engleski

Expression of perforin in fallopian tuba ectopic pregnancy

Leukocytes (CD45+) endowed intrauterine decidua of early normal and ectopic pregnancy. T lymphocytes predominate in tubal implantation site and distal tubal mucosa of ectopic pregnancy. Contrary to T lymphocytes, CD56+ NK cells predominate in intrauterine decidua of normal and ectopic pregnancy. Both, T and NK cells of normal human pregnancy decidua produce and harbor substatial numbers of cytolytic molecule-perforin, indicating their high cytolytic potential. Interleukin-15 (IL-15) is the most important factor that upregulates perforin expression during normal pregnancy. Although the role of perforin at the materno-fetal interface is not yet completely understood, this protein could be involved in the control of trophoblast growth and early placentation. The aim of our study was to examine the presence and distribution of perforin and IL-15 in fallopian tuba ectopic pregnancy. Tubal mucosa at the implantation site and away from the implantation site, as well as intrauterine decidua was obtained from the women whose ectopic pregnancy had been terminated by surgical removal. Perforin and IL-15 cytokine expression was analyzed by immunohistology using HRP kit (Dako) in each of the mucosal site and compared with perforin expression in normal human pregnancy decidua. Mucosal mononuclear cells, obtained by enzymatic digestion and Ficoll density gradient centrifugation were double labeled by immunofluorescency. Intracellular antigen perforin was stained by mouse anti-human perforin IgG2b monoclonal antibody using cell permeabilisation method. Cell surface antigens (CD3, CD8, CD56) were labeled simultaneously. Samples were analyzed by flow cytometry within lymphocyte gate. Many perforin positive (P+) and IL-15+ cells were found in situ in intrauterine decidua of ectopic and normal human pregnancy, while P+ and IL-15+ cells were rare in tubal mucosa of ectopic pregnancy. Flow cytometry showed that 50% of lymphocytes from intrauterine decidua are P+, while only about 5% of P+ cells were found within lymphocytes of tubal mucosa at and away from the implantation site. In intrauterine decidua of ectopic pregnancy perforin was mostly distributed in CD56+ NK cells, while in both sites of tubal mucosa perforin was distributed up to 60% in CD56+ NK cells and about 30-40% in CD3+ lymphocytes. The presence and distribution of P and IL-15 in intrauterine decidua of a women with ectopic pregnancy resembles to those of normal pregnancy. Low percentage of P+ and IL-15+ cells at and away from the implantation site in the fallopian tuba could be responsible for insufficient control of trophoblast growth in ectopic pregnancy. Supported by Ministry of Science and Technology, Republic of Croatia (Grant No.0062029).

perforin; IL-15; pregnancy; decidua

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