A Lentiviral Vector Allowing Physiologically Regulated Membrane-anchored and Secreted Antibody Expression Depending on B-cell Maturation Status.
The development of lentiviral vectors (LVs) for expression of a specific antibody can be achieved through the transduction of mature B-cells. This approach would provide a versatile tool for active immunotherapy strategies for infectious diseases or cancer, as well as for protein engineering. Here, we created a lentiviral expression system mimicking the natural production of these two distinct immunoglobulin isoforms. We designed a LV (FAM2-LV) expressing an anti-HCV-E2 surface glycoprotein antibody (AR3A) as a membrane-anchored Ig form or a soluble Ig form, depending on the B-cell maturation status. FAM2-LV induced high-level and functional membrane expression of the transgenic antibody in a nonsecretory B-cell line. In contrast, a plasma cell (PC) line transduced with FAM2-LV preferentially produced the secreted transgenic antibody. Similar results were obtained with primary B-cells transduced ex vivo. Most importantly, FAM2-LV transduced primary B-cells efficiently differentiated into PCs, which secreted the neutralizing anti-HCV E2 antibody upon adoptive transfer into immunodeficient NSG (NOD/SCIDγc(-/-)) recipient mice. Altogether, these results demonstrate that the conditional FAM2-LV allows preferential expression of the membrane-anchored form of an antiviral neutralizing antibody in B-cells and permits secretion of a soluble antibody following B-cell maturation into PCs in vivo.
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France.
Floriane
F
Fusil
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France.
Sara
S
Calattini
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France.
Fouzia
F
Amirache
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France.
Jimmy
J
Mancip
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France.
Caroline
C
Costa
Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA; Present address: Organovo Holdings, Inc., San Diego, California, USA.
Justin B
JB
Robbins
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France; Present address: Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA.
Florian
F
Douam
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France; Present address: Institut Pasteur Shanghai, Chinese Academy of Sciences, Shanghai, China.
Dimitri
D
Lavillette
Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, USA.
Mansun
M
Law
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France.
Thierry
T
Defrance
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France; Inserm, U1065, Centre Méditerranéen de Médecine Moléculaire (C3M), équipe "contrôle métabolique des morts cellulaires", Nice, France.
Els
E
Verhoeyen
CIRI, International Center for Infectiology Research, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France; Université Claude Bernard Lyon 1, Centre International de Recherche en Infectiologie, Lyon, France; CNRS, UMR5308, Lyon, France; LabEx Ecofect, Université de Lyon, Lyon, France. Electronic address: flcosset@ens-lyon.fr.
François-Loïc
FL
Cosset
D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000920:Antibody-Dependent Cell Cytotoxicity; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D005822:Genetic Vectors; D057809:HEK293 Cells; D016174:Hepacivirus / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D016086:Lentivirus; D008213:Lymphocyte Activation; D008562:Membrane Glycoproteins / Q000276:immunology; D051379:Mice; D016688:Mice, Inbred NOD; D021381:Protein Transport; D017452:Receptors, IgG / Q000378:metabolism; D014161:Transduction, Genetic; D014759:Viral Envelope Proteins / Q000276:immunology
D057134:Antibodies, Neutralizing; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins; D017452:Receptors, IgG; D014759:Viral Envelope Proteins