Characteristics for ILD, COPD and healthy controls before and after propensity matching analysis.

Characteristics for ILD, COPD and healthy controls before and after propensity matching analysis.</p

Increased Synapse Formation Obtained by T Cell Epitopes Containing a CxxC Motif in Flanking Residues Convert CD4+ T Cells into Cytolytic Effectors

<div><p>The nature of MHC class II-binding epitopes not only determines the specificity of T cell responses, but may also alter effector cell functions. Cytolytic CD4+ T cells have been observed primarily in anti-viral responses, but very little is known about the conditions under which they can be elicited. Their potential as regulators of immune responses, however, deserves investigations. We describe here that inclusion of a thiol-disulfide oxidoreductase motif within flanking residues of class II-restricted epitopes results, both in vitro and in vivo, in elicitation of antigen-specific cytolytic CD4+ T cells through increased synapse formation. We show that both naïve and polarized CD4+ T cells, including Th17 cells, can be converted by cognate recognition of such modified epitopes. Cytolytic CD4+ T cells induce apoptosis on APCs by Fas-FasL interaction. These findings potentially open the way towards a novel form of antigen-specific immunosuppression.</p> </div

Patient characterization after propensity matching analysis.

Patient characterization after propensity matching analysis.</p

Diagnosis of ILD patients, n (%).

IIP: idiopathic interstitial pneumonias; IPF: idiopathic pulmonary fibrosis; INSIP: idiopathic nonspecific interstitial pneumonia; COP: cryptogenic organizing pneumonia; IIP: idiopathic interstitial pneumonias; EAA: extrinsic allergic alveolitis; CTD-ILD: connective tissue disease-associated interstitial lung disease. (DOCX)</p

Correlations between daily steps and clinical characteristics.

BMI: body mass index; FEV1: forced expiratory volume in the first second; FVC: forced vital capacity; DLCO: diffusion capacity for carbon monoxide; 6MWD: six-minute walking distance; QF: quadriceps force. r: Pearson correlation coefficient; p: p-value. (DOCX)</p

Effects of aminoacid residue mutation in the p21–35 flanking sequence.

<p>(A) Proliferation response and cytolytic properties of a p21–35 specific CD4+ T cell clone (G121) obtained from peptide-immunized BALB/c mice. Various mutant peptides (0.1 µM; sequence as indicated) were cultured with mitomycin C-treated splenocytes (T cell depleted). After 48 h, ³H-thymidine was added and its incorporation measured after 18 h of culture. Results are shown as percentage of incorporation obtained by comparison with p21–35 wildtype sequence (containing a CxxS motif). G121-induced cytolysis was measured on Dioc18-stained WEHI 231 B cells loaded with 0.1 µM of each peptide. After 24 h of incubation, cells were stained with Annexin V and 7-AAD and analysed by flow-cytometry. WEHI cell lysis (Dioc18+) is represented as percentage of lysis obtained by comparison with p21–35 wildtype sequence. Error bars represent 1 standard deviation (SD). Data are representative of three independent experiments. (B) Cytolytic assay of WEHI 231 B cells carried out as in (A) using two different CD4+ clones (G121 and R3TB7) exposed to peptides in either p21–35 wildtype sequence or after introduction of a cysteine in P(−1) thereby creating a thio-oxidoreductase motif. Error bars represent 1 SD. Data representative of two experiments. (C) Conjugate formation. DioC18 stained WEHI 231 B cells were loaded for 1 h with 1 µM of either Der p 2 p21–35, mutant peptide P(−1)Ala or with peptide containing the sequence CxxCGG. A CellTrace DDAO stained p21–35 specific clone (R3TB7) was added to each WEHI cell culture (ratio 1/1) and cell mixtures were briefly centrifuged. After 15 or 30 minutes, cells were softly resuspended, fixed for 10 minutes with 4% paraformaldhyde and analyzed by flow cytometry. Percentages of conjugates stand for the proportion of T cells staining positive for both labels over the total T cell population. Error bars represent 1 SD. Two tailed <i>p</i> values are derived from unpaired t tests. Data representative of two experiments. (D) Catabolism of CD3-ζ. On day 2 after restimulation, a p21–35 specific clone R3TB6 was washed and cultured for 15 minutes with WEHI 231 B cells either unloaded or preloaded with p21–35, CxxCGGp21–35 or loss of function peptide AxxAGGp21–35 (50 µM each). After fixation and permeation, expression of phosphorylated CD3-ζ chain was determined by flow cytometry. Data representative of three experiments and different p21–35 specific cell lines. (E) The same cells as in (D) were stained for surface CD3-ε or TCR-β after 1 h of culture with WEHI 231 B cells either unloaded or preloaded with p21–35, CxxCGGp21–35 or loss of function peptide AxxAGGp21–35 (50 µM each). Data representative of two experiments.</p

Absolute and relative active time per day in patients with ILD, COPD and healthy controls.

Distribution of active periods (METs level ≥1.5) in a day, expressed as the amount of active minutes per two hours (panel A) and the relative distribution of a patient’s active time over the day during waking hours (7AM to 9PM) (panel B) in patients with ILD, COPD and healthy controls.</p

Example of a physical activity report of one day.

Example of a physical activity report of one day, measured by SenseWear Pro armband. Mean intensity (METs) during waking hours: mean energy expenditure expressed in metabolic equivalent of a task between 7AM until 10PM. Mean intensity (METs) when active: mean energy expenditure expressed in METs when METs level of 1.5 is exceeded. MVPA bout ≥2’: period of at least two minutes when METs level ≥3. Total time MVPA per day is the summation of these periods in one day. MVPA bout ≥2’ is also used to calculate the mean duration of a MVPA bout. MVPA bout ≥10’: period of at least 10 minutes when METS level ≥3.</p

Characteristics for ILD group for matched and unmatched patients.

Results shown as mean ± standard error of the mean. M/F: male/female; %M: percentage of males in the sample; BMI: body mass index; 6MWD: six-minute walking distance. P-value for comparison between ILD matched and unmatched population. (DOCX)</p

Correlation between functional exercise capacity and physical activity in patients with ILD, COPD and healthy controls.

Correlation between six-minute walking distance (x-axis) and daily steps (y-axis) in patients with ILD (black dots), COPD (grey dots) and healthy controls (open squares). *P<0.05.</p