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i.v. injected Abs have demonstrated protection against simian HIV infection in rhesus macaques, paving the way for the Antibody Mediated Prevention trial in which at-risk individuals for HIV received an i.v. infusion of the HIV broadly neutralizing Ab VRC01. However, the time needed for these Abs to fully distribute and elicit protection at mucosal sites is still unknown. In this study, we interrogate how long it takes for Abs to achieve peak anatomical levels at the vaginal surface following i.v. injection. Fluorescently labeled VRC01 and/or Gamunex-C were i.v. injected into 24 female rhesus macaques (Macaca mulatta) with vaginal tissues and plasma acquired up to 2 wk postinjection. We found that Ab delivery to the vaginal mucosa occurs in two phases. The first phase involves delivery to the submucosa, occurring within 24 h and persisting beyond 1 wk. The second phase is the delivery through the stratified squamous epithelium, needing ∼1 wk to saturate the stratum corneum. This study has important implications for the efficacy of immunoprophylaxis targeting pathogens at the mucosa.

Messenger RNA has now been used to vaccinate millions of people. However, the diversity of pulmonary pathologies, including infections, genetic disorders, asthma and others, reveals the lung as an important organ to directly target for future RNA therapeutics and preventatives. Here we report the screening of 166 polymeric nanoparticle formulations for functional delivery to the lungs, obtained from a combinatorial synthesis approach combined with a low-dead-volume nose-only inhalation system for mice. We identify P76, a poly-β-amino-thio-ester polymer, that exhibits increased expression over formulations lacking the thiol component, delivery to different animal species with varying RNA cargos and low toxicity. P76 allows for dose sparing when delivering an mRNA-expressed Cas13a-mediated treatment in a SARS-CoV-2 challenge model, resulting in similar efficacy to a 20-fold higher dose of a neutralizing antibody. Overall, the combinatorial synthesis approach allowed for the discovery of promising polymeric formulations for future RNA pharmaceutical development for the lungs.

Human immunodeficiency virus (HIV) vaccines have not been successful in clinical trials. Dimeric IgA (dIgA) in the form of secretory IgA is the most abundant antibody class in mucosal tissues, making dIgA a prime candidate for potential HIV vaccines. We coupled Positron Emission Tomography (PET) imaging and fluorescent microscopy of 64Cu-labeled, photoactivatable-GFP HIV (PA-GFP-BaL) and fluorescently labeled dIgA to determine how dIgA antibodies influence virus interaction with mucosal barriers and viral penetration in colorectal tissue. Our results show that HIV virions rapidly disseminate throughout the colon two hours after exposure. The presence of dIgA resulted in an increase in virions and penetration depth in the transverse colon. Moreover, virions were found in the mesenteric lymph nodes two hours after viral exposure, and the presence of dIgA led to an increase in virions in mesenteric lymph nodes. Taken together, these technologies enable in vivo and in situ visualization of antibody-virus interactions and detailed investigations of early events in HIV infection., Author summary Vaccines provide protection in humans by eliciting the production of antibodies when exposed to a specific pathogen. Currently, an effective human immunodeficiency virus (HIV) vaccine does not exist. Since the beginning of the HIV epidemic, approximately 38 million people have died, creating the need to develop an HIV vaccine. The most common antibody in the organs that are exposed to HIV is dimeric IgA (dIgA). Here, we used multiple imaging techniques to determine how HIV travels throughout the colon once introduced into the body and how dIgA influences HIV movement in the rectum. We found that dIgA increased the amount of HIV found in the colon, the distance it travelled, and the depth into tissues that HIV penetrated. dIgA also increased the amount of HIV in the mesenteric lymph nodes two hours after viral exposure. Our study shows these imaging technologies can be used to examine interactions between viruses and antibodies in early HIV infection in the natural context of the anatomy and physiology of the rhesus macaque model.

In Sub-Saharan Africa, young women 15–24 years of age account for nearly 30% of all new HIV infections, however, biological and epidemiological factors underlying this disproportionate infection rate are unclear. In this study, we assessed biological contributors of SIV/HIV susceptibility in the female genital tract (FGT) using adolescent (n = 9) and adult (n = 10) pigtail macaques (PTMs) with weekly low-dose intravaginal challenges of SIV. Immunological variables were captured in vaginal tissue of PTMs by flow cytometry and cytokine assays. Vaginal biopsies were profiled by proteomic analysis. The vaginal microbiome was assessed by 16S rRNA sequencing. We were powered to detect a 2.2-fold increase in infection rates between age groups, however, we identified no significant differences in susceptibility. This model cannot capture epidemiological factors or may not best represent biological differences of HIV susceptibility. No immune cell subsets measured were significantly different between groups. Inflammatory marker MCP-1 was significantly higher (adj p = .02), and sCD40L trended higher (adj p = .06) in vaginal cytobrushes of adults. Proteomic analysis of vaginal biopsies showed no significant (adj p

In Sub-Saharan Africa, young women 15–24 years of age account for nearly 30% of all new HIV infections, however, biological and epidemiological factors underlying this disproportionate infection ra...

There is a clear need for low-cost, self-applied, long-lasting approaches to prevent human immunodeficiency virus (HIV) infection in both men and women, even with the advent of pre-exposure prophylaxis (PrEP). Broadly neutralizing antibodies represent an option to improve HIV prophylaxis, but intravenous delivery, cold-chain stability requirements, low cervicovaginal concentrations, and cost may preclude their use. Here, we present an approach to express the anti-GP120 broadly neutralizing antibody PGT121 in the primary site of inoculation, the female reproductive tract, using synthetic mRNA. Expression is achieved through aerosol delivery of unformulated mRNA in water. We demonstrated high levels of antibody expression for over 28 days with a single mRNA administration in the reproductive tract of sheep. In rhesus macaques, neutralizing antibody titers in secretions developed within 4 h and simian-HIV (SHIV) infection of ex vivo explants was prevented. Persistence of PGT121 in vaginal secretions and epithelium was achieved through the incorporation of a glycosylphosphatidylinositol (GPI) anchor into the heavy chain of the antibody. Overall, we present a new paradigm to deliver neutralizing antibodies to the female reproductive tract for the prevention of HIV infections., Graphical Abstract, Santangelo and colleagues report on the aerosol delivery of synthetic mRNAs, diluted in water, to the cervicovaginal epithelium, which expresses membrane-anchored broadly neutralizing antibodies against HIV. In two large animal models, anchored PGT121 was widely dispersed in the reproductive tract and was protective against a SHIV explant challenge.

Aim: While the therapeutic potential for current long-acting (LA) antiretroviral therapy (ART) is undeniable, ligand-decorated nanoformulated LA-ART could optimize drug delivery to viral reservoirs. The development of decorated ART hinges, however, on formulation processes and manufacture efficiencies. To this end, we compared manufacture and purification techniques for ligand-decorated antiretroviral drug nanocrystals. Materials & methods: Ligand-decorated nanoparticle manufacturing was tested using folic acid (FA) nanoformulated cabotegravir. Results: Direct manufacturing of FA-cabotegravir resulted in stable particles with high drug loading and monocyte–macrophage targeting. A one step ‘direct’ scheme proved superior over differential centrifugation or tangential flow filtration facilitating particle stability and preparation simplicity and efficiency. Conclusion: Direct manufacturing of FA nanoparticles provides a path toward large-scale clinical grade manufacturing of cell-targeted LA-ART., Lay abstract Folic acid (FA) decoration on the surface of nanocrystals can be achieved by mixing FA conjugated poloxamer 407 (FA-P407) and native P407 in varied ratios followed by size reduction by homogenization and differential centrifugation or tangential flow filtration to remove excess unbound polymers. The optimized manufacturing scheme is by direct homogenization with predetermined quantity of FA conjugated P407. Direct manufacturing method yields stable homogenous nanoparticles with high drug loading.

Vertical transmission of human immunodeficiency virus (HIV) can occur in utero, during delivery, and through breastfeeding. We utilized Positron Emission Tomography (PET) imaging coupled with fluorescent microscopy of 64Cu-labeled photoactivatable-GFP-HIV (PA-GFP-BaL) to determine how HIV virions distribute and localize in neonatal rhesus macaques two and four hours after oral viral challenge. Our results show that by four hours after oral viral exposure, HIV virions localize to and penetrate the rectal mucosa. We also used a dual viral challenge with a non-replicative viral vector and a replication competent SHIV-1157ipd3N4 to examine viral transduction and dissemination at 96 hours. Our data show that while SHIV-1157ipd3N4 infection can be found in the oral cavity and upper gastrointestinal (GI) tract, the small and large intestine contained the largest number of infected cells. Moreover, we found that T cells were the biggest population of infected immune cells. Thus, thanks to these novel technologies, we are able to visualize and delineate of viral distribution and infection throughout the entire neonatal GI tract during acute viral infection., Author summary Approximately 1.8 million children are currently living with human immunodeficiency virus (HIV). While mother-to-child HIV transmission can occur in utero and during delivery, it most commonly occurs through breastfeeding, creating the need to understand how the virus moves throughout the body and infects the infant once breast milk is consumed. Here, we used multiple imaging techniques and PCR to determine how HIV distributes throughout the gastrointestinal tract after oral viral exposure and in which tissues and cell types become acutely infected. We found that HIV rapidly spreads throughout and penetrates the entire gastrointestinal tract as early as four hours after exposure. We also found that the intestine contained the largest number of infected cells at 96 hours and that most cells infected were T cells. Our study shows that these imaging technologies allow for the examination of viral distribution and infection in a rhesus macaque model.

These authors contributed equally : Kevin E. Lindsay, Sushma M. Bhosle and Chiara Zurla.; International audience; Visualization of the spatio-temporal trafficking of vaccines after their delivery would help evaluate the efficacy of candidate formulations and aid their rational design for preclinical and translational studies. Here, we show that a dual radionuclide-near-infrared probe allows for quantitative, longitudinal and non-invasive monitoring, via positron emission tomography-computed tomography and near-infrared imaging of cynomolgus macaques, of the trafficking dynamics to draining lymph nodes of a model messenger RNA vaccine labelled with the probe. After intramuscular administration of the vaccine to the monkeys, we observed the dynamics of the mRNA vaccine at the injection site and in the draining lymph nodes, performed cellular analyses of the involved tissues using flow cytometry and identified through immunofluorescence that professional antigen-presenting cells are the primary cells containing the injected mRNA and encoding the antigen. This approach may reveal spatio-temporal determinants of vaccine efficacy in preclinical and translational studies employing large mammals.

Supplemental Digital Content is Available in the Text., Background: Antiretroviral drug discovery and formulation design will facilitate viral clearance in infectious reservoirs. Although progress has been realized for selected hydrophobic integrase and nonnucleoside reverse transcriptase inhibitors, limited success has been seen to date with hydrophilic nucleosides. To overcome these limitations, hydrophobic long-acting drug nanoparticles were created for the commonly used nucleoside reverse transcriptase inhibitor, lamivudine (2′,3′-dideoxy-3′-thiacytidine, 3TC). Methods: A 2-step synthesis created a slow-release long-acting hydrophobic 3TC. Conjugation of 3TC to a fatty acid created a myristoylated prodrug which was encased into a folate-decorated poloxamer 407. Both in vitro antiretroviral efficacy in human monocyte-derived macrophages and pharmacokinetic profiles in mice were evaluated for the decorated nanoformulated drug. Results: A stable drug formulation was produced by poloxamer encasement that improved monocyte–macrophage uptake, antiretroviral activities, and drug pharmacokinetic profiles over native drug formulations. Conclusions: Sustained release of long-acting antiretroviral therapy is a new therapeutic frontier for HIV/AIDS. 3TC depot formation in monocyte-derived macrophages can be facilitated through stable subcellular internalization and slow drug release.

Humanized mice have emerged as a testing platform for HIV-1 pathobiology by reflecting natural human disease processes. Their use to study HIV-1 biology, virology, immunology, pathogenesis and therapeutic development has served as a robust alternative to more-well developed animal models for HIV/AIDS. A critical component in reflecting such human pathobiology rests in defining the tissue and cellular sites for HIV-1 infection. To this end, we examined the tissue sites for viral infection in bone marrow, blood, spleens, liver, gut, brain, kidney and lungs of human CD34+ hematopoietic stem cell engrafted virus-infected NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ mice. Cells were analyzed by flow cytometry and sorted from species mixtures defined as CD34+ lineage negative progenitor cells, CD14+CD16+ monocyte-macrophages and central, stem cell and effector memory T cells. The cell distribution and viral life cycle were found dependent on the tissue compartment and time of infection. Cell subsets contained HIV-1 total and integrated DNA as well as multi-spliced and unspliced RNA in divergent proportions. The data support the idea that humanized mice can provide a means to examine the multifaceted sites of HIV-1 replication including, but not limited to progenitor cells and monocyte-macrophages previously possible only in macaques and human.

During studies to extend the half-life of crystalline nanoformulated antiretroviral therapy (nanoART) the mixed lineage kinase-3 inhibitor URMC-099, developed as an adjunctive neuroprotective agent was shown to facilitate antiviral responses. Long-acting ritonavir-boosted atazanavir (nanoATV/r) nanoformulations co-administered with URMC-099 reduced viral load and the numbers of HIV-1 infected CD4+ T-cells in lymphoid tissues more than either drug alone in infected humanized NOD/SCID/IL2Rγc−/− mice. The drug effects were associated with sustained ART depots. Proteomics analyses demonstrated that the antiretroviral responses were linked to affected phagolysosomal storage pathways leading to sequestration of nanoATV/r in Rab-associated recycling and late endosomes; sites associated with viral maturation. URMC-099 administered with nanoATV induced a dose-dependent reduction in HIV-1p24 and reverse transcriptase activity. This drug combination offers a unique chemical marriage for cell-based viral clearance.From the Clinical EditorAlthough successful in combating HIV-1 infection, the next improvement in antiretroviral therapy (nanoART) would be to devise long acting therapy, such as intra-cellular depots. In this report, the authors described the use of nanoformulated antiretroviral therapy given together with the mixed lineage kinase-3 inhibitor URMC-099, and showed that this combination not only prolonged drug half-life, but also had better efficacy. The findings are hoped to be translated into the clinical setting in the future.

Long acting nanoformulated antiretroviral therapy (nanoART) can sustain plasma drug levels and improve its biodistribution. Cell targeted-nanoART can achieve this and bring drug efficiently to viral reservoirs. However, if such improvements affect antiretroviral responses remains unknown. To these ends, we tested folic acid (FA)-linked poloxamer407 coated-ritonavir boosted atazanavir (FA-nanoATV/r) nanoparticles for their ability to affect chronic HIV-1 infection in humanized mice. Following three every other week 100 mg/kg FA-nanoATV/r intramuscular injection administered to infected animals viral RNA was at or below the detection limit, cell-associated HIV-1p24 reduced and CD4+ T cell counts protected. The dosing regimen improved treatment outcomes more than two fold from what was reported for untargeted nanoATV/r. We posit that these nanoformulations have potential for translation to human use.

Background Long-acting nanoformulated antiretroviral therapy (nanoART) is designed to improve patient regimen adherence, reduce systemic drug toxicities, and facilitate clearance of human immunodeficiency virus type one (HIV-1) infection. While nanoART establishes drug depots within recycling and late monocyte-macrophage endosomes, whether or not this provides a strategic advantage towards viral elimination has not been elucidated. Results We applied quantitative SWATH-MS proteomics and cell profiling to nanoparticle atazanavir (nanoATV)-treated and HIV-1 infected human monocyte-derived macrophages (MDM). Native ATV and uninfected cells served as controls. Both HIV-1 and nanoATV engaged endolysosomal trafficking for assembly and depot formation, respectively. Notably, the pathways were deregulated in opposing manners by the virus and the nanoATV, likely by viral clearance. Paired-sample z-scores, of the proteomic data sets, showed up- and down- regulation of Rab-linked endolysosomal proteins. NanoART and native ATV treated uninfected cells showed limited effects. The data was confirmed by Western blot. DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking. Conclusions We posit that modulation of endolysosomal pathways by antiretroviral nanoparticles provides a strategic path to combat HIV infection. Electronic supplementary material The online version of this article (doi:10.1186/s12977-014-0133-5) contains supplementary material, which is available to authorized users.

El objetivo del presente estudio fue determinar la frecuencia de anticuerpos contra el virus de la Parainfluenza 3 (VPI3) en porcinos beneficiados en la ciudad de Lima. Se colectaron muestras de sangre de cerdos de ambos sexos y de 2 a 6 meses de edad, provenientes de granjas tecnificadas (n=192) y de crianza de traspatio (n= 192) para la deteccion de anticuerpos contra el VPI3 mediante la tecnica de neutralizacion viral. El 5.5 ± 2.3% (21/384) de las muestras tuvieron anticuerpos contra el VPI3, donde el 5.2 ± 1.1% (10/192) y el 5.7 ± 1.2% (11/192) correspondio a animales de crianza tecnificada y de traspatio, respectivamente. Los titulos de anticuerpos tuvieron un rango de 4 a 64 y fueron detectados en animales mayores de 3 meses de edad. Los resultados indican la presencia del VPI3 en la poblacion de cerdos

El efecto de la GnRH sobre el porcentaje de supervivencia embrionaria durante el periodo del reconocimiento maternal de la preñez, fue evaluado en 67 alpacas adultas receptivas al macho y que a la ecografía presentaban un folículo preovulatorio ≥7 mm. Los animales se distribuyeron en 3 grupos: G0 (n=23) como control; G1(n=22) que recibió 6 µg GnRH el día 4 post-ovulación; y G2(n=22) que recibió 4 µg GnRH los días 8 y 9 postovulación. La ocurrencia de ovulación fue confirmada por la detección del cuerpo lúteo (CL) en el día 5 post-ovulación. La fertilización y viabilidad de la gestación fue monitoreada por el desarrollo del CL, cambios estructurales de los cuernos uterinos y presencia de la vesícula embrionaria en los días 12, 18, 25 y 30 post-ovulación por ecografía transrectal de los ovarios y útero. Simultáneamente se evaluó la receptividad sexual de la hembra al macho. El número de hembras que ovularon fue 20, 22 y 21 para G0, G1 y G2, respectivamente, considerándose que se encontraban gestantes (día 5); sin embargo, en el día 12 el porcentaje de supervivencia fue de 90.5% en G1, 75% en G0 y 76.2% en G2. El porcentaje de supervivencia embrionaria fue significativamente mayor en el día 25 y 30 (p, The effect of GnRH administered around the time of maternal recognition of prenancy on the embryonic survival rate was studied. It was used 67 adult female alpacas sexually receptive to the male, and bearing a preovulatory follicle ≥7mm, detected by rectal ultrasonography. Animals were distributed in three groups: G0 (n=23) as control; G1 (n=22) received 6 µg GnRH on day 4 after ovulation; and G2 (n=22) received 4 µg GnRH on days 8 and 9 after ovulation. The ocurrence of ovulation was confirmed through the detection of a corpus luteum (CL) on day 5 after ovulation. Fertilization and gestation viab-ility were monitored by CL development, structural changes of the uterine horns and presence of the embryonic vesicle on days 12, 18, 25 and 30 after ovulation, using rectal ultrasonography. The female sexual receptivity to the male was simultaneously evaluated. The number of females that ovulate was 20, 22 and 21 in G0, G1, and G2 respectively, and was presumed that conception has occurred. However, on day 12, the embryo survival rate was 90.5% in G1, 75.0% in G0,and 76.2% in G2. These values were statistically different (p

The aim of the present study was to determine the prevalence of Bovine Viral Diarrhea virus (BVDV) and Border Disease virus (BDV) in breeding sheep from a large cooperative farm in the central highlands of Peru. Blood samples from apparently healthy sheep of 4 years old, both sexes (female = 165; male = 165) were collected for antibodies detection against BVDV and BDV using the virus neutralisation test. The 2.1 ± 1.5% (7/330) and 28.5 ± 4.9% (94/330) of breeding sheep had antibodies against BVDV and BDV respectively, with antibodies titers of 1:2 and 1:16. There was significant association between sex and presence of antibodies against BDV (females: 53.3 ± 7.6%; males: 3.6 ± 2.9%) (p

El objetivo del estudio fue determinar la frecuencia de anticuerpos contra Leptospira spp en porcinos provenientes de crianza tecnificada (5 granjas, n=163) y de traspatio (11 criaderos, n=133) del valle de Lima. Se colectaron muestras de sangre de porcinos (n=296) durante el beneficio en dos mataderos de la ciudad de Lima, para la deteccion de anticuerpos contra ocho serovares de Leptospira mediante la prueba de microaglutinacion. El 85.8 ± 3.9% (254/296) de las muestras fue positivo a uno o mas serovares de Leptospira. El 89.6 ± 4.7% (146/163) y el 82.1 ± 6.5% (108/133) de las muestras de porcinos de crianza tecnificada y de traspatio, respectivamente, tuvieron anticuerpos contra Leptospira. Los serovares icterohaemorrhagiae, pomona y georgia fueron los mas frecuentes en ambos tipos de crianza. No se detectaron anticuerpos contra los serovares bratislava y grippothyphosa. Los titulos de anticuerpos tuvieron un rango entre 100 a 400 en ambos tipos de crianza, pero titulos de anticuerpos de 800 a 1600 fueron detectados con mayor frecuencia en animales de traspatio. La combinacion icterohaemorrhagiae y pomona fue la mas comun en ambos tipos de crianza. No hubo asociacion entre la presencia de anticuerpos contra Leptospira y la procedencia de los porcinos evaluados.

El objetivo del presente estudio fue determinar la prevalencia del virus de la diarrea viral bovina (VDVB) y de animales persistentemente infectados (PI) en bovinos productores de leche en la Irrigación de Majes, Arequipa. En la primera fase del estudio, se colectaron 204 muestras de leche de tanque (una por hato) en la planta local de procesamiento de leche, para la detección de anticuerpos contra el VDVB mediante la prueba de ELISA indirecta. En la segunda fase se seleccionaron de modo arbitrario a 57 hatos con anticuerpos contra el VDVB con densidades ópticas (DO) iguales o mayores a 0.900. Se colectaron 286 muestras de suero sanguíneo de estos hatos para conocer el estado serológico de cada animal y la búsqueda de animales PI mediante ELISA indirecta y ELISA de captura, respectivamente. En la tercera fase se obtuvieron muestras de suero de la totalidad de terneras y vaquillas (n = 20) de tres hatos que tuvieron animales PI a fin de buscar más animales PI. El 98.0 ± 1.9% (200/204) de las muestras de leche resultaron positivas a anticuerpos contra el VDVB con DO que fluctuaron entre 0.300 a 2.350. De las 286 muestras de suero pertenecientes a los 57 hatos, el 47.2% (135/286) tuvieron anticuerpos contra el VDVB; además, dentro del grupo de animales seronegativos se detectaron cuatro (2.7%) animales PI que pertenecieron a tres de los 57 hatos muestreados. En los animales de riesgo de los tres hatos (n = 20), se detectaron otros dos animales PI, sumando un total de seis (4.0%); además, estos seis animales PI no presentaron anticuerpos contra el VDVB. En conclusión, los resultados muestran una amplia distribución del VDVB en la población bovina de los hatos de la Irrigación Majes; así mismo, las muestras que presentan altas DO evidencian la presencia de animales PI y, por último, la muestra de leche de tanque reemplaza al suero en la detección de anticuerpos contra el VDVB, The objective of this study was to determine the prevalence of bovine viral diarrhoea virus (BVDV) and persistently infected (PI) animals in dairy herds located in Majes, Arequipa, Peru. In the first phase, 204 bulk tank milk samples from to 204 herds were collected al the local commercial processing plant in Majes for detection of antibodies against BVDV by indirect ELISA test. In the second phase, 286 blood samples were collected from 57 strong positive (Optical Density [OD] ? 0.900) herds for the detection of antibodies against BVDV and in PI animals by the indirect and capture ELISAs respectively. In the third phase, blood samples were collected from all animals ranging from 6 to 24 months of age from three herds that had at least one PI animal. The prevalence of BVDV in the 204 herds was 98.0 ± 1.9% (200/204), and the level of antibodies ranged from 0.300 to 2.350 OD. The 47.2% (135/286) of serum samples had antibodies against BVDV, and within the negative samples, four (2.7%) animals from three herds were PI. Later on, additional PI animals were also detected in these three herds, and none of the six (4.0%) developed antibodies against BVDV. In conclusion, the results indicate that BVDV is widespread in dairy herds in Majes, the high level of antibodies suggests the presence of PI animals in the bovine population and finally, bulk tank milk samples are very useful for antibody detection from milking cows.

Anticuerpos contra el virus de la diarrea viral bovina (VDVB) y antígeno del VDVB fueron evaluados en muestras de suero obtenidos en dos oportunidades, con intervalo de 30 días, en animales de 10 a 23 meses de edad (n = 148) de un establo de bovinos productores de leche que se encuentra en proceso de erradicación de la enfermedad de la diarrea viral bovina (DVB) desde 2002 . El establo está localizado en Santa Rita, Arequipa. Los anticuerpos y el antígeno viral se analizaron mediante las pruebas de neutralización viral y ELISA de captura, respectivamente. El 6.8% (10/148) de los animales presentaron anticuerpos contra el VDVB, existiendo un mayor número de animales con anticuerpos en el grupo etario de 19 a 23 meses de edad. Los títulos de anticuerpos variaron entre 8 a >256. No se detectaron animales portadores (PI) ni se encontró seroconversión en el segundo muestreo, lo que sugiere que los anticuerpos detectados correspondieron a infecciones pasadas; además, los resultados indican que la enfermedad de la DVB ha sido controlada en el hato., Antibodies against bovine viral diarrhea virus (BVDV) and antigen of BVDV wereevaluated in serum samples from cattle aged 10 to 23 months (n = 148) from a dairy herdwhere the disease eradication has been carrying out since 2002. The herd is located inSanta Rita, Arequipa, Perú, and samples were collected twice on a 30-day interval.Antibodies against BVDV and BVDV antigen were analyzed by viral neutralization andcapture ELISA tests respectively. Neutralising antibodies against BVDV were detected in6.8% (10/148) of animals, and most reactors were 19 to 23 months of age. The antibodytiter ranged from 8 to >256. None persistently infected (PI) animals were found. Theincidence of infection and seroconversion in the second set of samples was nill. Theseresults suggested that the antibodies against the BVD corresponded to residual antibodiesfrom previous infections. In addition, the results indicate the BVD infection has beencontrolled.

El objetivo del estudio fue evaluar la persistencia de los anticuerpos pasivos contra el virus de la Peste Porcina Clásica (vPPC) en lechones de dos granjas tecnificadas (A y B) con distintas estrategias de vacunación contra el vPPC. En la granja A, las marranas fueron vacunadas a los 90 días de gestación y en B a los 18 a 21 días post-parto. Se colectó muestras de sangre de lechones de ambas granjas durante la primera (n=15), tercera (n=15), quinta (n=15) y séptima (n=15) semana de edad, así como de 15 marranas por granja para la detección de los anticuerpos mediante la prueba de ELISA indirecta. Todos los lechones presentaron anticuerpos pasivos en la primera semana de edad, persistiendo en la mayoría de los lechones hasta la séptima semana de edad. Hubo diferencia significativa (p

Long-acting nanoformulated antiretroviral therapy (nanoART) induces a range of innate immune migratory, phagocytic and secretory cell functions that perpetuate drug depots. While recycling endosomes serve as the macrophage subcellular depots, little is known of the dynamics of nanoART-cell interactions. To this end, we assessed temporal leukocyte responses, drug uptake and distribution following both intraperitoneal and intramuscular injection of nanoformulated atazanavir (nanoATV). Local inflammatory responses heralded drug distribution to peritoneal cell populations, regional lymph nodes, spleen and liver. This proceeded for three days in male Balb/c mice. NanoATV-induced changes in myeloid populations were assessed by fluorescence-activated cell sorting (FACS) with CD45, CD3, CD11b, F4/80, and GR-1 antibodies. The localization of nanoATV within leukocyte cell subsets was determined by confocal microscopy. Combined FACS and ultra-performance liquid chromatography tandem mass-spectrometry assays determined nanoATV carriages by cell-based vehicles. A robust granulocyte, but not peritoneal macrophage nanoATV response paralleled zymosan A treatment. ATV levels were highest at sites of injection in peritoneal or muscle macrophages, dependent on the injection site. The spleen and liver served as nanoATV tissue depots while drug levels in lymph nodes were higher than those recorded in plasma. Dual polymer and cell labeling demonstrated a nearly exclusive drug reservoir in macrophages within the liver and spleen. Overall, nanoART induces innate immune responses coincident with rapid tissue macrophage distribution. Taken together, these works provide avenues for therapeutic development designed towards chemical eradication of human immunodeficiency viral infection.

Se determinó la prevalencia del virus de la diarrea viral bovina (VDVB) en bovinos mayores a seis meses de edad, entre hembras y machos (n = 406), pertenecientes a 114 pequeños criadores de tres comunidades de la cuenca de Ccañipía, provincia de Espinar, departamento del Cusco. La colección de sangre se realizó en tres grupos, según la edad Ɩ a 12, 13 a 23 y >24 meses de edad), para la detección de anticuerpos contra el VDVB, mediante la prueba de neutralización viral. El 56.2 ± 4.8% 鴤/406) de las muestras tuvieron anticuerpos contra el VDVB. No se detectaron animales portadores del virus. Animales con anticuerpos fueron encontrados en los tres grupos etarios pero el 65.4% 鳕/ 228) de serorreactores estuvieron en el grupo mayor de 24 meses. El 51.3% ྴ/39) de los toros jóvenes y adultos presentaron anticuerpos contra el VDVB. Los títulos de anticuerpos variaron entre 2 a mayor a 256. El 42.1% de animales entre 13 a >24 meses de edad tuvo títulos entre 128 a >256. El 86.8% ဃ/114) de los criadores tuvieron al menos un animal seropositivo al VDVB. The prevalence of bovine viral diarrhea virus (BVDV) was evaluated in 406 cattle of both sexes and older than 6 months. Animals belonged to 114 small farmers from three rural communities of the province of Espinar, Cusco, Peru. Blood samples were collected according to three age groups Ɩ-12, 13-23, >24 months old). Serum samples were tested for antibodies against BVDV using the viral neutralization test. The 56.2 ± 4.8% 鴤/406) of samples had antibodies against BVDV. Persistently infected animals were not detected. Antibodies were present in the three age groups, but the highest prevalence .4%) was detected in animals older than 24 months of age. The 51.3% ྴ/39) of young and adult bulls had antibodies against BVDV. Antibodies titers varied from 2 to >256, and high titers 鳀 to >256) were detected in 42.1% of animals of 13 to >24 months of age. The 86.8% ဃ/114) of the small farmers had at least one animal seropositive to BVDV.

El objetivo del presente estudio fue determinar el fenotipo y genotipo de cepas del Virus de la Diarrea Viral Bovina (VDVB) aisladas de bovinos de diversos lugares del Perú. Entre 1998 y 2007 se colectó muestras de tejidos de fetos abortados, sangre, suero o plasma de bovinos con infección aguda o portadores del virus procedente de las principales cuencas lecheras del país para la búsqueda del antígeno del VDVB mediante inmunofluorescencia y ELISA de captura. Las muestras positivas al antígeno viral fueron cultivadas en monocapas de células de cornete nasal de feto bovino (CNB), libre del VDVB hasta el tercer pasaje, para determinar el biotipo. Las muestras positivas, tanto de los tejidos como del tercer pasaje, fueron procesadas por la técnica RT-PCR en tiempo real, para determinar el genotipo viral. Se aislaron 41 cepas del VDVB en CNB. El 85.4% (35/41) correspondieron al fenotipo no citopático y 14.6% (6/41) al citopático. El 45.5% (25/55) y el 74.5% (41/55) de las muestras de tejidos y del 3er pasaje en CNB, respectivamente, resultaron positivas al VDVB mediante la técnica de RT-PCR en tiempo real. Todas las cepas pertenecieron al VDVB-1. Los resultados enriquecen la epidemiología de la DVB en el país y sugieren la ausencia o baja prevalencia de cepas del VDVB-2 en la población bovina de las principales cuencas lecheras., The aim of this study was to determine the phenotype and genotype of Bovine Viral Diarrhoea Virus (BVDV) in Peruvian cattle. During the period 1998-2007, tissues of aborted foetus, blood, serum or plasma samples of bovine with acute infection or persistently infected from dairy herds of various locations in the country were collected and the BVDV antigen was detected by immnunofluorescence or capture ELISA tests, and positive samples were inoculated en BVDV free BT cells and cultured until third passage to determine the biotype. Both animal tissues and third passage of positive samples were tested by a Real Time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) for genotyping the strains. Forty one BVDV strains were isolated in BT cells. Eighty five percent (35/41) of the isolates were non-cytopathic and 14.6% (6/41) cytophatic. Forty five and a half percent (25/55) and 74.5% (41/55) of the animal tissues and from the 3rd passage samples respectively, were positive to BVDV by Real Time RT-PCR test. All isolates belonged to BVDV-1 genotype. These results contribute to a better understanding of the epidemiology of the bovine viral diarrhoea disease in Peru and suggest that BVDV-2 is absent or has low prevalence in dairy herds.

The prevalence of bovine viral diarrhea virus (BVDV) infection was evaluated in 347 criollo cattle of both sexes and older than 6 months of age in small herds of the province of Melgar, Puno. Serum samples were tested for antibodies against BVDV using the viral neutralization test. The 48.7 ± 0.1% (166/347) of the samples had antibodies against BVDV. Animals carrying on the virus virus were not detected. Antibodies were detected in all sampled herds and the prevalence varied from 15.7 to 94.1%. Antibodies titers varied from 2 to >256, showing that BVDV infection is widespread in the local cattle population, suggesting the presence of factors promoting the viral distribution such livestock fairs and lack of control for animal movements within the region. El estudio tuvo por objetivo determinar la seroprevalencia del virus de la diarrea viral (VDVB) en bovinos criollos de la provincia de Melgar, Puno. Se obtuvieron muestras de sangre de bovinos (n= 347) machos y hembras mayores a seis meses de edad para la detección de anticuerpos neutralizantes mediante la prueba de neutralización viral. El 48.7 ± 0.1% (166/347) de los animales presentó anticuerpos contra el VDVB. No se detectaron animales portadores del virus. Anticuerpos fueron detectados en animales de todos los distritos con prevalencias entre 15.7 a 94.1%. Los títulos de anticuerpos estuvieron en un rango de 2 a >256, indicando que el VDVB está ampliamente difundido en bovinos de la zona. Las altas prevalencias de anticuerpos evidencian intensa actividad viral; así mismo, los títulos altos indican infecciones recientes y sugieren la existencia de factores que promueven la difusión viral como las ferias ganaderas y la falta de control en el tránsito interno de animales en la zona.

El objetivo del presente estudio fue determinar el efecto de la eliminación de los animales portadores del virus de la diarrea viral bovina (VDVB) sobre la seroconversión contra el virus en la nueva generación de animales de un establo lechero de crianza intensiva en Arequipa. Se colectaron muestras de suero a vaquillas entre 6 a 12 meses de edad en cuatro periodos: enero (n=73), junio (n=48) y octubre (n=48) del 2003 y enero del 2004 (n=35) para la detección de anticuerpos contra el VDVB y para la detección de animales portadores (PI) del virus, mediante las pruebas de neutralizaciónviral y ELISA captura de antigeno, respectivamente. La seroprevalencia del VDVB fue de 80.8 + 9.0, 56.3 + 14.0,50.0+ 14.2 y 22.9+ 13.9%en el primero, segundo, tercero y cuarto periodo de muestreo, respectivamente. La prevalencia de animales portadores del virus fue de 2.7% (2173) y fueron detectados únicamente en el grupo muestreado en enero del 2003. La incidencia de infección de VDVB fue 121100 (24159) al mes en el periodo enero 2003 a enero 2004. Mediante la pruebas de regresión logística se demostró que la eliminación de animales PI en enero del 2003 redujo el riesgo de infección en los animales susceptibles en los siguientes periodos de muestreo. Además, la edad mostró ser un factor de riesgo para la infección con VDVB. Los resultados indican que la infección con VDVB es altamente prevalente en hatos que albergan animales portadores y que su eliminación reduce el riesgo de infección en el resto de animales, como se describe en la literatura. Los resultados sugieren que es posible el control y erradicación de la DVB en hatos lecheros de crianza intensiva mediante la identificación y eliminación de los animales portadores y sin vacunación pero manteniendo un alto nivel de bioseguridad en el establo., A study was conducted to determine the effect of culling BVDV carrier animals on the seroconversion against BVDV in the new generation of heifers from a dairy herd located in Arequipa, Peru. Blood samples were collected to 6-12 month old females in four sampling periods: January (n=73), June (n=48), October 2003 (n=48), and January 2004 (n=35) to evaluate their serological status against BVDV and to screen for carrier animals using the virus neutralization and antigen-capture ELISA tests, respectively. The prevalence of BVDV was 80.8 ± 9.0, 56.3 ± 14.0, 50.0 ± 14.2 and 22.9 ± 13.9% in the first, second, third and fourth sampling period, respectively. There were 2.7% (2/73) of carrier heifers in the group sampled in January 2003, and none in the subsequent sampling periods. The incidence of BVDV infection was 12/100 heifers per month from January 2003 till January 2004. The logistic regression test showed that culling of carrier animals in January 2003 reduced the risk of infection in subsequent months. In addition, age was a risk factor for VDVB infection in this group of animals. The results showed that BVDV infection is highly prevalent in herds having carrier animals, and that the culling of PI animals reduce the risk of infection in herd mates as indicated in the literature. The results also suggest that the control and eradication of BVDV in intensive management dairy herds may be possible by identifying and culling carrier animals and without vaccination but ensuring high level of biosecurity.

The objetive of this study was to detect antibodies against vesicular stomatitis virus (VSV) in collared peccaries (Tayassu tajacu) from farms of Iquitos and Pucallpa, Peru. Blood samples were collected in animals olders than 6 months of age (pregnant females were not included) from a farm in Iquitos (n = 21) and from four farms in Pucallpa (n = 47), for detection of antibodies against Indiana 1 and New Jersey serotype of VSV by viral neutralization test. No specific antibodies against both serotypes of VSV were detected, indicating that the peccaries were not exposed to VSV, and suggesting absence of the infection in those animals., The objetive of this study was to detect antibodies against vesicular stomatitisvirus (VSV) in collared peccaries (Tayassu tajacu) from farms of Iquitos and Pucallpa,Peru. Blood samples were collected in animals olders than 6 months of age (pregnantfemales were not included) from a farm in Iquitos (n = 21) and from four farms in Pucallpa(n = 47), for detection of antibodies against Indiana 1 and New Jersey serotype of VSVby viral neutralization test. No specific antibodies against both serotypes of VSV weredetected, indicating that the peccaries were not exposed to VSV, and suggesting absenceof the infection in those animals.

Se describe un caso de aborto bovino debido a la bacteria Actinomyces pyogenes ocurrido en un hato lechero del valle de Cieneguilla, Lima, Perú. El feto de 4 meses de desarrollo fue abortado en completo estado de autolisis y fue remitido al laboratorio para su análisis. En la necropsia, la única lesión macroscópica observada fue de unos pequeños nódulos blanquecinos en el pulmón. Se colectó muestras de diferentes tejidos como riñón, pulmón, corazón y encéfalo para el descarte del virus de la diarrea viral bovina (VDVB) y Neospora caninum mediante pruebas inmunohistoquímicas y para el cultivo bacteriano. No se detectó antígenos del VDVB ni de N. caninum en los tejidos del feto, pero se aisló en cultivo puro una bacteria identificada bioquímicamente como A. pyogenes. El examen histopatológico del pulmón indicó una moderada a severa bronconeumonía supurativa aguda asociada a bacterias. El aislamiento de la bacteria en forma pura, la ausencia de otros agentes abortogénicos y las lesiones histopatológicas de los tejidos del feto indicaron que el aborto fue debido a A. pyogenes., A case of bovine fetal abortion due to Actinomyces pyogenes in a dairy herd of Cieneguilla valley of Lima, Peru is reported. A bovine fetus of four monts of gestation was aborted and submitted for laboratory examination. Macroscopic lesions in fetal tissues were not found with the exception of numerous white foci up to 1mm in diameter in diameter in the lungs. Various fetal tissue samples were collected for virologic, bacteriologic and histopathologic examinations. Bovine viral diarrhea virus (BVDV) and Neospora caninum antigen were ruled out by immunohistochemistry but, pure colonies of A. pyogenes were isolated from the lung and other fetal tissues. The microscopic examination showed moderate to severe bronchopneumonia associated to pyogenic bacterium. The isolation of A. pyogenes, the macro and microscopic lesions and the absence of BVDV an N. caninum antigen in the fetal tissues indicated the A. pyogenes as a causative agent of this fetal abortion.

Background Despite improved clinical outcomes seen following antiretroviral therapy (ART), resting CD4+ T cells continue to harbor latent human immunodeficiency virus type one (HIV-1). However, such cells are not likely the solitary viral reservoir and as such defining where and how others harbor virus is imperative for eradication measures. To such ends, we used HIV-1ADA-infected NOD.Cg-Prkdc scid Il2rg tm1Wjl/SzJ mice reconstituted with a human immune system to explore two long-acting ART regimens investigating their abilities to affect viral cell infection and latency. At 6 weeks of infection animals were divided into four groups. One received long-acting (LA) cabotegravir (CAB) and rilpivirine (RVP) (2ART), a second received LA CAB, lamivudine, abacavir and RVP (4ART), a third were left untreated and a fourth served as an uninfected control. After 4 weeks of LA ART treatment, blood, spleen and bone marrow (BM) cells were collected then phenotypically characterized. CD4+ T cell subsets, macrophages and hematopoietic progenitor cells were analyzed for HIV-1 nucleic acids by droplet digital PCR. Results Plasma viral loads were reduced by two log10 or to undetectable levels in the 2 and 4ART regimens, respectively. Numbers and distributions of CD4+ memory and regulatory T cells, macrophages and hematopoietic progenitor cells were significantly altered by HIV-1 infection and by both ART regimens. ART reduced viral DNA and RNA in all cell and tissue compartments. While memory cells were the dominant T cell reservoir, integrated HIV-1 DNA was also detected in the BM and spleen macrophages in both regimen-treated mice. Conclusion Despite vigorous ART regimens, HIV-1 DNA and RNA were easily detected in mature macrophages supporting their potential role as an infectious viral reservoir. Electronic supplementary material The online version of this article (doi:10.1186/s12977-017-0344-7) contains supplementary material, which is available to authorized users.