IL-28A (IFN-λ2) modulates lung DC function to promote Th1 immune skewing and suppress allergic airway disease.

IL-28 (IFN-λ) cytokines exhibit potent antiviral and antitumor function but their full spectrum of activities remains largely unknown. Recently, IL-28 cytokine family members were found to be profoundly down-regulated in allergic asthma. We now reveal a novel role of IL-28 cytokines in inducing type 1 immunity and protection from allergic airway disease. Treatment of wild-type mice with recombinant or adenovirally expressed IL-28A ameliorated allergic airway disease, suppressed Th2 and Th17 responses and induced IFN-γ. Moreover, abrogation of endogenous IL-28 cytokine function in IL-28Rα(-/-) mice exacerbated allergic airway inflammation by augmenting Th2 and Th17 responses, and IgE levels. Central to IL-28A immunoregulatory activity was its capacity to modulate lung CD11c(+) dendritic cell (DC) function to down-regulate OX40L, up-regulate IL-12p70 and promote Th1 differentiation. Consistently, IL-28A-mediated protection was absent in IFN-γ(-/-) mice or after IL-12 neutralization and could be adoptively transferred by IL-28A-treated CD11c(+) cells. These data demonstrate a critical role of IL-28 cytokines in controlling T cell responses in vivo through the modulation of lung CD11c(+) DC function in experimental allergic asthma. →See accompanying Closeup by Michael R Edwards and Sebastian L Johnston http://dx.doi.org/10.1002/emmm.201100143.

TCR signaling requirements for activating T cells and for generating memory.

Over the last two decades the molecular and cellular mechanisms underlying T cell activation, expansion, differentiation, and memory formation have been intensively investigated. These studies revealed that the generation of memory T cells is critically impacted by a number of factors, including the magnitude of the inflammatory response and cytokine production, the type of dendritic cell [DC] that presents the pathogen derived antigen, their maturation status, and the concomitant provision of costimulation. Nevertheless, the primary stimulus leading to T cell activation is generated through the T cell receptor [TCR] following its engagement with a peptide MHC ligand [pMHC]. The purpose of this review is to highlight classical and recent findings on how antigen recognition, the degree of TCR stimulation, and intracellular signal transduction pathways impact the formation of effector and memory T cells.

Bacterial-induced protection against allergy through a novel multi-component immunoregulatory mechanism

Airborne microbial products have been reported to promote immune responses that suppress asthma, yet how these beneficial effects take place remains controversial and poorly understood. We have found that pulmonary exposure with the bacterium Escherichia coli leads to a suppression of allergic airway inflammation, characterized by reduced airway-hyperresponsiveness, eosinophilia and cytokine production by T cells in the lung. This immune modulation was neither mediated by the induction of a Th1 response nor regulatory T cells; was dependent on TLR-4 but did not involve TLR-desensitization. Dendritic cell migration to the draining lymph nodes and subsequent activation of T cells was unaffected by prior exposure to E.coli indicating that the immunomodulation was limited to the lung environment. In non-treated control mice ovalbumin was primarily presented by airway CD11b+ CD11c+ DCs expressing high levels of MHC class II molecules whilst the DCs in E.coli-treated mice displayed a less activated phenotype and had impaired antigen presentation capacity. Consequently, in situ Th2 cytokine production by ovalbuminspecific effector T cells recruited to the airways was significantly reduced. The suppression of airways hyper responsiveness was mediated through the recruitment of IL-17-producing gd-T cells; however, the suppression of dendritic cells and T cells was mediated through a distinct mechanism that could not be overcome by the local administration of activated dendritic cells, or by the in vivo administration of TNF-alpha. Taken together, these data reveal a novel multi-component immunoregulatory pathway that acts to protect the airways from allergic inflammation.

NLRP3 inflammasome is required in murine asthma in the absence of aluminum adjuvant.

Background: Inflammasome activation with the production of IL-1 beta received substantial attention recently in inflammatory diseases. However, the role of inflammasome in the pathogenesis of asthma is not clear. Using an adjuvant-free model of allergic lung inflammation induced by ovalbumin (OVA), we investigated the role of NLRP3 inflammasome and related it to IL-1R1 signaling pathway.Methods: Allergic lung inflammation induced by OVA was evaluated in vivo in mice deficient in NLRP3 inflammasome, IL-1R1, IL-1 beta or IL-1 alpha. Eosinophil recruitment, Th2 cytokine, and chemokine levels were determined in bronchoalveolar lavage fluid, lung homogenates, and mediastinal lymph node cells ex vivo.Results: Allergic airway inflammation depends on NLRP3 inflammasome activation. Dendritic cell recruitment into lymph nodes, Th2 lymphocyte activation in the lung and secretion of Th2 cytokines and chemokines are reduced in the absence of NLRP3. Absence of NLRP3 and IL-1 beta is associated with reduced expression of other proinflammatory cytokines such as IL-5, IL-13, IL-33, and thymic stromal lymphopoietin. Furthermore, the critical role of IL-1R1 signaling in allergic inflammation is confirmed in IL-1R1-, IL-1 beta-, and IL-1 alpha-deficient mice.Conclusion: NLRP3 inflammasome activation leading to IL-1 production is critical for the induction of a Th2 inflammatory allergic response.

Gut microbiota metabolism of dietary fiber influences allergic airway disease and hematopoiesis.

Metabolites from intestinal microbiota are key determinants of host-microbe mutualism and, consequently, the health or disease of the intestinal tract. However, whether such host-microbe crosstalk influences inflammation in peripheral tissues, such as the lung, is poorly understood. We found that dietary fermentable fiber content changed the composition of the gut and lung microbiota, in particular by altering the ratio of Firmicutes to Bacteroidetes. The gut microbiota metabolized the fiber, consequently increasing the concentration of circulating short-chain fatty acids (SCFAs). Mice fed a high-fiber diet had increased circulating levels of SCFAs and were protected against allergic inflammation in the lung, whereas a low-fiber diet decreased levels of SCFAs and increased allergic airway disease. Treatment of mice with the SCFA propionate led to alterations in bone marrow hematopoiesis that were characterized by enhanced generation of macrophage and dendritic cell (DC) precursors and subsequent seeding of the lungs by DCs with high phagocytic capacity but an impaired ability to promote T helper type 2 (TH2) cell effector function. The effects of propionate on allergic inflammation were dependent on G protein-coupled receptor 41 (GPR41, also called free fatty acid receptor 3 or FFAR3), but not GPR43 (also called free fatty acid receptor 2 or FFAR2). Our results show that dietary fermentable fiber and SCFAs can shape the immunological environment in the lung and influence the severity of allergic inflammation.

Type I IFNs at the interface between cutaneous immunity and epidermal remodeling.

Type I IFNs are key cytokines in antiviral host defense. Preferentially expressed by plasmacytoid dendritic cells, type I IFNs are induced by viral infection and in common skin wounds. In this issue, Tohyama et al. identify a new link between type I IFNs and epidermal remodeling, by showing that type I IFNs specifically upregulate IL-22R expression on keratinocytes and, thereby, IL-22-mediated Stat3 phosphorylation in keratinocytes. The findings suggest that type I IFNs play dual roles in human skin: first, they induce immune activation with the induction of IL-22-producing T cells; second, they provide the interface between immune activation and epidermal remodeling by increasing keratinocyte responsiveness to IL-22.

Functional analysis of monocyte MHC class II compartments.

Circulating monocytes, as dendritic cell and macrophage precursors, exhibit several functions usually associated with antigen-presenting cells, such as phagocytosis and presence of endosomal/lysosomal degradative compartments particularly enriched in Lamp-1, MHC class II molecules, and other proteins related to antigen processing and MHC class II loading [MHC class II compartments (MIICs)]. Ultrastructural analysis of these organelles indicates that, differently from the multivesicular bodies present in dendritic cells, in monocytes the MIICs are characterized by a single perimetral membrane surrounding an electron-dense core. Analysis of their content reveals enrichment in myeloperoxidase, an enzyme classically associated with azurophilic granules in granulocytes and mast cell secretory lysosomes. Elevation in intracellular free calcium levels in monocytes induced secretion of beta-hexosaminidase, cathepsins, and myeloperoxidase in the extracellular milieu; surface up-regulation of MHC class II molecules; and appearance of lysosomal resident proteins. The Ca(2+)-regulated surface transport mechanism of MHC class II molecules observed in monocytes is different from the tubulovesicular organization of the multivesicular bodies previously reported in dendritic cells and macrophages. Hence, in monocytes, MHC class II-enriched organelles combine degradative functions typical of lysosomes and regulated secretion typical of secretory lysosomes. More important, Ca(2+)-mediated up-regulation of surface MHC class II molecules is accompanied by extracellular release of lysosomal resident enzymes.

Cytosolic sensing of extracellular self-DNA transported into monocytes by the antimicrobial peptide LL37.

The intracellular location of nucleic acid sensors prevents recognition of extracellular self-DNA released by dying cells. However, on forming a complex with the endogenous antimicrobial peptide LL37, extracellular DNA is transported into endosomal compartments of plasmacytoid dendritic cells, leading to activation of Toll-like receptor-9 and induction of type I IFNs. Whether LL37 also transports self-DNA into nonplasmacytoid dendritic cells, leading to type I IFN production via other intracellular DNA receptors is unknown. Here we found that LL37 very efficiently transports self-DNA into monocytes, leading the production of type I IFNs in a Toll-like receptor-independent manner. This type I IFN induction was mediated by double-stranded B form DNA, regardless of its sequence, CpG content, or methylation status, and required signaling through the adaptor protein STING and TBK1 kinase, indicating the involvement of cytosolic DNA sensors. Thus, our study identifies a novel link between the antimicrobial peptides and type I IFN responses involving DNA-dependent activation of cytosolic sensors in monocytes.

Classification of current anticancer immunotherapies.

During the past decades, anticancer immunotherapy has evolved from a promising therapeutic option to a robust clinical reality. Many immunotherapeutic regimens are now approved by the US Food and Drug Administration and the European Medicines Agency for use in cancer patients, and many others are being investigated as standalone therapeutic interventions or combined with conventional treatments in clinical studies. Immunotherapies may be subdivided into "passive" and "active" based on their ability to engage the host immune system against cancer. Since the anticancer activity of most passive immunotherapeutics (including tumor-targeting monoclonal antibodies) also relies on the host immune system, this classification does not properly reflect the complexity of the drug-host-tumor interaction. Alternatively, anticancer immunotherapeutics can be classified according to their antigen specificity. While some immunotherapies specifically target one (or a few) defined tumor-associated antigen(s), others operate in a relatively non-specific manner and boost natural or therapy-elicited anticancer immune responses of unknown and often broad specificity. Here, we propose a critical, integrated classification of anticancer immunotherapies and discuss the clinical relevance of these approaches.

Role of neutrophils in "Leishmania" infection

Résumé : Dans le modèle murin d'infection avec le parasite protozoaire Leishmania major (L. major), la souche de souris C57BL/6 est résistante a |'infection et développe une réponse protectrice Thelper (Th) 1. Inversement, les souris de la souche BALB/c développent une réponse Th2 et sont sensibles a cette infection. A la suite d'une infection avec ce parasite, les neutrophiles sont les premières cellules présentes au site d'infection et sont recrutées de manière égale dans les souches résistantes et sensibles à L. major, Néanmoins, trois jours après l'infection, la majorité des neutrophiles disparaissent du site d'infection chez les souris C57BL/6, tandis que ils restent jusqu'a dix jours chez les souris BALB/c. Un rôle crucial des neutrophiles a été démontré durant l'infection avec L. major. En effet, la déplétion de ces cellules avant |'infection dans les souris BALB/c, conduit a une réduction du développement des lésions, associée à une baisse de la charge parasitaire et a une modification de la réponse immunitaire vers une réponse Th1 dans des souris normalement sensibles a |'infection, suggérant un rôle immunorégulateur de ces neutrophiles durant les premiers jours de l'infection. Dans la première partie de cette thèse, nous avons étudié le rôle des neutrophiles suite à l'infection avec L. major. Nous avons démontré que le parasite induisait des phénotypes de neutrophiles distincts chez les souris résistantes ou sensibles à L. major. Suite à l'exposition au parasite, les neutrophiles de souris C57BL/6 ont montré une expression élevée des récepteurs Toll-like 2, 7 et 9 ainsi que la sécrétion d'lL-12p7O et d'lL-10, alors que ceux de souris BALB/c sécrétaient de l'IL-12p40 et du TGFB. Nous avons ensuite démontré qu'en réponse à L. major, au contraire des neutrophiles de BALB/c, les neutrophiles de souris résistantes C57BL/6, libéraient la chimiokine CCL3 attirant les cellules dendritiques. Le rôle crucial de cette chimiokine dans la migration de la première de vague de cellules dendritiques au site d'infection ainsi que son rôle dans le développement de la réponse immunitaire subséquente a été établi. Ces résultats démontrent que les neutrophiles, suite a |'infection avec le parasite L. major, créent un microenvironnement capable de déterminer le développement d'une réponse immunitaire spécifique a un antigène. Dans un second temps, nous nous sommes intéressés au rôle des neutrophiles suite a l'infection avec d'autres espèces de Leishmania: L, doriovani et L. mexicaria, agents responsables de leishmaniose viscérale et cutanée chronique respectivement. Un rôle crucial des neutrophiles a été démontré dans la réponse protectrice suite a l'infection avec L. donovani, l'absence de ces cellules amenant à une susceptibilité au parasite accrue, associée avec une induction préférentielle d'une réponse Th2. Inversement, la déplétion des neutrophiles lors de l'infection avec L. mexicaria aboutit a une résistance accrue, comme constaté par la baisse dela charge parasitaire, la hausse de la réponse Th1 ainsi la baisse de la réponse Th2 dans les souris déplétées en neutrophiles. Néanmoins, malgré le rôle délétère des neutrophiles sur le développement d'une réponse protectrice suite à |'infection avec L. mexicana, ces cellules sont nécessaires pour une résolution correcte dela réponse inflammatoire. En résumé, cette étude révèle un rôle majeur des neutrophiles lors de |'infection avec plusieurs especes de Leishmania. Résumé pour un large public : Les neutrophiles font partie de la famille des globules blancs. A la suite d'une infection, ces cellules sont les premières a être recrutées au site d'infection et sont impliquées dans |'élimination des pathogènes. Dans cette thèse, nous nous somme donc intéressés au rôle que pouvaient jouer ces neutrophiles durant l'infection avec le parasite protozoaire Leishmania major (L. major). Dans le modèle murin d'infection avec L. major, la majorité des souches de souris utilisées dans la recherche, dont les souris de la souche C57BL/6, développent de petites lésions qui guérissent spontanément après quelques semaines (souris résistantes). ll existe néanmoins, quelques souches de souris, dont la souche de souris BALB/c, qui développent des lésions qui ne guérissent pas (souris sensibles). Il a été observé que lors de l'lnfection avec ce parasites les neutrophiles étaient les premières cellules recrutées au site de l'lnfection dans toutes les souches de souris, toutefois trois jours après le début dela réaction immunitaire, la majorité des neutrophiles disparaissent chez les souris C57BL/6, tandis qu'ils restent jusqu'à dix jours chez les souris BALB/c. De plus, un rôle crucial des neutrophiles a été démontré durant l'infection avec L. major. En effet, l'absence de neutrophiles durant les trois premiers jours de l'infection chez les souris sensibles à |'infection, rend ces souris résistantes. Ces résultats suggèrent donc un rôle régulateur de la réponse immunitaire des neutrophiles durant les premiers jours de l'infection. Dans la première partie de cette thèse, nous avons étudié le rôle des neutrophiles suite à l'infection avec L. major. Nous avons donc analysé la sécrétion des cytokines, molécules essentielles qui déterminent la réponse immunitaire, par les neutrophiles. Nous avons démontré que le parasite induisait une sécrétion de cytokines différente entre les souris résistantes ou sensibles a L. major. Nous avons ensuite démontré que seule la souche de souris résistante sécrétait la chimiokine CCL3, connue pour être impliquée dans le recrutement de différentes cellules au site d'infecti0n, dont les cellules dendritiques. Les cellules dendritiques sont un élément fondamental pour un bon déroulement d'une réponse immunitaire, de par leur rôle décisif de liaison entre une réponse précoce non-spécifique au pathogène et une réponse plus tardive spécifique au pathogène et nécessaire pour |'élimination de dernier. Nous avons démontré que les neutrophiles de souris résistantes sécrétaient CCL3 et recrutaient les cellules dendritiques au site d'infecti0n, jouant de ce fait un rôle essentiel dans le développement de la réponse immunitaire. Ces résultats démontrent que les neutrophiles, suite à l'infection avec le parasite L. major, créent un microenvironnement capable de déterminer le développement d'une réponse immunitaire. Dans un second temps, nous nous sommes intéressés au rôle des neutrophiles suite à l'lnfection avec d'autres espèces de Leishmania, L. donovani et L. mexicana. Nous avons pu montrer un rôle crucial de ces cellules dans la réponse à ces deux parasites. En effet, suite à |'infection avec L. donovani, un rôle protecteur des neutrophiles a été observé, leur absence menant à une susceptibilité accrue aux parasites. Dans le cas de l'infection avec L. mexicana, une réduction de |'infection a été observée en absence de neutrophiles, avec néanmoins une augmentation de la lésion, suggérant un rôle important de ces cellules dans le développement de la réponse immunitaire ainsi que dans le contrôle de la réponse inflammatoire. En résumé, cette étude révèle un rôle majeur des neutrophiles lors de l'lnfection avec plusieurs membres de la famille Leishrnania. Summary : Upon infection with the protozoan parasite Leishmania major (L. major), C57BL/6 mice show a resistant phenotype, developing a protective Thelper (Th) 1 response. ln contrast, BALB/c mice develop a Th2 response and are susceptible to infection. Following inoculation with the parasite, neutrophils are the first cells migrating at the site of infection and are equally recruited in both L. major- resistant and susceptible mouse strains. However, after three days of infection, almost all neutrophils disappear from the site of infection in C57BL/6 mice, while they persist until ten days in BALB/c mice. Neutrophils were shown to play a crucial role during infection with L. major. indeed, depletion of these cells in BALB/c mice prior to infection with the parasite led to a lower Iesion development, associated with a lower parasite burden and a modification in the immune response towards a Th1 response in these otherwise susceptible mice, suggesting an immunomodulatory role for neutrophils during the first days of infection. ln the first part of this thesis, we were interested in better understanding the role of neutrophils in infection with L. major. \/\/e found that this parasite was inducing distinct neutrophil phenotypes in L. major-resistant and susceptible mice. Upon exposition with L. major, C57BL/6 neutrophils were reported to express high level of Toll-like receptors 2, 7, 9 mRNA and secrete IL-12p70 and IL-10, while BALB/c neutrophils secreted homodimers of IL-12p40, and TGFB. We then demonstrated that in response to L. major, neutrophils from L. major-resistant C57BL/6 mice release the CCL3 dendritic cell attracting chemokine, which is critical for the first wave of dendritic cell migration to the site of infection and in the development of the subsequent immune response. Altogether, these results demonstrated that upon infection with L. major, neutrophils create a microenvironment that can determine the development of an antigen-specific immune response. ln the second part of the thesis we were interested in understanding the role of neutrophils upon infection with of other species of Leishmania: L. donovani causing visceral leishmaniasis and L. mexicana, agent of chronic cutaneous leishmaniasis. Upon infection with L. donovani, neutrophils were found to play a crucial role in the early protective response, their absence leading to an increased susceptibility to the parasite, associated with the preferential induction of a Th2 response. ln contrast, depletion of these cells early in infection with L. mexicana was leading to an increased resistance, as observed by a decreased parasite burden, increased Th1 and decreased Th2 response in neutrophil-depleted mice. However, despite the deleterious role of neutrophils on the development of a protective immune response upon L. mexicana infection, these cells were required for the proper resolution of the inflammatory response. Altogether, these results highlight a major immunomodulatory role for neutrophils in infection with several species of Leishmania.

Cooperation of human tumor-reactive CD4+ and CD8+ T cells after redirection of their specificity by a high-affinity p53A2.1-specific TCR.

Efficient immune attack of malignant disease requires the concerted action of both CD8+ CTL and CD4+ Th cells. We used human leukocyte antigen (HLA)-A*0201 (A2.1) transgenic mice, in which the mouse CD8 molecule cannot efficiently interact with the alpha3 domain of A2.1, to generate a high-affinity, CD8-independent T cell receptor (TCR) specific for a commonly expressed, tumor-associated cytotoxic T lymphocyte (CTL) epitope derived from the human p53 tumor suppressor protein. Retroviral expression of this CD8-independent, p53-specific TCR into human T cells imparted the CD8+ T lymphocytes with broad tumor-specific CTL activity and turned CD4+ T cells into potent tumor-reactive, p53A2.1-specific Th cells. Both T cell subsets were cooperative and interacted synergistically with dendritic cell intermediates and tumor targets. The intentional redirection of both CD4+ Th cells and CD8+ CTL by the same high-affinity, CD8-independent, tumor-specific TCR could provide the basis for novel broad-spectrum cancer immunotherapeutics.

Caspase-1 autoproteolysis is differentially required for NLRP1b and NLRP3 inflammasome function.

Inflammasomes are caspase-1-activating multiprotein complexes. The mouse nucleotide-binding domain and leucine rich repeat pyrin containing 1b (NLRP1b) inflammasome was identified as the sensor of Bacillus anthracis lethal toxin (LT) in mouse macrophages from sensitive strains such as BALB/c. Upon exposure to LT, the NLRP1b inflammasome activates caspase-1 to produce mature IL-1β and induce pyroptosis. Both processes are believed to depend on autoproteolysed caspase-1. In contrast to human NLRP1, mouse NLRP1b lacks an N-terminal pyrin domain (PYD), indicating that the assembly of the NLRP1b inflammasome does not require the adaptor apoptosis-associated speck-like protein containing a CARD (ASC). LT-induced NLRP1b inflammasome activation was shown to be impaired upon inhibition of potassium efflux, which is known to play a major role in NLRP3 inflammasome formation and ASC dimerization. We investigated whether NLRP3 and/or ASC were required for caspase-1 activation upon LT stimulation in the BALB/c background. The NLRP1b inflammasome activation was assessed in both macrophages and dendritic cells lacking either ASC or NLRP3. Upon LT treatment, the absence of NLRP3 did not alter the NLRP1b inflammasome activity. Surprisingly, the absence of ASC resulted in IL-1β cleavage and pyroptosis, despite the absence of caspase-1 autoprocessing activity. By reconstituting caspase-1/caspase-11(-/-) cells with a noncleavable or catalytically inactive mutant version of caspase-1, we directly demonstrated that noncleavable caspase-1 is fully active in response to the NLRP1b activator LT, whereas it is nonfunctional in response to the NLRP3 activator nigericin. Taken together, these results establish variable requirements for caspase-1 cleavage depending on the pathogen and the responding NLR.

Liquid-crystalline ordering of antimicrobial peptide-DNA complexes controls TLR9 activation.

Double-stranded DNA (dsDNA) can trigger the production of type I interferon (IFN) in plasmacytoid dendritic cells (pDCs) by binding to endosomal Toll-like receptor-9 (TLR9; refs , , , , ). It is also known that the formation of DNA-antimicrobial peptide complexes can lead to autoimmune diseases via amplification of pDC activation. Here, by combining X-ray scattering, computer simulations, microscopy and measurements of pDC IFN production, we demonstrate that a broad range of antimicrobial peptides and other cationic molecules cause similar effects, and elucidate the criteria for amplification. TLR9 activation depends on both the inter-DNA spacing and the multiplicity of parallel DNA ligands in the self-assembled liquid-crystalline complex. Complexes with a grill-like arrangement of DNA at the optimum spacing can interlock with multiple TLR9 like a zipper, leading to multivalent electrostatic interactions that drastically amplify binding and thereby the immune response. Our results suggest that TLR9 activation and thus TLR9-mediated immune responses can be modulated deterministically.

The Ovarian Cancer Chemokine Landscape Is Conducive to Homing of Vaccine-Primed and CD3/CD28-Costimulated T Cells Prepared for Adoptive Therapy.

PURPOSE: Chemokines are implicated in T-cell trafficking. We mapped the chemokine landscape in advanced stage ovarian cancer and characterized the expression of cognate receptors in autologous dendritic cell (DC)-vaccine primed T cells in the context of cell-based immunotherapy. EXPERIMENTAL DESIGN: The expression of all known human chemokines in patients with primary ovarian cancer was analyzed on two independent microarray datasets and validated on tissue microarray. Peripheral blood T cells from five HLA-A2 patients with recurrent ovarian cancer, who previously received autologous tumor DC vaccine, underwent CD3/CD28 costimulation and expansion ex vivo. Tumor-specific T cells were identified by HER2/neu pentamer staining and were evaluated for the expression and functionality of chemokine receptors important for homing to ovarian cancer. RESULTS: The chemokine landscape of ovarian cancer is heterogeneous with high expression of known lymphocyte-recruiting chemokines (CCL2, CCL4, and CCL5) in tumors with intraepithelial T cells, whereas CXCL10, CXCL12, and CXCL16 are expressed quasi-universally, including in tumors lacking tumor-infiltrating T cells. DC-vaccine primed T cells were found to express the cognate receptors for the above chemokines. Ex vivo CD3/CD28 costimulation and expansion of vaccine-primed Tcells upregulated CXCR3 and CXCR4, and enhanced their migration toward universally expressed chemokines in ovarian cancer. CONCLUSIONS: DC-primed tumor-specific T cells are armed with the appropriate receptors to migrate toward universal ovarian cancer chemokines, and these receptors are further upregulated by ex vivo CD3/CD28 costimulation, which render T cells more fit for migrating toward these chemokines. Clin Cancer Res; 21(12); 2840-50. ©2015 AACR.