Experimental infection and protection against ... - TI Pharma

Humoral immune responses to a single allele PfAMA1 vaccine in healthy malarianaïve
adults.
*----30---*----40---* ---50 --*----60---*----70---*----80---*----90---*----100--*----110--*----
120--
FVO ∆ Glyc
QNYWEHPYQKSDVYHPINEHREHPKEYEYPLHQEHTYQQEDSGEDENTLQHAYPIDHEGAEPAPQEQNLFSSIEIVERSNYMGNPWTEYMAKYDIEEVHG
3D7 ∆.Glyc ---------N----R----------------------------------------------------------------------------------
---
HB3 ∆ Glyc ---------N----R---------------------------------------------------------------------------------
K---
CAMP ∆ Glyc ---------N—N---------------Q---------------------------------------------------------------------
---
*----130--*----140--*----150--*----160--*----170--*----180--*----190--*----200--*----210--*----
220--
FVO ∆ Glyc
SGIRVDLGEDAEVAGTQYRLPSGKCPVFGKGIIIENSKTTFLKPVATGNQDLKDGGFAFPPTNPLISPMTLNGMRDFYKNNEYVKNLDELTLCSRHAGNM
3D7 ∆ Glyc ------------------------------------------T-------Y-----------E—-M-----DE—-H---D-K---------------
---
HB3 ∆ Glyc ------------------------------------------T----E--------------E--------DQ—-HL—-D-----------------
---
CAMP ∆ Glyc --------------------------------------------------------------E----------------------------------
---
*----230--*----240--*----250--*----260--*----270--*----280--*----290--*----300--*----310--*----
320--
FVO ∆ Glyc
NPDNDKNSNYKYPAVYDYNDKKCHILTIAAQENNGPRYCNKDQSKRNSMFCFRPAKDKLFENYVYLSKNVVDNWEEVCPRKNLENAKFGLWVDGNCEDIP
3D7 ∆ Glyc I----------------DK-----------------------E--------------IS-Q--------------K-------Q-------------
---
HB3 ∆ Glyc ------------------E-----------------------E------------------------------------------------------
---
CAMP ∆ Glyc ---K-E-----------DK-----------------------E---------------S-Q--------------K---------------------
---
*----330--*----340--*----350--*----360--*----370--*----380--*----390--*----400--*----410--*----
420--
FVO ∆ Glyc
HVNEFSANDLFECNKLVFELSASDQPKQYEQHLTDYEKIKEGFKNKNADMIKSAFLPTGAFKADRYKSHGKGYNWGNYNRETQKCEIFNVKPTCLINDKS
3D7 ∆ Glyc -----P-I-----------------------------------------------------------------------T-----------------
---
HB3 ∆ Glyc --------------------------------------------------------------------R----------T-----------------
--
CAMP ∆ Glyc --------------------------------------------------------------------------------K-H--------------
---
*----430--*----440--*----450--*----460--*----470--*----480--*----490--*----500--*----510--*----
520--
FVO ∆ Glyc
YIATTALSHPIEVEHNFPCSLYKDEIKKEIERESKRIKLNDNDDEGNKKIIAPRIFISDDKDSLKCPCDPEMVSQSTCRFFVCKCVERRAEVTSNNEVVV
3D7 ∆ Glyc --------------N-----------M----------------------------------------------------------------------
---
HB3 ∆ Glyc ----------N---N--------------------------------------------------------I------N--------K---------
---
CAMP ∆ GlyC --------------N--------N—-M----------------------------------------------------------------------
----
*----530--*----540--*
FVO ∆ Glyc KEEYKEDYADIPEHKPTYDNM
3D7 ∆ Glyc -------------------K-
HB3 ∆ Glyc ---------------------
CAMP ∆ Glyc ---------------------
Figure 1: Alignment of AMA1 protein sequences amino acids 25 through 545 used for Elisa
measurements. Potential N-glycosylation sited were changed (six for FVO, 3D7 and CAMP
and five for HB3) to avoid unwanted N-glycosylation: N 162 K (For FVO, 3D7 and CAMP.
HB3 AMA1 has K at position 162). T 288 V, S 273 D, N 422 D, S 423 K and N 499 Q.
(Alhydrogel, Montanide ISA 720 or AS02) as previously described [19].
Alhydrogel and Montanide ISA 720 vaccines were prepared with lot B PfAMA1
and AS02 vaccines were prepared with lot C PfAMA1. Formulated vaccines were
kept at 4°C for a maximum of six hours until administration. Vaccine
formulations were tested for stability as previously described [20] and all
fulfilled the pre-set specifications. Immunisations were given intramuscularly in
the deltoid region of alternate arms at days 0, 28 and 56. Blood was collected in
VacutainerTM CPT tubes (Becton and Dickinson), plasma collected after
centrifugation and stored at -20°C.
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