In vitro growth and phenotypic characterization of mesodermal-derived and epithelial components of normal and abnormal human thymus

Singer, Kay H.; Harden, Elizabeth; Robertson, Annette L.; Lobach, David F.; Haynes, Barton F.

doi:10.1016/0198-8859(85)90009-6

Cited by 67 publications

(39 citation statements)

References 30 publications

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“…Where these thymocytes were found in contact with epithelial cell processes, Ti molecules were localized in the region of thymocyte-epithelial contact. Recent development of methods for the long-term culture of human thymic-stromal elements (11), as well as development of monoclonal reagents specific for nonlymphoid components of human thymus (12)(13)(14)(15), have made it possible to investigate thymocyte-stromal interactions in vitro. In this report we present evidence that human thymocytes bind to both autologous and allogeneic thymic epithelial (TE) (16).…”

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confidence: 99%

“…Human peripheral blood lymphocytes and tonsillar lymphocytes were prepared as described (16), and T lymphocytes were isolated by filtration over nylon wool. TE cell cultures were initiated, propagated, and subcultured as described (11). Cytocentrifuge preparations of cultured TE cells were evaluated in indirect immunofluorescence assays using a panel of monoclonal antibodies including AE-1 (anti-keratin) (17), Mo-1 (anti-monocyte, macrophage) (18), Leu-M3 (anti-monocyte, macrophage) (19), (anti-fibroblast) (13), and L243 (anti-MHC class II antigen) (20).…”

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Human thymocytes bind to autologous and allogeneic thymic epithelial cells in vitro.

Singer¹,

Wolf²,

Lobach³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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The thymus plays a critical role in the generation of immunocompetent T lymphocytes. In the thymus, lymphocytes are in close contact with epithelial cells, and this contact is necessary for T-cell maturation. Using cultured human thymic epithelial (TE) cells, we have found that human thymocytes bind to human TE cells in vitro. Thymocytes bound to both allogeneic and autologous TE cells and to the epidermoid carcinoma cell line A431 but did not bind to epidermal keratinocytes or to thymic fibroblasts. Thymocyte binding to TE cells was trypsin-and cytochalasin B-sensitive. Indirect immunofluorescence assays showed that both mature (T6-, T3+) and immature (T6+, T3-) thymocytes bound TE cells. In our system, TE-thymocyte binding was not inhibited by antibodies to class I or class II major histocompatibility antigens. In vitro binding of thymocytes to TE cells may represent a correlate of in vivo TE-thymocyte interactions and provides a model system for the study of human intrathymic T-lymphocyte maturation and activation.Microscopists have long appreciated the extensive physical contact between the lymphoid and nonlymphoid elements of the thymus (1, 2). Direct contact of lymphoid and nonlymphoid elements within the thymus is necessary for generation of functionally mature, antigen-specific, major histocompatibility complex (MHC)-restricted T lymphocytes (3-6). Although the precise role played by the nonlymphoid component of the thymus is poorly understood, a number of interactions of developing thymocytes with nonlymphoid elements have been identified, including the formation of lymphoepithelial cell complexes in vivo, called thymic nurse cells (7), and the binding of thymocytes to macrophages and dendritic cells (8, 9). Farr et al. (10) recently observed a subpopulation of thymocytes within the thymic cortex expressing low levels of surface T-lymphocyte antigen receptors (Ti). Where these thymocytes were found in contact with epithelial cell processes, Ti molecules were localized in the region of thymocyte-epithelial contact. Recent development of methods for the long-term culture of human thymic-stromal elements (11), as well as development of monoclonal reagents specific for nonlymphoid components of human thymus (12-15), have made it possible to investigate thymocyte-stromal interactions in vitro. In this report we present evidence that human thymocytes bind to both autologous and allogeneic thymic epithelial (TE) tTo whom reprint requests should be addressed. 6588The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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mentioning

confidence: 99%

mentioning

confidence: 99%

Human thymocytes bind to autologous and allogeneic thymic epithelial cells in vitro.

Singer¹,

Wolf²,

Lobach³

et al. 1986

Proc. Natl. Acad. Sci. U.S.A.

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“…Fibroblast-like synovial cells were grown in an explant culture system using techniques previously described for growing thymic epithelium (20). Briefly, synovium was cut into l-mm3 pieces, placed under a coverslip anchored with sterile vacuum grease, and cultured while submerged in enriched media.…”

Section: Methodsmentioning

confidence: 99%

Synovial microenvironment‐t cell interactions

Haynes

Grover

Whichard

et al. 1988

Arthritis & Rheumatism

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Synovitis in rheumatoid arthritis is characterized by infiltration of the synovium by T and B lymphocytes and monocytes, as well as by the proliferation of synovial lining cells, fibroblasts, and endothelial cells. To study synovial cell‐T cell interactions in vitro, we established cultures of fibroblast‐like synovial cells, and used these cells in a synovial cell‐T cell binding assay. Using T cells at various stages of differentiation and activation, we found that human thymocytes and mitogen‐activated peripheral blood T cells bound to fibroblast‐like synovial cells, whereas fresh peripheral blood T cells did not. Moreover, activated T cells from inflammatory synovial tissue or from synovial fluid also bound to fibroblast‐like synovial cells cultured in vivo. Antibodies against certain epitopes of the T cell CD2 (35.1) and synovial cell lymphocyte function‐associated antigen‐3 (LFA‐3) (TS2/9) molecules inhibited synovial cell‐thymocyte binding. However, these same anti‐CD2 and anti‐LFA‐3 antibodies only partially inhibited synovial cell binding to activated normal peripheral blood T cells. Moreover, T cells from inflammatory synovium from rheumatoid arthritis and psoriatic arthritis patients also bound to synovial cells in vitro. These findings demonstrate that fibroblast‐like synovial cells are capable of binding to human T cells in vitro, and suggest that during the course of inflammatory synovitis, synovial fibroblast‐T cell interactions may occur in vivo.

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“…CTEF cultures were established by an explant technique and subcultured as previously described [22][23][24][25][26]. Three to five grams of thymus were obtained, the thymic capsule removed, and the tissue minced into one mm fragments and agitated gently in Dulbecco's modified Eagle's medium (DMEM) (GIBCO, BRL, Grand Island, NY) supplemented with 5% fetal calf serum (FCS) to wash out as many thymocytes as possible.…”

Section: Ctefsmentioning

confidence: 99%

“…Three to five grams of thymus were obtained, the thymic capsule removed, and the tissue minced into one mm fragments and agitated gently in Dulbecco's modified Eagle's medium (DMEM) (GIBCO, BRL, Grand Island, NY) supplemented with 5% fetal calf serum (FCS) to wash out as many thymocytes as possible. Thymocytes and other hematopoietic cells, including dendritic cells, were depleted by incubation in 1.35 mM 2′-deoxyguanosine (Sigma, St. Louis, MO) [22]. The CTEF were incubated on sterile gelfoam sponges, 1 cm × 3 cm, in 6 cm Petri dishes in 12 ml of 1.35 mM 2′-deoxyguanosine and Ham's F-12 medium supplemented with 10% FCS, 25 mM HEPES, 2 mM glutamine, 50 U/ml penicillin, 1 µg/ml gentamicin, 50 µg/ml streptomycin and 2 µg/ml amphotericin (complete medium) at 37˚C in a 5% CO 2 atmosphere for two weeks.…”

Section: Ctefsmentioning

confidence: 99%

T Cell Differentiation/Maturation of CD34⁺Stem Cells from HIV‐Seropositive Hemophiliacs in Cultured Thymic Epithelial Fragments

et al. 1996

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In summary, these studies demonstrated that HIV-infected hemophiliac bone marrowderived nonadherent CD34 + SC were capable of differentiating and/or maturing into T cells when cocultured in a normal allogeneic thymic environment. Furthermore, the T cells derived from derived CD34 + SC were capable of differentiating into T cells when cocultured in a normal allogeneic thymic environment, proliferated maximally with APCs from the stem cell donor and were tolerant of thymic HLA class II antigens, and to a lesser degree to stem cell donor B cell HLA antigens.

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In vitro growth and phenotypic characterization of mesodermal-derived and epithelial components of normal and abnormal human thymus

Cited by 67 publications

References 30 publications

Human thymocytes bind to autologous and allogeneic thymic epithelial cells in vitro.

Human thymocytes bind to autologous and allogeneic thymic epithelial cells in vitro.

Synovial microenvironment‐t cell interactions

T Cell Differentiation/Maturation of CD34⁺Stem Cells from HIV‐Seropositive Hemophiliacs in Cultured Thymic Epithelial Fragments

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In vitro growth and phenotypic characterization of mesodermal-derived and epithelial components of normal and abnormal human thymus

Cited by 67 publications

References 30 publications

Human thymocytes bind to autologous and allogeneic thymic epithelial cells in vitro.

Human thymocytes bind to autologous and allogeneic thymic epithelial cells in vitro.

Synovial microenvironment‐t cell interactions

T Cell Differentiation/Maturation of CD34+Stem Cells from HIV‐Seropositive Hemophiliacs in Cultured Thymic Epithelial Fragments

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T Cell Differentiation/Maturation of CD34⁺Stem Cells from HIV‐Seropositive Hemophiliacs in Cultured Thymic Epithelial Fragments