Evaluation of the relationship between surfactant protein D levels and COVID-19 clinical severity: a case-control study

Purpose: Serum surfactant protein D (SP-D) plays roles in the body such as protection against viral infection, bacterial and fungal clearance, clearance of apoptotic cells and suppression of inflammation. This study aims to examine the relationship between SP-D level and coronavirus disease (COVID-19) severity. Methods: 80 patients (30 with mild disease and 50 with severe/critical COVID-19), and 50 healthy volunteers were enrolled in the study. SP-D levels were analyzed by ELISA in serum samples. Results: The median of SP-D was found to be 2.47 (1.67-7.79) ng/ml in mild disease and 5.65 (3.09-16.55) ng/ ml in severe/critical disease groups, while 2.89 (10.8-6.24) ng/ml in the healthy controls. The differences in SP-D levels between the severe/critical disease group compared to both mild disease and control groups were found statistically significant (p=0.007 and 0.001, respectively). ROC analysis showed greater AUC for the serum SP-D levels of the severe/critical COVID-19 patients compared to mild COVID-19 disease patients (AUC=0,691, 95% CI=0.56-0,822;p=0.004). Furthermore, SP-D levels were 86% sensitive and 51.6% specific at 2.44 ng/ml level (p=0.004) to detect severe/critical patients. Conclusion: SP-D levels is useful for COVID-19 patients in the prediction of clinical severity and prognosis. SP-D is a valuable biomarker for predicting the clinical severity and prognosis. © 2023, Pamukkale University. All rights reserved.

Malondialdehyde acetaldehyde adduction of surfactant protein D attenuates SARS-CoV-2 spike protein binding and virus neutralization.

BACKGROUND: Over 43% of the world's population regularly consumes alcohol. Although not commonly known, alcohol can have a significant impact on the respiratory environment. Living in the time of the COVID-19 pandemic, alcohol misuse can have a particularly deleterious effect on SARS-CoV-2-infected individuals and, in turn, the overall healthcare system. Patients with alcohol use disorders have higher odds of COVID-19-associated hospitalization and mortality. Even though the detrimental role of alcohol on COVID-19 outcomes has been established, the underlying mechanisms are yet to be fully understood. Alcohol misuse has been shown to induce oxidative damage in the lungs through the production of reactive aldehydes such as malondialdehyde and acetaldehyde (MAA). MAA can then form adducts with proteins, altering their structure and function. One such protein is surfactant protein D (SPD), which plays an important role in innate immunity against pathogens. METHODS AND RESULTS: In this study, we examined whether MAA adduction of SPD (SPD-MAA) attenuates the ability of SPD to bind SARS-CoV-2 spike protein, reversing SPD-mediated virus neutralization. Using ELISA, we show that SPD-MAA is unable to competitively bind spike protein and prevent ACE2 receptor binding. Similarly, SPD-MAA fails to inhibit entry of wild-type SARS-CoV-2 virus into Calu-3 cells, a lung epithelial cell line, as well as ciliated primary human bronchial epithelial cells isolated from healthy individuals. CONCLUSIONS: Overall, MAA adduction of SPD, a consequence of alcohol overconsumption, represents one mechanism of compromised lung innate defense against SARS-CoV-2, highlighting a possible mechanism underlying COVID-19 severity and related mortality in patients who misuse alcohol.

Labeled TEMPO-Oxidized Mannan Differentiates Binding Profiles within the Collectin Families.

Establishing the rapid and accurate diagnosis of sepsis is a key component to the improvement of clinical outcomes. The ability of analytical platforms to rapidly detect pathogen-associated molecular patterns (PAMP) in blood could provide a powerful host-independent biomarker of sepsis. A novel concept was investigated based on the idea that a pre-bound and fluorescent ligand could be released from lectins in contact with high-affinity ligands (such as PAMPs). To create fluorescent ligands with precise avidity, the kinetically followed TEMPO oxidation of yeast mannan and carbodiimide coupling were used. The chemical modifications led to decreases in avidity between mannan and human collectins, such as the mannan-binding lectin (MBL) and human surfactant protein D (SP-D), but not in porcine SP-D. Despite this effect, these fluorescent derivatives were captured by human lectins using highly concentrated solutions. The resulting fluorescent beads were exposed to different solutions, and the results showed that displacements occur in contact with higher affinity ligands, proving that two-stage competition processes can occur in collectin carbohydrate recognition mechanisms. Moreover, the fluorescence loss depends on the discrepancy between the respective avidities of the recognized ligand and the fluorescent mannan. Chemically modulated fluorescent ligands associated with a diversity of collectins may lead to the creation of diagnostic tools suitable for multiplex array assays and the identification of high-avidity ligands.

Surfactant protein D (SP-D) as a biomarker of SARS-CoV-2 infection.

BACKGROUND: Surfactant protein-D (SP-D) is a lung-resident protein that has emerged as a potential biomarker for COVID-19. Previous investigations on acute respiratory distress syndrome patients demonstrated a significant increment of SP-D serum levels in pathological conditions. Since SP-D is not physiologically permeable to alveoli-capillary membrane and poorly expressed by other tissues, this enhancement is likely due to an impairment of the pulmonary barrier caused by prolonged inflammation. METHODS: A retrospective study on a relatively large cohort of patients of Hospital Pio XI of Desio was conducted to assess differences of the hematic SP-D concentrations among COVID-19 patients and healthy donors and if SP-D levels resulted a risk factor for disease severity and mortality. RESULTS: The first analysis, using an ANOVA-model, showed a significant difference in the mean of log SP-D levels between COVID-19 patients and healthy donors. Significant variations were also found between dead vs survived patients. Results confirm that SP-D concentrations were significantly higher for both hospitalized COVID-19 and dead patients, with threshold values of 150 and 250 ng/mL, respectively. Further analysis conducted with Logistic Mixed models, highlighted that higher SP-D levels at admission and increasing differences among follow-up and admission values resulted the strongest significant risk factors of mortality (model predictive accuracy, AUC = 0.844). CONCLUSIONS: The results indicate that SP-D can be a predictive marker of COVID-19 disease and its outcome. Considering its prognostic value in terms of mortality, the early detection of SP-D levels and its follow-up in hospitalized patients should be considered to direct the therapeutic intervention.

The role of surfactant proteins SP-A and SP-D in viral infection: a focus on COVID-19

An immune response to invasion of viral pathogens is an integral part of maintaining the physiological functioning of the bronchopulmonary system and effective gas exchange. Collagen-containing C-type lectins (lung collectins) are some of the key proteins in the identification of viral particles. They have image-recognizing receptors that identify pathogen-associated molecular patterns, particularly viral glycoproteins. The surfactant proteins SP-A and SP-D, which are composed of trimerized units, belong to pulmonary collectins and oligomerize into higher-order structures. These proteins play an essential role in recognition and elimination of microbial pathogens (viruses, bacteria, fungi, parasites, nanoparticles, allergens) through a variety of mechanisms. Taking into account the burden of the novel coronavirus infection caused by the SARS-CoV-2 virus, it is important to consider the role of the surfactant proteins SP-A and SP-D in the pathogenesis of the immune response to viral invasion. Currently, there are data on the direct relationship between surfactant proteins and viruses belonging to the Coronaviridae family. The SP-A and SP-D proteins modulate inflammatory responses and cytokine synthesis, but prevent an excessive inflammatory response (cytokine storm). There is also an assumption that SARSCoV-2 directly suppresses and alters the production of surfactant proteins. Thus, the key pathogenetic role of the surfactant proteins SP-A and SP-D in the response to the viral pathogen SARS-CoV-2 is evident. Today, this is a promising area of translational medicine, which will contribute to a profound understanding of the pathogenesis of coronavirus infection for assessing the diagnostic and prognostic potentials of the surfactant proteins SP-A and SP-D in COVID-19. Additionally, it will help evaluate the therapeutic potential of recombinant fragments of human SP-A and SP-D. © 2022 Siberian State Medical University. All rights reserved.

Elucidating the enhanced binding affinity of a double mutant SP-D with trimannose on the influenza A virus using molecular dynamics.

Surfactant protein D (SP-D) is an essential component of the human pulmonary surfactant system, which is crucial in the innate immune response against glycan-containing pathogens, including Influenza A viruses (IAV) and SARS-CoV-2. Previous studies have shown that wild-type (WT) SP-D can bind IAV but exhibits poor antiviral activities. However, a double mutant (DM) SP-D consisting of two point mutations (Asp325Ala and Arg343Val) inhibits IAV more potently. Presently, the structural mechanisms behind the point mutations' effects on SP-D's binding affinity with viral surface glycans are not fully understood. Here we use microsecond-scale, full-atomistic molecular dynamics (MD) simulations to understand the molecular mechanism of mutation-induced SP-D's higher antiviral activity. We find that the Asp325Ala mutation promotes a trimannose conformational change to a more stable state. Arg343Val increases the binding with trimannose by increasing the hydrogen bonding interaction with Glu333. Free energy perturbation (FEP) binding free energy calculations indicate that the Arg343Val mutation contributes more to the increase of SP-D's binding affinity with trimannose than Asp325Ala. This study provides a molecular-level exploration of how the two mutations increase SP-D binding affinity with trimannose, which is vital for further developing preventative strategies for related diseases.

Oleic acid-induced acute lung injury in Ob/Ob mice fed with high-fat-diet exacerbate bile acid uptake and liver injury that was reversed by antagonizing their entry

Background and aims: Acute respiratory distress syndrome (ARDS) is a serious complication of COVID-19 and present in a large percentage of COVID-19 deaths. Many studies suggest that people with obesity are at increased risk of severe COVID-19, however, mechanism on liver-lung axis remains unknown. We aimed to evaluate whether bile acid (BAs) trafficking interfere with acute lung injury (ALI) in animal model with obesity. Method: Leptin deficient (ob/ob) mice fed with high-fat-diet (Ob/Ob HFD) were i.p injected with oleic acid (OA) to induce ALI. To modulate BAs uptake, mice were i.p treated with neutralizing antibody for sodium taurocholate co-transporting polypeptide (NTCP;BAs-transporter). Broncho-alveolar lavage fluid (BALF), lungs, livers and serum were obtained from mice and assessed for inflammatory (HandE staining, ALT and pro-inflammatory panel of cytokines), fibrosis (Sirius red staining, a-smooth muscle actin, collagen and fibronectin) and metabolic (BAs, cholesterol, triglyceride, glucose tolerance test (GTT) and fasting blood sugar (FBS)) profiles. In addition, alveolarcapillary membrane injury of surfactant D (SP-D) and the receptor for advanced glycation end-products (RAGE). BAs trafficking were assessed in primary lung cells and their impact on proliferation and apoptosis were evaluated. Results: Compared to WT-littermates, OA-induced lung injury and was worsened in the in the Ob/Ob HFD in the histopathology outcome. In addition, BALF of the Ob/Ob HFD showed elevated levels of BAs (3- fold;P = 0.002) associated with increased GM-CSF, INF-g, IL-1, IL-6 and IL-8 (p < 0.01). Moreover, Ob/Ob HFD with OA showed elevated serum levels in liver enzymes, lipids, glucose and metabolic markers (p < 0.01). In addition, Ob/Ob HFD livers showed an exacerbated fibrosis profile. NTCP neutralizing antibody in Ob/Ob HFD while inhibited BAs uptake/trafficking in both primary alveolar type II (BALF showed 4-fold increase in BAs) and primary hepatocytes (serum showed 3-fold increase in BAs). SP-D, RAGE and serum metabolic markers were suppressed to normal in line with enhance lung and liver histology and maintaining cell viability. Conclusion: Modulation of BAs trafficking from the liver of obese mice to the lungs could be an important step in the pathogenesis of ALI. Antagonizing BAs uptake may suggest a therapeutic strategy in improving liver-lung axis.

Plasma and Airway Levels of Surfactant Protein D Degradation Reflect Progression of ARDS in Ventilated Patients with SARS-CoV-2

Background: Patients with COVID-19 present severe respiratory symptoms progressing to acute respiratory distress syndrome (ARDS). Upon infection, SARS-CoV-2 destroys cells expressing the ACE2 receptor including alveolar type II cells (AT2). These cells are found in the alveolar-capillary barrier which normally secrete pulmonary surfactant, a complex of lipid and surfactant proteins (SPA, SP-B, SP-C, SP-D). Exogenous surfactant therapy (mainly composed of phospholipids, SP-B, and SP-C) has been successful in treating neonatal respiratory distress syndrome (nRDS) caused by surfactant deficiency in preterm babies.Plasma SP-D has been proposed as a marker of lung injury in COVID-19 but so far, no reports have evaluated sequential SP-D levels in both airway and plasma. As part of a clinical trial repurposing surfactant therapy to treat adult ventilated COVID-19 patients, we hypothesized that plasma SP-D levels may reflect decreased lung integrity and that SP-D degradation in plasma and airway samples from COVID-19 patients may reflect disease progression and severity. Methods: Enzyme-linked immunosorbent assay (ELISA) was used to quantify SP-D concentration in patient plasma and tracheal aspirate samples. Western Blotting was used to identify any protein degradation. Sequential daily plasma and airway samples were analysed. Results: SP-D concentration in serum was 10-20 times higher in patients ventilated for COVID-19 than in healthy volunteers. Additionally, the concentration of SP-D in plasma has shown to be 10-100-fold higher than in tracheal aspirates. Furthermore, degraded fragments of SP-D were detected at a higher ratio than intact SP-D in plasma of ventilated patients. This ratio decreased with administration of surfactant therapy (containing phospholipids and SP-B and SP-C but no SPA or SP-D). Conclusions: Increased serum SP-D and decreased tracheal aspirate SP-D from ventilated COVID-19 patients suggested leakage of pulmonary surfactant into the bloodstream caused by damage to the alveolar-capillary barrier in diseased lungs. The ratio of degraded vs. intact SP-D found in the plasma was compared before and after therapeutic surfactant administration. The results indicated that levels of SP-D in plasma and tracheal aspirates together with the ratio of degraded and intact SP-D in the plasma may be useful indicators of the severity of COVID-19 lung disease progression.

Prospective Evaluation of Serial Protein Biomarker Measurements in Sepsis-Induced Acute Respiratory Distress Syndrome

RATIONALE: Protein biomarkers including soluble receptor for advanced glycation end-products (sRAGE), angiopoietin-2 (Ang- 2), and surfactant protein-D (SP-D) have been studied for diagnosis or prognostication in acute respiratory distress syndrome (ARDS). However, prior studies of ARDS biomarkers often included heterogeneous populations, and rarely examined serial measurements. Our aim was to determine the association between serially measured sRAGE, Ang-2, and SP-D levels and ARDS development in patients with sepsis. METHODS: Adult patients admitted to the medical ICU at Grady Memorial Hospital within 72 hours of sepsis diagnosis were enrolled into this prospective observational cohort study within 48 hours of ICU admission. Patients who already had ARDS at the time of screening were excluded. After obtaining informed consent, serial plasma samples were collected on day 1, 2, and 3 of enrollment, and were analyzed for sRAGE, Ang-2, and SP-D levels using ELISA. The patients were followed for up to 28 days for relevant clinical characteristics and outcomes. The primary outcome was ARDS development according to the Berlin Definition. The secondary outcome was mortality. Pulmonary sepsis was defined as the primary infection being pneumonia (including COVID19) or aspiration pneumonia. The biomarker levels and their changes from day 1 to days 2/3 were compared between those who developed ARDS versus those who did not. RESULTS: Among 80 patients with sepsis enrolled between September 1, 2020 and June 22, 2021, 15 patients (18.8%) developed ARDS and 65 patients (81.3%) did not. ARDS patients had higher proportions of pulmonary sepsis (14/15 [93.3%] vs. 30/65 [46.2%], p=0.001) and COVID19 (7/15 [46.7%] vs. 7/65 [10.8%], p=0.003) compared to non-ARDS patients. ARDS patients had higher SP-D levels on days 1 and 2, and had a greater increase in sRAGE levels from day 1 to day 3, compared to non-ARDS patients (Figure 1A- 1B). Within the ARDS group, those who died had higher sRAGE levels on day 1 compared to those who survived (Figure 1C). CONCLUSIONS: In this analysis, ARDS patients had higher SP-D and a greater increase in sRAGE over time compared to non- ARDS patients. Non-survivors of ARDS also had higher sRAGE compared to survivors. Our findings suggest that early serial biomarker measurements may be useful for identifying sepsis patients at risk of developing ARDS and adverse clinical outcomes, and for risk stratifying sepsis patients in ARDS clinical trials focused on early therapeutics and prevention. Larger studies are needed for more detailed analyses and confirmation of these findings.

Correlation of Surfactant Protein-D (SP-D) Serum Levels with ARDS Severity and Mortality in Covid-19 Patients in Indonesia.

OBJECTIVE: The purpose of this research was to investigate the correlation between serum levels of surfactant protein-D (SPD) with acute respiratory distress syndrome (ARDS) severity and mortality in COVID-19. MATERIALS AND METHOD: This was a prospective cohort research study that included 76 patients in the period from July to October 2020. SP-D serum levels were taken upon admission to the hospital, the diagnosis of ARDS and its grade were confirmed according to the WHO criteria, and then patients were observed for 28-day mortality. RESULTS: The mean SP-D serum levels from 76 patients were 39.33 ng/ml (SD±31.884 ng/ml). The statistical analysis showed that there was a significant correlation between SP-D serum levels and the severity of ARDS upon admission to the hospital (P=0.04, Spearman's rank correlation coefficient (rs)=0.26), but the correlation between serum levels of SP-D and mortality was not statistically significant (P=0.89; rs=-0.016). CONCLUSION: SP-D serum levels had a significant but weak correlation with ARDS severity, but were not significant for mortality.

IMPACT OF COVID-19 PANDEMIC ON ADHERENCE TO ORAL ONCOLOGY MEDICATIONS AND QUALITY OF LIFE OF CANCER PATIENTS ON THE SERVICE OF A SPECIALTY PHARMACY

BACKGROUND: In late winter 2019-2021 SARS-CoV-2 became pandemic (CoVID-19 or P). During P, treating patients with chronic disease burden, i.e., cancer (O), with O drugs and achieving high medication adherence was reported to be challenging, e.g., delayed/missed provider visits, prescriptions, lab tests, and P-related financial difficulties and feelings of depression or anxiety. OBJECTIVE: There are two objectives that characterize how the P may be associated with changes in: 1. adherence to specialty pharmacy O medications (O-SP-D) and 2. O patient- reported quality of life (QoL). METHODS: A retrospective, observational, pre-/post- design study of 393 USA O patients' adherence to O-SP-D and their reported QoL was conducted Pre-P (1/1-12/31/2019) and Post-P (3/1-8/31/2020). Patient assessments (PA), including medication therapy management (SPMTM) and the EQ-5D-5L QoL instrument, occurred at start-of-care (SOC) and 7-10 days before refills (F-U). Descriptive statics were used to calculate Pre-P Dimensional means before and after starting O-SP-D with SPMTM and again at F-U Post-P. The means of QoL measures in the Pre-P and Post-P periods were compared for differences, representing improved or diminished QoL associated with P. The mean medication possession ratio (MPR or adherence) was calculated, using the F-U from Pre-P and Post-P. RESULTS: MPR was 0.976 Pre-P and 0.98 during Post-P (0.4% improvement). Differences between QoL means Pre-P vs Post-P were: Mobility -0.31 (31%), Self-Care 0 (no change), Usual Activities -0.02 (2%), Pain/Discomfort -0.09 (9%), Anxiety/Depression -0.3 (30%), Overall Health State +2.37 (2.4%). (Minus (-) is improvement in Dimensions 1-5;Positive (+) is improvement in Overall Health State.) CONCLUSIONS: Post-P MPR improved 0.4% compared to Pre-P, and both were higher than the industry standard of 0.8. On average, for all QoL Dimensions (except Self Care), O patients reported improvements during Post-P compared to Pre-P. Explanations associated with improvements are suggested. Additional studies are suggested.

ASSOCIATION OF COVID-19 WITH ADHERENCE TO ORAL NEUROLOGY MEDICATIONS AND QUALITY OF LIFE OF NEUROLOGY PATIENTS ON THE SERVICE OF A SPECIALTY PHARMACY

BACKGROUND: In late winter 2019-2021 SARS-CoV-2 became pandemic (CoVID-19 or P). During P, treating patients with chronic disease burden, i.e., neurologic (N) conditions with N drugs and achieving high medication adherence was reported to be challenging, e.g., delayed/missed provider visits, prescriptions, lab tests, and P-related financial difficulties and feelings of depression or anxiety. OBJECTIVE: The two objectives are how the P may be associated with changes in: 1. adherence to specialty pharmacy N medications (N-SP-D) and 2. N patient-reported quality of life (QoL). METHODS: A retrospective, observational, pre-/post- design study of 1,652 N patients' adherence to N-SP-D and their reported QoL was conducted Pre-P (1/1-12/31/2019) and Post-P (3/1-8/31/2020). Patient assessments (PA), including medication therapy management (SPMTM) and the EQ-5D-5L QoL instrument, occurred at start-of-care (SOC) and 7-10 days before refills (F-U). Descriptive statics were used to calculate Pre-P Dimensional means before and after starting N-SP-D with SPMTM and again at F-U Post-P. The means of QoL measures in the Pre-P and Post-P periods were compared for differences, representing improved or diminished QoL associated with P. The mean medication possession ratio (MPR or adherence) was calculated, using the F-U from Pre-P and Post-P. RESULTS: MPR was 0.965 Pre-P and 0.975 during Post-P (1% improvement). Differences between QoL means Pre-P vs Post-P were: Mobility +0.53 (53%), Self-Care +0.44 (44%), Usual Activities +0.46 (46%), Pain/Discomfort +0.5 (50%), Anxiety/Depression -0.14 (14%), Overall Health State -7.06 (2.4%). Positive (+) = decrease in Dimensions 1-5;Negative (-) = decrease in Overall Health State. CONCLUSIONS: Post-P MPR increased 1% compared to Pre- P, and both were higher than the industry standard of 0.8. On average, for all QoL Dimensions (except Anxiety/Depression which improved), N patients reported decreased QoL Post-P compared to Pre-P. Explanations associated with improvements are suggested. Additional studies are suggested.

SP-D and CC-16 Pneumoproteins' Kinetics and Their Predictive Role During SARS-CoV-2 Infection.

BACKGROUND: Surfactant protein D (SP-D) and pulmonary club cell protein 16 (CC-16) are called "pneumoproteins" and are involved in host defense against oxidative stress, inflammation, and viral outbreak. This study aimed to determine the predictive value of these pneumoproteins on the incidence of acute respiratory distress syndrome (ARDS) or death in patients with coronavirus disease-2019 (COVID-19). METHODS: This retrospective study included 87 patients admitted to an emergency department. Blood samples were collected on three time points (days 1, 5, and 14 from hospital admission). SP-D and CC-16 serum levels were determined, and univariate and multivariate analyses considering confounding variables (age, body mass index, tobacco use, dyspnea, hypertension, diabetes mellitus, neutrophil-to-lymphocyte ratio) were performed. RESULTS: Based on the multivariate analysis, SP-D level on D1 was positively and slightly correlated with subsequent development of ARDS, independent of body mass index, dyspnea, and diabetes mellitus. CC-16 level on D1 was modestly and positively correlated with fatal outcome. A rise in SP-D between D1 and D5 and D1 and D14 had a strong negative association with incidence of ARDS. These associations were independent of tobacco use and neutrophil-to-lymphocyte ratio. CONCLUSIONS: Overall, our data reveal that increase in SP-D levels is a good prognostic factor for patients with COVID-19, and that initial CC-16 levels correlated with slightly higher risk of death. SP-D and CC-16 may prove useful to predict outcomes in patients with COVID-19.

Host Protein Biomarkers Predicting Severity of Lung Damage due to COVID-19

Background. Biomarkers to predict the severity of lung damage due to COVID-19 are urgently needed to inform management and treatment decisions. Our objective was to investigate the predictive value of host proteins for worsening respiratory failure in one of the by COVID-19 most affected and diverse patient populations in the US. Methods. We performed a prospective single-center cross-sectional study of 34 adult patients admitted to Montefiore Medical Center in the Bronx, New York, for respiratory symptoms due to PCR-confirmed COVID-19. Exclusion criteria were age < 21, history of prior SARS-CoV-2 infection, and/or underlying severe chronic lung diseases requiring home O2 and/or high dose steroids. We stratified and compared patients by whether they developed worsening respiratory failure, necessitating transfer to the intensive care unit (ICU) during their hospital stay. Using a custom Luminex Assay, we measured hospital admission serum concentrations of 8 host proteins, representing respiratory-associated epithelial (RAGE, SP-D, CC16), endothelial (Ang-2, vWF), and immune pathways (S100A12, ICAM-1, VCAM-1). Results. Except for race and WHO COVID-19 scores, demographics, co-morbidities, symptoms, and symptom duration were not statistically significantly different between patients requiring transfer to the ICU (n=15) and non-ICU patients (n=19). Higher log-transformed levels for 5/8 proteins (S100A12, ICAM-1, Ang-2, RAGE, SP-D) showed significant or marginally significant increased cause-specific hazard for ICU transfer (n=15). Estimated cumulative incidence functions further showed a significantly or near significantly increased risk for ICU transfer for patients with above the median values of S100A12 or ICAM-1 (p=0.013), Ang-2 (p=0.056) or RAGE (p=0.077), respectively (Figure 1). Host proteins predicting need for ICU transfer did not correlate strongly with other clinical laboratory markers for COVID-19 severity (CRP, LDH, D-Dimer, Fibrinogen, Ferritin). Comparison of estimated cumulative incidence at 7 days post admission for host protein markers above and below median levels for (A) S10012 (median 96,675 pg/ml);(B) ICAM-1 (median (1,192,277 pg/ml);(C) Ang-2 (median 3463 pg/ml);(D) RAGE (median 6356 pg/ml);and (E) SP-D (median 11,832 pg/ml). Conclusion. These results suggest that host proteins have additional predictive value for the severity of COVID-19-associated lung damage at time of presentation to the hospital.

POSC374 Association of COVID-19 on Adherence to Oral Specialty Pharmacy Oncology Medications and Quality of Life of Multiple Myeloma Patients

Objectives: In winter of 2019-2021 SARS-CoV-2 became pandemic (CoVID-19 or P). During P, treating patients with chronic disease burden, i.e., cancer (O), with multiple myeloma (MM) oncologics and achieving high medication adherence was reported to be challenging, e.g., delayed/missed provider visits, prescriptions, lab tests, and P-related financial difficulties and feelings of depression or anxiety. There are two Objectives: that characterize how the P may be associated with changes in: 1. adherence to specialty pharmacy MM medications (O-SP-D) and 2. MM patient-reported quality of life (QoL). Methods: A retrospective, observational, pre-/post- design study of 199 USA MM patients’ adherence to MM-SP-D and their reported QoL was conducted Pre-P (1/1-12/31/2019) and Post-P (3/1-8/31/2020). Patient assessments(PA), including medication therapy management (SPMTM) and the EQ-5D-5L QoL instrument, occurred at start-of-care (SOC) and 7-10 days before refills (F-U). Descriptive statics were used to calculate Pre-P Dimensional means before and after starting O-SP-D with SPMTM and again at F-U Post-P. The means of QoL measures in the Pre-P and Post-P periods were compared for differences, representing improved or diminished QoL associated with P. The mean medication possession ratio (MPR or adherence) was calculated, using the F-U from Pre-P and Post-P. Results: MPR was 1.0 Pre-P and 0.999 during Post-P (0.1% decrease). Differences between QoL means Pre-P vs Post-P were: Mobility +0.657 (65.7%), Self-Care +0.538 (53.8%), Usual Activities +0.642 (64.2%), Pain/Discomfort+0.884 (88.4%), Anxiety/Depression +0.641 (64.1%), Overall Health State +16.0 (16%). Plus “+” is decreased/worsened in Dimensions 1-5;Positive “+” is improvement in Overall Health State. Conclusions: Post-P MPR decreased 0.1% compared to Pre-P, and both were higher than the industry standard of 0.8. On average MM patients reported decreases all QoL Dimensions Post-P compared to Pre-P. Explanations associated with the decreases are suggested. Additional studies are suggested.

Serum surfactant protein D in COVID-19 is elevated and correlated with disease severity.

BACKGROUND: The serum surfactant protein D (SP-D) level is suggested to be a useful biomarker for acute lung injuries and acute respiratory distress syndrome. Whether the serum SP-D level could identify the severity of coronavirus disease 2019 (COVID-19) in the early stage has not been elucidated. METHODS: We performed an observational study on 39 laboratory-confirmed COVID-19 patients from The Fourth People's Hospital of Yiyang, Hunan, China. Receiver operating characteristic (ROC) curve analysis, correlation analysis, and multivariate logistic regression model analysis were performed. RESULTS: In the acute phase, the serum levels of SP-D were elevated significantly in severe COVID-19 patients than in mild cases (mean value ± standard deviation (SD), 449.7 ± 125.8 vs 245.9 ± 90.0 ng/mL, P<0.001), while the serum levels of SP-D in the recovery period were decreased dramatically than that in the acute phase (mean value ± SD, 129.5 ± 51.7 vs 292.9 ± 130.7 ng/ml, P<0.001), and so were for the stratified patients. The chest CT imaging scores were considerably higher in the severe group compared with those in the mild group (median value, 10.0 vs 9.0, P = 0.011), while markedly lower in the recovery period than those in the acute phase (median value, 2.0 vs 9.0, P<0.001), and so were for the stratified patients. ROC curve analysis revealed that areas under the curve of lymphocyte counts (LYM), C-reaction protein (CRP), erythrocyte sedimentation rate (ESR), interleukin-6 (IL-6), and SP-D for severe COVID-19 were 0.719, 0.833, 0.817, 0.837, and 0.922, respectively. Correlation analysis showed that the SP-D levels were negatively correlated with LYM (r = - 0.320, P = 0.047), while positively correlated with CRP (r = 0.658, P<0.001), IL-6 (r = 0.471, P = 0.002), the duration of nucleic acid of throat swab turning negative (r = 0.668, P<0.001), chest CT imaging score on admission (r = 0.695, P<0.001) and length of stay (r = 0.420, P = 0.008). Multivariate logistic regression model analysis showed that age (P = 0.041, OR = 1.093) and SP-D (P = 0.008, OR = 1.018) were risk factors for severe COVID-19. CONCLUSIONS: Elevated serum SP-D level was a potential biomarker for the severity of COVID-19; this may be useful in identifying patients whose condition worsens at an early stage.

Expression of Surfactant Protein D Distinguishes Severe Pandemic Influenza A(H1N1) from Coronavirus Disease 2019.

The differentiation between influenza and coronavirus disease 2019 (COVID-19) could constitute a diagnostic challenge during the ongoing winter owing to their clinical similitude. Thus, novel biomarkers are required to enable making this distinction. Here, we evaluated whether the surfactant protein D (SP-D), a collectin produced at the alveolar epithelium with known immune properties, was useful to differentiate pandemic influenza A(H1N1) from COVID-19 in critically ill patients. Our results revealed high serum SP-D levels in patients with severe pandemic influenza but not those with COVID-19. This finding was validated in a separate cohort of mechanically ventilated patients with COVID-19 who also showed low plasma SP-D levels. However, plasma SP-D levels did not distinguish seasonal influenza from COVID-19 in mild-to-moderate disease. Finally, we found that high serum SP-D levels were associated with death and renal failure among severe pandemic influenza cases. Thus, our studies have identified SP-D as a unique biomarker expressed during severe pandemic influenza but not COVID-19.

Human Surfactant Protein D Binds Spike Protein and Acts as an Entry Inhibitor of SARS-CoV-2 Pseudotyped Viral Particles.

Human SP-D is a potent innate immune molecule whose presence at pulmonary mucosal surfaces allows its role in immune surveillance against pathogens. Higher levels of serum SP-D have been reported in the patients with severe acute respiratory syndrome coronavirus (SARS-CoV). Studies have suggested the ability of human SP-D to recognise spike glycoprotein of SARS-CoV; its interaction with HCoV-229E strain leads to viral inhibition in human bronchial epithelial (16HBE) cells. Previous studies have reported that a recombinant fragment of human SP-D (rfhSP-D) composed of 8 Gly-X-Y repeats, neck and CRD region, can act against a range of viral pathogens including influenza A Virus and Respiratory Syncytial Virus in vitro, in vivo and ex vivo. In this context, this study was aimed at examining the likely protective role of rfhSP-D against SARS-CoV-2 infection. rfhSP-D showed a dose-responsive binding to S1 spike protein of SARS-CoV-2 and its receptor binding domain. Importantly, rfhSP-D inhibited interaction of S1 protein with the HEK293T cells overexpressing human angiotensin converting enzyme 2 (hACE2). The protective role of rfhSP-D against SARS-CoV-2 infection as an entry inhibitor was further validated by the use of pseudotyped lentiviral particles expressing SARS-CoV-2 S1 protein; ~0.5 RLU fold reduction in viral entry was seen following treatment with rfhSP-D (10 µg/ml). These results highlight the therapeutic potential of rfhSP-D in SARS-CoV-2 infection and merit pre-clinical studies in animal models.

A Recombinant Fragment of Human Surfactant Protein D Binds Spike Protein and Inhibits Infectivity and Replication of SARS-CoV-2 in Clinical Samples.

Coronavirus disease (COVID-19) is an acute infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Human SP-D (surfactant protein D) is known to interact with the spike protein of SARS-CoV, but its immune surveillance against SARS-CoV-2 is not known. The current study aimed to examine the potential of a recombinant fragment of human SP-D (rfhSP-D) as an inhibitor of replication and infection of SARS-CoV-2. The interaction of rfhSP-D with the spike protein of SARS-CoV-2 and human ACE-2 (angiotensin-converting enzyme 2) receptor was predicted via docking analysis. The inhibition of interaction between the spike protein and ACE-2 by rfhSP-D was confirmed using direct and indirect ELISA. The effect of rfhSP-D on replication and infectivity of SARS-CoV-2 from clinical samples was assessed by measuring the expression of RdRp gene of the virus using quantitative PCR. In silico interaction studies indicated that three amino acid residues in the receptor-binding domain of spike protein of SARS-CoV-2 were commonly involved in interacting with rfhSP-D and ACE-2. Studies using clinical samples of SARS-CoV-2-positive cases (asymptomatic, n = 7; symptomatic, n = 8) and negative control samples (n = 15) demonstrated that treatment with 1.67 µM rfhSP-D inhibited viral replication by ∼5.5-fold and was more efficient than remdesivir (100 µM) in Vero cells. An approximately two-fold reduction in viral infectivity was also observed after treatment with 1.67 µM rfhSP-D. These results conclusively demonstrate that the rfhSP-D mediated calcium independent interaction between the receptor-binding domain of the S1 subunit of the SARS-CoV-2 spike protein and human ACE-2, its host cell receptor, and significantly reduced SARS-CoV-2 infection and replication in vitro.

SP-A and SP-D: Dual Functioning Immune Molecules With Antiviral and Immunomodulatory Properties.

Surfactant proteins A (SP-A) and D (SP-D) are soluble innate immune molecules which maintain lung homeostasis through their dual roles as anti-infectious and immunomodulatory agents. SP-A and SP-D bind numerous viruses including influenza A virus, respiratory syncytial virus (RSV) and human immunodeficiency virus (HIV), enhancing their clearance from mucosal points of entry and modulating the inflammatory response. They also have diverse roles in mediating innate and adaptive cell functions and in clearing apoptotic cells, allergens and other noxious particles. Here, we review how the properties of these first line defense molecules modulate inflammatory responses, as well as host-mediated immunopathology in response to viral infections. Since SP-A and SP-D are known to offer protection from viral and other infections, if their levels are decreased in some disease states as they are in severe asthma and chronic obstructive pulmonary disease (COPD), this may confer an increased risk of viral infection and exacerbations of disease. Recombinant molecules of SP-A and SP-D could be useful in both blocking respiratory viral infection while also modulating the immune system to prevent excessive inflammatory responses seen in, for example, RSV or coronavirus disease 2019 (COVID-19). Recombinant SP-A and SP-D could have therapeutic potential in neutralizing both current and future strains of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus as well as modulating the inflammation-mediated pathology associated with COVID-19. A recombinant fragment of human (rfh)SP-D has recently been shown to neutralize SARS-CoV-2. Further work investigating the potential therapeutic role of SP-A and SP-D in COVID-19 and other infectious and inflammatory diseases is indicated.