Surveillance of adverse events following immunization with meningococcal group C conjugate vaccine: Tuscany, 2005-2012

Introduction. Post-licensure vaccine safety studies are essential to identify uncommon events that may be difficult to assess during prelicensure studies. The aim of our study was to evaluate the safety of serogroup C meningococcal conjugate (MCC) vaccine in Tuscany from 2005 to 2012. Methods. All adverse events (AEs) to MCC vaccine notified from 2005 to 2012 were obtained from the regional health authority. Results. Following 451,570 doses administered, 110 suspected AEs were notified (mean annual reporting rate: 2.8/10,000 doses). The most frequently AE reported was fever (60%), fol- lowed by swelling at the injection site (11%) and febrile seizures (10%). Overall, 77.3% of cases were not severe, while 21.8% required hospitalization. Almost four months after the receipt of the vaccine, a one-year-old infant was diagnosed with a pervasive developmental disorder with disturbance of speech, but any link with the vaccinations received was refuted. Most AEs (80.9%) occurred after co-administration with other vaccines, especially with MMR or MMRV vaccines (42.7%) or the DTPa-HBV-IPV/ Hib vaccine (33.7%). Discussion. Our study confirmed the high level of safety of MCC vaccine in Tuscany: AEs proved rare and all cases had only tem- porary and self-resolving consequences. As usually only the most severe suspected AEs are reported, the true proportion of AEs requiring hospitalization was most probably overestimated, and it is plausible that most of these cases were in fact only temporally related

Recommended vaccinations for asplenic and hyposplenic adult patients

Asplenic or hyposplenic (AH) individuals are particularly vulnerable to invasive infections caused by encapsulated bacteria. Such infections have often a sudden onset and a fulminant course. Infectious diseases (IDs) incidence in AH subjects can be reduced by preventive measures such as vaccination. The aim of our work is to provide updated recommendations on prevention of infectious diseases in AH adult patients, and to supply a useful and practical tool to healthcare workers for the management of these subjects, in hospital setting and in outpatients consultation. A systematic literature review on evidence based measures for the prevention of IDs in adult AH patients was performed in 2015. Updated recommendations on available vaccines were consequently provided. Vaccinations against S. pneumoniae, N. meningitidis, H. influenzae type b and influenza virus are strongly recommended and should be administered at least 2 weeks before surgery in elective cases or at least 2 weeks after the surgical intervention in emergency cases. In subjects without evidence of immunity, 2 doses of live attenuated vaccines against measles-mumps-rubella and varicella should be administered 4–8 weeks apart from each other; a booster dose of tetanus, diphtheria and pertussis vaccine should be administered also to subjects fully vaccinated, and a 3-dose primary vaccination series is recommended in AH subjects with unknown or incomplete vaccination series (as in healthy people). Evidence based prevention data support the above recommendations to reduce the risk of infection in AH individuals

Proliferation and survival of human amniotic epithelial cells during their hepatic differentiation

Stem cells derived from placental tissues are an attractive source of cells for regenerative medicine. Amniotic epithelial cells isolated from human amnion (hAECs) have desirable and competitive characteristics that make them stand out between other stem cells. They have the ability to differentiate toward all three germ layers, they are not tumorigenic and they have immunosuppressive properties. Although liver transplantation is the best way to treat acute and chronic hepatic failure patients, there are several obstacles. Recently, stem cells have been spotlighted as alternative source of hepatocytes because of their potential for hepatogenic differentiation. In this work, we aimed to study the proliferation and survival of the hAECs during their hepatic differentiation. We have also analyzed the changes in pluripotency and hepatic markers. We differentiated amniotic cells applying a specific hepatic differentiation (HD) protocol. We determined by qRT-PCR that hAECs express significant levels of SOX-2, OCT-4 and NANOG during at least 15 days in culture and these pluripotent markers diminish during HD. SSEA-4 expression was reduced during HD, measured by immunofluorescence. Morphological characteristics became more similar to hepatic ones in differentiated cells and representative hepatic markers significantly augmented their expression, measured by qRT-PCR and Western blot. Cells achieved a differentiation efficiency of 75%. We observed that HD induced proliferation and promoted survival of hAECs, during 30 days in culture, evaluated by 3H-thymidine incorporation and MTT assay. HD also promoted changes in hAECs cell cycle. Cyclin D1 expression increased, while p21 and p53 levels were reduced. Immunofluorescence analysis showed that Ki-67 expression was upregulated during HD. Finally, ERK 1/2 phosphorylation, which is intimately linked to proliferation and cell survival, augmented during all HD process and the inhibition of this signaling pathway affected not only proliferation but also differentiation. Our results suggest that HD promotes proliferation and survival of hAECs, providing important evidence about the mechanisms governing their hepatic differentiation. We bring new knowledge concerning some of the optimal transplantation conditions for these hepatic like cells.Fil: Maymo, Julieta Lorena. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Riedel, Rodrigo Nicolas. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Pérez Alcázar, Germán Antonio. Hospital Universitario Virgen Macarena;Fil: Magatti, Marta. Istituto Ospedaliero;Fil: Maskin, Bernardo. Hospital Nacional Professor Dr. Alejandro Posadas; ArgentinaFil: Dueñas, José Luis. Hospital Universitario Virgen Macarena;Fil: Parolini, Ornella. Istituto Ospedaliero;Fil: Sánchez-Margalet, Víctor. Hospital Universitario Virgen Macarena;Fil: Varone, Cecilia Laura. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentin

Co-administration of vaccines: a focus on tetravalent Measles-Mumps-Rubella-Varicella (MMRV) and meningococcal C conjugate vaccines

Simultaneous administration of different vaccines is a strategy to increase the possibility to receive vaccines at appropriate age, safely and effectively, reducing the number of sessions and allowing a more acceptable integration of new vaccines into National Immunization Programs (NIPs). Co-administration can be performed when there are specific indications in the Summary of Product Characteristics (SmPC) of the vaccines; but, in absence of these indications, the practice is possible if there are no specific contraindications nor scientific evidence to discourage simultaneous administration. The aim of this work is to review the safety and efficacy of co-administration of the tetravalent measles, mumps, rubella, and varicella (MMRV) and the meningococcal C (Men C) conjugate vaccines after 12 months of age. Several studies demonstrated that MMRV and Men C conjugate vaccines can be administered concomitantly without a negative impact on the safety and immunogenicity of either vaccines, inducing highly immunogenic responses

Hospitalizations for pneumonia, invasive diseases and otitis in Tuscany (Italy), 2002-2014: Which was the impact of universal pneumococcal pediatric vaccination?

Streptococcus pneumoniae is the main causative organism of acute media otitis in children and meningitis and bacterial pneumonia in the community. Since 2008 in Tuscany, central Italy, the pneumococcal conjugate vaccine (7-valent vaccine, switched to 13-valent vaccine in 2010) was actively offered free of charge to all newborns. Aim of the study is to evaluate the impact of pneumococcal pediatric vaccination in the Tuscan population on hospitalizations potentially caused by S. pneumoniae, during pre-vaccination (PVP, 2002–2007) and vaccination period (VP, 2009–2014). We analyzed hospital discharge records (HDRs) of all hospitals in Tuscany from 2002 to 2014. Hospitalizations potentially due to pneumococcal diseases were 347, 221. The general hospitalization rate was 716/100,000 inhabitants during PVP and 753/100,000 in VP, with a decrease of 29.1% in the age-group 0–9 y (“target” of the vaccination program) and an increase of 75.7% in subjects >64 y of age. During VP, admission days and hospitalization costs increased (6.2% and 24.2%, respectively), especially in patients >64 y (12.9% and 33.8%, respectively); in children 64 y

Embryonic antigen SSEA-4 diminishes during HD of hAECs.

<p>Amniotic cells were seeded in 24-wells plate and incubated in IMDM 10% FBS (C) or hepatic differentiation medium (HD), during indicated times. At each time cells were fixed and SSEA-4 expression (green) was detected using FITC conjugated secondary antibody. Representative micrographs from hAECs at different HD times at 10X are shown. The nuclei were stained with DAPI (blue). Percentage of positive cells is shown on the left of images. Graph on the side shows cells fluorescence intensity for SSEA-4. Scale bar: 30 μm. Representative results from three replicates are shown. ***p<0.001 vs. respective control.</p

Hepatic differentiation increases proliferation and viability of amniotic cells.

<p><b>(A, B, C)</b> hAECs were cultivated in 6-wells plate with IMDM 10% FBS (C) or with hepatic differentiation medium (HD). 24 h before indicated times, ³H-thymidine (1 μCi/ml) was added and cells were cultured. Cell lysates were prepared and ³H-thymidine incorporation was determined as described in Materials and Methods. <b>(D, E)</b> Amniotic epithelial cells were seeded (5x10⁴) in 24-wells plate in complete IMDM medium supplemented with 10% FBS (C) or in hepatic differentiation medium (HD). Cells viability was determined by MTT test for up to 72 h <b>(D)</b> or during 30 days <b>(E). (F)</b> Amniotic cells were seeded in 24-wells plate and incubated in IMDM 10% FBS (C) or hepatic differentiation medium (HD), during indicated times. At each time cells were fixed and Ki-67 expression (green) was detected using Alexa-488 conjugated secondary antibody. Representative micrographs from hAECs at different HD times taken at 10X are shown. The nuclei were stained with DAPI (blue). <b>(G)</b> Graph shows the percentage of hAECs positive for Ki-67. Scale Bar: 100 μm. Results from a representative experiment are shown and are expressed as means ± S.D. for five independent experiments. *p<0.05; **p<0.01; ***p<0.001 vs. control.</p

ERK 1/2 phosphorylation is stimulated during hAECs hepatic differentiation.

<p><b>(A)</b> Amniotic cells were incubated during 1, 3, 10 (top), 15 and up to 30 days (bottom) with IMDM medium 10% FBS (C) or hepatic differentiation medium (HD), as indicated. Extracts from cells were prepared as previously described and loaded in a 12% SDS-PAGE. ERK 1/2 phosphorylation was determined by Western-blot. Loading controls were performed by immunoblotting the same membranes with anti-ERK 1/2 and anti-GAPDH. Bands densitometry is shown in lower panels. Molecular weight (kDa) is indicated at the right of the blot. <b>(B)</b> Amniotic cells were seeded in 24-wells plate and incubated in IMDM medium supplemented with 10% FBS (C) or hepatic differentiation medium (HD), during indicated times. At each time cells were fixed and ERK 1/2 phosphorylation (green) was detected using Alexa-488 conjugated secondary antibody. Representative micrographs at 10X from hAECs at different HD times are shown. The nuclei were stained with DAPI (blue). <b>(C)</b> Graph shows P-ERK fluorescence intensity in hAECs. Scale Bar: 30 μm. Results are expressed as mean ± S.D. for five independent experiments. *p< 0.05, **p< 0.01, ***p< 0.001 vs. control.</p