Whether the immunostimulatory effects of CpG-oligonucleotides （CpG ODNs） could be enhanced by the use of gold nanoparticles （Au-NP） was investigated.The CpG ODNs were modified by the Au-NP （CpG /Au-NP） and their...Whether the immunostimulatory effects of CpG-oligonucleotides （CpG ODNs） could be enhanced by the use of gold nanoparticles （Au-NP） was investigated.The CpG ODNs were modified by the Au-NP （CpG /Au-NP） and their uptake and distribution in murine N9 microglial cells were studied.The immunostimulatory effects of CpG /Au-NP on N9 cells,human B cells and plamacytoid dendritic cells （pDCs） were examined.Results showed that the uptake of CpG /Au-NP in N9 cells was much higher than that of CpG ODNs and CpG /Au-NP localized in the cytoplasm of N9 cells.The amount of TNF-α and IL12p40 in N9 cells was increased greatly by the use of Au-NP.And the amount of IL-6 in B cells and IFN-α in pDCs was also significantly increased,while the activation of B cells was not changed.These results reveal that the Au-NP can be used as a delivery media for the oligonucleotides-based therapeutics.展开更多
Objective To study the regulating effects of a novel CpG oligodeoxynuleotide and the synergistic effect of chitosan-nanoparticles （CNP） with CpG on immune responses of mice, which were used to develop a novel immuno...Objective To study the regulating effects of a novel CpG oligodeoxynuleotide and the synergistic effect of chitosan-nanoparticles （CNP） with CpG on immune responses of mice, which were used to develop a novel immunoadjuvant to boost immune response to conventional vaccines. Methods A novel CpG ODN containing 11 CpG motifs was synthesized and its bioactivities to stimulate the proliferation of lymphocytes of pig in vitro were detected. Then it was entrapped with CNP prepared in our laboratory by the method of ionic cross linkage, and immunized Kunming mice were co-inoculated with paratyphoid vaccine. The peripheral blood was collected weekly from the tail vein of inoculated mice to detect the contents of IgG, IgA, IgM, and specific antibody against salmonella as well as the levels of interleukin-2 （IL2）, IL-4, and IL-6 by SABC-ELISA assay. The numbers of leucocytes, monocytes, granuloytes, and lymphocytes were calculated separately using the routine method. The experimental mice were orally challenged with virulent salmonella 35 days after inoculation. Results This CpG ODN could remarkably provoke the proliferation of lymphocytes of pig in vitro in contrast with the control （P〈0.05）. Compared with those of the control, immunoglobulins, including IgG, IgA, IgM, and specific antibodies to paratyphoid vaccine, increased significantly in sera from the CpG or CpG-CNP-vaccinated mice （P〈0.05）. IL-2, IL-4, and IL-6 increased remarkably in sera from immunized mice （P〈0.05）. The leucocytes, monocytes, granuloytes, and lymphocytes of the mice immunized with CpG or CpG-CNP were also increased in number （P〈0.05）. After the challenge, these immunity values were elevated in the mice vaccinated with CpG or CpG-CNP. The immunized mice all survived, while the control mice fell ill with evident lesions with diffuse hemorrhage in stomach, small intestine, and peritoneum. Conclusions CpG ODN entrapped with CNP is a promising effective immunoadjuvant for vaccination, which promotes humoral and cellular 展开更多
基金Supported by the National Natural Science Foundation of China(No.20807017)Doctoral Fund of Ministry of Education of China (No.20080487087)the Fundamental Research Funds for the Central Universities (No.2010MS091)
文摘Whether the immunostimulatory effects of CpG-oligonucleotides （CpG ODNs） could be enhanced by the use of gold nanoparticles （Au-NP） was investigated.The CpG ODNs were modified by the Au-NP （CpG /Au-NP） and their uptake and distribution in murine N9 microglial cells were studied.The immunostimulatory effects of CpG /Au-NP on N9 cells,human B cells and plamacytoid dendritic cells （pDCs） were examined.Results showed that the uptake of CpG /Au-NP in N9 cells was much higher than that of CpG ODNs and CpG /Au-NP localized in the cytoplasm of N9 cells.The amount of TNF-α and IL12p40 in N9 cells was increased greatly by the use of Au-NP.And the amount of IL-6 in B cells and IFN-α in pDCs was also significantly increased,while the activation of B cells was not changed.These results reveal that the Au-NP can be used as a delivery media for the oligonucleotides-based therapeutics.
文摘Objective To study the regulating effects of a novel CpG oligodeoxynuleotide and the synergistic effect of chitosan-nanoparticles （CNP） with CpG on immune responses of mice, which were used to develop a novel immunoadjuvant to boost immune response to conventional vaccines. Methods A novel CpG ODN containing 11 CpG motifs was synthesized and its bioactivities to stimulate the proliferation of lymphocytes of pig in vitro were detected. Then it was entrapped with CNP prepared in our laboratory by the method of ionic cross linkage, and immunized Kunming mice were co-inoculated with paratyphoid vaccine. The peripheral blood was collected weekly from the tail vein of inoculated mice to detect the contents of IgG, IgA, IgM, and specific antibody against salmonella as well as the levels of interleukin-2 （IL2）, IL-4, and IL-6 by SABC-ELISA assay. The numbers of leucocytes, monocytes, granuloytes, and lymphocytes were calculated separately using the routine method. The experimental mice were orally challenged with virulent salmonella 35 days after inoculation. Results This CpG ODN could remarkably provoke the proliferation of lymphocytes of pig in vitro in contrast with the control （P〈0.05）. Compared with those of the control, immunoglobulins, including IgG, IgA, IgM, and specific antibodies to paratyphoid vaccine, increased significantly in sera from the CpG or CpG-CNP-vaccinated mice （P〈0.05）. IL-2, IL-4, and IL-6 increased remarkably in sera from immunized mice （P〈0.05）. The leucocytes, monocytes, granuloytes, and lymphocytes of the mice immunized with CpG or CpG-CNP were also increased in number （P〈0.05）. After the challenge, these immunity values were elevated in the mice vaccinated with CpG or CpG-CNP. The immunized mice all survived, while the control mice fell ill with evident lesions with diffuse hemorrhage in stomach, small intestine, and peritoneum. Conclusions CpG ODN entrapped with CNP is a promising effective immunoadjuvant for vaccination, which promotes humoral and cellular
