Spleen dendritic cells exhibit altered morphology and increased allostimulatory capacity after short-term culture* (original) (raw)
Immunology, 1996
We compared the capacity of mature dendritic cells (DC) from lymph nodes and maturing DC from spleens in their capacity to stimulate responses to the small hapten picryl sulphonic acid (PIC) and to the same hapten conjugated to ovalbumin (PIC-OVA) and requiring processing. Surface expression of major histocompatibility complex (MHC) class II molecules, which are upregulated during maturation of splenic DC, were studied as an independent marker of maturation. Freshly isolated lymph node DC had a veiled appearance and high levels of class II expression. DC separated from suspensions of spleen cells expressed the DC-specific marker NLDC-145, but were small, had low levels of MHC class II molecules and expressed stem cell antigen. Those DC from spleen cells cultured for 24 and 48 hr showed the development of typical veiled DC morphology and high class II expression. Lymph node DC stimulated high levels of primary T-cell proliferation to PIC, but failed to stimulate primary responses to PIC-OVA. Splenic DC isolated immediately failed to stimulate primary responses to either antigen. More mature spleen DC stimulated responses both to PIC and PIC-OVA. Surprisingly, development of the capacity to stimulate responses to PIC preceded that of stimulating PIC-OVA responses. The capacity of the DC to process and present PIC-OVA was maintained during the culture period. The results indicate that both the form of the antigen and the source and maturity of the DC are critical in determining the responses stimulated in T lymphocytes.
The Dendritic Cell Populations of Mouse Lymph Nodes
The Journal of Immunology, 2001
The dendritic cells (DC) of mouse lymph nodes (LN) were isolated, analyzed for surface markers, and compared with those of spleen. Low to moderate staining of LN DC for CD4 and low staining for CD8 was shown to be attributable to pickup of these markers from T cells. Excluding this artifact, five LN DC subsets could be delineated. They included the three populations found in spleen (CD4 ؉ 8 ؊ DEC-205 ؊ , CD4 ؊ 8 ؊ DEC-205 ؊ , CD4 ؊ 8 ؉ DEC-205 ؉ ), although the CD4-expressing DC were of low incidence. LN DC included two additional populations, characterized by relatively low expression of CD8 but moderate or high expression of DEC-205. Both appeared among the DC migrating out of skin into LN, but only one was restricted to skin-draining LN and was identified as the mature form of epidermal Langerhans cells (LC). The putative LC-derived DC displayed the following properties: large size; high levels of class II MHC, which persisted to some extent even in CIITA null mice; expression of very high levels of DEC-205 and of CD40; expression of many myeloid surface markers; and no expression of CD4 and only low to moderate expression of CD8. The putative LC-derived DC among skin emigrants and in LN also showed strong intracellular staining of langerin.
Langerhans Cells and Extra-Epidermal Dendritic Cells
Scandinavian Journal of Immunology, 1985
S-HM) protein was demonstrated in the cytoplasm of dendritic cells (DCs) in normal and pathologic lymphoid tissues and epidermis in man and several other species. The presence of S-U)() protein served to distinguish these cells from other mononuclear cells, most importantly from those of maerophage/histiocyte lineage. Fractionation procedures to isolate and enrich suspensions of DCs were eoupled with immunoeytochemical techniques to identify S-KHI-positive cells, Langerhans cells in the epidermis and in aural cholesteatomata and nodal, splenie, and thymie interdigitating cells were S-100-positive. Lymph node and splenie foilicular dendrilie cells (except in rats) were negative, indieating that this DC may be a separate cell type.
Journal of Investigative Dermatology, 2001
The dermis harbors a true dendritic cell population that could elicit primary allogeneic T cell responses in vitro and contact hypersensitivity reactions in vivo. The origin of dermal dendritic cells remains poorly understood, however. In this study, we analyzed the fate of monocytes or monocyte-derived dendritic cells in a dermal equivalent. Freshly isolated monocytes or monocytes cultured for 6 d with either GM-CSF/IL-4 or GM-CSF/IL-4/TGF-b1 (TGF-DC) were seeded in a collagen solution with normal human ®broblasts. The lattices were cultured for 7± 14 d in the presence, or absence, of the exogenous cytokines, before phenotypic and functional studies were performed. Supply of exogenous cytokines allows the appearance of typical CD1a + /CD14 ± / CD68 low dendritic cells with signi®cant allostimulatory property, regardless of the cell type incorporated into the lattices. In cytokine-free conditions, monocytes and GM-CSF/IL-4-derived dendritic cells give rise to a CD1a ± /CD14 + /CD68 high monocyte/ macrophage population with no allostimulatory property. When incorporated into the lattices in the absence of exogenous cytokines the TGF-DC express few CD68 and FXIIIa. Interestingly, these cells do not all convert into the CD14 + /CD1a ± population. Indeed, a small HLA-DR + /CD1a + /CD14 ± subset was consistently found, which represents about one-third of the HLA-DR + cells. Moreover, TGF-DC recovered from the lattices after culture without cytokines do display a signi®cant allostimulatory function. Thus, in the absence of exogenous cytokines, only Langerhans-cell-like dendritic cells can retain the typical dendritic cell features when inserted in a dermal environment. Taken together, these results may provide evidence supporting an epidermal origin of dermal dendritic cells.
Most lymphoid organ dendritic cell types are phenotypically and functionally immature
2003
Dendritic cells (DCs) have been thought to follow a life history, typified by Langerhans cells (LCs), with 2 major developmental stages: an immature stage that captures antigens in the periphery and a mature stage that presents those antigens in the lymphoid organs. However, a systematic assessment of the maturity of lymphoid organ DCs has been lacking. We have analyzed the maturity of the DC types found in the steady state in the spleen, lymph nodes (LNs), and thymus.
Factors determining the spontaneous activation of splenic dendritic cells in culture
Innate Immun, 2011
Dendritic cells (DCs) serve as a link between the innate and adaptive immune systems. The activation state of DCs is crucial in this role. However, when DCs are isolated from lymphoid tissues, purified and placed in culture they undergo 'spontaneous' activation. The basis of this was explored, using up-regulation of DC surface MHC II, CD40, CD80 and CD86 as indicators of DC activation. No evidence was found for DC damage during isolation or for microbial products causing the activation. The culture activation of spleen DCs differed from that of Langerhans cells when released from E-cadherin-mediated adhesions, since E-cadherin was not detected and activation still occurred with b-catenin null DCs. Much of the activation could be attributed to DC-DC interactions. Although increases in surface MHC II levels occurred under all culture conditions tested, the increase in expression of CD40, CD80 and CD86 was much less under culture conditions where such interactions were minimised. DC-to-DC contact under the artificial conditions of high DC concentration in culture induced the production of soluble factors and these, in turn, induced the up-regulation of co-stimulatory molecules on the DC surface.
Journal of Investigative Dermatology, 1989
Dendritic epidermal T cells (DETC) are CD45+, Thy-l+, CD5-, CD8-, CD4murine lymphocytes that express surface-bound CD3 antigens associated with T cell receptor y/S heterodimers. Using epidermal cells greatly enriched for DETC and depleted of Langerhans cells, we found that DETC have growth requirements quite different from those of accessory cell-depleted lymph node and splenic T cells. Although the latter cells strongly proliferate in response to phorbol myristate acetate (PMA) + ionomycin, DETC, when exposed to interleukin-1 (IL-l), interleukin-3 (IL-3), concanavalin A (ConA), PMA, and ionomycin used either alone or in combination, do not exhibit significant mitotic activity. Recombinant interleukin 2 (rIL-2), albeit ineffective by itself, leads to vigorous proliferation of DETC when used with either ConA or PMA + ionomycin + ILl. In contrast, the combination of PMA and recombinant interleukin-4 (rIL-4), which triggers growth of lym h node T cells, does not induce proliferation of DETC. A though a portion of P proliferating DETC expressed CD8 antigens, essentially none bore detectable amounts of surface-bound CD4 or CD5 antigens, or both. Continuing stimulation of primary DETC cultures with lectin/lymphokine-rich media results in the propagation of cells with the essential phenotypic features of resident DETC.] Invest Dermatol92:763-768, 1989 I n 1983, the authors and others [1,2] described a hitherto unrecognized cell system within the murine epidermis. These cells are bone marrow-derived [1,3-51, display a highly dendritic morphology in situ, and bear large amounts of the Thy-l alloantigen on their surface [1,2]. They are clearly distinct from all other epidermal cells (EC), i.e., keratinocytes (KC), Langerhans cells (LC), and melanocytes, and have been referred to as Thy-l + dendritic e idermal cells (Thy-l+DEC) [1,2,5,6]. In single EC suspensions, t R ese cells exhibit a round shape and comprise only l-2% of all EC [1,2]. Although Thy-l +DEC are different from most peripheral T cells as evidenced by their lack of CD5, CD4, and CD8 antigens [6], compelling evidence exists that they belong to the T cell system: they are numerically reduced in athymic mice [ 11; they proliferate in response to concanavalin A (ConA) + interleukin 2 (IL-2) [7]; and they uniformly express surface-bound CD3 antigens [8]. These CD3 antigens are predominantly, if not exclusively, associated with Manuscript
Journal of Immunological Methods, 1998
We have developed a direct immunocytochemical technique to identify cytokine and chemokine production in epidermal Ž . Ž . Langerhans cells LC and in vitro derived CD14y , CD1a q , CD83q , CD40q dendritic cells DC at the single cell level. Formaldehyde fixation combined with saponin permeabilization preserved cellular morphology and generated a characteristic juxtanuclear staining signal due to the accumulation of cytokine to the Golgi organelle. This approach was used for the assessment of TNF-a, IL-6, IL-8, IL-10, IL-12, GM-CSF, MIP-1a , MIP-1b and RANTES producing cells. In contrast, a diffuse cytoplasmic staining was evident for IL-1ra, IL-1a and IL-1b production. IL-1ra and IL-1a were expressed in 10-25% of unstimulated cultured cells, while all the other cytokines were undetectable. IL-1ra, IL-1a and IL-1b were also the dominating cytokines, expressed in up to 85% of the DC, after 3 h of LPS stimulation. A significantly Ž . lower number of cells 0-5% synthesized TNF-a, IL-6, IL-10, IL-12 and GM-CSF. The incidence of chemokine producing Ž . cells IL-8, RANTES, MIP-1a , MIP-1b peaked 10 h after LPS stimulation in up to 60% of the DC. Both immature CD83y and mature CD83q DC as well as LC had a similar cytokine production pattern. Thus, in comparison to monocytes, LPS stimulation of DC generated a lower incidence of TNF-a, IL-6, IL-10 and IL-12 producing cells while IL-1 was expressed in a comparable number of cells. q
Immunophenotypic analysis of normal human dendritic cells isolated from epidermis and dermis
International Journal of Dermatology, 1998
Anatomy and Histology and the Background The skin immune system comprises two types of dendritic cells, i.e. CD1a-Dermatology Clinic, University of positive Langerhans cells in the epidermis and CD36-positive dendritic macrophages in the Florence, Florence, Italy dermis. Dendritic cells can migrate from skin explants into a culture medium. Methods We have examined the morphology and immunophenotype of the dendritic cells
Dendritic cells in the rat spleen follicles
Cell and Tissue Research, 1978
Follicles of peripheral lymphoid organs (rat) contain a type of nonlymphoid cell which is capable of arresting antigen-antibody complexes at the cell surface. These so-called dendritic cells can be visualized in immunized rats by staining antigen-antibody complexes with immunohistoperoxidase techniques.
Dendritic Cells from Peyer's Patch and Spleen Induce Different T Helper Cell Responses
Journal of Interferon & Cytokine Research, 1998
The role of antigen-presenting cells (APC) in regulating the balance of T helper type 1 (Thl) and T helper type 2 (Th2) responses and cytokine production is unclear. Dendritic cells (DC), the most potent APC for naive T cell activation, were found to regulate Thl and Th2 cytokine profiles in a manner dependent on their tissue of origin. Using whole tissues or purified cell mixtures, spleen (systemic) DC were found to induce mainly Thl cytokines, and Peyer's patch (mucosal) DC were found to induce predominantly Th2 cytokines. Spleen DC induced high levels of interferon-y (IFN-y) or interleukin-2 (IL-2) or both, and Peyer's patch DC induced IL-4 or IL-6 or both in spleen and Peyer's patch T cells, allogeneic mixed leukocyte reactions, or antigen-specific ThO clones. These data suggest that the tissue of origin of DC has a significant impact on subsequent T cell development. produce IL-12 and direct the development of Thl cells from naive CD4+ T cells. J. Immunol. 154, 5071-5079.
Langerhans cells cross-present antigen derived from skin
Proceedings of The National Academy of Sciences, 2006
, the DCs of the epidermis. Therefore, we investigated this issue in detail. Isolated murine LCs were able to cross-present soluble ovalbumin protein on MHC-class I molecules to antigen-specific CD8 ؉ T cells, albeit less potently than the CD8 ؉ DC subsets from spleen. Furthermore, LCs cross-presented cell-associated ovalbumin peptide and protein expressed by neighboring keratinocytes. Use of transporter associated with antigen processing (TAP-1)-deficient mice suggested a TAP-dependent pathway.
Journal of Experimental Medicine, 1993
A procedure has been developed to isolate dendritic cells to a high degree of purity from fresh blood. Prior enrichment methods have relied upon an initial 1-2-d culture period. Purified fresh isolates lack the characteristic morphology, phenotype, and immunostimulatory function of dendritic cells. The purified cells have the appearance of medium sized lymphocytes and express substantial levels of CD4, but lack the T cell molecules CD3, CD8, and T cell receptor. When placed in culture, the cells mature in a manner resembling the previously described, cytokine-dependent maturation of epidermal dendritic cells (Langerhans cells). The cells enlarge and exhibit many cell processes, express much higher levels of major histocompatibility complex class II and a panel of accessory molecules for T cell activation, and become potent stimulators of the mixed leukocyte reaction. Among the many changes during this maturation process are a fall in CD4 and the appearance of high levels of B7/BB1, ...
Origin of follicular dendritic cell in the chicken spleen
Cell and Tissue Research, 2006
The ellipsoid-associated cell (EAC) is a bloodborne phagocytic cell, residing in the antigen trapping zone of the chicken spleen. Binding and endocytosis of βGalactosidase (βGal) are independent from the Fc and complement receptors, because sulfated polysaccharides, in a concentration manner, inhibit the bacterial antigen uptake. The βGalpositive cells migrate to the periarterial lymphatic sheath (PALS), the preexisting germinal centers (GC), and form clusters with B-and T-cells. βGal, E5G12 double positive cells on the surface of the ellipsoid and in the PALS, GC and clusters prove that the EACs carry the enzyme. The EAC and the follicular dendritic cell (FDC) express, 68.2 and E5G12 and, 74.3 and E5G12, antigens, respectively. During migration the cessation of 68.2 and expression of 74.3 indicate the differentiation of EAC to FDC. By day 14 the clusters had disappeared, and in several GC the presence of double positive cells (74.3 and βGal; E5G12 and βGal) showed that the clusters had developed to GC. The presence of βGal + cells in the PALS, where interdigitating dendritic cells (IDC) cooperate with the T-cells, suggests that in the spleen alternate routes exist for the EAC differentiation to FDC: EAC to FDC: βGal-loaded cells in the preexisting GC; and EAC through IDC to FDC: βGal + EAC in the PALS and clusters. The EAC-FDC axis works exclusively inside the spleen; therefore; this system may be operated in pneumococcus infection.