Two protection-inducing antigens specific to the L1 larva of Trichinella spiralis will be investigated regarding their structure and function using immunological, biochemical and molecular biological approaches. Expression and genomic libraries in suitable vectors will be constructed and cloned DNA molecules encoding both the 48K and 50/55K protein will be isolated. Expression of positive cDNA clones will be identified by colony immune assay with polyvalent rabbit anti-48K and 50/55K antibodies. Fusion peptides containing epitopes will be tested for their ability to induce protection in mice. Southern blot analysis will determine the genomic arrangement of DNA encoding both antigens. Northern analysis on each stage of Trichinella spiralis (L1-L4 larva, adult and newborn larva) will reveal the stage- specificity of gene expression for each antigen. The sequence of cDNA and genomic DNA for each antigen will be determined. Both proteins can be phosphorylated, and in addition, the 48K antigen binds divalent cations, particularly Ca2+. Thus peptide mapping and sequencing studies on each molecule will be carried out to determine the sites of phosphorylation for both proteins, and the number and sites of Ca2+ binding for the 48K molecule. Computer analysis of the primary amino acid consensus sequence and DNA sequence will determine areas of hydrophobicity and degrees of homology with other known sequences. Particular attention to similarities between regulatory proteins and the 48K and 50/55K antigens will be given, since preliminary data indicate that they may be involved in regulation of host anerobic glycolysis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI010627-19
Application #
3124752
Study Section
Tropical Medicine and Parasitology Study Section (TMP)
Project Start
1977-06-01
Project End
1991-05-31
Budget Start
1989-06-01
Budget End
1990-05-31
Support Year
19
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Public Health
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
Polvere, R I; Kabbash, C A; Capo, V A et al. (1997) Trichinella spiralis: synthesis of type IV and type VI collagen during nurse cell formation. Exp Parasitol 86:191-9
Vassilatis, D K; Despommier, D; Misek, D E et al. (1992) Analysis of a 43-kDa glycoprotein from the intracellular parasitic nematode Trichinella spiralis. J Biol Chem 267:18459-65
Gold, A M; Despommier, D D; Buck, S W (1990) Partial characterization of two antigens secreted by L1 larvae of Trichinella spiralis. Mol Biochem Parasitol 41:187-96
Despommier, D D; Gold, A M; Buck, S W et al. (1990) Trichinella spiralis: secreted antigen of the infective L1 larva localizes to the cytoplasm and nucleoplasm of infected host cells. Exp Parasitol 71:27-38