TY - JOUR

T1 - Efficiency of HPV 16 L1/E7 DNA immunization

T2 - Influence of cellular localization and capsid assembly

AU - Kuck, Dirk

AU - Leder, Christoph

AU - Kern, Andrea

AU - Müller, Martin

AU - Piuko, Konrad

AU - Gissmann, Lutz

AU - Kleinschmidt, Jürgen A.

N1 - Funding Information: We thank Nico Michel and Corinna Klein for help with the ELISPOT assay, Petra Galmbacher for the L1-VLP production and Martin Friedel from the DKFZ animal facility. We are grateful to Harald zur Hausen for his continuous support. J.A.K. and M.M. were supported by grant 10–1912 Kl 1 of the Deutsche Krebshilfe.

PY - 2006/4/5

Y1 - 2006/4/5

N2 - Infections by human papillomaviruses (HPV) are the major cause of uterine cancer in women worldwide. Aiming to develop a combined prophylactic and therapeutic vaccine we have previously demonstrated immunogenicity of chimeric virus-like particles consisting of a C-terminally truncated HPV 16 L1 capsid protein fused to an E7 portion. Here we show that genetic vaccination with a corresponding DNA was inefficient in the induction of a L1-specific prophylactic immune response. DNA immunization with C-terminally truncated HPV 16 L1 genes of different lengths revealed that only short deletions (L1(1-498)) were tolerated for eliciting a humoral immune response against viral capsids. This correlates with the observation that the C-terminal sequences are critical for nuclear localization, capsomere and capsid assembly. However, only the ability of L1 protein to form capsomeres or capsids showed a direct influence on the outcome of the immune response. C-terminal insertion of 60 amino acids of E7 was tolerated in fusion constructs, whereas insertion of full-length E7(1-98) or shuffled E7 (149 aa) completely abolished the humoral immune response. The L1(1-498)/E7(1-60) fusion construct not only induced L1-specific antibodies but also L1- and E7-specific CTL responses after DNA vaccination.

AB - Infections by human papillomaviruses (HPV) are the major cause of uterine cancer in women worldwide. Aiming to develop a combined prophylactic and therapeutic vaccine we have previously demonstrated immunogenicity of chimeric virus-like particles consisting of a C-terminally truncated HPV 16 L1 capsid protein fused to an E7 portion. Here we show that genetic vaccination with a corresponding DNA was inefficient in the induction of a L1-specific prophylactic immune response. DNA immunization with C-terminally truncated HPV 16 L1 genes of different lengths revealed that only short deletions (L1(1-498)) were tolerated for eliciting a humoral immune response against viral capsids. This correlates with the observation that the C-terminal sequences are critical for nuclear localization, capsomere and capsid assembly. However, only the ability of L1 protein to form capsomeres or capsids showed a direct influence on the outcome of the immune response. C-terminal insertion of 60 amino acids of E7 was tolerated in fusion constructs, whereas insertion of full-length E7(1-98) or shuffled E7 (149 aa) completely abolished the humoral immune response. The L1(1-498)/E7(1-60) fusion construct not only induced L1-specific antibodies but also L1- and E7-specific CTL responses after DNA vaccination.

KW - Biology

KW - DNA immunization

KW - Papillomavirus

UR - http://www.scopus.com/inward/record.url?scp=33644880219&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/e672d426-7b3b-3d2e-acd8-e960a50761f0/

U2 - 10.1016/j.vaccine.2005.12.023

DO - 10.1016/j.vaccine.2005.12.023

M3 - Journal articles

C2 - 16414157

VL - 24

SP - 2952

EP - 2965

JO - Vaccine

JF - Vaccine

SN - 0264-410X

IS - 15

ER -