This article was downloaded by: [University of Aberdeen] On: 15 February 2015, At: 19:44 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK
Journal of Immunoassay and Immunochemistry Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/ljii20
SKEWNESS OF TCR Vβ OF PERIPHERAL BLOOD AND SYNOVIAL FLUID OF PATIENTS WITH RHEUMATOID ARTHRITIS a
b
a
a
Jianwei Zhou , Cui Kong , Jiang Yu , Haixin Dong , Chengqiang a
Jin & Qin Song
c
a
Clinic Laboratory, Affiliated Hospital of Jining Medical College , Jining , Shandong Province , China b
Department of Cardiovascular Disease , Affiliated Hospital of Jining Medical College , Jining , Shandong Province , China
Click for updates
c
Department of Immune Rheumatic Disease , Affiliated Hospital of Jining Medical College , Jining , Shandong Province , China Accepted author version posted online: 20 Sep 2013.Published online: 02 Dec 2013.
To cite this article: Jianwei Zhou , Cui Kong , Jiang Yu , Haixin Dong , Chengqiang Jin & Qin Song (2014) SKEWNESS OF TCR Vβ OF PERIPHERAL BLOOD AND SYNOVIAL FLUID OF PATIENTS WITH RHEUMATOID ARTHRITIS, Journal of Immunoassay and Immunochemistry, 35:2, 207-219, DOI: 10.1080/15321819.2013.841192 To link to this article: http://dx.doi.org/10.1080/15321819.2013.841192
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [University of Aberdeen] at 19:44 15 February 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Journal of Immunoassay and Immunochemistry, 35:207–219, 2014 Copyright # Taylor & Francis Group, LLC ISSN: 1532-1819 print/1532-4230 online DOI: 10.1080/15321819.2013.841192
Downloaded by [University of Aberdeen] at 19:44 15 February 2015
SKEWNESS OF TCR Vb OF PERIPHERAL BLOOD AND SYNOVIAL FLUID OF PATIENTS WITH RHEUMATOID ARTHRITIS
Jianwei Zhou,1 Cui Kong,2 Jiang Yu,1 Haixin Dong,1 Chengqiang Jin,1 and Qin Song3 1 Clinic Laboratory, Affiliated Hospital of Jining Medical College, Jining, Shandong Province, China 2 Department of Cardiovascular Disease, Affiliated Hospital of Jining Medical College, Jining, Shandong Province, China 3 Department of Immune Rheumatic Disease, Affiliated Hospital of Jining Medical College, Jining, Shandong Province, China
& To date, the complete mechanism of rheumatoid arthritis (RA) remain unclear, T cells have been proposed to play an important role in the disease initiation and progression. Presently, some researchers have reported that there were skewed TCR Vb in different samples of experimental animals or RA patients, such as in the peripheral blood, joints or synovial fluid, however, most of the results were not coincident or even conflict with each other. In this article, with real-time fluorescence quantitative PCR with DNA melting curving technique, we detected the bias of TCR Vb of RA patients, and found that although most of TCR Vb usage were different between peripheral blood and synovial fluid, the overview of all the Vb skewness was similar between the two samples. Keywords peripheral blood, rheumatoid arthritis, skewness, synovial fluid, TCR
INTRODUCTION Rheumatoid arthritis (RA) is a chronic inflammatory diseases characterized by destructive polyarthritis. Although the complete mechanism of RA remain unclear, T cells have been proposed to play an important role in the disease initiation and progression.[1,2] Mature T cells express one of two types of T cell receptors (TCRs): a heterodimer of a and b chains or c and d chains. In animal models with collagen-induced arthritis (CIA), Corthay[3] found that ab T cells play a more important role in the occurrence of RA than that of cd T cells. In the studies on rats with Address correspondence to Jianwei Zhou, Clinic Laboratory, the Affiliated Hospital of Jining Medical College, Shandong, China. E-mail: [email protected]
Downloaded by [University of Aberdeen] at 19:44 15 February 2015
208
J. Zhou et al.
CIA, researchers[4,5] found that there were TCR variable beta (Vb) genes over-expressed; moreover, accompanied by the development of the disease, some Vb families exhibited certain advantage clonetypes. In the studies on human patients with RA, T lymphocytes were considered undergoing expansion in the synovium, especially, some of TCR gene showed skewness in the joints and blood of RA patients.[6,7] If the biased TCR Vb was associated with the pathogenic T-cell populations, the corresponding TCR elements could be targeted by TCR-specific immunotherapies, such as TCR peptide vaccination or T-cell vaccination.[8,9] Unfortunately, many contradictory findings have been reported, moreover, although there are several reports relate to the bias of TCR Vb in RA patients, seldom reports focused on comparative analysis of TCR Vb gene usage of peripheral blood (PB) and synovial fluid (SF) of RA patients. More presently, real-time florescence quantitative polymerase chain reaction (FQ-PCR) and DNA melting curve analysis technique was successfully modified and used to detect the skewness of TCR.[10] In this study, we used real-time RQ-PCR and melting curve analysis technique to assay the TCR Vb gene usage of PB and SF of acute and chronic RA patients, to further analyze the clone features of these two samples, and hope to provide information for the study of mechanism. OBJECTS AND MEHTODS Screening for Controls and RA Patients Twelve healthy volunteers for providing PB samples were recruited form Health Examination Center, three patients who were operated on knees and provide SF samples came form surgery wards. All RA patients (detail information shown in Table 1) were screened according to the diagnostic criteria of the American Rheumatism Association.[11] All of objects had not been treated with immunomodulating drugs in the previous six months prior to the study and were seronegative for markers of hepatitis viruses, HIV and other pathogenic infections. Excluded from the study were
TABLE 1
The Basic Clinical Information of all the Patients with Rheumatoid Arthritis
Item
P1
P2
P3
P4
P5
P6
P7
P8
P9
P10
P11
P12
Age Sex Duration (yr) Diagnosis
39 M
Journal of Immunoassay and Immunochemistry Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/ljii20
SKEWNESS OF TCR Vβ OF PERIPHERAL BLOOD AND SYNOVIAL FLUID OF PATIENTS WITH RHEUMATOID ARTHRITIS a
b
a
a
Jianwei Zhou , Cui Kong , Jiang Yu , Haixin Dong , Chengqiang a
Jin & Qin Song
c
a
Clinic Laboratory, Affiliated Hospital of Jining Medical College , Jining , Shandong Province , China b
Department of Cardiovascular Disease , Affiliated Hospital of Jining Medical College , Jining , Shandong Province , China
Click for updates
c
Department of Immune Rheumatic Disease , Affiliated Hospital of Jining Medical College , Jining , Shandong Province , China Accepted author version posted online: 20 Sep 2013.Published online: 02 Dec 2013.
To cite this article: Jianwei Zhou , Cui Kong , Jiang Yu , Haixin Dong , Chengqiang Jin & Qin Song (2014) SKEWNESS OF TCR Vβ OF PERIPHERAL BLOOD AND SYNOVIAL FLUID OF PATIENTS WITH RHEUMATOID ARTHRITIS, Journal of Immunoassay and Immunochemistry, 35:2, 207-219, DOI: 10.1080/15321819.2013.841192 To link to this article: http://dx.doi.org/10.1080/15321819.2013.841192
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [University of Aberdeen] at 19:44 15 February 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Journal of Immunoassay and Immunochemistry, 35:207–219, 2014 Copyright # Taylor & Francis Group, LLC ISSN: 1532-1819 print/1532-4230 online DOI: 10.1080/15321819.2013.841192
Downloaded by [University of Aberdeen] at 19:44 15 February 2015
SKEWNESS OF TCR Vb OF PERIPHERAL BLOOD AND SYNOVIAL FLUID OF PATIENTS WITH RHEUMATOID ARTHRITIS
Jianwei Zhou,1 Cui Kong,2 Jiang Yu,1 Haixin Dong,1 Chengqiang Jin,1 and Qin Song3 1 Clinic Laboratory, Affiliated Hospital of Jining Medical College, Jining, Shandong Province, China 2 Department of Cardiovascular Disease, Affiliated Hospital of Jining Medical College, Jining, Shandong Province, China 3 Department of Immune Rheumatic Disease, Affiliated Hospital of Jining Medical College, Jining, Shandong Province, China
& To date, the complete mechanism of rheumatoid arthritis (RA) remain unclear, T cells have been proposed to play an important role in the disease initiation and progression. Presently, some researchers have reported that there were skewed TCR Vb in different samples of experimental animals or RA patients, such as in the peripheral blood, joints or synovial fluid, however, most of the results were not coincident or even conflict with each other. In this article, with real-time fluorescence quantitative PCR with DNA melting curving technique, we detected the bias of TCR Vb of RA patients, and found that although most of TCR Vb usage were different between peripheral blood and synovial fluid, the overview of all the Vb skewness was similar between the two samples. Keywords peripheral blood, rheumatoid arthritis, skewness, synovial fluid, TCR
INTRODUCTION Rheumatoid arthritis (RA) is a chronic inflammatory diseases characterized by destructive polyarthritis. Although the complete mechanism of RA remain unclear, T cells have been proposed to play an important role in the disease initiation and progression.[1,2] Mature T cells express one of two types of T cell receptors (TCRs): a heterodimer of a and b chains or c and d chains. In animal models with collagen-induced arthritis (CIA), Corthay[3] found that ab T cells play a more important role in the occurrence of RA than that of cd T cells. In the studies on rats with Address correspondence to Jianwei Zhou, Clinic Laboratory, the Affiliated Hospital of Jining Medical College, Shandong, China. E-mail: [email protected]
Downloaded by [University of Aberdeen] at 19:44 15 February 2015
208
J. Zhou et al.
CIA, researchers[4,5] found that there were TCR variable beta (Vb) genes over-expressed; moreover, accompanied by the development of the disease, some Vb families exhibited certain advantage clonetypes. In the studies on human patients with RA, T lymphocytes were considered undergoing expansion in the synovium, especially, some of TCR gene showed skewness in the joints and blood of RA patients.[6,7] If the biased TCR Vb was associated with the pathogenic T-cell populations, the corresponding TCR elements could be targeted by TCR-specific immunotherapies, such as TCR peptide vaccination or T-cell vaccination.[8,9] Unfortunately, many contradictory findings have been reported, moreover, although there are several reports relate to the bias of TCR Vb in RA patients, seldom reports focused on comparative analysis of TCR Vb gene usage of peripheral blood (PB) and synovial fluid (SF) of RA patients. More presently, real-time florescence quantitative polymerase chain reaction (FQ-PCR) and DNA melting curve analysis technique was successfully modified and used to detect the skewness of TCR.[10] In this study, we used real-time RQ-PCR and melting curve analysis technique to assay the TCR Vb gene usage of PB and SF of acute and chronic RA patients, to further analyze the clone features of these two samples, and hope to provide information for the study of mechanism. OBJECTS AND MEHTODS Screening for Controls and RA Patients Twelve healthy volunteers for providing PB samples were recruited form Health Examination Center, three patients who were operated on knees and provide SF samples came form surgery wards. All RA patients (detail information shown in Table 1) were screened according to the diagnostic criteria of the American Rheumatism Association.[11] All of objects had not been treated with immunomodulating drugs in the previous six months prior to the study and were seronegative for markers of hepatitis viruses, HIV and other pathogenic infections. Excluded from the study were
TABLE 1
The Basic Clinical Information of all the Patients with Rheumatoid Arthritis
Item
P1
P2
P3
P4
P5
P6
P7
P8
P9
P10
P11
P12
Age Sex Duration (yr) Diagnosis
39 M
