Objective: To construct a fusion gene expression vector of immunoglobulin heavy chain variable region (IgHV) gene fragment and cytokines GM-CSF (granulocyte colony-stimulating factor) or IL-2 (interleukin 2) as a family-specific anti-lymphoma nucleic acid vaccine, and inoculate animals to understand the anti-lymphoma immune function of this vaccine. METHODS: Six IgHV3 fragment gene fragments with widely varying fragment lengths were obtained from a cord blood immunoglobulin gene library, sequenced and then predicted for T-cell epitopes in their heavy chain variable regions using bioinformatic resource analysis, and 16 fragments of IgHV previously constructed in our laboratory were also analyzed. The IgHV1 and IgHV3 genes, which contain the majority of T-cell epitopes, were selected, and the framework region (FR)-dominated IgHV(FR) gene fragment and the fusion gene formed by linking IgHV(FR) to GM-CSF or IL-2 genes were cloned into the eukaryotic expression vector. The expression vectors were transfected with COS cells by liposomes and the expression of GM-CSF or IL-2 was determined by ELISA. Some expression vectors were used as nucleic acid vaccines to immunize mice, and immune responses against lymphoma and normal lymphocytes of the same family were examined by indirect immunofluorescence and cytokine secretion assays. RESULTS: Computer scoring system predictions showed that there were about 30 T-cell epitopes restricted by HLA loci in each IgHV sequence, respectively, and more than 90% of T-cell epitopes were located in the framework region, with the top 10% of T-cell epitopes in the score ranking all located in the framework region. The eukaryotic expression vectors of IgHV1(FR) and IgHV3(FR), which were mainly in the framework region (FR) after removing the CDR3 region, were successfully constructed. The expression of GM-CSF in IgHV(FR)-GM-CSF/pcDNA3.0 was more than 200-fold higher than that of the control; the expression of IL-2 in IgHV(FR)-IL-2/pcDNA3.0 was 60-fold higher than that of the control. Antibodies against the lymphoma Namalwa cell line belonging to the IgHV1 family were detected in the IgHV1(FR)-IL-2 immunized animal group at week 2 after the first immunization, and anti-Namalwa cell antibodies were detected in the IgHV1(FR) group at week 4. The IgHV(FR)-IL-2 group had significantly higher serum IFN-γ levels than the pcDNA3.0 group and the IgHV(FR) group.