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Rôle des cellules T CD8+ dans la pathogenèse de l'hépatite auto-immune : développement d'un modèle murin transgénique
| Content Provider | Semantic Scholar |
|---|---|
| Author | Fakhfakh, Amin |
| Copyright Year | 2007 |
| Abstract | Backgrottnd: Autoimmune hepatitis (AIH) is a disease of unknown etiology characterized by the progressive destruction of the hepatic parenchyrna, following lymphocyte infiltration. Thus far, no animal mode! developing a spontaneous and chronic AIH as observed in hurnans bas been characterized. Ami: Our aim is to produce a double transgenic (1g) murine mode! expressing the LCMV’s nucleoprotein (NP) and a TCR specific to its immunodorninant epitope, in order to induce a spontaneous AIH. Methods: The phenotype of clone NPI8 (CD8+ specific against NP peptide 396-404) TCR’s Œ and 3 chains was determined by RT-PCR using specific primers. PCR products ofthese two 1CR chains were cloned into two specific vectors (cassettes) allowing their expression in transgenic mice. Expression of the transgenes by these cassettes was assessed by immunofluorescence, by specific RT-PCR and by flow cytometry on transfected T celis defective for TCRŒ and TCRf3 chains (58 -I-). The functionality of this 1CR was checked for production ofthe IL-2 by ELISA. Resuits: TCRu and TCRJ3 chains ofcloneNPl8 were detemiined as being VŒ8.5-JŒ6.19 and Vf312-D132-Jf32.3 by sequencing of the PCR products and comparison with sequences in databanks. The expression ofthe 1CR by transfected 58 -7cells was tested by flow cytornetry, and then confirmed by immunofluorescence and RT-PCR. Our results show that sorne of the transfected 58-/produced IL-2 after stimulation with splenic celis charged with peptide NP396404. Conclusion: The two TCRŒ and TCR3 transgenes have been used to produce an anti NP396404 TCR Tg mouse that will be crossed with a 1g mouse expressing NP in the liver. The resultant double Tg mouse wiÏl allow the study ofmechanisrns undertying the deveÏoprnent of a spontaneous AIH through the destruction of hepatocytes by CD8+ T lymphocytes. |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://papyrus.bib.umontreal.ca/xmlui/bitstream/handle/1866/15170/Fakhfakh_Amin_2006_memoire.pdf?isAllowed=y&sequence=1 |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
