Lymphocyte-based determination of susceptibility to malignant hyperthermia: a pilot study in swine.

Abstract

BACKGROUND:

Malignant hyperthermia susceptibility (MHS) is diagnosed by an invasive in vitro caffeine-halothane contracture test (CHCT) carried out on biopsied skeletal muscle tissue. We are presenting a novel blood test approach for malignant hyperthermia testing in a swine model. Our main aim was to determine whether adenosine production from lymphocytes after 4-chloro-m-cresol (4CmC) stimulation distinguishes homozygous swine carrying the Arg615Cys mutation in the ryanodine receptor type 1 (RyR1) gene (MHS swine) from normal swine.

METHODS:

Lymphocytes were isolated from arterial blood (40 ml) obtained from MHS (n = 7) and normal (n = 7) swine. Cells were suspended in Hank's balanced salt solution and treated with 4CmC (0-10 mm) at 37°C in the presence of adenosine deaminase inhibitor. After termination and purification of samples, aliquots (50 μl) were assayed for adenosine content using high performance liquid chromatography.

RESULTS:

Baseline adenosine levels before stimulating lymphocytes with 4CmC were 0.025 ± 0.004 and 0.041 ± 0.006 μm (mean ± SEM) in lymphocytes from normal and MHS swine, respectively (P = 0.125). Maximum responses were achieved at 1 mm 4CmC for both cell-line groups. Adenosine levels after stimulation with 4CmC (1 mm) were 0.185 ± 0.009 and 0.397 ± 0.038 μm in lymphocytes from normal and MHS swine, respectively (P = 0.0035). There was no overlap between adenosine levels in stimulated lymphocytes from MHS and normal swine.

CONCLUSION:

4CmC stimulation of porcine lymphocytes induces increased adenosine formation in MHS cells relative to those from normal swine; evaluation of adenosine formation in response to RyR1 agonists in human lymphocytes is needed.

• Authors: Saiid Bina, John Capacchione, Sheila Muldoon, Munkhuu Bayarsaikhan, Rolf Bunger
• Journal: Anesthesiology (Anesthesiology) Vol. 113 Issue 4 Pg. 917-24 (Oct 2010) ISSN: 1528-1175 [Electronic] United States
• PMID: 20823761 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Adenosine Deaminase Inhibitors
• Enzyme Inhibitors
• Imidazoles
• Muscle Relaxants, Central
• Oxazoles
• Ryanodine Receptor Calcium Release Channel
• Uncoupling Agents
• Inosine
• azumolene
• Adenosine Triphosphate
• Magnesium
• Adenosine
• Calcium

Topics

• Adenosine (metabolism)
• Adenosine Deaminase Inhibitors
• Adenosine Triphosphate (metabolism)
• Animals
• Calcium (metabolism, pharmacology)
• Chromatography, High Pressure Liquid
• Disease Susceptibility
• Enzyme Inhibitors (pharmacology)
• Imidazoles (pharmacology)
• Inosine (metabolism)
• Lymphocytes (physiology)
• Magnesium (pharmacology)
• Malignant Hyperthermia (diagnosis)
• Muscle Relaxants, Central (pharmacology)
• Oxazoles (pharmacology)
• Oxidative Phosphorylation (drug effects)
• Pilot Projects
• Ryanodine Receptor Calcium Release Channel (physiology)
• Sarcoplasmic Reticulum (drug effects)
• Swine
• Uncoupling Agents (pharmacology)