Immunosuppressive treatment helps to reduce the risk of rejection after transplantation. However, it is important to be aware of the potential toxicities that can develop after this therapy initiation. Tacrolimus is currently the calcineurin inhibitor of choice in the post-transplantation immunosuppressive therapy, and its intra-patient variability (Tac-IPV) has been proposed in recent years as a useful biomarker in post-transplant outcomes. We performed an observational retrospective study in 195 lung transplant recipients to determine the potential impact of Tac-IPV as a biomarker in the development of cardiovascular toxicities and renal impairment in the post-transplantation evolution. Different Tac-IPV calculation measurements were shown to be statistically associated with the development of arterial hypertension de novo in the univariate analysis: the standard deviation (p = 0.016 for tacrolimus Cmin values; p = 0.022 for tacrolimus dose-adjusted values); the coefficient of variation (p = 0.021 for Cmin; p = 0.031 for dose-adjusted values); and the mean absolute deviation (MAD) (p = 0.043 for Cmin; p = 0.033 for dose-adjusted values). Those patients with a MAD ≥ 16.47% had a higher risk of renal impairment (p = 0.045). In our study, different Tac-IPV measures were associated with arterial hypertension in lung transplantation follow-up. Patients with a high Tac-IPV, using the MAD as a measurement, showed a higher risk for renal impairment. No other statistically significant associations were found for the other studied events.

Background/Objectives: Patients with rheumatic immune-mediated inflammatory diseases (R-IMID) require latent tuberculosis infection screening and, in case of positivity, chemoprophylaxis. Isoniazid INH remains the standard regimen, but hepatotoxicity is an underrecognized concern. To describe the characteristics of R-IMID patients developing hepatotoxicity during INH therapy and identify potential risk factors through clinical analysis and literature review. Methods: Retrospective study of 64 R-IMID who developed hepatotoxicity with INH. Mean age was 53.4 ± 10.5 years; 70.3% female. Diagnoses included spondyloarthritis/psoriatic arthritis (56.3%), rheumatoid arthritis (32.8%), systemic sclerosis (4.7%), connective tissue diseases (4.7%), and other IMIDs (3.2%). All patients showed ≥ 2 × upper limit of normality (ULN) liver enzyme elevation, 34.4% ≥ 3 ULN, 20.3% ≥ 4 ULN. Literature review (19 studies) revealed INH-related hepatotoxicity rates of 1-41%, exacerbated by concurrent methotrexate, sulfasalazine, TNF inhibitors, and prior drug-induced liver injury. Results: Hepatotoxicity was frequent when INH was combined with other hepatotoxic drugs, especially methotrexate. Conclusions: INH prophylaxis in R-IMID patients carries substantial hepatotoxic risk. Careful hepatic monitoring and individualized risk stratification are essential to prevent liver injury in immunosuppressed populations.

Inflammatory cell activation in asthma may lead to reactive oxygen species (ROS) overproduction with an imbalance between oxidant levels and antioxidant capacity, called oxidative stress (OS). Since particulate matter (PM) airborne exposure may also contribute to ROS generation, it is unclear whether PM contributes more to OS than inflammatory cell activation. In our ASTHMA-FENOP study, which included 44 asthma patients and 37 matched controls, we aimed to characterize OS using five serum markers: total ROS content, protein carbonyl content, oxidized low-density lipoprotein (OxLDL), 8-hydroxydeoxyguanosine, and glutathione. Volunteers wore personal samplers for 24 h, collecting fine and coarse PM fractions separately, and the oxidative potential (OP) was determined using two methods. We observed differences between asthmatic and non-asthmatic volunteers in some OS markers, such as OxLDL, with an adjusted mean difference of 50,059.8 ng/mL (p < 0.001). However, we did not find an association between higher PM-OP and increased systemic OS. This suggests that at our PM-OP exposure levels, OS generated by the inflammatory cells themselves is more relevant than that generated by airborne PM. This supports the idea that asthma is a heterogeneous disease at the molecular level, mediated by inflammatory cell activation, and that OS may have potential clinical implications.

Background. Antibody-mediated rejection (ABMR) has become one of the leading causes of chronic lung graft dysfunction. However, in lung transplantation, this entity is sometimes difficult and controversial to diagnose. It is mainly caused by the appearance of donor- specific anti-human leukocyte antigen (HLA) antibodies (DSA), although there are situations with Cod deposits in biopsy in the absence of circulating DSA. The aim of this work was to study the potential role of non-HLA antibodies in the development of ABMR without DSA after lung transplantation. Methods. A case-control study was designed with a cohort of lung transplant recipients at our institution. Twenty-seven patients with ABMR and without anti-HLA antibodies were identified after lung transplantation, and a control group of 21 transplant recipients was selected with the same post-transplant follow-up without evidence of rejection. Non-HLA antibodies were studied pretransplant using Luminex (ThermoFisher, One Lambda). Results. The median of the pretransplant non-HLA-positive antibodies in the group with ABMR without DSA is significantly higher than in the control group: 2 (interquartile range, 0-16) vs 0 (interquartile range, 0-1; P .01). Patients with 1.5 pretransplant non-HLA antibodies were more likely to develop ABMR without DSA (sensitivity, 80.95%; specificity, 55.55%; area under the curve, 71.3%). Conclusion. The increase of non-HLA antibodies before lung transplantation has recently been shown to increase the risk of chronic lung allograft dysfunction. These results confirm that patients with a higher number of non-HLA antibodies could be at risk of developing ABMR without DSA. These results point out the possible usefulness of pre-lung transplant non-HLA antibodies to identify patients with end-stage lung disease at risk of developing ABMR without DSA.

To improve lung transplant recipient (LungTx) outcome, it would be of great interest to measure the net state of immunosuppression to avoid both infection and rejection. Measurement of Torquetenovirus load (TTV load) has been proposed as a biomarker to monitor solid organ transplantation, but its relationship with immunosuppressive drugs, particularly mycophenolic acid (MPA), is not well understood. We performed a prospective study of 53 LungTx, measuring TTV load before transplantation, at week 3, and at month 3. Tacrolimus and MPA doses and levels were recorded, and an area under the MPA curve (AUC-MPA) was calculated at the third month. LungTx in the fourth quartile of TTV load at the third week and the third month exhibited a low risk of acute rejection (OR 0.113, 95% CI 0.013-0.953, p = 0.045) and a high risk of opportunistic infection from month 3 to 6 (OR 15.200, 95% CI 1.525-151.511, p = 0.020), respectively. TTV load was weakly related to tacrolimus trough level at month 3 (rho = 0.283, p = 0.040). Neither MPA blood levels nor AUC-MPA were related to TTV load, although only patients with a reduction in MPA dose from month 1 to 3 showed a smaller increase in TTV load (0.86, IQR 2.58 log10 copies/mL vs. 2.26, IQR 3.02 log10 copies/mL, p = 0.026). In conclusion, TTV load in LungTx is only partially related to exposure to immunosuppressive drugs. Other variables, such as inflammation, immunosenescence, and frailty, may influence the overall level of immunosuppression and TTV load.

Background/Objectives: Some lung transplant (LungTx) recipients do not achieve the expected lung function within the first year, a condition known as baseline lung allograft dysfunction (BLAD). Our objective was to analyze the risk factors associated with BLAD, focusing on the variables associated with a higher risk of developing a more intense alloimmune response. Methods: We carried out a prospective study including 88 LungTx recipients. BLAD was defined as failure to reach 80% of the predicted value for forced expiratory volume in one second (FEV1) and/or forced vital capacity (FVC) on two tests conducted at least three weeks apart. Tacrolimus time in therapeutic range (TTR) and mycophenolic acid area under the curve (MPA AUC0–12h) were measured at the third month. Donor–recipient compatibility was assessed using HLA eplet mismatch analysis, performed via HLA Matchmaker 3.1. Results: BLAD patients showed greater eplet mismatch burden (67, IQR 20 vs. 55, IQR 22, p = 0.018) and had been exposed to a lower TTR (26.6%, IQR 14.0% vs. 39.6%, IQR 24.3%, p = 0.039) and less frequently to an adequate third-month MPA AUC0–12 > 30 mg × h/L (57.1% vs. 89.2%, p = 0.020). DR/DQ eplet mismatches (β = −0.348, p = 0.002) and third-month MPA AUC0–12 (β = 0.285, p = 0.009) were independently associated with six-month predicted FEV1%. Conclusions: Among other variables, BLAD and initial lung graft function are associated with greater eplet discordance and lower immunosuppressive drug exposure, suggesting a potential role of underlying alloimmune responses in their pathogenesis.

With the rationale that the oxidative potential of particulate matter (PM-OP) may induce oxidative stress and inflammation, we conducted the ASTHMA-FENOP study in which 44 asthmatic patients and 37 matched controls wore a personal sampler for 24 h, allowing the collection of fine and coarse PM fractions separately, to determine PM-OP by the dithiothreitol (DTT) and ascorbic acid (AA) methods. The levels of Interleukin 6 (IL-6) and the IL-6/IL-10 ratio, as indicators of pro- and anti-inflammatory statuses, were determined by calculating the mean differences (MDs), odds ratios (ORs) and p-trends adjusted for sex, age, study level and body mass index. Positive associations for IL-6 levels in the form of adjusted MDs and ORs were obtained for all PM-OP metrics, reaching statistical significance for both OP-DTT and OP-AA in the fine fraction, with adjusted OR = 5.66; 95%CI (1.46 to 21.92) and 3.32; 95%CI (1.07 to 10.35), respectively, along with statistically significant dose-response patterns when restricting to asthma and adjusted also for clinical variables (adjusted p-trend = 0.029 and 0.01). Similar or stronger associations and dose-response patterns were found for the IL-6/IL-10 ratio. In conclusion, our findings on the effect of PM-OP on systemic inflammation support that asthma is a heterogeneous disease at the molecular level, with PM-OP potentially playing an important role.

Background/Objectives: Patients with rheumatic immune-mediated diseases (rheumatic-IMID) and latent tuberculosis (LTBI) are at an increased risk of developing active tuberculosis (TB); therefore, screening is recommended before starting biological treatment. The aims of this study were as follows: (i) to assess the prevalence of LTBI, (ii) to determine the importance of using a booster test in TST-negative patients, (iii) to compare the tuberculin skin test (TST) with the interferon-gamma release assay (IGRA), (iv) to perform a review of the prevalence of LTBI. Methods: A cross-sectional hospital study was performed, including patients diagnosed with rheumatic-IMID who underwent a TST and/or IGRA during the period 2016-2020. If the first TST was negative, a new TST (booster) was performed. Results: A total of 1117 patients were included. The overall prevalence of LTBI was estimated to be 31.7% (95% confidence interval, 29.74-33.66). The LTBI prevalence ranged from 38.5% for vasculitis to 14% for sarcoidosis. The booster test was positive in 22.9% of 817 patients with a negative or indeterminate IGRA. The IGRA was positive in 3.8% of 793 patients with a negative booster.The adjusted Cohen's kappa coefficient between TST (+booster) and IGRA was 0.62. Conclusions: LTBI is frequent in patients with rheumatic-IMID. IGRA and TST (+booster) show a moderate, fair grade of agreement. Therefore, performing both tests before biological therapy should be highly recommended.

The most critical forms of coronavirus disease 2019 (COVID-19) are associated with excessive activation of the inflammasome. Despite the COVID-19 impact on public health, we still do not fully understand the mechanisms by which the inflammatory response influences disease prognosis. Accordingly, we aimed to elucidate the role of polymorphisms in the key genes of the formation and signaling of the inflammasome as biomarkers of COVID-19 severity. For this purpose, a large and well-defined cohort of 377 COVID-19 patients with mild (n = 72), moderate (n = 84), severe (n = 100), and critical (n = 121) infections were included. A total of 24 polymorphisms located in inflammasome-related genes (NLRP3, NLRC4, NLRP1, CARD8, CASP1, IL1B, IL18, NFKB1, ATG16L1, and MIF) were genotyped in all of the patients and in the 192 healthy controls (HCs) (who were without COVID-19 at the time of and before the study) by RT-qPCR. Our results showed that patients with mild, moderate, severe, and critical COVID-19 presented similar allelic and genotypic distribution in all the variants studied. No statistically significant differences in the haplotypic distribution of NLRP3, NLRC4, NLRP1, CARD8, CASP1, IL1B, and ATG16L1 were observed between COVID-19 patients, who were stratified by disease severity. Each stratified group of patients presented a similar genetic distribution to the HCs. In conclusion, our results suggest that the inflammasome polymorphisms studied are not associated with the worsening of COVID-19.

A simplified 4-strata risk stratification approach based on three variables is widespread in pulmonary arterial hypertension (PAH) at follow-up. This study aimed to assess the impact of replacing the 6-min walk test (6MWT) with the peak 02 uptake evaluated by the cardiopulmonary exercise test (CPET) on risk stratification by this scale. We included 180 prevalent patients with PAH from two reference hospitals in Spain, followed up between 2006 and 2022. Patients were included if all the variables of interest were available within a 3-month period on the Spanish Registry of Pulmonary Arterial Hypertension (REHAP): functional class (FC); NT-proBNP; 6MWT; and CPET. The original 4-strata model (NT-proBNP, 6MWT, FC) identified most patients at low or intermediate-low risk (36.7% and 51.1%, respectively). Notably, the modified scale (NT-proBNP, CPET, FC) improved the identification of patients at intermediate-high risk up to 18.9%, and at high risk up to 1.1% in comparison with the previous 12.2% and 0.0% in the original scale. This new model increased the number of patients correctly classified into higher-risk strata (positive NRI of 0.06), as well as classified more patients without events in lower-risk strata (negative NRI of 0.04). The proposed score showed a slightly superior prognostic capacity compared with the original model (Harrel's C-index 0.717 vs. 0.709). Using O2 uptake instead of distance walked in the 6MWT improves the identification of high-risk patients using the 4-strata scale. This change could have relevant prognostic implications and lead to changes in the specific treatment of PAH.

Our study aims to evaluate the effect of everolimus treatment on lung function in lung transplant (LT) patients with established chronic lung allograft dysfunction (CLAD). Methods: This retrospective study included LT patients in two reference LT units who started everolimus therapy to treat CLAD from October 2008 to October 2016. We assessed the variation in the maximum forced expiratory volume in the first second (FEV1) before and after the treatment. Results: Fifty-seven patients were included in this study. The variation in the FEV1 was −102.7 (149.6) mL/month before starting everolimus compared to −44.7 (109.6) mL/month within the first three months, +1.4 (63.5) mL/month until the sixth month, and −7.4 (46.2) mL/month until the twelfth month (p < 0.05). Glomerular filtrate remained unchanged after everolimus treatment [59.1 (17.5) mL/min per 1.73 m2 at baseline and 60.9 (19.6) mL/min per 1.73 m2, 57.7 (20.5) mL/min per 1.73 m2, and 57.3 (17.8) mL/min per 1.73 m2, at 1, 3, and 6 months, respectively] (p > 0.05). Everolimus was withdrawn in 22 (38.6%) patients. The median time to withdrawal was 14.1 (5.5–25.1) months. Conclusions: This study showed an improvement in FEV1 decline in patients with CLAD treated with everolimus. However, the drug was withdrawn in a high proportion of patients.

Objective The CLASS (Classification Criteria of Anti-Synthetase Syndrome) project is a large international multicentre study that aims to create the first data-driven anti-synthetase syndrome (ASSD) classification criteria. Identifying anti-aminoacyl tRNA synthetase antibodies (anti-ARS) is crucial for diagnosis, and several commercial immunoassays are now available for this purpose. However, using these assays risks yielding false-positive or false-negative results, potentially leading to misdiagnosis. The established reference standard for detecting anti-ARS is immunoprecipitation (IP), typically employed in research rather than routine autoantibody testing. We gathered samples from participating centers and results from local anti-ARS testing. As an "ad-interim" study within the CLASS project, we aimed to assess how local immunoassays perform in real-world settings compared to our central definition of anti-ARS positivity. Methods We collected 787 serum samples from participating centres for the CLASS project and their local anti-ARS test results. These samples underwent initial central testing using RNA-IP. Following this, the specificity of ARS was reconfirmed centrally through ELISA, line-blot assay (LIA), and, in cases of conflicting results, protein-IP. The sensitivity, specificity, positive likelihood ratio and positive and negative predictive values were evaluated. We also calculated the inter-rater agreement between central and local results using a weighted kappa co-efficient. Results Our analysis demonstrates that local, real-world detection of anti-Jo] is reliable with high sensitivity and specificity with a very good level of agreement with our central definition of anti-Jo] antibody positivity. However, the agreement between local immunoassay and central determination of anti-non-Jo] antibodies varied, especially among results obtained using local LIA, ELISA and "other" methods. Conclusion Our study evaluates the performance of real-world identification of anti-synthetase antibodies in a large cohort of multi-national patients with ASSD and controls. Our analysis reinforces the reliability of real-world anti-Jo] detection methods. In contrast, challenges persist for anti-non-Jo] identification, particularly anti-PL7 and rarer antibodies such as anti-OJ/KS. Clinicians should exercise caution when interpreting anti-synthetase antibodies, especially when commercial immunoassays test positive for non-anti-Jo] antibodies.

Background With no approved therapies for pulmonary hypertension (PH) associated with interstitial lung disease (PH-ILD) in Europe, we surveyed clinician perceptions on PH-ILD management and unmet need to understand current real-world practices. Methods An online clinician survey on PH-ILD management was conducted in France, Germany, Italy, Spain and the UK. Results 55 clinicians (78% pulmonologists), each managing a median 20 PH-ILD patients (interquartile range (IQR) 10–50), participated. Upon PH suspicion, clinicians referred a median 50% (IQR 20–73%) of patients for echocardiography alone and 35% (IQR 20–78%) for echocardiography, followed by right heart catheterisation. Upon diagnosis, a median 20% (IQR 9–30%), 40% (IQR 20–50%) and 35% (IQR 20–55%) of patients fell under the pulmonary arterial pressure ranges of 21–24 mmHg, 25–34 mmHg and >35 mmHg, respectively. 50% of patients received off-label treatment for their PH and, of those, off-label phosphodiesterase-5 inhibitor (PDE-5i), endothelin receptor antagonist (ERA) and prostacyclin analogues were prescribed first-line by 78%, 9% and 7% of clinicians, respectively. Upon PDE-5i non-response, 35% of clinicians proceed with an ERA, 35% with no further therapy. 55% of clinicians used dual-therapy. Yearly median inpatient admissions and emergency visits were 2.0 (IQR 1.3–2.9) and 1.5 (IQR 1.0–2.0), respectively (n=31 responses). Most clinicians (69%) highlighted lack of efficacy or evidence for current therapies as a key gap in PH-ILD management. Conclusions This study gives insight into real-world European PH-ILD diagnosis and management. With significant use of off-label treatment, there is a large unmet need due to lack of approved therapies. Despite updated guidelines, more evidence is needed to standardise PH-ILD management.