Mechanisms of Microtubule Disassembly in Vivo: Studies in Normal and Chronic Granulomatous Disease Leucocytes

Abstract Granulocytes from patients with chronic granulomatous disease (CGD) have dysfunctional phagocyte reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase that fails to generate sufficient antimicrobial reactive oxidative species. CGD patients with severe persistent fungal or bacterial infection who do not respond to antibiotic therapy may be given apheresis-derived allogeneic granulocyte transfusions from healthy volunteers to improve clearance of intractable infections. Allogeneic granulocyte donors are not HLA matched, so patients who receive the donor granulocyte products may develop anti-HLA alloimmunity. This not only precludes future use of allogeneic granulocytes in an alloimmunized CGD recipient, but increases the risk of graft failure of those recipients who go on to need an allogeneic bone marrow transplant. Here, we provide the first demonstration of efficient functional restoration of CGD patient apheresis granulocytes by messenger RNA (mRNA) electroporation using a scalable, Good Manufacturing Practice–compliant system to restore protein expression and NADPH oxidase function. Dose-escalating clinical-scale in vivo studies in a nonhuman primate model verify the feasibility, safety, and persistence in peripheral blood of infusions of mRNA-transfected autologous granulocyte-enriched apheresis cells, supporting this novel therapeutic approach as a potential nonalloimmunizing adjunct treatment of intractable infections in CGD patients.

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176. In Vivo Selective Enhancement of Correction of Human X-Linked Chronic Granulomatous Disease Neutrophils in NOD/SCID Mice Transplanted with CD34+ Stem Cells Transduced with a Bicistronic gp91phox/Benzyl Guanine Resistant Methylguanine Methyltransferase Vector

Molecular Therapy ◽

10.1016/j.ymthe.2004.06.136 ◽

2004 ◽

Vol 9 ◽

pp. S67

Keyword(s):

Stem Cells ◽

Chronic Granulomatous Disease ◽

Scid Mice ◽

Granulomatous Disease ◽

Methylguanine Methyltransferase ◽

Selective Enhancement

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Improvement of superoxide production in monocytes from patients with chronic granulomatous disease by recombinant cytokines

Blood ◽

10.1182/blood.v81.8.2131.bloodjournal8182131 ◽

1993 ◽

Vol 81 (8) ◽

pp. 2131-2136

Author(s):

V Jendrossek ◽

AM Peters ◽

S Buth ◽

J Liese ◽

U Wintergerst ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Respiratory Burst ◽

Interleukin 1 ◽

Superoxide Production ◽

Granulomatous Disease ◽

Necrosis Factor Alpha ◽

Ifn Gamma ◽

Phorbolmyristate Acetate

Cytokines have been shown to modulate the respiratory burst of polymorphonuclear leukocytes and monocytes from normal controls. We have examined whether monocytes from children with chronic granulomatous disease (CGD) can be primed by cytokines other than interferon-gamma (IFN gamma), which has been demonstrated to improve the production of reactive oxygen species in vivo and in vitro. Monocytes isolated from peripheral blood were cultured without and with IFN gamma (500 U/mL), tumor necrosis factor-alpha (500 U/mL), interleukin-1 beta (IL-1 beta) (100 U/mL), and IL-3 (100 U/mL). After 3 days of culture, the phorbolmyristate acetate (2 ng/mL) and the formyl- methionyl-leucyl-phenylalanine (0.1 mumol/L)-stimulated superoxide- production was determined in a microtiter system. In nearly all of the 14 patients examined (5 autosomal, 5 X-chromosomal, and 4 of unknown inheritance), an improvement of superoxide production could be demonstrated. The most impressive effect with the cytokines newly tested was seen with monocytes from autosomal CGD patients treated with IL-3 and stimulated by phorbolmyristate acetate. In single patients cultivation of monocytes with IL-6 and granulocyte-macrophage colony- stimulating factor resulted in only slight improvement of superoxide production. Our findings indicate that cytokines other than IFN gamma can positively modulate the defective respiratory burst in CGD and that each patient reacts with an individual pattern to different cytokines.

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PEG-fDAO Reduces Lung Inflammation in Chronic Granulomatous Disease Mice Post Challenge with Nonviable Candida Albicans

10.21203/rs.3.rs-216646/v1 ◽

2021 ◽

Author(s):

Hiroyuki Nunoi ◽

Peiyu Xie ◽

Hideaki Nakamura ◽

Yasuaki Aratani ◽

Jun Fang ◽

...

Keyword(s):

Candida Albicans ◽

Amino Acid ◽

Chronic Granulomatous Disease ◽

Lung Inflammation ◽

Acid Oxidase ◽

Granulomatous Disease ◽

Amino Acid Oxidase ◽

Lung Weight ◽

Inflammation Model

Abstract We previously reported that polyethylene glycol-conjugated recombinant porcine D-amino acid oxidase (PEG-pDAO) could supply reactive oxygen species (ROS) to defective NADPH oxidase in neutrophils of patients with chronic granulomatous disease (CGD), and neutrophils regain bactericidal activity in vitro. In the present study, we employed an in vivo nonviable Candida albicans (nCA)-induced lung inflammation model using gp91-phox knockout CGD mice and novel PEG conjugates of Fusarium spp. D-amino acid oxidase (PEG-fDAO), rather than PEG-pDAO. Using three experimentation strategies with the in vivo lung inflammation model, the mouse body weight, lung weight, and lung pathology were evaluated to confirm the efficacy of ROS-generating enzyme replacement therapy with PEG-fDAO. The lung weight and pathological findings were significantly ameliorated by the administration of PEG-fDAO followed by intraperitoneal injection of D-phenylalanine or D-proline. These data suggest that PEG- fDAO with the function of targeted delivery to the nCA-induced inflammation site is applicable in the treatment of inflammation in CGD in vivo.

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Reconstitution of defective respiratory burst activity with partially purified human neutrophil cytochrome B in two genetic forms of chronic granulomatous disease: possible role of Rap1A

Blood ◽

10.1182/blood.v79.9.2438.2438 ◽

1992 ◽

Vol 79 (9) ◽

pp. 2438-2445 ◽

Cited By ~ 9

Author(s):

MT Quinn ◽

JT Curnutte ◽

CA Parkos ◽

ML Mullen ◽

PJ Scott ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Cytochrome B ◽

Human Neutrophil ◽

Autosomal Recessive ◽

Plasma Membranes ◽

Close Association ◽

Granulomatous Disease ◽

Detergent System

Abstract Neutrophil plasma membranes from patients with the X-linked and autosomal recessive forms of chronic granulomatous disease (CGD) that lack cytochrome b are incapable of generating superoxide anion (O2-) in vivo and in vitro. The O2- generating activity of these defective membranes was reconstituted with the addition of partially purified human neutrophil cytochrome b in a detergent-based, cell-free activation system. Depending on the detergent system used, 50% to 100% of the activity of control membranes was recovered, and this activity was directly dependent on the cytochrome b concentration. However, when cytochrome b was purified to 99% homogeneity, the reconstitutive capacity of the cytochrome was lost, possibly because of subtle denaturation of the cytochrome or the removal of an additional required cofactor. Examination of the latter possibility with respect to a protein known to coassociate with the cytochrome, ie, Rap1A, indicated that this ras-like protein was present in the partially purified cytochrome preparation used to reconstitute activity in CGD membranes, but was missing in the highly purified preparation. However, the finding that Rap1A was present in normal amounts in the neutrophil membranes from all four major types of CGD (including those missing cytochrome b) suggested that the conditions required of the reconstitution assay did not favor the reassociation of the membrane- derived Rap1A with exogenously added cytochrome b or that another unidentified membrane component was lost during the final purification step. The normal expression of Rap1A in CGD cell membranes also indicates that this protein is not responsible for the absence of O2- production in the X-linked and autosomal recessive cytochrome b- negative forms of CGD. Finally, these results show that the expression of Rap1A in the plasma membrane is not dependent on the coordinate expression of cytochrome b, despite the close association shown for these two proteins in the normal cell membrane.

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Restoration of phagocyte function by interferon-gamma in X-linked chronic granulomatous disease occurs at the level of a progenitor cell

Blood ◽

10.1182/blood.v76.12.2443.2443 ◽

1990 ◽

Vol 76 (12) ◽

pp. 2443-2448 ◽

Cited By ~ 43

Author(s):

RA Ezekowitz ◽

CA Sieff ◽

MC Dinauer ◽

DG Nathan ◽

SH Orkin ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Progenitor Cells ◽

Interferon Gamma ◽

Granulomatous Disease ◽

Daughter Cells ◽

Myeloid Progenitor ◽

Myeloid Progenitor Cells ◽

Nbt Reduction ◽

Rare Cells

Abstract Phagocytes from X-linked chronic granulomatous disease (X-CGD) patients are deficient in their ability to generate superoxide because of a defective gene that encodes a heavy chain of cytochrome b, a critical component in the superoxide-generating pathway. Previously we have shown that a single in vivo treatment of selected X-CGD patients with interferon-gamma (INF-gamma) resulted 14 days later in near-normal levels of superoxide generation by phagocytes. The effect persisted for 28 days. This prolonged effect suggested that the lymphokine affected progenitor cells. In this study, we examined progenitor-derived colonies from the peripheral blood from this unusual X-CGD kindred. Progenitor-derived colonies examined before treatment were unable to generate superoxide as visualized by lack of nitro blue tetrazolium (NBT) reduction compared with normal controls. By contrast, colonies derived 7 days after a single INF-gamma injection were able to generate superoxide as shown by increased NBT reduction. Colonies harvested 21 days after treatment contained only rare cells capable of NBT reduction. Our results indicate that INF-gamma can reprogram the myeloid progenitor cells to express a partially corrected phenotype. This corrected phenotype is later expressed in daughter cells.

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723. In Vivo Expansion of MDS1/EVI1, PRDM16 and SETBP1 Integration Clones in Successful Chronic Granulomatous Disease (CGD) Gene Therapy Trial

Molecular Therapy ◽

10.1016/j.ymthe.2006.08.803 ◽

2006 ◽

Vol 13 ◽

pp. S279

Author(s):

Kerstin Schwarzwaelder ◽

Manfred Schmidt ◽

Marion G. Ott ◽

Stefan Stein ◽

Hanno Glimm ◽

...

Keyword(s):

Gene Therapy ◽

Chronic Granulomatous Disease ◽

Granulomatous Disease ◽

Gene Therapy Trial ◽

Therapy Trial

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Different Degrees of NADPH Oxidase 2 Regulation and In Vivo Platelet Activation: Lesson From Chronic Granulomatous Disease

Journal of the American Heart Association ◽

10.1161/jaha.114.000920 ◽

2014 ◽

Vol 3 (3) ◽

Cited By ~ 22

Author(s):

Roberto Carnevale ◽

Lorenzo Loffredo ◽

Valerio Sanguigni ◽

Alessandro Plebani ◽

Paolo Rossi ◽

...

Keyword(s):

Nadph Oxidase ◽

Chronic Granulomatous Disease ◽

Platelet Activation ◽

Granulomatous Disease ◽

Nadph Oxidase 2

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In Vivo Interferon- Therapy Augments the In Vitro Ability of Chronic Granulomatous Disease Neutrophils to Damage Aspergillus Hyphae

The Journal of Infectious Diseases ◽

10.1093/infdis/163.4.849 ◽

1991 ◽

Vol 163 (4) ◽

pp. 849-852 ◽

Cited By ~ 77

Author(s):

J. H. Rex ◽

J. E. Bennett ◽

J. I. Gallin ◽

H. L. Malech ◽

E. S. DeCarlo ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Interferon Therapy ◽

Granulomatous Disease

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Enhanced Host Defense After Gene Transfer in the Murine p47phox-Deficient Model of Chronic Granulomatous Disease

Blood ◽

10.1182/blood.v89.7.2268 ◽

1997 ◽

Vol 89 (7) ◽

pp. 2268-2275 ◽

Cited By ~ 101

Author(s):

Michael Mardiney ◽

Sharon H. Jackson ◽

S. Kaye Spratt ◽

Fei Li ◽

Steven M. Holland ◽

...

Keyword(s):

Gene Therapy ◽

Gene Transfer ◽

Chronic Granulomatous Disease ◽

Host Defense ◽

Ex Vivo ◽

Pseudomonas Cepacia ◽

Granulomatous Disease ◽

Human Immune System ◽

Human Form

Abstract The p47phox−/− mouse exhibits a phenotype similar to that of human chronic granulomatous disease (CGD) and, thus, is an excellent model for the study of gene transfer technology. Using the Moloney murine leukemia virus–based retroviral vector MFG-S encoding the human form of p47phox, we performed ex vivo gene transfer into Sca-1+ p47phox−/− marrow progenitor cells without conditioning of donors with 5-fluorouracil. Transduced progenitors were transplanted into moderately irradiated (500 cGy), G-CSF preconditioned sibling p47phox−/− mice. Using the fluorescent probe dihydrorhodamine 123 (DHR), in vivo biochemical correction of the superoxide-generating NADPH oxidase system was detected by flow cytometry in 12.3% ± 0.9% of phorbol myristate acetate–stimulated peripheral blood neutrophils at 4 weeks and 2.6% ± 1.0% at 14 weeks after transplantation. Following gene therapy, mice were challenged with the CGD pathogen Burkholderia (formerly Pseudomonas) cepacia and bacteremia levels were assessed at 24 hours and 7 days after inoculation. At both time points, bacteremia levels in gene corrected p47phox−/− mice were significantly lower than untreated p47phox−/− mice (0.89 ± 0.30 colonies v 237.7 ± 83.6 colonies at 24 hours, P < .02; 4.0 ± 2.0 colonies v 110.2 ± 26.5 colonies at 7 days, P < .0014). More importantly, Kaplan-Meier survival analysis showed a significant survival advantage of gene corrected versus untreated p47phox−/− mice (P < .001). Thus, stem-cell–directed ex vivo gene therapy is capable of restoring phagocyte oxidant-dependent host-defense function in this mouse model of a human immune-system disorder.

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