Preparation and functional analysis of tumor-infiltrating stroma cells using bone marrow chimera mice.
Tumor-infiltrating stroma cells (TISC) as well as tumors themselves are thought to be involved in tumor-related immunosuppression, which is one of the critical mechanisms of tumor escape from immune surveillance. However, preparation of TISC is difficult because of the small proportion of TISC in established tumors. Thus, the cells thought to be involved in tumor-related immunosuppression are generally prepared from spleens or draining lymph nodes in tumor-bearing mice. In this study, we developed a method for directly preparing TISC from established tumors in order to analyze their function. Using green fluorescent protein (GFP) transgenic (Tg) mice and C57BL/6 mice transplanted with bone marrow (BM) cells of GFPTg mice, we detected three subpopulations of TISC: one is compatible with immature myeloid cells (ImC) derived from BM and the two other subpopulations, CD11b(+) cells and CD11b(-) cells, do not originate from BM. The TISC including these subpopulations but not each subpopulation independently after culturing with tumors in the presence of GM-CSF could suppress T cell proliferation induced by anti-CD3. In our system, tumors did not inhibit T cell responses directly, but unknown factors from tumors affected immunosuppression by TISC.
Ishigaki
Hirohito
H
Department of Gastroenterology and Hematology, Shiga University of Medical Science, Ohtsu, Shiga, Japan.
Yamamoto
Yoshiko
Y
Ishida
Hideaki
H
Kajino
Kiichi
K
Itoh
Yasushi
Y
Fujiyama
Yoshihide
Y
Ogasawara
Kazumasa
K
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology*; D016026:Bone Marrow Transplantation / Q000276:immunology; D039481:CD11b Antigen / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D007108:Immune Tolerance; D007157:Immunologic Surveillance; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009368:Neoplasm Transplantation; D017154:Stromal Cells / Q000166:cytology / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*; D018800:Thy-1 Antigens / Q000276:immunology; D013945:Thymoma / Q000276:immunology*; D016212:Transforming Growth Factor beta / Q000276:immunology / Q000378:metabolism; D018183:Transplantation Chimera; D019139:Tumor Escape
D039481:CD11b Antigen; D018800:Thy-1 Antigens; D016212:Transforming Growth Factor beta; D016753:Interleukin-10