Avtoprotitelesa proti interlevkinu 6 in serumskemu amiloidu A pri bolnikih s koronavirusno boleznijo 2019

Gričar, Eva (avtor), Čučnik, Saša (mentor)

Pri hujšem poteku koronavirusne bolezni 2019 so nivoji vnetnih dejavnikov, kot sta interlevkin 6 (IL-6) in serumski amiloid A (SAA), izrazito povišani. Vlogo pri regulaciji teh dejavnikov bi lahko igrala tudi proti njim usmerjena avtoprotitelesa. Pri raziskovalnem delu znotraj magistrske naloge smo zato želeli določili vrednosti avtoprotiteles proti IL-6 in proti SAA v serumskih vzorcih zdravih krvodajalcev in bolnikov s koronavirusno boleznijo 2019 ter te vrednosti preučiti. Za določanje protiteles proti IL-6 smo postavili in ovrednotili novo hišno metodo ELISA. Določili smo potrebne parametre za ustrezno natančnost metode, specifičnost metode pa smo dokazali z inhibicijo v tekoči in v trdni fazi. V skupini krvodajalcev smo opazili, da spol in starost posameznika ne vplivata na vrednosti protiteles proti IL-6 in proti SAA. Glede na izmerjene nivoje protiteles proti SAA ni bilo razlik med skupinama, bolniki pa so imeli v primerjavi s krvodajalci signifikantno višje vrednosti protiteles proti IL 6 (p = 0,0018). Višja raven protiteles proti IL 6 pri bolnikih nakazuje na to, da ta protitelesa morda tvorijo komplekse z IL-6, ki bi lahko podaljšala čas zadrževanja IL-6 v krvnem obtoku. Preverjali smo tudi prisotnost korelacij med preiskovanimi dejavniki in opazili signifikantno korelacijo med protitelesi proti IL-6 in protitelesi proti SAA v obeh skupinah. V skupini krvodajalcev smo opazili tudi korelacijo med IL 6 in protitelesi proti SAA. Da bi ugotovili, kako se vrednosti preiskovanih protiteles spreminjajo tekom bolezni, smo nekaj izbranih bolnikov spremljali longitudinalno. Vrednosti preiskovanih protiteles so pri večjem deležu pregledanih bolnikov v času hospitalizacije upadle. Pri protitelesih proti SAA smo preverili tudi nevtralizacijsko sposobnost s funkcijskimi testi na celicah. Uporabljeni serumi so SAA nespecifično inhibirali, pri izolirani frakciji protiteles IgG pa nevtralizacijske sposobnosti nismo opazili.

• Jezik: Slovenski jezik
• Ključne besede: covid-19, imunski odziv, ELISA, protitelesa proti IL-6, protitelesa proti SAA
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2022
• PID: 20.500.12556/RUL-137525
• Datum objave v RUL: 21.06.2022

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Autoantibodies against interleukin 6 and serum amyloid A in patients with COVID-19

Izvleček

Levels of inflammatory factors such as interleukin 6 (IL-6) and serum amyloid A (SAA) are distinctly elevated in severe cases of coronavirus disease 2019. A role in regulation of such factors could be attributed to autoantibodies against them. The aim of our research was to determine the values of autoantibodies against IL-6 and against SAA in sera samples of healthy blood donors and coronavirus disease 2019 patients and to analyze these values. To determine the values of antibodies against IL-6 we set up and validated a new in-house ELISA. We determined the needed parameters for the adequate precision of the method and verified the specificity by fluid phase and solid phase inhibition. In the healthy blood donors’ group, we noticed that gender and age of an individual don’t influence the value of the antibodies against IL-6 and SAA. There was no significant difference in the levels of antibodies against SAA between the two groups, but the patients had significantly higher values of antibodies against IL-6 compared to blood donors (p = 0,0018). The higher levels of those antibodies among patients suggest that the antibodies might form complexes with IL-6, that could prolong their half-life in the circulation. We also checked for correlations among the examined factors and observed the significant correlation between antibodies against IL-6 and against SAA in both groups. We also noticed a significant correlation between IL-6 and antibodies against SAA. To see how the values of the examined antibodies change during the disease we chose the samples of some of the patients for longitudinal observation. The levels of the examined antibodies mostly decreased during the time of hospitalization. We also checked the neutralizing ability of the antibodies against SAA with functional tests on cells. The used sera have non-specifically inhibited SAA, but there was no neutralizing ability to be observed with isolated IgG fractions.

• Ključne besede: COVID-19, immune response, ELISA, antibodies against IL-6, antibodies against SAA