Cellular Sources of Interleukin 16 in Benign and Malignant Pleural Effusions

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.

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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

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