Sestre Milosrdnice University hospital and Institute of Clinical Medical Research
Abstract
The aim of the study was to determine the possible bone marrow involvement in patients with early stages of classic Hodgkin\u27s disease mixed cellularity variant diagnosed by lymph node biopsy at initial presentation not responding to radiotherapy alone. The study cohort consisted of 20 patients (18 displaying B-cell genotype and two T-cell genotype) with stages I-II Hodgkin\u27s disease according to Ann Arbor classification treated with radiotherapy alone, seven of them not responding to therapy. Southern blot hybridization using a specific EBV Bam H1W fragment probe showed the presence of EBV genomes in two patients. All 20 patients underwent iliac crest trephine biopsy and a panel of antibodies including CD45, CD20, CD4, CD8, CD45RO, CD56, CD30, ALK-1, CD-15, EMA, CD61, and CD68 were performed. A statistically significant predominance of CD45, CD45RO and CD4 positive stromal cells was found in seven patients that failed to respond to therapy (c2-test: p=0.021, p=0.019 and p=0.015, respectively). The predominance of CD4 positive cells in the bone marrow stroma might be suggestive of involvement by Hodgkin\u27s disease in the early stage (I-II) patients (indicating upstaging) who fail to show remission on radiotherapy alone, and could explain the abnormal cytokine production, which may contribute to diminished T-cell immunity and inefficient antitumor responses despite a vast majority of infiltrating reactive immune cells.Cilj rada bio je utvrditi moguću zahvaćenost koštane srži u bolesnika s ranim stadijima klasične Hodgkinove bolesti, varijanta miješane celularnosti, dijagnosticirane biopsijom limfnog čvora, koji pri prvom dolasku nisu odgovorili na samu radioterapiju. Bolesnička skupina sastojala se je od 20 bolesnika (18 s B-staničnim genotipom i dvoje s T-staničnim genotipom) u I.-II. stadiju prema klasifikaciji iz Ann Arbora, koji su bili liječeni samo radioterapijom. Sedmoro bolesnika nije odgovaralo na ovu vrst liječenja. Southern blot hibridizacija uz primjenu specifične fragmentne sonde EBV Bam H1W pokazala je prisutnost EBV genoma u dvoje bolesnika. Svih 20 bolesnika podvrgnuto je trepanacijskoj biopsiji ilijačnog grebena i provedeno je ispitivanje panelom protutijela uključujući CD45, CD20, CD4, CD8, CD45RO, CD56, CD30, ALK-1, CD-15, EMA, CD61 i CD68. U sedmoro bolesnika koji nisu odgovarali na liječenje utvrđena je statistički značajna prevlast stromalnih stanica pozitivnih na CD45, CD45RO i CD4 (c2-test: p=0,021, p=0,019 odnosno p=0,015). Prevlast stanica pozitivnih na CD4 u stromi koštane srži moglo bi ukazivati na zahvaćenost koštane srži Hodgkinovom bolešću u bolesnika s ranim stadijem (I.-II.) (tj. porast stadija) u kojih ne dolazi do remisije bolesti nakon same radioterapije, te bi moglo objasniti nenormalnu proizvodnju citokina, što možda doprinosi smanjenoj T-imunosti i nedostatnom protutumorskom odgovoru usprkos goleme većine infiltriranih reaktivnih imunih stanica
