Generation of regulatory gut-homing human T lymphocytes using ex vivo interleukin 10 gene transfer.
OBJECTIVE
Systemic treatment of Crohn's disease patients using recombinant interleukin (rIL)-10 has not resulted in significant therapeutic benefit presumably because of limited bioavailability and unexpected proinflammatory effects of high-dose rIL-10. Ex vivo gene transfer of the interleukin (IL)-10 gene to gut-homing CD4(+) cells may lead to improved long-term management.


METHODS
Peripheral blood mononuclear cells (PBMCs) were transduced with a retroviral vector containing the IL-10 and green fluorescent protein (GFP) gene or a control vector containing GFP only. Transduced CD4(+) cells were sorted and maintained in culture for phenotypic and functional analysis.


RESULTS
Stimulated IL-10-GFP CD4(+) cells produced significantly higher levels of IL-10 than control cells for at least 4 months. The IL-10 transgene was biologically active and decreased proliferation of IL-10-GFP CD4(+) cells as well as expression of major histocompatibility class (MHC) class II, proliferation of autologous responder cells, and IL-12 production by dendritic cells (DCs). The majority of transduced CD4(+) cells had a gut-homing potential because they expressed the mucosal integrin alpha4beta7, and displayed efficient binding to MAdCAM-1-expressing cells in vitro.


CONCLUSIONS
Transduction of peripheral blood CD4(+) lymphocytes with IL-10 results in a regulatory phenotype. The use of regulatory gut-homing human CD4(+) cells may provide a novel approach to local delivery of immunomodulatory signals to the intestine in Crohn's disease.
Van Montfrans
Catherine
C
Department of Experimental Internal Medicine, Academic Medical Center, Amsterdam, The Netherlands.
Hooijberg
Erik
E
Rodriguez Pena
Maria Sol
MS
De Jong
Esther C
EC
Spits
Hergen
H
Te Velde
Anje A
AA
Van Deventer
Sander J H
SJ
D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000378:metabolism; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D006801:Humans; D007136:Immunoglobulins / Q000502:physiology; D016753:Interleukin-10 / Q000235:genetics*; D018664:Interleukin-12 / Q000096:biosynthesis; D007422:Intestines / Q000502:physiology*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D009088:Mucoproteins / Q000502:physiology; D019899:Paracrine Communication / Q000502:physiology; D016201:Receptors, Lymphocyte Homing / Q000502:physiology*; D013601:T-Lymphocytes / Q000502:physiology*; D014161:Transduction, Genetic
D015815:Cell Adhesion Molecules; D016207:Cytokines; D007136:Immunoglobulins; D008164:Luminescent Proteins; C504265:MADCAM1 protein, human; D009088:Mucoproteins; D016201:Receptors, Lymphocyte Homing; D016753:Interleukin-10; D049452:Green Fluorescent Proteins; D018664:Interleukin-12