A variety of different mechanisms to combat pathogen challenges have emerged during metazoan evolution. The diverse nature of form, function and phylogenetic integration of these processes is significant for understanding basic mechanisms governing immune recognition. Acquisition of complete genome sequences in species that represent key points in phylogeny not only permit in-depth analysis of immune mediators but also represent an invaluable resource for understanding genetic mechanisms that create and maintain polymorphism and haplotype complexity. Such information is critical for gaining a more comprehensive view of: interrelationships within the innate immune system, the basis for complex polymorphisms in certain innate receptors and the origins of adaptive immunity. Amphioxus (a cephalochordate) occupies a unique phylogenetic position that represents the base of chordate evolution. Two striking features distinguish immune- related function in this species: 1) large-scale expansions and diversification have occurred in gene families encoding Toll-like receptor (TLR) innate immune receptors as well as other immune gene families and 2) a multigene family encoding immunoglobulin variable domain-containing chitin-binding proteins (VCBPs) exhibits an extraordinary level of regionalized hypervariation. In order to understand the genetics and functions of these phenomena, monomeric and multivalent, chimeric VCBP and TLR immune-type receptor genes will be engineered. In order to characterize ligands and ligand binding, conventional screening methods, metagenomic approaches and phage display will be adapted. Responses to immune stimulation will be characterized in cell types expressing VCBPs, TLR-related molecules and other innate receptors. Cellular specialization will be related to receptor diversity and mechanisms that sense pathogen signals. Transcriptional complexity of immune-related molecules will be addressed using a novel BAC transfection-transcriptome readout approach. Haplotypic variation will be characterized and germline as well as potential somatic variations will be examined. The mechanisms underlying the genetic variation seen in VCBPs at the natural population level will be examined by characterizing individual gametes. The relationships of genomic context, specifically repetitive DNA and noncoding haplotypic differences, to the germline complexity of VCBPs will be examined through direct study of meiotic variants as well as by plasmid- and BAC-transgenesis in zebrafish as a high throughput readout system. Collectively, a more comprehensive understanding of innate receptor diversification and function as well as relationships between polymorphism, functional variation and their relationship to chromosomal context will emerge. Such information is of particular relevance to current concepts of host responses to pathogen challenge and mechanisms used to create and maintain functional polymorphism of immune receptors.

Public Health Relevance

A remarkable degree of complexity and variability in immune mechanisms has occurred during evolution and likely reflects major interspecies differences in host-pathogen interactions. Amphioxus (Branchiostoma floridae) is representative of an early point in the development of the chordate form, which includes humans. The genome of amphioxus has been resolved and two striking features are evident: 1) genes encoding many innate immune receptors, which provide the first line of defense, are 10-fold increased in number than are seen in mammals and 2) an immunoglobulin variable region-containing immune-type receptor exhibits regionalized hypervariation that is associated with unique chromosomal features. We seek to examine the nature of the variation in these receptors, relate genetic diversity to specific cell lineage(s) and examine the relationships between receptor variation and genome context as a means to gain a more basic understanding of the complex roles of immune receptors in combating disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI023338-29
Application #
8580545
Study Section
Cellular and Molecular Immunology - B Study Section (CMIB)
Program Officer
Rice, Jeffrey S
Project Start
1985-09-01
Project End
2014-11-30
Budget Start
2013-12-01
Budget End
2014-11-30
Support Year
29
Fiscal Year
2014
Total Cost
$327,443
Indirect Cost
$104,693
Name
University of South Florida
Department
Pediatrics
Type
Schools of Medicine
DUNS #
069687242
City
Tampa
State
FL
Country
United States
Zip Code
33612
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Zhang, Linlin; Li, Li; Guo, Ximing et al. (2015) Massive expansion and functional divergence of innate immune genes in a protostome. Sci Rep 5:8693
Rodríguez-Nunez, Iván; Wcisel, Dustin J; Litman, Gary W et al. (2014) Multigene families of immunoglobulin domain-containing innate immune receptors in zebrafish: deciphering the differences. Dev Comp Immunol 46:24-34
Saha, Nil Ratan; Ota, Tatsuya; Litman, Gary W et al. (2014) Genome complexity in the coelacanth is reflected in its adaptive immune system. J Exp Zool B Mol Dev Evol 322:438-63
Liberti, Assunta; Melillo, Daniela; Zucchetti, Ivana et al. (2014) Expression of Ciona intestinalis variable region-containing chitin-binding proteins during development of the gastrointestinal tract and their role in host-microbe interactions. PLoS One 9:e94984
Kortum, Amanda N; Rodriguez-Nunez, Ivan; Yang, Jibing et al. (2014) Differential expression and ligand binding indicate alternative functions for zebrafish polymeric immunoglobulin receptor (pIgR) and a family of pIgR-like (PIGRL) proteins. Immunogenetics 66:267-79
Dishaw, Larry J; Flores-Torres, Jaime; Lax, Simon et al. (2014) The gut of geographically disparate Ciona intestinalis harbors a core microbiota. PLoS One 9:e93386
Dishaw, Larry J; Cannon, John P; Litman, Gary W et al. (2014) Immune-directed support of rich microbial communities in the gut has ancient roots. Dev Comp Immunol 47:36-51
Cannon, John P; O'Driscoll, Marci; Litman, Gary W (2012) Specific lipid recognition is a general feature of CD300 and TREM molecules. Immunogenetics 64:39-47
Haire, Robert N; Cannon, John P; O'Driscoll, Marci L et al. (2012) Genomic and functional characterization of the diverse immunoglobulin domain-containing protein (DICP) family. Genomics 99:282-91

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