Study on natural killer cell activity and its characteristics during hepatocarcinogenesis in rats

Jeong, Ja-young; Lee, Kuk-kyung; Kil, Jwang-sup; Lee, Yong-soon

Korean Journal of Veterinary Research 1999;39(1):169-176.

Ja-young Jeong¹, Kuk-kyung Lee¹, Jwang-sup Kil¹, Yong-soon Lee²

¹Department of Pathology, National Institute of Toxicological Research, KFDA
²College of Veterinary Medicine, Seoul National University

랫드의 간암 발생과정에서 분리한 자연살해세포의 활성측정 및 특성연구

정자영¹, 이국경¹, 길광섭¹, 이영순²

¹식품의약품안전청 독성연구소 병리부
²서울대학교 수의과대학

Abstract

The purposes of this study were to set up the method of the natural killer(NK) cell activity assay using the flow cytometer and to examine the characteristics and distribution of the NK cell during rat hepatocarcinogenesis. Forty five male 6 week-old specific pathogen free(SPF) Sprague-Dawley rats were randomly divided into three groups. Group I was the non-treated control and given normal diet and water. Group II was treated with diethylnitrosamine(DEN, 200mg/kg, i.p.) and partial hepatectomy. Group III was treated with DEN, partial hepatectomy and 0.05% phenobarbital sodium in water from 3 to 16 weeks. All animals were examined the morphology of the large granular lymphocyte(LGL), the LGL percent of the total lymphocytes and the LGL conjugation rate with YAC-1 cell in peripheral blood, spleen and liver. Moreover, activity of the LGL isolated from peripheral blood lymphocytes was determined using the flow cytometer. As results, LGL were observed in the peripheral blood, spleen and liver. LGL were observed the relatively faintly staining basophilic cytoplasm with granules, and eccentric, often kidney-shaped nuclei in Giemsa stain. Its size was $11{sim}13{mu}m$. LGL percentage of the isolated lymphocytes in peripheral blood, spleen and liver were 1.8~2.3%, 1.3~1.4% and 0.87~0.99%, respectively. LGL conjugation rate with YAC-1 cell was shown to be peripheral blood(9.3~10.3 %) > spleen(7.7~8.7%) > liver(5.6~7.0%). The activity of the LGL isolated from peripheral blood lymphocytes in Group I, II and III was 33.7%, 30.5% and 35.4%, respectively. However, all values were not significantly between groups.

Key Words: large granular lymphocytes, natural killer cell, flow cytometer, hepatocarcinogenesis, effector:target cell conjugation assay