Clonal Expansion of Infiltrating T Cells in the Spinal Cords of SJL/J Mice Infected with Theiler’s Virus

Abstract

Intracerebral infection of susceptible mice with Theiler’s murine encephalomyelitis virus results in immune-mediated inflammatory demyelination in the white matter and consequent clinical symptoms. This system has been utilized as an important virus model for human multiple sclerosis. Although the potential involvement of virus-specific Th cells has been studied extensively, very little is known about the nature of T cells infiltrating the CNS during viral infection and their role in the development of demyelinating disease. In this study, the clonal nature of T cells in the spinal cord during the disease course was analyzed using size spectratyping and sequencing of the TCR β-chain CDR3 region. These studies clearly indicate that T cells are clonally expanded in the CNS after viral infection, although the overall TCR repertoire appears to be diverse. The clonal expansion appears to be Ag-driven in that it includes Th cells specific for known viral epitopes. Interestingly, such restricted accumulation of T cells was not detectable in the infiltrates of mice with proteolipid protein peptide-induced experimental autoimmune encephalomyelitis. The initial T cell repertoire (7–9 days postinfection) seems to be more diverse than that observed in the later stage (65 days) of virally induced demyelination, despite the more restricted utilization of Vβ subfamilies. These results strongly suggest continuous stimulation and clonal expansion of virus-specific T cells in the CNS of Theiler’s murine encephalomyelitis virus-infected mice during the entire course of demyelinating disease.

Tim-3+ T-bet+ Tumor-Specific Th1 Cells Colocalize with and Inhibit Development and Growth of Murine Neoplasms

“…Growth of B lymphoma cells in mice injected with cells from a SJL/J Th1 clone specific for Theiler's virus peptide (9) was no different from that measured in control mice challenged with tumor cells alone. In this experiment the lymph nodes and spleens of tumor-injected control mice (n ϭ 6) were 3.57 and 3.50% of total body weight, respectively, and the lymph nodes and spleens of Ag-nonspecific, Th1-injected, tumor-challenged mice (n ϭ 8) were 4.71 and 4.25% of total body weight, respectively.…”

“…The relative concentrations of cDNA were estimated based on the level of ␤-actin amplification (35 cycles) by PCR. Each V␤ was assessed by PCR analysis using a sense sequence primer of the individual V␤s (V␤1, -2, -3, -4, -5, -6, -7, -10, -14, -15, -16, -17, -18, and -19) and a common antisense primer of downstream C␤ as previously described (9). The TCR V␤ chain oligonucleotide primer sequences are shown in Table II.…”

TCR Vβ8+ T Cells Prevent Development of Toxoplasmic Encephalitis in BALB/c Mice Genetically Resistant to the Disease

“…In relation to our finding of the importance of V␤8 ϩ T cells for genetic resistance of BALB/c mice against development of TE, an occurrence of clonal expansion of CD8 ϩ T cells was recently demonstrated in the brain of multiple sclerosis patients, by analyzing TCR V region sequences of T cells infiltrated into active lesions (31). In Theiler's murine encephalomyelitis virus-infected mice, a model of multiple sclerosis, clonal expansion of T cells was also suggested in demyelinating lesions in the brain (32). In a murine model of cerebral malaria, T cells bearing TCR V␤8.1 chain play a pathogenic role (33).…”

Predominant Clonal Accumulation of CD8+T Cells with Moderate Avidity in the Central Nervous Systems of Theiler's Virus-Infected C57BL/6 Mice

“…CDR3 length spectratyping was performed as previously described, with minor modifications (25,44). Primary amplification of cDNAs from isolated antigen-specific T cells was performed for 35 cycles using V␤-C␤ primer pairs at 58°C for annealing in the presence of PCR enhancer solution (GIBCO, Invitrogen Corporation, Grand Island, NY).…”

“…The PCR products were cloned into the pGEM-T easy vector (Promega, Madison, WI) and sequenced (Macrogen Co., Seoul, Korea, or Northwestern University genomic core facility). CDR3 size was defined as the number of residues between the aligned C (CASS) in the V␤ gene element and the FGXG in the J␤ region minus four (25).…”