Pecina 2013 Abstract MiP2013

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Pecina P, Houšťková H, Mráček T, Pecinová A, Nůsková H, Tesařová M, Hansíková H, Janota J, Zeman J, Houštěk J (2013) The use of lymphocytes for diagnostics of mitochondrial oxidative phosphorylation disorders. Mitochondr Physiol Network 18.08.
MiP2013, Book of Abstracts Open Access

Pecina P, Houstkova H, Mracek T, Pecinova A, Nuskova H, Tesarova M, Hansikova H, Janota J, Zeman J, Houstek J (2013)

Event: MiPNet18.08_MiP2013

Mitochondrial diseases belong to most severe inherited metabolic diseases affecting the pediatric population. Diagnostics of a substantial part of mitochondrial diseases with unknown genetic cause relies on clinical symptoms and biochemical analysis of energetic function and content of individual mitochondrial proteins in patient tissues – mainly in bioptic samples of skeletal muscle and cell cultures of skin fibroblasts. However, due to their invasive nature, the biopsies are often refused by the parents of the patient. Therefore, we tested the diagnostic suitability of easily obtainable patient material – lymphocytes isolated from peripheral blood.

High-resolution respirometry enables sensitive analysis of the mitochondrial oxidative phosphorylation system in isolated lymphocytes. Substrate-inhibitor measurements in digitonin-permeabilized lymphocytes provide a complex evaluation of individual respiratory complexes, coupled ATP synthesis and kinetic parameters of mitochondrial respiratory enzymes. We employed this approach in a large cohort of 48 children including mostly subjects with suspected mitochondrial disease, previously diagnosed patients with OXPHOS disorders, and controls. In combination with cytofluorometric detection of mitochondrial membrane potential and protein analysis by SDS and native electrophoreses it was possible to diagnose specific defects of Complexes I, CIV and CV using small amounts of peripheral blood within 1-2 days. Importantly, functional manifestations of mitochondrial disorders caused by SURF1 [1] and TMEM70 [2] mutations in lymphocytes recapitulate previous findings in fibroblasts.

The noninvasiveness, reliability and speed of such an approach demonstrate the high potential of isolated lymphocytes for diagnostics of oxidative phosphorylation disorders in patients with suspected mitochondrial disease.

MiPNetLab: CZ Prague Houstek J, CZ Prague Zeman J

Labels: MiParea: Respiration, mtDNA;mt-genetics, mt-Medicine, Patients Mammal;model: Human Tissue;cell: Blood cells  Cell line: Lymphocyte Preparation: Permeabilized cells, Homogenate Enzyme: Complex I, Complex IV;cytochrome c oxidase, Complex V;ATP synthase Pathology: Inherited Regulation: mt-Membrane potential Coupling state: LEAK, OXPHOS, ETS Substrate state: CI, CII, CGpDH, CI&II, Other combinations, ROX HRR: Oxygraph-2k Additional: MiP2013 

Affiliations, acknowledgements and author contributions

  1. Dept of Bioenergetics, Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic;
  2. Dept of Pediatrics and Adolescent Medicine;
  3. Dept of Neonatology, Thomayer Hospital, Prague, Czech Republic;
  4. Clinic of Pediatrics and Adolescent Medicine, General University Hospital, Prague, Czech Republic.



  1. Pecina P, Houstková H, Hansíková H, Zeman J, Houstek J (2004) Genetic defects of cytochrome c oxidase assembly. Physiol Res 53 Suppl 1: 213-123.
  2. Cizkova A, Stranecky V, Mayr JA, Tesarova M, Havlickova V, Paul J, Ivanek R, Kuss AW, Hansikova H, Kaplanova V, Vrbacky M, Hartmannova H, Noskova L, Honzik T, Drahota Z, Magner M, Hejzlarova K, Sperl W, Zeman J, Houstek J, Kmoch S (2008) TMEM70 mutations cause isolated ATP synthase deficiency and neonatal mitochondrial encephalocardiomyopathy. Nat Genet 40: 1288-1290.


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