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. 2020 Jun 19;147(12):1255–1262. doi: 10.1017/S003118202000102X

The use of proteomics for the identification of promising vaccine and diagnostic biomarkers in Plasmodium falciparum

Reza Mansouri 1, Mohammad Ali-Hassanzadeh 2, Reza Shafiei 3, Amir Savardashtaki 4, Mohammadreza Karimazar 5, Enayat Anvari 6, Paul Nguewa 7,✉, Sajad Rashidi 5,✉
PMCID: PMC10317765  PMID: 32618524

Abstract

Plasmodium falciparum is the main cause of severe malaria in humans that can lead to death. There is growing evidence of drug-resistance in P. falciparum treatment, and the design of effective vaccines remains an ongoing strategy to control the disease. On the other hand, the recognition of specific diagnostic markers for P. falciparum can accelerate the diagnosis of this parasite in the early stages of infection. Therefore, the identification of novel antigenic proteins especially by proteomic tools is urgent for vaccination and diagnosis of P. falciparum. The proteome diversity of the life cycle stages of P. falciparum, the altered proteome of P. falciparum-infected human sera and altered proteins in P. falciparum-infected erythrocytes could be proposed as appropriate proteins for the aforementioned aims. Accordingly, this review highlights and proposes different proteins identified using proteomic approaches as promising markers in the diagnosis and vaccination of P. falciparum. It seems that most of the candidates identified in this study were able to elicit immune responses in the P. falciparum-infected hosts and they also played major roles in the life cycle, pathogenicity and key pathways of this parasite.

Key words: Diagnostic marker, malaria, Plasmodium falciparum, proteomics, vaccine target

Introduction

Protozoan parasites from the Plasmodium genus are responsible for malaria, an infectious disease in humans and primates. Plasmodium falciparum is the main cause of severe malaria in humans that can lead to death (Castelli et al., 2010; Deress and Girma, 2019). Although chemotherapy is a current option to fight against malaria, there is growing evidence of drug-resistance in malaria treatment (Antony and Parija, 2016; Menard and Dondorp, 2017). Due to the absence of an effective vaccine against malaria, the identification of new vaccine targets is urgent for the control of malaria infection. On the other hand, the recognition of specific diagnostic markers for P. falciparum can accelerate the diagnosis of the parasite in the early stages of the infection. Therefore, the discovery of novel diagnostic markers with a high sensitivity and specificity especially by using proteomic techniques might lead to more appropriate management and might be helpful in better choosing prophylaxis for P. falciparum infection (Mathema and Na-Bangchang, 2015).

Proteomic techniques allow shedding some light on the functions, structures, and post-translation modifications of proteins from different cells including microorganisms. In addition, proteomic data also provide valuable information regarding altered proteins in response to specific cell signals, functional proteins involved in critical pathways, mechanisms of pathogenicity, and immunobiological processes (Siqueira-Batista et al., 2012; Gebretsadik and Menon, 2016; Aslam et al., 2017; Swearingen and Lindner, 2018).

The collection of data from the pathogen's proteome and the analysis of information from the host's immune responses induced after infection are considered important steps in proteomic studies which are useful in the detection and design of vaccine targets (Galassie and Link, 2015). The identification of the pathogen's proteome using two-dimensional gel electrophoresis technique followed by Western blotting assay is helpful for the achievement of immunoreactive proteins which are able to stimulate the host's immune responses (Gomase et al., 2010). As aforementioned, proteomic tools also provide a powerful means that can be used for the discovery of biomarkers for diseases. In recent years, research on the proteome profile of tissues, cells and sera from patients in comparison with healthy individuals, and also the expression of specific proteins in different microorganisms have been used for biomarker discovery in different diseases such as cancers (Gelhaus et al., 2005; Koncarevic et al., 2007; Hudler et al., 2014; Chan et al., 2016; Alharbi, 2020).

Interestingly, the genome sequencing of P. falciparum has revealed that the parasite harbours more than 5000 genes and has provided information for the discovery of novel vaccine antigens and diagnostic markers for malaria (Gardner et al., 2002; Lucchi et al., 2013; Davies et al., 2015). Although genomic investigations have facilitated the understanding of the host's immune responses, further research needs to be performed. In this context, proteomics may allow potential new diagnostic and vaccine antigens to be identified with a higher sensitivity. Accordingly, in parallel with genomic data, proteomic techniques have been proposed as useful and reliable tools for vaccine design and the discovery of diagnostic biomarkers in malaria parasites (Doolan et al., 2003; Mathema and Na-Bangchang, 2015; Sam-Yellowe, 2015).

The proteome diversity of the life cycle stages of P. falciparum, the altered proteome of P. falciparum-infected human sera and altered proteins in P. falciparum-infected erythrocytes might be related to the pathogenicity of P. falciparum and the induction of infected-host immune responses. Such proteins might provide potential and promising antigens for the design of vaccine and diagnostic targets for P. falciparum. Therefore, the current review highlights and reports on the proteins that have been identified using proteomic approaches.

Possible vaccine and diagnostic markers for P. falciparum infection (P. falciparum-life cycle stages)

The morphology of Plasmodium spp. alters across the different stages of the parasite life cycle. The specific proteome of each stage in different physiological conditions might lead to the induction of diverse immune responses in the host's immune system. On the other hand, the study of the expression of specific proteins in the parasite's life cycle stages is an important strategy for the diagnosis of infection. Therefore, the use of proteomics may be helpful for the discovery of possible vaccine and diagnostic targets for each disease stage (Florens et al., 2002; Gelhaus et al., 2005; Koncarevic et al., 2007).

Sporozoite

Sporozoite surface antigens are considered as potential stimulants of humoral immune responses and can be suggested as promising candidates for the design of malaria vaccines. The validation of such antigens has shown that the components of the inner membrane complex of live sporozoites are accessible to antibodies and stimulate the host's immune system (Swearingen et al., 2016). Accordingly, two glycosylated Plasmodium surface proteins including thrombospondin-related adhesion protein (TRAP) and circumsporozoite protein (CSP) have been suggested as potential vaccine targets against P. falciparum (Swearingen et al., 2016). Such targets have been investigated in malaria vaccination clinical trials (Moreno and Joyner, 2015). TRAP vaccine was able to induce strong T-cell responses in human volunteers using a heterologous prime-boost immunization regimen (Kimani et al., 2014). The results of phase 1 clinical trials for the CSP vaccine revealed that poor antibody responses, moderate induction of IFN-γ and TNF-α secreting CD8T cells were related to vaccine efficacy (Ouédraogo et al., 2013). Such results suggested that the improvement of effective prime-boost immunization regimens might further coordinate cellular and humoral responses to achieve more effective responses. Overall, the sterile protection obtained using TRAP and CSP vaccines was 13 and 7% in immunized volunteers, respectively (Hodgson et al., 2018).

The blockage of sporozoites, as the initial stage of malaria infection, is another appropriate strategy in the development of malaria vaccines. The identification of the sporozoite proteome provides invaluable information for this aim. Several proteins including hexose transporter, CSP, thrombospondin-related sporozoite protein (TRSP), TRAP, sporozoite conserved orthologous transcript, sugar transporter, apical membrane antigen 1 (AMA1), gamete egress and sporozoite traversal protein (GEST) and conserved Plasmodium membrane protein have been identified as surface-exposed sporozoite proteins in P. falciparum and P. yoelii (Lindner et al., 2013). Both the immunogenicity and safety of AMA1 have been evaluated for the design of malaria vaccines in children in some previous studies (phase 1 randomized controlled trial) (Hu et al., 2008; Thera et al., 2010; Jahangiri et al., 2019). A good safety profile, very robust antibody responses and acceptable tolerability were observed (Thera et al., 2010). The administration of the AMA1-based malaria vaccine FMP2.1/AS02A was suggested for assessment in a phase 2 efficacy trial in children aged 1–6 years (Thera et al., 2010). FMP2.1/AS02A was a recombinant protein (FMP2.1) based on AMA1, formulated in the Adjuvant System AS02A. Current evidence shows that multivalent malaria subunit vaccines containing some of the aforementioned proteins in combination with CSP are able to generate extensive immune responses and strong protection against Plasmodium parasites in infected hosts (Lindner et al., 2013; Moreno and Joyner, 2015).

Schizont/merozoite

Through proteomic analyses, merozoite surface protein (MSP)-1, MSP-2, MSP-4, MSP-5, MSP-10, apical sushi protein, rhoptry-associated membrane antigen (RAMA), Pf12, Pf34, Pf38 and Pf92 have been identified as glycosylphosphatidylinositol (GPI)-anchored schizont/merozoite proteins in P. falciparum (Gilson et al., 2006). Although several studies have suggested MSPs as promising for malaria vaccines, antibody production against MSP-119 and MSP-3 showed the strongest correlation with the lower incidence of malaria and protection (Chauhan et al., 2010). Accordingly, the ICGEB malaria vaccine program developed MSP-Fu24, a fusion protein, containing the conserved regions of MSP-119 and MSP-3. This strategy showed the huge potential of MSP-based malaria vaccines. However, further investigations are required concerning their clinical development (Chauhan et al., 2010). The role of RAMA has been highlighted in the development of specific immune responses in people repeatedly exposed to the P. falciparum parasite (Topolska et al., 2004). Given the vital role of rhoptry proteins, especially Pf34, during RBC invasion, such proteins have been recently proposed as promising vaccine candidates (Arévalo-Pinzón et al., 2010). Most of the surface proteins of the extracellular forms of human Plasmodium parasites are attached to the plasma membrane through GPI-anchors. Since GPI-anchored proteins are exposed to antibodies, such proteins could well be considered as possible vaccine targets in malaria (Richie and Saul, 2002).

Blood-stages of P. falciparum efficiently induce IgG responses. However, more information is still needed to detect P. falciparum antigens which are targeted by protective and non-protective IgG antibodies (Healer et al., 2018). Using IgG-immunoproteomics, two P. falciparum proteins have been characterized: protein disulphide isomerase 8 (PDI8) and StAR-related lipid transfer (START) protein as immunoreactive proteins which are targeted by IgG antibodies (Azcárate et al., 2019). Since even low levels of such antigens are able to stimulate humoral immune responses, they therefore might be proposed as appropriate diagnostic and vaccine candidates, especially in subclinical malaria in children. Similarly, further identification of several immunodominant proteins (targeted by IgG antibodies) containing PDI, elongation factor-1 α (EF-1α), 78 kDa glucose-regulated protein homologue (GRP-78), phosphoglycerate kinase (PGK), rhoptry-associated protein 2 (RAP-2) and RAP-3 confirmed the immunogenicity of PDI in P. falciparum (Costa et al., 2013).

The variant surface antigens (VSAs) including P. falciparum erythrocyte membrane protein 1 (PfEMP1), P. falciparum-encoded repetitive interspersed families of polypeptides (RIFINs), sub-telomeric variable open reading frame (STEVOR) and surface-associated interspersed gene family (SURFIN) have been proposed as possible malaria vaccine targets (Bark et al., 2018). Besides such markers, a recent proteomic assessment identified PfJ23, parasite-infected erythrocyte (PIE) surface protein 2 (PIESP2), gametocyte exported protein 7 (GEXP07), liver stage antigen-3 (LSA-3), and PF3D7 as new vaccine targets in P. falciparum (Table 1) (Bark et al., 2018). Those previously mentioned molecules suggested as immunodominant proteins in P. falciparum have been widely applied to measure antibody responses against malaria infection in humans.

Table 1.

Possible vaccine and diagnostic biomarkers in P. falciparum-life cycle stages

Possible vaccine and diagnostic biomarkers
Sporozoite: TRAP, CSP, hexose transporter, CSP, TRSP, sporozoite conserved orthologous transcript, sugar transporter, AMA1, GEST, conserved Plasmodium membrane protein
Schizont/merozoite: MSP-1, MSP-2, MSP-4, MSP-5, MSP-10, apical sushi protein, RAMA, Pf12, Pf34, Pf38, Pf92, PDI 8, START, PfJ23, PIESP2, GEXP07, LSA3, PF3D7, HSP70-1, HSP70-x, EF-1α, PGK, GRP-78, RAP-2, RAP-3
Gametocytes: Pfs48/45, Pfg27, P47, plasma-membrane associated protein Pfs230, CCp, Pfg377, the orthologue of the OB (osmiophilic bodies) component of the rodent malaria gametocytes PbGEST, PfDPAP2, PfSUB2, an unknown 13 kDa protein
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Some of the mechanisms of malaria pathogenesis refer to proteins involved in the surface infection of RBCs. In 2017, an immunoproteomic study based on immunoreactive proteins of P. falciparum-infected RBCs identified chaperones such as heat shock protein 70-1 (HSP70-1) and HSP70-x [as components of the secretion machinery/the putative Plasmodium translocon of exported proteins (PTEX)] as P. falciparum immunodominant proteins (Cabral et al., 2017). The use of HSPs as malaria DNA vaccines or adjuvants in subunit vaccines had been applied in previous studies (Sanchez et al., 2001; Qazi et al., 2005).

Gametocytes

Gametocytogenesis is defined as the sexual differentiation of asexual precursors. It occurs during the Plasmodium blood-stage in hosts. Successful malaria transmission takes place through the availability of gametocytes in human peripheral blood during the feeding of Anopheles mosquitoes. Due to the insignificant role of gametocytes in the clinical symptoms of malaria, gametocytes are less commonly used for the design of therapeutic targets in treatment strategies against malaria. The proteome identification of Plasmodium gametocytes might further allow the design of new malaria transmission-blocking vaccines (Table 1) (Frimpong et al., 2018).

The specific proteins of male and female gametocytes of P. falciparum are different in structure and function. It seems that such proteins are related to genome replication and flagella. However, specific proteins belonging to female gametocytes were mostly involved in translation, metabolism and organellar functions. In addition, the comparison of gametocyte-specific proteins (male and female) in Plasmodium spp. revealed the important function of such proteins in the cytoskeleton, protein degradation and lipid metabolism (Miao et al., 2017). It was also shown that the male gametocyte proteins were mostly involved in the configuration of flagellated gametes, chromatin organization, DNA replication and axoneme formation. In addition, many proteins of female gametocytes were correlated with zygote configuration and important functions after fertilization; lipid, protein and energy metabolism (Lasonder et al., 2016). Furthermore, a comparative proteomic study demonstrated the high expression of antioxidant proteins in young gametocytes and the upregulation of membrane-associated proteins in mature gametocytes (Thima et al., 2017). Altogether, these data might be helpful to better understand the complex biology of malaria parasites and to make advances in the design of appropriate vaccine candidates in the future.

Malaria transmission-blocking vaccines were designed to disrupt parasite transmission between mosquito vectors and humans. It has been shown that several proteins in P. falciparum gametocytes are able to potentially activate host immunological responses (Frimpong et al., 2018). ATPase-activity altered in Plasmodium parasites from asexual schizonts to sexual gametocytes suggested the regulatory role of malarial-ATPases and the complex metabolism of malaria parasites in different stages (Ortega et al., 2018). Therefore, the characterization of specific malarial-ATPases including Pfs48/45, Pfg27 and the plasma membrane-associated protein Pfs230 with gametocytogenesis might provide valuable information regarding the inhibition of malaria transmission (Simon et al., 2016; Ortega et al., 2018). Pfs48/45 is an important factor in the malaria parasite's adherence to host cells and was interestingly suggested as a possible vaccine candidate for blocking malaria transmission (Singh et al., 2015). In addition to Pfs48/45 and Pfs230, other possible vaccine targets such as LCCL domain-containing protein (CCp) and P47 have been identified in the proteome of P. falciparum stage V gametocyte (Tao et al., 2014).

Osmiophilic bodies (OBs) are considered to be electron-dense secretory organelles in the female gametocytes of Plasmodium spp. with a possible role in gamete egress phase of the life cycle of these parasites (Blackman and Bannister, 2001; Souza, 2006). A recent study has indicated the role of several proteins related to OBs including Pfg377, the orthologue of the OB component of the rodent malaria gametocytes PbGEST, dipeptidyl aminopeptidase 2 (PfDPAP2), subtilisin 2 (PfSUB2) and an unknown 13 kDa protein in gamete egress and oocyst formation in P. falciparum. The aforementioned targets might be suggested as valuable markers in the biology of Plasmodium spp. such as interrupting malaria transmission (designing malaria transmission-blocking vaccines) (Suárez-Cortés et al., 2016).

Plasmodium falciparum-infected human sera

Alteration of the proteome of P. falciparum-infected human sera occurs during malaria pathogenicity. The identification of abnormal markers might enhance our understanding of malaria pathogenicity and boost the discovery of potential diagnostic biomarkers. Moreover, host immune responses could be evaluated by targeting such proteins for vaccine candidate design.

In 2012, the modulation of several important physiological pathways containing cytokine and chemokine signalling, acute phase response signalling, complement cascades and blood coagulation was detected in P. falciparum-infected human sera (Ray et al., 2012). That study suggested a panel of modulated proteins such as serum amyloid A, haptoglobin (HAP), apolipoprotein E, hemopexin, apolipoprotein A-I and retinol-binding protein as possible diagnostic markers in P. falciparum-infected human sera (Ray et al., 2012). The detection of serum amyloid A and HAP in patients infected with P. vivax supported the selection of these proteins as potential diagnostic biomarkers for malaria (Bahk et al., 2010).

Systemic inflammation and sequestration of P. falciparum-infected RBCs are considered as major processes in the pathophysiology of severe childhood malaria induced by P. falciparum (Ponsford et al., 2012). The proteome identification of severe and uncomplicated malaria might lead to a better understanding of the pathogenicity mechanisms and differentiation of severe and uncomplicated forms of childhood malaria. Interestingly, the expression of biomarkers related to oxidative stress was observed in malaria-infected children with anaemia. Moreover, specific markers related to platelet adhesion, muscular damage and endothelial activation were detected in infected children with cerebral malaria (CM) (Bachmann et al., 2014). Other changes were described: generalized vascular inflammation, activation of endothelium, vascular wall modulations and irregular glucose metabolism in patients with severe malaria. The release of a high level of specific muscle markers into plasma led to the induction of microvasculature lesions and severe muscle damage in patients with CM.

It is known that C-reactive protein (CRP), tumour necrosis factor (TNF), lymphocyte cytosolic protein 1 (LCP1), vascular cell adhesion molecule 1 (VCAM1), insulin-like growth factor-binding protein 1 (IGFBP1), integrin subunit α V (ITGAV), matrix metalloproteinase 2 (MMP2), calcitonin-related polypeptide α (CALCA), adenylosuccinate synthase-like 1 (ADSSL1), timeless-interacting protein (TIPIN) and myosin light chain 3 (MYL3) were upregulated proteins in the plasma proteome of patients with acute paediatric malaria (induced by P. falciparum) (Table 2). In contrast, other proteins including osteonectin (anti-adhesive SPARC), apoptotic CTSD and cell migration chemoattractant CCL5 (RANTES) were downregulated in patients in comparison with the control group (Reuterswärd et al., 2018). LCP1 and VCAM1 played a role in cell migration and cell adhesion, respectively. Both cellular functions (adhesion and migration) seemed to be associated with IGFBP1 and ITGAV (Reuterswärd et al., 2018). Exploration of the plasma/serum proteome in malaria-infected patients reveals more information concerning the pathophysiology of malaria and might lead to more effective treatment, diagnosis and vaccination against the disease. Although the diagnostic and prognostic potential of CRP, TNF and VCAM1 have been evaluated in previous malaria investigations, more studies will be needed to validate the aforementioned diagnostic biomarkers in P. falciparum (Ohnishi and Kimura, 2001; Perera et al., 2013; Sarfo et al., 2018).

Table 2.

Possible diagnostic biomarkers in P. falciparum-infected human sera

Protein biomarkers
Serum amyloid A, haptoglobin, apolipoprotein E, hemopexin, apolipoprotein A-I, retinol-binding protein, lipopolysaccharide-binding protein (LBP), Von Willebrand factor (VWF), colony-stimulating factor 1 (CSF1), neurofilament medium (NEFM), CRP, TNF, LCP1, VCAM1, IGFBP1, ITGAV, MMP2, CALCA, ADSSL1, TIPIN, MYL3, osteonectin, apoptotic CTSD, cell migration chemoattractant CCL5, CADM4, HAP, Gal3BP, titin, spectrin β chain brain 3, SGLT1, EMP 1, rifins, HSP40, AP2 domain transcription factor, an armadillo repeat protein, NLI-interacting factor-like phosphatase
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The identification of common diagnostic proteins in Plasmodium spp. using proteomic approaches might facilitate the design of diagnostic tests that are able to detect mixed malaria infections (Ray et al., 2012). Interestingly, the upregulation of CRP and adhesion molecule-4 (CADM4) was observed in pooled sera of patients infected with P. knowlesi, P. falciparum and P. vivax. However, serum levels of HAP were decreased in pooled sera (Mu et al., 2014).

The information inferred from the sera-proteome of patients infected with severe and non-severe malaria (caused by P. falciparum) has elucidated the modulation of some critical pathways such as IL-12 signalling and production in macrophages, chemokine and cytokine signalling, blood coagulation, complement cascades and protein ubiquitination pathways (Ray et al., 2015). The overexpression of muscle and cytoskeletal-related proteins containing the galectin-3-binding protein-(Gal3BP) and titin has been reported in the sera of patients suffering severe malaria. Due to the differential expression of CRP, serum amyloid A, HAP and apolipoprotein E in patients' sera, such proteins might be applied as biomarkers for the determination of the degree of severity of malaria in patients (Ray et al., 2015).

Since P. falciparum is the causative agent of a severe form of malaria (CM), and based on the similar clinical symptoms of CM in children and other non-malarial encephalopathies, proteome identification of plasma and cerebrospinal fluid (CSF) can provide specific biomarkers for differentiation and treatment of these disorders. The differential expression of spectrin β chain brain 3 (a host protein) has been reported in CSF and plasma of children infected with CM (Gitau et al., 2013). This protein was determined as a P. falciparum-binding partner with several possible functions involved in the suppression of parasite invasion, the stability of infected-RBCs, and increasing the ability of RBCs to sequester in the microvasculature. The overexpression of sodium/glucose cotransporter 1 (SGLT1), a host protein involved in metabolic stress, has been also reported in the CSF from patients with CM. Furthermore, different variants of EMP 1 and RIFINs (with functions in host cell interaction and antigenic variation) and HSP40 (involved in host cell modification) were discovered as specific proteins belonging to the parasite P. falciparum (Gitau et al., 2013).

It has been shown that a pre-erythrocytic long-lasting sterile protection is induced by immunization with sporozoites under chloroquine chemoprophylaxis (CPS) against homologous controlled human malaria infection (Roestenberg et al., 2009). Three conserved proteins with unknown function (AP2 domain transcription factor, an armadillo repeat protein and NLI-interacting factor-like phosphatase) have been detected in plasma samples of CPS-immunized individuals, as protein biomarkers related to the possible protective humoral immune responses (Obiero et al., 2019).

Other possible vaccine and diagnostic biomarkers in P. falciparum

Plasmodium falciparum-infected RBCs

The presence of Plasmodium parasites in related host cells such as erythrocytes can lead to the changes in the host cells' proteome. The expression of new proteins, the exchange of proteins between parasite and the host cell, alteration in immunobiological and physicochemical functions of Plasmodium-host cells, and also upregulation and downregulation of several specific proteins might occur during the aforementioned process (Table 3) (Maier et al., 2008; Zhang et al., 2015).

Table 3.

Other possible vaccine and diagnostic biomarkers in P. falciparum

Protein biomarkers
P. falciparum-infected RBCs: PIESP1, PIESP2, MESA, Pf332, VAR2CSA, PF14-0018, PFI1785w, PFA-0410w, PFB0115w, PFF0325c
Expressed proteins in patients with CM: 14-3-3 protein, PfLDH, enolase, SERPINA1, ORM1
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RBCs, red blood cells; CM, cerebral malaria.

PIESPs are valuable biomarkers for malaria pathogenicity and useful targets for the design of novel vaccine and drugs for the control of malaria (Deitsch and Wellems, 1996). PIESP1 and PIESP2 have been recognized as two new surface markers in P. falciparum-infected RBCs using a shotgun proteomics approach. Unlike other PIESPs such as rifin and PfEMP1, PIESP1 and PIESP2 are highly conserved in Plasmodium ssp., and therefore, might be promising candidates for vaccine design. Interestingly, Florens et al. suggested the unlikely relationship between PIESP1 and PIESP2 and the protrusions on the PIE surface (Florens et al., 2004).

Mature parasite-infected erythrocyte surface antigen (MESA) and P. falciparum antigen 332 (Pf332) have been characterized as important proteins expressed on the erythrocyte membrane of patients with CM (Bertin et al., 2016). Although the antigenic function of MESA remains unclear, MESA and Pf332 were proposed as biomarkers involved in protein trafficking such as export of VSAs. Interestingly, Pf332 is able to be exposed to the immune system. A possible association of such proteins was observed with the pathophysiology of CM (Bertin et al., 2016).

Plasmodium falciparum can lead to the induction of pregnancy-associated malaria (PAM) in pregnant women. In 2013, VAR2CSA was identified as a biomarker associated with PAM using proteomic techniques. VAR2CSA is considered a member of the PfEMP1 family. This marker and other proteins (PF14-0018, PFI1785w, PFA-0410w, PFB0115w and PFF0325c) might have a possible function in PAM-pathophysiology and might support a vaccine design against PAM (Bertin et al., 2013).

Although VAR2CSA was suggested as a vaccine target in PAM and reached the clinical development stage, it seemed that the high variability and large size of this protein might restrict its efficacy for vaccine design (Badaut et al., 2010; Hviid, 2010; Fried and Duffy, 2015). Recently, an interesting study confirmed the upregulation of the PFI1785w protein in PAM. This antigen is highly conserved in comparison with VAR2CSA and can play a major role in the pathogenesis of PAM. Therefore, the use of alternative antigens in combination with VAR2CSA might offer a new perspective on vaccine strategies against PAM (Kamaliddin et al., 2017).

Expressed proteins in patients with CM

The host responses (proteome of cells, RBCs, issues and immune responses) in patients infected with malaria might be altered due to the presence of Plasmodium parasites in the host (Kumar et al., 2018). Recently, 14-3-3 protein, L-lactate dehydrogenase (PfLDH) and enolase have been identified as P. falciparum-specific biomarkers in the brain proteome of patients with CM (Kumar et al., 2018). In 2006, LDH was described as a promising protein for rapid malaria diagnosis (Seydel et al., 2006; Hviid, 2010). Enolase is a protein localized on the merozoite cell surface and a potential protective antigen. Furthermore, the detection of anti-enolase antibodies in malaria patients and experimental immunized animal models confirmed the immunostimulant property of this protein (Pal-Bhowmick et al., 2007). The α-1-antitrypsin (SERPINA1) (anti-inflammatory molecule) and α-1-acid glycoprotein 1 (ORM1) have been reported as upregulated proteins relevant to the host immune system in patients with CM (Table 3) (Kumar et al., 2018). The upregulation of SERPINA3 in serum has been previously reported in malaria patients (Kassa et al., 2012).

Plasmodium falciparum secretions

The importance of proteome identification from protozoa secretions has been highlighted in recent years. Secretions of some parasites as potential activators of the host's immune responses seemed to be potential sources of antigens for the design of diagnostic and vaccine targets (Gour et al., 2012; Garg et al., 2018; Lin et al., 2019). However, proteomic investigations regarding secretory antigens of Plasmodium parasites are still urgently needed.

Most pathogens apply secretory molecules to prepare host cells for invasion, to acquire nutrients and to subvert the host's immune responses (Huynh et al., 2003; Cezairliyan and Ausubel, 2017; Belachew, 2018). According to a recent investigation, extracellular vesicles (EVs) in P. falciparum mediate the transfer of genetic material between Plasmodium parasites and induce sexual commitment (Sampaio et al., 2017). In P. falciparum, EVs are comprised of proteins that are found within the secretory endomembrane compartments in the apical end of merozoites and exomembrane compartments of infected-RBCs such as Maurer's clefts (Abdi et al., 2017). The major role of PfEVs in parasite–host interactions and the pathogenicity of P. falciparum support the idea that these proteins are potentially useful for the design of vaccine and diagnostic targets in malaria parasites.

Conclusion

Despite the important progress made in proteomic techniques, and because some limitations such as the huge cost of equipment and the need for highly trained technicians remain unsolved, the abovementioned methods have not yet been established in clinical practice. However, the success of such techniques in diagnostic biomarker discovery has been underlined in some diseases, especially cancers. Most of the proteins identified were able to activate immune responses in P. falciparum-infected hosts and also played major roles in the life cycle, pathogenicity and the important pathways of this parasite. The validation of novel biomarkers using laboratory techniques including protein recombinant production, enzyme-linked immunosorbent assays, Western blotting and quantitative real-time polymerase chain reactions might increase their usage in clinical fields. Some of the aforementioned proteins in this work including CSP, AMA-1, MSP-1, VAR2CSA, TRAP (vaccine targets) and HDL (diagnostic target) have been previously investigated for malaria vaccination and diagnosis in clinical trials. Although some of the proteins highlighted in this review failed to provide protection in clinical studies, further assessment of these proteins, for vaccine potentiation in combination with new adjuvants, might continue to make them promising targets for vaccination against P. falciparum in the future.

Acknowledgements

PN thanks Obra Social La Caixa (LCF/PR/PR13/11080005), Fundación Caja Navarra, Gobierno de Navarra-Salud (12/2017), Fundación Roviralta, Ubesol, Government of Navarre, Laser Ebro, Inversiones Garcilaso de la Vega and COST Actions CA18217 and CA18218 for their support. We acknowledge Prof Paul Miller (PhD) from the University of Navarra for language editing.

Financial support

This research received no specific grant from any funding agency, commercial or not-for-profit sectors.

Ethical standards

Not applicable.

Conflict of interest

None.

References

  1. Abdi A, Yu L, Goulding D, Rono MK, Bejon P, Choudhary J and Rayner J (2017) Proteomic analysis of extracellular vesicles from a Plasmodium falciparum Kenyan clinical isolate defines a core parasite secretome. Wellcome Open Research 2, 50. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alharbi RA (2020) Proteomics approach and techniques in identification of reliable biomarkers for diseases. Saudi Journal of Biological Sciences 27, 968–974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Antony HA and Parija SC (2016) Antimalarial drug resistance: an overview. Tropical Parasitology 6, 30–41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Arévalo-Pinzón G, Curtidor H, Vanegas M, Vizcaíno C, Patarroyo MA and Patarroyo ME (2010) Conserved high activity binding peptides from the Plasmodium falciparum Pf34 rhoptry protein inhibit merozoites in vitro invasion of red blood cells. Peptides 31, 1987–1994. [DOI] [PubMed] [Google Scholar]
  5. Aslam B, Basit M, Nisar MA, Khurshid M and Rasool MH (2017) Proteomics: technologies and their applications. Journal of Chromatographic Science 55, 182–196. [DOI] [PubMed] [Google Scholar]
  6. Azcárate IG, Marin-Garcia P, Abad P, Perez-Benavente S, Paz-Artal E, Reche PA, Fobil JN, Rubio JM, Diez A, Puyet A and Bautista JM (2019) Plasmodium falciparum immunodominant IgG epitopes in subclinical malaria. bioRxiv 792499. [DOI] [PMC free article] [PubMed]
  7. Bachmann J, Burté F, Pramana S, Conte I and Brown B (2014) Affinity proteomics reveals elevated muscle proteins in plasma of children with cerebral malaria. PLoS Pathogens 10, e1004038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Badaut C, Bertin G, Rustico T, Fievet N, Massougbodji A, Gaye A and Deloron P (2010) Towards the rational design of a candidate vaccine against pregnancy associated malaria: conserved sequences of the DBL6ɛ domain of VAR2CSA. PLoS ONE 5, e11276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bahk YY, Na BK, Cho SH, Kim JY, Lim KJ and Kim TS (2010) Proteomic analysis of haptoglobin and amyloid A protein levels in patients with vivax Malaria. The Korean Journal of Parasitology 48, 203–211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Bark SKN, Ahmad R, Dantzler K, Lukens AK, De Niz M, Szucs MJ, Jin X, Cotton J, Hoffmann D, Bric-Furlong E, Oomen R, Parrington M, Milner D, Neafsey DE, Carr SA, Wirth DF and Marti M (2018) Quantitative proteomic profiling reveals novel Plasmodium falciparum surface antigens and possible vaccine candidates. Molecular & Cellular Proteomics 17, 43–60. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Belachew EB (2018) Immune response and evasion mechanisms of Plasmodium falciparum parasites. Journal of Immunology Research 2018, 1–6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Bertin GI, Sabbagh A, Guillonneau F, Jafari-Guemouri S, Ezinmegnon S, Federici C, Hounkpatin B, Fievet N and Deloron P (2013) Differential protein expression profiles between Plasmodium falciparum parasites isolated from subjects presenting with pregnancy-associated malaria and uncomplicated malaria in Benin. The Journal of Infectious Diseases 208, 1987–1997. [DOI] [PubMed] [Google Scholar]
  13. Bertin GI, Sabbagh A, Argy N, Salnot V, Ezinmegnon S, Agbota G, Ladipo Y, Alao JM, Sagbo G, Guillonneau F and Deloron P (2016) Proteomic analysis of Plasmodium falciparum parasites from patients with cerebral and uncomplicated malaria. Scientific Reports 6, 26773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Blackman MJ and Bannister LH (2001) Apical organelles of Apicomplexa: biology and isolation by subcellular fractionation. Molecular and Biochemical Parasitology 117, 11–25. [DOI] [PubMed] [Google Scholar]
  15. Cabral FJ, Vianna LG, Medeiros MM, Carlos BC, Martha RD, Silva NM, Hildebrando P, da Silva L, Stabeli RG and Wunderlich G (2017) Immunoproteomics of Plasmodium falciparum-infected red blood cell membrane fractions. Memórias do Instituto Oswaldo Cruz 112, 850–856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Castelli F, Odolini S, Autino B, Foca E and Russo R (2010) Malaria prophylaxis: a comprehensive review. Pharmaceuticals 3, 3212–3239. [Google Scholar]
  17. Cezairliyan B and Ausubel FM (2017) Investment in secreted enzymes during nutrient-limited growth is utility dependent. Proceedings of the National Academy of Sciences 114, E7796–E7802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Chan PP, Wasinger VC and Leong RW (2016) Current application of proteomics in biomarker discovery for inflammatory bowel disease. World Journal of Gastrointestinal Pathophysiology 7, 27–37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Chauhan VS, Yazdani SS and Gaur D (2010) Malaria vaccine development based on merozoite surface proteins of Plasmodium falciparum. Human Vaccines 6, 757–762. [DOI] [PubMed] [Google Scholar]
  20. Costa RM, Nogueira F, de Sousa KP, Vitorino R and Silva MS (2013) Immunoproteomic analysis of Plasmodium falciparum antigens using sera from patients with clinical history of imported malaria. Malaria Journal 12, 100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Davies DH, Duffy P, Bodmer J-L, Felgner PL and Doolan DL (2015) Large screen approaches to identify novel malaria vaccine candidates. Vaccine 33, 7496–7505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Deitsch KW and Wellems TE (1996) Membrane modifications in erythrocytes parasitized by Plasmodium falciparum. Molecular and Biochemical Parasitology 76, 1–10. [DOI] [PubMed] [Google Scholar]
  23. Deress T and Girma M (2019) Plasmodium falciparum and Plasmodium vivax prevalence in Ethiopia: a systematic review and meta-analysis. Malaria Research and Treatment 2019, 7065064. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Doolan DL, Southwood S, Freilich DA, Sidney J, Graber NL, Shatney L, Bebris L, Florens L, Dobano C, Witney AA, Appella E, Hoffman SL, Yates JR, Carucci DJ and Sette A (2003) Identification of Plasmodium falciparum antigens by antigenic analysis of genomic and proteomic data. Proceedings of the National Academy of Sciences 100, 9952–9957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Florens L, Washburn MP, Raine JD, Anthony RM, Grainger M, Haynes JD, Moch JK, Muster N, Sacci JB, Tabb DL, Witney AA, Wolters D, Wu Y, Gardner MJ, Holder AA, Sinden RE, Yates JR and Carucci DJ (2002) A proteomic view of the Plasmodium falciparum life cycle. Nature 419, 520–526. [DOI] [PubMed] [Google Scholar]
  26. Florens L, Liu X, Wang Y, Yang S, Schwartz O, Peglar M, Carucci D, Yates JR and Wub Y (2004) Proteomics approach reveals novel proteins on the surface of malaria-infected erythrocytes. Molecular and Biochemical Parasitology 135, 1–11. [DOI] [PubMed] [Google Scholar]
  27. Fried M and Duffy PE (2015) Designing a VAR2CSA-based vaccine to prevent placental malaria. Vaccine 33, 7483–7488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Frimpong A, Kusi KA, Ofori MF and Ndifon W (2018) Novel strategies for malaria vaccine design. Frontiers in Immunology 9, 2769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Galassie AC and Link AJ (2015) Proteomic contributions to our understanding of vaccine and immune responses. PROTEOMICS – Clinical Applications 9, 972–989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Gardner MJ, Hall N, Fung E, White O, Berriman M, Hyman RW, Carlton JM, Pain A, Nelson KE, Bowman S, Paulsen IT, James K, Eisen J, Rutherford K, Salzberg SL, Craig A, Kyes S, Chan MS, Nene V, Shallom SJ, Suh B, Peterson J, Angiuoli S, Pertea M, Allen J, Selengut J, Haft D, Mather MW, Vaidya AB, Martin DMA, Fairlamb AH, Fraunholz MJ, Roos DS, Ralph SA, McFadden GI, Cummings LM, Subramanian GM, Mungall C, Venter JC, Carucci DJ, Hoffman SL, Newbold C, Davis RW, Fraser CM and Barrell B (2002) Genome sequence of the human malaria parasite Plasmodium falciparum. Nature 419, 498–511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Garg G, Singh K and Ali V (2018) Proteomic approaches unravel the intricacy of secreted proteins of Leishmania: an updated review. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics 1866, 913–923. [DOI] [PubMed] [Google Scholar]
  32. Gebretsadik G and Menon M (2016) Proteomics and its applications in diagnosis of auto immune diseases. Open Journal of Immunology 6, 14–33. [Google Scholar]
  33. Gelhaus C, Fritsch J, Krause E and Leippe M (2005) Fractionation and identification of proteins by 2-DE and MS: towards a proteomic analysis of Plasmodium falciparum. Proteomics 5, 4213–4222. [DOI] [PubMed] [Google Scholar]
  34. Gilson PR, Nebl T, Vukcevic D, Moritz RL, Sargeant T, Speed TP, Schofield L and Crabb BS (2006) Identification and stoichiometry of glycosylphosphatidylinositol-anchored membrane proteins of the human malaria parasite Plasmodium falciparum. Molecular & Cellular Proteomics 5, 1286–1299. [DOI] [PubMed] [Google Scholar]
  35. Gitau EN, Kokwaro GO, Karanja H, Newton CR and Ward SA (2013) Plasma and cerebrospinal proteomes from children with cerebral malaria differ from those of children with other encephalopathies. The Journal of Infectious Diseases 208, 1494–1503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Gomase V, Kapoor R and Ladak S (2010) Immuno-proteomics approach for synthetic vaccine development form Haemophilus influenzae. Journal of Infectious Diseases Letters 1, 39–46. [Google Scholar]
  37. Gour JK, Kumar V, Singh N, Bajpai S, Pandey HP and Singh RK (2012) Identification of Th1-responsive leishmanial excretory–secretory antigens (LESAs). Experimental Parasitology 132, 355–361. [DOI] [PubMed] [Google Scholar]
  38. Healer J, Chiu CY and Hansen DS (2018) Mechanisms of naturally acquired immunity to P. falciparum and approaches to identify merozoite antigen targets. Parasitology 145, 839–847. [DOI] [PubMed] [Google Scholar]
  39. Hodgson SH, Ewer KJ, Bliss CM, Edwards NJ, Rampling T, Anagnostou NA, de Barra E, Havelock T, Bowyer G, Poulton ID, de Cassan S, Longley R, Illingworth JJ, Douglas AD, Mange PB, Collins KA, Roberts R, Gerry S, Berrie E, Moyle S, Colloca S, Cortese R, Sinden RE, Gilbert SC, Bejon PH, Lawrie AM, Nicosia A, Faust SN and Hill AVS (2018) Evaluation of the efficacy of ChAd63-MVA vectored vaccines expressing circumsporozoite protein and ME-TRAP against controlled human malaria infection in malaria-naive individuals. The Journal of Infectious Diseases 211, 1076–1086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Hu J, Chen Z, Gu J, Wan M, Shen Q, Kieny MP, He J, Li Z, Zhang Q, Reed ZH, Zhu Y, Li W, Cao Y, Qu L, Cao Z, Wang Q, Liu H, Pan X, Huang X, Zhang D, Xue X and Pan W (2008) Safety and immunogenicity of a malaria vaccine, Plasmodium falciparum AMA-1/MSP-1 chimeric protein formulated in montanide ISA 720 in healthy adults. PLoS ONE 3, e1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Hudler P, Kocevar N and Komel R (2014) Proteomic approaches in biomarker discovery: new perspectives in cancer diagnostics. The Scientific World Journal 2014, 260348. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Huynh MH, Rabenau KE, Harper JM, Beatty WL, Sibley LD and Carruthers VB (2003) Rapid invasion of host cells by Toxoplasma requires secretion of the MIC2–M2AP adhesive protein complex. The EMBO Journal 22, 2082–2090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Hviid L (2010) The role of Plasmodium falciparum variant surface antigens in protective immunity and vaccine development. Human Vaccines 6, 84–89. [DOI] [PubMed] [Google Scholar]
  44. Jahangiri F, Jalallou N and Ebrahimi M (2019) Analysis of apical membrane antigen (AMA)-1 characteristics using bioinformatics tools in order to vaccine design against Plasmodium vivax infection. Genetics and Evolution 71, 224–231. [DOI] [PubMed] [Google Scholar]
  45. Kamaliddin C, Salnot V, Leduc M, Ezinmegnon S, Broussard C, Fievet N, Deloron P, Guillonneau F and Bertin GI (2017) PFI1785w: a highly conserved protein associated with pregnancy associated malaria. PLoS ONE 12, e0187817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Kassa FA, Shio MT, Bellemare MJ, Faye B, Ndao M and Olivier M (2012) New inflammation-related biomarkers during malaria infection. PLoS ONE 6, e26495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Kimani D, Jagne YJ, Cox M, Kimani E, Bliss CM, Gitau E, Ogwang C, Afolabi MO, Bowyer G, Collins KA, Edwards N, Hodgson SH, Duncan CJA, Spencer AJ, Knight MG, Drammeh A, Anagnostou NA, Berrie E, Moyle S, Gilbert SC, Soipei P, Okebe J, Colloca S, Cortese R, Viebig NK, Roberts R, Lawrie AM, Nicosia A, Imoukhuede EB, Bejon PH, Chilengi R, Bojang K, Flanagan KL, Hill AVS, Urban BC and Ewer KJ (2014) Translating the immunogenicity of prime-boost immunization with ChAd63 and MVA ME-TRAP from malaria naive to malaria-endemic populations. Molecular Therapy 22, 1992–2003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Koncarevic S, Bogumil R and Becker K (2007) SELDI-TOF-MS analysis of chloroquine resistant and sensitive Plasmodium falciparum strains. Proteomics 7, 711–721. [DOI] [PubMed] [Google Scholar]
  49. Kumar M, Varun CN, Dey G, Ravikumar R, Mahadevan A, Shankar SK and Prasad TK (2018) Identification of host-response in cerebral malaria patients using quantitative proteomic analysis. PROTEOMICS – Clinical Applications 12, 1600187. [DOI] [PubMed] [Google Scholar]
  50. Lasonder E, Rijpma SR, van Schaijk BC, Hoeijmakers WA, Kensche PR, Gresnigt MS, Italiaander A, Vos MW, Woestenenk R, Bousema T, Mair GR, Khan SM, Janse CJ, Bártfai R and Sauerwein RW (2016) Integrated transcriptomic and proteomic analyses of P. falciparum gametocytes: molecular insight into sex-specific processes and translational repression. Nucleic Acids Research 44, 6087–6101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Lin WC, Tsai CY, Huang JM, Wu SR, Chu LJ and Huang KY (2019) Quantitative proteomic analysis and functional characterization of Acanthamoeba castellanii exosome-like vesicles. Parasites & Vectors 12, 1–12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Lindner SE, Swearingen KE, Harupa A, Vaughan AM, Sinnis P, Moritz RL and Kappe SH (2013) Total and putative surface proteomics of malaria parasite salivary gland sporozoites. Molecular & Cellular Proteomics 12, 1127–1143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Lucchi N, Oberstaller J, Kissinger J and Udhayakumar V (2013) Malaria diagnostics and surveillance in the post-genomic era. Public Health Genomics 16, 37–43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Maier AG, Rug M, O'Neill MT, Brown M, Chakravorty S, Szestak T, Chesson J, Wu Y, Hughes K, Coppel RL, Newbold C, Beeson JG, Craig A, Crabb BS and Cowman AF (2008) Exported proteins required for virulence and rigidity of Plasmodium falciparum-infected human erythrocytes. Cell 134, 48–61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Mathema VB and Na-Bangchang K (2015) A brief review on biomarkers and proteomic approach for malaria research. Asian Pacific Journal of Tropical Medicine 8, 253–262. [DOI] [PubMed] [Google Scholar]
  56. Menard D and Dondorp A (2017) Antimalarial drug resistance: a threat to malaria elimination. Cold Spring Harbor Perspectives in Medicine 7, a025619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Miao J, Chen Z, Wang Z, Shrestha S, Li X, Li R and Cui L (2017) Sex-specific biology of the human malaria parasite revealed from the proteomes of mature male and female gametocytes. Molecular & Cellular Proteomics 16, 537–551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Moreno A and Joyner C (2015) Malaria vaccine clinical trials: what's on the horizon. Current Opinion in Immunology 35, 98–106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Mu AK-W, Bee PC, Lau YL and Chen Y (2014) Identification of protein markers in patients infected with Plasmodium knowlesi, Plasmodium falciparum and Plasmodium vivax. International Journal of Molecular Sciences 15, 19952–19961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Obiero JM, Campo JJ, Scholzen A, Randall A, Bijker EM, Roestenberg M, Hermsen CC, Teng A, Jain A, Davies DH, Sauerwein RW and Felgner PL (2019) Antibody biomarkers associated with sterile protection induced by controlled human malaria infection under chloroquine prophylaxis. Msphere 4, e00027–e00019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Ohnishi K and Kimura K (2001) Serum levels of vascular cell adhesion molecule 1 in the early post-treatment defervescent phase of falciparum malaria. Parasitology Research 87, 67–69. [DOI] [PubMed] [Google Scholar]
  62. Ortega C, Frando A, Webb-Robertson B-J, Anderson LN, Fleck N, Flannery EL, Fishbaugher M, Murphree TA, Hansen JR, Smith RD, Kappe SHI, Wright AT and Grundner C (2018) A global survey of ATPase activity in Plasmodium falciparum asexual blood stages and gametocytes. Molecular & Cellular Proteomics 17, 111–120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Ouédraogo A, Tiono AB, Kargougou D, Yaro JB, Ouédraogo E, Kaboré Y, Kangoye D, Bougouma EC, Gansane A, Henri N, Diarra A, Sanon S, Soulama I, Konate AT, Watson NL, Brown V, Hendriks J, Pau MG, Versteege I, Wiesken E, Sadoff J, Nebie I and Sirima SB (2013) A phase 1b randomized, controlled, double-blinded dosage-escalation trial to evaluate the safety, reactogenicity and immunogenicity of an adenovirus type 35 based circumsporozoite malaria vaccine in Burkinabe healthy adults 18 to 45 years of age. PLoS ONE 8, e78679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Pal-Bhowmick I, Mehta M, Coppens I, Sharma S and Jarori GK (2007) Protective properties and surface localization of Plasmodium falciparum enolase. Infection and Immunity 75, 5500–5508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Perera MK, Herath NP, Pathirana SL, Phone-Kyaw M, Alles HK, Mendis KN, Premawansa S and Handunnetti SM (2013) Association of high plasma TNF-alpha levels and TNF-alpha/IL-10 ratios with TNF2 allele in severe P. falciparum malaria patients in Sri Lanka. Pathogens and Global Health 107, 21–29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Ponsford MJ, Medana IM, Prapansilp P, Hien TT, Lee SJ, Dondorp AM, Esiri MM, Day NPJ, White NJ and Turner GDH (2012) Sequestration and microvascular congestion are associated with coma in human cerebral malaria. Journal of Infectious Diseases 205, 663–671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. Qazi KR, Wikman M, Vasconcelos N-M, Berzins K, Ståhl S and Fernández C (2005) Enhancement of DNA vaccine potency by linkage of Plasmodium falciparum malarial antigen gene fused with a fragment of HSP70 gene. Vaccine 23, 1114–1125. [DOI] [PubMed] [Google Scholar]
  68. Ray S, Renu D, Srivastava R, Gollapalli K, Taur S, Jhaveri T, Dhali S, Chennareddy S, Potla A, Dikshit JB, Srikanth R, Gogtay N, Thatte U, Patankar S and Srivastava S (2012) Proteomic investigation of falciparum and vivax malaria for identification of surrogate protein markers. PLoS ONE 7, e41751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  69. Ray S, Kumar V, Bhave A, Singh V, Gogtay NJ, Thatte UM, Talukdar A, Kochar SK, Patankar S and Srivastava S (2015) Proteomic analysis of Plasmodium falciparum induced alterations in humans from different endemic regions of India to decipher malaria pathogenesis and identify surrogate markers of severity. Journal of Proteomics 127, 103–113. [DOI] [PubMed] [Google Scholar]
  70. Reuterswärd P, Bergström S, Orikiiriza J, Lindquist E, Bergström S, Svahn HA, Ayoglu B, Uhlén M, Wahlgren M, Normark J, Ribacke U and Nilsson P (2018) Levels of human proteins in plasma associated with acute paediatric malaria. Malaria Journal 17, 426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  71. Richie TL and Saul A (2002) Progress and challenges for malaria vaccines. Nature 415, 694–701. [DOI] [PubMed] [Google Scholar]
  72. Roestenberg M, McCall M, Hopman J, Wiersma J, Luty AJ, van Gemert GJ, van de Vegte-Bolmer M, van Schaijk B, Teelen K, Arens T, Spaarman L, de Mast Q, Roeffen W, Snounou G, Rénia L, van der Ven A, Hermsen CC and Sauerwein R (2009) Protection against a malaria challenge by sporozoite inoculation. New England Journal of Medicine 361, 468–477. [DOI] [PubMed] [Google Scholar]
  73. Sam-Yellowe TY (2015) Immune complex proteomes: tools for vaccine discovery. Journal of Proteomics & Bioinformatics 8, 188. [Google Scholar]
  74. Sampaio NG, Cheng L and Eriksson EM (2017) The role of extracellular vesicles in malaria biology and pathogenesis. Malaria Journal 16, 245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  75. Sanchez GI, Sedegah M, Rogers WO, Jones TR, Sacci J, Witney A, Carucci DJ, Kumar N and Hoffman SL (2001) Immunogenicity and protective efficacy of a Plasmodium yoelii Hsp60 DNA vaccine in BALB/c mice. Infection and Immunity 69, 3897–3905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  76. Sarfo BO, Hahn A, Schwarz NG, Jaeger A, Sarpong N, Marks F, Adu-Sarkodie Y, Tamminga T and May J (2018) The usefulness of C-reactive protein in predicting malaria parasitemia in a sub-Saharan African region. PLoS ONE 13, e0201693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  77. Seydel KB, Milner DA Jr, Kamiza SB, Molyneux ME and Taylor TE (2006) The distribution and intensity of parasite sequestration in comatose Malawian children. The Journal of Infectious Diseases 194, 208–215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  78. Simon N, Kuehn A, Williamson KC and Pradel G (2016) Adhesion protein complexes of malaria gametocytes assemble following parasite transmission to the mosquito. Parasitology International 65, 27–30. [DOI] [PMC free article] [PubMed] [Google Scholar]
  79. Singh SK, Roeffen W, Andersen G, Bousema T, Christiansen M, Sauerwein R and Theisen M (2015) A Plasmodium falciparum 48/45 single epitope R0. 6C subunit protein elicits high levels of transmission blocking antibodies. Vaccine 33, 1981–1986. [DOI] [PubMed] [Google Scholar]
  80. Siqueira-Batista R, Gomes AP, de Mendonça EG, Vitorino RR, de Azevedo SFM, de Barros Freitas R, Santana LA and Goreti de Almeida Oliveira M (2012) Plasmodium falciparum malaria: proteomic studies. Revista Brasileira de terapia intensiva 24, 394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  81. Souza WD (2006) Secretory organelles of pathogenic protozoa. Anais da Academia Brasileira de Ciências 78, 271–292. [DOI] [PubMed] [Google Scholar]
  82. Suárez-Cortés P, Sharma V, Bertuccini L, Costa G, Bannerman N-L, Sannella AR, Williamson K, Klemba M, Levashina EA, Lasonder E and Alano P (2016) Comparative proteomics and functional analysis reveal a role of Plasmodium falciparum osmiophilic bodies in malaria parasite transmission. Molecular & Cellular Proteomics 15, 3243–3255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  83. Swearingen KE and Lindner SE (2018) Plasmodium parasites viewed through proteomics. Trends in Parasitology 34, 945–960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  84. Swearingen KE, Lindner SE, Shi L, Shears MJ, Harupa A, Hopp CS, Vaughan AM, Springer TA, Moritz RL, Kappe SHI and Sinnis P (2016) Interrogating the Plasmodium sporozoite surface: identification of surface-exposed proteins and demonstration of glycosylation on CSP and TRAP by mass spectrometry-based proteomics. PLoS Pathogens 12, e1005606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  85. Tao D, Ubaida-Mohien C, Mathias DK, King JG, Pastrana-Mena R, Tripathi A, Goldowitz I, Graham DR, Moss E, Marti M and Dinglasan RR (2014) Sex-partitioning of the Plasmodium falciparum stage V gametocyte proteome provides insight into falciparum-specific cell biology. Molecular & Cellular Proteomics 13, 2705–2724. [DOI] [PMC free article] [PubMed] [Google Scholar]
  86. Thera MA, Doumbo OK, Coulibaly D, Laurens MB, Kone AK, Guindo AB, Traore K, Sissoko M, Diallo DA, Diarra I, Kouriba B, Daou M, Dolo A, Baby M, Sissoko MS, Sagara I, Niangaly A, Traore I, Olotu A, Godeaux O, Leach A, Dubois MC, Ballou WR, Cohen J, Thompson D, Dube T, Soisson L, Diggs CL, Takala SL, Lyke KE, House B, Lanar DE, Dutta S, Heppner DG and Plowe CV (2010) Safety and immunogenicity of an AMA1 malaria vaccine in Malian children: results of a phase 1 randomized controlled trial. PLoS ONE 5, e9041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  87. Thima K, Reamtong O, Moonsom S and Chavalitshewinkoon-Petmitr P (2017) Proteomic analysis of asexual stages, young and mature gametocytes of Plasmodium falciparum strain NF54 by mass spectrometry. Southeast Asian Journal of Tropical Medicine and Public Health 48, 711–721. [Google Scholar]
  88. Topolska AE, Richie TL, Nhan DH and Coppel RL (2004) Associations between responses to the rhoptry-associated membrane antigen of Plasmodium falciparum and immunity to malaria infection. Infection and Immunity 72, 3325–3330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  89. Zhang Y, Huang C, Kim S, Golkaram M, Dixon MW, Tilley L, Li J, Zhang S and Suresh S (2015) Multiple stiffening effects of nanoscale knobs on human red blood cells infected with Plasmodium falciparum malaria parasite. Proceedings of the National Academy of Sciences 112, 6068–6073. [DOI] [PMC free article] [PubMed] [Google Scholar]

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