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Intralymphatic Immunotherapy and Vaccination in Mice
Published on: February 2, 2014
[The experimental study on the idiotypic nucleic Acid vaccine constructed from the genomic DNA to lymphoma]
Ning-Jing Lin1, Ping Zhu, Ye-Ping Zhang
1Department of Hematology, The First Hospital of Peking University, Beijing 100034, China.
This study was to investigate the anti-lymphocytic malignancy immunologic effects induced by two types of the idiotypic nucleic acid vaccines which were constructed from the genomic DNA and RNA of the human B lymphoma cell line respectively. Namalwa cell line and BALB/c mice were used as the models. The gene fragments of the IgH variable region (IgHV), which were obtained from the genomic DNA and RNA of Namalwa cell respectively, were cloned into the eukaryocytic expression vector pcDNA 3.0 to be used as the idiotypic nucleic acid vaccines. After transfecting COS cells with one of vaccines constructed from the genomic DNA by using LipofectAMINE, the result of transcription was identified by using RT-PCR. The experimental mice were immunized by intramuscular injection with two types of vaccines. The specific anti-idiotypic antibody was detected by indirect immunofluorescence assay. The results showed that the nucleic acid vaccine constructed from the genomic DNA can be transcribed in COS cells, the transcription product turned shorter, and the intron region of 86 bp was spliced accurately. When immunizing the mice, two vaccines both induced the anti-idiotypic antibody against Namalwa cell, the anti-idiotypic antibody could be detected since detected since after immunization, and got to the peak of titer on the sixth week. It was concluded that the nucleic acid vaccines against lymphoma can be constructed from both the genomic DNA and RNA.
This study was to investigate the anti-lymphocytic malignancy immunologic effects induced by two types of the idiotypic nucleic acid vaccines which were constructed from the genomic DNA and RNA of the human B lymphoma cell line respectively. Namalwa cell line and BALB/c mice were used as the models. The gene fragments of the IgH variable region (IgHV), which were obtained from the genomic DNA and RNA of Namalwa cell respectively, were cloned into the eukaryocytic expression vector pcDNA 3.0 to be used as the idiotypic nucleic acid vaccines. After transfecting COS cells with one of vaccines constructed from the genomic DNA by using LipofectAMINE, the result of transcription was identified by using RT-PCR. The experimental mice were immunized by intramuscular injection with two types of vaccines. The specific anti-idiotypic antibody was detected by indirect immunofluorescence assay. The results showed that the nucleic acid vaccine constructed from the genomic DNA can be transcribed in COS cells, the transcription product turned shorter, and the intron region of 86 bp was spliced accurately. When immunizing the mice, two vaccines both induced the anti-idiotypic antibody against Namalwa cell, the anti-idiotypic antibody could be detected since detected since after immunization, and got to the peak of titer on the sixth week. It was concluded that the nucleic acid vaccines against lymphoma can be constructed from both the genomic DNA and RNA.
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