Hodgkin lymphoma (HL) is a biologically diverse group of lymphoid tumors, which accounts for 1% of all de novo neoplasms in the world’s population. It is divided into two main groups: the more common classic Hodgkin lymphoma (cHL) and the less common nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL). cHL is further divided into four subtypes, which differ in morphology and the contents of tumor microenvironment. Macrophages are one of the components of tumor microenvironment known to contribute to creating an immunosuppressive microenvironment, which inhibits the activity of cells expressing granzyme B against tumor cells, even when tumor cells are infected with Epstein–Barr virus (EBV). Our research aimed to explore the association between the specific contents of tumor microenvironment and the genetic anomalies in tumor cells. The presence and the relative percentage of cytotoxic T lymphocytes and macrophages was detected by immunohistochemical staining of the antigens specific for certain cell populations. Fluorescent in situ hybridization was used to detect anomalies in the genome of tumor cells and in situ hybridization was used to detect the presence of EBV. Our results show an association between the number of CD163+ macrophages and the number of TP53 copies or BCL6 gene translocation. Patients who had a higher number of CD163+ macrophages infiltrating tumor tissue and three or higher number of copies of TP53 showed poorer survival. We conclude that the presence of macrophages may contribute to genetic instability in cHL, which drives the progression of cHL and decreases survival of the patients.

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SažetakImunohistokemijsko bojenje jest metoda detekcije prisutnosti i lokalizacije specifičnih antigena u stanici ili tkivu. Primjenjuje osnovni princip imunosne reakcije kod kojeg se specifično antitijelo veže na određeni antigen koji prepoznaje. Razvoj imunohistokemijskih metoda započeo je 1930-ih godina, od kada se, sve do danas, primjenjuju u različitim oblicima za mnoga znanstvena istraživanja i u rutinskim dijagnostičkim postupcima.U ovom radu opisan je razvoj metoda imunohistokemijskog bojenja te je prikazana optimizacija protokola za detekciju antigena bACH2 u ljudskom sekundarnom limfnom tkivu. Raspravljeni su pojedini koraci metode poput pripreme prereza tkiva koje se analizira, uvjeti demaskiranja antigena i inkubacije specifičnim antitijelom te je predložen optimalan protokol bojenja za naveden antigen primjenjiv u rutinskom hematopatološkom laboratoriju.

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