I-Domain-Antigen Conjugate (IDAC) for Delivering Antigenic Peptides to APC: Synthesis, Characterization, and in Vivo EAE Suppression

详细信息    查看全文

• 作者：Prakash Manikwar ; Barlas B眉y眉ktimkin ; Paul Kiptoo ; Ahmed H. Badawi ; Nadezhda A. Galeva ; Todd D. Williams ; Teruna J. Siahaan
• 刊名：Bioconjugate Chemistry
• 出版年：2012
• 出版时间：March 21, 2012
• 年：2012
• 卷：23
• 期：3
• 页码：509-517
• 全文大小：458K
• 年卷期：v.23,no.3(March 21, 2012)
• ISSN：1520-4812

文摘

The objectives of this work are to characterize the identity of I-domain鈥揳ntigen conjugate (IDAC) and to evaluate the in vivo efficacy of IDAC in suppressing experimental autoimmune encephalomyelitis (EAE) in mouse model. The hypothesis is that the I-domain delivers PLP_139鈥?51 peptides to antigen-presenting cells (APC) and alters the immune system by simultaneously binding to ICAM-1 and MHC-II, blocking immunological synapse formation. IDAC was synthesized by derivatizing the lysine residues with maleimide groups followed by conjugation with PLP-Cys-OH peptide. Conjugation with PLP peptide does not alter the secondary structure of the protein as determined by CD. IDAC suppresses the progression of EAE, while I-domain and GMB-I-domain could only delay the onset of EAE. As a positive control, Ac-PLP-BPI-NH₂-2 can effectively suppress the progress of EAE. The number of conjugation sites and the sites of conjugations in IDAC were determined using tryptic digest followed by LC-MS analysis. In conclusion, conjugation of I-domain with an antigenic peptide (PLP) resulted in an active molecule to suppress EAE in vivo.