scholarly journals Orientation of Antigen Display on Self-Assembling Protein Nanoparticles Influences Immunogenicity

Vaccines ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 103
Author(s):  
Cosette G. Schneider ◽  
Justin A. Taylor ◽  
Michael Q. Sibilo ◽  
Kazutoyo Miura ◽  
Katherine L. Mallory ◽  
...  
Keyword(s):  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Effective Vaccine ◽  
Antigen Delivery ◽  
Self Assembling ◽  
Protein Nanoparticles ◽  
Delivery Platform ◽  
Dual Stage ◽  
Functional Antibodies ◽  
Stage Parasite

Self-assembling protein nanoparticles (SAPN) serve as a repetitive antigen delivery platform with high-density epitope display; however, antigen characteristics such as size and epitope presentation can influence the immunogenicity of the assembled particle and are aspects to consider for a rationally designed effective vaccine. Here, we characterize the folding and immunogenicity of heterogeneous antigen display by integrating (a) dual-stage antigen SAPN presenting the P. falciparum (Pf) merozoite surface protein 1 subunit, PfMSP119, and Pf cell-traversal protein for ookinetes and sporozoites, PfCelTOS, in addition to (b) a homogenous antigen SAPN displaying two copies of PfCelTOS. Mice and rabbits were utilized to evaluate antigen-specific humoral and cellular induction as well as functional antibodies via growth inhibition of the blood-stage parasite. We demonstrate that antigen orientation and folding influence the elicited immune response, and when appropriately designed, SAPN can serve as an adaptable platform for an effective multi-antigen display.

scholarly journals Genetic Characterization of Plasmodium vivax Isolates from Pakistan Using Circumsporozoite Protein (PvCSP) and Merozoite Surface Protein-1 (PvMSP-1) genes as genetic markers 

2021 ◽  
Author(s):  
Zainab Bibi ◽  
Anam Fatima ◽  
Rehana Rani ◽  
Ayesha Maqbool ◽  
Samea Khan ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Plasmodium Vivax ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Circumsporozoite Protein ◽  
Effective Vaccine ◽  
Population Divergence ◽  
Large Sample Size ◽  
Base Line ◽  
Merozoite Surface Protein 1

Abstract Background: Plasmodium vivax contributes to over 70% malaria burden in Pakistan, but limited data exists on various aspects including genetic diversity of the parasite as compared to other parts of the world. Since the information about the genetic diversity of P. vivax assists to understand the population dynamics of the parasite, the current study was designed to understand population divergence of Plasmodium vivax in Pakistan using circumsporozoite protein (PvCSP) and merozoite surface protein-1 (PvMSP-1) genes as molecular markers. Methods: PvCSP and PvMSP-1 specific PCR and DNA sequencing were carried out for 150 blood samples collected from Islamabad and Rawalpindi, Pakistan. Genetic diversity and polymorphism was analyzed using ChromasPro, ClustalW, MEGA7, DnaSP v.5 and WebLogo programs. Results: The PCR for PvCSP and PvMSP-1 genes was carried out for 150 P. vivax isolates and resulting the PCR products ranging from 900 to 1100 bp for PvCSP and ~400bp for PvMSP-1 genes respectively. Majority (93%; 141/150) of the P. vivax isolates were of VK210 variant and only 9 isolates were found to be of VK247 variant based on PvCSP gene. Out of the numerous peptide repeat motifs (PRMs) detected, GDRADGQPA (PRM1) and GDRAAGQPA (PRM2) were more extensively dispersed among the P. vivax isolates. Partial sequences (~400bp) at the N-terminal of PvMSP-1 gene depicted high level of diversity.Conclusion: High levels of genetic diversity based on PvCSP and PvMSP-1 genes was observed in the isolated samples from the study area. Parasite typing is essential in predicting pattern of antigenic variations and drug resistance and for effective vaccine designing and development which can further assist in evaluating measures for malaria control at individual and community level. The base-line data presented here warrants future studies to investigate more into the genetic diversity of P. vivax with large sample size from across the country for better understanding of the transmission patterns of vivax malaria.

scholarly journals Immunogenicity analysis of conserved fragments in Plasmodium ovale species merozoite surface protein 4

2020 ◽  
Author(s):  
Uwase Juliette ◽  
Ruilin Chu ◽  
Kokouvi Kassegne ◽  
Yao Lei ◽  
Feihu Shen ◽  
...  
Keyword(s):  
Immune Responses ◽  
Plasmodium Species ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Effective Vaccine ◽  
Plasmodium Ovale ◽  
Tropical Regions ◽  
Cellular Immune Responses ◽  
Cellular Immune ◽  
Proliferation Assays

Abstract Background: There is an urgent need for an effective vaccine to control and eradicate malaria, one of the most serious global infectious diseases. Plasmodium merozoite surface protein 4 (MSP4) has been listed as a blood-stage subunit vaccine candidate for malaria. Plasmodium ovale species (spp.) infection is also a source of malaria burden in tropical regions where it is sometimes mixed with other Plasmodium species. However, little is known about P. ovale spp. MSP4. Methods: The msp4 gene was amplified through polymerase chain reaction using genomic DNA extracted from blood samples of 46 patients infected with P. ovale spp. and amplified products were sequenced. Open reading frames predicted as immunogenic peptides consisting of 119 and 97 amino acids of P. ovale curtisi MSP4 (PocMSP4) and P. ovale wallikeri MSP4 (PowMSP4), respectively, were selected for protein expression. Recombinant proteins (rPoMSP4) were expressed in Escherichia coli, purified, analyzed, and immunized in BALB/c mice. The specificity of anti-MSP4-immunoglubulin (Ig) G antibodies was evaluated by Western blot and enzyme-linked immunosorbent assays, and cellular immune responses were analyzed via lymphocyte proliferation assays. Results: Full peptide sequences of PocMSP4 and PowMSP4 were completely conserved in all clinical isolates, except in the epidermal growth factor-like domain at the carboxyl terminus where only one mutation was observed in one P. ovale wallikeri isolate. Furthermore, we successfully expressed the truncated pocmsp4 and powmsp4 and purified as ~32 kDa proteins. Our results showed that PocMSP4 and PowMSP4 induced high antibody responses with end-point titers ranging from 1:10,000 to 1:2,560,000 in all immunized mouse groups and with high IgG avidity to PocMSP4 (80.5%) and PowMSP4 (92.3%). Furthermore, rPocMSP4 and rPowMSP4 cross-reacted with anti-PowMSP4-specific or anti-PocMSP4-specific antibodies. Additionally, anti-PoMSP4 IgG antibodies showed broad immuno-specificity in reacting against rPoMSP1 and rPoAMA1. Lastly, PocMSP4- and PowMSP4-immunized mice induced cellular immune responses with PocMSP4 (36%) and PowMSP4 cells (15.8%) as observed through splenocyte proliferation assays. Conclusion: Our study suggested conservation in PoMSP4 protein sequences and high immunogenicity was observed in rPoMSP4. Furthermore, PocMSP4- and PowMSP4-immunized mice induced cellular immune responses, suggesting that both humoral and cellular immune responses play crucial roles in the protection against any antigen.

scholarly journals Malaria Immunoepidemiology in Low Transmission: Correlation of Infecting Genotype and Immune Response to Domains of Plasmodium falciparum Merozoite Surface Protein 3

2011 ◽  
Vol 79 (5) ◽  
pp. 2070-2078 ◽  
Author(s):  
Stephen J. Jordan ◽  
Ana L. Oliveira ◽  
Jean N. Hernandez ◽  
Robert A. Oster ◽  
Debasish Chattopadhyay ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Effective Vaccine ◽  
Serum Samples ◽  
Content Type ◽  
Low Transmission ◽  
Rational Development ◽  
Time Of Infection ◽  
Potential Vaccine

ABSTRACTMalaria caused byPlasmodium falciparumis a major cause of global infant mortality, and no effective vaccine currently exists. Multiple potential vaccine targets have been identified, and immunoepidemiology studies have played a major part in assessing those candidates. When such studies are carried out in high-transmission settings, individuals are often superinfected with complex mixtures of genetically distinctP. falciparumtypes, making it impossible to directly correlate the genotype of the infecting antigen with the antibody response. In contrast, in regions of low transmissionP. falciparuminfections are often genetically simple, and direct comparison of infecting genotype and antigen-specific immune responses is possible. As a test of the utility of this approach, responses against several domains and allelic variants of the vaccine candidateP. falciparummerozoite surface protein 3 (PfMSP3) were tested in serum samples collected near Iquitos, Peru. Antibodies recognizing both the conserved C-terminal and the more variable N-terminal domain were identified, but anti-N-terminal responses were more prevalent, of higher titers, and primarily of cytophilic subclasses. Comparing antibody responses to different PfMSP3 variants with thePfMSP3genotype present at the time of infection showed that anti-N-terminal responses were largely allele class specific, but there was some evidence for responses that cross-reacted across allele classes. Evidence for cross-reactive responses was much stronger when variants within one allele class were tested, which has implications for the rational development of genotype-transcending PfMSP3-based vaccines.

Immunogenicity analysis of conserved fragments in Plasmodium ovale merozoite surface protein 4

2020 ◽  
Author(s):  
Uwase Juliette ◽  
Ruilin Chu ◽  
Kokouvi Kassegne ◽  
Yao Lei ◽  
Feihu Shen ◽  
...  
Keyword(s):  
Immune Responses ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Effective Vaccine ◽  
Plasmodium Ovale ◽  
Tropical Regions ◽  
Malaria Burden ◽  
Cellular Immune Responses ◽  
Cellular Immune ◽  
Proliferation Assays

Abstract Background: An effective vaccine to control and eradicate malaria, one of the most serious global infectious diseases, is an urgent need. Plasmodium merozoite surface protein 4 (MSP4) has been listed as a blood-stage subunit vaccine candidate for malaria. Plasmodium ovale infection is also a source of malaria burden in tropical regions where it is sometimes mixed with other Plasmodium species. However, little is known about P. ovale MSP4. Methods: The msp4 gene was amplified through polymerase chain reaction using genomic DNA extracted from blood samples of 46 patients infected with P. ovale and then sequenced. Open reading frames predicted as immunogenic peptides consisting of 119 and 97 amino acids of P. ovale curtisi MSP4 (PocMSP4) and P. ovale wallikeri MSP4 (PowMSP4), respectively, were selected for protein expression. Recombinant proteins (rPoMSP4) were expressed in Escherichia coli, purified, analyzed, and immunized in BALB/c mice. The specificity of anti-MSP4-immunoglubulin (Ig) G antibodies was evaluated by Western blot and enzyme-linked immunosorbent assays, and cellular immune responses were analyzed via lymphocyte proliferation assays.Results: Full peptide sequences of PocMSP4 and PowMSP4 were completely conserved in all clinical isolates, except in the epidermal growth factor-like domain at the carboxyl terminus where only one mutation was observed in one P. ovale wallikeri isolate. Furthermore, we successfully expressed the truncated pocmsp4 and powmsp4 and purified as ~32 kDa proteins. Our results showed that PocMSP4 and PowMSP4 induced high antibody responses with end-point titers ranging from 1:2,560,000 to 1:10,000 in all immunized mouse groups and with high IgG avidity to PocMSP4 (80.5%) and PowMSP4 (92.3%). Furthermore, rPocMSP4 and rPowMSP4 cross-reacted with anti-PowMSP4-specific or anti-PocMSP4-specific antibodies. Additionally, anti-PoMSP4 IgG antibodies showed broad immuno-specificity in reacting against rPoMSP1 and rPoAMA1. Lastly, PocMSP4- and PowMSP4-immunized mice induced cellular immune responses with PocMSP4 (36%) and PowMSP4 cells (15.8%) as observed through splenocyte proliferation assays. Conclusion: Our study suggested conservation in PoMSP4 protein sequences with high immunogenicity. Furthermore, PocMSP4- and PowMSP4-immunized mice induced cellular immune responses, suggesting that both humoral and cellular immune responses play crucial roles in the protection against any antigen.

scholarly journals Contrasting Patterns of Serologic and Functional Antibody Dynamics to Plasmodium falciparum Antigens in a Kenyan Birth Cohort

2015 ◽  
Vol 23 (2) ◽  
pp. 104-116 ◽  
Author(s):  
Arlene E. Dent ◽  
Indu Malhotra ◽  
Xuelie Wang ◽  
Denise Babineau ◽  
Kee Thai Yeo ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Cord Blood ◽  
Natural Infection ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
In Utero ◽  
Content Type ◽  
Malaria Exposure ◽  
Increased Risk ◽  
Functional Antibodies

ABSTRACTIgG antibodies toPlasmodium falciparumare transferred from the maternal to fetal circulation during pregnancy, wane after birth, and are subsequently acquired in response to natural infection. We examined the dynamics of malaria antibody responses of 84 Kenyan infants from birth to 36 months of age by (i) serology, (ii) variant surface antigen (VSA) assay, (iii) growth inhibitory activity (GIA), and (iv) invasion inhibition assays (IIA) specific for merozoite surface protein 1 (MSP1) and sialic acid-dependent invasion pathway. Maternal antibodies in each of these four categories were detected in cord blood and decreased to their lowest level by approximately 6 months of age. Serologic antibodies to 3 preerythrocytic and 10 blood-stage antigens subsequently increased, reaching peak prevalence by 36 months. In contrast, antibodies measured by VSA, GIA, and IIA remained low even up to 36 months. Infants sensitized toP. falciparum in utero, defined by cord blood lymphocyte recall responses to malaria antigens, acquired antimalarial antibodies at the same rate as those who were not sensitizedin utero, indicating that fetal exposure to malaria antigens did not affect subsequent infant antimalarial responses. Infants with detectable serologic antibodies at 12 months of age had an increased risk ofP. falciparuminfection during the subsequent 24 months. We conclude that serologic measures of antimalarial antibodies in children 36 months of age or younger represent biomarkers of malaria exposure rather than protection and that functional antibodies develop after 36 months of age in this population.

scholarly journals Detection of High Frequency of MAD20 Allelic Variants of Plasmodium Falciparum Merozoite Surface Protein 1 Gene from Adama and its Surroundings, Oromia, Ethiopia

2021 ◽  
Author(s):  
Temesgen File ◽  
Tsegaye Chekol ◽  
Gezahegn Solomon ◽  
Hunduma Dinka ◽  
Lemu Golassa
Keyword(s):  
Plasmodium Falciparum ◽  
Genetic Polymorphism ◽  
Malaria Control ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Allelic Variant ◽  
Clinical Isolates ◽  
Effective Vaccine ◽  
Multiple Infections ◽  
Size Polymorphism

Abstract Background: One of the major challenges in developing an effective vaccine against asexual stages of Plasmodium falciparum is genetic polymorphism within parasite population. Understanding the genetic polymorphism like block 2 region of merozoite surface protein (msp-1) genes of P. falciparum enlighten mechanisms underlining disease pathology, identification of the parasite clone profile from the isolates, transmission intensity and potential deficiencies of the ongoing malaria control and elimination effort in the locality. Detailed understanding of local genetic polymorphism is an input to pave the way for better management, control and elimination of malaria. The aim of this study was to detect the most frequent allelic variant of the merozoite surface protein (msp-1) gene of P. falciparum clinical isolates from selected health facilities in Adama town and its surroundings, Oromia, Ethiopia.Methods: A total of 139 clinical isolates were successfully amplified for msp-1 gene using specific sets of primer. Nested PCR amplification conducted, using specific primers targeting K1, MAD20, and R033 alleles followed by gel electrophoresis for fragment analysis. Based on the detection of a PCR fragment, infections were classified as monoclonal or multiple infections.Result: 19 different size polymorphism of msp-1 gene were identified in the study, with 67(48 %) MAD20, 18 (13 %) K-1 and 18 (13 %) RO33 allelic family. Whereas, the multiple infections were 21(15 %), 8(5.8 %), 4(2.9 %), 3(2.2 %) for MAD20+K-1, MAD20+RO33, K-1+ RO33, and MAD20+K-1, RO33, respectively. The overall Multiplicity of Infection (MOI) was 1.3 and the expected heterozygosity (He) was 0.58 indicating intermediate falciparum malaria transmission.Conclusion: The status of msp-1 allele size polymorphism, MOI and He observed in the study revealed an intermediate genetic diversity of P. falciparum clinical isolates, indicating that the ongoing malaria control and elimination effort should be intensified to effectively monitor the potential malaria resurgence in the study area. Moreover, deriving force that led to high predominance of MAD20 allelic variant revealed in such malaria declining region demands further research.

scholarly journals Detection of high frequency of MAD20 allelic variants of Plasmodium falciparum merozoite surface protein 1 gene from Adama and its surroundings, Oromia, Ethiopia

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Temesgen File ◽  
Tsegaye Chekol ◽  
Gezahegn Solomon ◽  
Hunduma Dinka ◽  
Lemu Golassa
Keyword(s):  
Plasmodium Falciparum ◽  
Genetic Polymorphism ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Allelic Variant ◽  
Clinical Isolates ◽  
Effective Vaccine ◽  
Multiple Infections ◽  
Size Polymorphism ◽  
Merozoite Surface Protein 1

Abstract Background One of the major challenges in developing an effective vaccine against asexual stages of Plasmodium falciparum is genetic polymorphism within parasite population. Understanding the genetic polymorphism like block 2 region of merozoite surface protein-1 (msp-1) gene of P. falciparum enlighten mechanisms underlining disease pathology, identification of the parasite clone profile from the isolates, transmission intensity and potential deficiencies of the ongoing malaria control and elimination efforts in the locality. Detailed understanding of local genetic polymorphism is an input to pave the way for better management, control and elimination of malaria. The aim of this study was to detect the most frequent allelic variant of the msp-1 gene of P. falciparum clinical isolates from selected health facilities in Adama town and its surroundings, Oromia, Ethiopia. Methods One hundred thirty-nine clinical isolates were successfully amplified for msp-1 gene using specific primers. Nested PCR amplification was conducted targeting K1, MAD20, and R033 alleles followed by gel electrophoresis for fragment analysis. Based on the detection of a PCR fragment, infections were classified as monoclonal or multiple infections. Results 19 different size polymorphism of msp-1 gene were identified in the study, with 67(48%) MAD20, 18 (13%) K-1 and 18 (13%) RO33 allelic family. Whereas, the multiple infections were 21(15%), 8 (5.8%), 4(2.9%), 3(2.2%) for MAD20 + K-1, MAD20 + RO33, K-1 + RO33, and MAD20 + K-1, RO33, respectively. The overall Multiplicity of infection (MOI) was 1.3 and the expected heterozygosity (He) was 0.39 indicating slightly low falciparum malaria transmission. Conclusion The status of msp-1 allele size polymorphism, MOI and He observed in the study revealed the presence of slightly low genetic diversity of P. falciparum clinical isolates. However, highly frequent MAD20 allelic variant was detected from clinical isolates in the study area. Moreover, the driving force that led to high predominance of MAD20 allelic variant revealed in such malaria declining region demands further research.

scholarly journals Immunogenicity analysis of conserved fragments in Plasmodium ovale species merozoite surface protein 4

2020 ◽  
Author(s):  
Juliette Uwase ◽  
Ruilin Chu ◽  
Kokouvi Kassegne ◽  
Yao Lei ◽  
Feihu Shen ◽  
...  
Keyword(s):  
Immune Responses ◽  
Plasmodium Species ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Effective Vaccine ◽  
Plasmodium Ovale ◽  
Tropical Regions ◽  
Cellular Immune Responses ◽  
Cellular Immune ◽  
Proliferation Assays

Abstract Background : There is an urgent need for an effective vaccine to control and eradicate malaria, one of the most serious global infectious diseases. Plasmodium merozoite surface protein 4 (MSP4) has been listed as a blood-stage subunit vaccine candidate for malaria. Plasmodium ovale species (spp.) infection is also a source of malaria burden in tropical regions where it is sometimes mixed with other Plasmodium species. However, little is known about P. ovale spp. MSP4. Methods : The msp4 gene was amplified through polymerase chain reaction using genomic DNA extracted from blood samples of 46 patients infected with P. ovale spp. and amplified products were sequenced. Open reading frames predicted as immunogenic peptides consisting of 119 and 97 amino acids of P. ovale curtisi MSP4 (PocMSP4) and P. ovale wallikeri MSP4 (PowMSP4), respectively, were selected for protein expression. Recombinant proteins (rPoMSP4) were expressed in Escherichia coli , purified, analyzed, and immunized in BALB/c mice. The specificity of anti-MSP4-immunoglubulin (Ig) G antibodies was evaluated by Western blot and enzyme-linked immunosorbent assays, and cellular immune responses were analyzed via lymphocyte proliferation assays. Results : Full peptide sequences of PocMSP4 and PowMSP4 were completely conserved in all clinical isolates, except in the epidermal growth factor-like domain at the carboxyl terminus where only one mutation was observed in one P. ovale wallikeri isolate. Furthermore, we successfully expressed the truncated pocmsp4 and powmsp4 and purified as ~32 kDa proteins. Our results showed that PocMSP4 and PowMSP4 induced high antibody responses with end-point titers ranging from 1:10,000 to 1:2,560,000 in all immunized mouse groups and with high IgG avidity to PocMSP4 (80.5%) and PowMSP4 (92.3%). Furthermore, rPocMSP4 and rPowMSP4 cross-reacted with anti-PowMSP4-specific or anti-PocMSP4-specific antibodies. Additionally, anti-PoMSP4 IgG antibodies showed broad immuno-specificity in reacting against rPoMSP1 and rPoAMA1. Lastly, PocMSP4- and PowMSP4-immunized mice induced cellular immune responses with PocMSP4 (36%) and PowMSP4 cells (15.8%) as observed through splenocyte proliferation assays. Conclusion : Our study suggested conservation in PoMSP4 protein sequences and high immunogenicity was observed in rPoMSP4. Furthermore, PocMSP4- and PowMSP4-immunized mice induced cellular immune responses, suggesting that both humoral and cellular immune responses play crucial roles in the protection against any antigen.

scholarly journals PvMSP8 as a Novel Plasmodium vivax Malaria Sero-Marker for the Peruvian Amazon

Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 282
Author(s):  
Elizabeth Villasis ◽  
Katherine Garro ◽  
Angel Rosas-Aguirre ◽  
Pamela Rodriguez ◽  
Jason Rosado ◽  
...  
Keyword(s):  
Plasmodium Vivax ◽  
Area Under The Curve ◽  
Surface Protein ◽  
Merozoite Surface Protein ◽  
Population Based ◽  
Peruvian Amazon ◽  
Cross Sectional Survey ◽  
Cross Sectional ◽  
Malaria Exposure ◽  
Plasmodium Vivax Malaria

The measurement of recent malaria exposure can support malaria control efforts. This study evaluated serological responses to an in-house Plasmodium vivax Merozoite Surface Protein 8 (PvMSP8) expressed in a Baculovirus system as sero-marker of recent exposure to P. vivax (Pv) in the Peruvian Amazon. In a first evaluation, IgGs against PvMSP8 and PvMSP10 proteins were measured by Luminex in a cohort of 422 Amazonian individuals with known history of Pv exposure (monthly data of infection status by qPCR and/or microscopy over five months). Both serological responses were able to discriminate between exposed and non-exposed individuals in a good manner, with slightly higher performance of anti-PvMSP10 IgGs (area under the curve AUC = 0.78 [95% CI = 0.72–0.83]) than anti-PvMSP8 IgGs (AUC = 0.72 [95% CI = 0.67–0.78]) (p = 0.01). In a second evaluation, the analysis by ELISA of 1251 plasma samples, collected during a population-based cross-sectional survey, confirmed the good performance of anti-PvMSP8 IgGs for discriminating between individuals with Pv infection at the time of survey and/or with antecedent of Pv in the past month (AUC = 0.79 [95% CI = 0.74–0.83]). Anti-PvMSP8 IgG antibodies can be considered as a good biomarker of recent Pv exposure in low-moderate transmission settings of the Peruvian Amazon.

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