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Orr-Gonzalez, S.

Publications and source records attributed to Orr-Gonzalez, S..

3 recordsLinked to original sources

Aotus nancymaae model predicts human immune response to the placental malaria vaccine candidate VAR2CSA

Placental malaria vaccines (PMV) are being developed to prevent severe sequelae of placental malaria (PM) in pregnant women and their offspring. The leading candidate vaccine antigen VAR2CSA mediates parasite binding to placental receptor chondroitin sulfate A (CSA). Despite promising results in small animal studies, recent human trials of the first two PMV candidates (PAMVAC and PRIMVAC) generated limited cross-reactivity and cross-inhibitory activity to heterologous parasites. Here, we immunized Aotus nancymaae monkeys with three PMV candidates (PAMVAC, PRIMVAC and ID1-ID2a_M1010) adjuvanted with Alhydrogel(R), and exploited the model to investigate boosting of functional vaccine responses during PM episodes as well as with nanoparticle antigens. PMV candidates induced high levels of antigen-specific IgG with significant cross-reactivity across PMV antigens by ELISA. Conversely, PMV antibodies recognized native VAR2CSA and blocked CSA-adhesion of only homologous parasites and not heterologous parasites. PM episodes did not significantly boost VAR2CSA antibody levels or serum functional activity; nanoparticle and monomer antigens alike boosted serum reactivity but not functional activities. Overall, PMV candidates induced functional antibodies with limited heterologous activity in Aotus monkeys, similar to responses reported in humans. The Aotus model appears suitable for preclinical down-selection of PMV candidates and assessment of antibody boosting by PM episodes. Research in Context Evidence before this studyThe Plasmodium falciparum erythrocyte membrane protein VAR2CSA is the leading vaccine candidate antigen to protect pregnant women against placental malaria (PM), which causes serious adverse pregnancy outcomes particularly in first-time mothers living in malaria-endemic areas. Two VAR2CSA-based vaccines (PAMVAC and PRIMVAC) induced strong heterologous functional antibodies in small animals, but induced antibodies with limited cross-inhibitory functional activity in human clinical trials. These observations highlighted the need to establish new animal models that could better recapitulate human pathogenesis and immunity. In ongoing development of a nonhuman primate model for PM, we established an Aotus nancymaae model susceptible to P. falciparum infection during pregnancy that reproduces all the immunoparasitological and histological features of human PM. In this study, we explore the new Aotus model as a platform for evaluating PM vaccine (PMV) immunogenicity and for boosting of vaccine responses during PM episodes. Added value of this studyIn this manuscript, we demonstrate that PMV (including PAMVAC and PRIMVAC) are immunogenic in Aotus monkeys, inducing antibodies with mainly homologous and little heterologous functional activity, as seen in humans but contrary to preclinical reports on these vaccines in small animals. Implications of all the available evidenceOur findings suggest Aotus is a suitable model to assess immunogenicity of VAR2CSA-derived vaccines, in contrast to small animal models. PMV data from human trials and Aotus monkeys suggest that improvements to current VAR2CSA immunogens and/or adjuvants are needed to enhance protective antibody responses, as are studies that evaluate the potential for natural infection to boost vaccine antibody in pregnancy. Therefore, the Aotus PM model may be useful to assess second-generation PMVs seeking to increase strain-transcending activity and to prioritize these for further clinical development.

immunology↗

Design of the SARS-CoV-2 RBD vaccine antigen improves neutralizing antibody response

The receptor binding domain (RBD) of the SARS-CoV-2 spike protein is the primary target of neutralizing antibodies and is a component of almost all vaccine candidates. Here, RBD immunogens were created with stabilizing amino acid changes that improve the neutralizing antibody response, as well as characteristics for production, storage, and distribution. A computational design and in vitro screening platform identified three improved immunogens, each with approximately nine amino acid changes relative to the native RBD sequence and four key changes conserved between immunogens. The changes are adaptable to all vaccine platforms, are compatible with established changes in SARS-CoV-2 vaccines, and are compatible with mutations in emerging variants of concern. The immunogens elicit higher levels of neutralizing antibodies than native RBD, focus the immune response to structured neutralizing epitopes, and have increased production yields and thermostability. Incorporating these variant-independent amino acid changes in next-generation vaccines may enhance the neutralizing antibody response and lead to pan-SARS-CoV-2 protection.

molecular biology↗

Activity of Plasmodium vivax promoter elements in Plasmodium knowlesi, and a centromere-containing plasmid that expresses NanoLuc throughout the parasite life cycle

BackgroundPlasmodium knowlesi is now the major cause of human malaria in Malaysia, complicating malaria control efforts that must attend to the elimination of multiple Plasmodium species. Recent advances in the cultivation of P. knowlesi erythrocytic-stage parasites in vitro, transformation with exogenous DNA, and infection of mosquitoes with gametocytes from culture have opened up studies of this pathogen without the need for resource-intensive and costly non-human primate (NHP) models. For further understanding and development of methods for parasite transformation in malaria research, this study examined the activity of various trans-species transcriptional control sequences and the influence of Plasmodium vivax centromeric (pvcen) repeats in plasmid-transfected P. knowlesi parasites. MethodsIn vitro cultivated P. knowlesi parasites were transfected with plasmid constructs that incorporated P. vivax or Plasmodium falciparum 5 UTRs driving the expression of bioluminescence markers (firefly luciferase or Nanoluc). Promoter activities were assessed by bioluminescence, and parasites transformed with human resistant allele dihydrofolate reductase-expressing plasmids were selected using antifolates. The stability of transformants carrying pvcen-stabilized episomes was assessed by bioluminescence over a complete parasite life cycle through a rhesus macaque monkey, mosquitoes, and a second rhesus monkey. ResultsLuciferase expression assessments show that certain P. vivax promoter regions, not functional in the more evolutionarily-distant P. falciparum, can drive transgene expression in P. knowlesi. Further, pvcen repeats may improve the stability of episomal plasmids in P. knowlesi and support detection of NanoLuc-expressing elements over the full parasite life cycle from rhesus macaque monkeys to Anopheles dirus mosquitoes and back again to monkeys. In assays of drug responses to chloroquine, G418 and WR9910, antimalarial half-inhibitory concentration (IC50) values of blood stages measured by NanoLuc activity proved comparable to IC50 values measured by the standard SYBR Green method. ConclusionAll three P. vivax promoters tested in this study functioned in P. knowlesi whereas two of the three were inactive in P. falciparum. NanoLuc-expressing, centromere-stabilized plasmids may support high-throughput screenings of P. knowlesi for new antimalarial agents, including compounds that can block the development of mosquito- and/or liver-stage parasites.

microbiology↗