Cellular Sources of Interleukin 16 in Benign and Malignant Pleural Effusions
Total Page:16
File Type:pdf, Size:1020Kb
Cellular sources of interleukin 16 in benign and malignant pleural effusions LI Jian-jun W E I W e i SHI Huan-zhong LI Ying-xin MO Wu-ning interleukin 16;.flow cytometry; immunohistochemistry; pleural disease Background Interleukin 16 (IL-16) can be detected by ELISA in pleural effusion (PE) and its concentration is higher than in serum. This study investigated the cellular sources of IL-16 in PE. Methods The samples of PE were collected from 34 patients who were newly diagnosed having PE in the pleural cavity. We performed cell culture to purify the pleural mesothelial cells (PMC), Wright staining to count the purity and immu nocytochemic al stain to i dentify the cultured cell s. The intracellular IL-16 expression was detected by flow cytometry (FCM). The different cells in PE were first separated by magnetic cell sorting (MCAS) then the separated cells were cultured in RPMI1640 with 10% fetal calf serum (FCS). We extracted the supernatant and detected IL-16 concentration by ELISA. The IL-16 protein was detected by immunohistochemistry and double immunofluorescence staining. Results The percentages of cells which secreted IL-16 were: CD3+CD8 cells ((74.27±15.56)%, n=34); CD3+CD8+ cells ((69.86±18.55)%, n=34); CD19+ cells ((45.30±18.77)%, n=15); CD14+ cells ((16.91±16.69)%, n=15); and PMC ((2. 05±1.85)%, n=7). The co ncentrations of IL-16 in the supernatant from cultured cells were: CD4+ cells ((102.50±42.51) ng/L , n=5); CD8+ cells ((92.5 8±18.34) ng /L, n=5); CD1 9+ cells ((79.85±5.62) ng/L, n=5); CD14+ cells ((58.51±25.38) ng/L, n=5); and PMC ((18.14±8.37) ng/L, n=5). In lymphocytes, monocytes/macrophages and PMC, we could observe the cells that expressed IL-16 protein. In paraffin-embedded sections, we also could observe by immunohistochemistry the CD4+IL-16+ cells, CD8+IL-16+ cells, CD19+IL-16+ cells, and CD14+IL-16+ cells. Conclusions IL-16 in PE is mainly secreted by T lymphocytes, including CD3+CD8- cells and CD3+CD8+celIs. CD19+ cells and CD14+ cells can also secrete IL-16, but the percentage of PMC that can secrete IL-16 is very low. 10.3 760/cma.j.issn.0366-6999.2011.24.008 Ma ster Degree C andidate, Fi rst Affiliate d Hospital, Guangxi Me dical Univers ity, Nanning, Guangxi 530021, China (Li JJ and Wei W) Department of Respiratory Diseases, Union Hospital, Tongji Me dical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, China (Shi HZ) La boratory Cent er, Pharmace utical Colleg e of Guangxi Medical University, Nanning, Guangxi 530021, China (Li YX) La boratory Cent er, First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi 530021, China (Mo WN) <作者简 介>=Correspondence to: Dr. MO Wu-ning, Laboratory Center, First Af filiated Hospital, Guangxi Medical University, Nanning, Guangxi 530021, China (Tel: 86-771-5356052. Fax: 86-771-5356052. Email: mown 163 @ 163.com) LI Jian-jun is now working in Department of AIDS/STD, Guangxi Ce nter for Dise ase Control and Preventio n, Nanning, Guangxi 530028, China. WE I Wei is now working in First Affiliated Hospital to Guangxi College of Traditional Chinese Medicine, Nanning, Guangxi 530028, China. This stud y was suppor ted by grants from the Gu angxi Medical University, Dr. Start Fund (No. 304201) and the Guangxi Health Department issues (No. Z2006062). None of the authors have a financial relationship with a commercia l entity tha t has an interest in the subject of this manuscript. 4 1 6 1 4162 4163 @ @1. Cen ter DM, Cr uik sha nk W. Mo dula tio n of lymphocyte migration by human lymphokines. 1. Identification and characterization of chemoattractant activity for lymphocytes from mitogen stimulated mononuclear cells. J Immunol 1982; 128: 2563-2568. @@2. Arima M, Plitt J, Stcllato C, Bickel C, Motojima S, Makino S, et al. Expression of interleukin-16 by human epithelial cells inhibition by dexamethasone. Am J Respir Cell Mol Biol 1999; 21: 684-692. @@3. Krautwald S. IL-16 activates the SAPK signaling pathway in 3806-3814. CD4+ macrophage. J Immnol 1998; 160: 5874-5879. @@10. Elssner A, Doseff AI, Duncan M, Kotur M, Wewers MD. @ @4. Mathy N L, Bannert N , Norley SG, Kurth R. Cutting edge: IL-16 is constitutively present in peripheral blood monocytes CD4 is not required for the functional activity of IL-16. J and spontane-ousl y released d uring apoptosis. J Immunol Immnol 2000; 164: 4429-4432. 2004; 172: 7721-7725. @@5. Laberge S, Pinsonneault S, Ernst P, Olivenstein R, Ghaffar O, @@11. Qin XJ, Shi HZ, Huang ZX, Kang LF, Mo WN, Wu C. Center D M, et al. Ph enotype of IL-16 producing cells in Interleukin-16 in tuberculous and malignant pleural effusions. bronchia l mucosa: ev idence for the human eosinophil and Eur Respir J 2005; 25:605-611. mast cell as cellular sources of IL-16 in asthma. Int Arch @@12. Pal a P, Hussel l T, Openshaw PJ. Flow cytometrick Allergy Immnology 1999; 119: 120-125. meas urement of intracellular cytokines. J Immunol Methods @ @6. Krug N, Cruikshank WW, Tschernig T, Erpenbeck VJ, Balke 2000; 243: 107-124. K, Hohl feld JM, et al. Interleukin 16 and T-cell @@13. Sewell WA, North ME, Webster DB, Farrant J. Determination chemoattractant activity in bronchial-veolarlavage 24 hours of i ntracellular cytokine by flow-cytometry following after allergen challenge in asthma. Am J Respir Crit Care whole-blood culture. J Immunol Methods 1997; 209: 67-74. Med 2000; 162: 105-111. @@14. Katayama H, Yokoyama A, Kohno N, Sakai K, Hiwada K, @@7. Kaser A, Dunzendorfer S, Offner FA, Ludwiczek O, Enrich B, Yama da H, et al. Production of eosinophilic chemokines by Koch RO, et al. B lymphocyte-derived IL-16 attracts normal pleural mesothelial cells. Am J Respir Cell Mol Biol dendritic cells and Th cells. J Immunol 2000; 165: 2002; 26: 398-404. 2474-2480. @@15. Fra ncé s R, Rodr iguez E, Mun oz C, Zapater P, De la ML, @@8. Reich K, Heine A, Hugo S, Blaschke V, Middel P, Kaser A, et Ndongo M, et al. Intracellular cytokine expression in ai. Engagement of the Fc epsilon RI stimulates the production peritoneal monocyte/macrophages obtained from patients of IL-16 in langerhans cell-like dendritic cells. J Immunol with cirrhosis and presence of bacterial DNA. Eur J 2001; 167: 6321-6329. Gastroenteroi Hepatol 2005; 17:45-51. @ @9. Sciaky D, Brazer W, Center DM, Cruikshank WW, Smith TJ. Cultured human fibroblasts express constitutive IL-16mRNA: cytokine induction of active IL-16 protein synthesis through a 2011-05-08 caspase-3-dependent mechanism. J Immunol 2000; 164: Cellular sources of interleukin 16 in benign and malignant pleural effusions 作者: LI Jian-jun, WEI Wei, SHI Huan-zhong, LI Ying-xin, MO Wu-ning 作者单位: LI Jian-jun(Department of AIDS/STD, Guangxi Center for Disease Control and Prevention, Nanning, Guangxi 530028, China), WEI Wei(First Affiliated Hospital to Guangxi College of Traditional Chinese Medicine, Nanning, Guangxi530028, China), SHI Huan-zhong(Department of Respiratory Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology,Wuhan, Hubei 430022, China), LI Ying-xin(Laboratory Center, Pharmaceutical College of Guangxi Medical University, Nanning, Guangxi 530021, China(), MO Wu-ning(Laboratory Center, First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi 530021, China) 刊名: 中华医学杂志(英文版) 英文刊名: Chinese Medical Journal 年,卷(期): 2011,124(24) 参考文献(15条) 1.Sciaky D;Brazer W;Center DM;Cruikshank WW Smith TJ Cultured human fibroblasts express constitutive IL-16mRNA:cytokine induction of active IL-16 protein synthesis through a caspase-3-dependent mechanism 2000 2.Qin XJ;Shi HZ;Huang ZX;Kang LF Mo WN Wu C Interleukin-16 in tuberculous and malignant pleural effusions 2005 3.Elssner A;Doseff AI;Duncan M;Kotur M Wewers MD IL-16 is constitutively present in peripheral blood monocytes and spontane- ously released during apoptosis 2004 4.Center DM;Cruikshank W Modulation of lymphocyte migration by human lymphokines,1.Identification and characterization of chemoattractant activity for lymphocytesfrom mitogen stimulated mononuclear cells 1982 5.Reich K;Heine A;Hugo S;Blaschke V Middel P Kaser A et ai Engagement of the Fc epsilon RI stimulates the production of IL-16 in langerhans cell-like dendritic cells 2001 6.Kaser A;Dunzendorfer S;Offner FA;Ludwiczek O Enrich B Koch RO B lymphocyte-derived IL-16 attracts dendritic cells and Th cells 2000 7.Krug N;Cruikshank WW;Tschernig T;Erpenbeck VJ Balke K Hohlfeld JM Interleukin 16 and T-cell chemoattractant activity in bronchial-veolarlavage 24 hours after allergen challenge in asthma 2000 8.Laberge S;Pinsonneault S;Ernst P;Olivenstein R Ghaffar O Center DM Phenotype of IL-16 producing cells in bronchial mucosa:evidence for the human eosinophil and mast cell as cellular sources of IL-16 in asthma 1999 9.Mathy NL;Bannert N;Norley SG;Kurth R Cutting edge:CD4 is not required for the functional activity of IL-16 2000 10.Krautwald S IL-16 activates the SAPK signaling pathway in CD4+ macrophage 1998 11.Arima M;Plitt J;Stcllato C;Bickel C Motojima S Makino S Expression of interleukin-16 by human epithelial cells inhibition by dexamethasone 1999 12.Francés R;Rodriguez E;Munoz C;Zapater P De la ML Ndongo M Intracellular cytokine expression in peritoneal monocyte/macrophages obtained from patients with cirrhosis and presence of bacterial DNA 2005 13.Katayama H;Yokoyama A;Kohno N;Sakai K Hiwada K Yamada H Production of eosinophilic chemokines by normal pleural mesothelial cells 2002 14.Sewell WA;North ME;Webster DB;Farrant J Determination of intracellular cytokine by flow-cytometry following whole-blood culture 1997 15.Pala P;Hussell T;Openshaw PJ Flow cytometrick measurement of intracellular cytokines 2000 本文读者也读过(10条) 1.