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1.
J Phys Chem A ; 124(1): 108-125, 2020 Jan 09.
Article in English | MEDLINE | ID: mdl-31769686

ABSTRACT

Calculations of the photoionization cross section and asymmetry parameter, ß, are performed at the density functional theory (DFT) and time-dependent density functional theory (TDDFT) levels for all 32 valence levels of C60. Accurate numerical results are obtained for the isolated molecule in icosahedral symmetry. A detailed analysis based on the comparison between the DFT and TDDFT results allows the identification of four types of resonances: the well-known confinement resonances of mainly geometrical origin, shape resonances native to the ionization channel, induced shape resonances, and autoionization resonances brought about by interchannel coupling, as well as their different prominence in cross section or asymmetry parameter. Generally, cross sections are enhanced at the TDDFT level, which includes contribution from the bound-state excitations from closed channels, neglected at the DFT level, and the effect persists even well above the highest ionization threshold. This effect is best seen in the total cross section, although not as dramatic as found from simpler models, probably due to the stiffer electronic structure inherent in the full molecular description. The effects of interchannel coupling on individual native resonances are rather less predictable, leading to both enhancement and decreases and often altering the details of the structure significantly. A comparison with the previous accurate total cross-sectional calculations, as well as with the available experimental data, is very good for cross sections but slightly inferior for ß's. The results reported can serve as a reference to compare the effects of different environments on C60, as well as chemical substitution, notably endohedral fullerenes.

2.
Phys Chem Chem Phys ; 21(4): 1937-1951, 2019 Jan 23.
Article in English | MEDLINE | ID: mdl-30632573

ABSTRACT

This paper investigates the first sigma satellite band, which is by far the most prominent, in the valence photoelectron spectra for a set of isoelectronic diatomic molecules: carbon monoxide, carbon monosulfide, carbon monoselenide, silicon monoxide and boron monofluoride. In particular, we analyze the effect of the electronic structure, with the change of the atomic pair along the row and column of the periodic table on the position of the satellite peak as well as on the related dynamical observables profiles. For this investigation, highly correlated calculations have been performed on the primary ionic states and the satellite band for all the molecules considered. Cross sections for the primary ionic states, calculated using Dyson orbitals, have been compared with those obtained with Hartree-Fock and Density Functional Theory to probe the impact of the correlation in the bound states on the photoionization observables. Limitations of a simple intensity borrowing mechanism clearly result from the analysis of the satellite state, characterized by different features with respect to the relevant primary states.

3.
Nat Commun ; 9(1): 63, 2018 01 04.
Article in English | MEDLINE | ID: mdl-29302026

ABSTRACT

The first steps in photochemical processes, such as photosynthesis or animal vision, involve changes in electronic and geometric structure on extremely short time scales. Time-resolved photoelectron spectroscopy is a natural way to measure such changes, but has been hindered hitherto by limitations of available pulsed light sources in the vacuum-ultraviolet and soft X-ray spectral region, which have insufficient resolution in time and energy simultaneously. The unique combination of intensity, energy resolution, and femtosecond pulse duration of the FERMI-seeded free-electron laser can now provide exceptionally detailed information on photoexcitation-deexcitation and fragmentation in pump-probe experiments on the 50-femtosecond time scale. For the prototypical system acetylacetone we report here electron spectra measured as a function of time delay with enough spectral and time resolution to follow several photoexcited species through well-characterized individual steps, interpreted using state-of-the-art static and dynamics calculations. These results open the way for investigations of photochemical processes in unprecedented detail.

4.
J Phys Chem A ; 118(33): 6692-8, 2014 Aug 21.
Article in English | MEDLINE | ID: mdl-24856538

ABSTRACT

The present work concerns the study of high-energy structures in the photoionization of Mg and Be metallocenes due to photoelectron diffraction. The influence of geometrical structure is studied by varying the metal­ring distance in MgCp2, as well as that in the permethylated compounds MgCp2* and BeCp2*. The cross section ratios relative to the two outermost valence ionizations have been studied and found to be very sensitive to the value of the metal­ring distance and to be able to resolve ambiguities in present experimental values. Further differences are attributed to minor changes in the electronic structure. The results confirm that long-range oscillations in molecular photoemission cross sections constitute a general phenomenon and are an easily measurable observable that can be used to obtain important information on the geometric and electronic structure of the target.


Subject(s)
Electrons , Organometallic Compounds/chemistry , Photochemical Processes , Molecular Structure , Quantum Theory
5.
Phys Chem Chem Phys ; 15(4): 1310-8, 2013 Jan 28.
Article in English | MEDLINE | ID: mdl-23232557

ABSTRACT

The first experimental study of the X-ray absorption spectrum (XAS) of the allyl free radical, CH(2)CHCH(2), is reported. A supersonic He seeded beam of hyperthermal allyl radicals was crossed by a high resolution synchrotron radiation (SR) in the focus of a 3D ion momentum imaging time-of-flight (TOF) spectrometer to investigate the soft X-ray absorption and fragmentation processes. The XAS, recorded as Total-Ion-Yield (TIY), is dominated by C1s electron excitations from either the central carbon atom, C(C), or the two terminal carbon atoms, C(T), to the frontier orbitals, the semi-occupied-molecular-orbital (SOMO) and the lowest-unoccupied-molecular-orbital (LUMO). All of the intense features in the XAS could only be assigned with the aid of ab initio spectral simulation at the Multi-Configuration Self-Consistent-Field (MCSCF) level of theory, this level being required because of the multi-reference nature of the core-excited state wavefunctions of the open shell molecule. The ionization energies (IEs) of the singlet and triplet states of the C1s ionized allyl radical (XPS) were also calculated at the MCSCF level.

6.
Phys Chem Chem Phys ; 11(20): 3868-74, 2009 May 28.
Article in English | MEDLINE | ID: mdl-19440614

ABSTRACT

The formation of clusters of molecular hydrogen around a cationic charge, the Li(+) ion, is modelled by treating the global interaction as a sum of potentials where the Li(+)-H(2) forces come from a full anisotropic potential energy surface produced earlier in our group. The H(2)-H(2) interaction is taken from the literature and treated as a spherical potential between structureless bosonic solvent molecules of para-H(2) (pH(2)). The optimization of geometries and the minimum energy values are obtained via a genetic algorithm treatment whose structures are modified at the end to include a modelling of quantum effects. The results of hydrogen clustering around the cationic dopant indicate the presence of marked shell structures which are initially completed by the octahedral arrangement of the first six solvent partners, while the next shells are dominated by the mainly dispersive interaction among pH(2) molecules and show, in larger clusters, less structured solvent collocations around the ionic impurity.

7.
Panminerva Med ; 48(2): 119-27, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16953149

ABSTRACT

AIM: The human cytomegalovirus (HCMV) is an important pathogen in immunocompromised patients, such as transplant recipients. The use of sensitive and rapid diagnostic assays can have a great impact on antiviral prophylaxis and therapy monitoring and diagnosing active disease. Quantification of HCMV DNA may additionally have prognostic value and guide routine management. The aim of this study was to develop a reliable internally-controlled quantitative-competitive PCR (QC-PCR) for the detection and quantification of HCMV DNA viral load in peripheral blood and compare it with other methods: the HCMV pp65 antigenaemia assay in leukocyte fraction, the HCMV viraemia, both routinely employed in our laboratory, and the nucleic acid sequence-based amplification (NASBA) for detection of HCMV pp67-mRNA. METHODS: Quantitative-competitive PCR is a procedure for nucleic acid quantification based on co-amplification of competitive templates, the target DNA and a competitor functioning as internal standard. In particular, a standard curve is generated by amplifying 10(2) to 10(5) copies of target pCMV-435 plasmid with 10(4) copies of competitor pCMV-C plasmid. Clinical samples derived from 40 kidney transplant patients were tested by spiking 10(4) copies of pCMV-C into the PCR mix as internal control, and comparing results with the standard curve. RESULTS: Of the 40 patients studied, 39 (97.5%) were positive for HCMV DNA by QC-PCR. While the correlation between the number of pp65-positive cells and the number of HCMV DNA genome copies/mL and the former and the pp67mRNA-positivity were statistically significant, there was no significant correlation between HCMV DNA viral load assayed by QC-PCR and HCMV viraemia. CONCLUSIONS: The QC-PCR assay could detect from 10(2) to over 10(7) copies of HCMV DNA with a range of linearity between 10(2) and 10(5) genomes.


Subject(s)
Antigens, Viral/blood , Cytomegalovirus/isolation & purification , Phosphoproteins/blood , Phosphoproteins/genetics , Polymerase Chain Reaction/methods , RNA, Messenger/analysis , Viral Matrix Proteins/blood , Viral Matrix Proteins/genetics , Viremia/diagnosis , Adult , Aged , DNA, Viral/blood , Female , Humans , Male , Middle Aged , Nucleic Acid Amplification Techniques , Viral Load
8.
Panminerva Med ; 48(1): 59-66, 2006 Mar.
Article in English | MEDLINE | ID: mdl-16633333

ABSTRACT

AIM: Quantitative polymerase chain reaction (PCR) analysis to evaluate virus load in comparison with the patient's base-line virus levels would be an optimal diagnostic approach to monitoring human polyomavirus infections and to investigate their possible involvement in the onset of nephropathy in this patient group. Studies on the correlation between viral burden and renal disease have pointed to the incidence of JC virus (JCV) related progressive multifocal leukoencephalopathy (PML) occurring in renal and haematopoietic stem cell transplant recipients. METHODS: We developed a reliable internally-controlled quantitative PCR assay to measure JCV-DNA in fluid samples of urine, serum and cerebrospinal fluid (CSF) by densitometric analysis of the amplification products. The assay was also used to evaluate the JCV load in CFS samples from patients with suspected demyelinating syndrome and in urine and serum samples from healthy subjects and renal transplant recipients. RESULTS: All CSF samples from the 51 patients with suspected demyelinating syndrome tested JCV-DNA negative: none of them had a diagnosed PML. Analysis of the prevalence of JCV-viruria and JCV-viraemia confirmed our previous data. JCV-viruria was detected in 17% of renal transplant recipients and 26.6% of healthy controls; JCV-viraemia was found in 3.4% of transplant patients and 0% in controls. Noteworthy was a lower prevalence of JCV-viraemia in the 116 (3.4%) renal transplant patients than the prevalence previously reported for the 51 (11.8%) patients with suspected demyelinating syndrome. The mean viral load of viruria was much higher in the healthy controls than in the transplant recipients [104020 DNA copies/mL (DS+/-62284) vs 4136 DNA copies/mL (DS+/-77371)]. CONCLUSIONS: The quantitative PCR assay developed in our lab offers in 2 h time a reliable true quantification of viral DNA by densitometric analysis of the amplification product. To check for the possible presence of potential Taq polymerase inhibitors an internal control (the homemade pJCV-C plasmid) is used. The relation between polyomavirus infections and their possible involvement in post-transplant pathologies need further investigation. It would be useful to monitor the JCV-DNA load in urine and serum from more renal transplant recipients, including patients with nephropathy or active graft rejection over a longer period of time.


Subject(s)
DNA, Viral/genetics , JC Virus/genetics , Polymerase Chain Reaction/methods , Base Sequence , Case-Control Studies , DNA, Viral/analysis , DNA, Viral/cerebrospinal fluid , Humans , JC Virus/isolation & purification , Kidney Transplantation , Leukoencephalopathy, Progressive Multifocal/virology , Polymerase Chain Reaction/standards
9.
Panminerva Med ; 46(3): 153-9, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15510083

ABSTRACT

AIM: Several studies have disclosed a correlation between human polyomavirus BK (BKV) and interstitial nephritis in renal transplant recipients. It has recently been hypothesized that some cases of nephropathy may be associated with human polyomavirus JC (JCV). METHODS: In this paper we describe the development of duplex nested-PCR assay which allows the simultaneous detection and discrimination of genomic sequences of JCV and BKV ''large T antigen'', resulting in amplicons of 150 bp and 278 bp, respectively. Thus, the presence of JCV and BKV DNA in urine and serum samples from 51 renal transplant recipients and 29 healthy controls was investigated and related to immunosuppressive regimens and renal function. RESULTS: The comparison between the incidence of the of BKV and/or JCV infections (detected by viruria and/or viraemia) in renal transplant recipients and the control group revealed a highly significant increase of the incidence of BKV infection in immunosuppressed patients vs healthy subjects (62.7% vs 27.6%; p=0.005). In particular, we found a significant increase of BKV-DNA viruria in renal transplant recipients vs healthy subjects (49% vs 17.2%; p=0.01), in agreement with the BKV urinary shedding in renal transplant recipients of the literature (5-45%). CONCLUSION: The nested-PCR technique is a valid diagnostic tool to detect viral presence in urine and its systemic diffusion. Our assay links the high sensitivity of nested amplification with the simultaneous detection and discrimination of genomic sequences of JC and BK polyomaviruses and thus provides a handy, rapid and sensitive means for DNA analysis of large numbers of samples.


Subject(s)
BK Virus/genetics , DNA, Viral/blood , DNA, Viral/urine , JC Virus/genetics , Kidney Transplantation , Adult , Case-Control Studies , Female , Humans , Incidence , Male , Middle Aged , Polymerase Chain Reaction , Polyomavirus Infections/epidemiology , Sensitivity and Specificity , Tumor Virus Infections/epidemiology
10.
New Microbiol ; 26(2): 141-9, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12737195

ABSTRACT

Post-transplant lymphoproliferative disorders (PTLD), ranging from lymphoid hyperplasia to clonal malignancy, are a severe complication arising in solid organ transplant patients. Their reported incidence ranges from 1 to 20%, according to factors such as type of transplanted organ and the age of recipients. A strong correlation between Epstein Barr virus (EBV) infection, the grade and type of immunosuppression and the development of PTLD has been recognized. The detection and quantification of EBV-DNA load in peripheral blood have been utilized as prognostic markers for the development of PTLD, showing a correlation between high levels of EBV-DNA in the blood and the development of PTLD. In this study, we monitored EBV viral load monthly in 15 renal transplant recipients for six months. The number of EBV-DNA copies was measured in peripheral blood mononuclear cells (PBMC) and serum samples by a quantitative PCR protocol developed in our laboratory that employes a previous screening of samples containing a significant number of viral DNA copies (> or =1000 copies/10(5) PBMC or 100 microl serum) by semi-quantitative PCR followed by a precise quantification of the only significant samples by quantitative-competitive (QC)-PCR. Our 15 renal transplant patients neither developed PTLD nor had recurrent acute illnesses or acute graft rejections during the study. The results obtained in the monthly follow up of EB viral load in PBMC samples confirmed its fluctuation in asymptomatic patients reported in literature. In particular, 5/14 (35.7%) of EBV seropositive patients had an EBV-DNA load equal to 1000 EBV copies /10(5) PBMC (roughly corresponding to 10.000 copies/microg PBMC DNA), and 1/14 (7.1%) reached 5000 EBV copies /10(5) PBMC (roughly corresponding to 50.000 copies/microg PBMC DNA), at least once in our study. In the EBV seronegative patient, EBV-DNA in PBMC samples was always undetectable (less than 100 DNA copies/10(5) PBMC). EBV-DNA load in all serum samples was less than threshold value of our quantification protocol (<100 DNA copies/100 microl serum), supporting the literature data. With regard to immunosuppressive treatment, 66.7% of the six patients in whom EBV load reached values equal to or higher than 1000 DNA copies/10(5) PBMC, were on FK506 whereas only 33.3% of them were on CyA. In conclusion, further investigations are needed to better understand the role of EBV infection in the pathogenesis of PTLD in immunosuppressed patients. Given the high positive predictive value of EB viral load in peripheral blood for diagnosis of PTLD reported by several authors, and the described absence of correlation between the serological evidence of EBV reactivation and EB viral load, EBV viral load measurement in PBMC and serum samples using quantitative PCR techniques is a powerful diagnostic tool to monitor transplanted patients at risk to develop PTLD.


Subject(s)
Epstein-Barr Virus Infections/diagnosis , Herpesvirus 4, Human/isolation & purification , Kidney Transplantation/adverse effects , Lymphoproliferative Disorders/diagnosis , Polymerase Chain Reaction/methods , Viral Load/methods , Cells, Cultured , DNA, Viral/analysis , DNA, Viral/blood , Epstein-Barr Virus Infections/etiology , Epstein-Barr Virus Infections/immunology , Female , Herpesvirus 4, Human/genetics , Humans , Lymphoproliferative Disorders/virology , Male , Time Factors
11.
J Clin Virol ; 26(3): 361-8, 2003 Apr.
Article in English | MEDLINE | ID: mdl-12637086

ABSTRACT

BACKGROUND: B19 virus infection with persistent anaemia has been reported in organ transplant recipients. Detection of B19 virus DNA in serum is the best direct marker of active infection. OBJECTIVE: The present study evaluated the incidence and clinical role of active B19 virus infection in renal transplant recipients presenting with anaemia. STUDY DESIGN: Forty-eight such recipients were investigated by nested PCR on serum samples. The controls were 21 recipients without anaemia. Active HCMV infection was also investigated as a marker of high immunosuppression. RESULTS AND CONCLUSIONS: In 11/48 (23%) patients B19 virus DNA was demonstrated in serum versus only 1/21 (5%) of the controls. Ten of these 11 patients had already been seropositive at transplantation and active infection occurred in eight of them during the first 3 months after transplantation. The remaining patient experienced a primary infection 9 months after transplantation. Eight (73%) of these 11 patients displayed a concomitant HCMV infection and four (36%) showed increasing serum creatinine levels but none developed glomerulopathy; 3/11 (27%) recovered spontaneously from anaemia whereas 8/11 (73%) needed therapy. In conclusion, the relatively high occurrence (23%) of B19 virus infection in patients presenting with anaemia, suggests that it should be considered in the differential diagnosis of persistent anaemia in renal transplant recipients. Presence of the viral DNA should be assessed early from transplantation and the viral load should be monitored to follow persistent infection and better understand the relation between active infection and occurrence of anaemia, and to assess the efficacy of IVIG therapy and/or immunosuppression reduction in clearing the virus.


Subject(s)
Anemia/etiology , Cytomegalovirus Infections/virology , DNA, Viral/blood , Kidney Transplantation , Parvoviridae Infections/virology , Parvovirus B19, Human/isolation & purification , Postoperative Complications/virology , Recombinant Fusion Proteins , Viremia/virology , Anemia/virology , Antibodies, Monoclonal/adverse effects , Antibodies, Viral/blood , Antilymphocyte Serum/adverse effects , Basiliximab , Cyclosporine/adverse effects , Cytomegalovirus Infections/etiology , Cytomegalovirus Infections/therapy , DNA, Viral/isolation & purification , Diagnosis, Differential , Disease Susceptibility , Female , Humans , Immunoglobulins, Intravenous/therapeutic use , Immunosuppressive Agents/adverse effects , Interleukin-1/antagonists & inhibitors , Male , Mycophenolic Acid/adverse effects , Mycophenolic Acid/analogs & derivatives , Parvoviridae Infections/etiology , Parvoviridae Infections/therapy , Parvovirus B19, Human/genetics , Parvovirus B19, Human/immunology , Phosphoproteins/blood , Polymerase Chain Reaction , Prednisone/adverse effects , Retrospective Studies , T-Lymphocytes , Tacrolimus/adverse effects , Viral Load , Viral Matrix Proteins/blood , Zidovudine/adverse effects
12.
Infect Immun ; 69(10): 5974-80, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11553533

ABSTRACT

Bartonella henselae is the causative agent of cat scratch disease (CSD), a self-limiting condition characterized by a subacute regional lymphadenopathy that may develop into disseminated bartonellosis in immunocompromised subjects. Mice experimentally infected with B. henselae display typical liver and spleen granulomas rich in T cells and macrophages. So far there are no data on the interaction between bartonellae and macrophages. In order to clarify this topic, we investigated the interaction of B. henselae with J774, a mouse macrophage cell line. Analysis of bacterial uptake by functional assays and transmission electron microscopy indicates that bartonellae can enter and survive inside J774. Entry occurred within 30 min postinfection and reached a plateau at 160 min. Infection of J774 was followed by a dose-dependent release of the proinflammatory cytokines tumor necrosis factor alpha, interleukin 1beta (IL-1beta), and IL-6. Bartonellae persisted intracellularly without loss of viability for at least 8 h, and their number slightly decreased 24 h postinfection. Gamma interferon (IFN-gamma) treatment of J774 significantly decreased the number of recoverable bacteria at 8 and 24 h. This enhancement of macrophage bactericidal activity was associated with nitric oxide (NO) release and was prevented by the addition of the competitive inhibitor of NO synthesis N(G)-monomethyl L-arginine. These findings suggest that IFN-gamma-mediated activation of macrophages may be important for the clearing of B. henselae infection and that anti-B. henselae microbicidal activity of IFN-gamma-activated macrophages is mediated to a large extent by NO production.


Subject(s)
Bartonella henselae/immunology , Macrophages/immunology , Animals , Bartonella henselae/physiology , Cell Line , Interferon-gamma/immunology , Interleukin-1/biosynthesis , Interleukin-6/biosynthesis , Kinetics , Macrophage Activation , Macrophages/microbiology , Mice , Nitric Oxide/biosynthesis , Tumor Necrosis Factor-alpha/biosynthesis
13.
New Microbiol ; 24(3): 223-9, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11497078

ABSTRACT

In this study we investigated the levels of Epstein Barr virus (EBV) DNA by quantitative polymerase chain reaction (Q-PCR) in serum, whole blood and peripheral blood mononuclear cells (PBMC) from anti-EA IgG seropositive or anti-EA IgG seronegative EBV infected renal transplant recipients. We compared serological data with the viral load to monitor the risk of developing post-transplant lymphoproliferative disorders (PTLD). All patients were asymptomatic and none of them developed PTLD at the time of the study. EBV DNA quantitation for each patient varied in whole blood and PBMC samples probably due to different numbers of mononuclear cells present in samples from which DNA was extracted (whole blood vs. purified PBMC). In 92% of the serum samples EBV DNA was undetectable probably due to absence of free genomes since the number of DNA copies detected in samples from whole blood and PBMC does not reach very high levels. The correlation between the presence of EA-antibody, considered serological evidence of EBV reactivation, and the viral load showed that 60% of EA-positive patients had quantifiable EBV DNA, whereas in 40% of EA-positive patients EBV DNA was undetectable, showing serological reactivity but no viral replication. Of the remaining EA-negative patients, EBV DNA could be detected in 71% of them, whereas 29% did not show EBV DNA, indicating no EBV replication. In conclusion, our results confirm that the presence of serum IgG anti-EA antibody is not a reliable marker of active EBV infection whereas the evaluation of the viral load in blood samples is a useful diagnostic tool to monitor and to better understand the course of EBV infection in immunocompromised renal transplant patients at risk of developing PTLD.


Subject(s)
Epstein-Barr Virus Infections/complications , Epstein-Barr Virus Infections/virology , Herpesvirus 4, Human/genetics , Herpesvirus 4, Human/isolation & purification , Kidney Transplantation , Viral Load , Adult , Antibodies, Viral/blood , DNA, Viral/blood , Epstein-Barr Virus Infections/immunology , Female , Herpesvirus 4, Human/immunology , Humans , Immunoglobulin G/blood , Kidney Transplantation/adverse effects , Leukocytes, Mononuclear/virology , Male , Middle Aged , Polymerase Chain Reaction , Risk Factors , Tissue Donors
14.
J Leukoc Biol ; 67(3): 381-6, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10733099

ABSTRACT

Serum amyloid A (SAA) is a 12-kDa protein secreted in large amounts by liver cells during microbial infections or inflammatory diseases. We have recently reported that SAA induces chemotaxis of polymorphonuclear cells (PMN), monocytes, and T lymphocytes and stimulates their adhesion to endothelial monolayers. In this study, we investigated whether SAA regulates PMN antimicrobial activities. We found that recombinant SAA (rSAA), at concentrations comparable to serum levels attained during an acute phase response, is a potent activator of PMN. Stimulation of PMN by rSAA results in a rapid and transient increase of cytosolic calcium concentration and up-regulation of cell-surface expression of antigens involved in adhesion and microbial recognition such as CD11c and CD16. In addition, stimulation of PMN with rSAA increases secretion of lactoferrin, an antimicrobial protein that is contained in specific granules of PMN and enhances PMN phagocytic activity against heat-killed Candida albicans. Finally, activation of PMN with rSAA enhances their anti-Candida activity within 30 min of stimulation. These results suggest that SAA is involved in up-regulating PMN antimicrobial activities and that high circulating concentrations of SAA as seen in the acute phase response may constitute a potential host defense mechanism against fungal infections.


Subject(s)
Apolipoproteins/pharmacology , Candida albicans/immunology , Cell Degranulation/drug effects , Neutrophil Activation , Neutrophils/immunology , Phagocytosis/drug effects , Serum Amyloid A Protein/pharmacology , Acute-Phase Reaction , Adult , Antigens, CD/metabolism , Calcium/metabolism , Candida albicans/cytology , Candida albicans/drug effects , Cells, Cultured , Dose-Response Relationship, Drug , Humans , Lactoferrin/metabolism , N-Formylmethionine Leucyl-Phenylalanine/pharmacology , Neutrophils/drug effects , Neutrophils/metabolism , Neutrophils/microbiology , Recombinant Proteins/pharmacology , Respiratory Burst/drug effects , Signal Transduction/drug effects , Up-Regulation/drug effects , Virulence Factors, Bordetella/pharmacology
15.
J Invest Dermatol ; 113(2): 238-45, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10469310

ABSTRACT

Hypertrophic scarring is a skin disorder that occurs after wounding and thermal injury. There is accumulating evidence that immunologic processes such as infiltration of activated T lymphocytes and altered cytokine production may play a role in the formation of hypertrophic scars. Interleukin-15, a cytokine identified as a T cell growth factor, also acts as a chemoattractant for T cells and has pro-inflammatory properties. We investigated the expression and the role of this cytokine in hypertrophic scarring. IL-15 expression was compared in skin biopsies of hypertrophic scars (HS) both in active (AHS) and in remission (RHS) phases, in normotrophic scars (NTS) and in normal skin using reverse transcriptase-polymerase chain reaction and immunohistochemistry. IL-15 expression in HS was significantly higher than in NTS or normal skin. Furthermore, AHS expressed higher levels of IL-15 than RHS. Immunohistologic analysis of AHS samples showed strong IL-15 immunoreactivity in keratinocytes and Langerhans cells in the epidermis and in macrophages, fibroblasts, and dermal dendritic cells in the dermis. High levels of IL-15 expression in AHS correlated with abundant infiltration of activated CD3+ cells. Ex vivo experiments indicate that IL-15 can sustain the proliferative response of T cells derived from AHS but not from RHS and NTS. In addition, IL-15 prevents both cytokine deprivation and activation-induced apoptosis of T cells derived from AHS. Taken together, these results suggest that IL-15 can be involved in the recruitment, proliferation, and apoptosis inhibition of T cells in AHS. The findings that the evolution from an AHS to a RHS is associated with a decrease in IL15 expression, and with a loss of IL-15 responsiveness in ex vivo-cultured T cells, indicate that this cytokine plays an important role in the biology of pathologic scar formation.


Subject(s)
Cicatrix, Hypertrophic/genetics , Interleukin-15/genetics , Interleukin-15/physiology , Adolescent , Adult , Aged , Apoptosis/drug effects , CD3 Complex , Cell Cycle/drug effects , Cell Division/drug effects , Cicatrix, Hypertrophic/metabolism , Cicatrix, Hypertrophic/pathology , Female , Gene Expression , Humans , Immunohistochemistry , Interleukin-15/pharmacology , Interleukin-2/pharmacology , Lymphocyte Count , Male , Middle Aged , Proto-Oncogene Proteins c-bcl-2/biosynthesis , Proto-Oncogene Proteins c-bcl-2/drug effects , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , T-Lymphocytes/cytology , T-Lymphocytes/immunology
16.
Blood ; 93(10): 3531-9, 1999 May 15.
Article in English | MEDLINE | ID: mdl-10233906

ABSTRACT

Interleukin-15 (IL-15) is a potent regulator of T-, B-, and natural killer cell proliferation and displays unusually tight controls of secretion. Even though IL-15 mRNA is constitutively expressed in monocytes/macrophages and is upregulated by a variety of stimuli, evidence for IL-15 cytokine secretion is only found exceptionally, eg, conditions of pathological, chronic inflammation. This raises the possibility that monocytes express membrane-bound IL-15 rather than secrete it. The current study explores this hypothesis. We demonstrate here that biologically active IL-15 is indeed detectable in a constitutively expressed, membrane-bound form on normal human monocytes, as well as on monocytic cell lines (MONO-MAC-6, THP-1, and U937), but not on human T or B cells (MT4, M9, C5966, JURKAT, DAUDI, RAJI, and Epstein-Barr virus-immortalized B-cell clones). Furthermore, cell surface-bound IL-15 is upregulated upon interferon-gamma stimulation. Interestingly, monocyte/macrophage inhibitory cytokines such as IL-4 and IL-13 fail to downregulate both constitutive and induced cell-surface expression of IL-15. Membrane-bound IL-15 does not elute with acetate buffer or trypsin treatment, suggesting that it is an integral membrane protein and that it is not associated with the IL-15 receptor complex. Finally, membrane-bound IL-15 stimulates T lymphocytes to proliferate in vitro, indicating that it is biologically active. These findings enlist IL-15 in the fairly small family of cytokines for which the presence of a biologically active membrane-bound form has been demonstrated (eg, IL-1, tumor necrosis factor-alpha, and IL-10) and invites the speculation that most of the biological effects of IL-15 under physiological conditions are exerted by the cell surface-bound form.


Subject(s)
Gene Expression Regulation/immunology , Interferon-gamma/pharmacology , Interleukin-15/genetics , Monocytes/immunology , Cells, Cultured , Flow Cytometry , Gene Expression Regulation/drug effects , Humans , Interleukin-10/pharmacology , Interleukin-13/pharmacology , Interleukin-15/blood , Interleukin-4/pharmacology , Jurkat Cells , Lipopolysaccharides/pharmacology , Monocytes/cytology , Recombinant Proteins/pharmacology , Tumor Cells, Cultured , U937 Cells
17.
Infect Immun ; 66(6): 2640-7, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9596728

ABSTRACT

Interleukin-15 (IL-15) is a recently discovered cytokine produced by a wide range of different cell types including fibroblasts, keratinocytes, endothelial cells, and macrophages in response to lipopolysaccharide or microbial infection. This suggests that IL-15 may play a crucial role in the activation of phagocytic cells against pathogens. We studied polymorphonuclear leukocyte (PMN) activation by IL-15, evaluated as enhancement of PMN anti-Candida activity as well as IL-8 production, following stimulation with the cytokine. The PMN response to IL-15 depends on binding to the IL-15 receptor. Our experiments show that binding of a biotinylated human IL-15-immunoglobulin G2b IgG2b fusion protein was competed by the addition of human recombinant IL-15 (rIL-15) or of human rIL-2, suggesting that IL-15 binding to PMN might involve the IL-2Rbeta and IL-2Rgamma chains, which have been shown to be constitutively expressed by PMN. In addition, we show by reverse transcription-PCR and by flow cytometry with a specific anti-IL-15Ralpha chain monoclonal antibody that PMN express the IL-15Ralpha chain at the mRNA and protein levels. Incubation with IL-15 activated PMN to secrete the chemotactic factor IL-8, and the amount secreted was increased by costimulation with heat-inactivated Candida albicans. In addition, IL-15 primed the metabolic burst of PMN in response to formyl-methionyl-leucyl-phenylalanine but was not sufficient to trigger the respiratory burst or to increase the production of superoxide in PMN exposed to C. albicans. IL-15 also increased the ability of PMN to phagocytose heat-killed C. albicans organisms in a dose-dependent manner, without opsonization by antibodies or complement-derived products. In the same concentration range, IL-15 was as effective as gamma interferon (IFN-gamma) and IL-2 in increasing the C. albicans growth-inhibitory activity of PMN. Taken together, these results suggest that IL-15 is a potent stimulant of both proinflammatory and antifungal activities of PMN, activating several antimicrobial functions of PMN involved in the cellular response against C. albicans.


Subject(s)
Candida albicans/immunology , Interleukin-15/pharmacology , Neutrophils/drug effects , Humans , Interleukin-15/metabolism , Interleukin-8/metabolism , Neutrophil Activation , Neutrophils/immunology , Phagocytosis/drug effects , Protein Binding , Superoxides/metabolism
18.
Blood ; 90(7): 2804-9, 1997 Oct 01.
Article in English | MEDLINE | ID: mdl-9326248

ABSTRACT

Interleukin-15 (IL-15) is a recently characterized cytokine that shares many biological activities with IL-2 and interacts with the beta and gamma components of the IL-2 receptor. Unlike IL-2, which is secreted only by T cells, IL-15 is expressed preferentially by nonlymphoid tissues, epithelial, and fibroblast cell lines and by activated monocytes/macrophages. High concentrations of IL-15 have been shown in inflamed joints of rheumatoid arthritis patients, suggesting a role for IL-15 in inflammatory diseases where there is recruitment of leukocytes. Although monocytes have been shown to bind IL-15, its effects on these cells are not defined. In this report we show that supernatants of monocytes treated with IL-15-contained chemotactic activity for neutrophils and monocytes which was neutralized by anti-IL-8 or by anti-monocyte chemotactic protein 1 (MCP-1) antibodies, respectively. Secretion of IL-8 and MCP-1 proteins is detectable by enzyme-linked immunosorbent assay as early as 6 hours after stimulation with IL-15. Production of the two chemokines is correlated with induction by IL-15 of mRNA expression in monocytes. In addition, IL-8 and MCP-1 induction by IL-15 is differently regulated by interferon-gamma (IFN-gamma) and IL-4. IFN-gamma inhibited IL-15-induced IL-8 secretion, but synergized with IL-15 in MCP-1 induction; whereas IL-4 inhibited both IL-8 and MCP-1 induction by IL-15. These results show that IL-15 can stimulate monocytes to produce chemokines that cause inflammatory cell accumulation. Thus, IL-15 locally produced at sites of inflammation may play a pivotal role in the regulation of the leukocyte infiltrate.


Subject(s)
Chemokine CCL2/biosynthesis , Gene Expression Regulation/drug effects , Interleukin-15/pharmacology , Interleukin-8/biosynthesis , Monocytes/drug effects , Cells, Cultured , Chemokine CCL2/genetics , Chemokine CCL2/metabolism , Chemotaxis, Leukocyte/drug effects , Humans , Inflammation/metabolism , Interferon-gamma/pharmacology , Interleukin-4/pharmacology , Interleukin-8/genetics , Interleukin-8/metabolism , Monocytes/metabolism , RNA, Messenger/biosynthesis , Recombinant Proteins/pharmacology , Stimulation, Chemical , T-Lymphocytes/immunology , T-Lymphocytes/metabolism
19.
Minerva Urol Nefrol ; 48(3): 139-43, 1996 Sep.
Article in Italian | MEDLINE | ID: mdl-8966650

ABSTRACT

We have studied EBV infection in renal transplant patients during the first year after transplantation. At trasplantation all patients were EBV seropositive and reactivation of EBV infection was demonstrated in 54% cases after one year. CMV active infection was also demonstrated in 42% of patients with EBV reactivation. No correlation was observed between EBV reactivation and age, sex, immunosuppressive treatment, degree of immunosuppression or donor/recipient HLA matching. A correlation between immunosuppressive treatment, EBV infection and lymphoproliferative disorders (LD) is described in literature, however none of our patients developed LD so far, probably due to the different immunosuppressive protocol employed.


Subject(s)
Cytomegalovirus Infections/therapy , Herpesviridae Infections/therapy , Herpesvirus 4, Human/isolation & purification , Kidney Transplantation , Lymphoproliferative Disorders/therapy , Adult , Antibodies, Viral/immunology , Cyclosporine/therapeutic use , Cytomegalovirus Infections/immunology , Female , Glucocorticoids/therapeutic use , Herpesviridae Infections/diagnosis , Herpesviridae Infections/immunology , Herpesvirus 4, Human/immunology , Humans , Immunosuppressive Agents/therapeutic use , Lymphoproliferative Disorders/immunology , Male , Methylprednisolone/therapeutic use , Middle Aged , Postoperative Complications/immunology , Postoperative Complications/virology
20.
G Batteriol Virol Immunol ; 86(1-12): 29-42, 1994.
Article in Italian | MEDLINE | ID: mdl-8706973

ABSTRACT

HCMV infection is a major cause of morbidity and mortality following kidney transplantation. Clinical diagnosis is difficult, and rapid and sensitive diagnostic methods are needed since antiviral therapy is available. One hundred-forty-five consecutive kidney-transplanted patients were studied during a period of three months after transplantation. For laboratory diagnosis of HCMV infection, we looked for the presence of pp-65 antigen in polymorphonuclear leukocytes, HCMV-DNA and IgM. Demonstration of HCMV pp-65 antigen by immunofluorescence and HCMV DNA by PCR in leukocytes were efficient methods for early diagnosis of infection.


Subject(s)
Antigens, Viral/analysis , Cytomegalovirus Infections/diagnosis , Immunoglobulin G/analysis , Immunoglobulin M/analysis , Kidney Transplantation , Cytomegalovirus Infections/immunology , DNA, Viral/analysis , Humans , Immunocompromised Host
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