Preferential Targeting of Conserved Gag Regions after Vaccination with a Heterologous DNA prime - Modified Vaccinia Ankara (MVA) boost HIV-1 vaccine regimen

Prime-boost vaccination strategies against HIV-1 often include multiple variants for a given immunogen for better coverage of the extensive viral diversity. To study the immunologic effects of this approach, we characterized breadth, phenotype, function and specificity of Gag-specific T cells induced by a DNA-prime Modified Vaccinia Ankara (MVA)-boost vaccination strategy, which uses mismatched Gag immunogens in the TamoVac 01 phase IIa trial. Healthy Tanzanian volunteers received three injections of the DNA-SMI vaccine encoding for a subtype B and AB-recombinant Gagp37 and two vaccinations with MVA-CMDR encoding subtype A Gagp55 Gag-specific T-cell responses were studied in 42 vaccinees using fresh peripheral blood mononuclear cells. After the first MVA-CMDR boost, vaccine-induced IFN-γ(+) Gag-specific T cell responses were dominated by CD4(+) T cells (compared to CD8(+) T cells, p<0.001) that co-expressed IL-2 (66.4%) and/or TNFα (63.7%). A median of 3 antigenic regions were targeted with a higher median response magnitude to Gagp24 regions - more conserved between prime and boost - as compared to regions within Gagp15 (not primed) and Gagp17 (less conserved, both p<0.0001). Four regions within Gagp24 were each targeted by 45% to 74% of vaccinees upon restimulation with DNA-SMI-Gag matched peptides. The response rate to individual antigenic regions correlated with the sequence homology between the MVA and DNA Gag encoded immunogens (p=0.04, r(2)=0.47). In summary, after the first MVA-CMDR boost, the sequence-mismatched DNA-prime MVA-boost vaccine strategy induced a Gag-specific T cell response that was dominated by polyfunctional CD4(+) T cells and that targeted multiple antigenic regions within the conserved Gagp24 Protein.IMPORTANCE Genetic diversity is a major challenge for the design of vaccines against variable viruses. While including multiple variants for a given immunogen in prime-boost vaccination strategies is one approach that aims to improve coverage for global virus variants, the immunologic consequences of this strategy have been poorly defined so far. It is unclear whether inclusion of multiple variants in prime-boost vaccination strategies improves recognition of variant viruses by T cells and by which mechanisms this would be achieved; either by improved cross-recogniton of multiple variants for a given antigenic region or rather through preferential targeting of antigenic regions more conserved between prime and boost. Engineering vaccines to induce adaptive immune responses that preferentially target conserved antigenic regions of viral vulnerability might facilitate better immune control after preventive and therapeutic vaccination for HIV and for other variable viruses

Appraisal of health care: from patient value to societal benefit

Aim: This paper summarizes the deficiencies and weaknesses of the most frequently used methods for the allocation of health-care resources. New, more transparent and practical methods for optimizing the allocation of these resources are proposed. Method: The examples of quality-adjusted life years (QALYs) and efficiency frontier (EF) are analyzed to describe weaknesses and problems in decisions regulating health-care provision. After conducting a literature search and discussions with an international group of professionals, three groups of professionals were formed to discuss the assessment and appraisal of health-care services and allocation of available resources. Results: At least seven essential variables were identified that should be heeded when applying the concept of QALYs for decisions concerning health-care provision. The efficiency frontier (EF) concept can be used to set a ceiling price and perform a cost-benefit analysis of provision, but different stakeholders—a biostatistician (efficacy), an economist (costs), a clinician (effectiveness), and the patient (value)—could provide a fairer appraisal of health-care services. Efficacy and costs are often based on falsifiable data. Effectiveness and value depend on the success with which a particular clinical problem has been solved. These data cannot be falsified. The societal perspective is generated by an informal cost-benefit analysis including appraisals by the above-mentioned stakeholders and carried out by an authorized institution. Conclusion: Our analysis suggests that study results expressed in QALYs or as EF cannot be compared unless the variables included in the calculation are specified. It would be far more objective and comprehensive if an authorized institution made an informal decision based on formal assessments of the effectiveness of health-care services evaluated by health-care providers, of the value assessed by consumers, of efficacy described by biostatisticians, and of costs calculated by economists

Magnitude and Complexity of Rectal Mucosa HIV-1-Specific CD8+ T-Cell Responses during Chronic Infection Reflect Clinical Status

The intestinal mucosa displays robust virus replication and pronounced CD4+ T-cell loss during acute human immunodeficiency virus type 1 (HIV-1) infection. The ability of HIV-specific CD8+ T-cells to modulate disease course has prompted intensive study, yet the significance of virus-specific CD8+ T-cells in mucosal sites remains unclear.We evaluated five distinct effector functions of HIVgag-specific CD8+ T-cells in rectal mucosa and blood, individually and in combination, in relationship to clinical status and antiretroviral therapy (ART). In subjects not on ART, the percentage of rectal Gag-specific CD8+ T-cells capable of 3, 4 or 5 simultaneous effector functions was significantly related to blood CD4 count and inversely related to plasma viral load (PVL) (p<0.05). Polyfunctional rectal CD8+ T-cells expressed higher levels of MIP-1beta and CD107a on a per cell basis than mono- or bifunctional cells. The production of TNFalpha, IFN-gamma, and CD107a by Gag-specific rectal CD8+ T-cells each correlated inversely (p<0.05) with PVL, and MIP-1beta expression revealed a similar trend. CD107a and IFN-gamma production were positively related to blood CD4 count (p<0.05), with MIP-1beta showing a similar trend. IL-2 production by rectal CD8+ T-cells was highly variable and generally low, and showed no relationship to viral load or blood CD4 count.The polyfunctionality of rectal Gag-specific CD8+ T-cells appears to be related to blood CD4 count and inversely related to PVL. The extent to which these associations reflect causality remains to be determined; nevertheless, our data suggest a potentially important role for mucosal T-cells in limiting virus replication during chronic infection

Modulation of the immune response to Mycobacterium tuberculosis during malaria/M. tuberculosis co-infection

Tuberculosis (TB) causes significant morbidity and mortality on a global scale. The African region has 24% of the world's TB cases. TB overlaps with other infectious diseases such as malaria and HIV, which are also highly prevalent in the African region. TB is a leading cause of death among HIV-positive patients and co-infection with HIV and TB has been described as a syndemic. In view of the overlapping epidemiology of these diseases, it is important to understand the dynamics of the immune response to TB in the context of co-infection. We investigated the cytokine response to purified protein derivative (PPD) in peripheral blood mononuclear cells from TB patients co-infected with HIV or malaria and compared it to that of malaria- and HIV-free TB patients. A total of 231 subjects were recruited for this study and classified into six groups; untreated TB-positive, TB positive subjects on TB drugs, TB- and HIV-positive, TB- and malaria-positive, latent TB and apparently healthy control subjects. Our results demonstrate maintenance of interferon (IFN)-γ production in HIV and malaria co-infected TB patients in spite of lower CD4 counts in the HIV-infected cohort. Malaria co-infection caused an increase in the production of the T helper type 2 (Th2)-associated cytokine interleukin (IL)-4 and the anti-inflammatory cytokine IL-10 in PPD-stimulated cultures. These results suggest that malaria co-infection diverts immune response against M. tuberculosis towards a Th-2/anti-inflammatory response which might have important consequences for disease progression

A comparative phase I study of combination, homologous subtype-C DNA, MVA, and Env gp140 protein/adjuvant HIV vaccines in two immunization regimes

There remains an urgent need for a prophylactic HIV vaccine. We compared combined MVA and adjuvanted gp140 to sequential MVA/gp140 after DNA priming. We expected Env-specific CD4+ T-cells after DNA and MVA priming, and Env-binding antibodies in 100% individuals after boosting with gp140 and that combined vaccines would not compromise safety and might augment immunogenicity. Forty volunteers were primed three times with DNA plasmids encoding (CN54) env and (ZM96) gag-pol-nef at 0, 4 and 8 weeks then boosted with MVA-C (CN54 env and gag-pol-nef) and glucopyranosyl lipid adjuvant-aqueous formulation (GLA-AF) adjuvanted CN54gp140. They were randomised to receive them in combination at the same visit at 16 and 20 weeks (accelerated) or sequentially with MVA-C at 16, 20, and GLA-AF/gp140 at 24 and 28 weeks (standard). All vaccinations were intramuscular. Primary outcomes included ≥grade 3 safety events and the titer of CN54gp140-specific binding IgG. Other outcomes included neutralization, binding antibody specificity and T-cell responses. Two participants experienced asymptomatic ≥grade 3 transaminitis leading to discontinuation of vaccinations, and three had grade 3 solicited local or systemic reactions. A total of 100% made anti-CN54gp140 IgG and combining vaccines did not significantly alter the response; geometric mean titer 6424 (accelerated) and 6578 (standard); neutralization of MW965.2 Tier 1 pseudovirus was superior in the standard group (82 versus 45% responders,  = 0.04). T-cell ELISpot responses were CD4+ and Env-dominant; 85 and 82% responding in the accelerated and standard groups, respectively. Vaccine-induced IgG responses targeted multiple regions within gp120 with the V3 region most immunodominant and no differences between groups detected. Combining MVA and gp140 vaccines did not result in increased adverse events and did not significantly impact upon the titer of Env-specific binding antibodies, which were seen in 100% individuals. The approach did however affect other immune responses; neutralizing antibody responses, seen only to Tier 1 pseudoviruses, were poorer when the vaccines were combined and while T-cell responses were seen in >80% individuals in both groups and similarly CD4 and Env dominant, their breadth/polyfunctionality tended to be lower when the vaccines were combined, suggesting attenuation of immunogenicity and cautioning against this accelerated regimen

Louisville Seamount Trail: implications for geodynamic mantle flow models and the geochemical evolution of primary hotspots

The Louisville Seamount Trail is a 4300 km long volcanic chain that has been built in the past 80 m.y. as the Pacific plate moved over a persistent mantle melting anomaly or hotspot. Because of its linear morphology and its long-lived age-progressive volcanism, Louisville is the South Pacific counterpart of the much better studied Hawaiian-Emperor Seamount Trail. Together, Louisville and Hawaii are textbook examples of two primary hotspots that have been keystones in deciphering the motion of the Pacific plate relative to a set of "fixed" deep-mantle plumes. However, drilling during Ocean Drilling Program (ODP) Leg 197 in the Emperor Seamounts documented a large ~15° southward motion of the Hawaiian hotspot prior to 50 Ma. Is it possible that the Hawaiian and Louisville hotspots moved in concert and thus constitute a moving reference frame for modeling plate motion in the Pacific? Alternatively, could they have moved independently, as predicted by mantle flow models that reproduce the observed latitudinal motion for Hawaii but that predict a largely longitudinal shift for the Louisville hotspot? These two end-member geodynamic models were tested during Integrated Ocean Drilling Program (IODP) Expedition 330 to the Louisville Seamount Trail. In addition, existing data from dredged lavas suggest that the mantle plume source of the Louisville hotspot has been remarkably homogeneous for as long as 80 m.y. These lavas are predominantly alkali basalts and likely represent a mostly alkalic shield-building stage, which is in sharp contrast to the massive tholeiitic shield-building stage of Hawaiian volcanoes. Geochemical and isotopic data for the recovered lavas during Expedition 330 will provide insights into the magmatic evolution and melting processes of individual Louisville volcanoes, their progression from shield-building to postshield and (maybe) posterosional stages, the temperature and depth of partial melting of their mantle plume source, and the enigmatic long-lived and apparent geochemical homogeneity of the Louisville mantle source. Collectively, this will enable us to characterize the Louisville Seamount Trail as a product of one of the few global primary hotspots, to better constrain its plume-lithosphere interactions, and to further test the hypothesis that the Ontong Java Plateau formed from the plume head of the Louisville mantle plume around 120 Ma. During Expedition 330 we replicated the drilling strategy of Leg 197, the first expedition to provide compelling evidence for the motion of the Hawaiian mantle plume between 80 and 50 Ma. For that reason we targeted Louisville seamounts that have ages similar to Detroit, Suiko, Nintoku, and Koko Seamounts in the Emperor Seamount Trail. In total, five seamounts were drilled in the Louisville Seamount Trail: Canopus, Rigil, Burton, Achernar, and Hadar Guyots (old to young). By analyzing a large number of time-independent in situ lava flows (and other volcanic eruptive products) from these seamounts using modern paleomagnetic, 40Ar/39Ar geochronological, and geochemical techniques, we will be able to directly compare the paleolatitude estimates and geochemical signatures between the two longest-lived hotspot systems in the Pacific Ocean. We drilled into the summits of the five Louisville guyots and reached volcanic basement at four of these drilling targets. In two cases we targeted larger seamount structures and drilled near the flanks of these ancient volcanoes, and in the other three cases we selected smaller edifices that we drilled closer to their centers. Drilling and logging plans for each of these sites were similar, with coring reaching 522.0 meters below seafloor (mbsf) for Site U1374 and 232.9, 65.7, 11.5, 182.8, and 53.3 mbsf for Sites U1372, U1373, U1375, U1376, and U1377, respectively. Some Expedition 330 drill sites were capped with only a thin layer of pelagic ooze between 6.6 and 13.5 m thick, and, if present, these were cored by using a low-rotation gravity-push technique with the rotary core barrel to maximize recovery. However, at Sites U1373 and U1376 no pelagic ooze was present, and the holes needed to be started directly into cobble-rich hardgrounds. In all cases, the bulk of the seamount sediment cover comprised sequences of volcanic sandstones and various kinds of basalt breccia or basalt conglomerate, which often were interspersed with basaltic lava flows, the spatter/tephra products of submarine eruptions, or other volcanic products, including auto-brecciated flows or peperites. Also several intervals of carbonate were cored, with the special occurrence of a ~15 m thick algal limestone reef at Site U1376 on Burton Guyot. In addition, some condensed pelagic limestone units were recovered on three of the other seamounts, but these did not exceed 30 cm in thickness. Despite their limited presence in the drilled sediment, these limestones provide valuable insights for the paleoclimate record at high ~50° southern latitudes since Mesozoic times. Several Louisville sites progressed from subaerial conditions in the top of volcanic basement into submarine eruptive environments, or drilling of the igneous basement immediately started in submarine volcanic sequences, as was the case for Sites U1376 and U1377 on Burton and Hadar Guyots. At three sites we cored >100 m into the igneous basement: 187.3 m at Site U1372, 505.3 m at Site U1374, and 140.9 m at Site U1376. At the other sites we did not core into basement (Site U1375) or we cored only 38.2 m (Site U1377) because of unstable hole conditions. Even so, drilling during Expedition 330 resulted in a large number of in situ lava flows, pillow basalts, or other types of volcanic products such as auto-brecciated lava flows, intrusive sheets or dikes, and peperites. In particular, the three holes on Canopus and Rigil Guyots (the two oldest seamounts drilled in the Louisville Seamount Trail), resulted in adequate numbers of in situ lava flows to average out paleosecular variation, with probable eruption ages estimated at ~78 and 73 Ma, respectively. Remarkably, at all drill sites large quantities of hyaloclastites, volcanic sandstones, and basaltic breccias were also recovered, which in many cases show consistent paleomagnetic inclinations compared to the lava flows bracketing these units. For Site U1374 on Rigil Guyot we also observed a magnetic polarity reversal in the cored sequence. Overall, this is very promising for determining a reliable paleolatitude record for the Louisville Seamounts following detailed postcruise examinations. The deeper penetrations of several hundred meters required bit changes and reentries using free-fall funnels. Basement penetration rates were 1.8–2.5 m/h depending on drill depth. In total, 1114 m of sediment and igneous basement at five seamounts was drilled, and 806 m was recovered (average recovery = 72.4%). At Site U1374 on Rigil Guyot, a total of 522 m was drilled, with a record-breaking 87.8% recovery. Most outstandingly, nearly all Expedition 330 core material is characterized by low degrees of alteration, providing us with a large quantity of samples of mostly well-preserved basalt, containing, for example, pristine olivine crystals with melt inclusions, fresh volcanic glass, unaltered plagioclase, carbonate, zeolite and celadonite alteration minerals, various micro- and macrofossils, and, in one case, mantle xenoliths and xenocrysts. The large quantity and excellent quality of the recovered sample material allow us to address all the scientific objectives of this expedition and beyond