Human proteomic profiles in latent and active tuberculosis

Distinguishing patients with active tuberculosis (TB) from those with latent TB is an important clinical problem. The SELDI-TOF MS (Surface Enhanced Laser Desorption Ionisation – Time of Flight Mass Spectrometry) platform allows for high throughput detection of multiple proteins in biological fluids. Proteomic patterns reflecting host-pathogen interaction can be used as a tool to aid our understanding of the Natural History of Tuberculosis. Methods: Plasma samples were collected prospectively in a shanty town in Lima, Peru. Latent and active TB status was defined using the Tuberculin Skin Test (TST), Quantiferon (QFN) assay and TB culture. Crude plasma and fractionated plasma samples were analysed on weak cationic CM10 chip surfaces using a Biomek 3000 Laboratory Automation Workstation. Spectra were generated using a ProteinChip System 4000 Mass spectrometer. Data was analysed using a Support Vector Machine. Results: Samples were collected from 154 patients with active TB, 112 patients with respiratory symptoms suggestive of TB and 151 healthy controls. Multiple peaks differed significantly between active TB patients and unhealthy controls. Trained optimal classifiers discriminate between: i) active TB and unhealthy controls with 84% accuracy (87% sensitivity, 79% specificity) in crude plasma and up to 89% accuracy (90% sensitivity, 88% specificity) in fractionated plasma ii) active TB and latent TB with 89% accuracy (90% sensitivity, 89% specificity) iii) latent TB and no TB in healthy controls with 77% accuracy (67% sensitivity, 84% specificity). Conclusions: SELDI-TOF MS proteomic profiles in combination with trained optimal classifiers accurately discriminate active TB from other respiratory disorders. The classifier for latent TB was not as accurate, but active TB could be discriminated from latent TB

Gender-dependent differences in plasma matrix metalloproteinase-8 elevated in pulmonary tuberculosis.

Tuberculosis (TB) remains a global health pandemic and greater understanding of underlying pathogenesis is required to develop novel therapeutic and diagnostic approaches. Matrix metalloproteinases (MMPs) are emerging as key effectors of tissue destruction in TB but have not been comprehensively studied in plasma, nor have gender differences been investigated. We measured the plasma concentrations of MMPs in a carefully characterised, prospectively recruited clinical cohort of 380 individuals. The collagenases, MMP-1 and MMP-8, were elevated in plasma of patients with pulmonary TB relative to healthy controls, and MMP-7 (matrilysin) and MMP-9 (gelatinase B) were also increased. MMP-8 was TB-specific (p<0.001), not being elevated in symptomatic controls (symptoms suspicious of TB but active disease excluded). Plasma MMP-8 concentrations inversely correlated with body mass index. Plasma MMP-8 concentration was 1.51-fold higher in males than females with TB (p<0.05) and this difference was not due to greater disease severity in men. Gender-specific analysis of MMPs demonstrated consistent increase in MMP-1 and -8 in TB, but MMP-8 was a better discriminator for TB in men. Plasma collagenases are elevated in pulmonary TB and differ between men and women. Gender must be considered in investigation of TB immunopathology and development of novel diagnostic markers

Impact on and use of an inner-city London Infectious Diseases Department by international migrants: a questionnaire survey

<p>Abstract</p> <p>Background</p> <p>The UK has witnessed a considerable increase in immigration in the past decade. Migrant may face barriers to accessing appropriate health care on arrival and the current focus on screening certain migrants for tuberculosis on arrival is considered inadequate. We assessed the implications for an inner-city London Infectious Diseases Department in a high migrant area.</p> <p>Methods</p> <p>We administered an anonymous 20-point questionnaire survey to all admitted patients during a 6 week period. Questions related to sociodemographic characteristics and clinical presentation. Analysis was by migration status (UK born <it>vs </it>overseas born).</p> <p>Results</p> <p>111 of 133 patients completed the survey (response rate 83.4%). 58 (52.2%) were born in the UK; 53 (47.7%) of the cohort were overseas born. Overseas-born were over-represented in comparison to Census data for this survey site (47.7% <it>vs </it>33.6%; proportional difference 0.142 [95% CI 0.049–0.235]; p = 0.002): overseas born reported 33 different countries of birth, most (73.6%) of whom arrived in the UK pre-1975 and self-reported their nationality as British. A smaller number (26.4%) were new migrants to the UK (≤10 years), mostly refugees/asylum seekers. Overseas-born patients presented with a broad range and more severe spectrum of infections, differing from the UK-born population, resulting in two deaths in this group only. Presentation with a primary infection was associated with refugee/asylum status (n = 8; OR 6.35 [95% CI 1.28–31.50]; p = 0.023), being a new migrant (12; 10.62 [2.24–50.23]; p = 0.003), and being overseas born (31; 3.69 [1.67–8.18]; p = 0.001). Not having registered with a primary-care physician was associated with being overseas born, being a refugee/asylum seeker, being a new migrant, not having English as a first language, and being in the UK for ≤5 years. No significant differences were found between groups in terms of duration of illness prior to presentation or duration of hospitalisation (mean 11.74 days [SD 12.69]).</p> <p>Conclusion</p> <p>Migrants presented with a range of more severe infections, which suggests they face barriers to accessing appropriate health care and screening both on arrival and once settled through primary care services. A more organised and holistic approach to migrant health care is required.</p

Discriminating Active from Latent Tuberculosis in Patients Presenting to Community Clinics

BACKGROUND Because of the high global prevalence of latent TB infection (LTBI), a key challenge in endemic settings is distinguishing patients with active TB from patients with overlapping clinical symptoms without active TB but with co-existing LTBI. Current methods are insufficiently accurate. Plasma proteomic fingerprinting can resolve this difficulty by providing a molecular snapshot defining disease state that can be used to develop point-of-care diagnostics. METHODS Plasma and clinical data were obtained prospectively from patients attending community TB clinics in Peru and from household contacts. Plasma was subjected to high-throughput proteomic profiling by mass spectrometry. Statistical pattern recognition methods were used to define mass spectral patterns that distinguished patients with active TB from symptomatic controls with or without LTBI. RESULTS 156 patients with active TB and 110 symptomatic controls (patients with respiratory symptoms without active TB) were investigated. Active TB patients were distinguishable from undifferentiated symptomatic controls with accuracy of 87% (sensitivity 84%, specificity 90%), from symptomatic controls with LTBI (accuracy of 87%, sensitivity 89%, specificity 82%) and from symptomatic controls without LTBI (accuracy 90%, sensitivity 90%, specificity 92%). CONCLUSIONS We show that active TB can be distinguished accurately from LTBI in symptomatic clinic attenders using a plasma proteomic fingerprint. Translation of biomarkers derived from this study into a robust and affordable point-of-care format will have significant implications for recognition and control of active TB in high prevalence settings

Human proteomic profiles in latent and active tuberculosis

Distinguishing patients with active tuberculosis (TB) from those with latent TB is an important clinical problem. The SELDI-TOF MS (Surface Enhanced Laser Desorption Ionisation – Time of Flight Mass Spectrometry) platform allows for high throughput detection of multiple proteins in biological fluids. Proteomic patterns reflecting host-pathogen interaction can be used as a tool to aid our understanding of the Natural History of Tuberculosis. Methods: Plasma samples were collected prospectively in a shanty town in Lima, Peru. Latent and active TB status was defined using the Tuberculin Skin Test (TST), Quantiferon (QFN) assay and TB culture. Crude plasma and fractionated plasma samples were analysed on weak cationic CM10 chip surfaces using a Biomek 3000 Laboratory Automation Workstation. Spectra were generated using a ProteinChip System 4000 Mass spectrometer. Data was analysed using a Support Vector Machine. Results: Samples were collected from 154 patients with active TB, 112 patients with respiratory symptoms suggestive of TB and 151 healthy controls. Multiple peaks differed significantly between active TB patients and unhealthy controls. Trained optimal classifiers discriminate between: i) active TB and unhealthy controls with 84% accuracy (87% sensitivity, 79% specificity) in crude plasma and up to 89% accuracy (90% sensitivity, 88% specificity) in fractionated plasma ii) active TB and latent TB with 89% accuracy (90% sensitivity, 89% specificity) iii) latent TB and no TB in healthy controls with 77% accuracy (67% sensitivity, 84% specificity). Conclusions: SELDI-TOF MS proteomic profiles in combination with trained optimal classifiers accurately discriminate active TB from other respiratory disorders. The classifier for latent TB was not as accurate, but active TB could be discriminated from latent TB.EThOS - Electronic Theses Online ServiceWellcome TrustGBUnited Kingdo

Severe bilateral knee osteonecrosis in a young man with human immunodeficiency virus

We present a young man with a background Human Immunodeficiency virus (HIV) who presented with bilateral knee pain and reduced mobility. Subsequent imaging of the knees demonstrated florid osteonecrosis (ON), which was managed conservatively. ON is seen more commonly in HIV patients than the general population, however the underlying mechanism for this association is remains unclear. An awareness of this disease is imperative to appropriately identify and manage such patients. Keywords: Osteonecrosis, Musculoskeletal disorders, HIV / AID

Neutrophil collagenase MMP-8 is specifically upregulated in the plasma in active pulmonary tuberculosis (TB).

<p>Plasma samples were collected prospectively from patients in Lima, Peru. MMP concentrations were measured by Luminex bead array. <b>A</b>. MMP-1 was increased in both active TB and respiratory symptomatics compared to healthy controls (p<0.001 and 0.01 respectively). <b>B</b>. MMP-3 showed no difference between the groups. <b>C</b>. MMP-7 was increased in both active TB and symptomatic controls compared to healthy controls (p<0.01 for both). <b>D</b>. MMP-8 was specifically increased in active TB compared to both respiratory symptomatics and healthy controls (p<0.001 for both). <b>E</b>. MMP-9 was increased in active TB only compared to healthy controls (p<0.05). <b>F</b>. MMP-10 showed no difference between the groups. Statistical analysis was performed using a one way ANOVA with Tukey’s post hoc test, ***p<0.001, **p<0.01, *p<0.05. Each box represents the 25^th to 75^th centiles, the central line the median, and the whiskers the minimum and maximum values.</p