Oral Tolerance Induction in Hemophilia B Dogs Fed with Transplastomic Lettuce.
Anti-drug antibodies in hemophilia patients substantially complicate treatment. Their elimination through immune tolerance induction (ITI) protocols poses enormous costs, and ITI is often ineffective for factor IX (FIX) inhibitors. Moreover, there is no prophylactic ITI protocol to prevent anti-drug antibody (ADA) formation. Using general immune suppression is problematic. To address this urgent unmet medical need, we delivered antigen bioencapsulated in plant cells to hemophilia B dogs. Commercial-scale production of CTB-FIX fusion expressed in lettuce chloroplasts was done in a hydroponic facility. CTB-FIX (∼1 mg/g) in lyophilized cells was stable with proper folding, disulfide bonds, and pentamer assembly after 30-month storage at ambient temperature. Robust suppression of immunoglobulin G (IgG)/inhibitor and IgE formation against intravenous FIX was observed in three of four hemophilia B dogs fed with lyophilized lettuce cells expressing CTB-FIX. No side effects were detected after feeding CTB-FIX-lyophilized plant cells for >300 days. Coagulation times were markedly shortened by intravenous FIX in orally tolerized treated dogs, in contrast to control dogs that formed high-titer antibodies to FIX. Commercial-scale production, stability, prolonged storage of lyophilized cells, and efficacy in tolerance induction in a large, non-rodent model of human disease offer a novel concept for oral tolerance and low-cost production and delivery of biopharmaceuticals.
Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
Roland W
RW
Herzog
Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, Chapel Hill, NC 25716, USA.
Timothy C
TC
Nichols
Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Jin
J
Su
Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Bei
B
Zhang
Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
Alexandra
A
Sherman
Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, Chapel Hill, NC 25716, USA.
Elizabeth P
EP
Merricks
Department of Pathology and Laboratory Medicine, The University of North Carolina, Chapel Hill, Chapel Hill, NC 25716, USA.
Robin
R
Raymer
Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
George Q
GQ
Perrin
Global Research, Novo Nordisk A/S, Måløv 2760, Denmark.
Mattias
M
Häger
Global Research, Novo Nordisk A/S, Måløv 2760, Denmark.
Bo
B
Wiinberg
Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: hdaniell@upenn.edu.
Henry
H
Daniell
D000284:Administration, Oral; D000821:Animal Feed; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000917:Antibody Formation; D015415:Biomarkers; D002736:Chloroplasts / Q000235:genetics; D004285:Dogs; D005164:Factor IX / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D002836:Hemophilia B / Q000097:blood / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007108:Immune Tolerance; D018545:Lactuca / Q000235:genetics; D010641:Phenotype; D030821:Plants, Genetically Modified
D000906:Antibodies; D015415:Biomarkers; D005164:Factor IX
factor IX; hemophilia; inhibitor; oral tolerance; transgenic plant