Long-term human coronavirus-myelin cross-reactive T-cell clones derived from multiple sclerosis patients

Abstract

Autoimmune reactions associated with MS involve genetic and environmental factors. Because murine coronaviruses induce an MS-like disease, the human coronaviruses (HCoV) are attractive candidates as environmental factors involved in a demyelinating pathology. We previously reported the isolation of HCoV-229E/myelin basic protein (MBP) cross-reactive T-cell lines (TCL) in MS patients. To investigate antigenic cross-reactivity at the molecular level, 155 long-term T-cell clones (TCC) were derived from 32 MS patients by in vitro selection with MBP, proteolipid protein (PLP) or HCoV (strains 229E and OC43). Overall, 114 TCC were virus-specific, 31 were specific for myelin Ag and 10 other were HCoV/myelin cross-reactive. Twenty-eight virus-specific TCC and 7 myelin-specific TCC were obtained from six healthy donors. RACE RT-PCR amplification of the Vbeta chains of five of ten the cross-reactive TCC confirmed clonality and sequencing identified the CDR3 region associated with cross-reactivity. Our findings have promising implications in the investigation of the role of molecular mimicry between coronaviruses and myelin in MS as a mechanism related to disease initiation or relapses.

Rapid detection of clonal expansion of T‐cell receptor‐beta gene in patients with HBV using the real‐time PCR with DNA melting curve analysis

“…Many methods have been used to detect the composition of the TCRBV CDR3 gene in PBMCs, such as Southern blots, denaturing gradient gel electrophoresis (DGGE) and the use of monoclonal antibodies 20 . Recently, fluorescent PCR with gene scanning technology has been extensively used to analyze the expression frequency and the length of TCRBV CDR3 gene, thereby reflecting the status of αβ+ T cells clonal proliferation 21,22 . Subsequently, real time PCR with melting curve analysis in the LightCycler system (Roche, Indianapolis, IN) has been used for in vitro analysis of cloned immunoglobulin (Ig) or TCR gene rearrangements using genomic DNA as PCR template 23,24 .…”

The preparation and comparison of decellularized nerve scaffold of tissue engineering

“…Next, goat anti-rabbit IgG antibody-horseradish peroxidase polymer at 37 C for 2 h, stained by diaminobenzidine, and sealed by neutral gum.…”

Regulatory T Cells Selectively Preserve Immune Privilege of Self-Antigens during Viral Central Nervous System Infection

“…A number of viruses, including EBV (3), human herpes virus-6 (4), human coronavirus OC-43 (5), and human coronavirus 229E (6), have been found in the CNS of multiple sclerosis patients. Furthermore, durable immune responses against these viruses have been detected in affected individuals (4,7,8). Infection with neurotropic viruses may not only induce specific antiviral immune responses in the CNS, but also most likely predisposes to a stereotyped immunopathological tissue response that precipitates de novo immune reactions against genuine self-antigens (2,9).…”