Immature Langerhans Cells Research Articles

Antigen processing in populations of mature murine dendritic cells is caused by subsets of incompletely matured cells.

Immature dendritic cells (DC), such as freshly isolated Langerhans cells (LC), are excellent at processing native protein Ag. During short term culture they shut off MHC class II synthesis and down-regulate their processing capacity. They retain, however, the MHC/peptide complexes, up-regulate adhesion and costimulatory molecules, and acquire the ability to sensitize T cells. Two reports describing substantial processing activity in populations of mature DC prompted us to undertake an extensive comparative study of the Ag-processing capacities of immature vs mature DC. We used a panel of 17 peptide-specific T cell hybridomas restricted by six different MHC class II molecules: I-Ab, I-A(d), I-E(d), hybrid I-A beta dE alpha, I-Ak, and I-Ek. Side by side comparisons revealed in all cases that freshly isolated LC were superior to cultured mature LC in their ability to process native proteins. With some hybridomas, however, we found a considerable degree of processing by populations of cultured LC at high doses of Ag or Ag-presenting cells. This activity, however, did not correlate with the MHC haplotype. Direct comparison over wide ranges of DC doses or Ag doses showed that it was always less than that of corresponding fresh immature LC. Immunoperoxidase staining of cytospins and flow cytometry with mAb In1 disclosed a small (20% maximum) subset of cultured LC expressing the MHC class II-associated invariant chain, indicating ongoing biosynthesis of this molecule and, thus, incomplete maturation of these LC. Therefore, the residual processing activity observed in populations of mature DC may be explained by small subpopulations of incompletely matured DC.

Postnatal Development of the Vaginal Epithelium in the Mongolian Gerbil (<i>Meriones unguiculatus</i>)

This study examines the normal postnatal (p.n.) development of the Mongolian gerbil's vagina from birth to the age when consistent cyclic changes of the estrus occur. At birth, the Müllerian vagina has a lumen and is lined by a pseudostratified or 2- to 3-layered epithelium. The cells contain large amounts of glycogen. The sinus vagina forms a solid plate and contains hardly any glycogen. During the first few p.n. days the cell layers separate, and the newly formed lumen becomes continuous with the Müllerian vagina. Only a vaginal closure membrane persists. Between p.n. days 10 and 15 the epithelium increases in height reaching 2-5 cell layers. The amount of glycogen diminishes, but in the luminal cells the first mucous granules develop. Leukocyte diapedesis starts, and immature Langerhans cells become discernible. Between p.n. days 18 and 30 the apical cells become mucified, and some shedding of these cells begins. Between p.n. days 36 and 45 bundles of keratin filaments appear in many of the luminal mucous cells intermingled with the mucous granules. In the middle layers keratin filaments, keratohyalin granules and membrane-coating granules indicate the beginning of the keratinization process. Diapedesis of leukocytes may lead to cell aggregations below top layers. By day 50 the shedding of the mucous cell layers reveals the underlying keratinized cell layers which then become exfoliated. The first steps of the estrous cycle show a heterogeneous pattern. At about p.n. day 60 the cycle becomes established.

Migration and maturation of Langerhans cells in squamous metaplasia of the rat trachea induced by vitamin A deficiency.

The migration and maturation of Langerhans cells (LCs) in rat tracheal squamous metaplasia due to vitamin A deficiency were investigated immunohistochemically and electron microscopically. In the early stage of metaplasia, i.e. basal cell hyperplasia, no LCs with Birbeck granules (BGs) could be found, but there were desmosome-free cells which had the morphological characteristic of immature LCs. They were clearly different from inflammatory cells such as macrophages and lymphocytes, and were, therefore, considered to be precursors of LCs. In the stage of stratification, small numbers of Ia- and protein kinase C type II (PKCII)-positive cells were recognized. Ultrastructurally they were immature LCs with ovoid nuclei, many free ribosomes and few dendrites. The cytoplasm was dark and a few BGs and atypical granules (AGs) could be seen in the Golgi area. In the early stage of cornification, LCs with partially intended nuclei, prominent nucleoli and well-developed Golgi complexes were found. There were many BGs and AGs and structures transitional between them in the Golgi areas. In epithelium showing mature squamous metaplasia, many Ia- and PKCII-positive dendritic cells could be seen. Most of these were typical mature LCs with lobulated nuclei, clear cytoplasm and prominent dendritic processes. The number of BGs and AGs were fewer than in the LCs found in the early stage of cornification, and these granules were distributed throughout the cytoplasm. In the final stage, where the basal cells had differentiated into a flatter epithelium, few LCs could be seen.(ABSTRACT TRUNCATED AT 250 WORDS)

Maturational steps of bone marrow-derived dendritic murine epidermal cells. Phenotypic and functional studies on Langerhans cells and Thy-1+ dendritic epidermal cells in the perinatal period.

The adult murine epidermis harbors two separate CD45+ bone marrow (BM)-derived dendritic cell systems, i.e., Ia+, ADPase+, Thy-1-, CD3- Langerhans cells (LC) and Ia-, ADPase-, Thy-1+, CD3+ dendritic epidermal T cells (DETC). To clarify whether the maturation of these cells from their ill-defined precursors is already accomplished before their entry into the epidermis or, alternatively, whether a specific epidermal milieu is required for the expression of their antigenic determinants, we studied the ontogeny of CD45+ epidermal cells (EC). In the fetal life, there exists a considerable number of CD45+, Ia-, ADPase+ dendritic epidermal cells. When cultured, these cells become Ia+ and, in parallel, acquire the potential of stimulating allogeneic T cell proliferation. These results imply that CD45+, Ia-, ADPase+ fetal dendritic epidermal cells are immature LC precursors and suggest that the epidermis plays a decisive role in LC maturation. The day 17 fetal epidermis also contains a small population of CD45+, Thy-1+, ADPase-, CD3- round cells. Over the course of 2 to 3 wk, they are slowly replaced by an ever increasing number of round and, finally, dendritic CD45+, Thy-1+, CD3+ EC. Thus, CD45+, Thy-1+, ADPase-, CD3- fetal EC may either be DETC precursors or, alternatively, may represent a distinctive cell system of unknown maturation potential. According to this latter theory, these cells would be eventually outnumbered by newly immigrating CD45+, Thy-1+, CD3+ T cells--the actual DETC.

Isolation and characterization of a cDNA and gene coding for a fourth CD1 molecule.

The CD1 locus encodes a family of major histocompatibility complex (MHC) antigen-like glycoproteins which associate with beta 2-microglobulin and are expressed on immature thymocytes and Langerhans cells. Three CD1 molecules have been identified by monoclonal antibodies and molecular cloning: CD1a, -b, and -c. We have isolated a cDNA coding for a fourth CD1 molecule from a human thymocyte library and termed this molecule CD1d. Reported here are the complete nucleotide sequence and genomic organization of CD1d. They predict that this molecule is related to the previously identified CD1a, -b, and -c molecules and to MHC class I molecules, with three external domains, a transmembrane domain, and a short cytoplasmic tail. The sequence of CD1d is the most divergent among the CD1 molecules in the membrane-distal alpha 1 and alpha 2 domains and in the 5' untranslated region. In contrast, all four CD1 molecules are highly homologous in the membrane-proximal alpha 3 domain, which is likely involved in beta 2-microglobulin binding. A comparison of CD1 and MHC class I sequences suggests that these molecules each evolved to interact with a distinct set of cell surface proteins.

ラット口蓋粘膜のランゲルハンス細胞の加齢的変化に関する研究

The function of Langerhans cells (LCs) is unknown for a long time. Since the antigen-presenting function of LCs was demonstrated, the study of LCs has been increased. Although the existence of LCs has been found in oral mucosa of a human being and animals, the previous researches on age change of numerical distribution and morphology of LCs are very few. In order to understand the oral immunity, the palatal mucosa of Wistar rats were used in this study from lst day to 1 year after birth. The changes of morphology and numerical distribution of LCs were observed as follows : The development of large process of LCs was found under the Hematoxylin-Eosin stain except of the slender dendritic ones. The number of LCs has reached the peak at 1 month after birth since immature LCs of palatal mucosa were found at 1st day after birth, simultaneously, the difference of numerical distribution of LCs between the hard and soft palate became more evident at 1 month after birth. Therefore, it was considered that the development and environment of oral cavity rapidly changed after weaning period. Thereafter, an increase or a decrease in number of LCs distribution was observed and a gradual decline in the numerical distribution of LCs was also shown with age as the regressive changes of oral mucosa. Thus, the role of immunological function of oral mucosa might be lower with advancing age.

Development of ATPase-positive, immature Langerhans cells in the fetal mouse epidermis and their maturation during the early postnatal period.

The development and maturation of Langerhans cells during the differentiation of skin was studied in mice from fetal day 13 to adult using 3 indices: ATPase activity: ultrastructure; and quantitative evaluation of the cell population. ATPase-positive Langerhans cells appeared in the epidermis at first at fetal day 16, and they increased in number in the differentiating epidermis during the late fetal period. The earliest appearance of Birbeck granules was at postnatal day 4. Cored tubules were also formed in the Langerhans cells in the dermis at around the same age. The cells containing Birbeck granules or cored tubules are considered to be mature Langerhans cells. In the Langerhans-cell lineage, those cells in the epidermis at stages earlier than postnatal day 4 and not yet containing specific organelles are considered to be immature Langerhans cells. These immature Langerhans cells can be identified ultrastructurally in the epidermis at fetal day 16, coinciding with the appearance of ATPase-positive cells. The increase in the number of immature Langerhans cells during the perinatal period was shown by quantitative analysis of nuclear density and relative Langerhans-cell area on the electron micrographs. It is concluded that ATPase is a marker of the Langerhans-cell lineage from the early development stages, while Birbeck granules and cored tubules are markers that identify mature Langerhans cells in electron micrographs.

Biochemistry and Biology of the Langerhans Cell

Langerhans cells mainly present in the normal epidermis and are thought to be identical to the proliferating cells in the lesions of histiocytosis-X. The Langerhans cell is positive not only for ATPase, esterase, and acid phosphatase, but also for S-100 protein. Since these findings are similar but distinct from those of monocytes and macrophages, the Langerhans cell may be a cell line independent of the monocyte-macrophage system, designated as T-zone histiocytes. Immunologically, the Langerhans cell has Fc and C3 receptors and Ia-like antigen. Thus, the Langerhans cell may be a member of the immune network, as antigen-presenting cells, in the epidermis. One hypothesis is that Langerhans cells activated by external stimuli induce a local immune reaction by activation and proliferation of T-cells, with the interaction of keratinocytes and macrophages. Because of cytochemical and immunologic similarities between Langerhans cells and histiocytosis-X cells, histiocytosis-X may be a proliferative disorder of immature or neoplastic Langerhans cells.

Ontogeny of Ia-Positive and Thy-1-Positive Leukocytes of Murine Epidermis

Murine epidermis harbors 2 populations of dendritic leukocytes: Langerhans cells (LC) and Thy-1-positive dendritic epidermal cells (Thy-1 +DEC). In the adult mouse these cell types are morphologically distinct and display highly characteristic phenotypes. LC bear Ia-antigens and a group of markers typical for mononuclear phagocytes: Fc- and C3bi-receptors, macrophage-specific antigen F4/80, and membrane ATPase. Thy-1 +DEC, in contrast, lack these markers but express high levels of Thy-1 and asialo-GM1 (asGM1) antigen. Since LC and Thy-1 +DEC share a common origin from the bone marrow we expected to gain insight into their relationship by studying their ontogenetic development. Epidermal sheets from fetal and newborn C3H/He and C57B1/6 mice obtained at defined ages from day 17 of gestation up to day 30 of postnatal life were monitored for the emergence of the above-mentioned markers for LC and Thy-1 +DEC. In double-labeling experiments LC markers were first detected by visualizing the monoclonal antibodies by a sensitive triple-layer rhodamine-immunofluorescence technique; in a second step, after appropriate blocking procedures, Thy-1 and asGM1 antigens were demonstrated by direct and indirect immunofluorescence. We found that in fetal epidermis, only few cells expressed either Thy-1 or Ia (4 and 1 cells/mm2, respectively, on day 18 of gestation). The bulk of Thy-1 +DEC and Ia +EC appeared only after birth. Adult proportions of Thy-1 +DEC and Ia +EC were reached at around 1 month of postnatal life. In contrast, all the other LC markers were expressed on a substantial number of fetal dendritic cells (280 cells/mm2 on day 18 of gestation), indicating the presence of phenotypically immature Ia-negative LC in fetal epidermis. By day 4 of postnatal life all F4/80 +EC and ATPase +EC (i.e., LC) had acquired Ia-antigens. Surprisingly, LC also bore asGM1 antigens, which in the adult epidermis are strictly confined to Thy-1 +DEC, up to day 5 of postnatal life. Thus, LC in fetal and early newborn epidermis are not yet fully differentiated. As they differentiate, they acquire Ia antigens and lose asGM1 antigens. In contrast, a phenotypically immature Thy-1 +DEC population could not be traced with the markers used. Thy-1 +DEC appear to be characterized by a stable phenotype (Thy-1+/asGM1+) throughout their lifetime.

Immunocytochemical demonstration of S-100 protein-containing dendritic cells in the normal and inflamed human gingiva.

Langerhans cells functioning as epidermal macrophages are known to contain S-100 protein, a peculiar brain-derived protein. The present study demonstrates the occurrence and ultrastructure of S-100-immunoreactive dendritic cells in normal and inflamed gingiva in human subjects. A few S-100-immunopositive cells were found scattered in the normal gingival epithelium, but these greatly increased in number in the inflamed gingiva. By electron microscopical observation, the S-100-immunopositive cells in the normal gingiva were confirmed as containing Birbeck granules characteristic of Langerhans cells, while the cells multiplying in the inflamed gingiva corresponded to indeterminate dendritic cells, which have been regarded as immature Langerhans cells in the skin. The close juxtaposition of lymphocytes to the S-100-immunopositive indeterminate dendritic cells was frequently recognized in the inflamed gingiva, a finding which suggests that these cells, and not the typical Langerhans cells, may function as antigen-presenting cells.

Demonstration of OKT 6—reactive cells in mycosis fungoides

Infiltrates of five cases of mycosis fungoides (MF) were studied for the presence of cells reactive with a monoclonal antibody, OKT 6, which detects an antigen present on relatively immature thymocytes and Langerhans cells. In situ immunohistochemical staining was used for their demonstration. OKT 6-reactive cells formed a definite component of the dermal infiltrates of all patients examined. In three patients who showed numerous Pautrier microabscesses on routine histologic examination, OKT 6-positive cells were found to form a component of these abscesses. OKT 6-reactive cells have also recently been shown to be present in varying numbers in the dermal infiltrates of large plaque (atrophic) parapsoriasis (LPAP), a condition which may terminate in MF. The significance of these findings is discussed.