Table 1: Summary of mouse immunogenicity studies with different EV71 vaccine candidates.

EV71 vaccine candidates Adjuvant formulated Total IgG titers Western blot Virus neutralization titer
VP1-43VP2-28rVP1rVP2rVP3EV71 (B4 E59)CVA16 (N5079)

VP1-43Alum 1/3200 ++<1/8<1/8
VP1-43CFA/IFA 1/6400+++ 1/32<1/8
VP2-28Alum 1/200 <1/8 <1/8
VP2-28CFA/IFA 1/1600 +<1/8<1/8
VP1-43/VP2-28 CFA/IFA 1/6400 1/800+++ + 1/32 <1/8

rVP1Alum 1/6400 +++ <1/8 <1/8
rVP1CFA/IFA 1/12800 +++ 1/128<1/8
rVP2Alum <1/100 +++ <1/8 <1/8
rVP2 CFA/IFA 1/200 +++ 1/8 <1/8
rVP3 Alum ++<1/8 <1/8
rVP3 CFA/IFA +++ 1/8 <1/8

EV71-VLPPBS 1/1600 <1/100 ++ ++++ 1/64 <1/8
EV71-VLP Alum1/6400 <1/100 ++ ++ ++ 1/128 <1/8
EV71-VLP CFA/IFA1/12800 <1/100 +++ +++ ++ 1/160 <1/8

2 μg of inactivated
 EV71 virion PBS 1/6400 <1/100 ++++++ ++ 1/64 <1/8
 EV71 virion Alum 1/12800 <1/100 +++ +++ ++ 1/640 <1/8

5 μg of inactivated
 EV71 virion Alum 1/12800 <1/100 +++ +++ +++1/2560 <1/8

Three to six mice per group were immunized 3 times with EV71 vaccine candidates formulated with alum or CFA/IFA. Fifty micrograms of synthetic peptides, 20 μg of recombinant EV71 viral structural proteins, and 5 μg of recombinant virus-like particles (EV71-VLP) were used in the mouse immunogenicity studies. The protocols for IgG titer determination, western blot analysis, and virus neutralization assay are described in Section 2.