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. Author manuscript; available in PMC: 2017 Jul 11.
Published in final edited form as: Sci Transl Med. 2016 May 25;8(340):340ra72. doi: 10.1126/scitranslmed.aaf1059

Fig. 3. Aβ's protective actions in cell culture are mediated by adhesion inhibition and agglutination activities against Candida.

Fig. 3

C. albicans adhesion to abiotic surfaces and agglutination in the bulk phase were characterized in the presence of cell-derived or synthetic Aβ. After 36 hours conditioning, host cell free culture media was collected from control non-transformed (H4-N or CHO-N) or transformed Aβ-overexpressing (H4-Aβ40, H4-Aβ42, or CHO-CAB) cultured cells. Aβ-immunodepleted (ID β-Aβ) and control immunodepleted ([ID IgG (immunoglobulin)] media were prepared by incubation with immobilized anti-Aβ or nonspecific antibodies. Experimental synthetic peptides included Aβ (Aβ40 and Aβ42), AMP positive control (LL-37), and negative control scrambled Aβ42 (scAβ42). (A and B) Comparison of ID β-Aβ and ID IgG media's adhesion inhibition (*P = 0.009, **P = 0.001, and ***P = 0.004) and agglutination (*P = 0.001, **P = 0.0005, and ***P = 0.004) activities. (C and D) Comparison of anti-Candida activities of serially diluted conditioned media and synthetic peptides. (E and F) Activities of synthetic Aβ42 monomer, soluble oligomeric amyloid-β derived diffusible ligands (ADDLs), and protofibril preparations. (G and H) Conditioned culture media adhesion inhibition (*P = 0.003 and **P < 0.0003) and agglutinating (*P < 0.02 and **P < 0.003) source activities alone (Neat) or in the presence of soluble yeast wall carbohydrates (+Glucan or +Mannan). (I) Synthetic monomeric Aβ42 and cell-generated peptide from H4-Aβ42 cells were compared for Candida binding using an Aβ/Candida binding ELISA. (J) Untreated, immunodepleted, or glucan (Glu)- or mannan (Man)-spiked H4-Aβ42 conditioned media were incubated with intact immobilized yeast cells in an Aβ/Candida binding ELISA assay (*P = 0.006, **P = 0.008, and ***P < 0.004). Synthetic peptide incubations (C to F and I) were performed in H4-Aβ42 conditioned culture media pre-treated to remove cell-derived Aβ by α-Aβ immunodepletion. Symbols and bars for (A) to (J) are statistical means of 6 replicate wells ± SEM. Statistical significance was by t-test.