A human anti-Pseudomonas aeruginosa serotype O6ad immunoglobulin G1 expressed in transgenic tobacco is capable of recruiting immune system effector function in vitro.
The production of a recombinant human IgG1 in transgenic tobacco was examined to determine whether a plant-derived antibody could recruit immune system effector function against a bacterial pathogen. A plant transformation vector was engineered to contain genes for a human kappa light chain and a human gamma-1 heavy chain with V(H) and V(L) sequences from a previously identified human IgG2 monoclonal antibody (MAb) that specifically binds to and opsonizes Pseudomonas aeruginosa serotype O6ad. Unique NcoI and NotI restriction sites were incorporated to flank these variable sequences, resulting in a plant transformation vector that could be engineered for expression of any other human IgG1 antibody, requiring only the substitution of other V(H) and V(L) antigen-binding coding sequences. The plant-produced IgG1 was determined to have high-mannose glycan content and to be capable of mediating opsonophagocytosis of P. aeruginosa serotype O6ad in vitro using human complement and human polymorphonuclear leukocytes. Thus, MAbs produced in plants from this vector could provide human IgG1 MAbs for targeting other pathogens that require the recruitment of immune system effector functions.
McLean
Michael D
MD
Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1 Canada.
Almquist
Kurt C
KC
Niu
Yongfing
Y
Kimmel
Rhonda
R
Lai
Zengzu
Z
Schreiber
John R
JR
Hall
J Christopher
JC
D000911:Antibodies, Monoclonal / Q000737:chemistry; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D007107:Immune System / Q000187:drug effects* / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology*; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000276:immunology; D066298:In Vitro Techniques; D008358:Mannose / Q000378:metabolism; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000187:drug effects; D009895:Opsonin Proteins / Q000494:pharmacology; D010587:Phagocytosis / Q000187:drug effects; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011134:Polysaccharides / Q000276:immunology / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D014026:Nicotiana / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D009895:Opsonin Proteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D008358:Mannose