Genetic manipulation of Anopheles stephensi immunity to increase plasmodium falciparum salivary gland sporozoite infection levels

Human malaria is responsible for over 700,000 deaths a year. To stay abreast of the threat posed by the parasite, a constant stream of new drugs and vector control methods are required. This study focuses on a vaccine that has the potential to protect against parasite infection, but has been hindered by developmental challenges. In malaria prevention, live, attenuated, aseptic, Plasmodium falciparum sporozoites (PfSPZ) can be administered as a highly protective vaccine. PfSPZ are produced using adult female Anopheles stephensi mosquitoes as bioreactors. Production volume and cost of a PfSPZ vaccine for malaria are expected to be directly correlated with Plasmodium falciparum infection intensity in the salivary glands. The sporogonic development of Plasmodium falciparum in A. stephensi to fully infected salivary gland stage sporozoites is dictated by the activities of several known components of the mosquito’s innate immune system. Here I report on the use of genetic technologies that have been rarely, if ever, used in Anopheles stephensi Sda500 to increase the yield of sporozoites per mosquito and enhance vaccine production. By combining the Gal4/UAS bipartite system with in vivo expression of shRNA gene silencing, activity of the IMD signaling pathway downstream effector LRIM1, an antagonist to Plasmodium development, was reduced in the midgut, fat body, and salivary glands of A. stephensi. In infection studies using P. berghei and P. falciparum these transgenic mosquitoes consistently produced significantly more salivary gland stage sporozoites than wildtype controls, with increases in P. falciparum ranging from 2.5 to 10 fold. Using Plasmodium infection assays and qRT-PCR, two novel findings were identified. First, it was shown that 14 days post Plasmodium infection, transcript abundance of the IMD immune effector genes LRIM1, TEP1 and APL1c are elevated, in the salivary glands of A. stephensi, suggesting the salivary glands may play a role in post midgut defense against the parasite. Second, a non-pathogenic IMD signaling pathway response was observed which could suggest an alternative pathway for IMD activation. The information gained from these studies has significantly increased our knowledge of Plasmodium defense in A. stephensi and moreover could significantly improve vaccine production.

RACIAL/ETHNIC VARIABILITY IN THE EFFECT OF RADIATION ON SECOND PRIMARY THYROID CANCER IN CHILDREN: ANALYSIS USING SURVEILLANCE EPIDEMIOLOGY AND END RESULTS DATA

Increases in pediatric thyroid cancer incidence could be partly due to previous clinical intervention. This retrospective cohort study used 1973-2012 data from the Surveillance Epidemiology and End Results program to assess the association between previous radiation therapy exposure in development of second primary thyroid cancer (SPTC) among 0-19-year-old children. Statistical analysis included the calculation of summary statistics and univariable and multivariable logistic regression analysis. Relative to no previous radiation therapy exposure, cases exposed to radiation had 2.46 times the odds of developing SPTC (95% CI: 1.39-4.34). After adjustment for sex and age at diagnosis, Hispanic children who received radiation therapy for a first primary malignancy had 3.51 times the odds of developing SPTC compared to Hispanic children who had not received radiation therapy, [AOR=3.51, 99% CI: 0.69-17.70, p=0.04]. These findings support the development of age-specific guidelines for the use of radiation based interventions among children with and without cancer.