It is suspected that SP plasma has strongly neutralizing antibodies (NAb) targeting specific HIV viral epitopes.
Methods:
NAbs levels of 11 HIV-1-infected SPs were detected by PBMC-based neutralization assays. To investigate SP NAb epitope, this study used a biopanning approach to obtain mimotopes of LEE011 clinical trial HIV-1 that were recognized by SP plasma NAbs. IgG was purified from high-titer NAb SP plasma, and used as the ligand for three rounds of biopanning to select HIV-specific mimotopes from a phage-displayed random peptide library. Double-antibody sandwich ELISA, competitive inhibition assays, and peptide sequence analysis were used to evaluate the characteristics of phage-borne mimotopes.
Results: SPs had significantly more plasma neutralizing activity than typical progressors (TPs) (p = 0.04). P2 and P9 plasma, which have highest-titer HIV-NAb, were selected as ligands for biopanning. After three rounds of biopanning, 48 phage clones were obtained, of which 22 clones were consistent with requirement, binding with HIV-1 positive plasma and unbinding with HIV-1 negative plasma. Compared with linear HIV-1 protein sequence and HIV-1 protein structure files, only
12 clones were selleck chemicals possible linear mimotopes of NAbs. In addition, the C40 clone located in gp41 CHR was found to be a neutralizing epitope, which could inhibit pooled HIV-1 positive plasma reaction.
Conclusion: Biopanning of serum IgG can yield mimotopes of HIV-1-related antigen SBI-0206965 in vivo epitopes. This methodology provides a
basis for exploration into HIV-1-related antigen-antibody interactions and furthers NAb immunotherapy and vaccine design. (C) 2012 Elsevier Editora Ltda. All rights reserved.”
“Background: Hypoplastic left heart syndrome (HLHS) is characterized by underdevelopment of the left ventricle (LV) and increased biomechanical stress on the right ventricle (RV) from single ventricle physiology Despite the clinical significance. the signaling pathways active during RV remodeling and disease progression are not known. To address this, we examined differential changes in expression of genes associated with transforming growth factor-beta (TGF-beta)/bone morphogenetic protein (BMP) signaling in RV tissue isolated from HLHS patients relative to RV and LV tissue from control subjects
Methods and Results: Quantitative real-time polymerase chain reaction was used to detect changes in expression of 84 genes involved in TGF-beta/BMP-mediated cardiac development, cell growth. and differentiation in RV tissue collected from 6 neonates with HLHS undergoing stage 1 Norwood procedure (age. 1-7 days. mean, 4 days) and RV and LV tissue obtained from 5 infants with noncardiac pathology (age range. 1-135 days mean.