Study of Cytokines in Sputum in Pediatric Patients with Cystic Fibrosis, PRE- and Post-Treatment in the Period of Acute Lung Exacerbation

In this study, the clinical characteristics of pediatric patients with cystic fibrosis and the cytokine levels in the sputum before and after lung infection treatment were evaluated to determine lung exacerbation before clinical development and to initiate early treatment. Thus to maintain the patients' existing lung capacity for a longer period of time, to contribute to the ongoing immunotherapy treatment methods. Sputum cytokine study will be performed in 25 children with cystic fibrosis followed in Gaziantep University hospital pediatric allergy-immunology outpatient clinic. Cystic fibrosis diagnosed children with 3-21 age interval by a specialist allergy-immunology physician will be included in this study. As well as, we will include patients with acute lung exacerbation in this study. Informed consent forms will be obtained from the relatives of the participants fort he study. Comparisons of the mediators in the sputum samples taken during acute lung exacerbation and cytokine levels in sputum samples 14-30 days after initiation of treatment will be done. IL-17, IL-8, LTB-4, 8- isoprostane, neutrophil elastase, glutathione, glutathione peroxidase, TLR-2, TLR-4 cytokine levels will be studied by ELİSA method from samples. The level of glutathione in the sputum of the patient with CF was found to be lower during the post-treatment period compared to the level of glutathione in the sputum before the treatment (p:0.002). The level of glutathione peroxidase in sputum in the post-relapse period was significantly lower than the level of glutathione peroxidase in sputum during the attack period (p: 0.058). The TLR-2 level in the sputum before the treatment of patients with increased sputum during the attack period was found to be higher than those in the group without an increase in sputum amount (p: 0.037) IL-8 and TLR-4 levels were found to be higher in sputum (p: 0.041, p: 0.019 respectively) in patients with significant dyspnea during the attack period compared to patients with mild dyspnea in the pre-attack treatment period. LTB4 and glutathione peroxidase levels in the sputum during the attack were found to be higher in the group with complain of wheezing (p: 0.001, p: 0.013 respectively) compared to the group without wheezing complain. LTB4 and IL-8 levels in the sputum (p: 0.011, p: 0.002 respectively) were found to be higher in the sputum during the attack of the patients who had tachypnea at the time of admission compared to the group without tachypnea. The TLR-4 level was found to be lower in the sputum (p: 0.035, p: 0.035 respectively) of the patients who had complaints of weakness and malaise at the time of the attack, compared to the patients who did not have complaints of weakness and malaise. According to the frequency of lung exacerbation, a significant decrease was found in those with IL-8 and TLR-4 levels in sputum during an attack in less than 6 months, once every 6 months, once a year, and for more than 1 year group (p: 0.007, p: 0.032, respectively) According to the frequency of hospitalization, TLR-4 level in the sputum in the sputum after the attack treatment was found to be statistically significant in once every 6 months, once a year, and for more than 1 year groups (p: 0.036). LTB4 and TLR-4 levels in the sputum before the treatment (p: 0.037, p: 0.011, respectively) and TLR-4 levels in the sputum after the treatment (p:0.027) were found to be higher in the patients on whom rales were heard during an attack compared to the patients on whom rales were not heard. There were negative correlation between IL-17 and glutathione in sputum during attack period (p:0.018), negative correlation between LTB4 and glutathione in sputum during attack period (p:0.006), negative correlation between IL-8 and 8-isoprostane in sputum during attack period (p :0.037), a positive correlation between neutrophil elastase and glutathione in sputum during the attack period (p:0.012), and a negative correlation was found between glutathione and TLR-4 in sputum during the attack period (p:0.021) in the patients who have used Kreon. There were a negative correlation between LTB4 and glutathione in the sputum during the attack period of the patients using Dornase alfa (p:0.008), and a negative correlation between IL-8 and 8-isoprostane in the sputum during the attack period (p:0.019), a positive correlation was found between neutrophil elastase and glutathione in sputum during the attack period (p:0.003). According to the microorganism grown in the sputum cultures taken during the attack period, after the attack treatment and 30 days after the end of the treatment, the glutathione level in the sputum of the patients with Staph. Aureus growth was found higher than the patients with P. Aureginosa growth (p: 0.023, p: 0.030, p: 0.026, respectively). There is a positive correlation between IL-8 level in sputum during the attack period and gentamicin MIC value in sputum culture taken at admission (p:0.031), netilmicin MIC value (p:0.025), cefepime MIC value (p:0.034), imipenem MIC value (p:0.016) detected. Positive correlation between netilmicin MIC value in the first sputum culture and IL-8 in sputum during attack (p:0.025),a negative correlation between imipenem MIC value and IL-8 (p:0.016) and TLR-2 (p:0.032) in sputum after attack treatment, a negative correlation was found between netilmicin MIC value and TLR-2 in sputum in the post-relapse treatment period (p:0.035) in the first sputum culture. There was a positive correlation between the imipenem MIC value in the second sputum culture and IL-8 in the sputum during the attack (p: 0.030) and a negative correlation between the 8-isoprostane value (p:0.049), a positive correlation the netilmicin MIC value and IL-8 in the sputum during the attack period (p:0.033, r:0,794) and negative correlation between TLR-2 (p:0.001) in sputum after the attack period treatment in the second sputum culture. The IL-8 and TLR-4 levels in the sputum during the attack period of the patients with P. Aureginosa colonization at the time of admission were found to be higher than the patients without colonization (p: 0.002, p: 0.021, respectively). The LTB4 level in the sputum after the treatment of the patients who had occasional respiratory physiotherapy at the time of admission was found to be higher than the patients who did it regularly (p: 0.036). TLR-4 was positively correlated with CRP value (p: 0.001), white blood cell count (p: 0.015), neutrophil count (p: 0.013) in sputum during the attack period; negative correlation was determined with albumin value (p: 0.009). Negative correlation between lymphocyte count and glutathione in sputum during attack (p: 0.020), positive correlation between hemoglobin and TLR-2 in sputum during attack (p: 0.026), positive correlation between platelet count and IL-8 in sputum during attack (p: 0.045) detected. A negative correlation was found between the number and percentage of eosinophil and IL-8 in the sputum (p: 0.045, p: 0.002 respectively) in the post-attack treatment period. A positive correlation was found between pCO2 and cHCO3 with LTB4 in the sputum during the attack period (p: 0.006, p: 0.004, respectively). A positive correlation was found between vitamin B12 level and TLR-4 in the sputum in the post-attack treatment period (p: 0.023), and between vitamin E level and LTB4 in the sputum in the post-attack treatment period (p: 0.029). In Thorax CT, a negative correlation was found between pathological lung tissue and the volume occupied by the lung, intact lung volume, airway volume, and total lung volume (respectively p: 0.002, p: 0.017, p: 0.008, p: 0.002). A positive correlation was found between pathological lung tissue in thorax CT in the post-attack treatment period and TLR-4 in the sputum during the attack period (p: 0.005), and between IL-17 in the sputum after the attack-treatment period (p: 0.025). In this study, besides the routine examination and imaging methods performed in the follow-up of the patients, the determination of cytokine levels in the sputum can be used to reduce the frequency of attacks in the patients. The treatments provided by considering the cytokine levels and antibiotic resistance in the sputum during the attack period and the post-treatment period will be effective in preventing both lung exacerbation and bacterial colonization. Considering the relationship between hypoxia values and cytokine levels in sputum before the development of PHT with increased mortality in patients with CF, especially LTB4 level in sputum is important and leukotriene antagonists can be used as is the case in asthmatic patients. In patients without thorax CT, cytokine levels in sputum and bronchiectasis can be determined and prevented or the frequency of thorax CT scans can be reduced. By using the data obtained as a result of our study, it will be possible to develop immunotherapy treatments, which are still in progress, in order to prevent the development of attacks in patients with CF. Keywords: Cystic fibrosis, acute lung exacerbation, cytokine level in sputum