|Smith, Timothy - Tim|
Submitted to: Advances in Genome Biology and Technology
Publication Type: Abstract Only
Publication Acceptance Date: 12/30/2012
Publication Date: 2/20/2013
Citation: Smith, T.P.L., and Larsen, P.A. 2013. Deep sequencing of immune repertoires during bovine development and in response to respiratory pathogen challenge [Abstract]. Advances in Genome Biology and Technology Conference, February 20-23, 2013, Marco Island, FL. Page 126.
Technical Abstract: Vertebrate immune systems generate diverse repertoires of antibodies capable of mediating response to a variety of antigens. Single-molecule circular consensus sequencing permits the sequencing of expressed antibody repertoires at previously unattainable depths of coverage and accuracy. We examined the bovine immunoglobulin G (IgG) repertoire with the objective of characterizing diversity of expressed IgG transcripts during postnatal development and in response to immunological insult in the form of pathogen challenge. We focused on expressed transcripts because our primary interest is in detecting repertoire changes among activated cells, and because a germ line sequence for the cattle immunoglobulin locus is not available for design of genome-based primers. Using a network-based approach to characterize complementarity-determining region (CDR) diversity, we observed changes in repertoires of eleven animals as they progressed from neonate to weaning, during weekly samplings over the first twelve weeks after birth. Immediately after birth, circulating lymphocytes were too low in number to support robust determination of CDR repertoires because cattle are passively immunized through colostrum early in life; sufficient small lymphocytes were not observed until the third week after birth. Rapid proliferation of the potential antigen binding repertoire was observed during subsequent development, and comparative aspects among CDR of individual animals will be presented. In addition, in preliminary experiments we observed profound alterations in repertoire in animals presented with pathogenic challenge, supporting the hypothesis that immune repertoire characterization can provide insights into disease processes and the health status of individuals, as well as potentially guiding antibody discovery and engineering, disease surveillance, and host immune response to vaccines.