AIM: To establish a method for induced and amplified porcine monocyte-derived immature dendritic cells(imDCs).The aim of this study was to assess the pCTLA4-Ig gene-modified porcine imDCs could benefit for prevent the proliferation of xenogeneic spleen cells. METHODS: Poricne peripheral blood mononuclear cells(PBMC) were cultured with rpGM- CSF(30 microg/L) and rpIL-4(20 microg/L). To evaluate the suspending cells by transmission electronic microscope and flow cytometer. Porcine imDCs were transduced with adenovirus mediated pCTLA4-Ig gene. The mixed lymphocyte reaction(MLR) was used to examine the proliferation of spleen cells of SD rat to gene-modified imDCs. RESULTS: With rpGM-CSF and rpIL-4 induced porcine PBMC for 4-5 days, it was exhibited typical morphological characteristics and immunological phenotype of imDCs with high expression of SLA-DR, CD172a and low expression of CD80/CD86 on the cellular surface. The gene-modified imDCs were able to amplified the pCTLA4-Ig and IDO gene by RT-PCR. The stimulate index of gene-modified imDCs group were markedly inferior to that in unmodified imDCs group(P<0.05). CONCLUSION: The pCTLA4-Ig gene-modified imDCs can significantly inhibit the proliferation of xenogeneic spleen cells, and there might was correlated with the expression of IDO reduce the immune response of xenogeneic antigen.
AIM: To establish a method for induced and amplified porcine monocyte-derived immature dendritic cells(imDCs).The aim of this study was to assess the pCTLA4-Ig gene-modified porcine imDCs could benefit for prevent the proliferation of xenogeneic spleen cells. METHODS: Poricne peripheral blood mononuclear cells(PBMC) were cultured with rpGM- CSF(30 microg/L) and rpIL-4(20 microg/L). To evaluate the suspending cells by transmission electronic microscope and flow cytometer. Porcine imDCs were transduced with adenovirus mediated pCTLA4-Ig gene. The mixed lymphocyte reaction(MLR) was used to examine the proliferation of spleen cells of SD rat to gene-modified imDCs. RESULTS: With rpGM-CSF and rpIL-4 induced porcine PBMC for 4-5 days, it was exhibited typical morphological characteristics and immunological phenotype of imDCs with high expression of SLA-DR, CD172a and low expression of CD80/CD86 on the cellular surface. The gene-modified imDCs were able to amplified the pCTLA4-Ig and IDO gene by RT-PCR. The stimulate index of gene-modified imDCs group were markedly inferior to that in unmodified imDCs group(P<0.05). CONCLUSION: The pCTLA4-Ig gene-modified imDCs can significantly inhibit the proliferation of xenogeneic spleen cells, and there might was correlated with the expression of IDO reduce the immune response of xenogeneic antigen.