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1.
Genes Immun ; 14(1): 35-41, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23151488

ABSTRACT

We have previously described SEG/Pas as the first mouse inbred strain able to survive subcutaneous injection of virulent Yersinia pestis, the agent of plague, and we identified Yprl1, Yprl2 and Yprl3 as three quantitative trait loci (QTLs) controlling this exceptional phenotype in females from a backcross between SEG/Pas and C57BL/6 strains. We have now developed congenic strains to further characterize the extent and effect of these genomic regions. In this study, we confirm the importance of two of these regions, both in males and females, while the third one may well be a spurious association. We show that no genomic region alone is able to increase the survival of C57BL/6 mice, but that C57BL/6 mice carrying both Yprl2 and Yprl3 exhibit intermediate resistance. Each of these two QTLs contains at least two subregions, which are required to increase survival. Finally, through the analysis of congenic strains in an F1 background, we establish the mode of inheritance of the SEG-derived resistance alleles. Altogether, this study has clarified and enhanced our understanding of the genetic architecture of resistance to plague in SEG/Pas mice.


Subject(s)
Disease Resistance/genetics , Plague/genetics , Quantitative Trait Loci , Alleles , Animals , Disease Progression , Female , Male , Mice , Mice, Congenic , Mice, Inbred C57BL , Plague/immunology , Plague/microbiology , Yersinia pestis
2.
Eur J Cancer ; 37(13): 1709-18, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11527700

ABSTRACT

The product of the Lymphocyte Activation Gene-3 (LAG-3, CD223) is a high affinity MHC class II ligand expressed by activated CD4(+) and CD8(+) T cells, which can associate with the T cell receptor (TCR) and downregulate TCR signalling in vitro. We have also reported that a soluble mLAG-3Ig fusion protein works as a vaccine adjuvant in vivo in mice, enhancing Th1 and CD8 T cell responses. Here, we report that LAG-3 expression was found, using fluorescent activated cell sorting (FACS) analysis, on 11-48% of human tumour-infiltrating lymphocytes (TILs) isolated from eight freshly dissociated renal cell carcinomas (RCCs), and was restricted mostly to CD8(+) cells. Immunohistochemical analysis confirmed LAG-3 expression by TILs in 9/11 RCCs, as well as in tumours of different origins, such as melanomas (3/5) and lymphomas (7/7). Since not only antigen presenting cells (APCs), but also TILs themselves strongly express major histocompatibility complex (MHC) class II, we firstly investigated whether LAG-3/MHC class II T-T cell contacts might influence tumour cell recognition. However, cytotoxicity inhibition was not observed in two RCC-specific CD8(+) T cell clones in the presence of the LAG-3-specific MAb, and there was also no observed difference in the recognition of LAG-3-transfected or wild-type RCC by these cytotoxic T lymphocytes (CTLs). In contrast, MHC class II engagement by LAG-3Ig was found to enhance the capacity of immature dendritic cells to stimulate naive T cell proliferation and IL-12-dependent IFN-gamma production by T cells in vitro. These results therefore provide support for a role for TIL-expressed LAG-3 in the engagement of class II molecules on APCs, thereby contributing to APC activation and Th1/Tc1 commitment, without downregulating cytotoxicity.


Subject(s)
Antigens, CD , Genes, MHC Class II/immunology , Membrane Proteins/metabolism , Neoplasms/immunology , T-Lymphocytes/immunology , CD3 Complex/metabolism , CD4-Positive T-Lymphocytes/immunology , Carcinoma, Renal Cell/immunology , Cell Communication/immunology , Dendritic Cells/immunology , Flow Cytometry , Humans , Immunohistochemistry , Kidney Neoplasms/immunology , Lymphocyte Activation/immunology , T-Lymphocytes, Cytotoxic/immunology , Tumor Cells, Cultured , Lymphocyte Activation Gene 3 Protein
3.
J Immunol ; 167(5): 2547-54, 2001 Sep 01.
Article in English | MEDLINE | ID: mdl-11509594

ABSTRACT

Proinflammatory molecules, including IFN-gamma and IL-12, play a crucial role in the elimination of causative agents. To allow healing, potent anti-inflammatory processes are required to down-regulate the inflammatory response. In this study, we first show that CD47/integrin-associated protein, a ubiquitous multispan transmembrane protein highly expressed on T cells, interacts with signal-regulator protein (SIRP)-alpha, an immunoreceptor tyrosine-based inhibition motif-containing molecule selectively expressed on myelomonocytic cells, and next demonstrate that this pair of molecules negatively regulates human T and dendritic cell (DC) function. CD47 ligation by CD47 mAb or L-SIRP-alpha transfectants inhibits IL-12R expression and down-regulates IL-12 responsiveness of activated CD4(+) and CD8(+) adult T cells without affecting their response to IL-2. Human CD47-Fc fusion protein binds SIRP-alpha expressed on immature DC and mature DC. SIRP-alpha engagement by CD47-Fc prevents the phenotypic and functional maturation of immature DC and still inhibits cytokine production by mature DC. Finally, in allogeneic MLR between mDC and naive T cells, CD47-Fc decreases IFN-gamma production after priming and impairs the development of a Th1 response. Therefore, CD47 on T cells and its cognate receptor SIRP-alpha on DC define a novel regulatory pathway that may be involved in the maintenance of homeostasis by preventing the escalation of the inflammatory immune response.


Subject(s)
Antigens, CD/metabolism , Antigens, Differentiation , Carrier Proteins/metabolism , Dendritic Cells/immunology , Interleukin-12/biosynthesis , Membrane Glycoproteins/immunology , Neural Cell Adhesion Molecule L1 , Neural Cell Adhesion Molecules/immunology , Receptors, Immunologic/metabolism , T-Lymphocytes/immunology , Antibodies, Monoclonal/pharmacology , CD47 Antigen , Cell Differentiation , Dendritic Cells/cytology , Down-Regulation , Humans , In Vitro Techniques , Interferon-gamma/biosynthesis , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Neural Cell Adhesion Molecules/genetics , Neural Cell Adhesion Molecules/metabolism , Receptors, Immunologic/genetics , Signal Transduction , Transfection
4.
J Exp Med ; 192(3): 405-12, 2000 Aug 07.
Article in English | MEDLINE | ID: mdl-10934228

ABSTRACT

The subset of dendritic cells (DCs) and the nature of the signal inducing DC maturation determine the capacity of DCs to generate polarized immune responses. In this study, we show that the ability of human monocyte-derived DCs (myeloid DC(1)) to promote T helper type 1 (Th1) or Th2 differentiation was also found to be critically dependent on stimulator/responder ratio. At a low ratio (1:300), mature DCs that have been differentiated after inflammatory (Staphylococcus aureus Cowan 1 or lipopolysaccharide) or T cell-dependent (CD40 ligand) stimulation induced naive T cells to become Th2 (interleukin [IL]-4(+), IL-5(+), interferon gamma) effectors. Th2 differentiation was dependent on B7-CD28 costimulation and enhanced by OX40-OX40 ligand interactions. However, high DC/T cell ratio (1:4) favored a mixed Th1/Th2 cell development. Thus, the fact that the same DC lineage stimulates polarized Th1 or Th2 responses may be relevant since it allows the antigen-presenting cells to initiate an appropriate response for the signal received at the peripheral sites. Controlling the number and the rate of DC migration to the T cell areas in lymphoid tissues may be important for the therapeutic use of DCs.


Subject(s)
Dendritic Cells/metabolism , Leukopoiesis , Membrane Glycoproteins , Th1 Cells/cytology , Th2 Cells/cytology , Animals , B7-1 Antigen/metabolism , CD28 Antigens/metabolism , Dendritic Cells/cytology , Humans , Interleukin-12/metabolism , Mice , Monocytes/cytology , Monocytes/metabolism , OX40 Ligand , Receptors, Immunologic/metabolism , Receptors, OX40 , Receptors, Tumor Necrosis Factor/metabolism , Th1 Cells/metabolism , Th2 Cells/metabolism , Tumor Necrosis Factor Receptor Superfamily, Member 7/metabolism , Tumor Necrosis Factor-alpha/metabolism , Tumor Necrosis Factors
5.
J Immunol ; 164(4): 2193-9, 2000 Feb 15.
Article in English | MEDLINE | ID: mdl-10657674

ABSTRACT

Upon encounter with bacterial products, immature dendritic cells (iDCs) release proinflammatory cytokines and develop into highly stimulatory mature DCs. In the present study, we show that human monocyte-derived DCs functionally express the CD47 Ag, a thrombospondin receptor. Intact or F(ab')2 of CD47 mAb suppress bacteria-induced production of IL-12, TNF-alpha, GM-CSF, and IL-6 by iDCs. 4N1K, a peptide derived from the CD47-binding site of thrombospondin, also inhibits cytokine release. The inhibition of IL-12 and TNF-alpha is IL-10-independent inasmuch as IL-10 production is down-modulated by CD47 mAb and blocking IL-10 mAb fails to restore cytokine levels. CD47 ligation counteracts the phenotypic and functional maturation of iDCs in that it prevents the up-regulation of costimulatory molecules, the loss of endocytic activity, and the acquisition of an increased capacity to stimulate T cell proliferation and IFN-gamma production. Interestingly, regardless of CD47 mAb treatment during DC maturation, mature DC restimulated by soluble CD40 ligand and IFN-gamma, to mimic DC/T interaction, produce less IL-12 and more IL-18 than iDCs. Finally, CD47 ligation on iDCs does not impair their capacity to phagocytose apoptotic cells. We conclude that following exposure to microorganisms, CD47 ligation may limit the intensity and duration of the inflammatory response by preventing inflammatory cytokine production by iDCs and favoring their maintenance in an immature state.


Subject(s)
Antigens, CD/immunology , Antigens, CD/metabolism , Carrier Proteins/immunology , Carrier Proteins/metabolism , Cytokines/antagonists & inhibitors , Cytokines/biosynthesis , Dendritic Cells/cytology , Dendritic Cells/immunology , Antibodies, Monoclonal/metabolism , Antibodies, Monoclonal/pharmacology , Antigens, CD/biosynthesis , CD47 Antigen , Carrier Proteins/biosynthesis , Cell Differentiation/immunology , Cytokines/metabolism , Dendritic Cells/metabolism , Humans , Immunophenotyping , Interferon-gamma/physiology , Interleukin-10/physiology , Ligands , Phagocytosis/immunology , Thrombospondins/metabolism
6.
Int Arch Allergy Immunol ; 118(2-4): 384-6, 1999.
Article in English | MEDLINE | ID: mdl-10224454

ABSTRACT

Our previous studies have indicated that naive human CD4+ T cells of neonatal or adult origin may be the initial source of IL-4 which is required for their development into Th2 effectors. In addition to minute amounts of IL-4, anti-CD3/B7.1-activated naive cells also release readily detectable levels of IL-13 and IFN-gamma. The production of IL-4 and IL-13 by naive T cells is differentially regulated by TGF-beta and IL-12. Shortly after activation, naive T cells express surface OX40, a TNF-R family member whose ligand (OX40L) is constitutively expressed on a subset of dendritic cells. Engagement of OX40 on activated naive T cells increases their expression of IL-4 and IL-13, suppresses that of IFN-gamma and promotes their development into Th2-like effectors.


Subject(s)
CD4-Positive T-Lymphocytes/immunology , Interleukin-4/biosynthesis , Lymphocyte Activation/immunology , Membrane Glycoproteins , Receptors, Tumor Necrosis Factor/immunology , Adult , CD4-Positive T-Lymphocytes/cytology , Cell Differentiation/immunology , Humans , Interleukin-4/immunology , Ligands , Lymphocyte Activation/drug effects , OX40 Ligand , Tumor Necrosis Factor Receptor Superfamily, Member 7/immunology
7.
J Immunol ; 162(7): 3790-4, 1999 Apr 01.
Article in English | MEDLINE | ID: mdl-10201895

ABSTRACT

It is generally accepted that immunologically naive T cells display a very restricted cytokine production profile consisting mainly of IL-2, which is used as an autocrine growth factor. Here we report that activated naive CD4+ T cells, of neonatal or adult origin, express very high levels of soluble lymphotoxin (LT) alpha (LTalpha3), as determined by ELISA, RNase protection assay, and intracytoplasmic staining. Besides LTalpha3 and IL-2, these cells also produce high levels of TNF-alpha together with significant amounts of IFN-gamma and IL-13. Naive cells also express LTbeta mRNA and the membrane form of LTalpha (LTalphabeta). On average, naive CD4+ T cells secrete four times more LTalpha3 than Th1-like cells, twice more than naive CD8+ T cells, and ten times more than B cells. Thus, naive T cells express a large spectrum of cytokines, mainly of the Th1 type, and the very high levels of LTalpha3/TNF-alpha that they release may play an hitherto unsuspected role in the early stage of T cell-dependent immune responses.


Subject(s)
CD4-Positive T-Lymphocytes/metabolism , Lymphotoxin-alpha/metabolism , Adult , CD4-Positive T-Lymphocytes/immunology , Cell Line , Cells, Cultured , Humans , Infant, Newborn , Interferon-gamma/metabolism , Interleukin-13/metabolism , Interphase/immunology , Lymphotoxin-alpha/biosynthesis , Solubility , T-Lymphocyte Subsets/immunology , T-Lymphocyte Subsets/metabolism , Tumor Necrosis Factor-alpha/metabolism
8.
J Immunol ; 162(5): 2748-53, 1999 Mar 01.
Article in English | MEDLINE | ID: mdl-10072520

ABSTRACT

Lymphocyte activation gene-3 (LAG-3) is an MHC class II ligand structurally and genetically related to CD4. Although its expression is restricted to activated T cells and NK cells, the functions of LAG-3 remain to be elucidated. Here, we report on the expression and function of LAG-3 on proinflammatory bystander T cells that are activated in the absence of TCR engagement. LAG-3 is expressed at high levels on human T cells cocultured with autologous monocytes and IL-2 and synergizes with the low levels of CD40 ligand (CD40L) expressed on these cells to trigger TNF-alpha and IL-12 production by monocytes. Indeed, anti-LAG-3 mAb inhibits both IL-12 and IFN-gamma production in IL-2-stimulated cocultures of T cells and autologous monocytes. Soluble LAG-3Ig fusion protein markedly enhances IL-12 production by monocytes stimulated with infra-optimal concentrations of sCD40L, whereas it directly stimulates monocyte-derived dendritic cells (DC) for the production of TNF-alpha and IL-12, unravelling an enhanced responsiveness to MHC class II engagemenent in DC as compared with activated monocytes. Thus similar to CD40L, LAG-3 may be involved in the proinflammatory activity of cytokine-activated bystander T cells and most importantly it may directly activate DC.


Subject(s)
Antigens, CD , Dendritic Cells/physiology , Interleukin-12/biosynthesis , Lymphocyte Activation , Membrane Proteins/physiology , Monocytes/physiology , T-Lymphocytes/immunology , Tumor Necrosis Factor-alpha/biosynthesis , Adult , Animals , CD40 Ligand , CHO Cells , Cricetinae , Humans , Interleukin-2/pharmacology , Membrane Glycoproteins/physiology , Membrane Proteins/genetics , Lymphocyte Activation Gene 3 Protein
9.
J Immunol ; 161(7): 3408-15, 1998 Oct 01.
Article in English | MEDLINE | ID: mdl-9759858

ABSTRACT

At inflammatory sites, the number of activated bystander T cells exceeds that of Ag-activated T cells. We investigated whether IL-15, a monocyte-derived cytokine that shares several biologic activities with IL-2, may contribute to bystander T cell activation in the absence of IL-2 and triggering Ag. The addition of IL-15 to cocultures of monocytes and T cells stimulates CD4+ but not CD8+ T cells to produce IFN-gamma. IFN-gamma production requires endogenous IL-12, the production of which in turn is dependent upon CD40/CD154 interactions between CD4+ T cells and monocytes. Indeed, non-TCR-activated CD4+ but not CD8+ T cells express significant levels of CD154. IL-15 may enhance IFN-gamma in this system by up-regulating CD40 expression on monocytes and IL-12Rbeta1 expression on CD4+ T cells. Conversely, using neutralizing anti-IL-15 mAb, we show that the ability of IL-12 to augment IFN-gamma secretion is partly mediated by endogenous IL-15. Finally, in the absence of monocytes, a synergistic effect between exogenous IL-12 and IL-15 is necessary to induce IFN-gamma production by purified CD4+ T cells, while IL-15 alone induces T cell proliferation. It is proposed that this codependence between IL-12 and IL-15 for the activation of inflammatory T cells may be involved in chronic inflammatory disorders that are dominated by a Th1 response. In such a response, a self-perpetuating cycle of inflammation is set forth, because IL-15-stimulated CD4+ T cells may activate monocytes to release IL-12 that synergizes with IL-15 to induce IL-12 response and IFN-gamma production.


Subject(s)
Adjuvants, Immunologic/pharmacology , CD4-Positive T-Lymphocytes/metabolism , Cell Communication/immunology , Interferon-gamma/metabolism , Interleukin-12/biosynthesis , Interleukin-15/pharmacology , Monocytes/metabolism , Receptors, Antigen, T-Cell/metabolism , Antigens/immunology , CD4-Positive T-Lymphocytes/immunology , CD40 Antigens/physiology , CD40 Ligand , Cells, Cultured , Coculture Techniques , Humans , Interleukin-12/metabolism , Interleukin-12/physiology , Ligands , Membrane Glycoproteins/physiology , Receptors, Antigen, T-Cell/immunology
10.
Vaccine ; 16(14-15): 1415-9, 1998.
Article in English | MEDLINE | ID: mdl-9711781

ABSTRACT

The increased susceptibility of neonates to infections has been ascribed to the immaturity of their immune system. More particularly, T cell-dependent responses were shown to be biased towards a Th2 phenotype. Our studies on the in vitro maturation of umbilical cord blood T cells suggest that the Th2 bias of neonatal response cannot be simply ascribed to intrinsic properties of neonatal T cells. Phenotypically, neonatal CD4+ T cells are more immature than their adult CD45RO-/RA+ naive counterparts and they contain a subset (10-20%) of CD45RO-/RA+ CD31- cells which is very low in adults and displays some unique functional features. The activation and maturation of neonatal CD4+ T cells is particularly dependent upon the strength of CD28-mediated cosignal which dictates not only the cytokine profile released upon primary activation but also the response to IL-12. Activation of adult as well as neonatal CD4+ T cells in the context of low CD28 costimulation yields to the production of low levels of only one cytokine, i.e. IL-2. In contrast, strong CD28 costimulation supports the production of high levels of type 1 (IL-2, IFN gamma and TNF beta) and low levels of type 2 (IL-4 and IL-13) cytokines by neonatal T cells. The low levels of naive T cell-derived IL-4 are sufficient to support their development into high IL-4/IL-5 producers by an autocrine pathway. The ability of IL-12 to prime neonatal CD4+ T cells for increased production of IL-4 (in addition to IFN gamma) is observed only when CD28 cosignal is minimal. Under optimal activation conditions (i.e. with anti-CD3/B7.1 or allogenic dendritic cells) the response and the maturation of neonatal and adult naive T cells are similar. Thus the Th2 bias of neonatal immune response cannot be simply ascribed to obvious intrinsic T cell defect but rather to particular conditions of Ag presentation at priming. Unlike CD4+ T cells, neonatal CD8+ T cells strictly require exogenous IL-4 to develop into IL-4/IL-5 producers. Most importantly, anti-CD3/B7-activated neonatal CD8 T cells coexpress CD4 as well as CCR5 and CXCR4 and are susceptible to HIV-1 infection in vitro.


Subject(s)
CD4-Positive T-Lymphocytes/cytology , CD8-Positive T-Lymphocytes/cytology , Th1 Cells/cytology , Th2 Cells/cytology , Adult , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Humans , Infant, Newborn , Th1 Cells/immunology , Th2 Cells/immunology
12.
Eur J Immunol ; 27(12): 3526-31, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9464843

ABSTRACT

In addition to their capacity to induce pain, vasodilatation and fever, prostaglandins E (PGE) exert anti-inflammatory activities by inhibiting the release of pro-inflammatory cytokines by macrophages and T cells, and by increasing interleukin (IL)-10 production by macrophages. We here report that PGE2, the major arachidonic acid metabolite released by antigen-presenting cells (APC), primes naive human T cells for enhanced production of anti-inflammatory cytokines and inhibition of pro-inflammatory cytokines. Unfractionated as well as CD45RO- CD31+ sort-purified neonatal CD4 T cells acquire the capacity to produce a large spectrum of cytokines after priming with anti-CD3 and anti-CD28 monoclonal antibodies (mAb), in the absence of both APC and exogenous cytokines. PGE2 primes naive T cells in a dose-dependent fashion for production of high levels of IL-4, IL-10 and IL-13, and very low levels of IL-2, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and TNF-beta. PGE2 does not significantly increase IL-4 production in priming cultures, whereas it suppresses IL-2 and IFN-gamma. Addition of a neutralizing mAb to IL-4 receptor in primary cultures, supplemented or not with PGE2, prevents the development of IL-4-producing cells but does not abolish the effects of PGE2 on IL-10 and IL-13 as well as T helper (Th)1-associated cytokines. Addition of exogenous IL-2 in primary cultures does not alter the effects of PGE2 on naive T cell maturation. Thus PGE2 does not act by increasing IL-4 production in priming cultures, and its effects are partly IL-4 independent and largely IL-2 independent. Together with the recent demonstration that PGE2 suppresses IL-12 production, our results strongly suggest that this endogenously produced molecule may play a significant role in Th subset development and that its stable analogs may be considered for the treatment of Th1-mediated inflammatory diseases.


Subject(s)
CD4-Positive T-Lymphocytes/immunology , Cytokines/immunology , Dinoprostone/immunology , Lymphocyte Activation/immunology , Cells, Cultured , Cytokines/biosynthesis , Dinoprostone/pharmacology , Humans , Lymphocyte Activation/drug effects
13.
Int Immunol ; 8(12): 1963-70, 1996 Dec.
Article in English | MEDLINE | ID: mdl-8982781

ABSTRACT

Previous studies have shown that human Th2 cells, unlike their murine counterparts, retain the ability to produce IFN-gamma upon activation in the presence of exogenous IL-12. Here we first extended this notion by showing that Th2-like cell clones (Th2C) are also capable of inducing IL-12 production by physiological antigen-presenting cells (APC); we next showed that these cells may express several distinct cytokine profiles depending upon the activation signal and the type of APC with which they interact. We have analyzed the production of IL-4, IL-5 and IFN-gamma by Th2C stimulated by either anti-CD3 mAb or exogenous IL-2, using peripheral blood monocytes or dendritic cells (DC) as accessory cells. We found that: (I) DC but not monocytes released IL-12 and promoted IL-12-dependent IFN-gamma production upon interaction with anti-CD3- or IL-2-stimulated Th2C and (II) ligation of CD3 was required for the production of IL-4 but not of IL-5 or IFN-gamma. Thus, depending upon the type of APC with which they interacted and the mode of activation, Th2C, expressed four distinct cytokine profiles: (i) IL-4 + IL-5, in response to anti-CD3 + monocytes; (ii) IL-4, IL-5 + IFN-gamma, in response to anti-CD3 + DC; (iii) IL-5 + IFN-gamma, in response IL-2 + DC; and (iv) IL-5 alone, in response to IL-2 + monocytes. The ability of human Th2-like cells to induce IL-12 production and to release the proinflammatory cytokines IFN-gamma and IL-5 upon IL-2-driven interactions with APC may contribute to explain how local infection exacerbates Th2-mediated diseases, like bronchial asthma and atopic dermatitis.


Subject(s)
Cytokines/biosynthesis , Cytokines/classification , Dendritic Cells/immunology , Dendritic Cells/metabolism , Interleukin-12/biosynthesis , Lymphocyte Activation , Th2 Cells/immunology , Animals , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal/pharmacology , CD3 Complex/immunology , Clone Cells , Humans , Interleukin-2/pharmacology , L Cells , Mice , Th2 Cells/metabolism , Transfection
15.
Immunology ; 88(1): 110-5, 1996 May.
Article in English | MEDLINE | ID: mdl-8707335

ABSTRACT

The CD31 antigen (PECAM-1) has been reported to be a stable marker for a human CD4 T-cell subpopulation unable to produce interleukin-4 (IL-4). We show here that CD31 expression is not stable inasmuch as CD4 T-cell lines and clones derived from cell-sorted neonatal CD31+ cells lose CD31 upon repetitive cycles of stimulation and IL-2 expansion. Moreover, various cytokines (IL-1 alpha, IL-4, IL-6, transforming growth factor-beta) fail to reinduce CD31 on CD31- clones. Whereas all CD31+ CD4 T cells rapidly express high levels of the CD45RO antigen and down-regulate the L-selectin antigen after priming, CD31 disappears more slowly because only part of the cells lose CD31 expression upon each cycle of stimulation. Loss of CD31 reflects a functional maturation of CD45RO+ cells since, in a system which favours the development of Th2 effectors, IL-4 is produced by CD31- but not CD31+ effector T cells, whereas interferon-gamma is produced by both types of cells. However, CD31 is not a Th1 marker since it is not expressed on several Th1 antigen-specific clones. We conclude that CD31 is a maturation marker expressed on the great majority of naive CD45RO- CD4 T cells and on a subset of CD45RO+ CD4 T cells that are at an intermediate stage of maturation.


Subject(s)
Antigens, Differentiation, Myelomonocytic/metabolism , CD4-Positive T-Lymphocytes/immunology , Cell Adhesion Molecules/metabolism , T-Lymphocytes, Helper-Inducer/physiology , Adult , Biomarkers , CD4-Positive T-Lymphocytes/cytology , CD4-Positive T-Lymphocytes/metabolism , Cell Differentiation , Cells, Cultured , Cytokines/biosynthesis , Humans , Infant, Newborn , L-Selectin/metabolism , Leukocyte Common Antigens/metabolism , Microscopy, Fluorescence , Platelet Endothelial Cell Adhesion Molecule-1 , Th1 Cells/physiology , Th2 Cells/physiology , Tumor Necrosis Factor Receptor Superfamily, Member 7/metabolism
16.
Eur J Immunol ; 25(12): 3517-20, 1995 Dec.
Article in English | MEDLINE | ID: mdl-8566047

ABSTRACT

It was recently demonstrated that naive human and mouse CD4 T cells release low but sufficient levels of interleukin (IL)-4 at priming to support their development into IL-4 producers. To determine whether this IL-4 is produced by a minor subset of cells, freshly isolated human naive CD4 T cells were directly cloned by limiting dilution in the absence of exogenous IL-4. More than 95% of neonatal and 60% of adult naive T cells seeded in single-cell cultures could be expanded upon stimulation with anti-CD3 mAb immobilized on CD32-B7.1 L cell transfectants in the presence of IL-2. All 171 clones derived from four neonates and two adults produced IL-4 and IL-5 at generally high levels. Like the allergen-specific human Th2 clones described in the literature, these T cell clones produced little or no interferon-gamma upon stimulation via their T cell receptor/CD3 complex, whereas they released high levels of this cytokine when activated with phorbol 12-myristate 13-acetate+ionomycin. Cells cloned and expanded in the presence of anti-IL4 + anti-IL-4R mAb produced much lower levels of IL-4 and IL-5. It is concluded that almost every single naive human CD4 T cell primed and expanded in the absence of exogenous IL-4 releases sufficient autocrine IL-4 to support its clonal expansion into high IL-4/IL-5 producers.


Subject(s)
CD4-Positive T-Lymphocytes/metabolism , Interleukin-4/biosynthesis , Interleukin-5/biosynthesis , Th2 Cells/metabolism , Adult , CD4-Positive T-Lymphocytes/cytology , Cell Differentiation/immunology , Clone Cells , Fetal Blood/immunology , Humans , Infant, Newborn , Lymphocyte Activation , Th2 Cells/cytology
17.
Int Immunol ; 7(12): 1987-93, 1995 Dec.
Article in English | MEDLINE | ID: mdl-8746568

ABSTRACT

We previously reported that human naive CD4 T cells differentiate into effector cells producing type 1 (IL-2, IFN-gamma) and type 2 (IL-4, IL-5, IL-10) cytokines after priming with anti-CD3 mAb presented on irradiated CD32-transfected mouse L fibroblasts, in the absence of exogenous cytokine. Here we first show that the CD32 L fibroblasts act not only by cross-linking anti-CD3 mAb but also by providing a B7-mediated co-stimulation signal which is required for the activation of naive T cells. Using a selected anti-CD3 mAb (64.1) we next demonstrate that colligation of CD3 and CD28 with soluble mAb is sufficient to activate highly purified naive CD4 T cells for proliferation, IL-4 mRNA expression, IL-4 secretion, and maturation into IL-4- and IL-5-producing cells. Finally, we show that the intensity of B7 co-stimulation at priming markedly affects the lymphokine-producing phenotype of primed cells. Indeed, cells primed on CD32-B7 double L transfectants produce much more IL-4 and IL-5 and slightly less IFN-gamma than those primed on CD32 L cells. The enhanced IL-4/IL-5-producing capacity of cells primed on CD32-B7 L fibroblasts may be related to increased IL-4 production during priming. It is suggested that the maturation of naive T cells along the Th2 or Th1 pathway may be regulated by the level of B7 expressed on APC.


Subject(s)
B7-1 Antigen/immunology , CD4-Positive T-Lymphocytes/cytology , CD4-Positive T-Lymphocytes/immunology , Animals , Antibodies, Monoclonal/administration & dosage , CD28 Antigens/immunology , CD3 Complex/immunology , Cell Differentiation/immunology , Cytokines/biosynthesis , Humans , In Vitro Techniques , Infant, Newborn , Interleukin-4/biosynthesis , L Cells , Lymphocyte Activation , Mice , Phenotype , Receptors, IgG/genetics , Signal Transduction/immunology , Transfection
18.
Eur J Immunol ; 25(9): 2722-5, 1995 Sep.
Article in English | MEDLINE | ID: mdl-7589153

ABSTRACT

The maturation of naive CD4 T cells into interleukin (IL)-4-producing effectors was shown to require the presence of IL-4 at priming, the cellular origin of which remains unclear. We demonstrate here that naive T cells themselves release IL-4 at very low levels that are nevertheless sufficient to promote their development into Th2-like cells. This conclusion is based on three observations: (1) highly purified human naive CD4 T cells, of neonatal or adult origin, develop into Th2 effectors upon repetitive cycles of stimulation with anti-CD3 monoclonal antibody (mAb) cross-linked to CD32-B7 transfected L fibroblasts followed by IL-2 expansion; (2) IL-4 protein is readily detectable in the concentrated supernatant fluids of priming cultures performed in the presence of anti-IL-4 receptor mAb; and (3) addition of anti-IL-4 or anti-IL-4 receptor mAb at priming markedly inhibits the acquisition of IL-4- and IL-5-producing capacity while enhancing that of interferon-gamma.


Subject(s)
CD4-Positive T-Lymphocytes/immunology , Interleukin-4/biosynthesis , Th2 Cells/immunology , Adult , CD4-Positive T-Lymphocytes/cytology , Cell Differentiation/drug effects , Cells, Cultured , Humans , Interleukin-4/pharmacology , Th2 Cells/cytology
19.
J Immunol ; 153(11): 4862-71, 1994 Dec 01.
Article in English | MEDLINE | ID: mdl-7963550

ABSTRACT

Naive CD8 T cells isolated from umbilical cord blood were examined for cytokine production and for surface phenotype before and after priming with anti-CD3 mAb in the presence of selected cytokines. On stimulation with anti-CD3 immobilized on CD32-B7-transfected L cells, naive and primed CD8 T cells release high levels of IFN-gamma but no IL-2, IL-4, or IL-5. IL-12-primed cells, and to a lesser extent IL-2-primed cells, have an increased capacity to produce IFN-gamma but display the same restricted pattern of cytokine production. Cells primed in the presence of IL-4 or IL-4 + IL-2 are capable of producing high levels of IL-5 but no IL-4, and their IFN-gamma-producing capacity is much reduced. Cells primed with both IL-4 + IL-12 produce high levels of IL-5 and IFN-gamma but no IL-4. IL-4-producing CD8 T cells are obtained only if IL-4- or IL-4 + IL-2-primed cells are subjected to a second cycle of activation in the presence of IL-2; restimulation of IL-4 + IL-12-primed cells under identical conditions fails to generate IL-4-producing cells but leads to the development of high IFN-gamma and IL-5 producers. Thus, unlike in CD4 T cells, IL-12 completely inhibits IL-4-induced capacity of CD8 T cells to produce IL-4. However, IL-4 and IL-12 synergize to promote the expression of IL-5. Regardless of the priming conditions, primed CD8 T cells are CD45ROhigh/RA-, CD31high and more than 50% are CD4+/CD8+; activated naive or primed CD8 T cells do not express CD40 ligand.


Subject(s)
CD8-Positive T-Lymphocytes/immunology , Cytokines/biosynthesis , CD3 Complex/immunology , Cell Differentiation/immunology , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Fetal Blood/cytology , Flow Cytometry , Humans , Immunophenotyping , Infant, Newborn , Interferon-gamma/biosynthesis , Interleukin-12/biosynthesis , Interleukin-2/biosynthesis , Interleukin-4/biosynthesis , Interleukin-5/biosynthesis , Radioimmunoassay
20.
J Clin Invest ; 94(4): 1352-8, 1994 Oct.
Article in English | MEDLINE | ID: mdl-7929809

ABSTRACT

It is now recognized that IL-12 plays a predominant role in protective immunity against intracellular pathogens by promoting the development of T helper type 1 (Th1) responses. We here report the unexpected observations that IL-12 exerts differential effects on the maturation of "native" human CD4 T cells isolated from umbilical cord blood or from the blood of healthy adults. After priming in the presence of IL-12, naive cells of adult donors, defined as CD45R0- CD4+ T cells, acquire a Th1 phenotype whereas neonatal cells develop into effector cells producing high levels of IL-4 in addition to IFN-gamma. This effect of IL-12 on neonatal T cells is direct inasmuch as it is observed on highly purified CD4 T cells, however, it is not inhibited by CD8 T cells and natural killer cells. Unstimulated neonatal T cells which have been preincubated with IL-12 before the priming behave like adult T cells and acquire a Th1 phenotype after stimulation in the presence of IL-12. Given that IL-4 is a potent antagonist of Th1 responses, the finding that IL-12 promotes the maturation of neonatal T cells into IL-4 producers may explain the increased susceptibility of neonates to intracellular pathogens and should be taken into account for the development of vaccines to be used in the perinatal period.


Subject(s)
CD4-Positive T-Lymphocytes/immunology , Interferon-gamma/biosynthesis , Interleukin-12/pharmacology , Interleukin-4/biosynthesis , Adult , CD4-Positive T-Lymphocytes/drug effects , Cells, Cultured , Humans , Infant, Newborn , Interleukin-1/pharmacology , Lymphocyte Activation , Recombinant Proteins/pharmacology , Tumor Necrosis Factor-alpha/pharmacology , Umbilical Cord/cytology
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