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A study of Hodgkin/Reed-Sternberg cells using single cell polymerase chain reaction

Chinese Medical Journal 2000年第1期113卷 65-69页

作者：邓飞廖黎明吕广能李甘地杨光华 China 563003 China 563003 China 610041 China 610041

OBJECTIVE: To investigate the characteristics of Hodgkin/Reed-Stemberg (H/R-S) cells found in patients with various types of Hodgkin's disease (HD). METHODS: H/R-S cells were micropicked from frozen sections of tissues affected by HD. The DNA from these cells was amplified by polymerase chain reaction (PCR) using immunoglobulin heavy chain gene FR III a/JH primers and light chain gene family-specific primers. RESULTS: A total of 52/135 (35.8%) isolated cells showed the specific products in the reactions. IgH and V kappa 4 rearrangements were repeatedly found in many cells from a lymphocyte predominance type sample; repeated V kappa 4 and individual IgH/V kappa 2,4 rearrangements and individual IgH, V lambda 3/ V kappa 4 rearrangements were found in two different cases of the nodular sclerosis type; repeated IgH/ V lambda 3 and individual V lambda 2,4 rearrangements, repeated V kappa 2,4 rearrangements, repeated V kappa 4 and individual IgH/ V kappa 3 rearrangements, repeated IgH and individual V kappa 3/ V lambda 4 rearrangements were detected in 3 cases of the mixed cellularity type. Repeated and individual IgH rearrangements were found in other 2 cases. CONCLUSION: The H/R-S cells isolated from the lymphocyte predominance subtypes of HD have IgH and V lambda 4 gene rearrangements. This suggests that the lymphocyte predominance type is a proliferation of neoplastic B cells. The cells isolated from the mixed cellularity and nodular sclerosis types derive from B lineage cells at various stages of differentiation because of the presence of their IgH, kappa and/or lambda gene rearrangements. To our knowledge, this is the first time that the lambda gene rearrangement was detected in H/R-S cells.

关键词： Gene Rearrangement genes, immunoglobulin Hodgkin Disease Humans immunoglobulin Heavy Chains immunoglobulin Variable Region immunoglobulin kappa-Chains immunoglobulin lambda-Chains Polymerase Chain Reaction Reed-Sternberg Cells

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Construction and selection of the natural immune Fab antibody phage display library from patients with colorectal cancer

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World Journal of Gastroenterology 2001年第6期7卷 811-815页

作者： Bao-Ping Wu~1 Bing Xiao~1 Tian-Mo Wan~1 Ya-Li Zhang~1 Zhen-Shu Zhang~1 Dian-Yuan Zhou~1 Zhuo-Sheng Lai~1 Chun-Fang Gao~2 1 Institute for Digestive Diseases,Nanfang Hospital,Guangzhou 510515,Guangdong Province,China2 Surgical Department of Colon and Rectum,150 Central Hospital,Luoyang 471031,Henan Province,China Institute for Digestive Medicine Nanfang Hospital Guangzhou 510515 Guangdong Province China. bpwu@***

AIM: To construct the natural immune Fab antibody phage display libraries of colorectal cancer and to select antibodies related with colorectal cancer. METHODS: Extract total RNA from tissue of local cancer metastasis lymph nodes of patients with colorectal cancer. RT-PCR was used to amplify the heavy chain Fd and light chain kappa and the amplification products were inserted successively into the vector pComb3 to construct the human libraries of Fab antibodies. They were then panned by phage display technology. By means of Dot immunoblotting and ELISA, the libraries were identified and the Fab phage antibodies binding with antigens of colorectal cancer were selected. RESULTS: The amplified fragments of Fd and kappa gained by RT-PCR were about 650 bp. Fd and kappa PCR products were subsequently inserted into the vector pComb3, resulting in a recombination rate of 40% and the volume of Fab phage display library reached 1.48 x 10(6).The libraries were enriched about 120-fold by 3 cycles of adsorption-elution-multiplication (panning). Dot immunoblotting showed Fab expressions on the phage libraries and ELISA showed 5 clones of Fab phage antibodies which had binding activities with antigens of colorectal cancer. CONCLUSION: The natural immune Fab antibody phage display libraries of colorectal cancer were constructed. They could be used to select the relative antibodies of colorectal cancer.

关键词： genes, immunoglobulin Peptide Library Antibodies Bacteriophages Colorectal Neoplasms Humans immunoglobulin Fab Fragments Research Support, Non-U.S. Gov't

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Cell-specific expression of the diphtheria toxin A-chain coding sequence induces cancer cell suicide

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Chinese Medical Journal 2002年第6期115卷 869-873,152页

作者：芮红兵陈元仲福建省血液病研究所福建医科大学附属协和医院福州350001

OBJECTIVE: To test whether the diphtheria toxin A (DT-A) chain coding sequence linked to murine immunoglobulin Kappa light chain (IgKappa) promoter and enhancer have selective cytocidal effects on IgKappa producing cells. METHODS: The diphtheria toxin A gene or beta galactosidase (beta-gal) gene were linked to a murine IgKappa promoter and enhancer to construct pcDNA3IgKappaDTA or pcDNA3IgKappaLacZ plasmids. These plasmids were transfected into IgKappa producing or non-producing cells by the liposome coated DNA method. Expression of beta-gal activity and effects on cell growth of transfected cells were assessed. RESULTS: The beta-gal gene, under the control of cytomegalovirus (CMV) promoter, can express in all cell lines. Expression of beta-gal under the control of the IgKappa promoter was detected only in the IgKappa producing cell line, CA46. Expression of beta-gal was greatly suppressed when cotransfected with pcDNA3IgKappaDTA in CA46 *** growth of CA46 cells transfected with pcDNA3IgKappaDTA plasmid was significantly inhibited compared with CA46 cells transfected with pcDNA3IgKappaLacZ. CONCLUSION: Selective killing of IgKappa producing cells can be attained by introducing the diphtheria toxin A gene under the control of IgKappa promoter and enhancer.

关键词： genes, immunoglobulin Diphtheria Toxin Enhancer Elements (Genetics) Gene Therapy Humans immunoglobulin kappa-Chains Neoplasms Peptide Fragments Promoter Regions (Genetics) Tumor Cells, Cultured

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