Changes in T cell effector functions over an 8-year period with TNF antagonists in patients with chronic inflammatory rheumatic diseases

The aim of the study was to clarify the effect of long-term anti-TNF therapy on T cell function in patients with rheumatologic immune-mediated inflammatory diseases (IMID). The production of IFNγ by T cells was evaluated at baseline and after 1, 2, 4, and 8 years of anti-TNF agents by means of a QuantiFERON-TB Gold In-Tube assay. The T cell proliferation and surface co-expression of CD25/CD134 in response to phytohaemagglutinin together with the in vitro impact of anti-TNF therapy on the functional capacity of T cells were evaluated after 8 years from the onset of the biological treatment. Age-matched healthy donors were enrolled as controls. The quantitative mitogen-induced IFNγ responses significantly increased with respect to baseline at each time point, apart from the determination after 4 years. We found an increased expression of CD25/CD134 in CD4+compared to CD8+T cells both in patients and controls. The in vitro addition of anti-TNF agents induced a significant decrease of both the IFNγ response and of CD25/CD134, whereas no effect on the intensity of the proliferative response was observed. Our data provide a biological basis for the reassuring issues on the safety of long-term anti-TNF treatment in patients with IMID

Comparison of the effects of air-powder abrasion, chemical decontamination, or their combination in open-flap surface decontamination of implants failed for peri-implantitis: an ex vivo study

Objectives To compare, using an ex vivo model, the biofilm removal of three surface decontamination methods following surgical exposure of implants failed for severe peri-implantitis. Materials and methods The study design was a single-blind, randomized, controlled, ex vivo investigation with intra-subject control. Study participants were 20 consecutive patients with at least 4 hopeless implants, in function for >12 months and with progressive bone loss exceeding 50%, which had to be explanted. Implants of each patient were randomly assigned to the untreated control group or one of the three decontamination procedures: mechanical debridement with air-powder abrasion, chemical decontamination with hydrogen peroxide and chlorhexidine gluconate, or combined mechanical-chemical decontamination. Following surgical exposure, implants selected as control were retrieved, and afterwards, test implants were decontaminated according to allocation and carefully explanted with a removal kit. Microbiological analysis was expressed in colony-forming-units (CFU/ml). Results A statistically significant difference (p < 0.001) in the concentrations of CFU/ml was found between implants treated with mechanical debridement (531.58 ± 372.07) or combined mechanical-chemical decontamination (954.05 ± 2219.31) and implants untreated (37,800.00 ± 46,837.05) or treated with chemical decontamination alone (29,650.00 ± 42,596.20). No statistically significant difference (p = 1.000) was found between mechanical debridement used alone or supplemented with chemical decontamination. Microbiological analyses identified 21 microbial species, without significant differences between control and treatment groups. Conclusions Bacterial biofilm removal from infected implant surfaces was significantly superior for mechanical debridement than chemical decontamination. Clinical relevance The present is the only ex vivo study based on decontamination methods for removing actual and mature biofilm from infected implant surfaces in patients with peri-implantitis

Dendritic cells in blood and urine samples from bladder cancer patients undergoing BCG immunotherapy

Objectives: Immunotherapy with BCG (Bacille Calmette-Guérin) after transurethral resection of the bladder tumor represents a highly effective primary treatment for intermediate and high-risk superficial bladder cancer. The effectiveness of this therapy has been documented, but its mechanism of action is not clear yet. In the present study, we investigated the changes of dendritic cells (DC) numbers in peripheral blood and urine of patients with superficial bladder cancer undergoing BCG intravescical therapy Material and method: We have enumerated plasmacytoid and myeloid DCs in the peripheral blood and in the urine of patients with bladder cancer in order to clarify the role of these cells in the evolution of the disease and the effect of therapy. DCs in blood and urine samples were assessed using the single-platform TruCOUNT assay with monoclonal antibodies. The study population included 37 healthy donors and 13 patients with diagnosis of primitive superficial bladder cancer. Results: At the time of diagnosis a reduction of blood DCs was found in patients as opposed to healthy donors, while DCs were not found in the urine in the same way as in healthy subjects. Six of these patients were followed before and after weekly and monthly instillations of BCG. In the peripheral blood, we observed an immunological recovery of DCs from the third weekly instillation up to the sixth. In the urine of patients, we didn't find mDCs or pDCs at T0, but we found a statistically significant change from the third instillation up to the sixth. On the contrary, we didn't find mDCs in urine during monthly instillation. Conclusions: DC Count could be used in the monitoring of patients undergoing BCG therapy. Immunological restoration of mDC numbers in peripheral blood and the efflux in urine could be important for confirming the effectiveness of BCG instillation

In Vivo and In Vitro Effects of Antituberculosis Treatment on Mycobacterial Interferon-γ T Cell Response

Background: In recent years, the impact of antituberculous treatment on interferon (IFN)-c response to Mycobacterium tuberculosis antigens has been widely investigated, but the results have been controversial. The objective of the present study was: i) to evaluate longitudinal changes of IFN-c response to M. tuberculosis-specific antigens in TB patients during antituberculous treatment by using the QuantiFERON-TB Gold (QFT-G) assay; ii) to compare the differences in T-cell response after a short or prolonged period of stimulation with mycobacterial antigens; iii) to assess the CD4+ and CD8+ T cells with effector/memory and central/memory phenotype; iv) to investigate the direct in vitro effects of antituberculous drugs on the secretion of IFN-c. Principal Findings: 38 TB patients was evaluated at baseline and at month 2 and 4 of treatment and at month 6 (treatment completion). 27 (71%) patients had a QFT-G reversion (positive to negative) at the end of therapy, while 11 (29%) TB patients remained QFT-G positive at the end of therapy. Among the 11 patients with persistent positive QFT-G results, six had a complete response to the treatment, while the remaining 5 patients did not have a resolution of the disease. All 27 patients who became QFT-G negative had a complete clinical and microbiological recovery of the TB disease. In these patients the release of IFN-c is absent even after a prolonged 6-day incubation with both ESAT-6 and CFP-10 antigens and the percentage of effector/memory T-cells phenotype was markedly lower than subjects with persistent positive QFT-G results. The in vitro study showed that antituberculous drugs did not exert any inhibitory effect on IFN-c production within the range of therapeutically achievable concentrations. Conclusions: The present study suggests that the decrease in the M. tuberculosis-specific T cells responses following successful anti-TB therapy may have a clinical value as a supplemental tool for the monitoring of the efficacy of pharmacologic intervention for active TB. In addition, the antituberculous drugs do not have any direct down-regulatory effect on the specific IFN-c response

Severe and Persistent Depletion of Circulating Plasmacytoid Dendritic Cells in Patients with 2009 Pandemic H1N1 Infection

Background: Dysregulation of host immune responses plays a critical role in the pathogenesis of severe 2009 pandemic H1N1 infection. Whether H1N1 virus could escape innate immune defense in vivo remains to be investigated. The aim of this study was to evaluate the pattern of innate immune response during human 2009 H1N1 infection. We performed the enumeration of circulating myeloid dendritic cells (mDC) and plasmacytoid DC (pDC) in blood from patients with H1N1 pneumonia shortly after the onset of symptoms and during follow-up at different intervals of time. The analysis of CD4 and CD8 count, CD38 T-cell activation marker and serum cytokine/chemokine plasma levels was also done. Methodology/Principal Findings: Blood samples were collected from 13 hospitalized patients with confirmed H1N1-related pneumonia at time of admission and at weeks 1, 4, and 16 of follow-up. 13 healthy donors were enrolled as controls. In the acute phase of the disease, H1N1-infected patients exhibited a significant depletion in both circulating pDC and mDC in conjunction with a decrease of CD4 and CD8 T cell count. In addition, we found plasmatic hyperproduction of IP-10 and RANTES, whereas increase in T-cell immune activation was found at all time points. When we assessed the changes in DC count over time, we observed a progressive normalization of mDC number. On the contrary, H1N1-infected patients did not achieve a complete recovery of pDC count as values remained lower than healthy controls even after 16 weeks of follow-up. Conclusions: H1N1 disease is associated with a profound depletion of DC subsets. The persistence of pDC deficit for several weeks after disease recovery could be due to H1N1 virus itself or to a preexisting impairment of innate immunity

Accuracy of Immunodiagnostic Tests for Active Tuberculosis Using Single and Combined Results: A Multicenter TBNET-Study

The clinical application of IFN-gamma release assays (IGRAs) has recently improved the diagnosis of latent tuberculosis infection. In a multicenter study of the Tuberculosis Network European Trialsgroup (TBNET) we aimed to ascertain in routine clinical practice the accuracy of a novel assay using selected peptides encoded in the mycobacterial genomic region of difference (RD) 1 for the diagnosis of active tuberculosis in comparison with tuberculin skin test (TST), QuantiFERON-TB GOLD In-Tube (Cellestis Ltd., Carnegie, Australia) and T-SPOT.TB (Oxfordimmunotec, Abingdon, UK)

Detecting latent tuberculosis in compromised patients

The detection of latent tuberculosis infection (LTBI) in different categories of compromised patients isreviewed with focus on the role of strategies incorporating immunodiagnostic tests and analysis ofepidemiological and clinical risk factors.Recent findingsThe development of active tuberculosis (TB) is increased in compromised patients and is closely related todeterminants for disease reactivation or newly acquired TB infection. A targeted detection of LTBI in thesehigh-risk groups should be performed especially if preventive treatment is planned. The performance ofimmunodiagnostic tests is highly variable among different groups of immunocompromised individuals.Findings of cross-sectional studies indicate a better diagnostic accuracy of interferon-grelease assays overthe tuberculin skin test. The critical issue is that in low-incidence countries, the positive and negativepredictive values of any of immunodiagnostic tests were very poor. A targeted testing process involvinganalysis of TB risk factors increases the predictive positive values of immunodiagnostic tests and mayimprove LTBI detection.SummaryThe LTBI detection in immunocompromised patients is a challenge. The development of new immunologicalbiomarkers and integrated clinical and epidemiological strategies are needed to identify LTBI incompromised individuals and to plan preventive chemotherapies in those at risk of developing active TB

Rapid identification of mycobacterial species directly in clinical samples

Up to today, the prompt diagnosis of NTM infection has been impaired by the slow growth in culture media, which is an essential step for proper identification. The aim of our study was to evaluate the kit Genotype Mycobacterium CM/AS for the identification of NTM in clinical specimens. Four patients admitted to the Department of Infectious and Tropical Diseases of Sapienza University in Rome were included in the study.Three out of 4 patients were strongly suspected to have a NTM infection and one of them underwent treatment for M. tuberculosis infection without any substantial clinical improvement. After decontamination of the specimens, the extracted DNA was amplified in target sequences using PCR assay by increasing the number of cycles from 20 to 30. Subsequently the Genotype assay was performed according to the manufacturer’s instructions. Our results confirmed the presence of NTM in all patients: M. peregrinum in two patients, M. gordone and M. intracellulare in the others. The use of the Genotype Mycobacterium CM/AS directly from clinical specimens permits rapid diagnosis and enables clinicians to start an effective treatment