A minimized Fc binding peptide from protein A induces immunocyte proliferation and evokes Th1-type response in mice.
It is now well established that PA is a potent biological response modifier, showing simultaneously antitumor, antitoxic, anticarcinogenic, antifungal, antiparasitic and immunomodulatory properties. Since PA is a foreign protein, it is quite logical to assume that it may be cleaved into smaller peptide fragments in vivo which may be responsible for biological activities of whole PA molecule. The present study was undertaken to dissect out the structural entities of PA responsible for its biological properties. Protein A (PA) of Staphylococcus aureus has a unique property of binding with immunoglobulins. On the basis of molecular modeling and energy minimization studies a 20-mer tryptic fragment (theoretical) was predicted to retain IgG binding capacity which has been verified by immunoblot. This peptide sequence was selected to carry out experimental studies to show its functional mimicry of PA. We observed in the sera of 20-mer peptide treated mice that the concentrations of IFNgamma, TNFalpha and IL1alpha increase to a peak level by 4 h; on the other hand, there was a decrease in IL4, IL6 and IL10 concentrations at the same time (4 h). The ratio of IFNgamma to IL4 showed Th1 type of response with the peptide as well as with that of PA. The nitric oxide concentration in sera also increases and the peak increase was in 6 h with both the peptide and PA. Cell cycle analysis using FACS shows that 20 micrograms dose of peptide was non-toxic to thymocytes and spleenocytes; on the other hand, it was immunoproliferative, shifting the thymocytes and spleenocytes from G0/G1 to S phase of the cell cycle. Further studies are in progress to evaluate other biological properties of the peptide, to evaluate if this peptide could be used as a substitute of PA to mimic at least some of its biological activities.
Sinha
P
P
Immunotechnology Section, Bose Institute, Calcutta, 700 054, India.
Sengupta
J
J
Ray
P K
PK
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002453:Cell Cycle; D002455:Cell Division / Q000502:physiology*; D016207:Cytokines / Q000097:blood; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism*; D008297:Male; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000096:biosynthesis; D010446:Peptide Fragments / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding; D013205:Staphylococcal Protein A / Q000737:chemistry*; D018417:Th1 Cells / Q000166:cytology / Q000276:immunology*
D016207:Cytokines; D007141:Immunoglobulin Fc Fragments; D010446:Peptide Fragments; D013205:Staphylococcal Protein A; D009569:Nitric Oxide