Korean J Physiol Pharmacol.  2012 Aug;16(4):243-247. 10.4196/kjpp.2012.16.4.243.

Morphologic Evidence of Anti-Tumor Specificity of T Cells Activated by Denritic Cells Derived from Peripheral Blood Mononuclear Cells of Thyroid Cancer Patients

• Lee DH

• KMID: 2011158
• DOI: http://doi.org/10.4196/kjpp.2012.16.4.243

Abstract

Recent studies suggest that immunization with autologous dendritic cells (DCs) results in protective immunity and rejection of established tumors in various human malignancies. The purpose of this study is to determine whether DCs are generated from peripheral blood mononuclear cells (PBMNs) by using cytokines such as F1t-3 ligand (FL), granulocyte macrophage-colony stimulating factor (GM-CSF), IL-4, and TNF-alpha, and whether cytotoxic T cells activated against the thyroid cancer tissues by the DCs. Peripheral blood was obtained from 2 patients with thyroid cancer. DCs were established from PBMNs by culturing in the presence of FL, GM-CSF, IL-4, and TNF-alpha for 14 days. At day 14, the differentiated DCs was analyzed morphologically. The immunophenotypic features of DCs such as CDla, CD83, and CD86 were analyzed by immunofluorelescence microscopy. At day 18, DCs and T cells were incubated with thyroid cancer tissues or normal thyroid tissues for additional 4 days, respectively. DCs generated from the PBMNs showed the typical morphology of DCs. Activated cytotoxic T lymphocytes (CTLs) were observed also. DCs and the CTLs were attached to the cancer tissues on scanning electron microscope. The DCs activated the CTLs, which able to specifically attack the thyroid cancer. This study provides morphologic evidence that the coculture of T cells/cancer tissues activated the T cells and differentiated CTLs. The CTLs tightly adhered to cancer tissues and lysed cancer tissues vigorously. Therefore DCs could be used as potential vaccines in the immunotherapy.

Keyword

Anti-tumor specificity; Cytotoxic T lymphocytes (CTLs); Dendritic cells; Immunotherapy; Thyroid cancer

MeSH Terms

Coculture Techniques
Cytokines
Dendritic Cells
Electrons
Granulocyte-Macrophage Colony-Stimulating Factor
Granulocytes
Humans
Immunization
Immunotherapy
Interleukin-4
Microscopy
Rejection (Psychology)
Sensitivity and Specificity*
T-Lymphocytes
T-Lymphocytes, Cytotoxic
Thyroid Gland
Thyroid Neoplasms
Tumor Necrosis Factor-alpha
Vaccines
Cytokines
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-4
Tumor Necrosis Factor-alpha
Vaccines

Figure

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  Fig. 1 Phase contrast micrograph of freshly isolated PBMNs showing a homogenous population of equally sized and round cells. Original magnification ×400 (A). After 5 days of culture with medium containing GM-CSF, IL-4, TNF-α and FL, spindle-shape cells showed in the center. Original magnification ×400 (B). After 8 days of culture with medium containing GM-CSF, IL-4, TNF-α and FL, the cell with large-cell bodies and long dendritic projections was visible. Original magnification ×400 (C).

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  Fig. 2 The cells with long cytoplasmic projections were visible at 14 days of culture in medium containing GM-CSF, IL-4, TNF-α and FL. Original magnification ×400.

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  Fig. 3 The microscopic finding of non-neoplastic thyroid tissue culture after being formalin fixed paraffin embedded. This view shows dilated thyroid follicles. H-E stain. Original magnification ×100.

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  Fig. 4 Round to oval cells are arranged in clusters, and acinar pattern with intersecting scanty amyloid deposits. Tumor cells contains uniform, round to oval nuclei with finely granular chromatin, and small and visible nucleoli. Cytoplasm is abundant eosinophilic. H-E stain. Original magnification ×400.

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  Fig. 5 Scanning electron micrograph of a veiled cell. A veiled cell is attached to the papillary thyroid cancer tissue. Original magnification ×5,000.

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  Fig. 6 Scanning electron micrograph of a non-activated T cell. This view shows a T cell cultured with normal thyroid tissue. Original magnification ×5,000.

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  Fig. 7 Scanning electron micrograph of activated T cell. An activated T cell adheres to the papillary thyroid cancer tissue and attacks the carcinoma tissue. Original magnification ×5,000.

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  Fig. 8 Fluorescence micrograph of cultured DCs showing intense staining with the monoclonal antibodies for specific cell surface immunophenotype. DCs showed the positive FITC staining. (A) CD1a, (B) CD83, (C) CD86.

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