micrographs, display settings were optimized independently for each region of interest and were identical for both treatment groups.

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