The bactericidal/permeability increasing protein (BPI) has an important function of nonspecific killing of Gram-negative bacteria. In this study, qPCR was used to detect the expression of the BPI gene in twelve tissue...The bactericidal/permeability increasing protein (BPI) has an important function of nonspecific killing of Gram-negative bacteria. In this study, qPCR was used to detect the expression of the BPI gene in twelve tissues of Meishan piglets from birth to weaning. BPI gene overexpression, bacterial adhesions count and indirect immunofluorescence were applied to analyze the relationship between BPI gene expression and the infectivity of Escherichia coli and Salmonella. The results showed that the BPI gene was expressed highly in duodenum, jejunum and ileum (fold changes of relative expression levels were more than 10 000, 500 and 200, respectively). The expression of the BPI gene at 35 days of age was significantly higher (P<0.01) than that at all other days. Transcription of the BPI gene was up-regulated 2 401-fold in porcine intestinal epithelial (IPEC-J2) cells that were transfected with the BPI gene overexpression lentivirus (IPEC-J2-BPI), and significantly higher (P<0.01) than that in negative control cells (IPEC-J2-NC). Protein expression levels in IPEC-J2-BPI cells were also increased. When IPEC-J2 cells were incubated with E. coli and Salmonella, respectively, for 2, 4, 6, 8, 10 and 12 h, the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.05) than that in IPEC-J2-NC cells. The results of indirect immunofluorescence analysis showed that the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.01) than that in IPEC-J2-NC cells. These results demonstrated that the BPI gene might play an important role in regulating weaning stress especially intestinal-mediated immune response. Overexpression of the BPI gene at the cellular level could significantly enhance the anti-bactericidal ability against Gram-negative bacteria such as E. coli and Salmonella. This has important biological significance in piglet resistance to bacterial diarrhea.展开更多
To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 （AAV2）-mediated BPI-Fcγ1 gene on protecti...To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 （AAV2）-mediated BPI-Fcγ1 gene on protecting mice from challenge of lethal endotoxin. The chimeric BPI-Fcγ1 gene consists of two parts, one encods functional N-terminus （1 to 199 amino acidic residues） of human BPI, which is a bactericidal/permeability-increasing protein, and the other encodes Fc segment of human immunoglobulin G1 （Fcγ1）. Our results indicated that the target protein could be expressed and secreted into the serum of the gene-transferred mice. After lethal endotoxin challenge, the levels of endotoxin and TNF-a in the gene-transferred mice were decreased. The survival rate of the BPI-Fcγ1 gene-transferred mice was markedly increased. Our data suggest that AAV2-mediated chimeric BPI-Fcγ1 gene delivery can potentially be used clinically for the protection and treatment of endotoxemia and endotoxic shock in high-risk individuals. Cellular ＆ Molecular Immunology. 2006;3（3）：221-225.展开更多
In previous research, chimerical BPI23-Fcy1 gene which consisted of human bactericidal/permeability increasing protein （BPI） gene of encoding the functional N terminus （amino acid residues 1 to 199） of human BPI a...In previous research, chimerical BPI23-Fcy1 gene which consisted of human bactericidal/permeability increasing protein （BPI） gene of encoding the functional N terminus （amino acid residues 1 to 199） of human BPI and Fcy1 gene of encoding the Fc segment of human immunoglobulin G1 was successfully reconstructed within a recombinant adeno-associated virus serotype 2 （rAAV2） vector as rAAV2-BPI23-Fcy1. Here, to evaluate the potentiality of applying gene therapy to gram negative bacterial （GNB） infection in high-risk patients, we investigated protection of immuno-compromised mice and immunocompetent mice from challenge with minimal lethal dose （MLD） Klebsiella pneumonia infection after rAAV2-BPI23-Fcy1 gene transferred. The results showed that the survival rate of rAAV2-BPI23-Fcy1 transferred immunocompetent mice as well as immuno-compromised mice （40.0% and 44.4%, respectively） were significant higher than that of corresponding control mice （6.7% and 4.4%, respectively）; the bacteria counting, level of endotoxin and proinflammatory cytokines in the rAAV2-BP123-Fcy1 transferred immuno-compromised mice were markedly lower than that of rAAV2-EGFP and rAAV2-Null transferred immuno- compromised mice. Our data suggest that rAAV2-BPI23-Fcy1 gene transferring offered immuno-compromised mice with resistance against GNB infection, so it is quite potential in preventing GNB infection of clinical high-risk patients. Cellular ＆ Molecular Immunology. 2008;5（6）：439-445.展开更多
基金This study was supported by the National Natural Science Foundation of China(31772560)the Yangzhou University International Academic Exchange Fund(YZUIAEF201901005)+1 种基金the College Students’Innovation and Entrepreneurship Training Program of Jiangsu Province,China(201811117014Z)the Priority Academic Program Development(PAPD)of Jiangsu Higher Education Institutions.
文摘The bactericidal/permeability increasing protein (BPI) has an important function of nonspecific killing of Gram-negative bacteria. In this study, qPCR was used to detect the expression of the BPI gene in twelve tissues of Meishan piglets from birth to weaning. BPI gene overexpression, bacterial adhesions count and indirect immunofluorescence were applied to analyze the relationship between BPI gene expression and the infectivity of Escherichia coli and Salmonella. The results showed that the BPI gene was expressed highly in duodenum, jejunum and ileum (fold changes of relative expression levels were more than 10 000, 500 and 200, respectively). The expression of the BPI gene at 35 days of age was significantly higher (P<0.01) than that at all other days. Transcription of the BPI gene was up-regulated 2 401-fold in porcine intestinal epithelial (IPEC-J2) cells that were transfected with the BPI gene overexpression lentivirus (IPEC-J2-BPI), and significantly higher (P<0.01) than that in negative control cells (IPEC-J2-NC). Protein expression levels in IPEC-J2-BPI cells were also increased. When IPEC-J2 cells were incubated with E. coli and Salmonella, respectively, for 2, 4, 6, 8, 10 and 12 h, the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.05) than that in IPEC-J2-NC cells. The results of indirect immunofluorescence analysis showed that the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.01) than that in IPEC-J2-NC cells. These results demonstrated that the BPI gene might play an important role in regulating weaning stress especially intestinal-mediated immune response. Overexpression of the BPI gene at the cellular level could significantly enhance the anti-bactericidal ability against Gram-negative bacteria such as E. coli and Salmonella. This has important biological significance in piglet resistance to bacterial diarrhea.
文摘To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 （AAV2）-mediated BPI-Fcγ1 gene on protecting mice from challenge of lethal endotoxin. The chimeric BPI-Fcγ1 gene consists of two parts, one encods functional N-terminus （1 to 199 amino acidic residues） of human BPI, which is a bactericidal/permeability-increasing protein, and the other encodes Fc segment of human immunoglobulin G1 （Fcγ1）. Our results indicated that the target protein could be expressed and secreted into the serum of the gene-transferred mice. After lethal endotoxin challenge, the levels of endotoxin and TNF-a in the gene-transferred mice were decreased. The survival rate of the BPI-Fcγ1 gene-transferred mice was markedly increased. Our data suggest that AAV2-mediated chimeric BPI-Fcγ1 gene delivery can potentially be used clinically for the protection and treatment of endotoxemia and endotoxic shock in high-risk individuals. Cellular ＆ Molecular Immunology. 2006;3（3）：221-225.
基金funded by the Beijing Municipal Natural Science Foundation (7082016)
文摘In previous research, chimerical BPI23-Fcy1 gene which consisted of human bactericidal/permeability increasing protein （BPI） gene of encoding the functional N terminus （amino acid residues 1 to 199） of human BPI and Fcy1 gene of encoding the Fc segment of human immunoglobulin G1 was successfully reconstructed within a recombinant adeno-associated virus serotype 2 （rAAV2） vector as rAAV2-BPI23-Fcy1. Here, to evaluate the potentiality of applying gene therapy to gram negative bacterial （GNB） infection in high-risk patients, we investigated protection of immuno-compromised mice and immunocompetent mice from challenge with minimal lethal dose （MLD） Klebsiella pneumonia infection after rAAV2-BPI23-Fcy1 gene transferred. The results showed that the survival rate of rAAV2-BPI23-Fcy1 transferred immunocompetent mice as well as immuno-compromised mice （40.0% and 44.4%, respectively） were significant higher than that of corresponding control mice （6.7% and 4.4%, respectively）; the bacteria counting, level of endotoxin and proinflammatory cytokines in the rAAV2-BP123-Fcy1 transferred immuno-compromised mice were markedly lower than that of rAAV2-EGFP and rAAV2-Null transferred immuno- compromised mice. Our data suggest that rAAV2-BPI23-Fcy1 gene transferring offered immuno-compromised mice with resistance against GNB infection, so it is quite potential in preventing GNB infection of clinical high-risk patients. Cellular ＆ Molecular Immunology. 2008;5（6）：439-445.
