Disseminated intravascular coagulation was induced by injection of thrombin and EACA in rats.In the experimental group the rats, after injection of ~(125)I fibrinogen, received thrombin to produce DIC. The control group received saline instead of thrombin. The autoradiographic result indicated that the radioactive microthrombi were mainly situated in the glomeruli of kidney and pulmonary capillaries. By the use of HE and PTAH stain, however, the microthrombi could be found only in the glomeruli of the kidney but not in pulmonary capillaries. This indicated that autoradiography was more sensitive than histochemical and HE methods for detection of thrombi in microcirculation. It was also discovered that the reticuloendothelial cells of liver and spleen phagocytized a large amount of the radioactive materials. The materials might be radioactive fibrin or its monomer because ~(125)I fibrinogen has been attached to thrombin. In addition to these, the epithelium of convoluded tubules of kidney also absorb radioactive materials which might be peptides A and B.

A Strategy was established for construction of repertoire antibody library with affinity chromatography purifying antigen, antigen immunizing human lymphocytes, RT-PCR and phage display technology. The colorectal carcinoma-associated antigen CA-Hb3 was purified with affinity column and analysed with SDS-PAGE and Western-blot, then applied for immunizing peripheral blood lymphocytes (PBL) in vitro. The PBL were isolated from ten patients with colorectal carcinoma and cultured with interleukin 2 and pokeweed mitogen, lymphoblast-like cells occurred and colonies formed after immunization. Three VH-CHl(?) and five VL-CL(?,?) genes were amplified from their total RNA and mRNA with RT-PCR. Three VH (?) and 8 VL (?,?) genes were reamplified and randomly combined to construct 24 single-chain variable fragments (ScFv) genes through (Gly_(4)Ser)_(3) linker. ScFv genes digested with Sfi I were cloned into fUSE 5 vector and transformed into MC1061 with electroporation. Repertoire antibody library was obtained with 10~(6) tetracycline-resistant colonies, in which the percentage of ScFv inserts was 85 % . This strategy might be used for humanizing mouse-original monoclonal antibody.

According to the immune network hypothesis proposed by Jeme, certain anti-idiotypic antibodies (Ab2) express three dimensional shapes which resemble the structure of natural antigens. Here we propose a study of active immunotherapy by Ab2 for patients with Nasopharyngeal Cancer (NPC) . Two Ab2,designated 2H4 and 5D3,against two Abl (FC_2 and HNL5)that recognize NPC associated antigen were generated. They could substitute NPC antigen to induce humoral and cellular immune response against NPC cells in syngeneic mice. Nineteen patients with NPC at stage IV were chosen for active immunotherapy. They were treated with Alum-precipitated 2H4 or 5D3 accompanying radiotherapy. Nine patients with radiotherapy alone were as control. Both anti-anti-idiotypic antibodies (Ab3) and anti-NPC antibodies (Abl') were increased and human anti-mouse antibodies (HAMA) occurred in nineteen patients of the experimental group; whereas the levels of Abl' did not rise in control group.Serum IL-2, IFN-? and TNF-? level were increased in most patients in experimental group. While in the control group, there was no difference of cytokine level between pretherapy and post- therapy.

OBJECTIVETo investigate the effect of active immunotherapy with anti-idiotypic vaccine in patients with nasopharyngeal carcinoma (NPC).

METHODSAnti-idiotypic antibodies (2H4/5D3) bearing the internal image of the NPC antigen were used in active immunotherapy in NPC patients receiving radiotherapy. Antibodies and cytokine levels in patient sera were determined using ELISA before and after active immunotherapy. IL-2 mRNA expression in the peripheral blood mononuclear cells (PBMC) was measured by in situ hybridization.

RESULTSNineteen patients with NPC at stage IV were treated with alum-precipitated 2H4 or 5D3. Neither hypersensitivity nor adverse side effects were observed. The levels of anti-anti-idiotypic antibodies (Ab3) and anti-NPC antibodies (Ab1') were increased. Human anti-mouse antibodies (HAMA) were seen in 19 patients of the experimental group; the levels of Ab1' did not increase in the control group. Serum IL-2, IFN-gamma and TNF-alpha levels were increased in most patients in the experimental group, while no differences were observed in Ab1' and cytokine levels between pre- and post-therapy in the control group. In addition, IL-2 mRNA expression in PBMCs from NPC patients was closely related to serum IL-2 (r = + 0.8829) levels by in situ hybridization.

CONCLUSIONSAnti-idiotype vaccine is safe for clinical active immunotherapy. Anti-idiotypic vaccine might be able to enhance humoral and/or cellular immunity in NPC patients receiving radiotherapy.

Adult ; Antibodies, Anti-Idiotypic ; immunology ; therapeutic use ; Antibody Specificity ; Cancer Vaccines ; immunology ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunotherapy, Active ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; genetics ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; blood ; drug therapy ; immunology ; RNA, Messenger ; genetics ; metabolism ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; metabolism