Purified protein derivative of Mycobacterium tuberculosis and excretory-secretory antigen(s) of Toxocara canis expand in vitro human T cells with stable and opposite (type 1 T helper or type 2 T helper) profile of cytokine production.

Prete, G.; Carli, Marco De; Mastromauro, C; Biagiotti, R; Macchia, Donatella; Falagiani, P.; Ricci, M; Romagnani, Sergio

doi:10.1172/jci115300

Cited by 654 publications

(370 citation statements)

References 22 publications

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“…Similar subsets have been identified in humans [3,4], but in contrast to the mouse, production of IL-2, IL-10 and IL-13 is shared by both classes of Th cells.…”

Section: Introductionsupporting

confidence: 62%

Intracellular cytokine expression in whole blood preparations from normals and patients with atopic dermatitis

Ferry

Antrobus

Hužička

et al. 1997

Clinical and Experimental Immunology

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SUMMARYIn recent years, the importance of characterizing the role of cytokines in a wide range of clinical conditions has resulted in development of new methods to assess cytokine expression in clinical samples. The use of anti-cytokine MoAbs and flow cytometry to detect cytokines intracellularly at the single-cell level has the potential to quantify cytokine production in different diseases. For this technique to be useful in a clinical setting, rapid throughput of clinical samples and a cheap, reliable assay would be required, therefore the development of the above technique using unseparated whole blood samples would be advantageous. Using this technique, only one study to date (Maino et al., 1996) has used unseparated whole blood as the source of cells for detecting intracellular cytokines. In clinical practice, whole blood may be optimal, since this most closely approximates conditions in vivo: as no purification of blood mononuclear cells is required, very little blood is needed to detect a number of cytokines simultaneously in various lymphocyte subpopulations, and the assay can be applied to samples from infants and children. In this study we describe an intracellular cytokine assay using unseparated whole blood from normals. In activated CD8 ¹ T cells, IL-2 and interferon-gamma (IFN-g) were optimally induced after 10 h stimulation with phorbol 12-myristate acetate (PMA)/ionomycin, and in CD8 þ T cells IL-2 was optimally induced after 10 h and IFN-g after 6 h. The levels of IL-2 and IFN-g in CD8 þ and CD8 ¹ T cells in four healthy individuals were consistent on four occasions over a 3-month period. In a large group of 34 normal subjects, there was considerable heterogeneity in CD3/IL-2 þ (range 9 . 7-41 . 3) and CD3/IFN-g þ cells (10 . 1-44), expressed as a percentage of total lymphocytes. In patients with atopic dermatitis (n ¼ 5) there was a significantly decreased percentage of CD3 þ /CD8 þ peripheral blood T cells expressing IFN-g and an increased percentage of CD3 þ /CD8 ¹ T cells expressing IL-4 compared with non-atopic dermatitis controls (n ¼ 5). Possible applications of this technique are discussed.

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“…Similar subsets have been identified in humans [3,4], but in contrast to the mouse, production of IL-2, IL-10 and IL-13 is shared by both classes of Th cells.…”

Section: Introductionsupporting

confidence: 62%

Intracellular cytokine expression in whole blood preparations from normals and patients with atopic dermatitis

Ferry

Antrobus

Hužička

et al. 1997

Clinical and Experimental Immunology

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“…16 Mucosal specimens were disrupted and single T-cell blasts were cloned under limiting dilution (0.3 cells/well) as reported previously. 16,17 Each clone was screened (in triplicate cultures for each condition) for responsiveness to H. pylori antigens by measuring [ 3 H]thymidine uptake after 60 hours' stimulation with medium, H. pylori lysate (aqueous extract of NCTC11637 strain, 10 µg/mL being optimal), purified urease, 18 recombinant cytotoxin-associated protein (CagA), recombinant vacuolating cytotoxin (VacA), or recombinant heat shock protein (HSP) (1 µg/mL was optimal) in the presence of irradiated autologous mononuclear cells as antigenpresenting cells (APCs), as previously reported in detail. 16 A mitogenic index greater than 10 was considered a positive result.…”

Section: Generation Of H Pylori-specific T-cell Clonesmentioning

confidence: 99%

“…T-cell clones specific for purified protein derivative (PPD) of Mycobacterium tuberculosis were derived from peripheral blood T cells of all patients, as reported. 17 …”

Section: Generation Of H Pylori-specific T-cell Clonesmentioning

confidence: 99%

“…16 To induce cytokine production by gastric T-cell clones with unknown specificity, T-cell blasts were stimulated for 36 hours with phorbol-12-myristate 13-acetate (PMA, 10 ng/mL) plus anti-CD3 monoclonal antibody (200 ng/mL), as detailed previously. 17 Duplicate samples of each supernatant were assayed for interferon (IFN)-␥ (BioSource International, Camarillo, CA), IL-4, and IL-5 (R&D Systems, Minneapolis, MN). IL-2, IFN-␥, IL-4, IL-5, and IL-13 (and L32 or GAPDH house-keeping genes, as controls) messenger RNA expression was determined by a multiprobe RNAse Protection Assay system (Pharmingen, San Diego, CA), according to the manufacturer's protocol.…”

Section: Cytokine Profile Of H Pylori-specific Gastric T-cell Clonesmentioning

confidence: 99%

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Impaired T-cell regulation of B-cell growth in Helicobacter pylori–related gastric low-grade MALT lymphoma

D’Elios¹,

Amedei²,

Manghetti³

et al. 1999

Gastroenterology

101

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“…Production of IL-2, IL-4, IL-5 and interferon-gamma (IFN-g) allows one to identify two discrete subsets of CD4 þ T cells, namely T helper (Th)1 and Th2, respectively [11][12][13][14]. Th1 cells produce IL-2 and IFN-g, but little or no IL-4 or IL-5.…”

Section: Introductionmentioning

confidence: 99%

Synovial fluid T cell clones from oligoarticular juvenile arthritis patients display a prevalent Th1/Th0-type pattern of cytokine secretion irrespective of immunophenotype

Gattorno¹,

Facchetti

Ghiotto

et al. 1997

Clinical and Experimental Immunology

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SUMMARYThe aim of the present study was to investigate the patterns of cytokine production by T cell clones raised from in vivo activated synovial fluid (SF) mononuclear cells (MNC) of five patients with oligoarticular juvenile arthritis (JA). Freshly isolated SF T cells were cultured in vitro with low dose recombinant IL-2 and subsequently cloned by limiting dilution. Sixty-four clones were obtained from the five patients studied. Fifty-nine clones were TCR a/b þ , either CD4 þ (n ¼ 43) or CD8 þ (n ¼ 15). The remaining five clones were TCR g/d þ , CD4 ¹ , CD8 ¹ . Clone immunophenotypes differed in the individual patients. Forty-four T cell clones were stimulated with phytohaemagglutinin (PHA) and phorbol myristate acetate (PMA) and supernatants tested for the presence of IL-2, IL-4, IL-5 and interferon-gamma (IFN-g) by ELISA or bioassays. Cytokine mRNA accumulation was tested by reverse transcriptase-polymerase chain reaction (RT-PCR). Most of 44 clones tested released large amounts of IFN-g irrespective of the immunophenotype. Of these, 27 were classified as Th1-type and 17 as Th0-type based upon the IFN-g/IL-4 ratio in culture supernatants. Finally, when 10 representative T cell clones were tested for pro-and anti-inflammatory cytokines, gene expression by RT-PCR, all of them were found to express the granulocyte-macrophage colony-stimulating factor (GM-CSF), tumour necrosis factor-alpha (TNF-a), IL-10 and transforming growth factor-beta 1 (TGF-b1) genes, and half of them IL-6 and IL-8 mRNA. In conclusion, T cell clones, that represent the progeny of in vivo activated SF T cells from oligoarticular JA patients, display heterogeneous immunophenotypes, but all share the ability to produce large amounts of IFN-g, with a predominant Th1/Th0 pattern. The expression of pro-and anti-inflammatory cytokine genes in these clones suggests that in vivo activated SF T cells modulate joint inflammation in a complex fashion.

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Purified protein derivative of Mycobacterium tuberculosis and excretory-secretory antigen(s) of Toxocara canis expand in vitro human T cells with stable and opposite (type 1 T helper or type 2 T helper) profile of cytokine production.

Cited by 654 publications

References 22 publications

Intracellular cytokine expression in whole blood preparations from normals and patients with atopic dermatitis

Intracellular cytokine expression in whole blood preparations from normals and patients with atopic dermatitis

Impaired T-cell regulation of B-cell growth in Helicobacter pylori–related gastric low-grade MALT lymphoma

Synovial fluid T cell clones from oligoarticular juvenile arthritis patients display a prevalent Th1/Th0-type pattern of cytokine secretion irrespective of immunophenotype

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