{"id":329,"date":"2014-05-28T15:13:18","date_gmt":"2014-05-28T15:13:18","guid":{"rendered":"http:\/\/www.virologyhighlights.com\/?p=329"},"modified":"2018-05-25T08:27:24","modified_gmt":"2018-05-25T08:27:24","slug":"cell-culture-systems-for-hepatitis-research","status":"publish","type":"post","link":"https:\/\/www.elsevierblogs.com\/virology\/cell-culture-systems-for-hepatitis-research\/","title":{"rendered":"Cell culture systems for hepatitis research"},"content":{"rendered":"

Production and Characterization of High-Titer Serum-Free Cell Culture Grown Hepatitis C Virus Particles of Genotype 1-6<\/b><\/p>\n

Read the full article for free on ScienceDirect.<\/a><\/p>\n

There is a need for a vaccine against hepatitis C virus (HCV), a major human pathogen. HCV infects >150 million people worldwide, and 3\u20104 million new infections occur each year. Recent development of HCV cell culture systems now enables production of cell culture derived HCV particles (HCVcc), which could serve as antigens for a whole\u2010virus inactivated vaccine. Challenges associated with generation of such a vaccine include production of sufficient amounts of HCVcc avoiding the use of substances of animal origin such as bovine serum, routinely used during cell culture. Furthermore, effective downstream purification is made difficult by the heterogeneous density profile of HCVcc, suggested to result from differential association with lipoproteins.<\/p>\n

We aimed at addressing these challenges by producing HCVcc in hepatoma cells cultured in a commercially available growth medium without bovine serum. We succeeded in producing serum\u2010free HCV (sf\u2010HCVcc) of epidemiologically important HCV genotypes 1\u20106. The sf\u2010HCVcc had higher infectivity titers than HCVcc. Genotype 1\u20106 sf\u2010HCVcc showed a homogeneous density profile, in contrast to their highly heterogeneous HCVcc counterparts. However, they displayed similar biological properties regarding apolipoprotein E association, entry routes and sensitivity to neutralizing patient serum and human monoclonal antibodies.<\/p>\n

This methodology will facilitate future vaccine studies, since it allows production of high\u2010titer sf\u2010HCVcc. Furthermore, we believe that the homogeneous density profile of sf\u2010HCVcc particles will simplify effective density\u2010based purification. Finally, it is of great importance that sf\u2010HCVcc and HCVcc showed similar susceptibility to neutralizing antibodies, underlining the relevance of sf\u2010HCVcc as potential future vaccine antigens.<\/p>\n

\"140513<\/a>
(A) HCV genotype 1-6 infected Huh7.5 hepatoma cells produce either cell culture derived HCV (HCVcc) when cultured with growth medium + 10% fetal bovine serum (FBS) (left), or serum-free cell culture derived HCV (sf-HCVcc) when cultured with serum-free growth medium (right). Both HCVcc and sf-HCVcc are believed to be associated with host lipoproteins (yellow spheres). Genotype 1-6 sf-HCV show 0.6-2.1 log10 Focus Forming Units (FFU)\/mL higher infectivity titers than HCVcc. (B) Iodixanol gradient ultracentrifugation of HCVcc and sf-HCVcc reveals that sf-HCVcc have a homogeneous single peak density profile, whereas HCVcc have a heterogeneous density profile. Density profiles of genotype 5a HCVcc (dark blue) and sf-HCVcc (light blue) are shown as relative recovery per fraction (%) plotted against the density determined for each fraction. (C) HCVcc and sf-HCVcc show similar sensitivity to neutralizing antibodies. Neutralization of genotype 5a HCVcc (dark blue) and sf-HCVcc (light blue) by chronic phase serum from a genotype 1 infected patient (H06; a kind gift from Harvey Alter, NIH) is shown. The % neutralization is plotted against the serum dilutions (log10). Sigmoidal dose-response curves were fitted [Y = Bottom + (Top – Bottom)\/(1 + 10(Log10EC50 \u2013 X)HillSlope)]. \u201cBottom\u201d was constrained to \u201c0\u201d and \u201cTop\u201d was constrained to \u201c100\u201d.<\/figcaption><\/figure>\n

Introducing the author<\/b><\/p>\n

\"Christian<\/a>
\nChristian K. Mathiesen
\nDepartment of Infectious Diseases and Clinical Research Centre, Hvidovre Hospital, Denmark<\/p>\n

About the research<\/b><\/p>\n

Production and Characterization of High-Titer Serum-Free Cell Culture Grown Hepatitis C Virus Particles of Genotype 1-6<\/a>
\n<\/b>Virology<\/i>, Volume 458-459, June 2013, Pages 190-208
\nChristian K. Mathiesen, Tanja B. Jensen, Jannick Prentoe, Henrik Krarup, Alfredo Nicosia, Mansun Law, Jens Bukh, Judith M. Gottwein<\/p>\n

Read the full article for free on ScienceDirect.<\/a><\/p>\n","protected":false},"excerpt":{"rendered":"

Production and Characterization of High-Titer Serum-Free Cell Culture Grown Hepatitis C Virus Particles of Genotype 1-6 Read the full article for free on ScienceDirect. There is a need for a vaccine against hepatitis C virus (HCV), a major human pathogen. HCV infects >150 million people worldwide, and 3\u20104 million new infections occur each year. Recent Read More…<\/a><\/p>\n","protected":false},"author":1,"featured_media":332,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[5,629],"tags":[197,56,196,100],"class_list":["post-329","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-highlighted-article","category-virus-replication","tag-cell-culture","tag-hcv","tag-hcvcc","tag-hepatitis-c-virus"],"_links":{"self":[{"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/posts\/329","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/comments?post=329"}],"version-history":[{"count":1,"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/posts\/329\/revisions"}],"predecessor-version":[{"id":333,"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/posts\/329\/revisions\/333"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/media\/332"}],"wp:attachment":[{"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/media?parent=329"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/categories?post=329"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.elsevierblogs.com\/virology\/wp-json\/wp\/v2\/tags?post=329"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}