Prompt and precise identification of various sources of infection in response to the prevention of malaria re-establishment in China.

Prompt and precise diagnosis of patients is an essential component of malaria control and elimination strategies, it is even more vital for the prevention of malaria re-establishment in the post elimination phase. After eliminating malaria in China, the strategy for prevention of malaria re-establishment was updated in a timely manner from the elimination strategy focusing on each case/focus to the prevention of re-establishment focusing on timely identification of the source of infection. However, there are numerous challenges, such as the persistent large number of imported malaria cases, the long-term threat of border malaria, unknown levels of asymptomatic infections and Plasmodium falciparum HRP2/3 gene deletions, and the continuous spreading of antimalarial drug resistance. Meanwhile, the detection capacity also need to be further improved to meet the timely detection of all sources of infection, otherwise it is bound to occur introduced malaria cases and malaria re-establishment in the presence of malaria vector mosquitoes. Therefore, it is necessary to continuously strengthen the malaria detection competency at all levels, promote the research and development on the malaria parasitological testing technologies, thus improving the timely detection of various sources of infection, and preventing the re-establishment of malaria.

Systems analysis of immune responses to attenuated P. falciparum malaria sporozoite vaccination reveals excessive inflammatory signatures correlating with impaired immunity.

Immunization with radiation-attenuated sporozoites (RAS) can confer sterilizing protection against malaria, although the mechanisms behind this protection are incompletely understood. We performed a systems biology analysis of samples from the Immunization by Mosquito with Radiation Attenuated Sporozoites (IMRAS) trial, which comprised P. falciparum RAS-immunized (PfRAS), malaria-naive participants whose protection from malaria infection was subsequently assessed by controlled human malaria infection (CHMI). Blood samples collected after initial PfRAS immunization were analyzed to compare immune responses between protected and non-protected volunteers leveraging integrative analysis of whole blood RNA-seq, high parameter flow cytometry, and single cell CITEseq of PBMCs. This analysis revealed differences in early innate immune responses indicating divergent paths associated with protection. In particular, elevated levels of inflammatory responses early after the initial immunization were detrimental for the development of protective adaptive immunity. Specifically, non-classical monocytes and early type I interferon responses induced within 1 day of PfRAS vaccination correlated with impaired immunity. Non-protected individuals also showed an increase in Th2 polarized T cell responses whereas we observed a trend towards increased Th1 and T-bet+ CD8 T cell responses in protected individuals. Temporal differences in genes associated with natural killer cells suggest an important role in immune regulation by these cells. These findings give insight into the immune responses that confer protection against malaria and may guide further malaria vaccine development. Trial registration: ClinicalTrials.gov NCT01994525.

Transient knockdown of Anopheles stephensi LRIM1 using RNAi increases Plasmodium falciparum sporozoite salivary gland infections.

BACKGROUND: Plasmodium falciparum (Pf) sporozoites (PfSPZ) can be administered as a highly protective vaccine conferring the highest protection seen to date. Sanaria® PfSPZ vaccines are produced using aseptically reared Anopheles stephensi mosquitoes. The bionomics of sporogonic development of P. falciparum in A. stephensi to fully mature salivary gland PfSPZ is thought to be modulated by several components of the mosquito innate immune system. In order to increase salivary gland PfSPZ infections in A. stephensi and thereby increase vaccine production efficiency, a gene knock down approach was used to investigate the activity of the immune deficiency (IMD) signaling pathway downstream effector leucine-rich repeat immune molecule 1 (LRIM1), an antagonist to Plasmodium development. METHODS: Expression of LRIM1 in A. stephensi was reduced following injection of double stranded (ds) RNA into mosquitoes. By combining the Gal4/UAS bipartite system with in vivo expression of short hairpin (sh) RNA coding for LRIM1 reduced expression of LRIM1 was targeted in the midgut, fat body, and salivary glands. RT-qPCR was used to demonstrate fold-changes in gene expression in three transgenic crosses and the effects on P. falciparum infections determined in mosquitoes showing the greatest reduction in LRIM1 expression. RESULTS: LRIM1 expression could be reduced, but not completely silenced, by expression of LRIM1 dsRNA. Infections of P. falciparum oocysts and PfSPZ were consistently and significantly higher in transgenic mosquitoes than wild type controls, with increases in PfSPZ ranging from 2.5- to tenfold. CONCLUSIONS: Plasmodium falciparum infections in A. stephensi can be increased following reduced expression of LRIM1. These data provide the springboard for more precise knockout of LRIM1 for the eventual incorporation of immune-compromised A. stephensi into manufacturing of Sanaria's PfSPZ products.

Mathematical Analysis of the Ross-Macdonald Model with Quarantine.

People infected with malaria may receive less mosquito bites when they are treated in well-equipped hospitals or follow doctors' advice for reducing exposure to mosquitoes at home. This quarantine-like intervention measure is especially feasible in countries and areas approaching malaria elimination. Motivated by mathematical models with quarantine for directly transmitted diseases, we develop a mosquito-borne disease model where imperfect quarantine is considered to mitigate the disease transmission from infected humans to susceptible mosquitoes. The basic reproduction number [Formula: see text] is computed and the model equilibria and their stabilities are analyzed when the incidence rate is standard or bilinear. In particular, the model system may undergo a subcritical (backward) bifurcation at [Formula: see text] when standard incidence is adopted, whereas the disease-free equilibrium is globally asymptotically stable as [Formula: see text] and the unique endemic equilibrium is locally asymptotically stable as [Formula: see text] when the infection incidence is bilinear. Numerical simulations suggest that the quarantine strategy can play an important role in decreasing malaria transmission. The success of quarantine mainly relies on the reduction of bites on quarantined individuals.