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ANTITUMOR EFFECTS OF MONOCLONAL ANTIBODY FAB’FRAGMENT—CONTAINING immunoconjugates

Chinese Medical Sciences Journal 2002年第1期17卷 1-6页

作者： LiuXiaoyun ZhenYongsu InstituteofMedicinalBiotechnology CAMS＆PUMGBeijing100050

*** monoclonal antibody (mAb) Fab′ fragment to develop mAb immunoconjugates for cancer. ***′ fragment of mAb 3A5 was prepared by digestion of the antibody with pepsin and then reduced by dithiothreitol (DTT),while Fab′ fragment of mAb 3D6 was obtained by digestion of the antibody with ficin and subsequently reduced by β *** conjugation between Fab′ fragment and pingyangmycin (PYM),an antitumor antibiotic,was mediated by dextran T *** of Fab′ PYM conjugates with cancer cells was determined by ELISA,and the cytotoxicity of those conjugates to cancer cells was determined by clonogenic *** effects of the Fab′ PYM conjugates were evaluated by subcutaneously transplanted tumors in mice. *** molecular weight of Fab′ fragment was approximately 53 kD,while the average molecular weight of Fab′ PYM conjugate was 170 *** Fab′ PYM conjugates showed immunoreactivity with antigen relevant cancer cells and selective cytotoxicity against target *** intravenously,Fab′ PYM conjugates were more effective against the growth of tumors in mice than free PYM and PYM conjugated with intact mAb. ***′ PYM conjugate may be capable of targeting cancer cells and effectively inhibiting tumor growth,suggesting its therapeutic potential in cancer treatment.

关键词： monoclonal antibody therapeutics Fab′ fragment immunoconjugates antineopla stic agent

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Construction of humanized carcinoembryonic antigen specific single chain variable fragment and mitomycin conjugate

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World Journal of Gastroenterology 2007年第43期13卷 5765-5770页

作者： De-Jie Chen Zui Tan Feng Chen Tian Du Department ofGeneral Surgery Zhongnan Hospital of Wuhan University Wuhan430071 Hubei Province China

AIM： To construct a new target-oriented conjugate of humanized carcinoembryonic antigen （CEA） specific single chain variable fragment （scFv） and mitomycin （MMC） against colorectal cancer, and to investigate its influence on the growth and apoptosis of colorectal cancer cells. METHODS： The primer was designed according to the gene sequence described in reference 16, which respectively contains restriction enzyme cleavage sites BamH I and EcoR I in its upstream and downstream. PCR was performed with the plasmid as template containing genes of humanized anti-CEA scFv. The product was digested by BamH I and EcoR I, and connected to an expression vector which also has the restriction enzyme cleavage sites BamH I and EcoR. Expression of the reaction was induced by isopropy-β -D-thiogalactoside （IPTG）. Then the expression product was covalently coupled with MMC by dextran T-40. The immunoreactivity of the conjugate against colorectal cancer cells as well as CEA was measured by enzyme linked immunosorbent assay （ELISA）. The inhibiting ratio of conjugate on the growth of colorectal cancer cells was also measured by ELISA. The effect of conjugate on the apoptosis of colorectal cancer cells was determined by flow cytometry （FCM）. RESULTS： Restriction endonuclease cleavage and gene sequencing confirmed that the expression vector was successfully constructed. Sodium dodecyl sulfate polyacrylamide gel electropheresis （SDS-PAGE） confirmed that this vector correctly expressed the fusion protein. ELISA confirmed that the conjugate had quite a strong immunoreactivity against colorectal cancer cells and CEA. The conjugate had inhibitory effects on colorectal cancer cells in a concentration-dependent manner and could induce apoptosis of colorectal cancer cells in a concentration-dependent ***： The CEA-scFv-MMC conjugate can be successfully constructed and is able to inhibit the growth and induce apoptosis of colorectal cancer cells.

关键词： Carcinoembryonic antigen Single chainvariable fragment Mitomycin immunoconjugates Colorectal cancer

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Immunotherapy in the treatment of lymphoma

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World Journal of Stem Cells 2021年第6期13卷 503-520页

作者： Lazar S Popovic Gorana Matovina-Brko Maja Popovic Milica Popovic Ana Cvetanovic Ivan Nikolic Biljana Kukic Dragana Petrovic Department for Medical Oncology Oncology Institute of VojvodinaUniversity of Novi SadNovi Sad 21000Serbia Department for Medical Oncology Oncology Institute of VojvodinaNovi Sad 21000Serbia Department for Nephrology and Clinical Immunology Clinical Center of VojvodinaUniversity of Novi SadNovi Sad 21000Serbia Department for Medical Oncology Clinical Center of NisUniversity of NisNis 18000Serbia

Relapsed or refractory non-Hodgkin’s lymphomas,especially diffuse large B-cell lymphoma as well as relapsed or refractory Hodgkin lymphomas are hard-to-treat *** who do not respond to initial therapy or experience relapse are treated with salvage regimens,and if eligible for aggressive therapy,treatment is continued with high-dose chemotherapy and autologous stem cell *** therapy options can cure substantial numbers of patients,however for some it is still an uncurable *** new drugs and cell therapies are being investigated for the treatment of relapsed or refractory *** types of immunotherapy options have shown promising results,and some have already become the standard of ***,we review immunotherapy options for the treatment of lymphoma and discuss the results,positions,practical aspects,and future directions of different drugs and cellular therapies for the treatment of this disease.

关键词： Immunotherapy Receptors Chimeric antigen Antibodies Monoclonal immunoconjugates Hodgkin disease Lymphoma Large B-cell Diffuse

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SPECIFIC UPTAKE OF MONOCLONAL ANTIBODY-CONJUGATED METHOTREXATE BY HUMAN LYMPHOCYTIC LEUKEMIC B CELLS

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Chinese Journal of Cancer Research 1998年第2期10卷 8-17页

作者：朱祯平杨纯正

Objective: To analysis the uptake of free MTX and MTX conjugated to tumor specific monoclonal antibody by target and nontarget cells. Methods: The folate antagonist methotrexate (MTX) was conjugated to two monoclonal antibodies (Mab) directed against human chronic lymphocytic leukemia (CLL), Dal B01 and Dal B02, by an active ester method. Both conjugates were more cytotoxic toward the target tumor cell line D101 than to the nontarget cell line MOLT3, and Dal B02MTX conjugate was more inhibitory to D101 cells than free MTX in a 6 h pulse exposure assay. Results: Drug uptake studies revealed that D101 cells took up much more Dal B01 and Dal B02conjugated MTX than free MTX. The amounts of drug taken up by D101 cells incubated with Dal B01 and Dal B02conjugated MTX were always 3 to 5fold higher than that taken up by MOLT3 cells, although the latter took up more drug when incubated with free MTX. Furthermore, tumor cells incubated with Dal B01 or Dal B02conjugated MTX retained much larger amounts of drug for a prolonged period of time than those incubated with free MTX. Conclusion: The enhanced specific cytotoxicity of Dal B01 and Dal B02MTX conjugates toward target tumor cells is therefore likely due to (I) delivery of larger amounts of MTX to target cells when the drug is conjugated to Mab; (ii) longer retention of Mabconjugated MTX by target cells; and (iii) slow, prolonged release of MTX from the surfacebound or endocytosed conjugates, rendering them into a sustained release dosage form.

关键词： Monoclonal antibody Methotrexate immunoconjugates Drug uptake Specific Cytotoxicity Chronic lymphocytic leukemia.

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CTLA-4 gene A/G polymorphism associated with diabetes mellitus in Han Chinese

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Chinese Medical Journal 2002年第8期115卷 1248-1250,157-158页

作者：马芸汤旭磊常薇高林李茂欣严文伟兰州医学院附属一院内分泌科兰州730000 四川大学第一大学医院普通外科成都610041

OBJECTIVE: To investigate the association of cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) gene A/G polymorphism with susceptibility to diabetes mellitus in Han Chinese. METHODS: An A/G transition at position 49 of exon 1 was analyzed in 31 patients with type 1 diabetes, 31 patients with type 2 diabetes, and 36 controls were analyzed by polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: A highly significant increase in the frequency of the G allele was seen in patients with type 1 diabetes compared with controls (66.1 % vs. 34.7%, respectively; P

关键词： immunoconjugates Polymorphism, Genetic Antigens, Differentiation China Diabetes Mellitus Humans

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