The tumor suppressor Adenomatous polyposis coli modulates T lymphocyte migration

Adenomatous polyposis coli (APC) is a tumor suppressor whose mutations underlie familial adenomatous polyposis (FAP) and colorectal cancer. Although its role in intestinal epithelial cells is well characterized, APC importance for anti-tumor immunity is ill defined. Besides its role in Wnt/β-catenin signaling, APC regulates cytoskeleton organization, cell polarity and migration in various cells types. Here we address whether APC plays a role in T lymphocyte migration, a key step of anti-tumor immune responses. Using a series of cell biology tools, we measured migration of primary T cells obtained from FAP patients carrying APC mutations. FAP T cells showed decreased chemotaxis through micropores or endothelial cell monolayers. Concomitantly, they presented lower expression of the VLA-4 (α4β1) integrin at the cell surface. Notably, adhesion and migration in micro- fabricated channels were specifically reduced when surfaces were coated with VLA-4 ligands, indicating that defective adhesion could lead to decreased T cell migration. To further dissect the cellular mechanisms underpinning APC-mediated defects, we depleted APC in the CEM T cell line. We found that APC is critical for VLA-4-dependent adhesion, and acto-myosin and microtubule organization in migrating cells. APC-silenced CEM cells preferentially adopt an ameboid-like migration, lacking adhesive filopodia and continuously extending and retracting unstructured membrane protrusions. These findings underscore a role of APC in T cell migration via modulation of integrin- dependent adhesion and cytoskeleton reorganization. Hence, APC mutations in FAP patients not only unbalance epithelial homeostasis, driving intestinal neoplasms, but also impair T cell migration, potentially leading to inefficient T cell-mediated anti-tumor immunity.

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A Novel Role for CpG Oligonucleotides in Tumor Immunotherapy: CpG-ODN Induce Targeted Chemokine-Induced Lymphocyte Migration to the Peripheral Tissues in Humans.

Blood ◽

10.1182/blood.v110.11.1791.1791 ◽

2007 ◽

Vol 110 (11) ◽

pp. 1791-1791

Author(s):

W. Nicholas Haining ◽

Holger Kanzler ◽

Jeffrey Davies ◽

Linda Drury ◽

Jeffrey Kutok ◽

...

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Vaccine Adjuvant ◽

Cpg Odn ◽

Lymphocyte Migration ◽

Peripheral Tissues ◽

Gm Csf ◽

T Cell Migration ◽

Cell Counts

Abstract CpG ODN are being actively investigated as cancer vaccine adjuvants because they mature plasmacytoid dendritic cells (pDC) into potent antigen-presenting cells. In addition, TLR ligands induce a broad range of other immunologic effects in pDC including the secretion of interferon α (IFNa) and chemokines which alter lymphocyte migration. Whether these CpG-ODN driven TLR ligand signals enhance antigen specific Immunity and/or trafficking In humans Is presently unknown. We evaluated the efficacy of the CpG-ODN, 1018-ISS, as a vaccine adjuvant given with GM-CSF to induce T cell immunity in humans to the tumor antigen hTERT. Seventeen patients with advanced solid tumors were treated with 6 cycles of GM-CSF (x 3d), CpG-ODN (escalating from 3mg - 100mg × 1d) followed by a peptide vaccine (a CD8 epitope from hTERT), in a Phase I dose escalation study. Surprisingly, only one of seventeen patients showed a detectable hTERT-specific tetramer T cell response. However, the majority of patients developed marked peripheral blood (PB) lymphopenia after CpG-ODN. Time-course flow cytometry analysis of PB revealed that CD8, CD4, NK and B cell counts were all depressed immediately after CpG-ODN. The effect was transient, with normal counts returning after a week, suggesting that CpG-ODN induced alteration in cell migration rather than cell death. To find further evidence for altered migration we examined vaccine sites. Clinically, vaccine sites showed significant swelling/induration within hours of CpG-ODN administration, though none was dose-limiting. Immunohistochemistry of vaccine biopsies showed significant, perivascular accumulation of CD4 and CD8 T cells clustered around CD123+ pDC. Biopsies after CpG-ODN, but not after GM-CSF, showed a marked increase in expression of MxA, an interferon-inducible gene suggesting that the local activation of pDC with resultant IFNa secretion. qRT-PCR confirmed significant increases in a panel of IFNa-inducible genes in the PB after CpG-ODN, indicating a systemic effect of IFNa secretion. Lastly, we showed directly that CpG-ODN markedly increased the ability of purified pDC to induce T cell migration in an in vitro transwell assay, demonstrating that CpG-ODN stimulation of human pDC not only induces IFNa, but also other chemokines that are sufficient to chemoattract T cells. Our results show that CpG-ODN with GM-CSF may not be an effective adjuvant strategy for peptide tumor vaccines; but sequenced GM-CSF/CpG-ODN causes a chemokine response that effects T cell migration to the peripheral tissues. These findings suggest a role for CpG beyond that of a vaccine adjuvant as a mediator of lymphocyte migration, targeting immune responses to specific peripheral tissues. Therapeutic intratumoral GM-CSF/CpG-ODN injection could profoundly alter the local immunologic milieu, recruiting activated pDC and T cells to the tumor site, and tipping the balance towards an effective tumor-specific immune response.

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Protein malnutrition promotes dysregulation of molecules involved in T cell migration in the thymus of mice infected with Leishmania infantum

Scientific Reports ◽

10.1038/srep45991 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 14

Author(s):

Monica Losada-Barragán ◽

Adriana Umaña-Pérez ◽

Sergio Cuervo-Escobar ◽

Luiz Ricardo Berbert ◽

Renato Porrozzi ◽

...

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Leishmania Infantum ◽

Ex Vivo ◽

Protein Malnutrition ◽

Lymphocyte Migration ◽

Protein Levels ◽

T Cell Migration ◽

Chemotactic Factors

Abstract Protein malnutrition, the most deleterious cause of malnutrition in developing countries, has been considered a primary risk factor for the development of clinical visceral leishmaniasis (VL). Protein malnutrition and infection with Leishmania infantum leads to lymphoid tissue disorganization, including changes in cellularity and lymphocyte subpopulations in the thymus and spleen. Here we report that protein malnutrition modifies thymic chemotactic factors by diminishing the CCL5, CXCL12, IGF1, CXCL9 and CXCL10 protein levels in infected animals. Nevertheless, T cells preserve their migratory capability, as they were able to migrate ex vivo in response to chemotactic stimuli, indicating that malnutrition may compromise the thymic microenvironment and alter in vivo thymocyte migration. Decrease in chemotactic factors protein levels was accompanied by an early increase in the parasite load of the spleen. These results suggest that the precondition of malnutrition is affecting the cell-mediated immune response to L. infantum by altering T cell migration and interfering with the capacity of protein-deprived animals to control parasite spreading and proliferation. Our data provide evidence for a disturbance of T lymphocyte migration involving both central and peripheral T-cells, which likely contribute to the pathophysiology of VL that occurs in malnourished individuals.

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Activation of Wnt5A signaling is required for CXC chemokine ligand 12–mediated T-cell migration

Blood ◽

10.1182/blood-2008-08-175869 ◽

2009 ◽

Vol 114 (7) ◽

pp. 1366-1373 ◽

Cited By ~ 41

Author(s):

Manik C. Ghosh ◽

Gary D. Collins ◽

Bolormaa Vandanmagsar ◽

Kalpesh Patel ◽

Margaret Brill ◽

...

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Cxcr4 Signaling ◽

T Cell Migration ◽

Wnt5a Expression ◽

Cxc Chemokine ◽

Chemokine Ligand ◽

And Migration

Abstract Chemokines mediate the signaling and migration of T cells, but little is known about the transcriptional events involved therein. Microarray analysis of CXC chemokine ligand (CXCL) 12−treated T cells revealed that Wnt ligands are significantly up-regulated during CXCL12 treatment. Real-time polymerase chain reaction and Western blot analysis confirmed that the expression of noncanonical Wnt pathway members (eg, Wnt5A) was specifically up-regulated during CXCL12 stimulation, whereas β-catenin and canonical Wnt family members were selectively down-regulated. Wnt5A augmented signaling through the CXCL12-CXCR4 axis via the activation of protein kinase C. Moreover, Wnt5A expression was required for CXCL12–mediated T-cell migration, and rWnt5A sensitized human T cells to CXCL12-induced migration. Furthermore, Wnt5A expression was also required for the sustained expression of CXCR4. These results were further supported in vivo using EL4 thymoma metastasis as a model of T-cell migration. Together, these data demonstrate that Wnt5A is a critical mediator of CXCL12-CXCR4 signaling and migration in human and murine T cells.

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CXCL12 Enhances CLL Cell and T-Cell Migration in a Dynamic Circulating Model of CLL That Can be Abrogated By the CXCR4 Antagonist ONO-7161

Blood ◽

10.1182/blood.v124.21.3293.3293 ◽

2014 ◽

Vol 124 (21) ◽

pp. 3293-3293

Author(s):

Elisabeth J Walsby ◽

Paul Brennan ◽

Guy Pratt ◽

Tanja Nicole Hartmann ◽

Toshio Yoshizawa ◽

...

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Mononuclear Cells ◽

Cxcr4 Expression ◽

Cell Secretion ◽

Lymphocyte Migration ◽

Cxcr4 Antagonist ◽

T Cell Migration ◽

Chemokine Gradient

Abstract We have recently developed a circulating model of CLL that more accurately mimics the transient interactions that take place between lymphocytes and the vascular endothelium. Normal and malignant lymphocytes actively undergo transendothelial migration in our system in the absence of a chemoattractant. We have now refined our model to include a CXCL12 gradient and evaluated the ability of CXCR4 antagonists to modulate lymphocyte migration. Initially we demonstrated that a single deposition of CXCL12 (100ng/ml) into the extravascular space (EVS) of our model significantly increased CLL cell migration (P = 0.049, mean circulating CXCL12 level = 254pg/ml). Subsequently, we created a stable chemokine gradient by seeding CXCL12-secreting MRC5 cells into the EVS. Although this resulted in lower peak circulating levels of CXCL12 (as measured by ELISA, mean 23pg/ml), it significantly increased migration (P = 0.024) presumably because of the constant nature of the chemokine gradient. As previously shown CXCR4 expression is up-regulated on circulating CLL cells when compared to static cultures (P=0.023). Basal CXCR4 expression and expression after circulation for 48h, correlated with the degree of CLL cell migration into the EVS (r2=0.40, P=0.003; r2=0.31, P=0.009 respectively). It is worthy of note that as a proportion of all circulating peripheral blood mononuclear cells, CD3+ T-cells migrated significantly more than CD19+ CLL cells (T-cells: 12.72% ±13.47, CLL cells: 1.16% ±1.23 P =0.02, n = 10). This may be partly the result of the known defects in CLL cell migration but is also likely to be promoted by CLL cell secretion of T-cell attracting chemokines CCL3 (298pg/ml ±197.5, n=13) and CCL4 (56pg/ml ±115.6, n=13) in our model. Furthermore, migrated T-cells and CLL cells recovered from the EVS showed a marked increase in Ki-67 expression (P<0.0001 and P<0.0001 respectively) confirming that both subsets of cells are activated. We next explored whether we could block the CXCR4/CXCL12 axis using the CXCR4 antagonist ONO-7161. In transwell chambers (3μm pores) 30nM ONO-7161 resulted in significantly decreased CLL cell migration (54.1% ±25.53 of control, P=0.0003) and was more potent than Plerixafor (1μM) (P=0.06). Additionally, in the circulating system, ONO-7161 resulted in significantly reduced CLL cell migration into the EVS at 48h (P=0.0006). T-cell migration was also reduced by ONO-7161 but not significantly. In summary, we have enhanced our circulating model system of CLL by adding a physiologically relevant CXCL12 chemokine gradient. This then allowed us to dissect the role of the CXCR4/CXCL12 axis in modulating CLL cell migration. Finally we demonstrated that the CXCR4 antagonist ONO-7161 was a potent inhibitor of CLL cell migration in our model. Given that ONO-7161 was non-toxic to CLL cells at concentrations up to 10μM, it seems likely that it might be usefully combined with chemotherapy and biologically targeted agents in the treatment of this and other diseases. Disclosures Yoshizawa: Ono Pharmaceutical Co., Ltd.: Employment. Fegan:ONO PHARMA: Honoraria.

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Regulatory T cells reduce endothelial neutral sphingomyelinase 2 to prevent T cell migration into tumors

European Journal of Immunology ◽

10.1002/eji.202149208 ◽

2021 ◽

Author(s):

Paulina Akeus ◽

Louis Szeponik ◽

Veronica Langenes ◽

Viktoria Karlsson ◽

Patrik Sundström ◽

...

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Regulatory T Cells ◽

Neutral Sphingomyelinase ◽

T Cell Migration

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Novel self-amplificatory loop between T cells and tenocytes as a driver of chronicity in tendon disease

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-219335 ◽

2021 ◽

pp. annrheumdis-2020-219335

Author(s):

Emma Garcia-Melchor ◽

Giacomo Cafaro ◽

Lucy MacDonald ◽

Lindsay A N Crowe ◽

Shatakshi Sood ◽

...

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Inflammatory Cytokines ◽

T Cell Activation ◽

Cell Activation ◽

Conditioned Media ◽

Tissue Remodelling ◽

Activated T Cells ◽

T Cell Migration

ObjectivesIncreasing evidence suggests that inflammatory mechanisms play a key role in chronic tendon disease. After observing T cell signatures in human tendinopathy, we explored the interaction between T cells and tendon stromal cells or tenocytes to define their functional contribution to tissue remodelling and inflammation amplification and hence disease perpetuation.MethodsT cells were quantified and characterised in healthy and tendinopathic tissues by flow cytometry (FACS), imaging mass cytometry (IMC) and single cell RNA-seq. Tenocyte activation induced by conditioned media from primary damaged tendon or interleukin-1β was evaluated by qPCR. The role of tenocytes in regulating T cell migration was interrogated in a standard transwell membrane system. T cell activation (cell surface markers by FACS and cytokine release by ELISA) and changes in gene expression in tenocytes (qPCR) were assessed in cocultures of T cells and explanted tenocytes.ResultsSignificant quantitative differences were observed in healthy compared with tendinopathic tissues. IMC showed T cells in close proximity to tenocytes, suggesting tenocyte–T cell interactions. On activation, tenocytes upregulated inflammatory cytokines, chemokines and adhesion molecules implicated in T cell recruitment and activation. Conditioned media from activated tenocytes induced T cell migration and coculture of tenocytes with T cells resulted in reciprocal activation of T cells. In turn, these activated T cells upregulated production of inflammatory mediators in tenocytes, while increasing the pathogenic collagen 3/collagen 1 ratio.ConclusionsInteraction between T cells and tenocytes induces the expression of inflammatory cytokines/chemokines in tenocytes, alters collagen composition favouring collagen 3 and self-amplifies T cell activation via an auto-regulatory feedback loop. Selectively targeting this adaptive/stromal interface may provide novel translational strategies in the management of human tendon disorders.

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Lyp/PTPN22 is a negative regulator of integrin mediated T cell adhesion and migration; the disease associated PTPN22 allelic variant is a loss of function mutant that perturbs T cell migration

Annals of the Rheumatic Diseases ◽

10.1136/ard.2010.149096.16 ◽

2011 ◽

Vol 70 (Suppl 2) ◽

pp. A7-A7 ◽

Cited By ~ 3

Author(s):

L. Svensson ◽

G. Burn ◽

C. Sanchez-Blanco ◽

R. Zamoyska ◽

A. Cope

Keyword(s):

Cell Migration ◽

Cell Adhesion ◽

T Cell ◽

Allelic Variant ◽

Negative Regulator ◽

Loss Of Function ◽

T Cell Migration ◽

Cell Adhesion And Migration ◽

And Migration ◽

T Cell Adhesion

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Collagen Organization Does Not Influence T-Cell Distribution in Stroma of Human Pancreatic Cancer

Cancers ◽

10.3390/cancers13153648 ◽

2021 ◽

Vol 13 (15) ◽

pp. 3648

Author(s):

Eva-Maria Kamionka ◽

Baifeng Qian ◽

Wolfgang Gross ◽

Frank Bergmann ◽

Thilo Hackert ◽

...

Keyword(s):

Pancreatic Cancer ◽

T Cells ◽

Cell Migration ◽

T Cell ◽

Human Pancreatic Cancer ◽

Ductal Adenocarcinoma ◽

Cell Distribution ◽

Collagen Organization ◽

T Cell Migration ◽

Collagen Alignment

The dominant intrastromal T-cell infiltration in pancreatic cancer is mainly caused by the contact guidance through the excessive desmoplastic reaction and could represent one of the obstacles to an effective immune response in this tumor type. This study analyzed the collagen organization in normal and malignant pancreatic tissues as well as its influence on T-cell distribution in pancreatic cancer. Human pancreatic tissue was analyzed using immunofluorescence staining and multiphoton and SHG microscopy supported by multistep image processing. The influence of collagen alignment on activated T-cells was studied using 3D matrices and time-lapse microscopy. It was found that the stroma of malignant and normal pancreatic tissues was characterized by complex individual organization. T-cells were heterogeneously distributed in pancreatic cancer and there was no relationship between T-cell distribution and collagen organization. There was a difference in the angular orientation of collagen alignment in the peritumoral and tumor-cell-distant stroma regions in the pancreatic ductal adenocarcinoma tissue, but there was no correlation in the T-cell densities between these regions. The grade of collagen alignment did not influence the directionality of T-cell migration in the 3D collagen matrix. It can be concluded that differences in collagen organization do not change the spatial orientation of T-cell migration or influence stromal T-cell distribution in human pancreatic cancer. The results of the present study do not support the rationale of remodeling of stroma collagen organization for improvement of T-cell–tumor cell contact in pancreatic ductal adenocarcinoma.

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