Cell Biology of Cytotoxic and Helper T Cell Functions: Immunofluorescence Microscopic Studies of Single Cells and Cell Couples

Kupfer, Abraham; Singer, S. J.

doi:10.1146/annurev.iy.07.040189.001521

Cited by 279 publications

(122 citation statements)

References 63 publications

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“…:ales which are guided by microtubules to release their contents ~nly towards the site of target cell contact [19]. The functional mportance of the FasL-Fyn interaction is unknown.…”

Section: Resultsmentioning

confidence: 99%

“…Secretory vesicles lhat have the poly-proline-rich sequence of FasL cytoplasmati-:ally exposed might be directly targeted to the cell surface via Fyn and/or Lck SH3 domains, both of which are known to cap ~t the site of TcR receptor/target cell interaction [20]. This ~mplies that the resulting polarized vesicle-cell membrane fusion could engender local exposure of other vesicle-harbored proteins, including adhesion molecules and cytokines [19]. Precipitates were analysed by immunoblotting using the respective antibodies.…”

Section: Resultsmentioning

confidence: 99%

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Interaction of peptides derived from the Fas ligand with the Fyn‐SH3 domain

Hane

Lowin

Peitsch³

et al. 1995

FEBS Letters

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Interaction of the widely expressed Fas with its membrane-hound ligand (FasL) leads to rapid cell death via apoptosis. Io avoid pathological tissue damage, the activity of FasL requires tight regulation. Here, we report that the Src homology 3 (SH3) domain of Fyn binds to the proline-rich cytoplasmic region of lZasL. Binding of the SH3 domain occurs between amino acid rt~sidues 44-71 which contains several potential SH3 interaction sites. This binding is specific, as SH3 domains of Lck, Grb2 and ras-GAP bind only weakly or not at all. We suggest that FasL activity may be modulated by SH3 domains of the src-like Fyn kinase.

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“…:ales which are guided by microtubules to release their contents ~nly towards the site of target cell contact [19]. The functional mportance of the FasL-Fyn interaction is unknown.…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Interaction of peptides derived from the Fas ligand with the Fyn‐SH3 domain

Hane

Lowin

Peitsch³

et al. 1995

FEBS Letters

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“…While the actin cytoskeleton redistributes at the leading edge, the microtubule organizing center (MTOC) localizes within the uropod allowing a greater deformability of the migrating cells (4)(5)(6). Given that the MTOC and the Golgi apparatus colocalize, the secretion of soluble mediators is directed toward the ligated cell surface (7). Moreover, chemokine receptors responsible for the directionality of the migration along a gradient of chemoattractants are concentrated within the leading edge (8).…”

mentioning

confidence: 99%

CD2 molecules redistribute to the uropod during T cell scanning: Implications for cellular activation and immune surveillance

Tibaldi

Salgia

Reinherz

2002

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

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Dynamic binding between CD2 and CD58 counter-receptors on opposing cells optimizes immune recognition through stabilization of cell-cell contact and juxtaposition of surface membranes at a distance suitable for T cell receptor-ligand interaction. Digitized time-lapse differential interference contrast and immunofluorescence microscopy on living cells now show that this binding also induces T cell polarization. Moreover, CD2 can facilitate motility of T cells along antigen-presenting cells via a movement referred to as scanning. Both activated CD4 and CD8 T cells are able to scan antigen-presenting cells surfaces in the absence of cognate antigen. Scanning is critically dependent on T cell ␤-integrin function, as well as myosin light chain kinase. More importantly, surface CD2 molecules rapidly redistribute on interaction with a cellular substratum, resulting in a 100-fold greater CD2 density in the uropod versus the leading edge. In contrast, no redistribution is observed for CD11a͞CD18 or CD45. Molecular compartmentalization of CD2, T cell receptor, and lipid rafts within the uropod prearranges the cellular activation machinery for subsequent immune recognition. This ''presynapse'' formation on primed T cells will likely facilitate the antigen-dependent recognition capability required for efficient immune surveillance.A rrival of antigen-laden dendritic cells into the lymph node results in the activation of naive T cells, leading to their proliferation and acquisition of effector functions (1). The subsequent migration of activated T cells into pathogen-invaded body tissues is a key feature of immunity and immune surveillance. Diapedesis of T cells through the vascular endothelium, as well as their crawling movement on the surface of antigen presenting cells (APCs), parenchymal cells and extracellular matrix (ECM), requires T cell polarization (2). Although the molecular basis of this process is not well understood, certain features have been identified. Cell polarization is characterized by the formation of a leading edge at the front of the cell and a uropod at its back, allowing conversion of mechanical forces generated through adhesion to a substratum into net cellular locomotion (3). While the actin cytoskeleton redistributes at the leading edge, the microtubule organizing center (MTOC) localizes within the uropod allowing a greater deformability of the migrating cells (4-6). Given that the MTOC and the Golgi apparatus colocalize, the secretion of soluble mediators is directed toward the ligated cell surface (7). Moreover, chemokine receptors responsible for the directionality of the migration along a gradient of chemoattractants are concentrated within the leading edge (8). In contrast, the uropod mediates important adhesion functions, as suggested by accumulation of a notable number of adhesion molecules including ICAM-1, CD44, CD43, P-selective glycoprotein ligand 1 (PSGL-1), and L-selectin during migration (9, 10). The uropod therefore appears to anchor the cell to the ECM, APCs, or endothelial cells w...

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“…We have described an in vitro model system to study at the single-cell level events that occur within the first minutes of the interaction of cloned Th cells and Ag-presenting B-cell (B-APC) lymphomas or hybridomas (8). By using immunofluorescence microscopy, we observed the clustering of the T-cell receptors (TCRs) for Ag, CD4, and lymphocyte function-associated antigen 1 (LFA-1) membrane receptors along with the cytoskeletal protein talin into the Th-B contact area and the reorientation of the microtubule-organizing center (MTOC) inside the Th cell to face the bound B-APC (9)(10)(11)(12).…”

mentioning

confidence: 99%

Polarized expression of cytokines in cell conjugates of helper T cells and splenic B cells.

Kupfer

Mosmann

Kupfer

1991

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

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218

154

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We describe the intracellular localization, by double immunofluorescence microscopy, of four cytokines that were produced during the prolonged interaction of cloned helper T cells with resting splenic B cells. When two rabbit immunoglobulin-specific helper-T-cell clones were mixed, either separately or together, with splenic B cells in the presence of the antigen rabbit anti-mouse immunoglobulin antibodies, stable T-cell-B-cell conjugates were seen up to 29 hr later.Microscopic observations of these cells revealed that interferon y and interleukin 2, inside one of the T-cell clones, and interleukins 4 and 5, inside the other T-cell clone, were concentrated very close to the T-cell-B-cell contact area. The cytokines were not seen in the T cells prior to their interaction with the B cells and their production was strictly antigenspecific. These studies show, at the single-cell level, that helper-T-cell clones can remain bound to splenic B cells long enough for the T cells to produce cytokines, which are synthesized near the bound B cells. We propose that the polarized synthesis of the cytokines may result in their directed secretion toward the bound B cells. By locally secreting the cytokines, which are not antigen-specific, at the contacting T-cell-B-cell membranes, where T-and B-cell surface receptors are engaged and clustered, the helper T cells can induce selective and specific B-cell responses.The introduction of proteinaceous foreign antigens (Ags) into an animal generally results in the proliferation and differentiation of B cells that then secrete antibodies (Abs), which bind specifically to the foreign Ags. The B-cell response is largely controlled by soluble growth and differentiation factors, collectively termed lymphokines or cytokines (1, 2), which are produced by Ag-specific helper T cells (Th cells).It is unclear yet how selective and regulated B-cell responses can be generated by the secretion of cytokines such as interferon y (IFN-y) and interleukins 2, 4, and 5 (IL-2, IL-4, and IL-5, respectively), since these cytokines are not Agspecific or even cell-type-specific (2). In addition, each of these cytokines can have multiple effects on the cells and when present together they can either antagonize or synergize each other (3). However, the addition of crude or purified cytokines to resting B cells in the absence of Th cells failed to cause B-cell proliferation and differentiation, implying that specific cell-cell interactions may be additionally required for effective B-cell responses (4-7). The molecular events that take place during such Th-cell-B-cell (Th-B) interactions are still largely unknown.We have described an in vitro model system to study at the single-cell level events that occur within the first minutes of the interaction of cloned Th cells and Ag-presenting B-cell (B-APC) lymphomas or hybridomas (8). By using immunofluorescence microscopy, we observed the clustering of the T-cell receptors (TCRs) for Ag, CD4, and lymphocyte function-associated antigen 1 (LFA-1) membrane re...

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Cell Biology of Cytotoxic and Helper T Cell Functions: Immunofluorescence Microscopic Studies of Single Cells and Cell Couples

Cited by 279 publications

References 63 publications

Interaction of peptides derived from the Fas ligand with the Fyn‐SH3 domain

Interaction of peptides derived from the Fas ligand with the Fyn‐SH3 domain

CD2 molecules redistribute to the uropod during T cell scanning: Implications for cellular activation and immune surveillance

Polarized expression of cytokines in cell conjugates of helper T cells and splenic B cells.

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