Cloning and expression of the Actinobacillus actinomycetemcomitans thioredoxin (trx) gene and assessment of cytokine inhibitory activity.
Thioredoxin is a ubiquitous redox control and cell stress protein. Unexpectedly, in recent years, thioredoxins have been found to exhibit both cytokine and chemokine activities, and there is increasing evidence that this class of protein plays a role in the pathogenesis of inflammatory diseases. In spite of this evidence, it has been reported that the oral bacterium and periodontopathogen Actinobacillus actinomycetemcomitans secretes an immunosuppressive factor (termed suppressive factor 1 [SF1] [T. Kurita-Ochiai and K. Ochiai, Infect. Immun. 64:50-54, 1996]) whose N-terminal sequence, we have determined, identifies it as thioredoxin. We have cloned and expressed the gene encoding the thioredoxin of A. actinomycetemcomitans and have purified the protein to homogeneity. The A. actinomycetemcomitans trx gene has 52 and 76% identities, respectively, to the trx genes of Escherichia coli and Haemophilus influenzae. Enzymatic analysis revealed that the recombinant protein had the expected redox activity. When the recombinant thioredoxin was tested for its capacity to inhibit the production of cytokines by human peripheral blood mononuclear cells, it showed no significant inhibitory capacity. We therefore conclude that the thioredoxin of A. actinomycetemcomitans does not act as an immunosuppressive factor, at least with human leukocytes in cultures, and that the identity of SF1 remains to be elucidated.
Henderson
B
B
Cellular Microbiology Research Group, Eastman Dental Institute, University College London, London WC1X 8LD, United Kingdom. b.henderson@eastman.ucl.ac.uk
Tabona
P
P
Poole
S
S
Nair
S P
SP
D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics*; D000595:Amino Acid Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D016207:Cytokines / Q000037:antagonists & inhibitors*; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017510:Protein Folding; D013879:Thioredoxins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology
D016207:Cytokines; D007166:Immunosuppressive Agents; D013879:Thioredoxins