We propose to use our recent discovery that the forced expression of AID in hybridomas activates the somatic V region mutation to generate more effective monoclonal antibodies to agents used in bioterrorism. We will test high rates of V region mutation and isotype switching that can be activated in vivo with the conditional expression of AID. We will establish the optimum conditions for multiple rounds of mutation and selection to obtain the highest affinity antibodies with the full spectrum of isotypes. We will adapt techniques that we have used previously and we will develop new ones that will allow us to identify hybridoma subclones that are making human and mouse monoclonal antibodies that will more effectively neutralize the toxins and viruses and protect against microbial agents that are the primary agents used in bioterrorism and germ warfare. We will determine if such second-generation monoclonal antibodies to ricin and other toxins are truly more effective using mouse models.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21AI053362-01
Application #
6562150
Study Section
Special Emphasis Panel (ZAI1-GPJ-M (M3))
Program Officer
Nasseri, M Faraz
Project Start
2002-09-01
Project End
2004-08-31
Budget Start
2002-09-01
Budget End
2003-08-31
Support Year
1
Fiscal Year
2002
Total Cost
$250,500
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461
Iglesias-Ussel, Maria D; Fan, Manxia; Li, Ziqiang et al. (2006) Forced expression of AID facilitates the isolation of class switch variants from hybridoma cells. J Immunol Methods 316:59-66
Ronai, Diana; Iglesias-Ussel, Maria Dolores; Fan, Manxia et al. (2005) Complex regulation of somatic hypermutation by cis-acting sequences in the endogenous IgH gene in hybridoma cells. Proc Natl Acad Sci U S A 102:11829-34
Bardwell, Philip D; Woo, Caroline J; Wei, Kaichun et al. (2004) Altered somatic hypermutation and reduced class-switch recombination in exonuclease 1-mutant mice. Nat Immunol 5:224-9
Woo, Caroline J; Martin, Alberto; Scharff, Matthew D (2003) Induction of somatic hypermutation is associated with modifications in immunoglobulin variable region chromatin. Immunity 19:479-89
Bransteitter, Ronda; Pham, Phuong; Scharff, Matthew D et al. (2003) Activation-induced cytidine deaminase deaminates deoxycytidine on single-stranded DNA but requires the action of RNase. Proc Natl Acad Sci U S A 100:4102-7