Detection of cell specific cluster determinant expression by reverse transcriptase polymerase chain reaction

James-Yarish, Michelle; Bradley, W G; Emmanuel, Patricia; Good, Robert A.; Day, Noorbibi K.

doi:10.1016/0022-1759(94)90126-0

Cited by 16 publications

(10 citation statements)

References 10 publications

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“…Expression of the CD genes in Pal‐1 cells was also examined by RT‐PCR essentially as previously described (James‐Yarish et al , 1994). The primer sequences specific for CD2, CD3‐γ, CD7, CD19 and CD20 genes were obtained from GenBank and a previous report (James‐Yarish et al , 1994). The experiments were controlled by the use of two cell lines, BALL‐1 (B‐cell leukaemia) (Hiraki et al , 1977) and TALL‐1 (T‐cell leukaemia) (Hiraki et al , 1978).…”

Section: Resultsmentioning

confidence: 99%

Epstein–Barr virus (EBV)‐positive pyothorax‐associated lymphoma (PAL): chromosomal integration of EBV in a novel CD2‐positive PAL B‐cell line

Daibata¹,

Taguchi²,

Nemoto³

et al. 2002

Br J Haematol

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Summary. Pyothorax-associated lymphoma (PAL) is a clinico-pathological entity arising in the pleural cavity of patients with long-standing inflammatory pyothorax. PAL is closely associated with Epstein-Barr virus (EBV), but how this virus contributes to the development of the lymphoma is unknown. We have successfully obtained a novel EBV-infected PAL cell line, designated Pal-1. The cell line and its source coexpressed CD2 and CD20 molecules, but other representative B-and T-cell markers such as CD1, CD3, CD5, CD7, CD10 and CD19 were not found. The B-cell origin of Pal-1 cells was proven by rearrangement of the immunoglobulin heavy-and light-chain genes without rearranged T-cell receptor genes. Both the cell line and primary tumour cells carried monoclonal EBV genome. Although EBV genome is known to be maintained as circular extrachromosomal DNA, neither circular nor linear extrachromosomal EBV DNA was detectable in Pal-1 cells by in situ lysis gel analysis. Fluorescence in situ hybridization demonstrated viral integration at a marker chromosome mostly consisting of the centromere region of chromosome 1. The viral integration event may enhance a chromosomal instability at the insertion site. This cell line represents the first example of EBV integration in PAL and could enable the study of the potential role of integrated viral infection in the development of PAL as well as mechanism of the aberrant phenotype expression.

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Section: Resultsmentioning

confidence: 99%

Epstein–Barr virus (EBV)‐positive pyothorax‐associated lymphoma (PAL): chromosomal integration of EBV in a novel CD2‐positive PAL B‐cell line

Daibata¹,

Taguchi²,

Nemoto³

et al. 2002

Br J Haematol

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“…13 The CD4 ϩ CD28 null phenotype was further confirmed by reversetranscriptase polymerase chain reaction (RT-PCR) analysis of mRNA for the CD4 and CD28 markers using primers for CD4 (5Ј-TGGCTCTGGAAACCTCACCCT-3Ј and 5Ј-GGCACTGGCAGG-TCTTCTTCT-3Ј) and CD28 (5Ј-GTTTGAGTGCCTTGATCATGT-GC-3Ј and 5Ј-GGCGATCTGCTTCACCAAAATC-3Ј). 14,15 Blocking MHC Class I and II Antigen Presentation MHC-restricted antigen presentation was assessed by incubating the PBMCs with blocking antibodies for 40 minutes at room temperature before antigen stimulation. The antibodies used were 10 g/mL of anti-human MHC-class I (HLA A, B, and C; Dako), anti-human CD4, anti-human MHC class II (HLA DR, DP, DQ), or a combination of both anti-CD4 and anti-human class II antibodies (Becton Dickinson).…”

Section: Reverse-transcriptase Polymerase Chain Reactionmentioning

confidence: 99%

Heat-Shock Protein 60-Reactive CD4 ⁺ CD28 ^null T Cells in Patients With Acute Coronary Syndromes

et al. 2004

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Background-CD4ϩ CD28 null T cells are present in increased numbers in the peripheral blood of patients with acute coronary syndrome (ACS) compared with patients with chronic stable angina (CSA). The triggers of activation and expansion of these cells to date remain unclear. Methods and Results-Twenty-one patients with ACS and 12 CSA patients with angiographically confirmed coronary artery disease and 9 healthy volunteers were investigated. Peripheral blood leukocytes were stimulated with human cytomegalovirus (HCMV), Chlamydia pneumoniae, human heat-shock protein 60 (hHSP60), or oxidized LDL (ox-LDL). CD4 ϩ CD28 null cells were separated by flow cytometry and assessed for antigen recognition using upregulation of interferon-␥ and perforin mRNA transcription as criteria for activation. CD4ϩ CD28 null cells from 12 of 21 patients with ACS reacted with hHSP60. No response was detected to HCMV, C pneumoniae, or ox-LDL. Incubation of the cells with anti-MHC class II and anti-CD4 antibodies but not anti-class I antibodies blocked antigen presentation, confirming recognition of the hHSP60 to be via the MHC class II pathway. Patients with CSA had low numbers of CD4 ϩ CD28 null cells. These cells were nonreactive to any of the antigens used. Circulating CD4 ϩ CD28 null cells were present in 5 of the 9 healthy controls. None reacted with hHSP60. Conclusions-We have shown that hHSP60 is an antigen recognized by CD4ϩ CD28 null T cells of ACS patients. Endothelial cells express hHSP60 either constitutively or under stress conditions. Circulating hHSP60-specific CD4 ϩ CD28 null cells may, along other inflammatory mechanisms, contribute to vascular damage in these patients.

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“…PBMCs were used as positive controls. The primers used for PCR were as follows; CD25, forward 5Ј-ACAACCAATGTCAATGCACAAGCT-3Ј and reverse 5Ј-TCTGTTCCCGGCTTCTTACCAAGA-3Ј [19]; CD122, forward 5Ј-CCGTGGCTCGGCCACCTC-3Ј and reverse 5Ј-TAGGGGTCGTAAGTA-AAGTACACC-3Ј [13]; CD132, forward 5Ј-CCAGGACCCACGGGAACCCA-3Ј and reverse 5Ј-GGTGGGAATTCGGGGCATCG-3Ј [11]; and human glyceraldehyde-3-phosphate dehydrogenase (GAPDH), forward 5Ј-TGAAGGTCG-GAGTCAACGGATTTGGT-3Ј and reverse 5Ј-CATGTGGGCCAATGAGGTC-CACCAC-3Ј [4]. The primers for CD25, CD122, CD132, and GAPDH were constructed to generate fragments of 634/419, 437, 493, and 983 bp, respectively.…”

Section: Reverse Transcriptase-polymerase Chain Reaction (Rt-pcr)mentioning

confidence: 99%

Expression of IL-2 receptor β and γ chains by human gingival fibroblasts and up-regulation of adhesion to neutrophils in response to IL-2

Ozawa

Tada

Tamai

et al. 2003

Journal of Leukocyte Biology

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To investigate the role of human gingival fibroblasts (HGF), the major constituents of gingival tissue in periodontal inflammatory disease, the expression of interleukin-2 receptor (IL-2R) alpha, beta, and gamma chains was examined. Immunohistochemistry showed a pronounced accumulation of CD8(+) T cells in the inflamed lamina propria of gingival tissue from patients with adult periodontitis. HGF express IL-2Rbeta and IL-2Rgamma at mRNA and protein levels, but the expression of IL-2Ralpha could not be detected, as assessed by reverse transcriptase-polymerase chain reaction and flow cytometry. IL-2Rbeta, and -gamma expressed on HGF were functionally active, as addition of neutralizing anti-IL-2Rbeta and -gamma antibodies caused inhibition of the IL-2-induced production of monocyte chemoattractant protein-1 (MCP-1), and addition of IL-2 induced phosphorylation of Janus tyrosine kinase 3, which is critical in signaling through IL-2Rgamma in HGF. The IL-2-induced MCP-1 production was significantly inhibited by pretreatment with neutralizing antibody to IL-15. Addition of IL-2 also induced a marked up-regulation of the expression of intercellular adhesion molecule-1 (ICAM-1) on the surface of HGF, which in turn, significantly augmented the adhesion of human neutrophils, which were inhibited by an anti-ICAM-1 antibody. These results suggest that HGF express functional IL-2Rbetagamma, respond to IL-2 from infiltrated T cells, and actively participate in the inflammatory process in the periodontal region and that IL-15 produced by HGF sustains IL-2-mediated signaling in HGF.

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Detection of cell specific cluster determinant expression by reverse transcriptase polymerase chain reaction

Cited by 16 publications

References 10 publications

Epstein–Barr virus (EBV)‐positive pyothorax‐associated lymphoma (PAL): chromosomal integration of EBV in a novel CD2‐positive PAL B‐cell line

Epstein–Barr virus (EBV)‐positive pyothorax‐associated lymphoma (PAL): chromosomal integration of EBV in a novel CD2‐positive PAL B‐cell line

Heat-Shock Protein 60-Reactive CD4 ⁺ CD28 ^null T Cells in Patients With Acute Coronary Syndromes

Expression of IL-2 receptor β and γ chains by human gingival fibroblasts and up-regulation of adhesion to neutrophils in response to IL-2

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Detection of cell specific cluster determinant expression by reverse transcriptase polymerase chain reaction

Cited by 16 publications

References 10 publications

Epstein–Barr virus (EBV)‐positive pyothorax‐associated lymphoma (PAL): chromosomal integration of EBV in a novel CD2‐positive PAL B‐cell line

Epstein–Barr virus (EBV)‐positive pyothorax‐associated lymphoma (PAL): chromosomal integration of EBV in a novel CD2‐positive PAL B‐cell line

Heat-Shock Protein 60-Reactive CD4 + CD28 null T Cells in Patients With Acute Coronary Syndromes

Expression of IL-2 receptor β and γ chains by human gingival fibroblasts and up-regulation of adhesion to neutrophils in response to IL-2

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Heat-Shock Protein 60-Reactive CD4 ⁺ CD28 ^null T Cells in Patients With Acute Coronary Syndromes