title: Fine-tuning the EBV+ hu-PBL-SCID xenogeneic chimera model using in vivo superinfection.
abstract: Our purpose was to establish a reproducible xenogeneic chimera model to observe tumors similar to the well-known human posttransplant lymphoproliferative disease (LPD). First we followed the original protocol injecting Epstein-Barr virus positive (EBV+) human peripheral blood lymphocytes (PBL) intraperitoneally into immunodeficient (SCID) mice. Human cells showed T cell phenotype in majority one week after the transfer, whereas one month later a shift towards B cell phenotype was evident according to immunohistochemical and flow cytometric analysis. At this stage the intraperitoneal mass of cells suggested a biologically malignant behaviour infiltrating the liver and the spleen of the host animal. Immunohistochemistry indicated proliferating human lymphatic cells expressing EBV associated proteins and characteristic patterns of invasion within the affected organs. Eventually LPD was lethal to the host animals in 46-67 days. However, the microscopic appearance of experimental LPD was different from the human haemopoietic malignancies: the basic structures of lymphatic organs were preserved and the human T and B cells repopulated the normally T and B dependent areas in mice. The phenotypes of the proliferating cells were human and characteristic for the mature T- and B-lymphocytes. No dominant clone developed during in vitroculturing of the biologically invasive mass of cells removed from the tumor-bearing mice. The results of microscopical, immunological, and flow cytometrical analysis suggested a mature but uncontrolled proliferation of human lymphocytes in SCID mice. The original method for the induction of post-transplant LPD in SCID mice was modified in our further experiments to standardise the experimental technique increasing the efficiency of B cell proliferation and the reducing the number of unspecific factors. Subsequent in vivo EBV superinfection was carried out after the intraperitoneal transfer of a reduced quantity of human PBL from different donors. The same disease developed in our modified chimera model as by the use of original protocol except for some valuable differences. All hosts developed LPD regardless the significantly reduced amount of transplanted PBL and it was lethal in a shorter period of time (41-43 days) compared to the original model. The decreased quantity of transplanted human lymphatic cells was formerly insufficient using the original protocol. Therefore this modified and standardised protocol can lead to a more predictable and reproducible model allowing us to examine fine details of posttransplant lympho-proliferative disease.
authors.affiliation: University of Pécs, Faculty of Medicine, Department of Immunology and Biotechnology Szigeti u. 12., Pécs, H-7643, Hungary.
authors.forename: K
authors.initials: K
authors.lastname: Kvell
authors.forename: P
authors.initials: P
authors.lastname: Balogh
authors.forename: P
authors.initials: P
authors.lastname: Németh
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D020031:Epstein-Barr Virus Infections / Q000150:complications*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D066298:In Vitro Techniques; D017710:Lymphocyte Transfusion; D008232:Lymphoproliferative Disorders / Q000473:pathology; D051379:Mice; D016513:Mice, SCID; D023421:Models, Animal; D015163:Superinfection; D018183:Transplantation Chimera; D014183:Transplantation, Heterologous