Research Center Ospedale Pediatrico Bambino Gesu

Rome, Italy

Research Center Ospedale Pediatrico Bambino Gesu

Rome, Italy

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Rosado M.M.,Research Center Ospedale Pediatrico Bambino Gesu | Scarsella M.,Research Center Ospedale Pediatrico Bambino Gesu | Pandolfi E.,Epidemiology Unit | Cascioli S.,Research Center Ospedale Pediatrico Bambino Gesu | And 10 more authors.
European Journal of Immunology | Year: 2011

The immunogenicity of a vaccine is conventionally measured through the level of serum Abs early after immunization, but to ensure protection specific Abs should be maintained long after primary vaccination. For hepatitis B, protective levels often decline over time, but breakthrough infections do not seem to occur. The aim of this study was to demonstrate whether, after hepatitis B vaccination, B-cell memory persists even when serum Abs decline. We compared the frequency of anti-hepatitis-specific memory B cells that remain in the blood of 99 children five years after priming with Infanrix®-hexa (GlaxoSmithKline) (n=34) or with Hexavac® (Sanofi Pasteur MSD) (n=65). These two vaccines differ in their ability to generate protective levels of IgG. Children with serum Abs under the protective level, <10mIU/mL, received a booster dose of hepatitis B vaccine, and memory B cells and serum Abs were measured 2wk later. We found that specific memory B cells had a similar frequency in all children independently of primary vaccine. Booster injection resulted in the increase of memory B cell frequencies (from 11.3 in 106 cells to 28.2 in 106 cells, p<0.01) and serum Abs (geometric mean concentration, GMC from 2.9 to 284mIU/mL), demonstrating that circulating memory B cells effectively respond to Ag challenge even when specific Abs fall under the protective threshold. © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Picchianti-Diamanti A.,University of Rome La Sapienza | Rosado M.M.,Research Center Ospedale Pediatrico Bambino Gesu | Scarsella M.,Research Center Ospedale Pediatrico Bambino Gesu | Lagana B.,University of Rome La Sapienza | D'Amelio R.,University of Rome La Sapienza
International Journal of Molecular Sciences | Year: 2014

Autoimmune diseases such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA) and psoriatic arthritis (PsA) are chronic inflammatory disorders of unknown etiology characterized by a wide range of abnormalities of the immune system that may compromise the function of several organs, such as kidney, heart, joints, brain and skin. Corticosteroids (CCS), synthetic and biologic immunosuppressive agents have demonstrated the capacity to improve the course of autoimmune diseases. However, a significant number of patients do not respond or develop resistance to these therapies over time. P-glycoprotein (P-gp) is a transmembrane protein that pumps several drugs out of the cell, including CCS and immunosuppressants; thus, its over-expression or hyper-function has been proposed as a possible mechanism of drug resistance in patients with autoimmune disorders. Recently, different authors have demonstrated that P-gp inhibitors, such as cyclosporine A (CsA) and its analogue Tacrolimus, are able to reduce P-gp expression and or function in SLE, RA and PsA patients. These observations suggest that P-gp antagonists could be adopted to revert drug resistance and improve disease outcome. The complex inter-relationship among drug resistance, P-gp expression and autoimmunity still remains elusive. © 2014 by the authors; licensee MDPI, Basel, Switzerland.


Scaldaferri M.L.,University of Rome Tor Vergata | Klinger F.G.,University of Rome Tor Vergata | Farini D.,University of Rome Tor Vergata | Di Carlo A.,University of Rome Tor Vergata | And 3 more authors.
Mechanisms of Development | Year: 2015

In the present paper, starting from the observation of heterogeneous expression of the GOF-18δPE-GFP Pou5f1 (Oct3/4) transgene in putative mouse PGC populations settled in the aorta-gonad-mesonephros (AGM) region, we identified various OCT3/4 positive populations showing distinct expression of PGC markers (BLIMP-1, AP, TG-1, STELLA) and co-expressing several proteins (CD-34, CD-41, FLK-1) and genes (Brachyury, Hox-B4, Scl/Tal-1 and Gata-2) of hematopoietic precursors. Moreover, we found that Oct3/4-GFPweak CD-34weak/high cells possess robust hematopoietic colony forming activity (CFU) in vitro. These data indicate that the cell population usually considered PGCs moving toward the gonadal ridges encompasses a subset of cells co-expressing several germ cell and hematopoietic markers and possessing hematopoietic activity. These results are discussed within of the current model of germline segregation. © 2015 Elsevier Ireland Ltd.


Diamanti A.P.,University of Rome La Sapienza | Rosado M.,Research Center Ospedale Pediatrico Bambino Gesu | Germano V.,University of Rome La Sapienza | Scarsella M.,Research Center Ospedale Pediatrico Bambino Gesu | And 5 more authors.
Clinical Immunology | Year: 2011

Secondary resistance may be a major problem in the management of autoimmune diseases. P-glycoprotein (P-gp) over-function has been described as a mechanism of drug resistance in autoimmune patients. P-gp function can in vitro be inhibited by cyclosporine A (CSA) and verapamil; moreover, P-gp reduction by CSA in systemic lupus erythematosus and rheumatoid arthritis has been demonstrated. Here, P-gp function before and after CSA administration in three psoriatic arthritis (PsA) patients, who developed a resistance to MTX/SSA, has been evaluated. P-gp function on patient cells was analyzed by measuring the changes in rhodamine-123 (Rh-123) fluorescence after verapamil incubation. CSA treatment resulted in good clinical outcome that was related with a significant P-gp function reduction at CD3+ and CD8+ levels. In addition to its immunosuppressive activity, CSA results may also be related to MTX/SSA effect restoration through P-gp inhibition. This is the first time that CSA has been demonstrated as being able to revert MTX/SSA resistance in PsA. © 2010 Elsevier Inc.


Rosado M.M.,Research Center Ospedale Pediatrico Bambino Gesu | Scarsella M.,Research Center Ospedale Pediatrico Bambino Gesu | Cascioli S.,Research Center Ospedale Pediatrico Bambino Gesu | Giorda E.,Research Center Ospedale Pediatrico Bambino Gesu | Carsetti R.,Research Center Ospedale Pediatrico Bambino Gesu
Methods in Molecular Biology | Year: 2013

Lymphocyte characterization is primarily based on the differential expression of surface markers. In this context, flow-cytometry analysis (FACS) is an exceptional technique that not only allows the identification of B-cell subsets, but can also be used to evaluate cell function, activation, and division. Here, we will combine the use of FACS analysis and ELISA techniques to identify murine bone marrow and peripheral B-cell subsets. The main function of B cells, derived through a multistage differentiation process from precursor cells, is to produce antibodies. This task is performed by terminally differentiated B cells called antibody-secreting cells (ASC) present at mucosal sites, in the bone marrow and in the spleen. The number and specificity of ASC can be measured by Enzyme-linked immunosorbent spot (ELISPOT) assay, a variation of the enzyme-linked immunosorbent assay (ELISA) used to quantify serum immunoglobulins. © 2013 Springer Science+Business Media, LLC.


PubMed | University of Rome Tor Vergata and Research Center Ospedale Pediatrico Bambino Gesu
Type: | Journal: Mechanisms of development | Year: 2015

In the present paper, starting from the observation of heterogeneous expression of the GOF-18PE-GFP Pou5f1 (Oct3/4) transgene in putative mouse PGC populations settled in the aorta-gonad-mesonephros (AGM) region, we identified various OCT3/4 positive populations showing distinct expression of PGC markers (BLIMP-1, AP, TG-1, STELLA) and co-expressing several proteins (CD-34, CD-41, FLK-1) and genes (Brachyury, Hox-B4, Scl/Tal-1 and Gata-2) of hematopoietic precursors. Moreover, we found that Oct3/4-GFP(weak) CD-34(weak/high) cells possess robust hematopoietic colony forming activity (CFU) in vitro. These data indicate that the cell population usually considered PGCs moving toward the gonadal ridges encompasses a subset of cells co-expressing several germ cell and hematopoietic markers and possessing hematopoietic activity. These results are discussed within of the current model of germline segregation.

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