Influences of Heat on Function and Morphology of Peripheral Blood Cells

Choi, Jong Weon; Nahm, Chung Hyun; Lee, Jong Woog; Kim, Jin Ju; Kim, Soon Ki; Pai, Soo Hwan

Abstract

BACKGROUND: Heat-treated erythrocytes have been reported to be undergone fragmentation and microspherocytes transformation in vitro. However, the changes for enzyme activity of WBCs is not well known. The purpose of this study is to investigate the effects of temperature on function and morphology of WBCs and platelets. And we also defined the relationship between temperature-induced changes in morphology of the erythrocytes and cholesterol level.
METHODS
Peripheral blood specimens collected from 55 normal healthy subjects were assayed. CBC and biochemical test for heat-treated specimens were performed. Osmotic fragility was tested and echinocytes were counted using phase contrast microscope. DNA was extracted from heated WBCs with Wizard genomic DNA extraction kit(Promega Corp, USA) and applied on 1.5% agarose gel. After heating, plasma proteins were fractionated by sodium dodecyl sulfate-poly-acrylamide gel electrophoresis (SDS-PAGE) and compared according to changes of temperature. For the phagocytic function of WBCs, 1 x10(7) CFU/mL of gram-negative bacilli were incubated with specimens for 4 hours. Enzymatic activities for myeloperoxidase(MPO), alkaline phosphatase (ALP), and esterase (EST) were confirmed with cytochemical reaction.
RESULTS
After incubation at 50 degrees C for 5 min, platelets were counted as 2,978.2 x 10(9)/L and the number of eosinophil was 5.58 x 10(9)/L. And blood glucose level was decreased from 92.4 mg/dL to 59.1 mg/dL, whereas the value of cholesterol was changed as follows; 201.3 mg/dL at 0 sec, 174.6 mg/dL at 60 sec, 151.2 mg/dL at 90 sec, and 197.6 mg/dL at 5 min. In the same temperature, the proportion of echinocytes was 94.1% at 90 sec and decreased to 2.3% at 5 min. In osmotic fragility test, the hemolysis of RBCs began at 0.74% of NaCl and completed at 0.35% of NaCl. Phagocytic function of leukocytes was lost at 52 degrees C, and enzyme activity was lost at following temperature ; MPO at 70t, EST at 70 degrees C, and ALP at 56 degrees C, respectively. SDS-PAGE patterns reveal individual differences of protein at same temperature condition. Aggregating function of platelets were lost, after incubation at 43 degrees C for 5 min. In EDTA-anticoagulated blood, temperature-induced platelelet aggregation was intensified, while in heparinized blood, platelet aggregation was not induced by heat.
CONCLUSION
For morphological transformation of erythrocytes in vitro, 50 degrees C is critical point of temperature to induce fragmentation and microspherocytes. Plasma cholesterol appears to play some role to formation of echinocytes induced by heat. MPO is more stable for high temperature than EST and ALP. And sensitivity of RBCs to temperature is considered to be different individually.

Influences of Heat on Function and Morphology of Peripheral Blood Cells

Abstract

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