Somatic Triple Mosaicism in a Carrier of X-Linked Chronic Granulomatous Disease

Abstract The X-linked form of chronic granulomatous disease (CGD) is caused by mutations in the CYBB gene, which encodes the 91-kD subunit of the flavocytochrome b558, a component of the superoxide-generating nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in phagocytic leukocytes. Mutations in this gene are very heterogeneous and often unique for one family. Here we report on a family with two patients (brothers), one with a 3-kb deletion comprising exon 5 and the other with a 3.5-kb deletion comprising exons 6 and 7 of the CYBB gene. Sequence analysis of polymerase chain reaction (PCR)-amplified genomic DNA proved these deletions to be overlapping for 35 bp. Analysis by restriction fragment length polymorphism of genomic DNA from the mother's leukocytes showed her to be a carrier of both deletions in addition to the normal CYBB sequence. This triple somatic mosaicism was confirmed with PCR-amplified genomic and complementary DNA. The presence of the normal CYBB gene in the mother was also proven by the finding of normal superoxide-generating neutrophils in addition to cells lacking this ability. Triple X syndrome was excluded. These findings suggest that the mutations are the result of an event in early embryogenesis of the mother, possibly involving a mechanism like sister chromatid exchange.

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Somatic Triple Mosaicism in a Carrier of X-Linked Chronic Granulomatous Disease

Blood ◽

10.1182/blood.v91.1.252.252_252_257 ◽

1998 ◽

Vol 91 (1) ◽

pp. 252-257

Author(s):

Martin de Boer ◽

Egbert Bakker ◽

Stefaan Van Lierde ◽

Dirk Roos

Keyword(s):

Chronic Granulomatous Disease ◽

Genomic Dna ◽

Gene Sequence ◽

Somatic Mosaicism ◽

Nicotinamide Adenine Dinucleotide Phosphate ◽

Granulomatous Disease ◽

Gene Sequence Analysis ◽

Triple X Syndrome ◽

Cybb Gene ◽

Polymerase Chain

The X-linked form of chronic granulomatous disease (CGD) is caused by mutations in the CYBB gene, which encodes the 91-kD subunit of the flavocytochrome b558, a component of the superoxide-generating nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in phagocytic leukocytes. Mutations in this gene are very heterogeneous and often unique for one family. Here we report on a family with two patients (brothers), one with a 3-kb deletion comprising exon 5 and the other with a 3.5-kb deletion comprising exons 6 and 7 of the CYBB gene. Sequence analysis of polymerase chain reaction (PCR)-amplified genomic DNA proved these deletions to be overlapping for 35 bp. Analysis by restriction fragment length polymorphism of genomic DNA from the mother's leukocytes showed her to be a carrier of both deletions in addition to the normal CYBB sequence. This triple somatic mosaicism was confirmed with PCR-amplified genomic and complementary DNA. The presence of the normal CYBB gene in the mother was also proven by the finding of normal superoxide-generating neutrophils in addition to cells lacking this ability. Triple X syndrome was excluded. These findings suggest that the mutations are the result of an event in early embryogenesis of the mother, possibly involving a mechanism like sister chromatid exchange.

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Chronic Granulomatous Disease Presenting as Aseptic Ascites in a 2-Year-Old Child

Case Reports in Immunology ◽

10.1155/2013/927897 ◽

2013 ◽

Vol 2013 ◽

pp. 1-5 ◽

Cited By ~ 1

Author(s):

J. F. Moreau ◽

John A. Ozolek ◽

P. Ling Lin ◽

Todd D. Green ◽

Elaine A. Cassidy ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Fungal Infections ◽

Novel Mutation ◽

Nicotinamide Adenine Dinucleotide Phosphate ◽

Unknown Origin ◽

Granulomatous Disease ◽

Cybb Gene ◽

Inherited Immunodeficiency ◽

Peritoneal Biopsy ◽

Immunodeficiency Syndrome

Chronic granulomatous disease (CGD) is a rare inherited immunodeficiency syndrome that results from abnormal nicotinamide adenine dinucleotide phosphate (NADPH) oxidase function. This defect leads to recurrent catalase-positive bacterial and fungal infections as well as associated granuloma formation. We review the case of a 2-year-old boy who presented with ascites and fever of an unknown origin as manifestations of CGD. Cultures were negative for infection throughout his course, and CGD was suspected after identification of granulomas on peritoneal biopsy. Genetic testing revealed a novel mutation in the CYBB gene underlying his condition. This paper highlights the importance of considering CGD in the differential diagnosis of fever of unknown origin and ascites in children.

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A new mutation in exon 12 of the gp91-phox gene leading to cytochrome b- positive X-linked chronic granulomatous disease

Blood ◽

10.1182/blood.v85.11.3274.bloodjournal85113274 ◽

1995 ◽

Vol 85 (11) ◽

pp. 3274-3277 ◽

Cited By ~ 14

Author(s):

H Azuma ◽

H Oomi ◽

K Sasaki ◽

I Kawabata ◽

T Sakaino ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Cytochrome B ◽

Dna Analysis ◽

Nicotinamide Adenine Dinucleotide Phosphate ◽

Granulomatous Disease ◽

New Mutation ◽

Nadph Oxidase Complex ◽

Polymerase Chain ◽

O2 Production ◽

Complete Formation

We have previously reported a patient with cytochrome b-positive X-linked chronic granulomatous disease. Although the O2-production of neutrophils from the patient was completely defective, we presented data suggesting that the patient's cytochrome b was present at a normal level and possibly had normal spectroscopic features. Thus, to look for a mutation in the cytochrome b heavy chain (gp91-phox) gene, DNA analysis of gp91-phox cDNA derived from this patient was performed. As a result, we found that five nucleotides (1521 through 1525) within exon 12 were deleted, and a new sequence of eight nucleotides was inserted. This mutation converted Gln507-Lys508-Thr509 into His-Ile-Trp-Ala. Mismatched polymerase chain reaction showed that the mother has both wild and mutated alleles, confirming that this case was transmitted in an X-linked fashion. This mutation is different from those previously reported by others. The translocation of p47-phox and p67-phox to the membrane fraction occurred, indicating the complete formation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. We conclude that this case suggests that the structure encoded on exon 12 of gp91-phox is important for electron transfer.

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Systemic Redox Imbalance in Patients with Chronic Granulomatous Disease

Journal of Clinical Medicine ◽

10.3390/jcm9051397 ◽

2020 ◽

Vol 9 (5) ◽

pp. 1397 ◽

Cited By ~ 1

Author(s):

Edyta Heropolitanska-Pliszka ◽

Klaudia Berk ◽

Mateusz Maciejczyk ◽

Jolanta Sawicka-Powierza ◽

Ewa Bernatowska ◽

...

Keyword(s):

Oxidative Damage ◽

Chronic Granulomatous Disease ◽

Coenzyme Q10 ◽

Nicotinamide Adenine Dinucleotide Phosphate ◽

Redox Status ◽

Oxidation Products ◽

Stress Index ◽

Granulomatous Disease ◽

Redox Imbalance ◽

Cybb Gene

The aim of our study was to evaluate redox status, enzymatic and non-enzymatic antioxidant barriers, oxidative damage of proteins, lipids and DNA, as well as concentration of coenzyme Q10 and vitamins A and E in patients with chronic granulomatous disease (CGD). The study was performed on fifteen Caucasian individuals (median age 24 years and seven months) diagnosed with CGD. The mutation in the NCF1 gene was confirmed in ten patients, and in the CYBB gene in five patients. We demonstrated high levels of total oxidant status (TOS) and oxidative stress index (OSI), lipids (↑8-isoprostanes (8-isoP), ↑4-hydroxynonenal (4-HNE)), proteins (↑advanced oxidation protein products (AOPP)) and DNA (↑8-hydroxy-2’-deoxyguanosine (8-OHdG)) oxidation products in CGD individuals as compared to sex- and age-matched healthy controls. We showed enhanced serum enzymatic activity of catalase (CAT) and superoxide dismutase-1 (SOD) and significantly decreased coenzyme Q10 concentration. Our study confirmed redox disturbances and increased oxidative damage in CGD patients, and indicated the need to compare redox imbalance depending on the type of mutation and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity. The question regarding effectiveness of antioxidant therapy in patients with CGD is open, and the need to establish guidelines in this area remains to be addressed.

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Genetic analysis of CYBB gene in 26 korean families with X-linked chronic granulomatous disease

Immunological Investigations ◽

10.3109/08820139.2013.825270 ◽

2014 ◽

Vol 43 (6) ◽

pp. 585-594 ◽

Cited By ~ 1

Author(s):

Sun Hi Ko ◽

Jung Woo Rhim ◽

Kyung Sue Shin ◽

Youn Soo Hahn ◽

So Young Lee ◽

...

Keyword(s):

Genetic Analysis ◽

Chronic Granulomatous Disease ◽

Granulomatous Disease ◽

Korean Families ◽

Cybb Gene

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Identification of a Novel Mutation in the CYBB Gene, p.Asp378Gly, in a Patient With X-linked Chronic Granulomatous Disease

Allergy Asthma and Immunology Research ◽

10.4168/aair.2014.6.4.366 ◽

2014 ◽

Vol 6 (4) ◽

pp. 366 ◽

Cited By ~ 1

Author(s):

Sang-Mi Song ◽

Mi-Ran Park ◽

Do-Soo Kim ◽

Jihyun Kim ◽

Yae-Jean Kim ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Novel Mutation ◽

Granulomatous Disease ◽

Cybb Gene

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X-Linked Chronic Granulomatous Disease: Mutations in the CYBB Gene Encoding the gp91-phox Component of Respiratory-Burst Oxidase

The American Journal of Human Genetics ◽

10.1086/301874 ◽

1998 ◽

Vol 62 (6) ◽

pp. 1320-1331 ◽

Cited By ~ 121

Author(s):

Julie Rae ◽

Peter E. Newburger ◽

Mary C. Dinauer ◽

Deborah Noack ◽

Penelope J. Hopkins ◽

...

Keyword(s):

Chronic Granulomatous Disease ◽

Respiratory Burst ◽

Granulomatous Disease ◽

Gene Encoding ◽

Disease Mutations ◽

Cybb Gene ◽

Respiratory Burst Oxidase

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Interferon-gamma improves splicing efficiency of CYBB gene transcripts in an interferon-responsive variant of chronic granulomatous disease due to a splice site consensus region mutation

Blood ◽

10.1182/blood.v95.11.3548 ◽

2000 ◽

Vol 95 (11) ◽

pp. 3548-3554 ◽

Cited By ~ 51

Author(s):

Antonio Condino-Neto ◽

Peter E. Newburger

Keyword(s):

B Cell ◽

Chronic Granulomatous Disease ◽

Cell Line ◽

Interferon Gamma ◽

Granulomatous Disease ◽

First Intron ◽

Cybb Gene ◽

Nuclear Rna ◽

Ifn Γ

Abstract X-linked chronic granulomatous disease (CGD) derives from defects in the CYBB gene, which encodes the gp91-phox component of NADPH oxidase. We studied the molecular basis of the disease in a kindred with variant CGD, due to a single base substitution at the sixth position of CYBB first intron. The patients' phagocytes have been shown previously to greatly increase superoxide release in response to interferon-gamma (IFN-γ) in vitro and in vivo. We examined CYBB gene expression in an Epstein-Barr virus (EBV)-transformed B-cell line from 1 patient in this kindred. These cells showed markedly decreased levels of CYBB transcripts in total RNA (5% of normal) and nuclear RNA (1.4% of normal), despite equal CYBB transcription rates in the CGD and control cells. Incubation with IFN-γ produced a 3-fold increase in CYBBtotal messenger RNA (mRNA) levels in the patient's cells, and decreased nuclear transcripts to undetectable levels. Reverse transcriptase–polymerase chain reaction analysis of RNA splicing revealed a preponderance of unspliced CYBB transcripts in the patient's nuclear RNA. In vitro incubation with IFN-γ increased by 40% the ratio of spliced relative to unspliced CYBB mRNA in nuclei from the CGD B-cell line. Total RNA harvested from the same patient's monocytes, on and off therapy with IFN-γ, showed a similar improvement in splicing. We conclude that IFN-γ partially corrects a nuclear processing defect due to the intronic mutation in theCYBB gene in this kindred, most likely by augmentation of nuclear export of normal transcripts, and improvement in the fidelity of splicing at the first intron.

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NADPH oxidase correction by mRNA transfection of apheresis granulocytes in chronic granulomatous disease

Blood Advances ◽

10.1182/bloodadvances.2020003224 ◽

2020 ◽

Vol 4 (23) ◽

pp. 5976-5987

Author(s):

Suk See De Ravin ◽

Julie Brault ◽

Ronald J. Meis ◽

Linhong Li ◽

Narda Theobald ◽

...

Keyword(s):

Nadph Oxidase ◽

Chronic Granulomatous Disease ◽

Messenger Rna ◽

Nicotinamide Adenine Dinucleotide Phosphate ◽

Functional Restoration ◽

In Vivo Studies ◽

Allogeneic Bone ◽

Granulomatous Disease ◽

Primate Model

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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