The expression and secretion of the recombinant proteins were confirmed in vitro using immunofluorescence (IF) and western blot (WB) analyses. Human dengue immune serum samples recognized recombinant proteins. The immunogenicity of the 4 proteins in BALB/c mice was analyzed using ELISA, and the results revealed that the induced specific antibody response was higher in the groups of mice immunized with the P28 fusion proteins. Interestingly, although the 4 recombinant proteins selleckchem were able to elicit high levels of
neutralizing antibodies in BALB/c mice; no adjuvant effect was observed in terms of neutralizing antibodies in the groups immunized with proteins containing P28. Thus, ELISA and PRNT50 assays may evaluate different epitopes
and responses, where ELISA showed a wider response that did not always correlate with neutralization. Furthermore, the elicited antibodies were able to recognize the immobilized E glycoprotein of DENV. All mice vaccinated with the DENV-2 recombinant proteins showed induction of higher levels of IgG1 antibodies than of IgG2a antibodies.”
“Objective: To compare the efficacy of in situ transforming growth factor-betal (TGF-beta 1)-pretreated periosteum to untreated periosteum for regeneration of osteochondral tissue in rabbits.
Methods: In the pretreatment group, 12 month-old New Zealand white rabbits received subperiosteal Milciclib injections of 200 ng of TGF-beta 1 percutaneously in the medial side of the proximal tibia, 7 days prior to surgery. Control rabbits received no treatment prior surgery. Osteochondral transverse defects measuring 5 mm proximal to distal and spanning the entire width of the patellar groove BLZ945 were created and repaired with untreated or TGF-beta 1-pretreated periosteal grafts. Post-operatively the rabbits resumed normal cage activity for 6 weeks.
Results: Complete filling of the defects with regenerated
tissue was observed in both the TGF-beta 1 -pretreated and control groups with reformation of the original contours of the patellar groove. The total histological score (modified O’Driscoll) in the TGF-beta 1-pretreated group, 20(95% Confidence Interval (CI), 19-21), was significantly higher (P=0.0001) than the control group, 18 (16-19). The most notable improvements were in structural integrity and subchondral bone regeneration. No significant differences in glycosaminoglycan or type II collagen content, or equilibrium modulus were found between the surgical groups. The cambium of the periosteum regenerated at the graft harvest site was significantly thicker (P=0.0065) in the TGF-beta 1-pretreated rabbits, 121 mu m (94-149), compared to controls, 74 mu m (52-96), after 6 weeks.
Conclusions: This study demonstrates that in situ pretreatment of periosteum with TGF-beta 1 improves osteochondral tissue regeneration at 6-weeks post-op compared to untreated periosteum in 12 month-old rabbits. (C) 2010 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.