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Stereological and Flow Cytometry Characterization of Leukocyte Subpopulations in Models of Transient or Permanent Cerebral Ischemia
Published on: December 28, 2014
Polymorphonuclear leukocyte behavior in a nonhuman primate focal ischemia model
J A Ember1, G J del Zoppo, E Mori
1Department of Immunology, Scripps Research Institute, La Jolla, CA 92037.
Abstract:
There is increasing interest in the role of polymorphonuclear (PMN) leukocytes in the evolution of focal cerebral infarction. Surgical preparation of focal cerebral ischemia models may alter leukocyte reactivity and thereby make interpretation of leukocyte function following ischemia/reperfusion difficult. The effects of surgical preparation and of experimental ischemia/reperfusion on granulocyte function have been examined prospectively in a baboon model. Twenty-six adolescent male baboons underwent surgical preparation, of which 21 underwent middle cerebral artery occlusion/reperfusion. Four additional animals served as nonsurgical controls. Peripheral venous blood specimens were taken for performing assays of leukocyte function at defined intervals before and after both the surgical preparation (i.e., the overall procedure for implantation of the middle cerebral artery occlusion device) and occlusion/reperfusion. A stress-related elevation in total leukocyte number was attributed mainly to an increase in the number of circulating PMN leukocytes. Values rose from 13.9 +/- 4.9 x 10(3) to 27.8 +/- 5.8 x 10(3)/microliters, (+/- SD; n = 21) for total leukocyte number, with p < 0.001, and from 4.3 +/- 2.1 x 10(3) to 15.9 +/- 4.7 x 10(3)/microliters (n = 21) for PMN leukocytes, with p < 0.001. Surgical preparation had no effect (p > or = 0.4) on the ability of PMN leukocytes, isolated 24 h after the implantation procedure, to display polarization, O2.- production, or beta-glucuronidase release when stimulated with human C5a. A moderate decrease in the chemotactic response to C5a resolved within the 7-day postsurgery (preocclusion) period. Three-hour middle cerebral artery occlusion and 1-h reperfusion resulted in a significant reduction in C5a-induced polarization.(ABSTRACT TRUNCATED AT 250 WORDS)
Insights
Surgical preparation did not affect polymorphonuclear (PMN) leukocyte function in baboons. However, middle cerebral artery occlusion/reperfusion significantly reduced PMN leukocyte polarization, impacting stroke research.
Area of Science:
- Neuroscience
- Immunology
- Vascular Biology
Background:
- Polymorphonuclear (PMN) leukocytes play a role in focal cerebral infarction.
- Surgical preparation for ischemia models may affect leukocyte reactivity, complicating research interpretation.
Purpose of the Study:
- To prospectively examine the effects of surgical preparation and ischemia/reperfusion on granulocyte function in a baboon model.
- To determine if surgical procedures alter PMN leukocyte responses relevant to stroke research.
Main Methods:
- Baboons underwent surgical preparation for middle cerebral artery occlusion/reperfusion.
- Leukocyte function assays (polarization, O2.- production, beta-glucuronidase release, chemotaxis) were performed pre- and post-procedure.
- Comparison between nonsurgical controls, surgically prepared animals, and those undergoing occlusion/reperfusion.
Main Results:
- Surgical preparation caused a stress-related increase in circulating PMN leukocytes (p < 0.001).
- Surgical preparation alone did not impair PMN leukocyte function (polarization, O2.- production, beta-glucuronidase release) 24h post-op (p >= 0.4).
- Middle cerebral artery occlusion/reperfusion significantly reduced C5a-induced PMN leukocyte polarization.
Conclusions:
- Surgical preparation in baboons does not significantly alter key PMN leukocyte functions relevant to ischemia/reperfusion studies.
- Ischemia/reperfusion itself, however, does impact PMN leukocyte function, specifically polarization.
- Findings suggest careful consideration of the ischemia/reperfusion model's direct effects on leukocytes is necessary for accurate stroke research.

