OBJECTIVETo search a method for showing dust granules in phagocytes.

METHODSThe autopsy samples of 10 cases who died of non-pulmonary diseases(group A), 6 cases with silicosis(group B) and 5 cases of dead fetus(group C) were collected. The samples of lung, spleen and lymphnodes were stained by methods of (1)HE staining; (2)Warthin-Starry staining(W-S staining) and modified W-S staining; (3) CD68 (as a first antibody) immunohistochemical staining. The ultrastructure and chemical component of the dust granules in the pulmonary phagocytes(dust cells) of group A(3 cases) and group B(3 cases) were observed and analyzed by transmission electron microscope(TEM) and analytic electron microscope(AEM).

RESULTSIn group A and B smaller or minute dust granules which could not be shown by HE staining method were clearly shown by W-S staining and modified W-S staining methods. There were no positive granules found in group C. The CD68 marker of the dust cells was positive. The size, shape, density and chemical elements of the dust granules of group A were different from those of group B under TEM and AEM, excluding bacteria and other intracellular contents.

CONCLUSIONThe dust granules in the phagocytes could be shown by W-S staining method, its staining effect for minute dust granules is superior to general HE staining. However, the determination of chemical elements of dusts must depend on AEM.

Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Dust ; Humans ; Microscopy, Electron ; Phagocytes ; ultrastructure ; Silicosis ; pathology ; Staining and Labeling ; methods

OBJECTIVETo explore the clinicopathologic features, immunophenotype, differential diagnosis and gene mutation status of the Erdheim-Chester disease (ECD).

METHODSClinical and pathologic findings of 3 ECD cases were examined by gross, microscopic, immunohistochemical methods and BRAF V600E mutation. Related literatures were reviewed.

RESULTSTwo male patients and one female patient presented clinically with multiple skin nodules, bone pain and bony lesions by imaging study. Microscopically, the lesions were composed of spindle-shaped fibroblasts, foamy histiocytes and scattered Touton-type giant cells embedded in reactive fibrous tissue. Lymphocytes, plasma cells, and multinucleated giant cells were also found. Immunohistochemically, all histiocytes were positive for CD68, none of which expressed CD1a, although 2 cases focally expressed weak S-100 stain. In 2 cases,BRAF V600E mutation was detected.

CONCLUSIONSECD is a rare disease of xanthogranulomatous histiocytosis.Its diagnosis relies on pathological and immunohistochemical findings, but correlation with clinical information, especially radiographic findings should be performed.No effective treatment of the disease is currently available.

Antigens, CD ; analysis ; Antigens, CD1 ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Diagnosis, Differential ; Erdheim-Chester Disease ; genetics ; immunology ; pathology ; Female ; Humans ; Male ; Mutation ; S100 Proteins ; analysis ; Treatment Outcome

Pterygium is a proliferative disease. Recent research has reported that stem cells are involved in the pathogenesis of various proliferative diseases, including solid tumors and diabetic proliferate vitreoretinopathy. In previous literature, we hypothesized that adult stem cells originated from bone marrow were involved in the pathogenesis of pterygium. We proved this by immunohistochemical staining with various stem cell markers. The staining showed adult stem cells in the pterygium. c-kit positive cells were observed primarily in the stroma, and some cells were also found in the basal epithelium. AC133 and CD34 positive cells were primarily found in the basal epithelium and were ovoid shaped, similar to the c-kit cells. However, some cells were found in vascular endothelium. STRO-1 positive cells were found mainly in the stroma and were spindle shaped. In recurrent pterygium, cells were more scattered and the expression pattern was denser. Therefore, we suggest a new theory of pterygium pathogenesis. Inflammation caused by environmental factors triggers the abnormal production of some growth factors and cytokines in order to recover from cellular damage. If these healing signals are excessive, limbal basal cells will be changed to abnormally-altered pterygial cells. The excessive wound healing process and remnant altered cells result in recurrence using the same mechanism.
Stem Cells/*physiology ; Pterygium/*etiology ; Proto-Oncogene Proteins c-kit/analysis ; Peptides/analysis ; Middle Aged ; Humans ; Glycoproteins/analysis ; Bone Marrow Cells/*physiology ; Antigens, CD34/analysis ; Antigens, CD/analysis

To characterize cellular responses during hepatic regeneration, we examined 13 explant livers and 5 liver allografts by immunohistochemistry for cytokeratin 7, HepPar1, CD68, alpha-smooth muscle actin (alpha-SMA) and proliferating cell nuclear antigen as well as reticulin and Masson-trichrome staining. Within a week after liver damage, elongated CD68-positive cells were detected along the border of necrotic area. The number of alpha-SMA-positive cells was slightly increased along the sinusoids. Ductular proliferation or fibrosis was negligible. After one or two weeks, the size and number of CD68-positive cells were markedly increased. alpha-SMA-positive cells increased in number within lobules and portal tracts. Ductular proliferation occurred predominantly at the limiting plate or along the border of necrotic areas. After one month, necrotic parenchyma was replaced by many ductules, CD68-positive cells, alpha-SMA-positive cells. Nodules of regenerating hepatocytes and irregular fibrosis were diffusely present. Other nonparenchymal cells were not significantly changed. These observations indicate that chronological interaction between nonparenchymal and parenchymal cells occur during the course of human hepatic regeneration and suggest extensive porto-periportal fibrosis more than a few months after the onset of fulminant hepatitis is a major indicator of chronic functional impairment necessitating liver transplantation.
Actins/analysis ; Antigens, CD/analysis ; Antigens, Differentiation, Myelomonocytic/analysis ; Human ; Immunohistochemistry ; Keratin/analysis ; Liver/*cytology ; *Liver Regeneration ; Proliferating Cell Nuclear Antigen/analysis

Flowcytometer has been flourishing since the passed decade in our nation. Many general hospitals use it for routine tests. One of the main usage is immunophenotyping for hematological malignancies. Flowcytometric immunophenotyping (FCM-IM) facilitates accurate diagnosis in both leukemias and lymphoid proliferative disorders (LPDs). It becomes an indispensable criteria in the diagnosis of hematological malignancies. It may also offer additional therapeutically relevant information. Typical patterns of immunophenotype in leukemias and certain types of LPDs are illustrated in figure 1 and figure 2. But variation or overlapping appears often. In order to reach a definitive diagnosis and to have a common interpretation between laboratory to laboratory the following items need not to be ignored: 1. Direct staining and multi-color or multi-parameter flow cytometric analysis is necessary. 2. The phenotype of abnormal cells should be determined on as pure a population as possible therefore CD45/SSC gating is critical for isolating the blast population (abnormal cells). CD45/SSC gating is much better than FSC/SSC gating especially when percentage of blast is low in the sample. Else, each subtype of leukemia has its special CD45/SSC pattern. 3. The phenotype of this certain population should be described clearly but it is not necessary to count the percentage of each marker. 4. > 20% for positive threshold used in indirect staining method may be confused in multi-color direct staining method. 5. Hybrid leukemia diagnosed by FCM only should be in caution since the divergence of antigen/antibody, the lineage infidelity of antigen expression and the biological characteristics of the leukemic cells. Flowcytometer is not a cell counter. "It provides a pattern of information that must be interpreted by a knowledgeable professional in the context of the clinical situation".
Antigens, CD ; analysis ; Diagnostic Techniques and Procedures ; Flow Cytometry ; Humans ; Immunophenotyping ; Leukemia ; classification ; pathology ; Reference Standards

OBJECTIVESTo evaluate the specificity of three-color flow cytometry in childhood acute lymphoblastic leukemia (ALL) immunophenotyping.

METHODSImmunophenotyping was performed by three-color flow cytometry analysis using CD(45)/SSC gating.

RESULTSThe percentage of blasts was correlated better with leukemic cell count compared with that of FSC/SSC, and the false positive results were low. Among eighty six cases of ALL, 95.3% was B-ALL, in which common-ALL and Pro-B-ALL were 76.8% and 6.1%, respectively, and 2.3% was T-ALL. CD(34)(+) and myeloid-associated antigen expression were observed in 57.0% and 34.9% of the cases, respectively, among which Pro-B-ALL was the commonest. CD(33) was more commonly expressed than CD(13) in Pro-B-ALL cases, but no difference in the expression between these two antigens in other subtypes.

CONCLUSIONGating of CD(45)/SSC eliminated effection of normal cells to blasts in bone marrow, with which the immunophenotyping results were more reliable.

Antigens, CD ; analysis ; Child ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology

OBJECTIVETo define the immune phenotype of colon cancer cells.

METHODSUsing a panel of 40 anti-human monoclonal antibodies (MoAbs), the cells of colon cancer HR8348 were analyzed with three-color flow cytometry after direct immunofluorescent staining.

RESULTSHR8348 cell line did not express CD2, CD3, CD4, CD5, CD7, CD8, TCR, CD10, CD11b, CD14, CD16, CD19, CD22, CD25, CD28, SmIg, CD33, CD35, CD36, CD41a, CD45, CD45RA, CD45RO, CD56, CD61, CD64, CD66b, CD69, CD71, CD117, CD122 and P-glycoprotein but expressed CD13, CD15, CD20, CD38, CD95 and HLA-DR.

CONCLUSIONThe results demonstrate that colon cancer cell line HR8348 shares some antigenic determinants with leucocyte lineage.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Antigens, CD ; analysis ; Cell Line, Tumor ; Colonic Neoplasms ; chemistry ; Humans

Asthma is a chronic disease of airway inflammation due to excessive T helper cell type 2 (Th2) response. Present treatment based on inhalation of synthetic glucocorticoids can only control Th2-driven chronic eosinophilic inflammation, but cannot change the immune tolerance of the body to external allergens. Regulatory T cells (Tregs) are the main negative regulatory cells of the immune response. Tregs play a great role in regulating allergic, autoimmune, graft-versus-host responses, and other immune responses. In this review, we will discuss the classification and biological characteristics, the established immunomodulatory mechanisms, and the characteristics of induced differentiation of Tregs. We will also discuss the progress of Tregs in the field of asthma. We believe that further studies on the regulatory mechanisms of Tregs will provide better treatments and control strategies for asthma.
Antigens, CD/analysis* ; Apyrase/analysis* ; Asthma/immunology* ; Cell Differentiation ; Cytokines/metabolism* ; Humans ; Lymphocyte Transfusion ; T-Lymphocytes, Regulatory/immunology*

OBJECTIVETo study the clinical significance of CD163 in the diagnosis and the evaluation of severity and prognosis of childhood hemophagocytic lymphohistiocytosis (HLH).

METHODSNinety-four children were classified into Epstein-Barr virus (EBV)-positive (n=55) and EBV-negative groups (n=39; control group). The EBV-positive group was subgrouped into infectious mononucleosis (IM; n=47) and HLH (n=8). Serum levels of soluble CD163 were measured using ELISA. Expression of CD163 on mononuclear cells was detected by flow cytometry.

RESULTSThe serum levels of soluble CD163 were>10 000 ng/mL in all eight HLH patients (>30 000 ng/mL in 3 cases). The mean serum levels of soluble CD163 in the HLH group were significantly higher than in the control and IM groups (P<0.05). The serum levels of soluble CD163 in EBV-positive children were positively correlated with EBV-DNA copies and serum levels of ferritin and LDH, but were negatively correlated with white blood cell count, neutrophil count, hemoglobin and platelet count. The follow-up after treatment for three HLH patients showed that serum levels of soluble CD163 were significantly reduced, but the soluble CD163 levels rebounded in one patient who was complicated by fungal pneumonia infection.

CONCLUSIONSThe levels of serum soluble CD163 may be related to the severity in children with HLH. The EBV-positive children with soluble CD163 levels >10 000 ng/mL should be considered the possibility of HLH.

Adolescent ; Antigens, CD ; analysis ; Antigens, Differentiation, Myelomonocytic ; analysis ; Child ; Child, Preschool ; Epstein-Barr Virus Infections ; immunology ; Female ; Humans ; Infant ; Male ; Receptors, Cell Surface ; analysis

This study was to investigate dynamics of biological properties of CD133(+) cells from human umbilical cord blood (UCB) during short-term culture containing the combination of hematopoietic growth factors and the feasibility of in vitro expansion of CD133(+) cells. The biology activities including analysis of cell cycle, immunophenotype, telomerase activity, expression of adhesion molecules and expansion potential of CD133(+) cells were monitored during ex-vivo expansion, and compared with those of CD34(+) cells. The results showed that the contents of CD133(+) and CD34(+) cells in fresh UCB were (1.05 +/- 0.73)% and (1.40 +/- 0.56)% respectively. About 79.62% of CD34(+) cells expressed CD133, and more than 97% of CD133(+) cells were CD133(+)CD34(+), markedly higher than that in CD34(+) fraction (P < 0.01). No significant differences were observed in content of cells expressing CD38, CD13, CD14, CD61 and glycophorin-A between the two fractions. Expansion of CD133(+), CD133(+)CD34(+) and CD34(+)CD38(-) cells at 10 days and those of CFU-mix, HPP-CFC and CD34(+)CD38(-) cells at 6 days from CD133(+) cells group were significantly higher than those from the CD34(+) cell group (P < 0.05). Analysis of immunophenotype showed that CD133(+)CD34(+) cells declined gradually while CD133(-)CD34(+) and CD133(-)CD34(-) cells increased during ex-vivo expansion; basal telomerase activities of fresh UCB CD133(+) and CD34(+) cells were low but significantly exceeded that of CD34(-) fraction (P < 0.05). At first week of expansion, telomerase activity was significantly upregulated, after two weeks, telomerase activity remarkably declined, and decreased to baseline or below the limits of detection in day 20. More than 90% of CD133(+) cells expressed CD49d and CD11a, and, more than 85% of the cells expressed CD54, about 50% of cells expressed CD62L. At the early stage of expansion, expression of CD49d was upregulated, expression of CD11a remaining no change, while as expression of CD54 and CD62L was downregulated. Expression of all adhesion molecules was decreased gradually with extend of culture. But expression of these adhesion molecules on CD34(+) subsets were not affected significantly during expansion. It is concluded that CD133(+) population may be a more primitive hematopoietic stem/progenitor cells (HSPC) than CD34(+) cells, CD133(+) cells have great expansion potential for ex-vivo expansion and is a suitable target cell for ex-vivo expansion of HSPC. Downregulation of adhesion molecules and telomerase activity may be one of the reasons for delayed engraftment of expanded products.
AC133 Antigen ; Antigens, CD ; Antigens, CD34 ; analysis ; Cell Cycle ; Cells, Cultured ; Fetal Blood ; cytology ; Glycoproteins ; analysis ; Hematopoietic Stem Cells ; physiology ; Humans ; Immunophenotyping ; Peptides ; analysis ; Telomerase ; metabolism