The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Dennis D Hickstein - One of the best experts on this subject based on the ideXlab platform.
-
gene therapy for canine leukocyte adhesion deficiency with lentiviral vectors using the murine stem cell virus and human phosphoglycerate kinase promoters
Human Gene Therapy, 2011Co-Authors: Michael J Hunter, Laura M Tuschong, Thomas R Bauer, Tanya H Burkholder, Huifen Zhao, Derek A Persons, Dennis D HicksteinAbstract:Abstract Children with leukocyte adhesion deficiency type 1 (LAD-1) and dogs with canine LAD (CLAD) develop life-threatening bacterial infections due to mutations in the leukocyte integrin CD18. Here, we compared the human phosphoglycerate kinase (hPGK) promoter to the murine stem cell virus (MSCV) promoter/enhancer in a self-inactivating HIV-1–derived lentiviral vector to treat animals with CLAD. Four CLAD dogs were infused with CD34+ cells transduced with the hPGK vector, and two CLAD dogs received MSCV vector–transduced CD34+ cells. Infusions were preceded by a nonmyeloablative dose of 200 cGy total body irradiation. Comparable numbers of transduced cells were infused in each group of animals. Only one of four CLAD animals treated with the hPGK-cCD18 vector had reversal of CLAD, whereas both MSCV-cCD18 vector–treated dogs had reversal of the phenotype. Correction of CLAD depends both upon the percentage of CD18+ myeloid cells and the level of expression of CD18 on individual myeloid cells. In this rega...
-
gene therapy of canine leukocyte adhesion deficiency using lentiviral vectors with human cd11b and CD18 promoters driving canine CD18 expression
Molecular Therapy, 2011Co-Authors: Michael J Hunter, Laura M Tuschong, Cedar J Fowler, Thomas R Bauer, Tanya H Burkholder, Dennis D HicksteinAbstract:To identify cellular promoters in a self-inactivating (SIN) lentiviral vector that might be beneficial in treating children with leukocyte adhesion deficiency type 1 (LAD-1), we tested lentiviral vectors with human CD11 and CD18 leukocyte integrin proximal promoter elements directing expression of canine CD18 in animals with canine LAD (CLAD). Lentiviral vectors with either the human CD11b (637 bp) proximal promoter or the human CD18 (1,060 bp) proximal promoter resulted in the highest percentages of CD18+ CLAD CD34+ cells in vitro. Subsequently, two CLAD dogs were infused with autologous CD34+ cells transduced with the hCD11b (637 bp)-cCD18 vector, and two CLAD dogs were infused with autologous CD34+ cells transduced with the hCD18 (1,060 bp)-cCD18 vector. Each dog received a nonmyeloablative dose of 200 cGy total body irradiation (TBI) before the infusion of transduced cells. The two CLAD dogs treated with the hCD18 (1,060 bp)-cCD18 vector, and one of the two dogs treated with the hCD11b (637 bp)-cCD18 vector, had reversal of the CLAD phenotype. These studies using endogenous leukocyte integrin proximal promoters represent an important step in the development of gene therapy for children with LAD-1.
-
identification of two molecular defects in a child with leukocyte adherence deficiency
Journal of Biological Chemistry, 1992Co-Authors: Anthony L Back, William W Kwok, Dennis D HicksteinAbstract:Children with leukocyte adherence deficiency (LAD), or leukocyte cell adhesion molecule deficiency, experience recurrent, life-threatening bacterial infections related to severe deficiency in surface expression of the leukocyte integrin molecules. The leukocyte integrins consist of a common CD18 (beta) subunit and individual, noncovalently associated alpha subunits designated CD11a, CD11b, and CD11c. Defects in the CD18 subunit prevent surface expression of the CD11/CD18 complexes in children with this disease. We investigated the molecular basis of the disease in a child with the severe deficiency form of LAD and identified two molecular defects in the CD18 subunit. The first defect is a single-base pair C—-T transposition resulting in an amino acid substitution of a leucine for a proline at amino acid 178. This amino acid substitution is located in a region that is highly conserved among the integrin beta subunits and where two previous defects have been located in LAD. The second mutation involves a deletion of 220 base pairs in the cDNA coding for a portion of the extracellular domain and results in a frameshift into a premature stop codon. The deleted region corresponds to a single exon in the CD18 gene. Identification of these two molecular defects in a single child with this disease indicates the compound heterozygous nature of the disorder in this child and identifies regions of the CD18 subunit that may be important for CD11/CD18 heterodimer formation and surface expression.
Michael J Hunter - One of the best experts on this subject based on the ideXlab platform.
-
gene therapy for canine leukocyte adhesion deficiency with lentiviral vectors using the murine stem cell virus and human phosphoglycerate kinase promoters
Human Gene Therapy, 2011Co-Authors: Michael J Hunter, Laura M Tuschong, Thomas R Bauer, Tanya H Burkholder, Huifen Zhao, Derek A Persons, Dennis D HicksteinAbstract:Abstract Children with leukocyte adhesion deficiency type 1 (LAD-1) and dogs with canine LAD (CLAD) develop life-threatening bacterial infections due to mutations in the leukocyte integrin CD18. Here, we compared the human phosphoglycerate kinase (hPGK) promoter to the murine stem cell virus (MSCV) promoter/enhancer in a self-inactivating HIV-1–derived lentiviral vector to treat animals with CLAD. Four CLAD dogs were infused with CD34+ cells transduced with the hPGK vector, and two CLAD dogs received MSCV vector–transduced CD34+ cells. Infusions were preceded by a nonmyeloablative dose of 200 cGy total body irradiation. Comparable numbers of transduced cells were infused in each group of animals. Only one of four CLAD animals treated with the hPGK-cCD18 vector had reversal of CLAD, whereas both MSCV-cCD18 vector–treated dogs had reversal of the phenotype. Correction of CLAD depends both upon the percentage of CD18+ myeloid cells and the level of expression of CD18 on individual myeloid cells. In this rega...
-
gene therapy of canine leukocyte adhesion deficiency using lentiviral vectors with human cd11b and CD18 promoters driving canine CD18 expression
Molecular Therapy, 2011Co-Authors: Michael J Hunter, Laura M Tuschong, Cedar J Fowler, Thomas R Bauer, Tanya H Burkholder, Dennis D HicksteinAbstract:To identify cellular promoters in a self-inactivating (SIN) lentiviral vector that might be beneficial in treating children with leukocyte adhesion deficiency type 1 (LAD-1), we tested lentiviral vectors with human CD11 and CD18 leukocyte integrin proximal promoter elements directing expression of canine CD18 in animals with canine LAD (CLAD). Lentiviral vectors with either the human CD11b (637 bp) proximal promoter or the human CD18 (1,060 bp) proximal promoter resulted in the highest percentages of CD18+ CLAD CD34+ cells in vitro. Subsequently, two CLAD dogs were infused with autologous CD34+ cells transduced with the hCD11b (637 bp)-cCD18 vector, and two CLAD dogs were infused with autologous CD34+ cells transduced with the hCD18 (1,060 bp)-cCD18 vector. Each dog received a nonmyeloablative dose of 200 cGy total body irradiation (TBI) before the infusion of transduced cells. The two CLAD dogs treated with the hCD18 (1,060 bp)-cCD18 vector, and one of the two dogs treated with the hCD11b (637 bp)-cCD18 vector, had reversal of the CLAD phenotype. These studies using endogenous leukocyte integrin proximal promoters represent an important step in the development of gene therapy for children with LAD-1.
Hidetoshi Inoko - One of the best experts on this subject based on the ideXlab platform.
-
modulation of cell surface antigens and regulation of phagocytic activity mediated by cd11b in the monocyte like cell line u937 in response to lipopolysaccharide
Tissue Antigens, 1993Co-Authors: Nabunae Ikewaki, Hidetoshi Inoko, Hidekazu TamauchiAbstract:: Modulation of the cellular antigens and regulation of the phagocytic activity of the monocyte-like cell line U937 after culture with lipopolysaccharide (LPS) were investigated. CD14 expression was induced on the surface of the U937 cells after 48 h of culture with LPS and then they became adhesive with numerous filamentous filopodia extruded on the cell surface, exhibiting the enhanced expression of CD16 and CD23, the activation cell surface markers for differentiation into macrophage. However, no induction or enhancement of the cell surface expression was observed with respect to CD11b, CD18, HLA-A, B, C, HLA-DR, DQ, DP or CD57. These U937 cells also acquired the ability to produce superoxide anions and to phagocytose the Salmonella enteritidis strain, 116–54. This phagocytosis was inhibited by the anti-CD11b monoclonal antibodies, but not by the anti-CD14, anti-CD16, anti-CD18, anti-CD23, anti-HLA-A, B, C or anti-HLA-DR monoclonal antibodies. These findings indicate that the phagocytic activity against Salmonella enteritidis 116–54 induced by LPS is mediated mainly via the CD11b molecule, but is not associated with the increased expression of CD11b. Puromycin and cycloheximide, inhibitors of protein synthesis, or a divalent cation-chelating agent, EDTA completely inhibited this phagocytic activity. Interestingly, EDTA was found to suppress specifically the CD11b expression on the U937 cells cultured with LPS. No phagocytic activity was induced when the U937 cells cultured with LPS were incubated at 4°C, but restored to the control level when shifted up to 37°C. This experimental system provides an excellent model for the investigation of the molecular mechanism underlying modulation of the cell surface antigens and regulation of the phagocytic activity mediated by CD11b in the U937 cells in response to LPS.
Laura M Tuschong - One of the best experts on this subject based on the ideXlab platform.
-
gene therapy for canine leukocyte adhesion deficiency with lentiviral vectors using the murine stem cell virus and human phosphoglycerate kinase promoters
Human Gene Therapy, 2011Co-Authors: Michael J Hunter, Laura M Tuschong, Thomas R Bauer, Tanya H Burkholder, Huifen Zhao, Derek A Persons, Dennis D HicksteinAbstract:Abstract Children with leukocyte adhesion deficiency type 1 (LAD-1) and dogs with canine LAD (CLAD) develop life-threatening bacterial infections due to mutations in the leukocyte integrin CD18. Here, we compared the human phosphoglycerate kinase (hPGK) promoter to the murine stem cell virus (MSCV) promoter/enhancer in a self-inactivating HIV-1–derived lentiviral vector to treat animals with CLAD. Four CLAD dogs were infused with CD34+ cells transduced with the hPGK vector, and two CLAD dogs received MSCV vector–transduced CD34+ cells. Infusions were preceded by a nonmyeloablative dose of 200 cGy total body irradiation. Comparable numbers of transduced cells were infused in each group of animals. Only one of four CLAD animals treated with the hPGK-cCD18 vector had reversal of CLAD, whereas both MSCV-cCD18 vector–treated dogs had reversal of the phenotype. Correction of CLAD depends both upon the percentage of CD18+ myeloid cells and the level of expression of CD18 on individual myeloid cells. In this rega...
-
gene therapy of canine leukocyte adhesion deficiency using lentiviral vectors with human cd11b and CD18 promoters driving canine CD18 expression
Molecular Therapy, 2011Co-Authors: Michael J Hunter, Laura M Tuschong, Cedar J Fowler, Thomas R Bauer, Tanya H Burkholder, Dennis D HicksteinAbstract:To identify cellular promoters in a self-inactivating (SIN) lentiviral vector that might be beneficial in treating children with leukocyte adhesion deficiency type 1 (LAD-1), we tested lentiviral vectors with human CD11 and CD18 leukocyte integrin proximal promoter elements directing expression of canine CD18 in animals with canine LAD (CLAD). Lentiviral vectors with either the human CD11b (637 bp) proximal promoter or the human CD18 (1,060 bp) proximal promoter resulted in the highest percentages of CD18+ CLAD CD34+ cells in vitro. Subsequently, two CLAD dogs were infused with autologous CD34+ cells transduced with the hCD11b (637 bp)-cCD18 vector, and two CLAD dogs were infused with autologous CD34+ cells transduced with the hCD18 (1,060 bp)-cCD18 vector. Each dog received a nonmyeloablative dose of 200 cGy total body irradiation (TBI) before the infusion of transduced cells. The two CLAD dogs treated with the hCD18 (1,060 bp)-cCD18 vector, and one of the two dogs treated with the hCD11b (637 bp)-cCD18 vector, had reversal of the CLAD phenotype. These studies using endogenous leukocyte integrin proximal promoters represent an important step in the development of gene therapy for children with LAD-1.
Tanya H Burkholder - One of the best experts on this subject based on the ideXlab platform.
-
gene therapy for canine leukocyte adhesion deficiency with lentiviral vectors using the murine stem cell virus and human phosphoglycerate kinase promoters
Human Gene Therapy, 2011Co-Authors: Michael J Hunter, Laura M Tuschong, Thomas R Bauer, Tanya H Burkholder, Huifen Zhao, Derek A Persons, Dennis D HicksteinAbstract:Abstract Children with leukocyte adhesion deficiency type 1 (LAD-1) and dogs with canine LAD (CLAD) develop life-threatening bacterial infections due to mutations in the leukocyte integrin CD18. Here, we compared the human phosphoglycerate kinase (hPGK) promoter to the murine stem cell virus (MSCV) promoter/enhancer in a self-inactivating HIV-1–derived lentiviral vector to treat animals with CLAD. Four CLAD dogs were infused with CD34+ cells transduced with the hPGK vector, and two CLAD dogs received MSCV vector–transduced CD34+ cells. Infusions were preceded by a nonmyeloablative dose of 200 cGy total body irradiation. Comparable numbers of transduced cells were infused in each group of animals. Only one of four CLAD animals treated with the hPGK-cCD18 vector had reversal of CLAD, whereas both MSCV-cCD18 vector–treated dogs had reversal of the phenotype. Correction of CLAD depends both upon the percentage of CD18+ myeloid cells and the level of expression of CD18 on individual myeloid cells. In this rega...
-
gene therapy of canine leukocyte adhesion deficiency using lentiviral vectors with human cd11b and CD18 promoters driving canine CD18 expression
Molecular Therapy, 2011Co-Authors: Michael J Hunter, Laura M Tuschong, Cedar J Fowler, Thomas R Bauer, Tanya H Burkholder, Dennis D HicksteinAbstract:To identify cellular promoters in a self-inactivating (SIN) lentiviral vector that might be beneficial in treating children with leukocyte adhesion deficiency type 1 (LAD-1), we tested lentiviral vectors with human CD11 and CD18 leukocyte integrin proximal promoter elements directing expression of canine CD18 in animals with canine LAD (CLAD). Lentiviral vectors with either the human CD11b (637 bp) proximal promoter or the human CD18 (1,060 bp) proximal promoter resulted in the highest percentages of CD18+ CLAD CD34+ cells in vitro. Subsequently, two CLAD dogs were infused with autologous CD34+ cells transduced with the hCD11b (637 bp)-cCD18 vector, and two CLAD dogs were infused with autologous CD34+ cells transduced with the hCD18 (1,060 bp)-cCD18 vector. Each dog received a nonmyeloablative dose of 200 cGy total body irradiation (TBI) before the infusion of transduced cells. The two CLAD dogs treated with the hCD18 (1,060 bp)-cCD18 vector, and one of the two dogs treated with the hCD11b (637 bp)-cCD18 vector, had reversal of the CLAD phenotype. These studies using endogenous leukocyte integrin proximal promoters represent an important step in the development of gene therapy for children with LAD-1.