FACTORS INFLUENCING HUMAN POLYMORPHONUCLEAR LEUKOCYTE CHEMILUMINESCENCE

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Abstract
Following particle ingestion, the microbicidal mechanisms of polymorphonuclear leukocytes (PMN) are activated, and excited oxygen radicals and carbonyl groups are generated. Upon relaxation to the ground state, these molecules emit photons which can be measured as chemiluminescence (CL) using a liquid scintillation counter. Divalent cations (Mg2+, Ca2+) regenerated chemiluminescence (CL) of zymosan-stimulated human PMN abolished by 0.1 mM EGTA [ethylene glycol bis(.beta.-aminoethyl ether) N,N,N''N''-tetraacetic acid]. Phenol red, erythrocytes and free hemoglobin reduced CL. To obtain maximal CL response, the reaction mixture should contain small amounts of protein and glucose. High protein concentrations reduced CL. The test had a pH optimum of 7.1. With respect to stability, the most favorable pH was 7.4. The CL response was highly temperature-dependent, and expressed as the initial rate constant of the CL time curve it increased linearly with increasing temperatures from 20.degree. to 37.degree. C. Several factors influence the CL response by human PMN and test procedures should be standardized.