Therapeutic DNA vaccine induces broad T cell responses in the gut and sustained protection from viral rebound and AIDS in SIV-infected rhesus macaques.

Immunotherapies that induce durable immune control of chronic HIV infection may eliminate the need for life-long dependence on drugs. We investigated a DNA vaccine formulated with a novel genetic adjuvant that stimulates immune responses in the blood and gut for the ability to improve therapy in rhesus macaques chronically infected with SIV. Using the SIV-macaque model for AIDS, we show that epidermal co-delivery of plasmids expressing SIV Gag, RT, Nef and Env, and the mucosal adjuvant, heat-labile E. coli enterotoxin (LT), during antiretroviral therapy (ART) induced a substantial 2-4-log fold reduction in mean virus burden in both the gut and blood when compared to unvaccinated controls and provided durable protection from viral rebound and disease progression after the drug was discontinued. This effect was associated with significant increases in IFN-γ T cell responses in both the blood and gut and SIV-specific CD8+ T cells with dual TNF-α and cytolytic effector functions in the blood. Importantly, a broader specificity in the T cell response seen in the gut, but not the blood, significantly correlated with a reduction in virus production in mucosal tissues and a lower virus burden in plasma. We conclude that immunizing with vaccines that induce immune responses in mucosal gut tissue could reduce residual viral reservoirs during drug therapy and improve long-term treatment of HIV infection in humans

Full-length human placental sFlt-1-e15a isoform induces distinct maternal phenotypes of preeclampsia in mice

<div><p>Objective</p><p>Most anti-angiogenic preeclampsia models in rodents utilized the overexpression of a truncated soluble fms-like tyrosine kinase-1 (sFlt-1) not expressed in any species. Other limitations of mouse preeclampsia models included stressful blood pressure measurements and the lack of postpartum monitoring. We aimed to 1) develop a mouse model of preeclampsia by administering the most abundant human placental sFlt-1 isoform (hsFlt-1-e15a) in preeclampsia; 2) determine blood pressures in non-stressed conditions; and 3) develop a survival surgery that enables the collection of fetuses and placentas and postpartum (PP) monitoring.</p><p>Methods</p><p>Pregnancy status of CD-1 mice was evaluated with high-frequency ultrasound on gestational days (GD) 6 and 7. Telemetry catheters were implanted in the carotid artery on GD7, and their positions were verified by ultrasound on GD13. Mice were injected through tail-vein with adenoviruses expressing hsFlt-1-e15a (n = 11) or green fluorescent protein (GFP; n = 9) on GD8/GD11. Placentas and pups were delivered by cesarean section on GD18 allowing PP monitoring. Urine samples were collected with cystocentesis on GD6/GD7, GD13, GD18, and PPD8, and albumin/creatinine ratios were determined. GFP and hsFlt-1-e15a expression profiles were determined by qRT-PCR. Aortic ring assays were performed to assess the effect of hsFlt-1-e15a on endothelia.</p><p>Results</p><p>Ultrasound predicted pregnancy on GD7 in 97% of cases. Cesarean section survival rate was 100%. Mean arterial blood pressure was higher in hsFlt-1-e15a-treated than in GFP-treated mice (∆MAP = 13.2 mmHg, p = 0.00107; GD18). Focal glomerular changes were found in hsFlt-1-e15a -treated mice, which had higher urine albumin/creatinine ratios than controls (109.3±51.7μg/mg vs. 19.3±5.6μg/mg, p = 4.4x10^-2; GD18). Aortic ring assays showed a 46% lesser microvessel outgrowth in hsFlt-1-e15a-treated than in GFP-treated mice (p = 1.2x10^-2). Placental and fetal weights did not differ between the groups. One mouse with liver disease developed early-onset preeclampsia-like symptoms with intrauterine growth restriction (IUGR).</p><p>Conclusions</p><p>A mouse model of late-onset preeclampsia was developed with the overexpression of hsFlt-1-e15a, verifying the <i>in vivo</i> pathologic effects of this primate-specific, predominant placental sFlt-1 isoform. HsFlt-1-e15a induced early-onset preeclampsia-like symptoms associated with IUGR in a mouse with a liver disease. Our findings support that hsFlt-1-e15a is central to the terminal pathway of preeclampsia, and it can induce the full spectrum of symptoms in this obstetrical syndrome.</p></div

Comparison of maternal and umbilical cord blood HIF-1 α

WOS: 000366349300008Objective: Despite the absence of a complete physiologic-pathologic understanding, common accepted theory for development of preeclampsia is incomplete trophoblastic invasion leading to failed uterine and spiral arteriolar remodeling, causing maternal vascular endothelial dysfunction by secreted molecules in response to decreased placental perfusion, placental hypoxia, and ischemia. Placental angiogenesis is especially ineffective in early onset preeclampsia and fetal morbidity/mortality rates are higher because of further decreased blood flow. In this study, we aim to compare the maternal and umbilical cord blood levels of hypoxia-inducible transcription factor-1 alpha (HIF-1 alpha), which is believed to regulate hypoxia-related transcriptional responses, to play role in activating genes for initial phases of placental development and angiogenesis and a physiologic vasodilator molecule nitric oxide (NO) in normal, early and late onset preeclamptic pregnant women. Methods: Pregnant women who were diagnosed with preeclampsia (early onset 34 weeks) and delivered in our clinic were enrolled for this prospective case-controlled study. Pregnant women without preeclampsia were recruited as control group. HIF-1 alpha and NO levels in maternal and umbilical cord blood measured and compared among groups. Findings: A total of 46 cases were enrolled for this study, including 25 preeclamptic (13 in the early onset group and 12 in the late onset group) and 21 normal pregnant women in the control group. Comparison of preeclampsia group to controls revealed higher maternal blood HIF-1 alpha levels in the control group, however higher umbilical cord NO levels in the preeclampsia group (p<0.05 and p<0.001, respectively). In a second analysis, when compared to control group, both early and late onset preeclampsia subgroups were found to have higher umbilical cord blood NO levels (p<0.001). Results: In this study, we observed lower maternal blood HIF-1a levels and higher umbilical cord NO levels in preeclampsia group than controls. These findings suggest that umbilical cord blood NO levels in pregnant women with preeclampsia increase in response to hypoxia. However, lower HIF-1 alpha levels in preeclampsia group can be due to our limited number of cases and we think that there is a need for further studies with larger sample size.Adnan Menderes UniversityAdnan Menderes UniversityThe authors report no conflicts of interest. This research was supported by a research fund from Adnan Menderes University