ELIZABETH S. RAVECHE
.S. 1972 Seton Hill University
and IL-10 receptors
ranslational Research – new therapies to block malignant B cell growth
The Role of Cytokines (IL-10) in Regulation of B cell Apoptosis
The main focus of the research involves analysis of the immunomodulatory effects of interleukin 10 (IL-10) family of cytokines. IL-10 is an important anti-inflammatory cytokine which also acts as a growth factor for B cells. We have found that in the malignant disease, chronic lymphocytic leukemia (CLL), IL-10 is an important growth factor and prevents cell death, apoptosis. We have used both in vitro analysis of cell cultures, mouse models and samples from patients with CLL to further understand the cell cycle regulation induced by IL-10. We have developed several strategies to decrease IL-10 signaling including: antisense IL-10 (US Patent # 6,184,372 B1 (Feb 6,2001), "Antisense Interleukin 10 and Method of Use"), RNA interferences for IL-10, and soluble IL-10 receptor. We have also genetically engineered a mouse model of CLL, NZB, such that these mice no longer produce IL-10 (NZB IL-10KO see Figure below). This strain does not develop the leukemia and is now on the NCI database for mouse models of cancer. Since we found that IL-10 was an important survival factor in the mouse models of CLL we tested this in human CLL cells and found that antisense IL-10 and soluble IL-10 receptor (which binds IL-10 and prevents it from binding to the IL-10 receptor normally expressed on B cells) both induce cell death in malignant B cells. We searched for other agents that might block IL-10 and found that a soy isoflavone, Genistein, was able to decrease IL-10 and induce apoptosis. The present work continues to identify agents which act by decreasing IL-10 and apoptosis induction in malignant B cells.
Figure above compares data obtained from spleen cells in two types of NZB: A. genetically engineered NZB mice devoid of IL-10 expression. B. Wild-type NZB. In the NZB IL-10KO mice the DNA content (top part A) are normal and the surface marker expression is typical for B cells (indicated by hatched boxes). In contrast, the wild- type NZB which develops leukemia (malignant cells are aneuploid indicated by asterisk) and express abnormal surface markers indicated by arrows.
Figure above is the analysis of the cell cycle of human malignant B cells either untreated, treated with soluble IL- 10 receptor or treated with an irrelevant soluble receptor (IL-22R1). Only the soluble IL-10 receptor induced a G2M block in cell cycle and significant apoptosis indicated by the arrow.
Parker, G., Peng, B, He, M., Gould-Fogerite, S., Chou C., and Raveché, E. In vivo use of antisense IL-10. Antisense Technology in Methods in Enzymology , Vol 314 411-429, 2000.
Parker, G., Fernandes, H., Chong, S., Czarneski, J., Hong, R., Lin, Y., and Raveché, E. Antisense IL-10 abrogrates the inhibitory effect of IL-10 production by transfected tumor cells. Cytokines Cellular and Molecular Therapy, 6:113-119, 2000.
Chong, S.Y., Zhang, M., Lin, Y., Coffman, F., Garcia, Z. , Ponzio, N., and Raveché, E. S. The growth regulatory role of BSAP in NZB malignant B-1 cells. Cancer Immunology Immunotherapy, 50:41-50, 2001.
Zhang, M., Chong, S.Y., and Raveché, E. S. The Role of B-Cell specific activator Protein (BSAP) in the response of malignant B-1 cells to LPS. Experimental Cell Research, 264:233-243, 2001.
Chong, S.Y., Lin, Y., Czarneski, J., Zhang, M., Coffman, F., Kashanchi, F., and Raveché , E. S. Cell Cycle Effects of IL-10 on Malignant B-1 Cells. Genes and Immunity, 2:239-247, 2001.
Fernandes, H, Koneru, B, Fernandes, N, Hameed, M., Cohen, M., Raveche, E., and Cohen, S. Investigation of promoter polymorphisms in the TNF alpha and IL-10 genes in the liver of transplant patients. Transplantation, 83:1886-1891, 2002.
Czarneski, J., Lin, Y., Chong,S., McCarthy, B., Parker, G., Fernandes, H., DeCotiis, C., Huppi, K., Marti, G, Mansour, A., and Raveche, E. Studies in NZB IL-10 knockout mice of the requirement of IL-10 for progression of B Cell lymphoma. Leukemia, 18:597-60, 2004.
McCarthy, B., Mansour, A., Lin, Y-C., Kotenko, S., and Raveche, E. RNA interference of IL-10 in leukemic B-1 cells., Cancer Immunity, (in Press), 2004.