BIOPHYSICAL AND IMMUNOLOGICAL STUDIES WITH A RECOMBINANT PLASMODIUM FALCIPARUM CIRCUMSPOROZOITE PROTEIN (PFCSP) Jennifer Carter1, Christopher Pool1, Brooke A. Bozick1, Particia De la Vega1, Evelina Angov1, Roberta Spaccapelo2, Andrea Crisanti3, Jittawadee R. Murphy1, Christian F. Ockenhouse1, Sheetij Dutta1 1Walter Reed Army Institute of Research, Silver Spring, MD, United States, 2Università degli Studi di Perugia, Perugia, Italy, 3Imperial College London, London, United Kingdom A highly purified preparation of a near full length recombinant circumsporozoite protein (PfCSP) was analyzed by SDS-PAGE and shown to migrate ~10 kDa higher than its predicted 30 kDa molecular weight under both reducing and non-reducing conditions. Blue-native non-denaturing PAGE and analytical size exclusion chromatography further confirmed that the rCSP monomer had a highly extended molecular structure and its size was equivalent to a ~60 kDa globular protein. Vaccination of PfCSP along with an adjuvant Montanide ISA720 induced high titer antibodies in mice and this vaccination conferred sterile protection in two strains of mice against challenge with a transgenic Plasmodium berghei parasite line that expressed the P. falciparum CSP gene. The anti-PfCSP antibodies recognized the native CSP on sporozoites by IFA and inhibited the invasion of NF54 strain sporozoites into hepatocytes. Availability of a high quality near full-length rCSP and the transgenic mouse protection model will allow us to develop novel strategies to enhance the protective efficacy of CSP based vaccines in humans.
BIOPHYSICAL AND IMMUNOLOGICAL STUDIESWITH A RECOMBINANT PLASMODIUM FALCIPARUMCIRCUMSPOROZOITE PROTEIN (PFCSP)
SPACCAPELO, Roberta;CRISANTI, Andrea;
2010
Abstract
BIOPHYSICAL AND IMMUNOLOGICAL STUDIES WITH A RECOMBINANT PLASMODIUM FALCIPARUM CIRCUMSPOROZOITE PROTEIN (PFCSP) Jennifer Carter1, Christopher Pool1, Brooke A. Bozick1, Particia De la Vega1, Evelina Angov1, Roberta Spaccapelo2, Andrea Crisanti3, Jittawadee R. Murphy1, Christian F. Ockenhouse1, Sheetij Dutta1 1Walter Reed Army Institute of Research, Silver Spring, MD, United States, 2Università degli Studi di Perugia, Perugia, Italy, 3Imperial College London, London, United Kingdom A highly purified preparation of a near full length recombinant circumsporozoite protein (PfCSP) was analyzed by SDS-PAGE and shown to migrate ~10 kDa higher than its predicted 30 kDa molecular weight under both reducing and non-reducing conditions. Blue-native non-denaturing PAGE and analytical size exclusion chromatography further confirmed that the rCSP monomer had a highly extended molecular structure and its size was equivalent to a ~60 kDa globular protein. Vaccination of PfCSP along with an adjuvant Montanide ISA720 induced high titer antibodies in mice and this vaccination conferred sterile protection in two strains of mice against challenge with a transgenic Plasmodium berghei parasite line that expressed the P. falciparum CSP gene. The anti-PfCSP antibodies recognized the native CSP on sporozoites by IFA and inhibited the invasion of NF54 strain sporozoites into hepatocytes. Availability of a high quality near full-length rCSP and the transgenic mouse protection model will allow us to develop novel strategies to enhance the protective efficacy of CSP based vaccines in humans.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
