The JHM strain of mouse hepatitis virus induces a spike protein-specific Db-restricted cytotoxic T cell response

185

282

A chronic demyelinating disease results from murine infection with the neurotropic strain JHM of mouse hepatitis virus (MHV-JHM). Demyelination is largely immune mediated. In this study, the individual roles of CD4 and CD8 T cells in MHV-induced demyelination were investigated using recombination-activating gene 1−/− (RAG1−/−) mice infected with an attenuated strain of MHV-JHM. These animals develop demyelination only after adoptive transfer of splenocytes from mice previously immunized to MHV. In this study, we show that, following adoptive transfer, virus-specific CD4 and CD8 T cells rapidly infiltrate the CNS of MHV-JHM-infected RAG1−/− mice. Adoptive transfer of CD4 T cell-enriched donors resulted in more severe clinical disease accompanied by less demyelination than was detected in the recipients of undepleted cells. Macrophage infiltration into the gray matter of CD4 T cell-enriched recipients was greater than that observed in mice receiving undepleted splenocytes. In contrast, CD8 T cell-enriched recipients developed delayed disease with extensive demyelination of the spinal cord. MHV-JHM-infected RAG1−/− mice receiving donors depleted of both CD4 and CD8 T cells did not develop demyelination. These results demonstrate that the development of demyelination following MHV infection may be initiated by either CD4 or CD8 T cells. Furthermore, they show that CD4 T cells contribute more prominently than CD8 T cells to the severity of clinical disease, and that this correlates with increased macrophage infiltration into the gray matter.

Section: Normal T and B Cells Do Not Develop In Scid And Rag1mentioning

confidence: 99%

CD4 and CD8 T Cells Have Redundant But Not Identical Roles in Virus-Induced Demyelination

Wu¹,

Dandekar

Pewe

et al. 2000

185

282

“…Splenocytes were isolated at 8 days p.i. and pooled, and 1 ϫ 10 6 total cells were stimulated with peptide corresponding to the CD8 epitope in the surface (S) glycoprotein spanning residues 510 -518 (S510 -518) (48,49). After incubation for 6 h at 37°C in medium containing Golgi stop (Cytofix/Cytoperm kit; BD PharMingen), cells were washed, and blocked with PBS containing 10% FBS and a 1/200 dilution of CD16/32 (BD PharMingen).…”

Section: Intracellular Cytokine Stainingmentioning

confidence: 99%

IFN-γ-Inducible Protein 10 (IP-10; CXCL10)-Deficient Mice Reveal a Role for IP-10 in Effector T Cell Generation and Trafficking

Dufour

Dziejman

Liu

et al. 2002

971

713

IFN-γ-inducible protein 10 (IP-10, CXCL10), a chemokine secreted from cells stimulated with type I and II IFNs and LPS, is a chemoattractant for activated T cells. Expression of IP-10 is seen in many Th1-type inflammatory diseases, where it is thought to play an important role in recruiting activated T cells into sites of tissue inflammation. To determine the in vivo function of IP-10, we constructed an IP-10-deficient mouse (IP-10−/−) by targeted gene disruption. Immunological analysis revealed that IP-10−/− mice had impaired T cell responses. T cell proliferation to allogeneic and antigenic stimulation and IFN-γ secretion in response to antigenic challenge were impaired in IP-10−/− mice. In addition, IP-10−/− mice exhibited an impaired contact hypersensitivity response, characterized by decreased ear swelling and reduced inflammatory cell infiltrates. T cells recovered from draining lymph nodes also had a decreased proliferative response to Ag restimulation. Furthermore, IP-10−/− mice infected with a neurotropic mouse hepatitis virus had an impaired ability to control viral replication in the brain. This was associated with decreased recruitment of CD4+ and CD8+ lymphocytes into the brain, reduced levels of IFN-γ and the IFN-γ-induced chemokines monokine induced by IFN-γ (Mig, CXCL9) and IFN-inducible T cell α chemoattractant (I-TAC, CXCL11) in the brain, decreased numbers of virus-specific IFN-γ-secreting CD8+ cells in the spleen, and reduced levels of demyelination in the CNS. Taken together, our data suggest a role for IP-10 in both effector T cell generation and trafficking in vivo.

“…Intracellular staining for IFN-␥ was performed using a total of 10 6 cells stimulated separately for 6 h with either the CD4 or CD8 epitopes M133-147 and S510 -518, respectively, and stained for intracellular IFN-␥ using PE-conjugated anti-IFN-␥ (1:50; XMG1.2; BD PharMingen, San Diego, CA) for 1 h at 4°C (35)(36)(37)(38)(39)(40). Additional Abs used for immunophenotyping cells in these studies include allophycocyanin-conjugated rat anti-mouse CD4 (BD PharMingen), allophycocyanin-conjugated rat anti-mouse CD8 (BD PharMingen), FITC-conjugated rat anti-mouse F4/80 (Serotec, Oxford, U.K.), and PE-conjugated rat anti-mouse CD45 (BD PharMingen) (36,37).…”

Section: Mononuclear Cell Isolation and Intracellular Cytokine Stainingmentioning

confidence: 99%

Antibody Targeting of the CC Chemokine Ligand 5 Results in Diminished Leukocyte Infiltration into the Central Nervous System and Reduced Neurologic Disease in a Viral Model of Multiple Sclerosis

Glass

Hickey

Hardison

et al. 2004

120

111

Intracerebral infection of mice with mouse hepatitis virus, a member of the Coronaviridae family, reproducibly results in an acute encephalomyelitis that progresses to a chronic demyelinating disease. The ensuing neuropathology during the chronic stage of disease is primarily immune mediated and similar to that of the human demyelinating disease multiple sclerosis. Secretion of chemokines within the CNS signals the infiltration of leukocytes, which results in destruction of white matter and neurological impairment. The CC chemokine ligand (CCL)5 is localized in white matter tracts undergoing demyelination, suggesting that this chemokine participates in the pathogenesis of disease by attracting inflammatory cells into the CNS. In this study, we administer a mAb directed against CCL5 to mice with established mouse hepatitis virus-induced demyelination and impaired motor skills. Anti-CCL5 treatment decreased T cell accumulation within the CNS based, in part, on viral Ag specificity, indicating the ability to differentially target select populations of T cells. In addition, administration of anti-CCL5 improved neurological function and significantly (p ≤ 0.005) reduced the severity of demyelination and macrophage accumulation within the CNS. These results demonstrate that the severity of CNS disease can be reduced through the use of a neutralizing mAb directed against CCL5 in a viral model of demyelination.