Alessandro Sanduzzi | Università degli Studi di Napoli "Federico II" (original) (raw)

Papers by Alessandro Sanduzzi

Respiratory Medicine, 2007

Expert Opinion on Drug Metabolism & Toxicology, 2015

Understanding variability in the response to asthma medications is essential to ensure appropriat... more Understanding variability in the response to asthma medications is essential to ensure appropriate prescribing. Given that there are increased asthma treatment failures observed in ethnic minorities receiving asthma therapeutics, it is fundamental to understand the factors related to ethnicity that can modify the response to asthma therapy. Race/ethnicity is an important determinant of drug response and therefore contributes to interindividual variability. It is generally recognized that its effects on drug response are determined by both genetic and environmental factors to a varying extent, depending on the ethnic groups and probe drugs studied. Also, adherence to therapy can influence pharmacological response to asthma therapeutics. Health-care professionals might never use the treatment in their patients irrespective of their ethnicity and thus inadvertently increase ethnic health inequality. However, our understanding of whether and/or how ethnicity influences pharmacological response to asthma therapeutics is still very scarce. A holistic, integrative systems biology approach that combines large-scale molecular profiling traits (e.g., transcriptomic, proteomic, metabolomic traits) and genetic variants could help to personalize the treatment of asthmatic patients regardless of race/ethnicity.

Le infezioni in medicina : rivista periodica di eziologia, epidemiologia, diagnostica, clinica e terapia delle patologie infettive, 2014

In recent years Tumor Necrosis Factor alpha (TNF alfa) inhibitors have been highly effective in t... more In recent years Tumor Necrosis Factor alpha (TNF alfa) inhibitors have been highly effective in treating rheumatoid arthritis (RA). However, patients receiving these inhibitors have an increased risk of developing tuberculosis (TB). We describe a rare case of tuberculosis of the tongue in an RA patient treated with methotrexate (MTX) and the TNF alfa inhibitor adalimumab (ADA) for the previous six years. Pretreatment tuberculin skin test (TST) was negative. The patient was admitted to our division complaining of a sore throat for months. Clinical examination revealed a swollen non-healing ulcer at the base of the tongue, which was suspected to be a squamous cell carcinoma. Histopathological assessment unexpectedly revealed a chronic granulomatous inflammation compatible with tuberculosis. TST was strongly positive and the T Spot TB test was also reactive. MTX and ADA were discontinued and the patient received antituberculous treatment with complete healing of the lesion. After three...

Pulmonary pharmacology & therapeutics, 2003

Emergence of antimicrobial resistance is a growing problem and a public health threat. New drugs ... more Emergence of antimicrobial resistance is a growing problem and a public health threat. New drugs must be designed with emerging needs in mind: specific resistant and hard-to-treat organisms. But the difficulty to find real new drugs is a major problem. Only the oxazolidinones, the cationic peptides and the lipopeptide antibiotics can be truly regarded as structurally novel drugs, although the peptide deformylase inhibitors and, possibly, the pleuromutilins can be considered a potential advancement in the field. Obviously, these antibiotics must be reserved only to cases of documented ineffectiveness of the common antimicrobial agents.

Pulmonary Pharmacology & Therapeutics, 2015

Inhalation is the preferred route of drug administration in chronic respiratory diseases because ... more Inhalation is the preferred route of drug administration in chronic respiratory diseases because it optimises delivery of the active compounds to the targeted site and minimises side effects from systemic distribution. The choice of a device should be made after careful evaluation of the patient's clinical condition (degree of airway obstruction, comorbidities), as well as their ability to coordinate the inhalation manoeuvre and to generate sufficient inspiratory flow. These patient factors must be aligned with the specific advantages and limitations of each inhaler when making this important choice. Finally, adherence to treatment is not the responsibility of the patient alone, but should be shared also by clinicians. Clinicians have access to a wide selection of pressurised metered dose inhalers (pMDIs) and dry powder inhalers (DPIs) that can be used effectively when matched to the needs of individual patients; this should be perceived as an opportunity rather than a limitation.

Multidisciplinary Respiratory Medicine, 2014

Increased asthma severity is not only associated with enhanced recurrent hospitalization and mort... more Increased asthma severity is not only associated with enhanced recurrent hospitalization and mortality but also with higher social costs. Several cases of asthma are atopic in nature, with the trigger for acute asthma attacks and chronic worsening of inflammation being allergens inducing an immune, IgE mediated response. Anti-inflammatory treatments are effective for most of asthma patients, but there are subjects whose disease is incompletely controlled by inhaled or systemic corticosteroids and these patients account for about 50% of the healthcare costs of asthma. Omalizumab is a biological engineered, humanized recombinant monoclonal anti-IgE antibody developed for the treatment of allergic diseases and with clear efficacy in adolescent and adult patients with severe allergic asthma. The anti-IgE antibody inhibits IgE functions blocking free serum IgE and inhibiting their binding to cellular receptors. By reducing serum IgE levels and IgE receptor expression on inflammatory cells in the context of allergic cascade, omalizumab has demonstrated to be a very useful treatment of atopic asthma, improving quality of life of patients with severe persistent allergic asthma that is inadequately controlled by currently available asthma medications. Several trials have demonstrated that this therapy is well tolerated and significantly improves symptoms and disease control, reducing asthma exacerbations and the need to use high dosage of inhaled corticosteroids.

Autoimmunity Reviews, 2015

on behalf of SAFEBIO (Italian multidisciplinary task force for screening of tuberculosis before a... more on behalf of SAFEBIO (Italian multidisciplinary task force for screening of tuberculosis before and during biologic therapy) a b s t r a c t a r t i c l e i n f o Available online xxxx

Respiratory Medicine, 2007

Background: Induced sputum (IS) sampling is a safe and validated approach to study bronchial infl... more Background: Induced sputum (IS) sampling is a safe and validated approach to study bronchial inflammation in chronic obstructive lung diseases. Although promising results have also been reported in various diffuse interstitial lung disorders, the potential use of IS in the assessment of connective tissue diseases (CTD)-related lung involvement has not yet been investigated. Aim of the study: To evaluate the clinical usefulness of IS in the early management of patients suffering from rheumatoid arthritis (RA) and systemic sclerosis (SSc) at the onset of respiratory symptoms. Patients and methods: The study population included 19 patients (RA ¼ 12; SSc ¼ 7) and 14 age-and sex-matched healthy volunteers. Lung function testing, high resolution computed tomography (HRCT) of the thorax and IS collection were performed in all cases. Broncho-alveolar lavage (BAL) was obtained in selected patients. Results: IS samples from patients contained a significantly higher percentage of neutrophils and a lower percentage of macrophages compared to healthy subjects (p ¼ 0.002 and 0.001, respectively), while the total cell number showed no differences. In addition, sputa yielded both higher cell counts and higher neutrophils than BAL samples (p ¼ 0.02 in all instances). No correlations were found between IS findings and lung function parameters, HRCT and BAL findings. (A. Vatrella).

Respiratory Medicine, 2010

M. Bocchino). a v a i l a b l e a t w w w . s c i e n c e d i r e c t . c o m j o u r n a l h o m... more M. Bocchino). a v a i l a b l e a t w w w . s c i e n c e d i r e c t . c o m j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / r m e d Respiratory Medicine (2010) 104, 1551e1556 0954-6111/$ -see front matter ª

PLoS ONE, 2009

CD8 T-cells contribute to control of Mycobacterium tuberculosis infection, but little is known ab... more CD8 T-cells contribute to control of Mycobacterium tuberculosis infection, but little is known about the quality of the CD8 Tcell response in subjects with latent infection and in patients with active tuberculosis disease. CD8 T-cells recognizing epitopes from 6 different proteins of Mycobacterium tuberculosis were detected by tetramer staining. Intracellular cytokines staining for specific production of IFN-c and IL-2 was performed, complemented by phenotyping of memory markers on antigen-specific CD8 T-cells. The ex-vivo frequencies of tetramer-specific CD8 T-cells in tuberculous patients before therapy were lower than in subjects with latent infection, but increased at four months after therapy to comparable percentages detected in subjects with latent infection. The majority of CD8 T-cells from subjects with latent infection expressed a terminally-differentiated phenotype (CD45RA + CCR7 2 ). In contrast, tuberculous patients had only 35% of antigen-specific CD8 T-cells expressing this phenotype, while containing higher proportions of cells with an effector memory-and a central memory-like phenotype, and which did not change significantly after therapy. CD8 T-cells from subjects with latent infection showed a codominance of IL-2 + /IFN-c + and IL-2 2 /IFN-c + T-cell populations; interestingly, only the IL-2 + /IFN-c + population was reduced or absent in tuberculous patients, highly suggestive of a restricted functional profile of Mycobacterium tuberculosis-specific CD8 T-cells during active disease. These results suggest distinct Mycobacterium tuberculosis specific CD8 T-cell phenotypic and functional signatures between subjects which control infection (subjects with latent infection) and those who do not (patients with active disease).

Otolaryngology - Head and Neck Surgery, 2003

The Journal of Pathology, 1997

Tuberculosis (TB) is still a major health problem, both as a single disease entity and as a cofac... more Tuberculosis (TB) is still a major health problem, both as a single disease entity and as a cofactor in AIDS. The interaction between macrophage and Mycobacterium tuberculosis (MTB) is a critical step in the establishment of an early chronic infection. This study analyses the capacity of MTB to induce apoptosis in cells obtained by broncho-alveolar lavage (BAL) from patients with reactive pulmonary tuberculosis and from AIDS patients with disseminated pulmonary tuberculosis. Apoptosis was increased three-fold in BAL cells obtained from patients with pulmonary tuberculosis and even more markedly in alveolar macrophages of MTB-infected AIDS patients, compared with controls. Apoptosis was analysed and characterized by propidium iodide (PI) incorporation, terminal deoxy transferase (TDT)-mediated dUTP-biotin nick end labelling (TUNEL), and tissue transglutaminase (tTG) expression. The MTB-macrophage interaction was also investigated in vitro by infecting monocyte-derived macrophages (MDM) with MTB (virulent strain H37Rv). The induction of apoptosis by MTB required viable bacteria, was dose-dependent, and was restricted to H37Rv. Infection with either Mycobacterium avium complex (MAC) or HIV-1 and treatment with heat-killed MTB failed to induce apoptosis.

Infection and Immunity, 2001

Mycobacterium tuberculosis is an intracellular pathogen that readily survives and replicates in h... more Mycobacterium tuberculosis is an intracellular pathogen that readily survives and replicates in human macrophages (M⌽). Host cells have developed different mycobactericidal mechanisms, including the production of inflammatory cytokines. The aim of this study was to compare the M⌽ response, in terms of cytokine gene expression, to infection with the M. tuberculosis laboratory strain H37Rv and the clinical M. tuberculosis isolate CMT97. Both strains induce the production of interleukin-12 (IL-12) and IL-16 at comparable levels. However, the clinical isolate induces a significantly higher and more prolonged M⌽ activation, as shown by reverse transcription-PCR analysis of IL-1␤, IL-6, IL-10, transforming growth factor beta, tumor necrosis factor alpha, and gamma interferon (IFN-␥) transcripts. Interestingly, when IFN-␥ transcription is high, the number of M. tuberculosis genes expressed decreases and vice versa, whereas no mycobactericidal effect was observed in terms of bacterial growth. Expression of 11 genes was also studied in the two M. tuberculosis strains by infecting resting or activated M⌽ and compared to bacterial intracellular survival. In both cases, a peculiar inverse correlation between expression of these genes and multiplication was observed. The number and type of genes expressed by the two strains differed significantly.

Immunology, 2004

This study addresses the analysis of the human leucocyte antigen (HLA) allele distribution in 54 ... more This study addresses the analysis of the human leucocyte antigen (HLA) allele distribution in 54 historical and in 68 recently diagnosed tuberculosis (TB) patients. The historical cohort was characterized by the presence of large ®brocavernous lesions effectively treated with therapeutic pneumothorax during the period 1950±55. Patients and healthy controls enrolled in the study were from the Campania region of southern Italy. No signi®cant association between HLA alleles and TB in the population of recently diagnosed TB patients was observed. On the contrary, among the historical TB patients there was a strong association with an increased frequency of the HLA-DR4 allele alone and/or in the presence of the HLA-B14 allele (P 0Á000004; P c 0Á0008), as well as with a decreased frequency of the HLA-A2 ,-B14 À ,DR4 À allele association (P 0Á00005; P c 0Á01). In order to exclude any interference from age-related factors, these results were con®rmed by comparing the historical cohort of TB patients with an age-matched healthy control population of the same ethnic origin (P 0Á00004; P c 0Á008; and P 0Á0001; and P c 0Á02, respectively).

European Respiratory Journal, 2003

European Journal of Immunology, 2010

Th1 CD4 1 T cells and their derived cytokines are crucial for protection against Mycobacterium tu... more Th1 CD4 1 T cells and their derived cytokines are crucial for protection against Mycobacterium tuberculosis. Using multiparametric flow cytometry, we have evaluated the distribution of seven distinct functional states (IFN-c/IL-2/TNF-a triple expressors, IFN-c/IL-2, IFN-c/TNF-a or TNF-a/IL-2 double expressors or IFN-c, IL-2 or TNF-a single expressors) of CD4 1 T cells in individuals with latent M. tuberculosis infection (LTBI) and active tuberculosis (TB). We found that triple expressors, while detectable in 85-90%TB patients, were only present in 10-15% of LTBI subjects. On the contrary, LTBI subjects had significantly higher (12-to 15-fold) proportions of IL-2/IFN-c double and IFN-c single expressors as compared with the other CD4 1 T-cell subsets. Proportions of the other double or single CD4 1 T-cell expressors did not differ between TB and LTBI subjects. These distinct IFN-c, IL-2 and TNF-a profiles of M. tuberculosis-specific CD4 1 T cells seem to be associated with live bacterial loads, as indicated by the decrease in frequency of multifunctional T cells in TB-infected patients after completion of anti-mycobacterial therapy. Our results suggest that phenotypic and functional signatures of CD4 1 T cells may serve as immunological correlates of protection and curative host responses, and be a useful tool to monitor the efficacy of anti-mycobacterial therapy.

European Journal of Clinical Microbiology & Infectious Diseases, 2008

The reactivation of latent tuberculosis (TB) is a major complication of tumor necrosis factor (TN... more The reactivation of latent tuberculosis (TB) is a major complication of tumor necrosis factor (TNF)-alpha inhibitors. Screening for TB infection is recommended before anti-TNF therapy is initiated; however, the use of tuberculin skin testing (TST) is controversial, due to the high rate of false-negative results in patients receiving immunosuppressive treatment. To compare the performance of two commercial interferon (IFN)-gamma release assays (IGRA), T-SPOT.TB (TS-TB) and QuantiFERON-TB Gold "In-tube" (QFT-GIT), with TST for the detection of TB infection in patients due to start anti-TNF therapy, 69 human immunodeficiency virus (HIV)-negative Italian patients (mean age: 45.2 +/- 12.6 years; male=39) were enrolled between September 2005 to August 2006. Patients affected by rheumatoid arthritis (n = 18), psoriatic arthritis (n = 26), ulcerous rectocolitis (n = 6), and Crohn's disease (n = 19) were tested simultaneously with TST, TS-TB, and QFT-GIT. Overall, 26% of patients were positive by TST, 30.4% by TS-TB, and 31.8% by QFT-GIT. Agreement with TST was similar (kappa = 0.21, p = 0.0002 and kappa = 0.26, p < 0.001, respectively). In 11 TST-negative cases, IFN-gamma release assays were positive. In addition, in seven Mantoux-positive cases with no TB risk factors, TST result agreement was achieved with at least one blood test. Indeterminate results were detected in 5.8% and 2.8% of cases, respectively, with TS-TB and with QFT-GIT (p = not significant [ns]). In conclusion, our results suggest that IGRAs may be helpful for screening purposes in patient candidates for anti-TNF therapy to confirm positive TST results and in selected cases when false-negative results are suspected. The utility of blood tests in patients with low or no TB risk remains to be assessed.

Cytokine, 2014

The immune response plays an unsettled role in the pathogenesis of idiopathic pulmonary fibrosis ... more The immune response plays an unsettled role in the pathogenesis of idiopathic pulmonary fibrosis (IPF), the contribution of inflammation being controversial as well. Emerging novel T cell sub-populations including regulatory T lymphocytes (Treg) and interleukin (IL)-17 secreting T helper cells (Th17) may exert antithetical actions in this scenario. Phenotype and frequency of circulating immune cell subsets were assessed by multi-parametric flow cytometry in 29 clinically stable IPF patients and 17 healthy controls. The interplay between Treg lymphocytes expressing transforming growth factor (TGF)-β and Th17 cells was also investigated. Proportion and absolute number of natural killer (NK) cells were significantly reduced in IPF patients in comparison with controls (p<0.001). Conversely, the proportion and absolute number of CD3(+)CD4(+)CD25(high)Foxp-3(+) cells were significantly increased in IPF patients (p=0.000). As in controls, almost the totality of cells (>90%) expressed TGF-β upon stimulation. Interestingly, the frequency of Th17 cells was significantly compromised in IPF patients (p=0.000) leading to an increased TGF-β/IL-17 ratio (4.2±2.3 vs 0.5±0.3 in controls, p=0.000). Depletion of NK and Th17 cells along with a not compromised Treg compartment delineate the existence of an "immune profile" that argue against the recent hypothesis of IPF as an autoimmune disease. Our findings along with the imbalance of the Treg/Th17 axis more closely suggest these immune perturbations to be similar to those observed in cancer. Clinical relevance, limitations and perspectives for future research are discussed.

Clinical Rheumatology, 2014

Clinical Rheumatology, 2010

The aim of this study

Respiratory Medicine, 2007

Expert Opinion on Drug Metabolism & Toxicology, 2015

Understanding variability in the response to asthma medications is essential to ensure appropriat... more Understanding variability in the response to asthma medications is essential to ensure appropriate prescribing. Given that there are increased asthma treatment failures observed in ethnic minorities receiving asthma therapeutics, it is fundamental to understand the factors related to ethnicity that can modify the response to asthma therapy. Race/ethnicity is an important determinant of drug response and therefore contributes to interindividual variability. It is generally recognized that its effects on drug response are determined by both genetic and environmental factors to a varying extent, depending on the ethnic groups and probe drugs studied. Also, adherence to therapy can influence pharmacological response to asthma therapeutics. Health-care professionals might never use the treatment in their patients irrespective of their ethnicity and thus inadvertently increase ethnic health inequality. However, our understanding of whether and/or how ethnicity influences pharmacological response to asthma therapeutics is still very scarce. A holistic, integrative systems biology approach that combines large-scale molecular profiling traits (e.g., transcriptomic, proteomic, metabolomic traits) and genetic variants could help to personalize the treatment of asthmatic patients regardless of race/ethnicity.

Le infezioni in medicina : rivista periodica di eziologia, epidemiologia, diagnostica, clinica e terapia delle patologie infettive, 2014

In recent years Tumor Necrosis Factor alpha (TNF alfa) inhibitors have been highly effective in t... more In recent years Tumor Necrosis Factor alpha (TNF alfa) inhibitors have been highly effective in treating rheumatoid arthritis (RA). However, patients receiving these inhibitors have an increased risk of developing tuberculosis (TB). We describe a rare case of tuberculosis of the tongue in an RA patient treated with methotrexate (MTX) and the TNF alfa inhibitor adalimumab (ADA) for the previous six years. Pretreatment tuberculin skin test (TST) was negative. The patient was admitted to our division complaining of a sore throat for months. Clinical examination revealed a swollen non-healing ulcer at the base of the tongue, which was suspected to be a squamous cell carcinoma. Histopathological assessment unexpectedly revealed a chronic granulomatous inflammation compatible with tuberculosis. TST was strongly positive and the T Spot TB test was also reactive. MTX and ADA were discontinued and the patient received antituberculous treatment with complete healing of the lesion. After three...

Pulmonary pharmacology & therapeutics, 2003

Emergence of antimicrobial resistance is a growing problem and a public health threat. New drugs ... more Emergence of antimicrobial resistance is a growing problem and a public health threat. New drugs must be designed with emerging needs in mind: specific resistant and hard-to-treat organisms. But the difficulty to find real new drugs is a major problem. Only the oxazolidinones, the cationic peptides and the lipopeptide antibiotics can be truly regarded as structurally novel drugs, although the peptide deformylase inhibitors and, possibly, the pleuromutilins can be considered a potential advancement in the field. Obviously, these antibiotics must be reserved only to cases of documented ineffectiveness of the common antimicrobial agents.

Pulmonary Pharmacology & Therapeutics, 2015

Inhalation is the preferred route of drug administration in chronic respiratory diseases because ... more Inhalation is the preferred route of drug administration in chronic respiratory diseases because it optimises delivery of the active compounds to the targeted site and minimises side effects from systemic distribution. The choice of a device should be made after careful evaluation of the patient's clinical condition (degree of airway obstruction, comorbidities), as well as their ability to coordinate the inhalation manoeuvre and to generate sufficient inspiratory flow. These patient factors must be aligned with the specific advantages and limitations of each inhaler when making this important choice. Finally, adherence to treatment is not the responsibility of the patient alone, but should be shared also by clinicians. Clinicians have access to a wide selection of pressurised metered dose inhalers (pMDIs) and dry powder inhalers (DPIs) that can be used effectively when matched to the needs of individual patients; this should be perceived as an opportunity rather than a limitation.

Multidisciplinary Respiratory Medicine, 2014

Increased asthma severity is not only associated with enhanced recurrent hospitalization and mort... more Increased asthma severity is not only associated with enhanced recurrent hospitalization and mortality but also with higher social costs. Several cases of asthma are atopic in nature, with the trigger for acute asthma attacks and chronic worsening of inflammation being allergens inducing an immune, IgE mediated response. Anti-inflammatory treatments are effective for most of asthma patients, but there are subjects whose disease is incompletely controlled by inhaled or systemic corticosteroids and these patients account for about 50% of the healthcare costs of asthma. Omalizumab is a biological engineered, humanized recombinant monoclonal anti-IgE antibody developed for the treatment of allergic diseases and with clear efficacy in adolescent and adult patients with severe allergic asthma. The anti-IgE antibody inhibits IgE functions blocking free serum IgE and inhibiting their binding to cellular receptors. By reducing serum IgE levels and IgE receptor expression on inflammatory cells in the context of allergic cascade, omalizumab has demonstrated to be a very useful treatment of atopic asthma, improving quality of life of patients with severe persistent allergic asthma that is inadequately controlled by currently available asthma medications. Several trials have demonstrated that this therapy is well tolerated and significantly improves symptoms and disease control, reducing asthma exacerbations and the need to use high dosage of inhaled corticosteroids.

Autoimmunity Reviews, 2015

on behalf of SAFEBIO (Italian multidisciplinary task force for screening of tuberculosis before a... more on behalf of SAFEBIO (Italian multidisciplinary task force for screening of tuberculosis before and during biologic therapy) a b s t r a c t a r t i c l e i n f o Available online xxxx

Respiratory Medicine, 2007

Background: Induced sputum (IS) sampling is a safe and validated approach to study bronchial infl... more Background: Induced sputum (IS) sampling is a safe and validated approach to study bronchial inflammation in chronic obstructive lung diseases. Although promising results have also been reported in various diffuse interstitial lung disorders, the potential use of IS in the assessment of connective tissue diseases (CTD)-related lung involvement has not yet been investigated. Aim of the study: To evaluate the clinical usefulness of IS in the early management of patients suffering from rheumatoid arthritis (RA) and systemic sclerosis (SSc) at the onset of respiratory symptoms. Patients and methods: The study population included 19 patients (RA ¼ 12; SSc ¼ 7) and 14 age-and sex-matched healthy volunteers. Lung function testing, high resolution computed tomography (HRCT) of the thorax and IS collection were performed in all cases. Broncho-alveolar lavage (BAL) was obtained in selected patients. Results: IS samples from patients contained a significantly higher percentage of neutrophils and a lower percentage of macrophages compared to healthy subjects (p ¼ 0.002 and 0.001, respectively), while the total cell number showed no differences. In addition, sputa yielded both higher cell counts and higher neutrophils than BAL samples (p ¼ 0.02 in all instances). No correlations were found between IS findings and lung function parameters, HRCT and BAL findings. (A. Vatrella).

Respiratory Medicine, 2010

M. Bocchino). a v a i l a b l e a t w w w . s c i e n c e d i r e c t . c o m j o u r n a l h o m... more M. Bocchino). a v a i l a b l e a t w w w . s c i e n c e d i r e c t . c o m j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / r m e d Respiratory Medicine (2010) 104, 1551e1556 0954-6111/$ -see front matter ª

PLoS ONE, 2009

CD8 T-cells contribute to control of Mycobacterium tuberculosis infection, but little is known ab... more CD8 T-cells contribute to control of Mycobacterium tuberculosis infection, but little is known about the quality of the CD8 Tcell response in subjects with latent infection and in patients with active tuberculosis disease. CD8 T-cells recognizing epitopes from 6 different proteins of Mycobacterium tuberculosis were detected by tetramer staining. Intracellular cytokines staining for specific production of IFN-c and IL-2 was performed, complemented by phenotyping of memory markers on antigen-specific CD8 T-cells. The ex-vivo frequencies of tetramer-specific CD8 T-cells in tuberculous patients before therapy were lower than in subjects with latent infection, but increased at four months after therapy to comparable percentages detected in subjects with latent infection. The majority of CD8 T-cells from subjects with latent infection expressed a terminally-differentiated phenotype (CD45RA + CCR7 2 ). In contrast, tuberculous patients had only 35% of antigen-specific CD8 T-cells expressing this phenotype, while containing higher proportions of cells with an effector memory-and a central memory-like phenotype, and which did not change significantly after therapy. CD8 T-cells from subjects with latent infection showed a codominance of IL-2 + /IFN-c + and IL-2 2 /IFN-c + T-cell populations; interestingly, only the IL-2 + /IFN-c + population was reduced or absent in tuberculous patients, highly suggestive of a restricted functional profile of Mycobacterium tuberculosis-specific CD8 T-cells during active disease. These results suggest distinct Mycobacterium tuberculosis specific CD8 T-cell phenotypic and functional signatures between subjects which control infection (subjects with latent infection) and those who do not (patients with active disease).

Otolaryngology - Head and Neck Surgery, 2003

The Journal of Pathology, 1997

Tuberculosis (TB) is still a major health problem, both as a single disease entity and as a cofac... more Tuberculosis (TB) is still a major health problem, both as a single disease entity and as a cofactor in AIDS. The interaction between macrophage and Mycobacterium tuberculosis (MTB) is a critical step in the establishment of an early chronic infection. This study analyses the capacity of MTB to induce apoptosis in cells obtained by broncho-alveolar lavage (BAL) from patients with reactive pulmonary tuberculosis and from AIDS patients with disseminated pulmonary tuberculosis. Apoptosis was increased three-fold in BAL cells obtained from patients with pulmonary tuberculosis and even more markedly in alveolar macrophages of MTB-infected AIDS patients, compared with controls. Apoptosis was analysed and characterized by propidium iodide (PI) incorporation, terminal deoxy transferase (TDT)-mediated dUTP-biotin nick end labelling (TUNEL), and tissue transglutaminase (tTG) expression. The MTB-macrophage interaction was also investigated in vitro by infecting monocyte-derived macrophages (MDM) with MTB (virulent strain H37Rv). The induction of apoptosis by MTB required viable bacteria, was dose-dependent, and was restricted to H37Rv. Infection with either Mycobacterium avium complex (MAC) or HIV-1 and treatment with heat-killed MTB failed to induce apoptosis.

Infection and Immunity, 2001

Mycobacterium tuberculosis is an intracellular pathogen that readily survives and replicates in h... more Mycobacterium tuberculosis is an intracellular pathogen that readily survives and replicates in human macrophages (M⌽). Host cells have developed different mycobactericidal mechanisms, including the production of inflammatory cytokines. The aim of this study was to compare the M⌽ response, in terms of cytokine gene expression, to infection with the M. tuberculosis laboratory strain H37Rv and the clinical M. tuberculosis isolate CMT97. Both strains induce the production of interleukin-12 (IL-12) and IL-16 at comparable levels. However, the clinical isolate induces a significantly higher and more prolonged M⌽ activation, as shown by reverse transcription-PCR analysis of IL-1␤, IL-6, IL-10, transforming growth factor beta, tumor necrosis factor alpha, and gamma interferon (IFN-␥) transcripts. Interestingly, when IFN-␥ transcription is high, the number of M. tuberculosis genes expressed decreases and vice versa, whereas no mycobactericidal effect was observed in terms of bacterial growth. Expression of 11 genes was also studied in the two M. tuberculosis strains by infecting resting or activated M⌽ and compared to bacterial intracellular survival. In both cases, a peculiar inverse correlation between expression of these genes and multiplication was observed. The number and type of genes expressed by the two strains differed significantly.

Immunology, 2004

This study addresses the analysis of the human leucocyte antigen (HLA) allele distribution in 54 ... more This study addresses the analysis of the human leucocyte antigen (HLA) allele distribution in 54 historical and in 68 recently diagnosed tuberculosis (TB) patients. The historical cohort was characterized by the presence of large ®brocavernous lesions effectively treated with therapeutic pneumothorax during the period 1950±55. Patients and healthy controls enrolled in the study were from the Campania region of southern Italy. No signi®cant association between HLA alleles and TB in the population of recently diagnosed TB patients was observed. On the contrary, among the historical TB patients there was a strong association with an increased frequency of the HLA-DR4 allele alone and/or in the presence of the HLA-B14 allele (P 0Á000004; P c 0Á0008), as well as with a decreased frequency of the HLA-A2 ,-B14 À ,DR4 À allele association (P 0Á00005; P c 0Á01). In order to exclude any interference from age-related factors, these results were con®rmed by comparing the historical cohort of TB patients with an age-matched healthy control population of the same ethnic origin (P 0Á00004; P c 0Á008; and P 0Á0001; and P c 0Á02, respectively).

European Respiratory Journal, 2003

European Journal of Immunology, 2010

Th1 CD4 1 T cells and their derived cytokines are crucial for protection against Mycobacterium tu... more Th1 CD4 1 T cells and their derived cytokines are crucial for protection against Mycobacterium tuberculosis. Using multiparametric flow cytometry, we have evaluated the distribution of seven distinct functional states (IFN-c/IL-2/TNF-a triple expressors, IFN-c/IL-2, IFN-c/TNF-a or TNF-a/IL-2 double expressors or IFN-c, IL-2 or TNF-a single expressors) of CD4 1 T cells in individuals with latent M. tuberculosis infection (LTBI) and active tuberculosis (TB). We found that triple expressors, while detectable in 85-90%TB patients, were only present in 10-15% of LTBI subjects. On the contrary, LTBI subjects had significantly higher (12-to 15-fold) proportions of IL-2/IFN-c double and IFN-c single expressors as compared with the other CD4 1 T-cell subsets. Proportions of the other double or single CD4 1 T-cell expressors did not differ between TB and LTBI subjects. These distinct IFN-c, IL-2 and TNF-a profiles of M. tuberculosis-specific CD4 1 T cells seem to be associated with live bacterial loads, as indicated by the decrease in frequency of multifunctional T cells in TB-infected patients after completion of anti-mycobacterial therapy. Our results suggest that phenotypic and functional signatures of CD4 1 T cells may serve as immunological correlates of protection and curative host responses, and be a useful tool to monitor the efficacy of anti-mycobacterial therapy.

European Journal of Clinical Microbiology & Infectious Diseases, 2008

The reactivation of latent tuberculosis (TB) is a major complication of tumor necrosis factor (TN... more The reactivation of latent tuberculosis (TB) is a major complication of tumor necrosis factor (TNF)-alpha inhibitors. Screening for TB infection is recommended before anti-TNF therapy is initiated; however, the use of tuberculin skin testing (TST) is controversial, due to the high rate of false-negative results in patients receiving immunosuppressive treatment. To compare the performance of two commercial interferon (IFN)-gamma release assays (IGRA), T-SPOT.TB (TS-TB) and QuantiFERON-TB Gold "In-tube" (QFT-GIT), with TST for the detection of TB infection in patients due to start anti-TNF therapy, 69 human immunodeficiency virus (HIV)-negative Italian patients (mean age: 45.2 +/- 12.6 years; male=39) were enrolled between September 2005 to August 2006. Patients affected by rheumatoid arthritis (n = 18), psoriatic arthritis (n = 26), ulcerous rectocolitis (n = 6), and Crohn's disease (n = 19) were tested simultaneously with TST, TS-TB, and QFT-GIT. Overall, 26% of patients were positive by TST, 30.4% by TS-TB, and 31.8% by QFT-GIT. Agreement with TST was similar (kappa = 0.21, p = 0.0002 and kappa = 0.26, p < 0.001, respectively). In 11 TST-negative cases, IFN-gamma release assays were positive. In addition, in seven Mantoux-positive cases with no TB risk factors, TST result agreement was achieved with at least one blood test. Indeterminate results were detected in 5.8% and 2.8% of cases, respectively, with TS-TB and with QFT-GIT (p = not significant [ns]). In conclusion, our results suggest that IGRAs may be helpful for screening purposes in patient candidates for anti-TNF therapy to confirm positive TST results and in selected cases when false-negative results are suspected. The utility of blood tests in patients with low or no TB risk remains to be assessed.

Cytokine, 2014

The immune response plays an unsettled role in the pathogenesis of idiopathic pulmonary fibrosis ... more The immune response plays an unsettled role in the pathogenesis of idiopathic pulmonary fibrosis (IPF), the contribution of inflammation being controversial as well. Emerging novel T cell sub-populations including regulatory T lymphocytes (Treg) and interleukin (IL)-17 secreting T helper cells (Th17) may exert antithetical actions in this scenario. Phenotype and frequency of circulating immune cell subsets were assessed by multi-parametric flow cytometry in 29 clinically stable IPF patients and 17 healthy controls. The interplay between Treg lymphocytes expressing transforming growth factor (TGF)-β and Th17 cells was also investigated. Proportion and absolute number of natural killer (NK) cells were significantly reduced in IPF patients in comparison with controls (p<0.001). Conversely, the proportion and absolute number of CD3(+)CD4(+)CD25(high)Foxp-3(+) cells were significantly increased in IPF patients (p=0.000). As in controls, almost the totality of cells (>90%) expressed TGF-β upon stimulation. Interestingly, the frequency of Th17 cells was significantly compromised in IPF patients (p=0.000) leading to an increased TGF-β/IL-17 ratio (4.2±2.3 vs 0.5±0.3 in controls, p=0.000). Depletion of NK and Th17 cells along with a not compromised Treg compartment delineate the existence of an "immune profile" that argue against the recent hypothesis of IPF as an autoimmune disease. Our findings along with the imbalance of the Treg/Th17 axis more closely suggest these immune perturbations to be similar to those observed in cancer. Clinical relevance, limitations and perspectives for future research are discussed.

Clinical Rheumatology, 2014

Clinical Rheumatology, 2010

The aim of this study