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291.
292.
There is often an interest in knowing, for a given ligand concentration, how many protein molecules have one, two, three, etc. ligands bound in a specific manner. This is a question that cannot be addressed using conventional ensemble techniques. Here, a mathematical method is presented for separating specific from nonspecific binding in nonensemble studies. The method provides a way to determine the distribution of specific binding stoichiometries at any ligand concentration when using nonensemble (e.g., single-molecule) methods. The applicability of the method is demonstrated for ADP binding to creatine kinase using mass spectroscopy data. A major advantage of our method, which can be applied to any protein-ligand system, is that no previous information regarding the mechanism of ligand interaction is required. 相似文献
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294.
A. Eisenthal Yechiel Goldman Yehuda Skornick Anna Gelfand Diana Buyaner Issac Kaver Alon Yellin Henry Yehoshua Beatriz Lifschitz-Mercer Amnon Gonnene M. Shinitzky 《Cancer immunology, immunotherapy : CII》1998,46(6):304-310
Hydrostatic pressure (P) combined with membrane protein crosslinking (CL) by adenosine dialdehyde (AdA) can render tumor
cells immunogenic. We have recently shown that PCL treatment of murine tumor cells augmented the presentation of MHC-restricted
tumor-associated antigens and enhanced cell-mediated immunity. In cancer patients inoculated with autologous PCL-modified
tumor cells, a significant delayed-type hypersensitivity response was elicited. Since the balance between cell-mediated immunity
and humoral immunity is reciprocally controlled by immunoregulatory cytokines, we have examined the proliferative response
and cytokine secretion pattern in cultures of human peripheral blood mononuclear cells (PBMC) stimulated by autologous PCL-modified
and unmodified tumor cells. These tumor cells were obtained from freshly resected tumor tissue of 16 patients with colon (8),
lung (4) and renal (4) carcinomas. The results demonstrated that PCL-modified tumor cells promoted an increase in PBMC proliferation
in 5 out of 8 (63%), 1 out of 4 (25%) and 4 out of 4 (100%) colon, lung and renal cell carcinomas. Fourteen of the above cultures
were also analyzed for the secretion of interleukin-10 and interferon-γ. Overall, a substantial decrease in IL-10 secretion
was detected in 9 out of 14 (64%) cultures while a reciprocal increase in interferon-γ secretion was noted in 8 out of 14
(57%) cultures. Our results confirmed that PCL-modified human tumor cells of different etiologies can modulate the pattern
of cytokines released from stimulated autologous lymphocytes. Such a procedure could prove valuable in the production of autologous
tumor vaccines.
Received: 8 January 1998 / Accepted: 9 April 1998 相似文献