Compositions and methods of vaccination against dengue virus in children and young adults

A pharmaceutical composition comprising a tetravalent vaccine formulation including all four dengue virus serotypes including a live, attenuated dengue-2 virus and dengue-2/1, dengue-2/3, and dengue-2/4 chimeras,

• wherein the dengue-2 virus is in the form of the DENV-2 16681 derived DEN-2 PDK-53 variant with a triple mutation at NS1-53, at 5′NC-57 and at NS3-250 such that the amino acid position 250 of the NS3 protein contains a valine residue, and wherein the chimeras have said DEN-2 PDK-53 genome as viral backbone and one or more structural protein genes encoding capsid, premembrane/membrane or envelope of said DEN-2 PDK-53 genome or combinations thereof replaced with one or more corresponding structural protein genes from DEN-1, DEN-3 or DEN-4,
• wherein the dengue virus serotype 2 has at least one further mutation selected from the group consisting of:
• an amino acid at position 52 of the prM protein contains a glutamic acid residue;
• and
• an amino acid at position 412 of the NS5 protein contains a valine residue;
• and
• wherein a concentration of the live attenuated dengue-2 virus serotype in the composition is at least one half a log plaque forming unit (PFU) lower than the log PFU of more than one of other dengue virus serotypes;
• or
• a concentration of the dengue-2/3 chimera is at least one-half a log greater in terms of PFUs than the dengue-2 virus serotype in the tetravalent vaccine formulation.

A method for treating healthy subjects from 1 to less than 18 years against dengue virus infection, comprising:

• obtaining or preparing a tetravalent dengue virus vaccine composition ready for administration to a patient, wherein the composition includes a dengue virus vaccine formulation including all four dengue virus serotypes, including
• a dengue-2 virus serotype comprising a live, attenuated dengue-2 virus, a dengue-2/1 chimera,
• a dengue-2/3 chimera, and
• a dengue-2/4 chimera,
• and administering the composition subcutaneously to one or more of the healthy subjects,
• wherein administering the composition consists of
• administering a first dose of the composition on day 0,
• and administering a second dose of a same composition against dengue virus about 3 months after the first administration,
• wherein the dengue-2 virus serotype is in the form of the DENV-2 16681 derived DEN-2 PDK-53 variant, with a triple mutation at NS1-53, at 5′NC-57 and at NS3-250 such that an amino acid position 250 of the NS3 protein contains a valine residue,
• wherein each of the dengue-2/1, dengue-2/3, and dengue-2/4 chimeras have said DEN-2 PDK-53 genome as viral backbone and one or more structural protein genes encoding capsid, premembrane/membrane or envelope of said DEN-2 PDK-53 genome or combinations thereof replaced with one or more corresponding structural protein genes from DEN-1, DEN-3 or DEN-4, respectively,
• wherein the dengue virus serotype 2 has at least one further mutation selected from the group consisting of:
• an amino acid at position 52 of the prM protein contains a glutamic acid residue;
• and
• an amino acid at position 412 of the NS5 protein contains a valine residue;
• and
• wherein the composition induces an immune response to dengue virus in the healthy subject.