Dendritic cells pulsed with Pythium insidiosum (1,3)(1,6)-β-glucan, Heat-inactivated zoospores and immunotherapy prime naïve T cells to Th1 differentiation in vitro

Because of the low frequency of T cells for any particular soluble protein antigen in unprimed animals, the requirements for naive T cell responses in specific antigens have not been clearly delineated and they have been difficult to study in vitro. We have taken advantage of mice transgenic for the V beta 3/V alpha 11 T cell receptor (TCR), which can recognize a peptide of cytochrome c presented by IEk. 85-90% of CD4+ T cells in these mice express the transgenic TCR, and we show that almost all such V beta 3/V alpha 11 receptor-positive cells have a phenotype characteristic of naive T cells, including expression of high levels of CD45RB, high levels of L-selectin (Mel-14), low levels of CD44 (Pgp-1), and secretion of interleukin 2 (IL-2) as the major cytokine. Naive T cells, separated on the basis of CD45RB high expression, gave vigorous responses (proliferation and IL-2 secretion) to peptide antigen presented in vitro by a mixed antigen-presenting cell population. At least 50% of the T cell population appeared to respond, as assessed by blast transformation, entry into G1, and expression of increased levels of CD44 by 24 h. Significant contributions to the response by contaminating memory CD4+ cells were ruled out by demonstrating that the majority of the CD45RB low, L-selectin low, CD44 high cells did not express the V beta 3/V alpha 11 TCR and responded poorly to antigen. We find that proliferation and IL-2 secretion of the naive CD4 cells is minimal when resting B cells present peptide antigen, and that both splenic and bone marrow-derived macrophages are weak stimulators. Naive T cells did respond well to high numbers of activated B cells. However, dendritic cells were the most potent stimulators of proliferation and IL-2 secretion at low cell numbers, and were far superior inducers of IL-2 at higher numbers. These studies establish that naive CD4 T cells can respond vigorously to soluble antigen and indicate that maximal stimulation can be achieved by presentation of antigen on dendritic cells. This model should prove very useful in further investigations of activation requirements and functional characteristics of naive helper T cells.

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ICAM-1 on exosomes from mature dendritic cells is critical for efficient naive T-cell priming

Blood ◽

10.1182/blood-2005-01-0220 ◽

2005 ◽

Vol 106 (1) ◽

pp. 216-223 ◽

Cited By ~ 300

Author(s):

Elodie Segura ◽

Carole Nicco ◽

Bérangère Lombard ◽

Philippe Véron ◽

Graça Raposo ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

T Cell ◽

Mhc Class Ii ◽

T Cell Responses ◽

Class Ii ◽

Naive T Cells ◽

Naïve T Cells ◽

Cell Responses

Exosomes are secreted vesicles formed in late endocytic compartments. Immature dendritic cells (DCs) secrete exosomes, which transfer functional major histocompatibility complex (MHC)–peptide complexes to other DCs. Since immature and mature DCs induce different functional T-cell responses (ie, tolerance versus priming), we asked whether DC maturation also influenced the priming abilities of their exosomes. We show that exosomes secreted by lipopolysaccharide (LPS)–treated mature DCs are 50- to 100-fold more potent to induce antigen-specific T-cell activation in vitro than exosomes from immature DCs. In vitro, exosomes from mature DCs transfer to B lymphocytes the ability to prime naive T cells. In vivo, only mature exosomes trigger effector T-cell responses, leading to fast skin graft rejection. Proteomic and biochemical analyses revealed that mature exosomes are enriched in MHC class II, B7.2, intercellular adhesion molecule 1 (ICAM-1), and bear little milk-fat globule–epidermal growth factor–factor VIII (MFG-E8) as compared with immature exosomes. Functional analysis using DC-derived exosomes from knock-out mice showed that MHC class II and ICAM-1 are required for mature exosomes to prime naive T cells, whereas B7.2 and MFG-E8 are dispensable. Therefore, changes in protein composition and priming abilities of exosomes reflect the maturation signals received by DCs.

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Investigating the Role of Dendritic Cells in Extracorporeal Photopheresis Using an In Vitro Model.

Blood ◽

10.1182/blood.v108.11.5162.5162 ◽

2006 ◽

Vol 108 (11) ◽

pp. 5162-5162

Author(s):

Udo Holtick ◽

Scott R. Marshall ◽

Muzlifah Haniffa ◽

Catharien M. Hilkens ◽

Xiao N. Wang ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Chronic Gvhd ◽

Extracorporeal Photopheresis ◽

Hematopoietic Stem ◽

Naive T Cells ◽

Naïve T Cells ◽

Cytokine Analysis ◽

Secondary Stimulation

Abstract Graft versus Host disease (GvHD) is the major limitation to successful allogeneic hematopoietic stem cell transplantation and contributes significantly to transplant related mortality and morbidity. Steroid refractory or steroid depending GvHD in particular is linked to poor survival or life quality. Conventional immunosuppression has very limited success in these conditions and increases susceptability for infection and relapse. Extracorporeal Photopheresis (ECP) is a promising therapy for acute and chronic GvHD not responding to conventional immunosuppressive therapy. ECP treatment seems not to result in a pan-immunosuppression but has quite selective effects on the pathogenic process in GvHD. The mechanisms of action of ECP in GvHD known so far include lymphocyte apoptosis, cytokine modulation and increased regulatory T cell numbers. It has been suggested that monocytes and dendritic cells (DC) were more resistant to ECP induced apoptosis, so that they might either be directly modulated by ECP or modulated by encounter with apoptotic cells and this way aquiring immunomodulating properties. Here we tested the first hypothesis analysing direct effects of ECP on monocyte derived dendritic cells in vitro. Direct in vitro PUVA treatment leads to activation of monocyte derived immature DCs (upregulation of CD83, CD86, HLA-DR, reduced endocytosis capacity, increased migratory capacity), whereas mature DCs are not affected. However, immature and mature DCs undergo apoptosis later on within 24h-72h. Prestimualtion with either antigen (KLH), IL-12 or co-culture with CD40L expressing cells could not prevent apoptosis. Further DC stimulation through CD40L is abrogated 24h after treatment. DC cytokine analysis (IL-12, TNFα) is pending. After in vitro PUVA primary dermal DCs and Langerhans Cells migrated from skin biopsies undergo apoptosis similarly to monocyte derived DCs. Going along with the early activation, the stimulatory capacity of in vitro PUVA treated immature DCs is enhanced in an MLR on naive T cells. IL-10 levels are decreased as compared to untreated or UVA treated cells. Secondary stimulation through either CD3/CD28 beads or mature DCs leads to reduced proliferation of naive T cells that were primarily stimulated with in vitro PUVA treated as compared to untreated immature DCs. Cytokine analysis after secondary stimulation is pending. We conclude that in our model in vitro PUVA modulates the character especially of immature dendritic cells. However, immunostimulating and immunosuppressive characteristics could be described depending on the assay. Cytokine results after second stimulation (Th1/Th2) are pending. Apoptosis is a major finding and occurs both in monocytes and DCs after 24–72h. Our findings indicate that there might be direct ECP effects on monocytes and DCs going beyond the process of merely depleting antigen presenting cells. Further relevance is currently being investigated.

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CD8α+ and CD8α− Subclasses of Dendritic Cells Direct the Development of Distinct T Helper Cells In Vivo

Journal of Experimental Medicine ◽

10.1084/jem.189.3.587 ◽

1999 ◽

Vol 189 (3) ◽

pp. 587-592 ◽

Cited By ~ 757

Author(s):

Roberto Maldonado-López ◽

Thibaut De Smedt ◽

Patrick Michel ◽

Jacques Godfroid ◽

Bernard Pajak ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Critical Role ◽

Interleukin 12 ◽

T Helper ◽

Naive T Cells ◽

Naïve T Cells ◽

Physiologic Function

Cells of the dendritic family display some unique properties that confer to them the capacity to sensitize naive T cells in vitro and in vivo. In the mouse, two subclasses of dendritic cells (DCs) have been described that differ by their CD8α expression and their localization in lymphoid organs. The physiologic function of both cell populations remains obscure. Studies conducted in vitro have suggested that CD8α+ DCs could play a role in the regulation of immune responses, whereas conventional CD8α− DCs would be more stimulatory. We report here that both subclasses of DCs efficiently prime antigen-specific T cells in vivo, and direct the development of distinct T helper (Th) populations. Antigen-pulsed CD8α+ and CD8α− DCs are separated after overnight culture in recombinant granulocyte/macrophage colony-stimulating factor and injected into the footpads of syngeneic mice. Administration of CD8α− DCs induces a Th2-type response, whereas injection of CD8α+ DCs leads to Th1 differentiation. We further show that interleukin 12 plays a critical role in Th1 development by CD8α+ DCs. These findings suggest that the nature of the DC that presents the antigen to naive T cells may dictate the class selection of the adaptative immune response.

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Granulocyte-colony stimulating factor drives the in vitro differentiation of human dendritic cells that induce anergy in naïve T cells

European Journal of Immunology ◽

10.1002/eji.201040659 ◽

2010 ◽

Vol 40 (11) ◽

pp. 3097-3106 ◽

Cited By ~ 35

Author(s):

Maura Rossetti ◽

Silvia Gregori ◽

Maria Grazia Roncarolo

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Granulocyte Colony Stimulating Factor ◽

Colony Stimulating Factor ◽

In Vitro Differentiation ◽

Naive T Cells ◽

Naïve T Cells ◽

Human Dendritic Cells ◽

Stimulating Factor

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Efficient migration of dendritic cells toward lymph node chemokines and induction of TH1 responses require maturation stimulus and apoptotic cell interaction

Blood ◽

10.1182/blood-2004-10-3991 ◽

2005 ◽

Vol 106 (5) ◽

pp. 1734-1741 ◽

Cited By ~ 26

Author(s):

Nicolas Bertho ◽

Henri Adamski ◽

Louis Toujas ◽

Martine Debove ◽

Jean Davoust ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Lymph Node ◽

Immune Responses ◽

Tumor Cells ◽

Apoptotic Cell ◽

Apoptotic Cells ◽

Naive T Cells ◽

Naïve T Cells ◽

Tnf Α

Abstract Dendritic cells (DCs) have the unique ability to initiate primary immune responses, and they can be conditioned for vaccinal purposes to present antigens after the engulfment of apoptotic cells. To recruit the rare antigen-specific naive T cells, DCs require a maturation step and subsequent transport toward lymph node (LN). To date, prostaglandin E2 (PGE2) is the best-characterized compound inducing this LN-directed migration in vitro, but PGE2 may skew the immune responses in a TH2 direction. We demonstrate here that on incubation with apoptotic tumor cells and tumor necrosis factor-α (TNF-α) or lipopolysaccharide (LPS), human monocyte-derived DCs become fully mature and acquire high migratory capacities toward LN-directing chemokines. The migration of TNF-α-treated DCs occurs only after cotreatment with apoptotic cells but not with necrotic cells. DC migration requires CD36 expression and incubation with apoptotic cells in the presence of heat-labile serum components. Moreover, on treatment with apoptotic cells and LPS, the migrating DCs are able to recruit naive T cells to generate TH1 immune responses. Our results show that the cotreatment of DCs with apoptotic tumor cells and inflammatory signals is promising for the design of an antitumoral DC-based vaccine. (Blood. 2005;106:1734-1741)

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In vitro responses of human CD45R0brightRA− and CD45R0−RAbright T cell subsets and their relationship to memory and naive T cells

European Journal of Immunology ◽

10.1002/eji.1830270937 ◽

1997 ◽

Vol 27 (9) ◽

pp. 2383-2390 ◽

Cited By ~ 72

Author(s):

Joyce L. Young ◽

Judith M. Ramage ◽

J. S. Hill Gaston ◽

Peter C. L. Beverley

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Subsets ◽

Naive T Cells ◽

Naïve T Cells

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Wogonin Attenuates Ovalbumin Antigen-Induced Neutrophilic Airway Inflammation by Inhibiting Th17 Differentiation

International Journal of Inflammation ◽

10.1155/2014/571508 ◽

2014 ◽

Vol 2014 ◽

pp. 1-8 ◽

Cited By ~ 11

Author(s):

Rie Takagi ◽

Masaaki Kawano ◽

Kazuyuki Nakagome ◽

Kumiko Hashimoto ◽

Takehiro Higashi ◽

...

Keyword(s):

T Cells ◽

Airway Inflammation ◽

Th17 Cells ◽

Allergic Airway Inflammation ◽

Naive T Cells ◽

Naïve T Cells ◽

Th17 Differentiation ◽

Kampo Extracts ◽

Human And Mouse

Allergic airway inflammation is generally considered to be a Th2-type immune response. Recent studies, however, have demonstrated that Th17-type immune responses also play important roles in this process, particularly in the pathogenesis of neutrophilic airway inflammation, a hallmark of severe asthma. We scrutinized several Kampo extracts that reportedly exhibit anti-inflammatory activity by usingin vitrodifferentiation system of human and mouse naïve T cells. We found that hange-shashin-to (HST) and oren-gedoku-to (OGT) possess inhibitory activity for Th17 responsesin vitro. Indeed, wogonin and berberine, major components common to HST and OGT, exhibit Th17-inhibitory activities in both murine and human systemsin vitro. We therefore evaluated whether wogonin suppresses OVA-induced neutrophilic airway inflammation in OVA TCR-transgenic DO11.10 mice. Consequently, oral administration of wogonin significantly improved OVA-induced neutrophilic airway inflammation. Wogonin suppressed the differentiation of naïve T cells to Th17 cells, while showing no effects on activated Th17 cells.

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The influence of chorionic gonadotropin on phenotype conversion and hTERT gene expression by T-lymphocytes of different degrees of differentiation

Biomeditsinskaya Khimiya ◽

10.18097/pbmc20176306539 ◽

2017 ◽

Vol 63 (6) ◽

pp. 539-545 ◽

Cited By ~ 1

Author(s):

M.B. Rayev ◽

S.A. Zamorina ◽

L.S. Litvinova ◽

K.A. Yurova ◽

O.G. Khaziakhmatova ◽

...

Keyword(s):

T Cells ◽

T Lymphocytes ◽

Chorionic Gonadotropin ◽

Memory T Cells ◽

Immune Memory ◽

Proliferation Marker ◽

Naive T Cells ◽

Naïve T Cells ◽

Htert Gene

The effects of chorionic gonadotropin (hCG) on the expression of the hTERT gene in combination with the conversion of the phenotype of naive T-cells and T-cells of immune memory in vitro were studied. hCG inhibited expression of hTERT mRNA in naive T-cells (CD45RA+) and immune memory T cells (CD45RO+), causing a decrease in the replicative potential of the cells. The presence of hCG in the culture led to the conversion of the phenotype of T-lymphocytes. hCG reduced the number of proliferating T-cells of immune memory, estimated by phenotypic signs by differential gating. hCG (10 IU/ml and 100 IU/ml) inhibited expression of CD25 by the studied populations, but did not modulate expression of the CD71 proliferation marker. Thus, hCG inhibited the functional activity of naive T-cells and T-cells of immune memory, which, in the context of pregnancy, can contribute to the formation of immune tolerance to the semi-allogenic fetus.

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