A Novel Aza-MBP Altered Peptide Ligand for the Treatment of Experimental Autoimmune Encephalomyelitis

Abstract

Myelin basic protein (MBP) is a major target of T cells in lesions of multiple sclerosis (MS) patients and its animal model, experimental autoimmune encephalomyelitis (EAE). Interactions between the major histocompatibility complex II containing antigenic peptides and the T cell receptor activate CD4+ T cells that perpetuate EAE and MS. Previously reported data has shown that treating with an altered peptide ligand (APL) in which the normal antigenic peptide sequence of MBP has been slightly changed at T cell contact positions is helpful in reducing disease in both rodents and humans. The use of natural peptides, which are susceptible to protease degradation, requires high concentrations that can create hypersensitivity reactions. Our hypothesis is that APL containing aza substitutions, CH(R)-N- > N(R)N, could lead to improved protease resistance, reduced clinical disease scores, and a shift in T cell profile. In this study, several aza-APLs and control peptides were synthesized and screened for the best aza-APL candidate (3aza-APL) based on dissociation half time from major histocompatibility complex (MHC) class II, induction of IL-2 response, and resistance to degradation by proteases. The efficacy was then tested in vivo. Results indicate that 3aza-APL is superior to currently available APLs in terms of protease resistance and disease suppression in EAE mice. The 3aza-APL induced anti-inflammatory immune responses by altering key transcription factors and cytokine genes which regulate T cell subpopulations. These data suggest that the novel 3aza-APL has increased protease resistance property and is effective in reducing clinical and physiological signs of disease in EAE animals.

The Use of Electrochemical Voltammetric Techniques and High-Pressure Liquid Chromatography to Evaluate Conjugation Efficiency of Multiple Sclerosis Peptide-Carrier Conjugates

“…MS is regarded an autoimmune disease where immune cells (such as, Th1, Th17, macrophages, B cells) and their constituents (pro-inflammatory cytokines) are involved in the pathophysiology of the disease, with destruction of myelin sheath and loss of neurological function [ 5 , 6 , 7 , 8 , 9 , 10 ]. In an attempt to develop immunotherapeutics against MS using immunogenic/agonist peptides is to either alter the peptide to make it an antagonist [ 11 , 12 , 13 , 14 , 15 , 16 ], make it cyclic [ 17 , 18 , 19 ], or conjugate it to an appropriate carrier, which would deliver the peptide in such a manner to either induce tolerance, or alter the profile of T cells from pro-inflammatory (Th1) to anti-inflammatory (Th2) [ 13 , 14 , 15 ]. One approach which our team has developed, is to use mannan, a poly-mannose carrier conjugated to MS peptides [ 20 , 21 , 22 , 23 , 24 ].…”

Differential CpG DNA methylation of peripheral B cells, CD4+ T cells, and salivary gland tissues in IgG4-related disease

“…MBP (cg18455083) was hypomethylated in CD4 + T cells and salivary gland tissues of IgG4-RD patients. MBP encodes for myelin basic protein, which is the major target of T cells in lesions of multiple sclerosis (MS) patients and the animal model of experimental autoimmune encephalomyelitis [ 31 ]. Numerous studies have demonstrated the role of myelin reactivity of T cells in MS [ 32 – 34 ].…”

Differential CpG DNA Methylation of Peripheral B, CD4 + T cells and Salivary Gland Tissues in IgG4-related Disease

“…We found that MBP (cg18455083) was uniformly hypomethylated in B cells, CD4 + T cells and tissue of IgG4-RD patients compared with controls. Myelin basic protein (MBP) is the major target of T cells in lesions of multiple sclerosis (MS) patients and its animal model, experimental autommune encephalomyelitis (EAE) 23 . A number of previous studies have demonstrated the role of myelin-reactivity of T cells in MS 24,25,26 .…”