Population dynamics of Nyssomyia whitmani (Diptera: Psychodidae) in domestic and peridomestic environments in Northeast Argentina, a tegumentary leishmaniasis outbreak area.

In the present study, the temporal dynamics of the main vector of Leishmania braziliensis, Nyssomyia whitmani, was measured by monthly captures of phlebotominae sandflies during 5 consecutive years (from 2011 to 2016) in the Paranaense region of South America. The captures were performed in environments where the human-vector contact risk is high: domiciliary and peridomiciliary environments in a rural area endemic of tegumentary leishmaniasis. Nyssomyia whitmani was recorded as the dominant species of the phlebotominae ensemble in all domiciliary and peridomiciliary environments (House, Chicken Shed, Pigsty, and Forest Edge). Using generalized additive models, intra- and interannual fluctuations were observed, modulated by meteorological variables such as the minimum temperature and the accumulated precipitation 1 wk prior to capture. The installation of a pigsty by the farmer during the study period allowed us to observe and describe the so-called "pigsty effect" where the Ny. whitmani population was spatially redistributed, turning the pigsty as the environment that obtained the highest phlebotominae record counts, thus maintaining the farm overall abundance, supporting the idea that the environmental management of the peridomicile could have an impact on the reduction of epidemiological risk by altering the spatial distribution of the phlebotominae ensemble in the environments.

An Update of Cutaneous Melanoma Patients Treated in Adjuvancy With the Allogeneic Melanoma Vaccine VACCIMEL and Presentation of a Selected Case Report With In-Transit Metastases.

The CSF-470 vaccine (VACCIMEL) plus BCG and GM-CSF as adjuvants has been assayed in cutaneous melanoma patients. In the adjuvant randomized Phase II study CASVAC-0401, vaccinated patients had longer distant metastasis-free survival (DMFS) than those treated with IFNα2b. Five years after locking the data, an actualization was performed. The benefit in DMFS was maintained in the vaccinated group versus the IFNα2b-treated group (p = 0.035), with a median DMFS of 96 months for VACCIMEL and 13 months for IFNα2b. The favorable risk-benefit ratio was maintained. DMFS was also analyzed as a single cohort in all the IIB, IIC, and III patients (n = 30) who had been treated with VACCIMEL. The median DMFS was 169 months, and at 48 months follow-up, it was 71.4%, which was not statistically different from DMFS of previously published results obtained in adjuvancy with ipilimumab, pembrolizumab, nivolumab, or dabrafenib/trametinib. The possible toxicity of combining VACCIMEL with anti-immune checkpoint inhibitors (ICKi) was analyzed, especially since VACCIMEL was co-adjuvated with BCG in every vaccination. A patient with in-transit metastases was studied to produce a proof of concept. During treatment with VACCIMEL, the patient developed T-cell clones reactive towards tumor-associated antigens. Three years after ending the VACCIMEL study, the patient progressed and was treated with ICKi. During ICKi treatment, the patient did not reveal any toxicity due to previous BCG treatment. When she recurred after a 4-year treatment with nivolumab, a biopsy was obtained and immunohistochemistry and RNA-seq were performed. The tumor maintained expression of tumor-associated antigens and HLA-I and immune infiltration, with immunoreactive and immunosuppressive features. VACCIMEL plus BCG and GM-CSF is an effective treatment in adjuvancy for stages IIB, IIC, and III cutaneous melanoma patients, and it is compatible with subsequent treatments with ICKi.

Peripheral changes in immune cell populations and soluble mediators after anti-PD-1 therapy in non-small cell lung cancer and renal cell carcinoma patients.

Patients with non-small cell lung cancer (NSCLC) and renal cell carcinoma (RCC) have shown benefit from anti-PD-1 therapies. However, not all patients experience tumor shrinkage, durable responses or prolonged survival, demonstrating the need to find response markers. In blood samples from NSCLC and RCC patients obtained before and after anti-PD-1 treatment, we studied leukocytes by complete blood cell count, lymphocyte subsets using flow cytometry and plasma concentration of nine soluble mediators, in order to find predictive biomarkers of response and to study changes produced after anti-PD-1 therapy. In baseline samples, discriminant analysis revealed a combination of four variables that helped differentiate stable disease-response (SD-R) from progressive disease (PD) patients: augmented frequency of central memory CD4+ T cells and leukocyte count was associated with response while increased percentage of PD-L1+ natural killer cells and naïve CD4+ T cells was associated with lack of response. After therapy, differential changes between responders and non-responders were found in leukocytes, T cells and TIM-3+ T cells. Patients with progressive disease showed an increase in the frequency of TIM-3 expressing CD4+ and CD8+ T cells, whereas SD-R patients showed a decrease in these subsets. Our findings indicate that a combination of immune variables from peripheral blood (PB) could be useful to distinguish response groups in NSCLC and RCC patients treated with anti-PD-1 therapy. Frequency of TIM-3+ T cells showed differential changes after treatment in PD vs SD-R patients, suggesting that it may be an interesting marker for monitoring progression during therapy.

Hantavirus Pulmonary Syndrome Risk in Entre Ríos, Argentina.

Hantavirus pulmonary syndrome (HPS) is a severe emerging endemic disease of the Americas. Because hantavirus reservoirs are sylvatic rodents, HPS risk has been associated with occupational and recreational activities in natural and rural environments. The aim of this study was to analyze the risk of HPS in an endemic province of Argentina. For this, we explored the relationship between HPS cases occurring in Entre Ríos province between 2004 and 2015 and climate, vegetation, landscape, reservoir population, and rodent community characteristics by means of generalized linear models. We modeled HPS occurrence at each site, and both the incidence and number of cases grouped by department. The resulting best model of each analysis was applied in a GIS to build HPS risk maps. Risk of occurrence of HPS increased with tree cover and decreased with distance to rivers. We identified the south of Entre Ríos as the area with higher HPS risk, and therefore, where HPS prevention measures should be more urgently applied. Risk maps based on data available in the public domain are a useful tool that should be used by decision makers to concentrate surveillance and control efforts in those areas with highest HPS risk.

Dissecting the Immune Stimulation Promoted by CSF-470 Vaccine Plus Adjuvants in Cutaneous Melanoma Patients: Long Term Antitumor Immunity and Short Term Release of Acute Inflammatory Reactants.

As cutaneous melanoma (CM) currently remains with a bleak prognosis, thorough investigation of new treatment options are of utmost relevance. In the phase II/III randomized clinical trial (CASVAC-0401), the repeated immunization of stages IIB-III CM patients with the irradiated, allogeneic cellular CSF-470 vaccine plus the adjuvants bacillus Calmette-Guerin (BCG) and recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) demonstrated a significant benefit over IFN-alpha2B treatment in distant metastasis-free survival. Here we present on the short and long term immune monitoring results after completing the 2-year protocol; a continuation of the previous report by Mordoh et al. (1). We demonstrate that the repeated CSF-470 vaccinations stimulated a long term cellular and humoral immunity response directed against the vaccine antigens. In the case of 2 patients, we are able to show that a similar immune response was generated against autologous antigens. Evaluation of inhibitory receptor co-expression on patient's T cells indicates that the vaccination protocol did not stimulate T cell exhaustion. In order to better understand the basis for the efficacious vaccine responses observed, we investigated the short term immune events following vaccine injection. A significant increase in C-reactive protein (CRP) and IL-6 was observed 24 h after vaccination, with in vitro studies suggesting IL-6 production occurs in the vaccine site. We demonstrate that CRP enhances the cytotoxicity of peripheral blood mononuclear cells (PBMC) against melanoma cells in an in vitro model. Additionally, CRP stimulates the release of pro and anti-inflammatory cytokines from PBMC. As our results demonstrate that successive vaccinations with CSF-470 plus adjuvants promoted an increase in both anti-tumor innate and adaptive immunity, we propose a subsequent model of action.

Long-Term Study of a Hantavirus Reservoir Population in an Urban Protected Area, Argentina.

Green spaces in urban areas can play a key role in protecting wildlife. However, the presence of wildlife in urban areas can lead to human health risks. Although the presence of the rodent species Oligoryzomys flavescens (hantavirus reservoir) has been recorded in cities of Argentina, its population dynamics in this type of habitat is still unknown. Here, we evaluated: (1) long-term spatial and temporal patterns of O. flavescens abundance and how these patterns were influenced by weather factors and (2) the seroprevalence of hantavirus and the identity of the viral lineage circulating in the population that inhabits the Costanera Sur Ecological Reserve, a protected area in the city of Buenos Aires. Genetic results confirmed that the pathogenic ANDES Central Lechiguanas virus is present in O. flavescens populations inhabiting this urban reserve. Abundance of O. flavescens showed interannual and seasonal fluctuations, with maximum values in winter and spring and minimum ones in summer and autumn. Summers with the highest abundances of O. flavescens were preceded by warmer winters, while winters with lower abundances were preceded by warmer summers. On the other hand, accumulated precipitations in the previous 6 months positively affected winter abundance. These results could help the authorities in charge of the green spaces of Buenos Aires to identify priority areas and times of the year for the implementation of preventive measures that minimize the contact of rodents with visitors. Such measures could be intensified when winters are warmer than normal, and summers are cooler and wetter than normal.

Commensal Rodents in the City of Buenos Aires: A Temporal, Spatial, and Environmental Analysis at the Whole City Level.

Commensal rodents are some of the main vertebrate pests in urban areas; however, knowledge about factors that favor them at large scales is scarce. We studied spatial and temporal variations in rodent infestation levels at the whole city scale using the complaints of rodent sightings and questionnaire surveys to city neighbors. Demographic, socio-economic, and environmental characteristics handled with a geographic information system were evaluated as possible indicators of rodent infestation. The number of rodent sightings was lower in months with low mean temperature with two months time lag and higher in areas with high number of meat and metal industries. Rodent infestation estimated by the questionnaire survey showed spatial autocorrelation defining large areas with similar infestation levels. It decreased when the apartment density increased, while increased when the proportion of area occupied by shantytowns, the density of meat industries, and the proportion of area occupied by moderate urban development increased. Rodent control programs at the whole city level would have better results if public health pest agencies and/or governments will focus the efforts on areas with more precarious conditions as well as the industrial areas in the cold season when have lower rodent abundances.

Partial genetic characterization of Seoul hantavirus in rats from Buenos Aires City, Argentina, and generation of a Seoul recombinant nucleoprotein antigen

Hemorrhagic fever with renal syndrome (HFRS) is a severe infectious disease characterized by fever, hemorrhage, renal impairment, and thrombocytopenia. At least seven hantaviruses cause HFRS: Hantaan, Seoul (SEOV) (distributed worldwide), Dobrava/Belgrade, Saaremaa, Amur, Thailand and Puumala. To investigate the epidemiology of HFRS and virus transmission in Argentina, we constructed a prokaryotic plasmid encoding the SEOV rNP, of 430 amino acids. After expression, the rNP was tested as an antigen for use in an enzyme-linked immunosorbent assay for infection diagnosis. To determine the current level of virus transmission in wild brown rats or Norway rats (Rattus norvegicus) captured in Buenos Aires City, Argentina, we tested tissues from rats that were determined to be serologically positive for the SEOV, and the viral genome were detected by RT-PCR using specific primers for two fragments of M segment-encoding Gn and Gc proteins. The viral genome was detected in 11 of 21 seropositive rats (52.4 per cent) captured in two parklands. Sequence analysis of a 333-nt region of the Gc-encoding M segment revealed 97 per cent and 96 per cent identity with strains of SEOV from Baltimore and Brazil, respectively. Our genetic data confirm a very low diversity among SEOV virus strains...

A febre hemorrágica com síndrome renal (FHSR) é uma doença grave, caracterizada por febre, hemorragia, falência renal e trombocitopenia. Pelo menos sete hantavírus causam a FHSR: Hantaan, Seoul (SEOV) (de distribuição global), Dobrava-Belgrade, Saaremaa, Amur, Thailand e Puumala. Para investigar a epidemiologia da FHRS e a transmissão viral na Argentina, criamos um plasmídio procariótico que "codifica" a nucleoproteína recombinante do vírus SEOV de 430 aminoácidos. Após a expressão, a nucleoproteína recombinante foi testada como antígeno para uso em ensaio imunoenzimático (ELISA) para diagnóstico da infecção. Para determinar o nível atual de transmissão viral em populações de ratos-marrons ou ratazanas (Rattus norvegicus) capturadas na cidade de Buenos Aires, Argentina, testamos tecidos de ratos selecionados para serem sorologicamente positivos para o vírus SEOV, e o seu genoma viral foi detectado após submetido a RT-PCR utilizando primers específicos para dois fragmentos de proteínas Gn e Gc codificadas pelo segmento M. O genoma viral foi detectado em 11 das 21 ratazanas soropositivas (52,4 por cento), previamente capturadas em dois parques. A análise sequencial de uma região gênica (333 nt) do segmento M "codificador" da proteína Gc apresentou 97 por cento e 96 por cento de similaridade com as cepas de SEOV coletadas em Baltimore e no Brasil, respectivamente. Os dados genéticos levantados confirmam a informação de que há uma diversidade muito pequena entre as cepas do vírus SEOV...

Prevalence of hantavirus infection in wild Rattus norvegicus and R. rattus populations of Buenos Aires City, Argentina.

OBJECTIVE: To determine the prevalence of infection with hantaviruses in wild Rattus norvegicus and R. rattus populations in areas of Buenos Aires city. METHODS: Norway rats and black rats were trapped from 2003 to 2005 at 10 sites throughout Buenos Aires city. Blood samples for serological analysis were collected by cardiac puncture from all individuals captured and tested by enzyme-linked immunosorbent assay for Seoul hantavirus- and Andes virus-specific IgG antibodies. RESULTS: Hantavirus seroprevalence was 11.9% in R. norvegicus (n = 151), varying between 0% and 26.1% depending on the site. Bigger sexually active males were more likely to be infected with Seoul virus than females or juvenile individuals. No antibody-reactive rodents were detected among 33 R. rattus analysed. CONCLUSION: Hantavirus infections are geographically widespread in Buenos Aires city and confirm that they have been enzootic there for at least 20 years.