Dino Tan | The University of Western Australia (original) (raw)

Papers by Dino Tan

Research paper thumbnail of A simplified protocol for profiling heparin-contaminated circulating miRNAs: by microfluidic array

Molecular Biology Reports, 2020

Peripheral blood is a valuable, non-invasive source of biomarkers which include circulating miRNA... more Peripheral blood is a valuable, non-invasive source of biomarkers which include circulating miRNAs. Using microfluidic array-based techniques, miRNAs can be successfully measured in small amounts of blood plasma (< 0.5 mL) using cDNA pre-amplification. However, the use of heparin-based anticoagulants for blood collection hinders the detection of circulating miRNAs due to its inhibitory effect on PCR components. Although pre-treatment with heparinase have been shown to overcome heparin contamination in blood, its effect has not been described in array-based analyses or more sensitive applications with smaller sample volumes (i.e. 200 μL plasma) requiring pre-amplification. We show that the treatment of miRNA extracted from heparinised plasma with an optimised concentration of Bacteroides heparinase I prior to cDNA preamplification dramatically improves the number of detectable miRNA from 2 to 67 targets on the TaqMan ® Array Human MicroRNA Cards. Furthermore, the titrated amount of heparinase (3 U) gave the best miRNA detection compared to those used in previous studies (6-24 U). This study provides novel data which demonstrates that heparinase treatment is compatible with protocols that involve pre-amplification of cDNA and microfluidic array-based techniques. This an improved methodology that permits miRNA-based biomarker analysis from small volume of heparinised plasma.

Research paper thumbnail of Immune Dysregulation in HIV patients following successful ART

Research paper thumbnail of The role of CMV infection in COPD Levels of CMV-reactive antibodies correlate with the induction of CD 28 null T-cells and systemic inflammation in chronic obstructive pulmonary disease ( COPD )

Research paper thumbnail of Impaired CTLA-4 responses in COPD are associated with systemic inflammation

Cellular & molecular immunology, 2014

Research paper thumbnail of Impaired function of regulatory T-cells in patients with chronic obstructive pulmonary disease (COPD)

Immunobiology, 2014

Anti-inflammatory pathways affecting chronic obstructive pulmonary disease (COPD) are poorly unde... more Anti-inflammatory pathways affecting chronic obstructive pulmonary disease (COPD) are poorly understood. Regulatory T-cells (Tregs) are important negative regulators of T-cell activity and hence were investigated in COPD patients in this study. We hypothesised that functional defects in Tregs may promote increased inflammation contributing to the pathogenesis of COPD. Peripheral blood mononuclear cells (PBMC) were isolated from patients with stable COPD and age-matched non-smoking controls. Treg-mediated suppression of memory non-Treg (Foxp3(-)CD45RO(+)) CD4(+) T-cell activation was analysed by comparing PBMC responses to staphylococcal enterotoxin-B (SEB) pre- and post-depletion of Tregs (CD25(+)CD127(low)CD4(+) T-cells) by fluorescence-activated cell sorting (FACS). Activation of T-cells was assessed by HLA-DR expression. Levels of secreted cytokines were measured by ELISA. Depletion of Tregs increased SEB-induced activation of Foxp3(-)CD45RO(+) CD4(+) T-cells in samples from 15/1...

Research paper thumbnail of Levels of CMV-reactive antibodies correlate with the induction of CD28(null) T cells and systemic inflammation in chronic obstructive pulmonary disease (COPD)

Cellular & molecular immunology, Jan 26, 2015

Research paper thumbnail of Longitudinal Plasma Antibody Titers in Relation to IRD in HIV Patients Beginning ART

International Journal of Infectious Diseases, 2008

e202 13th International Congress on Infectious Diseases Abstracts, Poster Presentations 222) befo... more e202 13th International Congress on Infectious Diseases Abstracts, Poster Presentations 222) before, and 153 cells/microliter (IQR, 49-187; p = 0.3) after DAART. Among those who no longer met the treatment failure criteria after DAART, the median CD4 counts were 150 cells/microliter (IQR, 113-233) before, and 244 cells/microliter (IQR, 132-287; p = 0.02) after DAART. At the time of this analysis, the women who responded to DAART have remained on first-line therapy without additional episodes of treatment failure for a median of 170 (IQR, 57-450) days. Conclusions: One month of DAART with adherence counseling could provide a simple, low-cost method of optimizing the durability of response to first-line ART and distinguishing faltering adherence from treatment failure.

Research paper thumbnail of The proportion and function of peripheral myeloid-derived suppressor cells do not correlate with systemic inflammation in chronic obstructive pulmonary disease

Human Immunology, 2014

Myeloid-derived suppressor cells (MDSC) have been implicated in the regulation of chronic inflamm... more Myeloid-derived suppressor cells (MDSC) have been implicated in the regulation of chronic inflammation. Chronic obstructive pulmonary disease (COPD) involves persistent inflammation, but the role of MDSC has not been explored. Here, proportions of MDSC (CD14 À HLA-DR À CD33 + CD11b + cells) were quantified in peripheral blood mononuclear cells (PBMC) isolated from patients with 'stable' COPD (n = 12), smokers with no evidence of COPD (n = 11) and healthy non-smokers (n = 11). The proportions of MDSC were similar in all groups. MDSC function was assessed by comparing T-cell and cytokine responses of whole and MDSC-depleted PBMC stimulated with Staphylococcus enterotoxin-B (SEB). Depletion of MDSC did not enhance CD4 + or CD8 + T-cell activation and proliferation, or alter IFNc and IL-17 production in response to SEB. However production of TGFb decreased after depletion of MDSC, so MDSC may be a source of this cytokine. In conclusion, COPD was not associated with perturbations in the proportion or function of MDSC in peripheral blood.

Research paper thumbnail of Levels of anti-cytokine antibodies may be elevated in patients with pulmonary disease associated with non-tuberculous mycobacteria

Cytokine, 2014

Pulmonary disease due to non-tuberculous mycobacteria (NTM) is caused by several species (particu... more Pulmonary disease due to non-tuberculous mycobacteria (NTM) is caused by several species (particularly Mycobacterium avium, Mycobacterium intracellulare) that are abundant in the environment. Th1 cytokines such as interferon (IFN)-γ are important in the control of mycobacteria, but in vitro production of IFN-γ is not deficient in adult patients with pulmonary NTM disease. Antibodies reactive with IFN-γ have been described in patients with disseminated NTM disease, but it is not clear whether they are common in pulmonary disease. Here we show that patients with pulmonary NTM have a higher level of anti-IFN-γ and anti-GM-CSF antibodies than healthy controls, although some controls also have high levels. Levels of anti-IFN-γ antibodies did not correlate with levels of total immunoglobulin. Longitudinal studies are required to determine whether anti-cytokine autoantibodies are consequence rather than a cause of pulmonary NTM disease.

Research paper thumbnail of Robust interferon-α and IL-12 responses by dendritic cells are related to efficient CD4+ T-cell recovery in HIV patients on ART

Clinical Immunology, 2011

Amongst HIV patients with successful virological responses to antiretroviral therapy (ART), poor ... more Amongst HIV patients with successful virological responses to antiretroviral therapy (ART), poor CD4 + T-cell recovery is associated with low nadir CD4 + T-cell counts and persistent immune activation. These factors might be influenced by dendritic cell (DC) function. Interferonα-producing plasmacytoid DC and IL-12-producing myeloid DC were quantified by flow cytometry after stimulation with agonists to TLR7/8 (CL075) or TLR9 (CpG-ODN). These were compared between patients who achieved CD4 + T-cell counts above or below 200 cells/μL after 6 months on ART (High vs. Low groups). High Group patients had more DC producing interferon-α or IL-12 at Weeks 6 and 12 on ART than Low Group patients. The frequencies of cytokine-producing DC at Week 12 were directly correlated with CD4 + T-cell counts at baseline and at Week 12. Patients with good recovery of CD4 + T-cells had robust TLR-mediated interferon-α responses by plasmacytoid DC and IL-12 responses by myeloid DC during early ART (1-3 months).

Research paper thumbnail of Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high on antiretroviral therapy

AIDS, 2007

Objective: To examine the relationships between blood CD4 natural regulatory T (T reg) cells, pla... more Objective: To examine the relationships between blood CD4 natural regulatory T (T reg) cells, plasma HIV RNA level, CD4 T-cell count and immune activation in untreated HIV-infected patients and immunodeficient patients beginning antiretroviral therapy (ART), using a novel phenotype to define T reg cells (CD25CD127 lo CD4). Data were compared with established T reg cell markers (FoxP3, CTLA-4 and GITR). Methods: Twenty-nine untreated HIV-infected patients with CD4 T-cell counts of < 300 or > 400/ml were compared in a cross-sectional study and 12 patients beginning combination ART with < 100 CD4 T cells/ml were followed for 1 year on therapy. Three-and four-colour flow cytometry was used to quantitate proportions of T reg cells. Results: In control donors and patients with high CD4 T-cell counts, 28-89% (median 60%) of CD25CD127 lo CD4 cells were FoxP3, but < 10% expressed GITR or CTLA-4. Immunodeficient patients also had CD4-negative lymphocytes with the phenotype FoxP3CD127 lo. Proportions of CD25CD127 lo cells and activated (HLA-DR hi) cells in the CD4 T-cell population were increased in patients with low CD4 T cell counts. The proportion of CD25CD127 lo CD4 T cells correlated positively with plasma HIV RNA level and CD4 T-cell activation, but inversely with CD4 T-cell count. Longitudinal studies of 12 patients receiving ART in two distinct cohorts (Western Australia and Malaysia) showed that the proportion of CD25CD127 lo CD4 cells decreased slightly over time, but remained above levels seen in non-HIV controls. Conclusions: Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high after 1 year on ART.

Research paper thumbnail of There is detectable variation in the lipidomic profile between stable and progressive patients with idiopathic pulmonary fibrosis (IPF)

Respiratory Research, 2021

Background Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease characteriz... more Background Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease characterized by fibrosis and progressive loss of lung function. The pathophysiological pathways involved in IPF are not well understood. Abnormal lipid metabolism has been described in various other chronic lung diseases including asthma and chronic obstructive pulmonary disease (COPD). However, its potential role in IPF pathogenesis remains unclear. Methods In this study, we used ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) to characterize lipid changes in plasma derived from IPF patients with stable and progressive disease. We further applied a data-independent acquisition (DIA) technique called SONAR, to improve the specificity of lipid identification. Results Statistical modelling showed variable discrimination between the stable and progressive subjects, revealing differences in the detection of triglycerides (TG) and phosphatidylcholines ...

Research paper thumbnail of Transcriptional profiling of circulating mononuclear cells from patients with chronic obstructive pulmonary disease receiving mesenchymal stromal cell infusions

STEM CELLS Translational Medicine

Research paper thumbnail of Untargeted metabolomics of human plasma reveal lipid markers unique to chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis

PROTEOMICS – Clinical Applications

Research paper thumbnail of Circulating RNA differences between patients with stable and progressive idiopathic pulmonary fibrosis

European Respiratory Journal

Research paper thumbnail of Early cessation of a randomised study in acute graft versus host disease: upfront mesenchymal stromal cells with corticosteroids versus corticosteroids alone

Bone Marrow Transplantation

Research paper thumbnail of Impaired Th1 responses in patients with acute exacerbations of COPD are improved with PD-1 blockade

Research paper thumbnail of Evidence of functional cell-mediated immune responses to nontypeable Haemophilus influenzae in otitis-prone children

PLOS ONE

Otitis media (OM) remains a common paediatric disease, despite advances in vaccinology. Susceptib... more Otitis media (OM) remains a common paediatric disease, despite advances in vaccinology. Susceptibility to recurrent acute OM (rAOM) has been postulated to involve defective cell-mediated immune responses to common otopathogenic bacteria. We compared the composition of peripheral blood mononuclear cells (PBMC) from 20 children with a history of rAOM (otitis-prone) and 20 healthy non-otitis-prone controls, and assessed innate and cell-mediated immune responses to the major otopathogen nontypeable Haemophilus influenzae (NTHi). NTHi was a potent stimulator of inflammatory cytokine secretion from PBMC within 4 hours, with no difference in cytokine levels produced between PBMC from cases or controls. In the absence of antigen stimulation, otitis-prone children had more circulating Natural Killer (NK) cells (p<0.01), particularly NK dim (CD56 lo) cells (p<0.01), but fewer CD4 + T cells (p<0.01) than healthy controls. NTHi challenge significantly increased the proportion of activated (CD107a +) NK cells in otitis-prone and non-otitis-prone children (p<0.01), suggesting that NK cells from otitis-prone children are functional and respond to NTHi. CD8 + T cells and NK cells from both cases and controls produced IFNγ in response to polyclonal stimulus (Staphylococcal enterotoxin B; SEB), with more IFNγ + CD8 + T cells present in cases than controls (p<0.05) but similar proportions of IFNγ + NK cells. Otitisprone children had more circulating IFNγ-producing NK cells (p<0.05) and more IFNγ-producing CD4 + (p<0.01) or CD8 + T-cells (p<0.05) than healthy controls. In response to SEB, more CD107a-expressing CD8 + T cells were present in cases than controls (p<0.01). Despite differences in PBMC composition, PBMC from otitis-prone children mounted innate and T cell-mediated responses to NTHi challenge that were comparable to healthy children. These data provide evidence that otitis-prone children do not have impaired functional cell mediated immunity.

Research paper thumbnail of Analysis by proteomics reveals unique circulatory proteins in idiopathic pulmonary fibrosis

Respirology

Background and objective: Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic disease t... more Background and objective: Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic disease that has a poor 3-year median survival rate with unclear pathophysiology. Radiological features include bibasal, subpleural fibrosis and honeycombing while its pathology is characterized by fibroblastic foci and honeycombing. Proteomic analysis of circulating molecules in plasma may identify factors that characterize IPF and may assist in the diagnosis, prognostication and determination of pathogenic pathways in this condition. Methods: Two independent quantitative proteomic techniques were used, isobaric tags for relative and absolute quantitation (iTRAQ) and multiple reaction monitoring (MRM), to identify differentially expressed plasma proteins in a group of IPF patients in comparison to healthy controls with normal lung function matched for age and gender. Results: Five proteins were identified to be differentially expressed in IPF compared to healthy controls (upregulation of platelet basic protein and downregulation of actin, cytoplasmic 2, antithrombin-III, extracellular matrix protein-1 and fibronectin). Conclusion: This study further validates the combinational use of non-targeted discovery proteomics (iTRAQ) with targeted quantitation by mass spectrometry (MRM) of soluble biomarkers to identify potentially important molecules and pathways for pulmonary diseases such as IPF.

Research paper thumbnail of Circulating RNA differences between patients with stable and progressive lung function in IPF

European Respiratory Journal

Research paper thumbnail of A simplified protocol for profiling heparin-contaminated circulating miRNAs: by microfluidic array

Molecular Biology Reports, 2020

Peripheral blood is a valuable, non-invasive source of biomarkers which include circulating miRNA... more Peripheral blood is a valuable, non-invasive source of biomarkers which include circulating miRNAs. Using microfluidic array-based techniques, miRNAs can be successfully measured in small amounts of blood plasma (< 0.5 mL) using cDNA pre-amplification. However, the use of heparin-based anticoagulants for blood collection hinders the detection of circulating miRNAs due to its inhibitory effect on PCR components. Although pre-treatment with heparinase have been shown to overcome heparin contamination in blood, its effect has not been described in array-based analyses or more sensitive applications with smaller sample volumes (i.e. 200 μL plasma) requiring pre-amplification. We show that the treatment of miRNA extracted from heparinised plasma with an optimised concentration of Bacteroides heparinase I prior to cDNA preamplification dramatically improves the number of detectable miRNA from 2 to 67 targets on the TaqMan ® Array Human MicroRNA Cards. Furthermore, the titrated amount of heparinase (3 U) gave the best miRNA detection compared to those used in previous studies (6-24 U). This study provides novel data which demonstrates that heparinase treatment is compatible with protocols that involve pre-amplification of cDNA and microfluidic array-based techniques. This an improved methodology that permits miRNA-based biomarker analysis from small volume of heparinised plasma.

Research paper thumbnail of Immune Dysregulation in HIV patients following successful ART

Research paper thumbnail of The role of CMV infection in COPD Levels of CMV-reactive antibodies correlate with the induction of CD 28 null T-cells and systemic inflammation in chronic obstructive pulmonary disease ( COPD )

Research paper thumbnail of Impaired CTLA-4 responses in COPD are associated with systemic inflammation

Cellular & molecular immunology, 2014

Research paper thumbnail of Impaired function of regulatory T-cells in patients with chronic obstructive pulmonary disease (COPD)

Immunobiology, 2014

Anti-inflammatory pathways affecting chronic obstructive pulmonary disease (COPD) are poorly unde... more Anti-inflammatory pathways affecting chronic obstructive pulmonary disease (COPD) are poorly understood. Regulatory T-cells (Tregs) are important negative regulators of T-cell activity and hence were investigated in COPD patients in this study. We hypothesised that functional defects in Tregs may promote increased inflammation contributing to the pathogenesis of COPD. Peripheral blood mononuclear cells (PBMC) were isolated from patients with stable COPD and age-matched non-smoking controls. Treg-mediated suppression of memory non-Treg (Foxp3(-)CD45RO(+)) CD4(+) T-cell activation was analysed by comparing PBMC responses to staphylococcal enterotoxin-B (SEB) pre- and post-depletion of Tregs (CD25(+)CD127(low)CD4(+) T-cells) by fluorescence-activated cell sorting (FACS). Activation of T-cells was assessed by HLA-DR expression. Levels of secreted cytokines were measured by ELISA. Depletion of Tregs increased SEB-induced activation of Foxp3(-)CD45RO(+) CD4(+) T-cells in samples from 15/1...

Research paper thumbnail of Levels of CMV-reactive antibodies correlate with the induction of CD28(null) T cells and systemic inflammation in chronic obstructive pulmonary disease (COPD)

Cellular & molecular immunology, Jan 26, 2015

Research paper thumbnail of Longitudinal Plasma Antibody Titers in Relation to IRD in HIV Patients Beginning ART

International Journal of Infectious Diseases, 2008

e202 13th International Congress on Infectious Diseases Abstracts, Poster Presentations 222) befo... more e202 13th International Congress on Infectious Diseases Abstracts, Poster Presentations 222) before, and 153 cells/microliter (IQR, 49-187; p = 0.3) after DAART. Among those who no longer met the treatment failure criteria after DAART, the median CD4 counts were 150 cells/microliter (IQR, 113-233) before, and 244 cells/microliter (IQR, 132-287; p = 0.02) after DAART. At the time of this analysis, the women who responded to DAART have remained on first-line therapy without additional episodes of treatment failure for a median of 170 (IQR, 57-450) days. Conclusions: One month of DAART with adherence counseling could provide a simple, low-cost method of optimizing the durability of response to first-line ART and distinguishing faltering adherence from treatment failure.

Research paper thumbnail of The proportion and function of peripheral myeloid-derived suppressor cells do not correlate with systemic inflammation in chronic obstructive pulmonary disease

Human Immunology, 2014

Myeloid-derived suppressor cells (MDSC) have been implicated in the regulation of chronic inflamm... more Myeloid-derived suppressor cells (MDSC) have been implicated in the regulation of chronic inflammation. Chronic obstructive pulmonary disease (COPD) involves persistent inflammation, but the role of MDSC has not been explored. Here, proportions of MDSC (CD14 À HLA-DR À CD33 + CD11b + cells) were quantified in peripheral blood mononuclear cells (PBMC) isolated from patients with 'stable' COPD (n = 12), smokers with no evidence of COPD (n = 11) and healthy non-smokers (n = 11). The proportions of MDSC were similar in all groups. MDSC function was assessed by comparing T-cell and cytokine responses of whole and MDSC-depleted PBMC stimulated with Staphylococcus enterotoxin-B (SEB). Depletion of MDSC did not enhance CD4 + or CD8 + T-cell activation and proliferation, or alter IFNc and IL-17 production in response to SEB. However production of TGFb decreased after depletion of MDSC, so MDSC may be a source of this cytokine. In conclusion, COPD was not associated with perturbations in the proportion or function of MDSC in peripheral blood.

Research paper thumbnail of Levels of anti-cytokine antibodies may be elevated in patients with pulmonary disease associated with non-tuberculous mycobacteria

Cytokine, 2014

Pulmonary disease due to non-tuberculous mycobacteria (NTM) is caused by several species (particu... more Pulmonary disease due to non-tuberculous mycobacteria (NTM) is caused by several species (particularly Mycobacterium avium, Mycobacterium intracellulare) that are abundant in the environment. Th1 cytokines such as interferon (IFN)-γ are important in the control of mycobacteria, but in vitro production of IFN-γ is not deficient in adult patients with pulmonary NTM disease. Antibodies reactive with IFN-γ have been described in patients with disseminated NTM disease, but it is not clear whether they are common in pulmonary disease. Here we show that patients with pulmonary NTM have a higher level of anti-IFN-γ and anti-GM-CSF antibodies than healthy controls, although some controls also have high levels. Levels of anti-IFN-γ antibodies did not correlate with levels of total immunoglobulin. Longitudinal studies are required to determine whether anti-cytokine autoantibodies are consequence rather than a cause of pulmonary NTM disease.

Research paper thumbnail of Robust interferon-α and IL-12 responses by dendritic cells are related to efficient CD4+ T-cell recovery in HIV patients on ART

Clinical Immunology, 2011

Amongst HIV patients with successful virological responses to antiretroviral therapy (ART), poor ... more Amongst HIV patients with successful virological responses to antiretroviral therapy (ART), poor CD4 + T-cell recovery is associated with low nadir CD4 + T-cell counts and persistent immune activation. These factors might be influenced by dendritic cell (DC) function. Interferonα-producing plasmacytoid DC and IL-12-producing myeloid DC were quantified by flow cytometry after stimulation with agonists to TLR7/8 (CL075) or TLR9 (CpG-ODN). These were compared between patients who achieved CD4 + T-cell counts above or below 200 cells/μL after 6 months on ART (High vs. Low groups). High Group patients had more DC producing interferon-α or IL-12 at Weeks 6 and 12 on ART than Low Group patients. The frequencies of cytokine-producing DC at Week 12 were directly correlated with CD4 + T-cell counts at baseline and at Week 12. Patients with good recovery of CD4 + T-cells had robust TLR-mediated interferon-α responses by plasmacytoid DC and IL-12 responses by myeloid DC during early ART (1-3 months).

Research paper thumbnail of Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high on antiretroviral therapy

AIDS, 2007

Objective: To examine the relationships between blood CD4 natural regulatory T (T reg) cells, pla... more Objective: To examine the relationships between blood CD4 natural regulatory T (T reg) cells, plasma HIV RNA level, CD4 T-cell count and immune activation in untreated HIV-infected patients and immunodeficient patients beginning antiretroviral therapy (ART), using a novel phenotype to define T reg cells (CD25CD127 lo CD4). Data were compared with established T reg cell markers (FoxP3, CTLA-4 and GITR). Methods: Twenty-nine untreated HIV-infected patients with CD4 T-cell counts of < 300 or > 400/ml were compared in a cross-sectional study and 12 patients beginning combination ART with < 100 CD4 T cells/ml were followed for 1 year on therapy. Three-and four-colour flow cytometry was used to quantitate proportions of T reg cells. Results: In control donors and patients with high CD4 T-cell counts, 28-89% (median 60%) of CD25CD127 lo CD4 cells were FoxP3, but < 10% expressed GITR or CTLA-4. Immunodeficient patients also had CD4-negative lymphocytes with the phenotype FoxP3CD127 lo. Proportions of CD25CD127 lo cells and activated (HLA-DR hi) cells in the CD4 T-cell population were increased in patients with low CD4 T cell counts. The proportion of CD25CD127 lo CD4 T cells correlated positively with plasma HIV RNA level and CD4 T-cell activation, but inversely with CD4 T-cell count. Longitudinal studies of 12 patients receiving ART in two distinct cohorts (Western Australia and Malaysia) showed that the proportion of CD25CD127 lo CD4 cells decreased slightly over time, but remained above levels seen in non-HIV controls. Conclusions: Proportions of circulating T cells with a regulatory cell phenotype increase with HIV-associated immune activation and remain high after 1 year on ART.

Research paper thumbnail of There is detectable variation in the lipidomic profile between stable and progressive patients with idiopathic pulmonary fibrosis (IPF)

Respiratory Research, 2021

Background Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease characteriz... more Background Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease characterized by fibrosis and progressive loss of lung function. The pathophysiological pathways involved in IPF are not well understood. Abnormal lipid metabolism has been described in various other chronic lung diseases including asthma and chronic obstructive pulmonary disease (COPD). However, its potential role in IPF pathogenesis remains unclear. Methods In this study, we used ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) to characterize lipid changes in plasma derived from IPF patients with stable and progressive disease. We further applied a data-independent acquisition (DIA) technique called SONAR, to improve the specificity of lipid identification. Results Statistical modelling showed variable discrimination between the stable and progressive subjects, revealing differences in the detection of triglycerides (TG) and phosphatidylcholines ...

Research paper thumbnail of Transcriptional profiling of circulating mononuclear cells from patients with chronic obstructive pulmonary disease receiving mesenchymal stromal cell infusions

STEM CELLS Translational Medicine

Research paper thumbnail of Untargeted metabolomics of human plasma reveal lipid markers unique to chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis

PROTEOMICS – Clinical Applications

Research paper thumbnail of Circulating RNA differences between patients with stable and progressive idiopathic pulmonary fibrosis

European Respiratory Journal

Research paper thumbnail of Early cessation of a randomised study in acute graft versus host disease: upfront mesenchymal stromal cells with corticosteroids versus corticosteroids alone

Bone Marrow Transplantation

Research paper thumbnail of Impaired Th1 responses in patients with acute exacerbations of COPD are improved with PD-1 blockade

Research paper thumbnail of Evidence of functional cell-mediated immune responses to nontypeable Haemophilus influenzae in otitis-prone children

PLOS ONE

Otitis media (OM) remains a common paediatric disease, despite advances in vaccinology. Susceptib... more Otitis media (OM) remains a common paediatric disease, despite advances in vaccinology. Susceptibility to recurrent acute OM (rAOM) has been postulated to involve defective cell-mediated immune responses to common otopathogenic bacteria. We compared the composition of peripheral blood mononuclear cells (PBMC) from 20 children with a history of rAOM (otitis-prone) and 20 healthy non-otitis-prone controls, and assessed innate and cell-mediated immune responses to the major otopathogen nontypeable Haemophilus influenzae (NTHi). NTHi was a potent stimulator of inflammatory cytokine secretion from PBMC within 4 hours, with no difference in cytokine levels produced between PBMC from cases or controls. In the absence of antigen stimulation, otitis-prone children had more circulating Natural Killer (NK) cells (p<0.01), particularly NK dim (CD56 lo) cells (p<0.01), but fewer CD4 + T cells (p<0.01) than healthy controls. NTHi challenge significantly increased the proportion of activated (CD107a +) NK cells in otitis-prone and non-otitis-prone children (p<0.01), suggesting that NK cells from otitis-prone children are functional and respond to NTHi. CD8 + T cells and NK cells from both cases and controls produced IFNγ in response to polyclonal stimulus (Staphylococcal enterotoxin B; SEB), with more IFNγ + CD8 + T cells present in cases than controls (p<0.05) but similar proportions of IFNγ + NK cells. Otitisprone children had more circulating IFNγ-producing NK cells (p<0.05) and more IFNγ-producing CD4 + (p<0.01) or CD8 + T-cells (p<0.05) than healthy controls. In response to SEB, more CD107a-expressing CD8 + T cells were present in cases than controls (p<0.01). Despite differences in PBMC composition, PBMC from otitis-prone children mounted innate and T cell-mediated responses to NTHi challenge that were comparable to healthy children. These data provide evidence that otitis-prone children do not have impaired functional cell mediated immunity.

Research paper thumbnail of Analysis by proteomics reveals unique circulatory proteins in idiopathic pulmonary fibrosis

Respirology

Background and objective: Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic disease t... more Background and objective: Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic disease that has a poor 3-year median survival rate with unclear pathophysiology. Radiological features include bibasal, subpleural fibrosis and honeycombing while its pathology is characterized by fibroblastic foci and honeycombing. Proteomic analysis of circulating molecules in plasma may identify factors that characterize IPF and may assist in the diagnosis, prognostication and determination of pathogenic pathways in this condition. Methods: Two independent quantitative proteomic techniques were used, isobaric tags for relative and absolute quantitation (iTRAQ) and multiple reaction monitoring (MRM), to identify differentially expressed plasma proteins in a group of IPF patients in comparison to healthy controls with normal lung function matched for age and gender. Results: Five proteins were identified to be differentially expressed in IPF compared to healthy controls (upregulation of platelet basic protein and downregulation of actin, cytoplasmic 2, antithrombin-III, extracellular matrix protein-1 and fibronectin). Conclusion: This study further validates the combinational use of non-targeted discovery proteomics (iTRAQ) with targeted quantitation by mass spectrometry (MRM) of soluble biomarkers to identify potentially important molecules and pathways for pulmonary diseases such as IPF.

Research paper thumbnail of Circulating RNA differences between patients with stable and progressive lung function in IPF

European Respiratory Journal