Enhancement of the expression of HCV core gene does not enhance core-specific immune response in DNA immunization: advantages of the heterologous DNA prime, protein boost immunization regimen

doi:10.1186/1479-0556-7-7

Genetic Vaccines and Therapy

Volume 7

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Alekseeva E
Sominskaya I
Skrastina D
Egorova I
Starodubova E
Kushners E
Mihailova M
Petrakova N
Bruvere R
Kozlovskaya T
Isaguliants M
Pumpens P

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Alekseeva E
Sominskaya I
Skrastina D
Egorova I
Starodubova E
Kushners E
Mihailova M
Petrakova N
Bruvere R
Kozlovskaya T
Isaguliants M
Pumpens P
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Enhancement of the expression of HCV core gene does not enhance core-specific immune response in DNA immunization: advantages of the heterologous DNA prime, protein boost immunization regimen

Ekaterina Alekseeva¹ , Irina Sominskaya¹ , Dace Skrastina¹ , Irina Egorova²^,3 , Elizaveta Starodubova² , Eriks Kushners¹ , Marija Mihailova¹ , Natalia Petrakova⁴ , Ruta Bruvere¹ , Tatyana Kozlovskaya¹ , Maria Isaguliants²^,4^,5 and Paul Pumpens¹

¹Latvian Biomedical Research and Study Centre, Ratsupites 1, Riga, LV-1067, Latvia

²Swedish Institute of Infectious Disease Control, SE-17182 Stockholm, Sweden

³Pasteur Institute, 197101 St Petersburg, Russia

⁴Microbiology and Tumorbiology Center, Karolinska Institutet, 17177 Stockholm, Sweden

⁵D.I. Ivanovsky Institute of Virology, 123098 Moscow, Russia

author email corresponding author email

Genetic Vaccines and Therapy 2009, 7:7doi:10.1186/1479-0556-7-7


Published:	8 June 2009

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Additional file 1:

Establishment of calibration curves for quantification of core expression in vitro. Recombinant core aa 1–173 in serial dilutions in the range of 10 to 25 ng (A), 10 to 100 ng (B), or 12.5 to 100 ng (C) was loaded on 15% SDS-PAAG and resolved by gel electrophoresis together with the study samples. Proteins were transferred to PVDF membrane and subjected to Western blotting with core-specific rabbit antibodies, and secondary anti-rabbit HRP-conjugated antibodies (DAKOPatts). Signals were registered using X-ray films and ECL detection system, images were saved, scanned, and signal of individual band corresponding to core was quantified by Image J http://rsb.info.nih.gov webcite, and calibration curves were built (D).

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Additional file 2:

Summary on core-specific immune responses in BALB/c and C57BL/6 mice. Summarized data of immunization experiments performed in BALB/c and C57BL6 mice. The empty vector immunized group and the control group are composed of a mixture of BALB/c (n = 7) and C57BL6 (n = 12) mice. All the other groups had been described in Figures 4 to 6.

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