Structure and Immune Function of Afferent Lymphatics and Their Mechanistic Contribution to Dendritic Cell and T Cell Trafficking

Afferent lymphatic vessels (LVs) mediate the transport of antigen and leukocytes to draining lymph nodes (dLNs), thereby serving as immunologic communication highways between peripheral tissues and LNs. The main cell types migrating via this route are antigen-presenting dendritic cells (DCs) and antigen-experienced T cells. While DC migration is important for maintenance of tolerance and for induction of protective immunity, T cell migration through afferent LVs contributes to immune surveillance. In recent years, great progress has been made in elucidating the mechanisms of lymphatic migration. Specifically, time-lapse imaging has revealed that, upon entry into capillaries, both DCs and T cells are not simply flushed away with the lymph flow, but actively crawl and patrol and even interact with each other in this compartment. Detachment and passive transport to the dLN only takes place once the cells have reached the downstream, contracting collecting vessel segments. In this review, we describe how the anatomy of the lymphatic network supports leukocyte trafficking and provide updated knowledge regarding the cellular and molecular mechanisms responsible for lymphatic migration of DCs and T cells. In addition, we discuss the relevance of DC and T cell migration through afferent LVs and its presumed implications on 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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Defective T Cell Migration in Chronic Lymphocytic Leukemia Is Repaired by Lenalidomide

Blood ◽

10.1182/blood.v112.11.3117.3117 ◽

2008 ◽

Vol 112 (11) ◽

pp. 3117-3117

Author(s):

Alan G. Ramsay ◽

Lena Svensson ◽

Nancy Hogg ◽

John G. Gribben

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

B Cells ◽

Chronic Lymphocytic Leukemia ◽

Cd8 T Cells ◽

Direct Contact ◽

Immune Surveillance ◽

Lymphocytic Leukemia ◽

T Cell Migration

Abstract We have previously demonstrated that multiple gene expression abnormalities are induced in T cells from chronic lymphocytic leukemia (CLL) patients including defects within the actin cytoskeleton signaling pathways that control immune recognition and motility (Gullu et al. JCI, 2005). T cell immune surveillance requires rapid migratory responses and LFA-1 (CD11a/CD18; αLβ2) is a promigratory receptor that engages the cytoskeleton to control migration. We hypothesized that CLL T cells may exhibit dysfunctional migration in response to ICAM-1, the principal ligand for LFA-1. Using time lapse microscopy, we observed significantly reduced chemokine SDF-1 (CXCL12) induced migration on ICAM-1 of CLL CD4 and CD8 T cells compared to age-matched healthy donor T cells. Healthy T cells tracked for 45 min displayed a random course of migration with an average speed of ~ 8 μm/min, whereas CLL T cells were slower ~ 5 μm/min (n=14, ~ 30% reduction, p<0.01). We further postulated that direct contact of CLL tumor cells with healthy T cells would induce this migratory defect. Healthy CD4 or CD8 T cells were cocultured with either allogeneic CLL B cells or allogeneic healthy B cells and subsequently used in migration assays. Co-culture with CLL cells resulted in significantly reduced T cell migration compared with co-culture with healthy B cells (~ 44% reduction in migration, n=6, p<0.01). Evidence that direct contact was required to induce this migratory defect was shown when no effect was observed when cell-cell adhesion was prevented by pretreatment of CLL cells with anti-ICAM-1 blocking antibody prior to primary co-culture with healthy T cells. This cancer-induced migratory defect was repaired when CLL T cells were pretreated with the immunomodulatory drug Lenalidomide (1μM for 1hr). Treatment with this agent enhanced the migratory potential of CLL T cells to a speed comparable to untreated and treated healthy T cells. The finding that lenalidomide can restore rapid migration in patient T cells provides evidence that this agent may increase immune surveillance in CLL patients.

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GSK3β Interacts With CRMP2 and Notch1 and Controls T-Cell Motility

Frontiers in Immunology ◽

10.3389/fimmu.2021.680071 ◽

2021 ◽

Vol 12 ◽

Author(s):

Mobashar Hussain Urf Turabe Fazil ◽

Praseetha Prasannan ◽

Brandon Han Siang Wong ◽

Amuthavalli Kottaiswamy ◽

Nur Syazwani Binte Mohamed Salim ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cell Motility ◽

Nuclear Translocation ◽

Glycogen Synthase ◽

Immune Surveillance ◽

Lymphatic Vessels ◽

Adaptive Immune Response ◽

Microtubule Organizing Center ◽

Peripheral Tissues

The trafficking of T-cells through peripheral tissues and into afferent lymphatic vessels is essential for immune surveillance and an adaptive immune response. Glycogen synthase kinase 3β (GSK3β) is a serine/threonine kinase and regulates numerous cell/tissue-specific functions, including cell survival, metabolism, and differentiation. Here, we report a crucial involvement of GSK3β in T-cell motility. Inhibition of GSK3β by CHIR-99021 or siRNA-mediated knockdown augmented the migratory behavior of human T-lymphocytes stimulated via an engagement of the T-cell integrin LFA-1 with its ligand ICAM-1. Proteomics and protein network analysis revealed ongoing interactions among GSK3β, the surface receptor Notch1 and the cytoskeletal regulator CRMP2. LFA-1 stimulation in T-cells reduced Notch1-dependent GSK3β activity by inducing phosphorylation at Ser9 and its nuclear translocation accompanied by the cleaved Notch1 intracellular domain and decreased GSK3β-CRMP2 association. LFA-1-induced or pharmacologic inhibition of GSK3β in T-cells diminished CRMP2 phosphorylation at Thr514. Although substantial amounts of CRMP2 were localized to the microtubule-organizing center in resting T-cells, this colocalization of CRMP2 was lost following LFA-1 stimulation. Moreover, the migratory advantage conferred by GSK3β inhibition in T-cells by CHIR-99021 was lost when CRMP2 expression was knocked-down by siRNA-induced gene silencing. We therefore conclude that GSK3β controls T-cell motility through interactions with CRMP2 and Notch1, which has important implications in adaptive immunity, T-cell mediated diseases and LFA-1-targeted therapies.

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CrkL is required for donor T cell migration to GvHD Target organs

10.1101/854042 ◽

2019 ◽

Author(s):

Nathan H. Roy ◽

Mahinbanu Mammadli ◽

Janis K. Burkhardt ◽

Mobin Karimi

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Adaptor Protein ◽

Cell Transplant ◽

Hematopoietic Stem ◽

Peripheral Tissues ◽

T Cell Migration ◽

Related Family

ABSTRACTThe success of cancer therapies based on allogeneic hematopoietic stem cell transplant relies on the ability to separate graft-versus-host disease (GvHD) from graft-versus-tumor (GVT) responses. Controlling donor T cell migration into peripheral tissues is a viable option to limit unwanted tissue damage, but a lack of specific targets limits progress on this front. Here, we show that the adaptor protein CrkL, but not the closely related family members CrkI or CrkII, is a crucial regulator of T cell migration. In vitro, CrkL-deficient T cells fail to polymerize actin in response to the integrin ligand ICAM-1, resulting in defective migration. Using a mouse model of GvHD/GVT, we found that while CrkL-deficient T cells can efficiently eliminate hematopoietic tumors they are unable to migrate into inflamed organs, such as the liver and small intestine, and thus do not cause GvHD. These results suggest a specific role for CrkL in trafficking to peripheral organs but not the lymphatic system. In line with this, we found that although CrkL-deficient T cells could clear hematopoietic tumors, they failed to clear the same tumor growing subcutaneously, highlighting the role of CrkL in controlling T cell migration into peripheral tissues. Our results define a unique role for CrkL in controlling T cell migration, and suggest that CrkL function could be therapeutically targeted to enhance the efficacy of immunotherapies involving allogeneic donor cells.

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Critical Role of Histone Demethylase Jmjd3 in T Cell Migration

Blood ◽

10.1182/blood-2018-99-110048 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 3708-3708

Author(s):

Chuntang Fu ◽

Qingtian Li ◽

Jia Zou ◽

Changsheng Xing ◽

Bingnan Yin ◽

...

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Cd4 T Cells ◽

Target Gene ◽

Molecular Mechanisms ◽

Critical Role ◽

Altered Expression ◽

T Cell Migration ◽

Body Regions

Abstract Jmjd3, a histone H3K27 demethylase, is known to play a critical role in macrophage and T cell differentiation, but its role in T cell migration and T cell memory maintenance remains largely unknown. In this study, we show that Jmjd3 deficiency resulted in multiple alterations in T cell migration. Jmjd3 deletion limits CD4+ T cells egress out of the thymus, leading to thymic T-cell accumulation and peripheral lymphoid organ T-cell reduction. Gene profiling analysis of wild-type and Jmjd3-deficient CD4+ T cells identified altered expression of Jmjd3 target genes that correlated with changes in H3K27 and/or H3K4 trimethylation in promoters and gene body regions. The expression of the Jmjd3 target gene Pdlim4 is also regulated by Klf2, which regulates T-cell migration. Thus, our findings identify a novel Jmjd3 target gene, Pdlim4, in CD4+ T cell migration and provide insight into the molecular mechanisms by which Jmjd3 regulates T-cell migration. Disclosures No relevant conflicts of interest to declare.

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Molecular Mechanism of Nef-Mediated Inhibition of T Cell Migration in Response to SDF-1α.

Blood ◽

10.1182/blood.v104.11.602.602 ◽

2004 ◽

Vol 104 (11) ◽

pp. 602-602

Author(s):

In-Woo Park ◽

Yumi Mitsuya ◽

Jerome E. Groopman

Keyword(s):

T Cells ◽

Cell Migration ◽

T Cell ◽

Molecular Mechanisms ◽

Transendothelial Migration ◽

Therapeutic Interventions ◽

Sequence Motif ◽

Jurkat T Cells ◽

T Cell Migration ◽

Hiv 1

Abstract Lymphocyte trafficking comprises an intricate multistep process, in which lymphocyte surface integrins, endothelial adhesion molecules, and chemokines produced in the local microenvironment play key roles. We previously reported that HIV-1 nef inhibits T cell chemotaxis in response to the physiological ligand SDF-1αwithout altering CXCR4 expression. Here, we explore the mechanism of this effect. HIV-1 nef significantly reduced the expression of LFA-1, inhibited the adhesion of nef-expressing Jurkat T cells to the HUVEC endothelium, and reduced their transendothelial migration. To gain further insight into how nef mediated these effects, substitution or deletion mutations were introduced into the nef gene, and transwell and transendothelial migration assays were performed. Deletion of the N-terminus proximal basic-rich domain did not change the nef-mediated inhibitory effects on T cell migration, whereas removal of the myristoylation site or the proline-rich amino acid sequence motif of Nef abolished these effects. Similarly, HIV-1 nef blocked the transendothelial migration of Jurkat T cells across the HUVEC monolayer. However, the observed inhibition of transendothelial migration conferred by the wild type nef expression was significantly but not completely reduced by introduction of mutations in the myristoylation site and proline-rich motif, indicating the involvement of other regions in the nef gene. Taken together, these results demonstrate that HIV-1 nef can impair the transendothelial migration of T cells in response to SDF-1αby disturbing different steps in the process, and that the myristoylation site and/or proline-rich motif are key to this effect. These data help to establish the molecular mechanisms of nef-mediated impairment of the T cell migratory response, an essential component of host defense, and provide the basis for targeted therapeutic interventions in patients with AIDS.

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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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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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