Both granulocytic and non-granulocytic blood cells are affected in patients with severe congenital neutropenia and their non-neutropenic family members: An evaluation of morphology, function and cell death

Abstract

Objective:To examine granulocytic and non-granulocytic cells in children with severe congenital neutropenia (SCN) and their non-neutropenic parents.Materials and Methods:Fifteen patients with SCN and 21 non-neutropenic parents were evaluated for a) CD95, CD95 ligand, annexin V, propidium iodide, cell cycle, and lymphocyte subsets by flow cytometry; b) rapid cell senescence (of leukocytes) by senescence-associated β-galactosidase stain; c) aggregation tests by aggregometer; d) in vitro bleeding time by PFA-100 instrument; e) mepacrine-labeled dense granule number of thrombocytes by fluorescence microscope; and f) hematomorphology by light and electron microscope. HAX1, ELANE, G6PC3, CSF3R, and JAGN1 mutations associated with SCN were studied in patients and several parents.Results:Significant increase in apoptosis and secondary necrosis in monocytes, lymphocytes, and granulocytes of the patients and parents was detected, irrespective of the mutation type. CD95 and CD95 ligand results implied that apoptosis was non-CD95-mediated. Leukocytes of 25%, 12.5%, and 0% of patients, parents, and controls showed rapid cell senescence. The cell cycle analysis testable in four cases showed G1 arrest and apoptosis in lymphocytes of three. The patients had HAX1 (n=6), ELANE (n=2), G6PC3 (n=2), and unidentified (n=5) mutations. The CD3, CD4, and NK lymphocytes were below normal levels in 16.6%, 8.3%, and 36.4% of the patients and in 0%, 0%, and 15.4% of the parents (controls: 0%, 0%, 5.6%). The thrombocytes aggregated at low rates, dense granule number/thrombocyte ratio was low, and in vitro bleeding time was prolonged in 37.5%-66.6% of patients and 33.3%-63.2% of parents (vs. 0% in controls). Under electron and/or light microscope, the neutrophils, monocytes, lymphocytes, and thrombocytes in the peripheral blood of both patients and parents were dysplastic and the bone marrow of patients revealed increased phagocytic activity, dysmegakaryopoiesis, and necrotic and apoptotic cells. Ultrastructurally, thrombocyte adhesion, aggregation, and release were inadequate.Conclusion:In cases of SCN, patients’ pluripotent hematopoietic stem cells and their non-neutropenic parents are both affected irrespective of the genetic defect.

Amyloidosis in a Patient With Congenital Neutropenia Because of G6PC3 Deficiency

“…Most of the mutations in this gene are on exon 1 (mostly nonsense and splice site mutations) and exon 6 (mostly missense) regions 1. The G6PC3 variant found in our patient and in his sister was on exon 1 p.E65A (c.194A>C) region 8. A new nonsyndromic mutation was defined on Exon 3 9.…”

“…During follow-up, as a part of a national collaborative project, the siblings were re-tested for congenital neutropenia syndromes. Homozygous mutation p.E65A (c.194A>C) of the G6PC3 gene was identified in both patients 8…”

“…1 The G6PC3 variant found in our patient and in his sister was on exon 1 p.E65A (c.194A > C) region. 8 A new nonsyndromic mutation was defined on Exon 3. 9 Only 6 patients with nonsyndromic mutations were reported in the literature.…”

Novel ELANE Gene Mutation in a Newborn with Severe Congenital Neutropenia: Case Report and Literature Review

“…[21][22][23][24][25] Some researchers found that both granulocytic and nongranulocytic blood cells were affected in SCN patients. 26 ELANE gene mutation-related SCN usually presents with neutropenia and no other extra-hematopoietic manifestations, 27 unlike other gene mutations associated with SCN. However, known and novel ELANE gene mutations and variations are important for the homeostasis and functioning not only of the neutrophil granulocyte but also of the entire hematopoietic system.…”

“Chewing Technique” Using Gums toward Mucositis Prevalence on Chemotherapeutic Cancer Patients

“…Currently, there is no concensus regarding proper treatment in oral care to prevent mucositis due to chemotherapy. 5 Until recently, no consent has been done regarding the proper treatment in oral care to prevent mucositis due to chemotherapy. Although several implementations with therapy intervention are being developed, no effective action has come to fruition regarding oral care treatment to prevent mucositis due to chemotherapy.…”