Transcriptomic basis for an antiserum against Micrurus corallinus (coral snake) venom.
BACKGROUND
Micrurus corallinus (coral snake) is a tropical forest snake belonging to the family Elapidae. Its venom shows a high neurotoxicity associated with pre- and post-synaptic toxins, causing diaphragm paralysis, which may result in death. In spite of a relatively small incidence of accidents, serum therapy is crucial for those bitten. However, the adequate production of antiserum is hampered by the difficulty in obtaining sufficient amounts of venom from a small snake with demanding breeding conditions. In order to elucidate the molecular basis of this venom and to uncover possible immunogens for an antiserum, we generated expressed sequences tags (ESTs) from its venom glands and analyzed the transcriptomic profile. In addition, their immunogenicity was tested using DNA immunization.


RESULTS
A total of 1438 ESTs were generated and grouped into 611 clusters. Toxin transcripts represented 46% of the total ESTs. The two main toxin classes consisted of three-finger toxins (3FTx) (24%) and phospholipases A(2) (PLA(2)s) (15%). However, 8 other classes of toxins were present, including C-type lectins, natriuretic peptide precursors and even high-molecular mass components such as metalloproteases and L-amino acid oxidases. Each class included an assortment of isoforms, some showing evidence of alternative splicing and domain deletions. Five antigenic candidates were selected (four 3FTx and one PLA(2)) and used for a preliminary study of DNA immunization. The immunological response showed that the sera from the immunized animals were able to recognize the recombinant antigens.


CONCLUSIONS
Besides an improvement in our knowledge of the composition of coral snake venoms, which are very poorly known when compared to Old World elapids, the expression profile suggests abundant and diversified components that may be used in future antiserum formulation. As recombinant production of venom antigens frequently fails due to complex disulfide arrangements, DNA immunization may be a viable alternative. In fact, the selected candidates provided an initial evidence of the feasibility of this approach, which is less costly and not dependent on the availability of the venom.
Leão
Luciana I
LI
Centro de Biotecnologia, Instituto Butantan, São Paulo, SP, Brazil. lucianaleao@butantan.gov.br
Ho
Paulo L
PL
Junqueira-de-Azevedo
Inacio de L M
Ide L
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000276:immunology; D000997:Antivenins; D016000:Cluster Analysis; D004546:Elapid Venoms / Q000235:genetics* / Q000276:immunology; D017815:Elapidae / Q000235:genetics*; D020224:Expressed Sequence Tags; D005260:Female; D020869:Gene Expression Profiling; D015723:Gene Library; D007106:Immune Sera / Q000032:analysis; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D019444:Vaccines, DNA / Q000276:immunology
D000941:Antigens; D000997:Antivenins; D004546:Elapid Venoms; D007106:Immune Sera; D007074:Immunoglobulin G; D020033:Protein Isoforms; D019444:Vaccines, DNA