|PDF Version||Antiviral Cell Immunology|
|Director : François Lemonnier (email@example.com)|
The work of the laboratory aims, using different approaches, especially the production of genetically modified strains of mice (knock-out mice lacking selectively or globally H-2 class I or class II molecules, HLA class I or class II transgenic mice) at a) the analysis of the mechanisms involved in the education, the peripheral mobilization and the long term maintenance of cytolytic CD8+, helper CD4+ T lymphocytes and Natural Killer (NK) cells, b) the identification of viral or tumoral peptides presented to cytolytic and helper T lymphocytes, c) the evaluation and optimization of their vaccine potential and d) finally, the evaluation of the auto agressive potential.
Introduction of cytolytic T lymphocytes recognizing a histocompatibility class Ib molecule deprived of bound peptide : HFE (P. Rohrlich, H. Firat)
Immunizing mice expressing a single type of HLA class I molecule (HLA-A0201), with transfected cells expressing also a single species of HLA class Ib molecule (the human HFE molecule implicated in the control of iron metabolism), CD8+, TcRß+ cytolytic T lymphocytes recognizing specifically HFE through their T cell receptor for the antigen, were induced despite the fact that HFE molecules, as documented crystographically, are totally deprived of bound peptide. Similar CTL responses have been induced in DBA/2 HFE KO mice against the mouse HFE molecule. In view of the structural polymorphism in humans of the HFE molecule, these results support the possibility that HFE could be a significant histocompatibility antigen worth to consider in certain tissue transplantations.
Cytolytic T cell responses against leukemic cells (H. Firat, P. Langlade-Demoyen)
The junctional sequences corresponding to the 12-21 translocation (acute lymphoblastic leukemia) and the Philadelphia translocation (myeloblastic leukemia) give rise to neo antigenic peptides which can be presented to CD8+ cytolytic T cells by some HLA class I molecules and can induce in patients leukemia cell specific cytolytic responses.Using HLA-class I transgenic, H-2 class I KO mice, strategies susceptible to induce potent leukemic cell specific cytolytic responses were comparatively evaluated. The induction in patients of such responses might be of interest to reduce the risk of leukemic relapse.
Vaccine potential of HLA-A0201- and HLA-B702-restricted epitopic peptides of viral or human tumor origin (P. Langlade-Demoyen, S. Cardinaud, P. Rohrlich)
The vaccine potential of HIV 1, CMV and human tumor-derived epitopic peptides reported in the literature as efficiently presented by either HLA-A0201 or HLA-B0702 molecules have been compared using transgenic mice expressing HLA-A0201 molecules in the complete absence of H-2 class I molecules or the HLA-B0702 molecules in a H-2Kb, H-2Db double KO context. Following these studies, it was possible to establish a hierarchy in terms of immunogenicity of these peptides, to optimize the immunogenicity of those which were poorly immunogenic in their native configuration, to compare vaccine strategies and to demonstrate that a polyepitope construct could induce multiepitopic, long-lasting cytolytic responses in a single mouse. The studies currently performed have resulted in the identification of new epitopic peptides in the viral proteins (TAT, REV, VPU, VPR) expressed in the early phase of the HIV 1 replication cycle. These studies should result in the construction of a polyallelic (HLA-A02.01, HLA-B07.02) polyepitope construct of inter est for 60% of the individuals of the different human populations.
Creation of new HLA-class I transgenic, H-2 class I KO and HLA class II transgenic, H-2 class II KO mice (A. Pajot, Y.C. Lone)
Monochain constructs (by covalent linkage of human ß2-microglobulin to the first domain of HLA class I heavy chain) have been realized for the HLA-A*01.03, -A*03.01, -A*24.02, HLA-B*08.01, -B*27.05, -B*35.01, -B*44.02, HLA-Cw*07.01, -Cw*07.02 genes. The functionality of these constructs has been checked by transfection of cells and they are currently introduced in mouse oocytes to create the corresponding transgenic strains.
Constructions coding for the HLA class II DPß401, DP103 have been realized to derive HLA class II transgenic, H-2 class II KO mice in order to identify HIV 1-derived epitopic peptides presented by these DP molecules which are expressed by approximately 60% of individuals in human populations. Similarly, transgenic (HLA-A0201, HLA-DR) H-2 class I class II KO mice have been derived. These animals will be used to evaluate preclinically and simultaneously all components of the immune adaptative responses (CD8+ cytolytic, CD4+ helper and antibody) to vaccine formulations of potential interest in humans.
Genes controlling NK cell differentiation (C. Roth)
Using stable NK cell lines and clones derived from P53 deficient mice, two complementary approaches have been used to identify new receptors and co-receptors potentially involved in NK cell maturation. A differential expression library was prepared from clones either highly or poorly cytotoxic. Among the different cDNA isolated, we are focusing on those expressed by highly cytolytic NK cells. In collaboration with the proteomic group (PT3-genopole), we are identifying biochemically the peptides and proteins over-expressed in the highly cytotoxic clones. The biological validation of the candidate molecules identified is currently under investigation using genetically modified strains of mice.
Keywords: HLA Transgenic, H-2 KO mice, cancer, HIV 1, NK
|Publications of the unit on Pasteur's references database|
|Office staff||Researchers||Scientific trainees||Other personnel|
|LEMONNIER François, DR1 INSERM, Chef d’Unité IP,firstname.lastname@example.org
LANGLADE-DEMOYEN Pierre, Chef de Laboratoire IP,email@example.com
LONE Yu Chun , Chargé de Recherche CNRS,firstname.lastname@example.org
CHENIER Rémi, CR IP,email@example.com
ROTH Claude, CR IP,firstname.lastname@example.org
|CARDINAUD Sylvain, student
MATTEI Stefano, post-doctoral fellow
PAJOT Anthony, thesis student
ROHRLICH Pierre, Docteur en Médecine, AP-HP
|GARCIA-PONS, Francisco, Research Engineer IP|