RESUMO
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.
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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.
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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.
Assuntos
Elétrons , Compostos Organometálicos/química , Processos Fotoquímicos , Estrutura Molecular , Teoria QuânticaRESUMO
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.
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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.
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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.
Assuntos
DNA Viral/genética , Vírus JC/genética , Reação em Cadeia da Polimerase/métodos , Sequência de Bases , Estudos de Casos e Controles , DNA Viral/análise , DNA Viral/líquido cefalorraquidiano , Humanos , Vírus JC/isolamento & purificação , Transplante de Rim , Leucoencefalopatia Multifocal Progressiva/virologia , Reação em Cadeia da Polimerase/normasRESUMO
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.
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Antígenos Virais/sangue , Citomegalovirus/isolamento & purificação , Fosfoproteínas/sangue , Fosfoproteínas/genética , Reação em Cadeia da Polimerase/métodos , RNA Mensageiro/análise , Proteínas da Matriz Viral/sangue , Proteínas da Matriz Viral/genética , Viremia/diagnóstico , Adulto , Idoso , DNA Viral/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico , Carga ViralRESUMO
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.
Assuntos
Apolipoproteínas/farmacologia , Candida albicans/imunologia , Degranulação Celular/efeitos dos fármacos , Ativação de Neutrófilo , Neutrófilos/imunologia , Fagocitose/efeitos dos fármacos , Proteína Amiloide A Sérica/farmacologia , Reação de Fase Aguda , Adulto , Antígenos CD/metabolismo , Cálcio/metabolismo , Candida albicans/citologia , Candida albicans/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Humanos , Lactoferrina/metabolismo , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Neutrófilos/microbiologia , Proteínas Recombinantes/farmacologia , Explosão Respiratória/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Fatores de Virulência de Bordetella/farmacologiaRESUMO
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.
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Cicatriz Hipertrófica/genética , Interleucina-15/genética , Interleucina-15/fisiologia , Adolescente , Adulto , Idoso , Apoptose/efeitos dos fármacos , Complexo CD3 , Ciclo Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Cicatriz Hipertrófica/metabolismo , Cicatriz Hipertrófica/patologia , Feminino , Expressão Gênica , Humanos , Imuno-Histoquímica , Interleucina-15/farmacologia , Interleucina-2/farmacologia , Contagem de Linfócitos , Masculino , Pessoa de Meia-Idade , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/efeitos dos fármacos , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Linfócitos T/citologia , Linfócitos T/imunologiaRESUMO
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.
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Anemia/etiologia , Infecções por Citomegalovirus/virologia , DNA Viral/sangue , Transplante de Rim , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/isolamento & purificação , Complicações Pós-Operatórias/virologia , Proteínas Recombinantes de Fusão , Viremia/virologia , Anemia/virologia , Anticorpos Monoclonais/efeitos adversos , Anticorpos Antivirais/sangue , Soro Antilinfocitário/efeitos adversos , Basiliximab , Ciclosporina/efeitos adversos , Infecções por Citomegalovirus/etiologia , Infecções por Citomegalovirus/terapia , DNA Viral/isolamento & purificação , Diagnóstico Diferencial , Suscetibilidade a Doenças , Feminino , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Imunossupressores/efeitos adversos , Interleucina-1/antagonistas & inibidores , Masculino , Ácido Micofenólico/efeitos adversos , Ácido Micofenólico/análogos & derivados , Infecções por Parvoviridae/etiologia , Infecções por Parvoviridae/terapia , Parvovirus B19 Humano/genética , Parvovirus B19 Humano/imunologia , Fosfoproteínas/sangue , Reação em Cadeia da Polimerase , Prednisona/efeitos adversos , Estudos Retrospectivos , Linfócitos T , Tacrolimo/efeitos adversos , Carga Viral , Proteínas da Matriz Viral/sangue , Zidovudina/efeitos adversosRESUMO
Anti-HIV antibodies were detected in postmortem blood and vitreous humor samples from 60 drug addicts died in 1988 and medicolegally autopsied at the University Institute of Forensic Science of Turin. Fifteen subjects were positive both in blood and in vitreous samples confirming the possibility to detect anti-HIV antibodies in vitreous humor in the screening of high-risk population.
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Autopsia/métodos , Anticorpos Anti-HIV/análise , Corpo Vítreo/imunologia , Sangue/imunologia , HumanosRESUMO
Anti-HTLV-I antibody presence in drug addicts dead in 1987-1988 in Turin, was evaluated. Comparing the prevalence of HTLV-I infection with that of HIV and HBV infections, a different way and/or rate of spread of HTLV-I infection is suggested.
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Infecções por HTLV-I/epidemiologia , Transtornos Relacionados ao Uso de Substâncias/complicações , Feminino , Infecções por HIV/complicações , Infecções por HIV/epidemiologia , Infecções por HTLV-I/complicações , Hepatite B/complicações , Hepatite B/epidemiologia , Humanos , Itália/epidemiologia , MasculinoRESUMO
Serological markers of HIV and HBV infections were studied in 90 drug addicts who died in 1988 and were medicolegally autopsied at the Institute of Forensic Sciences, University of Turin. Nineteen (21.1%) displayed evidence of HIV infection, demonstrated by the presence of anti-HIV antibodies; fifty-nine (65.5%) HBV infection, demonstrated by the presence of anti-HBc antibodies and/or HBsAg; nine (10%) had HBsAg, indicating potential infectiousness for HBV infection.
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Anticorpos Anti-HIV/análise , Anticorpos Anti-Hepatite B/análise , Antígenos de Superfície da Hepatite B/análise , Transtornos Relacionados ao Uso de Substâncias/imunologia , Adolescente , Adulto , Feminino , Humanos , Itália , Masculino , Pessoa de Meia-Idade , Transtornos Relacionados ao Uso de Substâncias/mortalidadeRESUMO
CMV-specific IgA, IgM and IgG antibodies were detected by ELISA in sera from 81 renal transplant patients. Twenty-seven patients were followed from transplantation; 6 patients who underwent on transplantation before the beginning of the study were followed during admissions for graft failure or acute illness; 48 outpatients were periodically monitored. One of the patient followed from transplantation experienced a primary CMV infection, serologically demonstrated by the appearance of specific IgM and IgG. Specific IgA appeared at the same time as IgM and lasted for about six months. A specific IgA response was observed in all but five recurrent CMV infections too, even when specific IgM were not present. In all outpatients periodically monitored for CMV serology specific IgA were not found. About specific IgA polimerization, a transient marked polymeric IgA (p-IgA) response was observed in only the primary infection whereas in all the other IgA positive patients, specific IgA were represented by monomers (m-IgA).
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Anticorpos Antivirais/análise , Citomegalovirus/imunologia , Imunoglobulina A/análise , Transplante de Rim , Adulto , Humanos , Imunoglobulina M/análise , Pessoa de Meia-IdadeRESUMO
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.
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Vírus BK/genética , DNA Viral/sangue , DNA Viral/urina , Vírus JC/genética , Transplante de Rim , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Infecções por Polyomavirus/epidemiologia , Sensibilidade e Especificidade , Infecções Tumorais por Vírus/epidemiologiaRESUMO
The relationship between immunoglobulin abnormalities and EBV infection has been investigated in 65 renal transplant patients. Immunoglobulins abnormalities were demonstrated in 44 (68%) patients, 8 of them (18%, 12% of all patients) had a monoclonal component. Up to now no lymphoproliferative disorder has been observed in these patients. All patients were EBV seropositive at the beginning of the study and in 22 of them (33%) a reactivation of EBV infection could be demonstrated. No relation has been observed between immunoglobulin abnormalities, EBV reactivation, age or sex. By contrast, a significant relation was found between EBV reactivation and immunosuppressive treatment: patients under triple therapy with Azathioprine, Cyclosporine A and Prednisone had less EBV reactivation compared to those under Cyclosporine A treatment.
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Infecções por Herpesviridae/sangue , Herpesvirus Humano 4 , Cadeias kappa de Imunoglobulina/sangue , Cadeias lambda de Imunoglobulina/sangue , Transplante de Rim , Gamopatia Monoclonal de Significância Indeterminada/microbiologia , Complicações Pós-Operatórias/microbiologia , Adolescente , Adulto , Ciclosporina/efeitos adversos , Suscetibilidade a Doenças , Feminino , Infecções por Herpesviridae/complicações , Infecções por Herpesviridae/imunologia , Herpesvirus Humano 4/fisiologia , Humanos , Hospedeiro Imunocomprometido , Imunossupressores/efeitos adversos , Falência Renal Crônica/complicações , Falência Renal Crônica/cirurgia , Transtornos Linfoproliferativos/etiologia , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/sangue , Ativação ViralRESUMO
Human herpes virus type 6 (HHV-6) infection was serologically investigated in renal transplant recipients. Before transplantation, 75.5% of patients was seropositive for HHV-6 and no correlation with age, sex and time on dialysis was found. During the first month after transplantation 66% of patients showed a variation in serological status against HHV-6 (seroconversion or fourfold increase of antibody titer). All patients who seroconverted had received the kidney from a HHV-6 seropositive donor, furthermore, in 11/13 (84.6%) pairs of patients receiving the kidney form the same seropositive donor, both members or had HHV-6 active infection or had no infection. The frequency of HHV-6 active infection in seropositive patients is almost the same in case of seronegative or seropositive donor. Comparing HHV-6 and CMV infections, they resulted independent as CMV infection in these patients occurs in a following period (II-III month). Notwithstanding a higher frequency of kidney rejection in patients with active HHV-6 infection, no significative correlation was found.
Assuntos
Infecções por Herpesviridae/epidemiologia , Herpesvirus Humano 6/isolamento & purificação , Transplante de Rim , Complicações Pós-Operatórias/microbiologia , Adulto , Anticorpos Antivirais/biossíntese , Criança , Infecções por Citomegalovirus/complicações , Infecções por Citomegalovirus/epidemiologia , Suscetibilidade a Doenças , Feminino , Rejeição de Enxerto , Infecções por Herpesviridae/complicações , Infecções por Herpesviridae/transmissão , Herpesvirus Humano 6/imunologia , Humanos , Hospedeiro Imunocomprometido , Imunossupressores/efeitos adversos , Falência Renal Crônica/complicações , Falência Renal Crônica/terapia , Transplante de Rim/efeitos adversos , Masculino , Pessoa de Meia-Idade , Prevalência , Diálise Renal , Ativação ViralRESUMO
Variations in anti-CMV antibody affinity have been studied in 106 renal transplant patients. Maturation of immune response has been followed during two years after transplantation evaluating antibody affinity by ELISA before and after urea denaturation treatment. In primary infections while the antibody titer rises, the resistance to denaturing treatment rises as well, indicating an increased antibody affinity. In patients already seropositive at transplantation, the increase of antibody affinity has also been found: comparing the affinity index (AI) at transplantation and one year later, only 16.5% of patients showed an AI value between 80 and 100 at transplantation, whereas after one year 62.3% of patients reached such AI values (p = 0.0001).
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Anticorpos Antivirais/imunologia , Citomegalovirus/imunologia , Transplante de Rim , Anticorpos Antivirais/biossíntese , Afinidade de Anticorpos , Complexo Antígeno-Anticorpo/efeitos dos fármacos , Infecções por Citomegalovirus/imunologia , Ensaio de Imunoadsorção Enzimática , Seguimentos , Humanos , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Complicações Pós-Operatórias/imunologia , Desnaturação Proteica , Ureia/farmacologiaRESUMO
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.
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Infecções por Citomegalovirus/terapia , Infecções por Herpesviridae/terapia , Herpesvirus Humano 4/isolamento & purificação , Transplante de Rim , Transtornos Linfoproliferativos/terapia , Adulto , Anticorpos Antivirais/imunologia , Ciclosporina/uso terapêutico , Infecções por Citomegalovirus/imunologia , Feminino , Glucocorticoides/uso terapêutico , Infecções por Herpesviridae/diagnóstico , Infecções por Herpesviridae/imunologia , Herpesvirus Humano 4/imunologia , Humanos , Imunossupressores/uso terapêutico , Transtornos Linfoproliferativos/imunologia , Masculino , Metilprednisolona/uso terapêutico , Pessoa de Meia-Idade , Complicações Pós-Operatórias/imunologia , Complicações Pós-Operatórias/virologiaRESUMO
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.