Bovine Herpesvirus-1 （BoHV-1） is a DNA virus belonging to the family Herpesviridae, subfamily Alfaherpesvirinae; it is a worldwide pathogen, causing serious economic losses in livestock. In Colombia there have been ...Bovine Herpesvirus-1 （BoHV-1） is a DNA virus belonging to the family Herpesviridae, subfamily Alfaherpesvirinae; it is a worldwide pathogen, causing serious economic losses in livestock. In Colombia there have been multiple isolates of BoHV-1 that have been subjected to molecular characterization, classifying most of the country isolates as BoHV-I.1. In the present study we developed and evaluated an ethyleneimine binary inactivated isolate from the native BoHV-1 strain （C6rdoba-2） in a rabbit model of vaccination and infection. The vaccine was evaluated in two phases, one of immunogenicity with vaccination and a booster after 21 days, and an evaluation phase of protection against challenge with a highly virulent reference strain. The results demonstrate optimum serum-conversion, with protective neutralizing antibody titers 28 days post vaccination and optimal protection against challenge with the reference strain with decreased clinical signs of infection, protection against the onset of fever and decrease of virus excretion post challenge. In conclusion, our results show the enormous potential that an immunogenic inactivated vaccine has produced from the native BoHV-I.1 strain, which produces a high antigen mass to the vaccine to induce optimal immunity and protection, and it is a strong candidate for evaluation and possible future use in different cattle populations.展开更多
Tumor cell-derived exosomes have been proposed as non-cellular nanomeric vaccine which could induce potent anti- tumor immune response in mice. In order to develop the protocols to prepare tumor cell-derived exosomes ...Tumor cell-derived exosomes have been proposed as non-cellular nanomeric vaccine which could induce potent anti- tumor immune response in mice. In order to develop the protocols to prepare tumor cell-derived exosomes for basic research and clinical trail, we isolated exosomes from ovalbumin (OVA)-expressing thymoma cells EG.7-OVA by various preparation methods. We demonstrate the non-sedimentation method is simple, rapid, efficient with higher yield and purity of exosomes. EG.7-OVA-derived exosomes are 40-100 nm in diameter sequestered by lipid bi-layer, and contain rich heat shock protein (HSP) and OVA. The result of the size distribution determination is consistent with the calculation by the visual microscopic inspection, with 90.4% particles at the range of 50-90 nm. Moreover, as a model antigen of the EG.7 cells, OVA concentra- tion in EG.7-derived exosomes can be regarded as a good quality control parameter. Therefore, we have established a platform to efficiently prepare exosomes for tumor immunotherapy.展开更多
The antigenic determinants of trichosanthin were predicted by molecular modeling. First, the threedimensional structure model of the antigen-binding fragment of anti-trichosanthin immunoglobulin E was built on the bas...The antigenic determinants of trichosanthin were predicted by molecular modeling. First, the threedimensional structure model of the antigen-binding fragment of anti-trichosanthin immunoglobulin E was built on the basis of its amino-acid sequence and the known three-dimensional structure of an antibody with similar sequence. Secondly, the preferable antigen-antibody interactions were obtained based on the known three-dimensional structure of trichosanthin and of the hypervariable regions of anti-trichosanthin immunoglobulin E. Two regions in the molecular surface of trichosanthin were found to form extensive interactions with the hypervariable regions of the antibody and have been predicted to be the possible antigenic determinants: one is composed of two polypeptide segments, Ile201-Glu210 and Ile225-Asp229, which are close to each other in the three-dimensional structure; and the other is the segment Lys173-Thr178. The former region seems to be the more reasonable antigenic determinant than the latter one.展开更多
To find out the protective polypeptide epitopes of HCV HVR1, the antigenicity of the synthetic peptide was predicted by computer modeling and verified by ELISA and lymphocyte transformation test. It was found that the...To find out the protective polypeptide epitopes of HCV HVR1, the antigenicity of the synthetic peptide was predicted by computer modeling and verified by ELISA and lymphocyte transformation test. It was found that the outcome of the computer modeling was in accord with the experimental results. The method by using computer modeling would be a economic approach by which the protective peptides could be identified quickly.展开更多
OBJECTIVE:To study the effect of Lichong Decoction(Lichong Decoction for strengthening anti-pathogenic Qi and eliminating blood stasis) on the expression of insulin-like growth factor-I(IGF-I) and proliferating cell n...OBJECTIVE:To study the effect of Lichong Decoction(Lichong Decoction for strengthening anti-pathogenic Qi and eliminating blood stasis) on the expression of insulin-like growth factor-I(IGF-I) and proliferating cell nuclear antigen(PCNA) mRNA in a rat model of uterine leiomyoma.METHODS:Fifty female Wistar rats were randomized into a normal control group,model group,Lichong Decoction group,Guizhifuling Capsule(Capsule containing Cassia Twig and Poria) group,and Mifepristone group.The uterine leiomyoma model was established by peritoneal injections of exogenous estrogen and progesterone hormone.The ultrastructural changes in cells of rat uterine tissues were observed with transmission electron microscopy,and the expression of IGF-I and PCNA mRNA was detected by real-time fluorescent quantitative PCR.RESULTS:Following treatment,cells in the Lichong Decoction group appeared to be arranged normally,the cellular morphology were almost in a normal state,hyperplasia and hypertrophy of the chondriosome was reduced,collagen fibers were arranged in a regular manner,without obvious hyperplasia,and the expression of IGF-I and PCNA mRNA was significantly decreased compared with the model group(P<0.01).CONCLUSIONS:The effect of Lichong Decoction on uterine leiomyoma is related to its function in reducing the expression of IGF-I and PCNA mRNA.展开更多
SAMHD1(Sterile Alpha Motif and Histidine-aspartate Domain containing protein 1) has been documented as a host factor that restricts HIV-1 and some DNA viruses. In this work, we attempted to explore possible effects of...SAMHD1(Sterile Alpha Motif and Histidine-aspartate Domain containing protein 1) has been documented as a host factor that restricts HIV-1 and some DNA viruses. In this work, we attempted to explore possible effects of SAMHD1 on exogenous DNA and show that SAMHD1 exerts a general inhibition on the expression of exogenous DNA in vitro and in mice. This inhibition is achieved through repressing transcription of exogenous DNA. Intriguingly, unlike SAMHD1’s restriction of HIV-1, such restriction does not require the dNTPase or RNase activities, or T592 phosphorylation of SAMHD1. Mechanistically,SAMHD1 enhances the expression of interferon regulatory factor-1(IRF1), while IRF1 upregulation was demonstrated to inhibit exogenous DNA expression in a similar fashion as SAMHD1. IFNk1, whose induction has been associated with IRF1 activation, is dispensable for SAMHD1/IRF1-mediated restriction of exogenous DNA, and neither type Ⅰ nor Ⅱ interferons appear to be involved. We also demonstrate that SAMHD1/IRF1-mediated restriction can effectively inhibit hepatitis B virus(HBV) antigen expression and progeny virus production in mouse models. In conclusion, these data support restriction of exogenous DNA as a novel function of SAMHD1.展开更多
Recently, genome wide association studies showed that there is a strong association between abacavir-induced serious, idio- syncratic, adverse drag reactions （ADRs） and human leukocyte antigen-B＊5701 （HLA-B＊5701...Recently, genome wide association studies showed that there is a strong association between abacavir-induced serious, idio- syncratic, adverse drag reactions （ADRs） and human leukocyte antigen-B＊5701 （HLA-B＊5701）. Studies also found that ab- acavir-induced ADRs were seldom observed in patients carrying the HLA-B＊5801 subtype. HLA-B＊5801 of the same sero- type （B17） as B＊5701 differs by only 4 amino acids from B＇5701. It is believed that because of these sequence differences, HLA-B＊5801 cannot bind the specific peptides which are required for HLA-B＊5701 to stimulate the T cell immune response. Thus, the difference in peptide binding profiles between HLA-B＊5701 and B＊5801 is an important clue for exploring the mechanisms of abacavir-induced ADRs. VHSE （principal component score vector of hydrophobic, steric, and electronic prop- erties）, a set of amino acid structural descriptors, was employed to establish QSAR models of peptide-binding affinities of HLA-B＊5701 and B＊5801. Optimal linear SVM （support vector machine） models with high predictive capabilities were ob- tained for both B＊5701 and B＇5801. The R2 （coefficient of determination）, Q2 （cross-validated R：）, and RpRE2 （R2 of test set） of two optimal models were 0,7530, 0.7037, 0.6153 （B＇5701） and 0.6074, 0.5966, 0.5762 （B＊5801）, respectively. For B＇5701 and B＇5801, the mutations in positions 45 （MET-THR） and 46 （ALA-GLU） have little influence on the selection specificity of the P2 position of the bound peptide. However, the mutation in position 97 （VAL-ARG） greatly influences the selection speci- ficity of the P7 position. HLA-B＊5701 prefers the bulky and positively charged amino acids at the P7 position. In contrast, HLA-B＊5801 prefers the non-polar hydrophobic amino acids at the P7 position while positively charged amino acids are un- favored.展开更多
基金supported by the División de Investigación Universidad Nacional de Colombia, grants No.20201007738 and 202010013254
文摘Bovine Herpesvirus-1 （BoHV-1） is a DNA virus belonging to the family Herpesviridae, subfamily Alfaherpesvirinae; it is a worldwide pathogen, causing serious economic losses in livestock. In Colombia there have been multiple isolates of BoHV-1 that have been subjected to molecular characterization, classifying most of the country isolates as BoHV-I.1. In the present study we developed and evaluated an ethyleneimine binary inactivated isolate from the native BoHV-1 strain （C6rdoba-2） in a rabbit model of vaccination and infection. The vaccine was evaluated in two phases, one of immunogenicity with vaccination and a booster after 21 days, and an evaluation phase of protection against challenge with a highly virulent reference strain. The results demonstrate optimum serum-conversion, with protective neutralizing antibody titers 28 days post vaccination and optimal protection against challenge with the reference strain with decreased clinical signs of infection, protection against the onset of fever and decrease of virus excretion post challenge. In conclusion, our results show the enormous potential that an immunogenic inactivated vaccine has produced from the native BoHV-I.1 strain, which produces a high antigen mass to the vaccine to induce optimal immunity and protection, and it is a strong candidate for evaluation and possible future use in different cattle populations.
文摘Tumor cell-derived exosomes have been proposed as non-cellular nanomeric vaccine which could induce potent anti- tumor immune response in mice. In order to develop the protocols to prepare tumor cell-derived exosomes for basic research and clinical trail, we isolated exosomes from ovalbumin (OVA)-expressing thymoma cells EG.7-OVA by various preparation methods. We demonstrate the non-sedimentation method is simple, rapid, efficient with higher yield and purity of exosomes. EG.7-OVA-derived exosomes are 40-100 nm in diameter sequestered by lipid bi-layer, and contain rich heat shock protein (HSP) and OVA. The result of the size distribution determination is consistent with the calculation by the visual microscopic inspection, with 90.4% particles at the range of 50-90 nm. Moreover, as a model antigen of the EG.7 cells, OVA concentra- tion in EG.7-derived exosomes can be regarded as a good quality control parameter. Therefore, we have established a platform to efficiently prepare exosomes for tumor immunotherapy.
文摘The antigenic determinants of trichosanthin were predicted by molecular modeling. First, the threedimensional structure model of the antigen-binding fragment of anti-trichosanthin immunoglobulin E was built on the basis of its amino-acid sequence and the known three-dimensional structure of an antibody with similar sequence. Secondly, the preferable antigen-antibody interactions were obtained based on the known three-dimensional structure of trichosanthin and of the hypervariable regions of anti-trichosanthin immunoglobulin E. Two regions in the molecular surface of trichosanthin were found to form extensive interactions with the hypervariable regions of the antibody and have been predicted to be the possible antigenic determinants: one is composed of two polypeptide segments, Ile201-Glu210 and Ile225-Asp229, which are close to each other in the three-dimensional structure; and the other is the segment Lys173-Thr178. The former region seems to be the more reasonable antigenic determinant than the latter one.
基金This study was supported by 95 military program ( No. 96M17498Z089)
文摘To find out the protective polypeptide epitopes of HCV HVR1, the antigenicity of the synthetic peptide was predicted by computer modeling and verified by ELISA and lymphocyte transformation test. It was found that the outcome of the computer modeling was in accord with the experimental results. The method by using computer modeling would be a economic approach by which the protective peptides could be identified quickly.
基金Supported by the National Natural Science Foundation of China(No.81073096)the Natural Science Foundation of Beijing City(No.7082015)the "Cultivation Plan for Youth Backbone Talents" Project(PHR 201008403) of Higher SchoolTalent Strong Education Plan
文摘OBJECTIVE:To study the effect of Lichong Decoction(Lichong Decoction for strengthening anti-pathogenic Qi and eliminating blood stasis) on the expression of insulin-like growth factor-I(IGF-I) and proliferating cell nuclear antigen(PCNA) mRNA in a rat model of uterine leiomyoma.METHODS:Fifty female Wistar rats were randomized into a normal control group,model group,Lichong Decoction group,Guizhifuling Capsule(Capsule containing Cassia Twig and Poria) group,and Mifepristone group.The uterine leiomyoma model was established by peritoneal injections of exogenous estrogen and progesterone hormone.The ultrastructural changes in cells of rat uterine tissues were observed with transmission electron microscopy,and the expression of IGF-I and PCNA mRNA was detected by real-time fluorescent quantitative PCR.RESULTS:Following treatment,cells in the Lichong Decoction group appeared to be arranged normally,the cellular morphology were almost in a normal state,hyperplasia and hypertrophy of the chondriosome was reduced,collagen fibers were arranged in a regular manner,without obvious hyperplasia,and the expression of IGF-I and PCNA mRNA was significantly decreased compared with the model group(P<0.01).CONCLUSIONS:The effect of Lichong Decoction on uterine leiomyoma is related to its function in reducing the expression of IGF-I and PCNA mRNA.
基金This work was supported by National Natural Science Foundation of China(81472226,81971921 and 31670166)National Key Project for Infectious Diseases of China(2017ZX10202202 and 2018ZX10301208)+1 种基金Chinese Academy of Medical Sciences(2018PT31044)Shanghai Municipal Education Commission(2017-01-07-00-07-E00057).
文摘SAMHD1(Sterile Alpha Motif and Histidine-aspartate Domain containing protein 1) has been documented as a host factor that restricts HIV-1 and some DNA viruses. In this work, we attempted to explore possible effects of SAMHD1 on exogenous DNA and show that SAMHD1 exerts a general inhibition on the expression of exogenous DNA in vitro and in mice. This inhibition is achieved through repressing transcription of exogenous DNA. Intriguingly, unlike SAMHD1’s restriction of HIV-1, such restriction does not require the dNTPase or RNase activities, or T592 phosphorylation of SAMHD1. Mechanistically,SAMHD1 enhances the expression of interferon regulatory factor-1(IRF1), while IRF1 upregulation was demonstrated to inhibit exogenous DNA expression in a similar fashion as SAMHD1. IFNk1, whose induction has been associated with IRF1 activation, is dispensable for SAMHD1/IRF1-mediated restriction of exogenous DNA, and neither type Ⅰ nor Ⅱ interferons appear to be involved. We also demonstrate that SAMHD1/IRF1-mediated restriction can effectively inhibit hepatitis B virus(HBV) antigen expression and progeny virus production in mouse models. In conclusion, these data support restriction of exogenous DNA as a novel function of SAMHD1.
基金supported by the National Natural Science Foundation of China (Grant No. 61073135)Chongqing Natural Science Foundation(Grant No. CSTC, 2009BA5068)
文摘Recently, genome wide association studies showed that there is a strong association between abacavir-induced serious, idio- syncratic, adverse drag reactions （ADRs） and human leukocyte antigen-B＊5701 （HLA-B＊5701）. Studies also found that ab- acavir-induced ADRs were seldom observed in patients carrying the HLA-B＊5801 subtype. HLA-B＊5801 of the same sero- type （B17） as B＊5701 differs by only 4 amino acids from B＇5701. It is believed that because of these sequence differences, HLA-B＊5801 cannot bind the specific peptides which are required for HLA-B＊5701 to stimulate the T cell immune response. Thus, the difference in peptide binding profiles between HLA-B＊5701 and B＊5801 is an important clue for exploring the mechanisms of abacavir-induced ADRs. VHSE （principal component score vector of hydrophobic, steric, and electronic prop- erties）, a set of amino acid structural descriptors, was employed to establish QSAR models of peptide-binding affinities of HLA-B＊5701 and B＊5801. Optimal linear SVM （support vector machine） models with high predictive capabilities were ob- tained for both B＊5701 and B＇5801. The R2 （coefficient of determination）, Q2 （cross-validated R：）, and RpRE2 （R2 of test set） of two optimal models were 0,7530, 0.7037, 0.6153 （B＇5701） and 0.6074, 0.5966, 0.5762 （B＊5801）, respectively. For B＇5701 and B＇5801, the mutations in positions 45 （MET-THR） and 46 （ALA-GLU） have little influence on the selection specificity of the P2 position of the bound peptide. However, the mutation in position 97 （VAL-ARG） greatly influences the selection speci- ficity of the P7 position. HLA-B＊5701 prefers the bulky and positively charged amino acids at the P7 position. In contrast, HLA-B＊5801 prefers the non-polar hydrophobic amino acids at the P7 position while positively charged amino acids are un- favored.
