title: Molecular characterization of the 17D-204 yellow fever vaccine.
abstract: BACKGROUND
  The worldwide use of yellow fever (YF) live attenuated vaccines came recently under close scrutiny as rare but serious adverse events have been reported. The population identified at major risk for these safety issues were extreme ages and immunocompromised subjects. Study NCT01426243 conducted by the French National Agency for AIDS research is an ongoing interventional study to evaluate the safety of the vaccine and the specific immune responses in HIV-infected patients following 17D-204 vaccination. As a preliminary study, we characterized the molecular diversity from E gene of the single 17D-204 vaccine batch used in this clinical study.
  
  
  METHODS
  Eight vials of lyophilized 17D-204 vaccine (Stamaril, Sanofi-Pasteur, Lyon, France) of the E5499 batch were reconstituted for viral quantification, cloning and sequencing of C/prM/E region.
  
  
  RESULTS
  The average rate of virions per vial was 8.68 ± 0.07 log₁₀ genome equivalents with a low coefficient of variation (0.81%). 246 sequences of the C/prM/E region (29-33 per vials) were generated and analyzed for the eight vials, 25 (10%) being defective and excluded from analyses. 95% of sequences had at least one nucleotide mutation. The mutations were observed on 662 variant sites distributed through all over the 1995 nucleotides sequence and were mainly non-synonymous (66%). Genome variability between vaccine vials was highly homogeneous with a nucleotide distance ranging from 0.29% to 0.41%. Average p-distances observed for each vial were also homogeneous, ranging from 0.15% to 0.31%.
  
  
  CONCLUSIONS
  This study showed a homogenous YF virus RNA quantity in vaccine vials within a single lot and a low clonal diversity inter and intra vaccine vials. These results are consistent with a recent study showing that the main mechanism of attenuation resulted in the loss of diversity in the YF virus quasi-species.
authors: Univ Paris Diderot, Pres Sorbone Paris Cité, Microbiology Laboratory, Hôpital Universitaire Saint-Louis, 75475 Paris 10, France. Electronic address: maud.salmona@aphp.fr.
authors: Maud
authors: M
authors: Salmona
authors: Univ Paris Diderot, Pres Sorbone Paris Cité, Infectious Diseases Department, Hôpital Universitaire Saint-Louis, 75475 Paris 10, France.
authors: Sandrine
authors: S
authors: Gazaignes
authors: Univ Paris Diderot, Pres Sorbone Paris Cité, Microbiology Laboratory, Hôpital Universitaire Saint-Louis, 75475 Paris 10, France.
authors: Severine
authors: S
authors: Mercier-Delarue
authors: Agence Nationale de la Sécurité du Médicament, ANSM site de Lyon 321, 69007 Lyon, France.
authors: Fabienne
authors: F
authors: Garnier
authors: Agence Nationale de la Sécurité du Médicament, ANSM site de Lyon 321, 69007 Lyon, France.
authors: Jehanara
authors: J
authors: Korimbocus
authors: Univ Paris Diderot, Pres Sorbone Paris Cité, Infectious Diseases Department, Hôpital Universitaire Saint-Louis, 75475 Paris 10, France.
authors: Nathalie
authors: N
authors: Colin de Verdière
authors: Univ Paris Diderot, Pres Sorbone Paris Cité, Microbiology Laboratory, Hôpital Universitaire Saint-Louis, 75475 Paris 10, France.
authors: Jerome
authors: J
authors: LeGoff
authors: Service d'Immuno-Virologie, Institute of Emerging Diseases and Innovative Therapies, Commissariat à l'Energie Atomique, 92260 Fontenay aux Roses, France.
authors: Pierre
authors: P
authors: Roques
authors: Univ Paris Diderot, Pres Sorbone Paris Cité, Microbiology Laboratory, Hôpital Universitaire Saint-Louis, 75475 Paris 10, France.
authors: François
authors: F
authors: Simon
mesh_terms: D003001:Cloning, Molecular; D016000:Cluster Analysis; D060085:Coinfection; D005602:France; D014644:Genetic Variation; D015658:HIV Infections / Q000276:immunology; D006801:Humans; D017422:Sequence Analysis, DNA; D014613:Vaccines, Attenuated; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology; D015004:Yellow Fever / Q000517:prevention & control*; D022341:Yellow Fever Vaccine / Q000009:adverse effects / Q000235:genetics* / Q000276:immunology*; D015005:Yellow fever virus / Q000145:classification / Q000235:genetics / Q000276:immunology*
chemical_list: D014613:Vaccines, Attenuated; D014764:Viral Proteins; D022341:Yellow Fever Vaccine
keywords: Genetic diversity; Vaccine; Yellow fever