Drug-selected complete restoration of superoxide generation in Epstein-Barr virus-transformed B cells from p47 phox -deficient chronic granulomatous disease patients by using a bicistronic retrovirus vector encoding a human multi-drug resistance gene ( MDR 1) and the p47 phox gene

Abstract Patients with chronic granulomatous disease (CGD) have recurrent infections resulting from a failure of phagocytic cells to produce superoxide. One third of CGD patients have an autosomal gene defect resulting in absence of p47phox protein, a cytoplasmic component critical to superoxide production by phagocytic cells. cDNA encoding p47phox has been cloned and recombinant p47phox (rp47phox) restores superoxide-generating activity to a cell-free assay containing cell membranes and cytosol from p47phox-deficient CGD neutrophils. The goal of the present study was to determine the feasibility of retrovirus mediated expression of rp47phox in the HL60 and U937 human hematopoietic cell lines, and in an Epstein-Barr virus transformed B- lymphocyte cell line (EBV-BCL) derived from a p47phox-deficient CGD patient. Normal EBV-BCL contain p47phox and generate small amounts of superoxide, while this CGD EBV-BCL lacks any detectable p47phox protein. Defective amphotropic retrovirus containing p47phox sequence inserted in the LXSN vector in sense and antisense orientations were used to transduce HL60, U937, and CGD EBV-BCL. p47phox mRNA sequence was detected in cells transduced with either sense or antisense retroviral constructs while rp47phox protein was detected only with the sense construct. The amount of rp47phox protein produced within these cells was greater than the native p47phox present in uninduced HL60 or U937 cells, but substantially less than that present in normal neutrophils, induced HL60 cells, or even normal EBV-BCL. Differentiation of transduced HL60 cells and the associated production of native p47phox in response to dimethyl sulfoxide was not affected. These studies demonstrate that retrovirus constructs can be used to mediate stable expression of rp47phox protein in human hematopoietic cell lines and can restore rp47phox protein within the cytosol of p47phox-deficient EBV-BCL from patients with CGD.

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Epstein-Barr Virus–Negative Granulomatous Disease Due to SAP Deficiency

Journal of Clinical Immunology ◽

10.1007/s10875-021-01032-4 ◽

2021 ◽

Author(s):

Takao Karasawa ◽

Ko Kudo ◽

Kay Tanita ◽

Yoshihiro Takahashi ◽

Hirokazu Kanegane ◽

...

Keyword(s):

Epstein Barr Virus ◽

Granulomatous Disease ◽

Barr Virus ◽

Epstein Barr

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Single-Step Conversion of Cells to Retrovirus Vector Producers with Herpes Simplex Virus–Epstein-Barr Virus Hybrid Amplicons

Journal of Virology ◽

10.1128/jvi.73.12.10426-10439.1999 ◽

1999 ◽

Vol 73 (12) ◽

pp. 10426-10439 ◽

Cited By ~ 56

Author(s):

Miguel Sena-Esteves ◽

Yoshinaga Saeki ◽

Sara M. Camp ◽

E. Antonio Chiocca ◽

Xandra O. Breakefield

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Epstein Barr Virus ◽

Single Step ◽

Vector System ◽

Barr Virus ◽

Retrovirus Vector ◽

Vector Sequences ◽

Epstein Barr ◽

Simplex Virus

ABSTRACT We report here on the development and characterization of a novel herpes simplex virus type 1 (HSV-1) amplicon-based vector system which takes advantage of the host range and retention properties of HSV–Epstein-Barr virus (EBV) hybrid amplicons to efficiently convert cells to retrovirus vector producer cells after single-step transduction. The retrovirus genes gag-pol andenv (GPE) and retroviral vector sequences were modified to minimize sequence overlap and cloned into an HSV-EBV hybrid amplicon. Retrovirus expression cassettes were used to generate the HSV-EBV-retrovirus hybrid vectors, HERE and HERA, which code for the ecotropic and the amphotropic envelopes, respectively. Retrovirus vector sequences encoding lacZwere cloned downstream from the GPE expression unit. Transfection of 293T/17 cells with amplicon plasmids yielded retrovirus titers between 106 and 107 transducing units/ml, while infection of the same cells with amplicon vectors generated maximum titers 1 order of magnitude lower. Retrovirus titers were dependent on the extent of transduction by amplicon vectors for the same cell line, but different cell lines displayed varying capacities to produce retrovirus vectors even at the same transduction efficiencies. Infection of human and dog primary gliomas with this system resulted in the production of retrovirus vectors for more than 1 week and the long-term retention and increase in transgene activity over time in these cell populations. Although the efficiency of this system still has to be determined in vivo, many applications are foreseeable for this approach to gene delivery.

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Retroviral expression of recombinant p47phox protein by Epstein-Barr virus-transformed B lymphocytes from a patient with autosomal chronic granulomatous disease

Blood ◽

10.1182/blood.v79.7.1829.bloodjournal7971829 ◽

1992 ◽

Vol 79 (7) ◽

pp. 1829-1835 ◽

Cited By ~ 1

Author(s):

CS Cobbs ◽

HL Malech ◽

TL Leto ◽

SM Freeman ◽

RM Blaese ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Cell Lines ◽

Epstein Barr Virus ◽

Hematopoietic Cell ◽

Granulomatous Disease ◽

Phagocytic Cells ◽

Hl60 Cells ◽

Barr Virus ◽

Epstein Barr ◽

Hematopoietic Cell Lines

Patients with chronic granulomatous disease (CGD) have recurrent infections resulting from a failure of phagocytic cells to produce superoxide. One third of CGD patients have an autosomal gene defect resulting in absence of p47phox protein, a cytoplasmic component critical to superoxide production by phagocytic cells. cDNA encoding p47phox has been cloned and recombinant p47phox (rp47phox) restores superoxide-generating activity to a cell-free assay containing cell membranes and cytosol from p47phox-deficient CGD neutrophils. The goal of the present study was to determine the feasibility of retrovirus mediated expression of rp47phox in the HL60 and U937 human hematopoietic cell lines, and in an Epstein-Barr virus transformed B- lymphocyte cell line (EBV-BCL) derived from a p47phox-deficient CGD patient. Normal EBV-BCL contain p47phox and generate small amounts of superoxide, while this CGD EBV-BCL lacks any detectable p47phox protein. Defective amphotropic retrovirus containing p47phox sequence inserted in the LXSN vector in sense and antisense orientations were used to transduce HL60, U937, and CGD EBV-BCL. p47phox mRNA sequence was detected in cells transduced with either sense or antisense retroviral constructs while rp47phox protein was detected only with the sense construct. The amount of rp47phox protein produced within these cells was greater than the native p47phox present in uninduced HL60 or U937 cells, but substantially less than that present in normal neutrophils, induced HL60 cells, or even normal EBV-BCL. Differentiation of transduced HL60 cells and the associated production of native p47phox in response to dimethyl sulfoxide was not affected. These studies demonstrate that retrovirus constructs can be used to mediate stable expression of rp47phox protein in human hematopoietic cell lines and can restore rp47phox protein within the cytosol of p47phox-deficient EBV-BCL from patients with CGD.

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Synthesis of Novel Polyphenols Consisted of Ferulic and Gallic Acids, and Their Inhibitory Effects on Phorbol Ester-Induced Epstein–Barr Virus Activation and Superoxide Generation

Bioorganic & Medicinal Chemistry ◽

10.1016/s0968-0896(01)00361-3 ◽

2002 ◽

Vol 10 (4) ◽

pp. 1069-1075 ◽

Cited By ~ 10

Author(s):

Eisaku Nomura ◽

Asao Hosoda ◽

Hideko Morishita ◽

Akira Murakami ◽

Koichi Koshimizu ◽

...

Keyword(s):

Phorbol Ester ◽

Epstein Barr Virus ◽

Inhibitory Effects ◽

Superoxide Generation ◽

Barr Virus ◽

Epstein Barr

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Epstein-Barr virus infection, drug resistance and prognosis in Korean T- and NK-cell lymphomas

Pathology International ◽

10.1046/j.1440-1827.2001.01214.x ◽

2001 ◽

Vol 51 (5) ◽

pp. 355-363 ◽

Cited By ~ 21

Author(s):

Chan Kwon Jung ◽

Kyo Young Lee ◽

Yonggoo Kim ◽

Kyungja Han ◽

Sang In Shim ◽

...

Keyword(s):

Drug Resistance ◽

Virus Infection ◽

Epstein Barr Virus ◽

Nk Cell ◽

Epstein Barr Virus Infection ◽

Barr Virus ◽

Epstein Barr ◽

Barr Virus Infection

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Expression of P-glycoprotein and glutathione-S-transferase in recurrent lymphomas: the possible role of Epstein-Barr virus, immunophenotypes, and other predisposing factors.

Journal of Clinical Oncology ◽

10.1200/jco.1993.11.1.109 ◽

1993 ◽

Vol 11 (1) ◽

pp. 109-115 ◽

Cited By ~ 47

Author(s):

A L Cheng ◽

I J Su ◽

Y C Chen ◽

T C Lee ◽

C H Wang

Keyword(s):

Epstein Barr Virus ◽

Significant Prognostic Factor ◽

Glutathione S Transferase ◽

Barr Virus ◽

P Glycoprotein ◽

Tumor Tissues ◽

Epstein Barr ◽

Mdr 1 ◽

Simultaneous Expression

PURPOSE We previously have reported the poor prognoses of recurrent peripheral T-cell lymphoma (PTCL) and Epstein-Barr virus (EBV)-associated PTCL (J Clin Oncol 7:725-731, 1989; Blood 77:799-808, 1991). To study the role that drug resistance plays in this scenario, we conducted a retrospective study of 23 adult patients. PATIENTS AND METHODS All patients had recurrent lymphoma tissue available for immunophenotyping and screening for the existence of EBV DNA in tumor cells by Southern blot analysis and in situ hybridization. Expression of a multidrug resistance P-glycoprotein ([P-gp]mdr-1) and a glutathione redox cycle-related glutathione-S-transferase pi (GST-pi) was determined by an immunohistochemistry method. RESULTS Expression of mdr-1 or GST-pi was found in 11 (48%) and 12 (52%) cases, respectively. Most (11 of 12) of the GST-pi expression occurred simultaneously with mdr-1. Prechemotherapy tumor tissues were available in 11 cases; only two (18.2%) of these cases expressed mdr-1. Four (36%) of 11 cases that expressed mdr-1 (mdr-1(+)) and nine (90%) of 10 cases that did not express mdr-1 (mdr-1(-)) responded to second-line chemotherapy (P < .05). The survival-after-recurrence (SAR) curves significantly favored mdr-1(-) recurrent lymphoma (P < .05). The mdr-1 expression was further correlated with the immunophenotype and EBV association. All six cases of EBV-associated lymphoma (PTCL, five cases; Hodgkin's disease, one case) had significant simultaneous expression of mdr-1 and GST-pi in their recurrent tumor tissues. CONCLUSION (1) mdr-1 expression is a significant prognostic factor in recurrent lymphomas; (2) high expression of mdr-1 is observed in recurrent EBV-associated PTCL; and (3) GST-pi usually expresses simultaneously with mdr-1 in recurrent lymphomas. The role of EBV in the development of mdr-1 and the biologic significance of the simultaneous expression of mdr-1 and GST-pi in recurrent lymphomas are well worth further exploration.

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