Extracellular heat shock protein 70 (eHsp70) might modulate immune responses in chronic obstructive pulmonary disease (COPD). The aim of the study was to explore eHsp70 concentration in stable COPD, its association with disease severity and smoking status as well as its diagnostic performance in COPD assessment. Plasma samples were collected from 137 COPD patients and 95 healthy individuals, and concentration of eHsp70 was assessed by commercially available enzyme-linked immunosorbent assay (ELISA) kit (Enzo Life Science, Farmingdale, NY, USA). COPD patients were subdivided regarding airflow obstruction severity and symptoms severity according to the Global Initiative for COPD (GOLD) guidelines. eHsp70 concentration increased in COPD patients when compared to controls and increased with the severity of airflow limitation as well as symptoms burden and exacerbation history. eHsp70 concentration did not differ among COPD patients based on smoking status, yet it increased in healthy smokers compared to healthy nonsmokers. In addition, eHsp70 negatively correlated with lung function parameters forced expiratory volume in one second (FEV₁) and FEV₁/ forced vital capacity (FVC), and positively with COPD multicomponent indices BODCAT (BMI, airflow obstruction, dyspnea, CAT score), BODEx (BMI, airflow obstruction, dyspnea, previous exacerbations), CODEx (Charlson’s comorbidity index, airflow obstruction, dyspnea, previous exacerbations) and DOSE (dyspnea, airflow obstruction, smoking status, previous exacerbations) With great predictive value (OR = 7.63) obtained from univariate logistic regression, eHsp70 correctly classified 76% of cases. eHsp70 is associated with COPD prediction and disease severity and might have the potential for becoming an additional biomarker in COPD assessment.

Introduction: Chronic obstructive pulmonary disease (COPD) is a condition with underlying systemic inflammation and haemostatic changes. Aims and Objectives: Assessment of the platelet indices - platelet count (PLT), mean platelet volume (MPV), platelet distribution width (PDW) and platelet to lymphocyte ratio (PLR), evaluation of their diagnostic value and association with well-known inflammatory markers. Methods: 109 patients with stable COPD and 95 healthy controls were included. Blood samples were collected in different tubes depending on the analysis. The results were analyzed by Mann-Whitney Rank Sum test performed by MedCalc statistical software. Results: Among platelet indices, PLT (P=0.007) and PLR (P=0.005) were significantly increased, while MPV (P<0.001) and PDW (P<0.001) were decreased in COPD patients. Inflammatory markers, CRP (P<0.001), fibrinogen (P<0.001) and leukocyte count (P<0.001) were increased in COPD patients. Positive correlation between fibrinogen and platelet count was observed (r=0.396, P<0.001). Moreover, fibrinogen showed to be a good diagnostic predictor of COPD (OR=2.5529, P<0.001). Leukocyte count, PDW and PLR correctly classified 75% of COPD cases with an AUC of 0.845 in a multivariate logistic regression analysis. Conclusions: Our study suggests that thrombocytosis is present in chronic inflammation observed in COPD patients. Fibrinogen may be useful as a COPD predictor. Due to its association with PLT, it confirmed systemic inflammation and haemostatic changes in COPD. Moreover, PLT showed a good performance as a COPD predictor when combined with lymphocytes in a ratio. Model combining PLR, PDW and leukocyte count has good discrimination power among healthy subjects and COPD patients.

Heat-shock protein 70 (Hsp70) acts as a damage-associated molecular pattern (DAMP) in extracellular compartment and is capable to induce secretion of pro-inflammatory cytokines and trigger innate immunity. The aim of the study was to assess whether its concentration is increased in plasma of patients with stable chronic obstructive pulmonary disease (COPD) and to explore its association with smoking status, severity of airflow obstruction, symptoms and exacerbations history. Blood samples were collected from 137 COPD patients and 95 healthy volunteers matched by age and sex. COPD patients were subdivided based on airflow obstruction severity (stages 1-4) and symptoms severity (ABCD assessment) as recommended by Global Initiative for Chronic Obstructive Lung Disease (GOLD). Concentration of extracellular Hsp70 (eHsp70) was measured by the commercially available ELISA kit. Concentration of eHsp70 was increased in COPD patients in comparison to control group (P<0.001). It was also increasing with the severity of airflow obstruction throughout GOLD 2-4 stages (P<0.001) as well as with symptoms severity and exacerbations frequency throughout GOLD A-D groups (P<0.001). There were no differences in eHsp70 concentrations among COPD patients based on smoking status (non-smokers, former smokers, smokers), yet eHsp70 was increased in healthy smokers compared to healthy non-smokers (P<0.001). Moreover, eHsp70 showed significant negative correlation with FEV1 (r=-0.708, P<0.001) and FEV1/FVC (r=-0.644, P<0.001), while its odds ratio (OR) was 7.63 (95% CI = 3.68 – 15.82, P<0.001). eHsp70 may contribute to the inflammation in COPD associated with disease progression. Therefore, it could become one of the prognostic biomarkers for COPD severity.

Complete molecular mechanisms that underlie inflammatory responses in chronic obstructive pulmonary disease (COPD) are still unknown but seem to be associated with chronic exposure to noxious particles or gases, particularly cigarette smoke, and genetic susceptibility. The aim of this study was to explore gene expressions of HSP70 as well as TLR2 and TLR4 (the main receptors for extracellular Hsp70), and association of their polymorphisms with risk of COPD. 137 patients with stable COPD and 95 controls were included in the study. Real-time polymerase chain-reaction was used for the determination of HSP70, TLR2 and TLR4 mRNA expressions as well as for genotyping for single nucleotide polymorphisms (SNPs) of HSP70 (rs1008438, rs6457452), TLR2 (rs1898830, rs3804099, rs13150331) and TLR4 (rs2737190, rs10759932, rs7846989) genes. Levels of HSP70 (P<0.001) and TLR2 (P=0.002) mRNA expression were increased in COPD patients compared to healthy subjects and were also increased at each disease stage regarding the airway obstruction (GOLD 2-4 with P<0.001, P=0.019, respectively) as well as in each group based on symptoms severity assessed by CAT and history of exacerbations (GOLD A-D with P<0.001, P=0.009, respectively) compared to controls. TLR4 gene expression was similar in controls and COPD patients. After smoking adjustments, A/C genotype of SNP rs1008438 of HSP70 gene showed to be associated with a decreased risk of COPD in Croatian population in overdominant model with OR (95% CI) of 0.52 (0.28-0.96) (P=0.036). Enhanced gene expression of HSP70 and TLR2 might be involved in immune response in stable COPD, while HSP70 rs1008438 variation might play important role in the genetic susceptibility to COPD.

Heat-shock protein 70 (Hsp70) acts as a damage-associated molecular pattern (DAMP) in extracellular compartment and might induce secretion of pro-inflammatory cytokines and trigger innate immunity. The aim of the study was to assess whether its concentration is increased in plasma of patients with stable chronic obstructive pulmonary disease (COPD) and to explore its association with smoking status, severity of airflow obstruction, symptoms and exacerbations history. Blood samples were collected from 137 COPD patients and 95 healthy volunteers matched by age and sex. COPD patients were subdivided based on FEV₁-based severity (stages 1-4) and symptoms severity (ABCD assessment), as recommended by Global Initiative for Chronic Obstructive Lung Disease (GOLD). Concentration of extracellular Hsp70 (eHsp70) was measured by the commercially available ELISA kit. Concentration of eHsp70 was increased in COPD patients in comparison to control group (p<0.001). It was also increasing with the severity of airflow obstruction throughout GOLD 2-4 stages (P<0.001) as well as with symptoms severity and exacerbations frequency throughout GOLD A-D groups (P<0.001). There were no differences in eHsp70 concentrations among COPD patients based on smoking status (non-smokers, former smokers, smokers), yet eHsp70 was increased in healthy smokers compared to healthy non-smokers (P<0.05). Moreover, eHsp70 showed significant negative correlation with FEV₁ (r=-0.708, p<0.001) and FEV₁/FVC (r=-0.644, P<0.001), while its odds ratio (OR) was 7.63 (95% CI = 3.68 – 15.82, P<0.001). eHsp70 may contribute to COPD inflammation associated with disease progression and could become prognostic biomarker for disease severity.