Large Granular Cells Research Articles

Endoscopic resection of a large granular cell tumor of the esophagus.

Endoscopic resection of a large granular cell tumor of the esophagus.

Granulocyte-macrophage colony-stimulating factor enhances interleukin-1β stimulated histamine release in the preovulatory rat ovary

The existence of immune cells including macrophages and mast cells in rat ovary implies that various cytokines from these cells may play a role in ovarian functions. The aim of the present study was to investigate whether granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-1β are capable of stimulating histamine release and steroidogenesis in rat ovary, and to determine the sites of histamine production in the ovary. Histamine release from preovulatory ovarian tissues was stimulated in a dose-dependent manner at 3–30 ng/ml of GM-CSF in the presence of interleukin-1β (10 ng/ml). However, treatment with GM-CSF and interleukin-1β did not cause any significant change in the levels of ovarian steroids. Intense staining of histidine decarboxylase in the ovary was immunohistochemically detected in large granular cells on the morning of the pro-oestrus day. These results indicate that GM-CSF may be involved in the regulation of ovarian histamine secretion in mast cells partially by enhancing interleukin-1β-induced histamine release in the process of ovulation.

Tumor-like lesions in the mantle of the mussel Modiolus difficilis from the Sea of Japan.

Two inner growths in the mantle beneath the epithelium were found in 1 of 1000 mussels Modiolus difficilis from Amursky Bay, Sea of Japan, within the city precincts of Vladivostok. Both growths were about 2000 microns in maximal diameter in section and elevated slightly above the mantle surface. The mantle epithelium near the growths formed deep invaginations, and clusters of mucous cells were numerous beneath the epithelium. Histological and histochemical methods were employed. Two different kinds of growth were revealed. The off-white growth consisted of cells with thin granular or vesicular cytoplasm containing glucosaminoglycans, proteins and a small amount of neutral polysaccharides. Growth cells were pure white in color after treatment of preparations with 1% H2SO4 and differed markedly from the mantle cells. The yellow growth consisted of large granular cells with neutral polysaccharides and proteins. Although growths were composed of different kinds of cells, they seemed to be derived from subepithelial mucous cells. This was supported by histological and histochemical staining reactions of some tumor and mantle epithelial cells. Mitotic indices (MI) of growths and subepithelial mucous cells were zero, MI of ciliated mantle epithelium reached 0.07%. The lesions were areas of strongly altered mucous cells of mantle epithelium and were non-neoplastic.

IL-2Rα on One Cell Can Present IL-2 to IL-2Rβ/γc on Another Cell to Augment IL-2 Signaling

AbstractIL-2Rα augments IL-2 signaling. Although this is generally believed to occur only when the three known components of IL-2R are associated within a single cell membrane, we demonstrate here an intercellular interaction. Cocultivation of cells individually expressing chimerae incorporating the extracellular domain of IL-2Rα alone with cells expressing chimerae of IL-2Rβ alone permitted IL-2 dose-dependent oligomerization of the chimerae. Likewise, native IL-2Rα-bearing cells augmented the IL-2 proliferative response of ex vivo large granular lymphocytic leukemia cells expressing IL-2Rβ/γc but lacking IL-2Rα. In both cases, the response was inhibitable by an Ab to IL-2Rα. Intercellular augmentation of cytokine effects, acting in trans, has important implications for biology and medicine.

Development of a monocyte/macrophage-like cell line, RTS11, from rainbow trout spleen

A new rainbow trout cell line, RTS11, arose spontaneously from a long-term haemopoietic culture of spleen. For routine subculture, RTS11 requires 20–30% fetal bovine serum, a high cell density, and conditioned medium from previous cultures. Through successive passages, RTS11 cultures maintain a balance between small, non-adherent cells that form the majority of cells (referred to as round cells) and a few larger, granular cells that are either adherent or in suspension surrounded by clusters of round cells. The large granular cells appear to be macrophages, and are referred to as macrophage-like cells, because they are phagocytic, take up DiI-acetylated low density lipoprotein and acridine orange and stain for non-specific esterase, whereas the round cells appear to be at an earlier stage of macrophage development. Medium conditioned by RTS11 also contains lysozyme-like activity, an additional indication that they are of the macrophage lineage. In proliferation assays, the non-adherent cells are stimulated strongly by RTS11 conditioned medium and by lipopolysaccharide. Rainbow trout head kidney leucocytes also show a positive proliferative response to RTS11 conditioned medium. Therefore, RTS11 cells appear to both produce and respond to an autocrine growth factor(s).

Elevation of the Serum Fas Ligand in Patients With Hemophagocytic Syndrome and Diamond-Blackfan Anemia

Fas ligand (FasL) is a membrane protein that is expressed in activated T cells and natural killer cells. FasL binds to Fas on target cells and induces apoptosis. There exists a soluble form of FasL (sFasL), and sFasL also induces apoptosis of Fas-bearing cells. The serum sFasL concentrations were reported to be elevated in patients with large granular lymphocytic leukemia and natural killer cell lymphoma. In this study, we have measured serum sFasL concentrations in other hematological disorders, including severe aplastic anemia (SAA), hemophagocytic lymphohistiocytosis (HLH), and Diamond-Blackfan anemia (DBA). The serum sFasL concentration of age-matched healthy controls was 0.16 +/- 0.11 ng/mL (mean +/- SD, n = 22). The serum sFasL levels in the patients with HLH and DBA were 3.75 +/- 3.82 (n = 19; P < .0001, HLH v control) and 2.76 +/- 2.43 ng/mL (n = 6; P = .012, DBA v control), respectively. Serum interferon-gamma concentration was elevated in the patients with HLH (1.61 +/- 2.62 ng/mL) but not in those with DBA (below the detectable level). These results suggest that the Fas-FasL system plays a role, at least in part, in the pathophysiology of HLH and DBA.

Ultrastructural and Cytochemical Characteristics of Phagocytes in Kuruma Prawn.

Five types of phagocytes were observed in kuruma prawn Penaeus japonicus. Two of them were fixed phagocytes and three were hemocytes. Fixed phagocytes were detected in the heart and lymphoid organ. Phagocytes in the heart were observed on basal lamina covering sarcolemma of heart muscles and the cells had many lysosomal granules (0.1 μm in diameter). Phagocytes in the lymphoid organ was characterizedby numerous interdigitating cell processes and few cytoplasmic granules. The hemocytes were classified into three types, hyaline cell (HC), small granular cell (SGC) and large granular cell (LGC), according to the presence of cytoplasmic deposit and size of granules. The HC had cytoplasmic deposits and a few small granules (0.2 μm in diameter). The SGC had small granules (0.2 μm in diameter) but the LGC contained large granules (1 μm in diameter). Lysosomal enzymes, such as acid phosphatase, β-glucuronidase and α-naphthyl butylate esterase, were detected in all five phagocytic cell types. Esterase activity was higher in the LGC than in any other phagocytes. Peroxidase activity was also detected in all phagocytes, although it was weak. Prophenoloxidase activity was found only in the SGC and LGC. These results indicate that the fixed phagocytes are different from the hemocytes both in their ultrastructural and cytochemical characteristics.

Indications for a distinct putative T cell population in mucosal tissue of carp ( Cyprinus carpio L.)

A monoclonal antibody against carp intestinal T cells (WCL38; of IgM class) was produced by immunization of mice with isolated membrane molecules of carp intestinal intraepithelial lymphoid cells. Flow cytometric analysis showed that WCL38 reacted with 50–70% of the lymphoid cells isolated from intestine, gills or skin, with less than 6% of lymphoid cells isolated from thymus, head kidney or spleen and with a negligible number of PBL. WCL38 + cells were abundant in the intestinal epithelium and less numerous in the lamina propria. Immunogold labelling confirmed that WCL38 reacted with lymphoid cells; in gills and skin some of them have the morphology of large granular lymphoid cells. Immunochemical analysis showed that WCL38 reacted with dimeric membrane molecule on mucosal lymphoid cells with an Mr of 76 kDa, consisting of two 38 kDa subunits. WCL 38 + lymphoid cells are postulated to T cells, since WCL38 does not react with B cells, macrophages or non-specific cytotoxic cells. In conclusion, like higher vertebrates, carp seem to have a distinct (putative) T cell population in their mucosal tissues.

Cytokine cascades in acute graft-versus-host disease.

Cytokine cascades in acute graft-versus-host disease.

Cytokine Expression and Tumorigenicity of Large Granular Lymphocytic Leukemia Cells From Mice Transgenic for the tax Gene of Human T-Cell Leukemia Virus Type I

AbstractThe human T-cell leukemia virus type I (HTLV-I) regulatory protein, Tax, has been speculated to play a major role in HTLV-I leukemogenesis. Indeed, several studies have suggested that upregulation of various cellular oncogenes and cytokines by Tax may explain the pathogenesis observed in HTLV-I–infected individuals, as well as several Tax-transgenic animal models. We report here the analysis of cytokine expression in a Tax-transgenic animal model with large granular lymphocytic (LGL) leukemia. Two different transgenic mice showed identical expression of interleukin-1α (IL-1α), IL-1β, interferon γ (IFNγ), and granulocyte-macrophage colony-stimulating factor (GM-CSF ) in peripheral tail tumors. Interestingly, LGL cell lines derived from these same tumors expressed high levels of both IFNγ and GM-CSF, which correlated with the level of Tax expression. These same LGL cell lines also expressed high levels of lymphocyte function-associated antigen-1 (LFA-1) and intracellular adhesion molecule-1 (ICAM-1). Engraftment of these LGL cell lines into severe combined immunodeficient (SCID) mice led to the development of leukemia and lymphomas. Examination of these SCID mice showed that their pathology was nearly identical to that observed in the original Tax-transgenic mouse model. Both the Tax-transgenic and engrafted SCID mouse models allow for the analysis of cellular events that are required for tumor development associated with HTLV infection and suggest that Tax expression may be responsible for the upregulation of certain cytokines and adhesion molecules that affect the infiltrating capabilities of HTLV-I–infected cells.

Cytokine Expression and Tumorigenicity of Large Granular Lymphocytic Leukemia Cells From Mice Transgenic for the tax Gene of Human T-Cell Leukemia Virus Type I

The human T-cell leukemia virus type I (HTLV-I) regulatory protein, Tax, has been speculated to play a major role in HTLV-I leukemogenesis. Indeed, several studies have suggested that upregulation of various cellular oncogenes and cytokines by Tax may explain the pathogenesis observed in HTLV-I–infected individuals, as well as several Tax-transgenic animal models. We report here the analysis of cytokine expression in a Tax-transgenic animal model with large granular lymphocytic (LGL) leukemia. Two different transgenic mice showed identical expression of interleukin-1α (IL-1α), IL-1β, interferon γ (IFNγ), and granulocyte-macrophage colony-stimulating factor (GM-CSF ) in peripheral tail tumors. Interestingly, LGL cell lines derived from these same tumors expressed high levels of both IFNγ and GM-CSF, which correlated with the level of Tax expression. These same LGL cell lines also expressed high levels of lymphocyte function-associated antigen-1 (LFA-1) and intracellular adhesion molecule-1 (ICAM-1). Engraftment of these LGL cell lines into severe combined immunodeficient (SCID) mice led to the development of leukemia and lymphomas. Examination of these SCID mice showed that their pathology was nearly identical to that observed in the original Tax-transgenic mouse model. Both the Tax-transgenic and engrafted SCID mouse models allow for the analysis of cellular events that are required for tumor development associated with HTLV infection and suggest that Tax expression may be responsible for the upregulation of certain cytokines and adhesion molecules that affect the infiltrating capabilities of HTLV-I–infected cells.

Haematological and phenoloxidase activity changes in the shrimpPenaeus stylirostrisin relation with the moult cycle: protection against vibriosis

This paper describes the changes in the total and differential haemocyte counts during the moult cycle of the shrimp,Penaeus stylirostris. Correlative measures of phenoloxidase (PO) activity and resistance to infection against vibriosis were also investigated during the moult cycle. Three types of cells are identified: hyaline cells (HC), which are small with a big nucleus, represent 80% of the total; semi-granular cells (SGC) represent 10–13%; and large granular cells (LGC) represent 4–10% of the total cells, being the largest cells with highly eosinophilic intracytoplasmic granules. Significant variations of total haemocyte count (THC) between intermoult and premoult were recorded; a minimum in the former and a maximum in the latter. The PO activity measured at the same time decreased during the moult cycle but when compared to the THC was higher at intermoult. The variations in PO activity were similar to those of the relative percentage of the LGC, showing its role in the prophenoloxidase system (proPO-system). The resistance to infection was studied by performing anin vitroassay. Experimental infection with a highly pathogenicVibrioAM23 was carried out by bathing selected shrimps in a concentration of 105cfu ml−1at intermoult and premoult. The mortality of shrimps in premoult was 48% while in intermoult the mortality was 21%. This indicated that shrimps infected in premoult were more sensitive to vibriosis than in intermoult. Further, the variations of selected parameters (i.e. THC, PO) seem to be consistent with thein vivoresponse against the highly pathogenicVibrioAM23.

Autoantibody Production in Healthy Elderly People is not Promoted by Interleukin‐10 Although This Cytokine is Expressed in Them by a Peculiar CD8+CD3+ Large Granular Cell Subpopulation

Healthy elderly people tend to have autoantibodies in their sera. These antibodies, not being associated with any clinical manifestation, have been considered as natural autoantibodies. In systemic lupus erythematosus, as well as in rheumatoid arthritis, the presence of autoantibodies characteristic of these disease (anti-dsDNA and rheumatoid factor, respectively) depends on the endogeneous production of IL-10. The same could hold true for autoantibodies found in healthy elderly individuals. In the present work, the authors analysed whether an increased production of IL-10 contributed to the production of autoantibodies in elderly people. The authors found that there is neither increased in vivo gene expression nor augmented production of IL-10 by peripheral blood mononuclear cells from elderly women even if they do produce autoantibodies. The authors further sought to determine if the production of autoantibodies is inhibited in vitro by adding an anti-IL-10 MoAb to cell cultures and found that it is not. Despite these negative findings of a role for IL-10 in the production of autoantibodies in elderly people, the authors investigated which cells produce IL-10. In so doing they found that intracellular IL-10 expression occurred exclusively in monocytes in young female controls, but in elderly females it involved also CD8+CD3+ large granular cells. These results indicate that autoantibody production in healthy aged individuals is IL-10 independent.

Retroperitoneal granular cell tumor computed tomography findings with 5-year follow-up

We describe the computed tomography (CT) appearance of an unusual, large retroperitoneal granular cell tumor and provide a 5-year follow-up. The tumor occurred in association with multiple, smaller, subcutaneous granular cell tumors. The CT findings were nonspecific, but the scans delineated the extent of the lesion and its relationship to adjacent vessels well. To our knowledge, this report is the first description of the CT appearance of retroperitoneal granular cell tumor in the medical literature.

Human thymocyte responsiveness to stem cell factor: synergy with interleukin-2 for the generation of NK/LAK cytotoxicity

Human recombinant stem cell factor (SCF) increases the viability and cell size of a subset of thymocytes in vitro, but does not independently induce phenotypic changes on thymocytes indicative of T cell differentiation. The SCF-responsive thymocytes have characteristics of large granular cells, that do not express T, B or NK cell-related antigens, and are primarily found in immature thymocyte subsets. These large granular thymocytes do not display cytotoxic activity. However, SCF acts synergistically with IL-2 in the generation of cytotoxic effector cells from thymocyte precursors. Synergy in cytotoxicity is observed to both NK-sensitive and NK-resistant targets. Studies of the SCF receptors on thymocytes show that receptors are expressed on mature ‘bright’ CD3 + cells, immature ‘dim’ CD3 + cells as well as CD3 − cells. IL-2 increases the frequency of SCF receptor-positive cells in cultured thymocytes, which may explain its synergy with SCF in the generation of NK/LAK cytotoxicity. These data show that SCF enhances the functional development of thymic NK/LAK cells in vitro.

Failure to reject an allografted tumor after elimination of macrophages in mice.

After an i.p. transplantation of an allogeneic tumor (Meth A) to C57BL/6 mice, a macrophage (M phi)-rich, non-T, non-NK cell population is induced as the major infiltrate and cytotoxic cells. We here evaluated the role of the M phi s in the rejection of allografted Meth A cells and characterized the M phi s in comparison with other well-known M phi s. At all time intervals after transplantation, the highest cytotoxic activities against Meth A tumor were obtained with the M phi-rich population. In addition, the lymphocyte-rich population had a significant but low cytotoxic activity, whereas two other population types, granulocytes and large granular cells, were inactive. When the M phi-rich or the T cell-depleted M phi-rich population was i.p. transplanted simultaneously with Meth A cells into untreated C57BL/6 mice, the tumor cells were rejected without growth. After specific elimination of M phi s by in vivo application of dichloromethylene diphosphonate-containing liposomes, the cytotoxic activity against Meth A cells was hardly induced at the transplantation site of Meth A cells and the allografted Meth A tumor continued to grow, indicating that a type of M phi is the effector cell essential for the rejection. In contrast to other well-known M phi s, the cytotoxic activity against Meth A cells was cell-to-cell contact dependent and soluble factor (e.g., NO and TNF-alpha) independent. Moreover, the cytotoxic activity of the M phi s (H-2b) against 51Cr-labeled Meth A (H-2d) cells was inhibited by the addition of unlabeled H-2d, but not H-2b, H-2k or H-2h, lymphoblasts as well as Meth A cells, implying the specific interaction of the M phi s with H-2d cells.

Human Dendritic Cell Differentiation Pathway From CD34+ Hematopoietic Precursor Cells

The most effective antigen-presenting cells for T lymphocytes are dendritic cells (DCs), the differentiation pathway of which, however, is incompletely characterized. We examined here how DCs differentiated from human cord blood CD34+ progenitor cells cultured with granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha, and stem cell factor. After 5 days, 2 of 3 nonadherent cells were CD13hiHLA-DRhiCD4+, half of them were also CD14+, and < or = 10% were CD1a+. When day-5 sorted CD13hiCD1a- and CD13lo cells were further cultured, CD1a+ cells appeared in the already CD13hi population, whereas CD13hi cells, a minority of which rapidly became CD1a+, emerged from the CD13lo population. By day 12, still 66% of bulk cells in suspension were CD13hi, most of which displayed high forward and side scatters of large granular cells. Half of CD13hi cells were CD1a+. All CD13hi cells expressed to the same extent DR, CD4, costimulatory and adhesion molecules, and various amounts of CD14. CD1a+ cells stimulated allogeneic lymphocytes more than CD13hiCD1a- cells and, although they were CD14+, both cell types were nonspecific esterase-negative nonphagocytic cells and were stronger mixed leukocyte reaction stimulators than were their macrophage counterparts. Eventually, the percentage of CD1a+ cells decreased. However, typical CD1a+ DCs still emerged in culture of sorted day-12 CD13hiCD1a- cells, and adding interleukin-4 to bulk cultures at that time led to the persistence of the CD1a+ population while diminishing CD14 expression. Thus, this system results first in the differentiation of CD13hi precursors that strongly express DR and CD4, from which more mature CD1a+ DCs continuously differentiate all along the culture period.

The novel immunomodulator, Linomide, stimulates interleukin-2-induced human natural killer (NK) cell and PHA-stimulated T cell proliferation from normal donors

Donor-derived cell-mediated immunotherapy has been shown to be an effective tool for reinduction of remission in chronic myeloid leukemia (CML) patients who have relapsed post-bone marrow transplantation (BMT). Linomide, quinoline-3-carboxamine (LS 2616), is a new immunomodulator shown to increase the number of NK precursors in mice in addition to upregulating the quantity of CD56 +, CD3 − and CD16 + NK cells in the peripheral blood of patients following autologous BMT (ABMT). We investigated the in vitro effects of Linomide on NK activity of normal human donors. Large granular lymphocytes (LGLs) and NK cells were incubated overnight with Linomide (0.02–4.8 mg/ml), recombinant human interleukin-2 (IL-2, 75 IU/ml), or a combination of both. Linomide, at 0.02–0.3 mg/ml, augmented IL-2-induced proliferation of LGLs and NK cells in an inversely proportional manner. In contrast, Linomide at 0.6–4.8 mg/ml inhibited IL-2-induced proliferation of LGLs and NK cells in a dose-dependent manner. Linomide was able to potentiate phytohemaglutinin-induced CD3 + T cell proliferation. In addition, supernatants derived from Linomide treated CD3 + T cells were able to mimic the direct stimulatory effect of Linomide on activated NK cell proliferation. These supernatants were found to have low levels of tissue necrosis factor-alpha (TNF-α) and interleukin-1 (IL-1) and therefore Linomide stimulation of NK and T cell proliferation may be due to its inhibitory effect on the secretion of these cytokines by activated CD3 + T cells. Linomide had no effect on cytotoxicity nor on the phenotypic expression of resting and IL-2-activated LGLs or NK cells. In view of our results, Linomide could possibly play a potential role in adoptive cell-mediated immunotherapy post-BMT.

Familial melanoma in the progeny of a Duroc and Slovak white crossbred sow

In three successive litters of a Duroc and Slovak White crossbred sow there was a 7.75% overall incidence of melanoma. Immunohistochemical labelling confirmed the mesenchymal origin of the neoplastic cells, indicated the absence of CD3 lymphocytes in the tumour, and proved the tumorous origin of surrounding non-pigmented cells. An abundance of macrophages (epithelioid cells) with increased endoplasmic reticulum was observed by electron microscopy, but large granular lymphocyte-like cells were few in number.

Isolation and culture of heart cells from the European flat oyster,Ostrea edulis

The present study reports a culture technique for heart tissues of the European flat oyster,Ostrea edulis. Heart tissues of flat oysters were dissociated by a trypsin-EDTA treatment and a mechanical action in a Dounce-type homogeneizer. Then, dissociated cells were cultured in three different synthetic media, in pure sea water or in sea water mixed with sterile filtered Japanese oyster,Crassostrea gigas, hemolymph. All these media were supplemented with 10% of fetal bovine serum. Cultures were grown at 20°C in previously Poly-D-Lysin coated flasks or culture wells. The optimized medium, 3% L15 medium (w/v) in sterile sea water mixed with Japanese oyster hemolymph (1:1) and supplemented with 10% of fetal bovine serum gave the best result. Morphological characterization for the cardiac cultured cells was performed. Thus, cell monolayers of dissociated heart tissues consisted essentially of hemocytes and large granular pigmented cells.