Correlation between log(c-day 11 parasitemia) and serologic response in animals vaccinated with FVO strain Ag.

<p>Animals were vaccinated with either <i>E. coli</i> or baculovirus expressed MSP1₄₂ in combination with FA, ISA-720 or AS02A. Growth/Invasion inhibition is against FVO (Panel A) or 3D7 (Panel B) strain targets. ELISA was performed with recombinant FVO (Panel C) or 3D7 (Panel D) strain MSP1₄₂ expressed in baculovirus. Correlations were determined with Pearson's test after testing data distributions for normality.</p

C-day 11 parasitemias for regimens 3, 7, 8, and 9 grouped by adjuvant and antigen.

<p>Open circles, individual responses; filled circles, means; bar is the 95% confidence interval for the mean. Panel A compares grouped log(c-day 11 parasitemia) levels. Panel B shows the estimated marginal means from the GLM used to analyze the responses.</p

Results from ROC graphs for Predicting noTx by Using GIA or ELISA with Ag Expressed in Baculovirus

<p>Responses evaluated were from the twenty-seven animals in vaccine regimens 3, 6, 7, 8 and 9, vaccinated with any FVO Ag in combination with any adjuvant (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002830#pone-0002830-t001" target="_blank">Table 1</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002830#pone-0002830-t002" target="_blank">2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002830#pone-0002830-t003" target="_blank">3</a>). Responses larger than the threshold value predict noTx outcome. %Inhibition is <i>in vitro</i> growth/invasion inhibition against FVO or 3D7 strain <i>P. falciparum.</i> ELISA Ab units were determined with 3D7 or FVO strain MSP1₄₂ expressed in baculovirus.</p

Trial Design and Results for Adjuvant AS02A.

<p><i>Aotus</i> monkeys vaccinated antigens in combination with AS02A. Groups of six animals were vaccinated. Day 11 is c-day 11 parasitemia. Tx is treatment outcome: noTx, no treatment required; TxA treatment for anemia; TxP, treatment for uncontrolled parasitemia. GIA is <i>in vitro</i> growth/invasion inhibition against FVO or 3D7 strain <i>P. falciparum.</i> ELISA Ab units were determined with 3D7 or FVO strain MSP1₄₂ expressed in baculovirus. ELISA Ab titers were determined with 3D7 or FVO strain MSP1₄₂, p19, EGF1 or EGF2 expressed in <i>E. coli</i>, and assays were run only using Ag homologous to the vaccine strain.</p><p>n/d is not done. ^*, no sample available.</p

Trial Design and Results for Adjuvant CFA/FA

<p><i>Aotus</i> monkeys vaccinated with Ag in combination with FA. Groups of six animals were vaccinated. Day 11 is c-day 11 parasitemia. Tx is treatment outcome: noTx, no treatment required; TxA treatment for anemia; TxP, treatment for uncontrolled parasitemia. GIA is <i>in vitro</i> growth/invasion inhibition against FVO or 3D7 strain <i>P. falciparum.</i> ELISA Ab units were determined with 3D7 or FVO strain MSP1₄₂ expressed in baculovirus. ELISA Ab titers were determined with 3D7 or FVO strain MSP1₄₂, p19, EGF1 or EGF2 expressed in <i>E. coli</i>, and assays were run only using Ag homologous to the vaccine strain. Some results from regimens 1, 3 and 7 were reported previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002830#pone.0002830-Darko1" target="_blank">[24]</a>. n/d is not done. ^*, no sample available.</p

Individual value plots showing strain-specific Ab responses by ELISA for vaccine recipients.

<p>Responses are grouped by vaccine strain received and then by the Ag strain use for ELISA. Panel A includes all 3D7 strain vaccine recipients in the trial. Panel B includes all FVO strain vaccine recipients in the trial. Connect lines join each individual animal's ELISA Ab response measured with the FVO Ag to that measured with the 3D7 Ag. Both Ag were expressed in baculovirus.</p

Results from ROC graphs for Predicting Self-Cure by Using ELISA with FVO strain Ag expressed in <i>E. coli</i>

<p>Responses evaluated were from the twenty-seven animals in vaccine regimens 3, 6, 7, 8 and 9, vaccinated with any FVO Ag in combination with any adjuvant (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002830#pone-0002830-t001" target="_blank">Table 1</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002830#pone-0002830-t002" target="_blank">2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002830#pone-0002830-t003" target="_blank">3</a>). ELISA Ab titers were determined with FVO strain MSP1₄₂, p19, EGF1 or EGF2 expressed in <i>E. coli</i>, which are homologous to the vaccine strain.</p

ELISA Ab responses for regimens 3, 7, 8, and 9 grouped by adjuvant and antigen.

<p>Capture Ags were expressed in baculovirus. Open circles, individual responses; filled circles, means; bar is the 95% confidence interval for the mean. Panel A compares grouped ELISA Ab unit levels. Panel B shows the estimated marginal means from the GLM used to analyze the responses.</p

Cumulative Logit Models used to Evaluate the Relationship between Treatment Outcome and Adjuvant and Antigen

<p>Cumulative Logit Models used to Evaluate the Relationship between Treatment Outcome and Adjuvant and Antigen</p

Trial Design and Results for Adjuvant Montanide ISA-720

<p><i>Aotus</i> monkeys vaccinated with one of three MSP1₄₂ antigens in combination with ISA-720. Groups of six animals were vaccinated. Day 11 is c-day 11 parasitemia. Tx is treatment outcome: noTx, no treatment required; TxA treatment for anemia; TxP, treatment for uncontrolled parasitemia. GIA is <i>in vitro</i> growth/invasion inhibition against FVO or 3D7 strain <i>P. falciparum.</i> ELISA Ab units were determined with 3D7 or FVO strain MSP1₄₂ expressed in baculovirus. ELISA Ab titers were determined with 3D7 or FVO strain MSP1₄₂, p19, EGF1 or EGF2 expressed in <i>E. coli</i>, and assays were run only using Ag homologous to the vaccine strain.</p