Innate and adaptive immunity to human beta cell lines:  implications for beta cell therapy.
OBJECTIVE
Genetically engineered human beta cell lines provide a novel source of human beta cells to study metabolism, pharmacology and beta cell replacement therapy. Since the immune system is essentially involved in beta cell destruction in type 1 diabetes and after beta cell transplantation, we investigated the interaction of human beta cell lineswith the immune system to resolve their potential for immune intervention protocol studies.


METHODS
Human pancreatic beta cell lines (EndoC-βH1 and ECi50) generated by targeted oncogenesis in fetal pancreas were assessed for viability after innate and adaptive immune challenges. Beta cell lines were pre-conditioned with T helper type 1 (Th1) cytokines or high glucose to mimic inflammatory and hyperglycaemia-stressed conditions. Beta cells were then co-cultured with auto- and alloreactive cytotoxic T cells (CTL), natural killer (NK) cells, supernatant fraction from activated autoreactive Th1 cells, or alloantibodies in the presence of complement or effector cells.


RESULTS
Low HLA expression protected human beta cell lines from adaptive immune destruction, but it was associated with direct killing by activated NK cells. Autoreactive Th1 cell inflammation, rather than glucose stress, induced increased beta cell apoptosis and upregulation of HLA, increasing beta cell vulnerability to killing by auto- and alloreactive CTL and alloreactive antibodies.


CONCLUSIONS
We demonstrate that genetically engineered human beta cell lines can be used in vitro to assess diverse immune responses that may be involved in the pathogenesis of type 1 diabetes in humans and beta cell transplantation, enabling preclinical evaluation of novel immune intervention strategies protecting beta cells from immune destruction.
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, E3-Q, P.O. Box 9600, 2300 RC, Leiden, the Netherlands.
Cornelis R
CR
van der Torren
Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, the Netherlands.
Arnaud
A
Zaldumbide
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, E3-Q, P.O. Box 9600, 2300 RC, Leiden, the Netherlands.
Dave L
DL
Roelen
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, E3-Q, P.O. Box 9600, 2300 RC, Leiden, the Netherlands.
Gaby
G
Duinkerken
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, E3-Q, P.O. Box 9600, 2300 RC, Leiden, the Netherlands.
Simone H
SH
Brand-Schaaf
Department of Immunobiology, School of Medicine, King's College London, London, UK.
Mark
M
Peakman
Endocells, Paris, France.
Paul
P
Czernichow
CNRS UMR 7725, Université Pierre et Marie Curie (UPMC), Paris, France.
Philippe
P
Ravassard
Inserm U1016, Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
Raphael
R
Scharfmann
Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, E3-Q, P.O. Box 9600, 2300 RC, Leiden, the Netherlands. boroep@lumc.nl.
Bart O
BO
Roep
D056704:Adaptive Immunity; D000906:Antibodies / Q000276:immunology; D002460:Cell Line; D017690:Cell Transplantation / Q000379:methods; D003165:Complement System Proteins / Q000276:immunology; D016207:Cytokines / Q000378:metabolism; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology; D005818:Genetic Engineering / Q000379:methods; D005838:Genotype; D006680:HLA Antigens / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D006943:Hyperglycemia / Q000378:metabolism; D007107:Immune System; D007113:Immunity, Innate; D007249:Inflammation; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000276:immunology*; D007694:Killer Cells, Natural / Q000166:cytology; D007963:Leukocytes, Mononuclear / Q000166:cytology; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology; D018417:Th1 Cells / Q000166:cytology
D000906:Antibodies; D016207:Cytokines; D006680:HLA Antigens; D003165:Complement System Proteins