In the current grant we propose to expand our studies of genetically engineered antibody molecules, focusing our attention on the goal of providing more effective immunity to the fetus and neonate. In one series of experiments we will identify and produce antibodies with the optimal combination of functional properties to provide protection at the mucosal surface following oral administration or transport from the mother via the placenta or mammary gland. Human IgA is the antibody normally found in the exocrine secretions. IgA, domain exchange proteins between human IgA and IgG, and site directed mutants will be characterized with respect to their stability, half-life, and biodistribution when administered either orally or intravenously. They will be characterized with respect to their ability to bind and be transported by the epithelial receptors. They also will be assessed for their ability to activate the complement cascade and to bind to Fc receptors specific for either IgG or IgA on phagocytes. We will also attempt to develop a cell which expresses antibody with attached secretory component (SC). One goal will be to produce recombinant antibodies which provide treatment or protection against enteric infections, a significant health problem world wide. We will assay the most promising recombinant antibodies in the suckling mouse model for enterotoxigenic Escherichia coli (ETEC) using antibodies specific for the F41 antigen. Antibodies to F4l have proven efficacy and recombinant antibodies administered orally to pups or intravenously to dams will be assessed for their ability to prevent lethality and to decrease colonization. In vitro assays of adherence, agglutination and complement mediated cytotoxicity will address the mechanism(s) of protection. If recombinant antibodies are to be broadly applied, an inexpensive expression system must be available and we will focus our attention on the development of the transgenic chicken as such an expression system. The chicken is inexpensive to maintain and targets large amounts of antibody to the egg yolk. Intravenous injection of iodinated antibodies into laying hens will be used to determine which antibodies will be transported to the yolk. Vectors tested for Ig expression using chicken lymphoid cell lines will be transfected into Stage X blastoderm cells and these transfected cells used to make chimeric embryos. The resulting chickens will be tested for the presence of chimeric lg in their serum and in the eggs of laying hens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI039187-04
Application #
2672673
Study Section
Special Emphasis Panel (SRC (41))
Project Start
1995-09-30
Project End
2000-08-31
Budget Start
1998-09-01
Budget End
1999-08-31
Support Year
4
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Chintalacharuvu, Koteswara R; Gurbaxani, Brian; Morrison, Sherie L (2007) Incomplete assembly of IgA2m(2) in Chinese hamster ovary cells. Mol Immunol 44:3445-52
Gurbaxani, Brian Mohan; Morrison, Sherie L (2006) Development of new models for the analysis of Fc-FcRn interactions. Mol Immunol 43:1379-89
Dela Cruz, Jay Soriano; Trinh, Kamh Ryan; Chen, Hsiao Wen et al. (2006) Anti-HER2/neu IgG3-(IL-2) and anti-HER2/neu IgG3-(GM-CSF) promote HER2/neu processing and presentation by dendritic cells: implications in immunotherapy and vaccination strategies. Mol Immunol 43:667-76
Helguera, Gustavo; Dela Cruz, Jay S; Lowe, Christine et al. (2006) Vaccination with novel combinations of anti-HER2/neu cytokines fusion proteins and soluble protein antigen elicits a protective immune response against HER2/neu expressing tumors. Vaccine 24:304-16
Gurbaxani, Brian; Dela Cruz, Linh L; Chintalacharuvu, Koteswara et al. (2006) Analysis of a family of antibodies with different half-lives in mice fails to find a correlation between affinity for FcRn and serum half-life. Mol Immunol 43:1462-73
Zhu, Lei; van de Lavoir, Marie-Cecile; Albanese, Jenny et al. (2005) Production of human monoclonal antibody in eggs of chimeric chickens. Nat Biotechnol 23:1159-69
Dela Cruz, Jay S; Morrison, Sherie L; Penichet, Manuel L (2005) Insights into the mechanism of anti-tumor immunity in mice vaccinated with the human HER2/neu extracellular domain plus anti-HER2/neu IgG3-(IL-2) or anti-HER2/neu IgG3-(GM-CSF) fusion protein. Vaccine 23:4793-803
Trinh, Ryan; Gurbaxani, Brian; Morrison, Sherie L et al. (2004) Optimization of codon pair use within the (GGGGS)3 linker sequence results in enhanced protein expression. Mol Immunol 40:717-22
Gala, Francoise A; Morrison, Sherie L (2004) V region carbohydrate and antibody expression. J Immunol 172:5489-94
Chan, Lisa A; Phillips, Martin L; Wims, Letitia A et al. (2004) Variable region domain exchange in human IgGs promotes antibody complex formation with accompanying structural changes and altered effector functions. Mol Immunol 41:527-38

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