1. Flexible migration program regulates γδ T-cell involvement in humoral immunity
by Marlène Brandes, Katharina Willimann, Alois B. Lang, Ki-Hoan Nam, Chenggang Jin, Michael B. Brenner, Craig T. Morita, and Bernhard Moser
Blood
Volume 102(10):
November 15, 2003
©2003 by American Society of Hematology

2. nflammatory migration program in human peripheral blood-derived γδ T-cell lines.
nflammatory migration program in human peripheral blood-derived γδ T-cell lines. (A) Flow cytometric analysis of chemokine receptors (open histograms) on resting (previously activated, nonproliferating), IPP-selective γδ T-cell lines derived from human peripheral blood. Filled histograms represent control staining with isotype-matched Abs, and numbers refer to the percent receptor-positive cells. (B) Chemotactic migration and (C) Ca2+ mobilization responses in resting IPP-selective γδ T cells. Migration data, expressed as the fraction (percent) of input cells that have migrated (± SD), correspond to maximal responses determined by titration of chemokine concentrations; BCA-1 was only active at the highest concentration (3 μM) tested. Maximal chemotaxis responses were obtained with 1 or 10 nM I-TAC, SDF-1, MCP-1, RANTES, MDC, and MIP-1β; 100 nM eotaxin and LARC; 1 μM SLC; and 3 μM BCA-1. Ca2+ mobilization data correspond to 100 nM of chemokines. Functional data are representative of at least 3 independent experiments.
Marlène Brandes et al. Blood 2003;102:
©2003 by American Society of Hematology

3. Stimulation of cultured γδ T cells rapidly induces an LN-homing program.
Stimulation of cultured γδ T cells rapidly induces an LN-homing program. (A) Change in chemokine receptor expression during stimulation of peripheral blood-derived γδ T-cell lines with IPP. Positivity for CCR7 (□), CCR4 (▵), and CXCR4 (○) was determined by flow cytometry as in Figure 1 and is expressed as the fraction (percent) of receptor-positive cells. (B) Chemotactic migration profile (±SD) of activated γδ T cells after 3 days of stimulation with IPP was examined as described in Figure 1. Asterisk points out that the migration response to LARC was examined with γδ T cells after 6 days instead of 3 days of stimulation, reflecting a delay in maximal CCR6 expression. One of 3 independent experiments is shown.
Marlène Brandes et al. Blood 2003;102:
©2003 by American Society of Hematology

4. γδ T cells and αβ T cells in peripheral blood differ fundamentally in their respective chemokine receptor expression profiles.
γδ T cells and αβ T cells in peripheral blood differ fundamentally in their respective chemokine receptor expression profiles. (A) Dot plot diagrams show double staining data for CD45RO and chemokine receptors in freshly isolated peripheral blood γδ T cells (upper row) and αβ T cells (lower row). The data are representative of a study that included peripheral blood from a total of 19 donors. (B) γδ T cells and αβ T cells from the same donor blood differ substantially in their levels of cell surface CCR7 and CXCR5. Chemokine receptor staining data (open histograms) are plotted over control stainings with isotype-matched Abs (filled histograms). Bars define gates for receptor-positive cells, and numbers refer to the percent of receptor-positive cells. The Abs used in flow cytometric analysis are listed in parentheses and include 1 rat mAb (3D12) and 2 mouse mAbs (2H12, ) against CCR7 and 1 mouse mAb ( ) against CXCR5. The data are representative of a comparative analysis with peripheral blood T cells from 3 healthy individuals.
Marlène Brandes et al. Blood 2003;102:
©2003 by American Society of Hematology

5. Rapid switch in the migration program during activation of freshly isolated peripheral blood γδ T cells.
Rapid switch in the migration program during activation of freshly isolated peripheral blood γδ T cells. (A) Chemokine receptor expression is shown in freshly isolated peripheral blood γδ T cells (dashed lines) and activated γδ T cells after 36 hours of stimulation with IPP (bold lines). The control isotype Ab stainings (filled histograms) are shown for activated γδ T cells. Note the inverse regulation of CCR5 and CCR7 expression during γδ T-cell activation. (B) Chemotactic migration profile of activated peripheral blood γδ T cells after 36 hours of stimulation with IPP was examined as described in Figure 1. Maximal migration responses (±SD) were observed at 1 or 10 nM I-TAC, SDF-1, RANTES, and MDC; 1 μM SLC; and 3 μM BCA-1. One of 3 independent experiments is shown.
Marlène Brandes et al. Blood 2003;102:
©2003 by American Society of Hematology

6. γδ T cells cluster in B-cell follicles within secondary lymphoid tissues.
γδ T cells cluster in B-cell follicles within secondary lymphoid tissues. Immunohistochemical analysis of tissue section from human biopsy specimen by means of bright field (A-C) and fluorescence microscopy (D-E). (A) Normal abdominal LN, (B) gut follicle from a patient with ileitis (Yersinia), and (C) draining LN from a patient with appendicitis (Yersinia). Red staining in panels A-C depicts positivity for γδ T cells using a goat Ab against pan-VγVδ-TCR or BCA-1 and SLC as indicated using mouse mAb, and IgG refers to negative control staining with isotype-matched unrelated Ab. (D-E) Assessment of γδ T-cell distribution in sections of an abdominal LN from a patient with yersiniosis. Double-fluorescence images show localization of γδ T cells in green, CD3+ T cells (D) and CD21+ FDCs (E) in red, and the corresponding superimposed images (overlays).
Marlène Brandes et al. Blood 2003;102:
©2003 by American Society of Hematology

7. γδ T cells provide potent B-cell help.
γδ T cells provide potent B-cell help. (A) Activation-dependent expression of costimulatory molecules in γδ T cells is delayed but sustained. Flow cytometric analysis of costimulatory molecules on peripheral blood γδ T cells that were analyzed either immediately after isolation (top row) or after 36 hours (middle row) and 84 hours (lower row) of stimulation with IPP. Culturing of γδ T cells in the absence of TCR triggering (but in the presence of IL-2) did not result in marked expression of costimulatory molecules, as evidenced in cells at 84 hours of culture (dashed line). Control Ab stainings are shown in filled histograms. (B) γδ T cells induce Ab production during coculture with B cells. Tonsillar B cells were cultured alone (-)orin the presence of CXCR5-negative αβ T cells (TC), TFH cells (TFH), or peripheral blood γδ T cells (γδ T) without addition of exogenous cytokines; after 10 days the Ab titers in the culture supernatants were determined by ELISA. One of 4 representative experiments is shown.
Marlène Brandes et al. Blood 2003;102:
©2003 by American Society of Hematology