Določanje koncentracije ustekinumaba z encimsko imunsko metodo v povezavi z vnetnimi parametri pri bolnikih z ulceroznim kolitisom

Bračič, Manca (avtor), Čučnik, Saša (mentor), Drobne, David (komentor)

Ulcerozni kolitis je kronična vnetna črevesna bolezen pri kateri pride do vnetja sluznice gastrointestinalnega trakta. Pri diagnozi bolezni se opravi več preiskav in analiz različnih parametrov, med njimi tudi C-reaktivni protein v krvi in kalprotektin v blatu, ki sta pokazatelja vnetja v telesu. Pri zdravljenju bolezni se vedno več uporabljajo biološka zdravila, kot je ustekinumab. Ustekinumab je človeško monoklonsko protitelo, ki učinkuje z vezavo na podenoto p40 interlevkinov 12 in 23 in prepreči njuno vezavo na receptor. Posledično prepreči aktiviranje celičnega signaliziranja in s tem povezanega vnetnega imunskega odziva. Koncentracije ustekinumaba imajo lahko v določenem tednu zdravljenja napovedno vrednost za odziv bolnika na zdravljenje, kot so ugotovili Hanžel in sodelavci (2020). Namen naloge je bil določiti ustekinumab z dvema različnima analiznima kompletoma ELISA v 165 vzorcih seruma 12 bolnikov z ulceroznim kolitisom in ugotoviti ujemanje med njima ter ugotoviti povezavo med koncentracijami zdravila in vrednostmi vnetnih parametrov (C-reaktivni protein, fekalni kalprotektin) z aktivnostjo bolezni. Analizna kompleta smo med sabo primerjali statistično – Shapiro-Wilkov test za preverjanje normalnosti porazdelitve, Spearmanov test za oceno korelacije, linearno regresijo smo preverili s Passing-Bablokovo regresijo in nato izvedli še Bland-Altmanov test. Znotraj posamezne analize in med analizami smo določili tudi natančnost ter točnost znotraj ene izmed analiz. Pri določenih bolnikih smo v vzorcih določili tudi protitelesa proti ustekinumabu, ki bi lahko motila zdravljenje. Ugotovili smo, da sta analizna kompleta med sabo primerljiva, kar so pokazali rezultati statističnih testov. Ugotovili smo tudi, da pri večini bolnikov ni bilo povezave med koncentracijami C-reaktivnega proteina in zdravila. Večja povezava je bila vidna pri nekaterih bolnikih med vrednostmi fekalnega kalprotektina in ustekinumaba. Primerjali smo tudi koncentracijo ustekinumaba z dosegom biokemijske remisije (<100 mg/kg fekalni kalprotektin) v 16. in 24. tednu zdravljenja – pri obeh analiznih kompletih smo lahko predpostavili, da imajo bolniki, ki imajo v 2. tednu zdravljenja koncentracije ustekinumaba >40 μg/mL, večjo verjetnost za doseg biokemijske remisije v 16. tednu zdravljenja. Od skupno 17 vzorcev 6 bolnikov jih je bilo 6 pozitivnih za prisotnost prostih in 3 pozitivni za prisotnost celokupnih protiteles proti ustekinumabu.

• Jezik: Slovenski jezik
• Ključne besede: ulcerozni kolitis, ustekinumab, ELISA, C-reaktivni protein, fekalni kalprotektin
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: FFA - Fakulteta za farmacijo
• Leto izida: 2024
• PID: 20.500.12556/RUL-158519
• Datum objave v RUL: 14.06.2024

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Determination of ustekinumab concentration by enzyme-linked immunosorbent assay in relation to inflammatory parameters in patients with ulcerative colitis

Izvleček

Ulcerative colitis is a form of chronic inflammatory bowel disease which affects the mucous membrane of the gastrointestinal tract. The diagnosis of the disease is based on a multitude of tests and analyses of different parameters, including C-reactive protein in the blood and calprotectin in the faeces, which are markers of inflammation in the body. For the treatment of the disease, biological drugs, like ustekinumab, are being used more often. Ustekinumab is a human monoclonal antibody that binds to the subunit p40 of the interleukins 12 and 23 and blocks them binding to their receptors. Consequently, it also blocks the activation of cell signalization and the proinflammatory immune response. Hanžel and coworkers (2020) have discovered that concentrations of ustekinumab in a certain week of treatment can have a prognostic value for how the patient will respond to treatment. The purpose of our assignment was to measure ustekinumab with two different ELISA analytical kits in 165 samples of serum of 12 patients with ulcerative colitis and determine their agreement and determine a connection between the concentrations of the drug and inflammation parameters (C-reactive protein, faecal calprotectin) with disease activity. We tested the agreement between kits statistically – Shapiro-Wilk test for the verification of normality of the data, Spearman test for assessment of correlation, we checked the linear regression with the Passing-Bablok regression and then performed the Bland-Altman test. We also calculated the intra- and inter-assay precision for the performed analyses and accuracy for one analysis. In certain patients samples we also checked for the presence of free and total anti-ustekinumab antibodies, which could interfere with treatment. The statistical tests showed good agreement between the kits. We determined there was no connection between C-reactive protein and the drug concentrations in most patients. A better connection was seen between concentrations of faecal calprotectin and ustekinumab. We also compared ustekinumab concentrations with achieving biochemical remission (<100 mg/kg faecal calprotectin) in the 16th and 24th week of treatment – we could assume with both methods that patients, who have a ustekinumab concentration >40 μg/mL in the second week of treatment, have a bigger probability of achieving biochemical remission in the 16th week of treatment. Out of a total of 17 samples from 6 patients, only 6 were positive for the presence of free and 3 were positive for the presence of total anti-ustekinumab antibodies.

• Ključne besede: ulcerative colitis, ustekinumab, ELISA, C-reactive protein, faecal calprotectin