Presence of Oligoclonal T Cells in Cerebrospinal Fluid of a Child with Multiphasic Disseminated Encephalomyelitis following Hepatitis A Virus Infection

Oleszak, Emilia L.; Lin, Wan Lu; Legido, Agustín; Melvin, Joseph J.; Hardison, H. Huntley; Hoffman, Brad E.; Katsetos, Christos D.; Platsoucas, Chris D.

doi:10.1128/cdli.8.5.984-992.2001

Cited by 33 publications

(46 citation statements)

References 71 publications

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“…cDNA was synthesized from total RNA and primed with oligo-(dT) 15 -NotI (Promega), using SuperScript II (Gibco/Brl), following the manufacturer's instructions (32)(33)(34)(35)(36)(37). Double-stranded cDNA was blunt ended for efficient adaptor ligation by adding T4 DNA polymerase.…”

Section: Synthesis Of Cdnamentioning

confidence: 99%

“…The blunt-ended cDNA was ligated at both the 59 and 39 blunt ends with an equivalent molar concentration of a nonpalindromic adaptor (NPA) (32)(33)(34)(35)(36)(37) by incubation for 14 h at 16˚C with T4 DNA ligase (Life Technologies-BRL). This adaptor, a modification of the one previously described (32)(33)(34)(35)(36)(37), consists of two oligonucleotides (Supplemental Table I), the 59-AATTCGAACCCCTTCGAGAA-TGCG-39 and its complementary 39-GCTTGGGGAAGCTCTTACGC-p-59, preannealed to each other. The ligated adaptor was removed from the 39 end of the double-stranded cDNA by digestion with NotI restriction endonuclease, as described (32)(33)(34)(35)(36)(37), while it remained in the 59 end.…”

Section: Synthesis Of Cdnamentioning

confidence: 99%

“…This adaptor, a modification of the one previously described (32)(33)(34)(35)(36)(37), consists of two oligonucleotides (Supplemental Table I), the 59-AATTCGAACCCCTTCGAGAA-TGCG-39 and its complementary 39-GCTTGGGGAAGCTCTTACGC-p-59, preannealed to each other. The ligated adaptor was removed from the 39 end of the double-stranded cDNA by digestion with NotI restriction endonuclease, as described (32)(33)(34)(35)(36)(37), while it remained in the 59 end. The digested cDNA was purified using a G-50 column (59-39, Boulder, CO), as recommended by the manufacturer.…”

Section: Synthesis Of Cdnamentioning

confidence: 99%

“…First cycle of amplification by NPA-PCR. NPA-PCR was carried out as previously described (32)(33)(34)(35)(36)(37), with minor modifications. The NPA 59-AATTCGAACCCCTTCGAGAATGCT-39 was used as the 59 amplification primer.…”

Section: Synthesis Of Cdnamentioning

confidence: 99%

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Aneurysmal Lesions of Patients with Abdominal Aortic Aneurysm Contain Clonally Expanded T Cells

White

Lin

et al. 2014

The Journal of Immunology

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Abdominal aortic aneurysm (AAA) is a common disease with often life-threatening consequences. This vascular disorder is responsible for 1–2% of all deaths in men aged 65 years or older. Autoimmunity may be responsible for the pathogenesis of AAA. Although it is well documented that infiltrating T cells are essentially always present in AAA lesions, little is known about their role in the initiation and/or progression of the disease. To determine whether T cells infiltrating AAA lesions contain clonally expanded populations of T cells, we amplified β-chain TCR transcripts by the nonpalindromic adaptor–PCR/Vβ-specific PCR and/or Vβ-specific PCR, followed by cloning and sequencing. We report in this article that aortic abdominal aneurysmal lesions from 8 of 10 patients with AAA contained oligoclonal populations of T cells. Multiple identical copies of β-chain TCR transcripts were identified in these patients. These clonal expansions are statistically significant. These results demonstrate that αβ TCR+ T lymphocytes infiltrating aneurysmal lesions of patients with AAA have undergone proliferation and clonal expansion in vivo at the site of the aneurysmal lesion, in response to unidentified self- or nonself Ags. This evidence supports the hypothesis that AAA is a specific Ag–driven T cell disease.

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Section: Synthesis Of Cdnamentioning

confidence: 99%

Section: Synthesis Of Cdnamentioning

confidence: 99%

Section: Synthesis Of Cdnamentioning

confidence: 99%

Section: Synthesis Of Cdnamentioning

confidence: 99%

See 2 more Smart Citations

Aneurysmal Lesions of Patients with Abdominal Aortic Aneurysm Contain Clonally Expanded T Cells

White

Lin

et al. 2014

The Journal of Immunology

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“…Among the advantages of this methodology is the fact that only one 5' end extension primer is required. Since it was first described, this methodology has been successfully used to clone the genes encoding TCR transcripts of T cells isolated from humans and mice (8)(9)(10)(11)(12).…”

Section: Introductionmentioning

confidence: 99%

The non-palindromic adaptor-PCR method for the identification of the T-cell receptor genes of an interferon-gamma-secreting T-cell hybridomaspecific for trans-sialidase, an immunodominant Trypanosoma cruzi antigen

Hiyane

Boscardin

Rodrigues

2006

Braz J Med Biol Res

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Cloning of the T-cell receptor genes is a critical step when generating T-cell receptor transgenic mice. Because T-cell receptor molecules are clonotypical, isolation of their genes requires reverse transcriptaseassisted PCR using primers specific for each different Vα or Vß genes or by the screening of cDNA libraries generated from RNA obtained from each individual T-cell clone. Although feasible, these approaches are laborious and costly. The aim of the present study was to test the application of the non-palindromic adaptor-PCR method as an alternative to isolate the genes encoding the T-cell receptor of an antigenspecific T-cell hybridoma. For this purpose, we established hybridomas specific for trans-sialidase, an immunodominant Trypanosoma cruzi antigen. These T-cell hybridomas were characterized with regard to their ability to secrete interferon-γ, IL-4, and IL-10 after stimulation with the antigen. A CD3 + , CD4 + , CD8 -interferon-γ-producing hybridoma was selected for the identification of the variable regions of the T-cell receptor by the non-palindromic adaptor-PCR method. Using this methodology, we were able to rapidly and efficiently determine the variable regions of both T-cell receptor chains. The results obtained by the non-palindromic adaptor-PCR method were confirmed by the isolation and sequencing of the complete cDNA genes and by the recognition with a specific antibody against the T-cell receptor variable ß chain. We conclude that the nonpalindromic adaptor-PCR method can be a valuable tool for the identification of the T-cell receptor transcripts of T-cell hybridomas and may facilitate the generation of T-cell receptor transgenic mice.

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Acute Disseminated Encephalomyelitis and Related Conditions

Kálmán¹,

Rust

2007

Neuroimmunology in Clinical Practice

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Presence of Oligoclonal T Cells in Cerebrospinal Fluid of a Child with Multiphasic Disseminated Encephalomyelitis following Hepatitis A Virus Infection

Cited by 33 publications

References 71 publications

Aneurysmal Lesions of Patients with Abdominal Aortic Aneurysm Contain Clonally Expanded T Cells

Aneurysmal Lesions of Patients with Abdominal Aortic Aneurysm Contain Clonally Expanded T Cells

The non-palindromic adaptor-PCR method for the identification of the T-cell receptor genes of an interferon-gamma-secreting T-cell hybridomaspecific for trans-sialidase, an immunodominant Trypanosoma cruzi antigen

Acute Disseminated Encephalomyelitis and Related Conditions

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