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. 2021 Nov 10;13(11):1904. doi: 10.3390/pharmaceutics13111904

Figure 7.

Figure 7

Antibody distribution is dependent on the presence of the antigen in the CNS and the presence of an Fc domain (study 3). The distribution and clearance in the murine OM and CNS are shown 45 min and 20 h (E,J,O,T) after directed intranasal administration at the olfactory region (OM). All images were captured with a confocal microscope. Arrowheads point to distinct stained structures while asterisk show diffuse staining pattern. (A–E) Background signal due to endogenous IgGs, that have not been cleared by transcardial perfusion, and autofluorescence in the tissue of an animal, which received a vehicle control (PBS). (F–J) distribution of anti-Nogo-A monoclonal full antibody 11C7. The 11C7 was detected in the OM (F), the OB (G), the AON/OT (anterior olfactory nucleus/olfactory tubercle) (H), the choroid plexus in the ventricles (I), and after 20 h in the hippocampus (J). The strongest signal was detected in the olfactory epithelium followed by the olfactory bulb and the choroid plexus. In the hippocampus, the signals were amplified for a better visualization. (K–O) Distribution of the isotype control (IC) antibody, which does not recognize any structure of the murine CNS as antigen. The distribution profile within 45 min was similar to 11C7 however the immunoreactivity in the AON/OT was lower in all animals investigated. The similar distribution pattern in OM, OB, AON and the subventricular zones imply the relevance of the Fc receptor system. The absence of IC in hippocampus (O) is a strong indication that the presence of the antigen is critical to avoid rapid elimination. (P–T) A scFv format of 11C7 was distributed to the OB to a lesser extent than the IgGs. In addition, hardly any signals higher than the vehicle control could be found in the AON/OT, choroid plexus, subventricular zones, nor in the Nogo-A expressing cells in the hippocampus. Nevertheless, since the 11C7 scFv was detected via its penta His-Tag, the intensities of (A–O) should not be directly compared with (P–T). Representative images are shown. OM, olfactory mucosa; OB, olfactory bulb; AON, anterior olfactory nucleus; OT, olfactory tubercle. Scale bar: 100 µm.