Expression, purification, crystallization and preliminary X-ray diffraction analysis of swine leukocyte antigen 2 complexed with a CTL epitope AS64 derived from Asia1 serotype of foot-and-mouth disease virus

Abstract

BackgroundCurrently, the structural characteristics of the swine major histocompatibility complex (MHC) class I molecule, also named swine leukocyte antigen class I (SLA-I) molecule need to be further clarified.ResultsA complex of SLA-I constituted by an SLA-2*HB01 molecule with swine β2-microglobulin and a cytotoxic T lymphocyte (CTL) epitope FMDV-AS64 (ALLRSATYY) derived from VP1 protein (residues 64–72) of Asia 1 serotype of foot-and-mouth disease virus (FMDV) was expressed, refolded, purified and crystallized. By preliminary X-ray diffraction analysis, it was shown that the diffraction resolution of the crystal was 2.4 Å and the space group belonged to P212121 with unit cell parameters a = 48.37, b = 97.75, c = 166.163 Å.ConclusionThis research will be in favor of illuminating the structural characteristics of an SLA-2 molecule associated with a CTL epitope derived from Asia1 serotype of FMDV.

Advanced Foot-And-Mouth Disease Vaccine Platform for Stimulation of Simultaneous Cellular and Humoral Immune Responses

“…Currently, FMD vaccine studies using an inactivated vaccine, modified virus inactivated vaccine (DIVA vaccine) [ 24 ], viral vector vaccine [ 25 ], adenovirus [ 26 , 27 , 28 ], virus-like particle (VLP) vaccine (bacterial) [ 29 ], baculovirus [ 30 , 31 ], plant-produced empty capsids [ 32 , 33 ], peptide vaccine (Poly(I:C)) [ 34 ], CpG [ 35 ], plant based recombinant vaccine (alfalfa [ 36 ], chloroplast of tobacco [ 37 ]), RNA vaccine [ 38 ], DNA vaccine (cDNA [ 39 , 40 ], electroporation [ 41 ], T cell [ 42 , 43 , 44 ], B cell [ 45 , 46 ] epitope, APC targeting [ 47 ]), live attenuated vaccine [ 48 , 49 , 50 , 51 ], and antiviral agent [ 52 , 53 ] are underway. However, no high-quality FMD vaccines that surpass the efficacy of the inactivated vaccine have been successfully developed to date.…”

The Parallel Presentation of Two Functional CTL Epitopes Derived from the O and Asia 1 Serotypes of Foot-and-Mouth Disease Virus and Swine SLA-2*HB01: Implications for Universal Vaccine Development

“…The preparation, refolding, and crystallization of SLA-2*HB01 and sβ 2 m proteins complexed with peptides were performed as previously reported [ 35 , 39 ]. In this research, additional peptides (shown in Table 1 ) were used in refolding and crystallization with SLA-2*HB01 and sβ 2 m proteins.…”

“…In this research, additional peptides (shown in Table 1 ) were used in refolding and crystallization with SLA-2*HB01 and sβ 2 m proteins. Two peptides, Hu64, which is derived from the O serotype of FMDV Tibet/CHA/99 (HuBHK99), and As64, from 1/Jiangsu/China/2005, were crystallized with SLA-2*HB01 and sβ 2 m, and their crystal data were collected as previously reported [ 35 , 39 ].…”

“…It was shown that the PBG of SLA-3 had different peptide-binding preferences to those of SLA-1, especially in the P2, P3, and P9 positions. Up to now, the three-dimensional structures of SLA-2 molecules have been identified, and the crystal of SLA-2*HB01 has been described, albeit without structural information [ 33 , 34 , 35 ]. Specifically, there has been insufficient investigation into the structural characteristics of SLA-2 molecules.…”

Peptide-Based Vaccines: Foot-and-Mouth Disease Virus, a Paradigm in Animal Health

“…For this reason, CTL epitope characterization has been mostly addressed by in silico predictions and inbred mice, with the understanding that such models need to be confirmed experimentally. For example, Barfoed et al reported a murine H2-Kd-restricted CTL epitope that induced cell immunity in mice but did not achieve a protective effect in other animals [ 104 ]; in contrast, Gao et al described several 9-residue peptides from FMDV VP1 that not only induced CTL response in mice but also provided protective response in guinea pigs [ 105 , 106 , 107 ].…”