4.Flow cytometric immunophenotyping in fine-needle aspiration of lymph nodes.
Jae Gul CHUNG ; Gyung Yub GONG ; Joo Ryung HUH ; Shin Kwang KHANG ; Jae Y RO
Journal of Korean Medical Science 1999;14(4):393-400
Fine-needle aspiration (FNA) of lymph nodes has been regarded as a useful method in the diagnosis of lymphadenopathy. However, this procedure has been shown to be of limited value in the diagnosis of low or intermediate grade malignant lymphomas in some studies. Immunophenotyping is an essential adjunct to cytomorphology for the diagnosis of lymphoma by FNA. Immunophenotyping using flow cytometry (FCM) is rapid, objective and reliable. Using FCM, multiparametric analysis of 33 FNA materials from lymph nodes was performed and profiles of surface markers of lymphoid cells were assessed. In reactive hyperplasia, patterns of cell surface markers were quite variable, but disclosed polyclonality. Most of the B-cell lymphomas showed immunophenotypes for B-cell lineages with their kappa: lambda or lambda: kappa ratio being over 3:1. In T-cell lymphomas, T-cell surface markers were predominantly expressed as well. In conclusion, our results suggest that immunophenotyping of lymph node aspirates is a valuable diagnostic adjunct for lymphoproliferative disorders, particularly in B-cell lymphomas because immunophenotyping can be easily and adequately performed by FCM.
Antigens, CD19/analysis
;
Antigens, CD20/analysis
;
Antigens, CD3/analysis
;
Antigens, CD4/analysis
;
Antigens, CD5/analysis
;
Antigens, CD7/analysis
;
Antigens, CD8/analysis
;
B-Lymphocytes/immunology
;
B-Lymphocytes/chemistry
;
Biopsy, Needle
;
Flow Cytometry/methods*
;
Hodgkin Disease/pathology
;
Human
;
Immunophenotyping
;
Lymph Nodes/pathology
;
Lymph Nodes/chemistry
;
Lymphatic Diseases/pathology*
;
Lymphatic Metastasis/pathology
;
Lymphoma, B-Cell/pathology*
;
Lymphoma, Non-Hodgkin/pathology
;
T-Lymphocytes/immunology
;
T-Lymphocytes/chemistryt
5.Expressions of p53 and p21 in Primary Gastric Lymphomas.
Journal of Korean Medical Science 2001;16(6):731-735
The p21 overexpression is thought to be a consequence of the p53 induced activation of the p21 gene. The immunohistochemical evaluation of p53 and p21 can be a valuable means of assessing the functional status of the p53 gene product. We examined the overexpression of p21 and p53 proteins in primary gastric lymphomas and the correlation with prognosis. A total of 32 cases of gastric lymphomas was classified into low-grade lymphomas of mucosa-associated lymphoid tissue type (n=16) and high-grade B-cell lymphomas (n=16). In low-grade lymphomas, only one case showed p53 positivity and all cases were p21-negative. In high-grade lymphomas, seven cases were p53+/p21- (44%), one case was p53+/p21+ (6%), and eight cases were p53-/p21- (50%). The p53+/p21- cases had a much lower percentage of patients sustaining a continuous complete remission state (3/7, 43%) compared with other cases (6/7, 86%). From these results, we concluded that p21 expression is rare in primary gastric lymphomas. Therefore, p53-positive lymphomas can be assumed as having p53 mutation. And combined studies of p53 and p21 may be used as a prognostic indicator in primary gastric high-grade lymphomas.
Adult
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Aged
;
Antibodies, Monoclonal
;
Female
;
Human
;
Immunohistochemistry
;
Lymphoma, B-Cell/*chemistry/pathology
;
Male
;
Middle Age
;
Peyer's Patches/chemistry/pathology
;
Prognosis
;
Protein p53/*analysis/immunology
;
Proto-Oncogene Protein p21(ras)/*analysis/immunology
;
Stomach Neoplasms/*chemistry/pathology
;
Support, Non-U.S. Gov't
6.Pathologic diagnosis and subtyping of lymphoma in bone marrow biopsies using histologic examination, immunohistochemistry and gene rearrangement studies.
Jia-cheng XIAO ; Xiao-long JIN ; Fei YUAN
Chinese Journal of Pathology 2004;33(2):120-124
<b>OBJECTIVEb>To assess the value of histologic examination, immunohistochemistry and gene rearrangement studies in the diagnosis and subtyping of lymphoma with bone marrow involvement (BMI).
<b>METHODSb>Sixty-two formalin fixed, paraffin embedded bone marrow biopsy specimens were studied. Immunohistochemical and immunoglobulin heavy chain (IgH) and T-cell receptor gene rearrangement studies were performed in each case.
<b>RESULTSb>Chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) demonstrated mainly and interstitial infiltration by dysplastic lymphocytes, with intertrabecular nodular arrangement or in dispersion. Sometimes, pseudofollicles may be noted. A predominantly para- or intertrabecular infiltration by nodules of lymphoma cells was characteristic of follicle center cell lymphoma (FCL) cases. In most lymphoplasmacytoid lymphoma (LPL) cases, there was infiltration by small lymphocytes and plasma cells between bony trabeculae. In marginal zone cell lymphoma (MZL), vague inter- or para-trabecular nodules of polymorphic lymphoma cells with clear cytoplasm might be noted. Small to medium-sized dysplastic lymphocytes, with absence of paraimmunoblasts or pseudofollicles, were the most frequent findings in mantle cell lymphoma (MCL). Hairy cell leukemia (HCL) might be identified by the presence of distinct cell membrane and abundant clear cytoplasm, resulting in a "fried-egg" appearance. Tumor cells with large nuclei and eosinophilic nucleoli were characteristically seen in lymphomatosis diffusa (Hodgkin's disease, HD). In T-cell non-Hodgkin lymphoma with BMI, dispersed or clusters of intertrabecular neoplastic lymphoid cells with clear cytoplasm and gyriform nuclei were often observed. In diffuse large B-cell lymphoma (DLBL), the tumor cells were large and isolated or arranged in diffuse pattern. Immunohistochemically, a panel of markers, including CD3 CD20, and CD79 are valuable for the differential diagnosis of T- and B-cell lymphomas. The neoplastic cells in MCL were cyclin D1- and CD5-positive, while BCL2- and CD10-positivity was characteristic for FCL. CLL/SLL cells might be stained with CD5 and CD23, in addition to CD20 and CD79. CD25 expression might be noted in HCL: the positivity for CD15, CD30 and fascin suggests HD. There was a higher positivity rate for IgH gene rearrangement in CLL/SLL, LPL MZL and DLBL (80%, 60%, 66.7%, 70% respectively) and for T- cell receptor gamma gene rearrangement in T-cell lymphoma (66.7%).
<b>CONCLUSIONb>A combination of histopathology, immunohistochemistry and IgH / T-cell receptor gamma gene rearrangement studies may be of aid to the diagnosis and subtyping of lymphoma with BMI, especially if there is only a small number of tumor cells present in the specimen.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow ; chemistry ; pathology ; Diagnosis, Differential ; Female ; Gene Rearrangement ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Leukemia, Lymphocytic, Chronic, B-Cell ; pathology ; Lymphoma ; classification ; immunology ; pathology ; Lymphoma, Follicular ; pathology ; Male ; Middle Aged
7.Identification of specific peptide ligands for B-lymphoma cell and its effect on tyrosine phosphorylation and cell apoptosis.
Liangwen SONG ; Xianmei MA ; Xuemei CUI ; Yang LI ; Xiaomin WANG
Chinese Medical Sciences Journal 2004;19(1):44-50
<b>OBJECTIVEb>To search novel method for diagnosis and therapy of B-lymphoma, specific small molecular peptide ligands against binding site of tumor cells were screened and its effects on signal transduction and cell apoptosis were tested.
<b>METHODSb>Specific peptide ligands were screened by binding with site of human B lymphoma cell (OC1LY8) using peptide-bead libraries. The identified peptides were characterized with responsible cells by rebinding test. The role of tyrosine phosphorylation of peptide ligand was tested by Western blot; and its apoptosispromoting role was observed by confocal fluorescent microscope.
<b>RESULTSb>Specific peptide ligand was able to bind specifically to site on cell surface and enter into cytoplasm. Tetrameric peptide ligand was able to strongly trigger signal transduction resulting in tyrosine phosphorylation and cellular apoptosis in OC1LY8 cell line.
<b>CONCLUSIONb>Screened peptide ligand can effectively bind with OC1LY8 cell, stimulate cellular tyrosine phosphorylation and induce cellular apoptosis.
Amino Acid Sequence ; Apoptosis ; drug effects ; Cell Line, Tumor ; Humans ; Ligands ; Lymphoma, B-Cell ; metabolism ; pathology ; Oligopeptides ; chemistry ; pharmacology ; Peptide Library ; Phosphorylation ; drug effects ; Signal Transduction ; drug effects ; Tyrosine ; metabolism
8.Expression and cloning of mCD99L2 gene from mouse B lymphoma cell line A20 and construction of its eukaryotic expression vector.
Li-jia SHEN ; Wei-yi FANG ; Si-ming XIE ; Ying HE ; Hui-yong JIANG ; Tong ZHAO
Journal of Southern Medical University 2006;26(2):144-149
<b>OBJECTIVEb>To detect and clone mCD99L2 gene from mouse B lymphoma cell line A20 and construct its eukaryotic expression vector pcDNA3.1-mCD99L2.
<b>METHODSb>The expression of mCD99L2 mRNA in A20 cell line was detected by in situ hybridization. The total RNA of A20 cells was extracted to obtain the full-length cDNA of the coding region of mCD99L2 gene by RT-PCR, the product of which was ligated into pMD18-T vector and the DNA sequence of the insert was detected. The coding regions of mCD99L2 gene was amplified from pMD-mCD99L2 by PCR using primers containing EcoR I and Xho I sites and cloned into the eukaryotic expression vector pcDNA3.1/MycHis(+).
<b>RESULTSb>In situ hybridization identified positive expression of mCD99L2 gene in the A20 cell line. The full-length cDNA of mCD99L2 coding region of A20 cell line was obtained by RT-PCR, which yielded a product of 712 bp as expected, and the DNA sequence was completely homologus to the mCD99L2 cDNA reported in GenBank. Restriction endonuclease digestion and DNA sequencing indicated that the eukaryotic expression vector pcDNA3.1(+)- mCD99L2 had been constructed successfully.
<b>CONCLUSIONb>mCD99L2 cDNA has been cloned from mouse B lymphoma cell line A20 and its eukaryotic expression vector pcDNA3.1(+)- mCD99L2 successfully constructed, which facilitates further functional study of mCD99L2 gene in mouse B lymphoma cell line A20.
12E7 Antigen ; Animals ; Antigens, CD ; biosynthesis ; genetics ; Base Sequence ; Cell Line, Tumor ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Eukaryotic Cells ; metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; genetics ; Lymphoma, B-Cell ; genetics ; pathology ; Mice ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA
9.Synthesis and antiviral activities of geldanamycin analog TC-GM in vitro.
Chun-Xin LI ; Guang-Zhi SHAN ; Bo FAN ; Pei-Zhen TAO ; Li-Xun ZHAO ; Jian-Dong JIANG ; Yu-Huan LI ; Zhuo-Rong LI
Acta Pharmaceutica Sinica 2011;46(6):683-687
In order to find antiviral compounds with novel structures, geldanamycin and lamivudine with different antiviral mechanisms were conjunctively synthesized to acquire a new compound TC-GM, and the antiviral activity of TC-GM was measured. The antiviral activity against HIV-1 was examined by p24 antigen ELISA kit. The activity against HBV was examined by dotblot. The activity against HSV and CoxB virus was examined by CPE. TC-GM exhibited broad-spectrum antiviral activities similarly like geldanamycin. TC-GM inhibited the replication of different viruses, including HIV-1, HBV, HSV 1 and 2, CoxB6. TC-GM showed more potent inhibitory activity against HIV-1 and HBV than other detected virus.
Animals
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Anti-HIV Agents
;
chemical synthesis
;
chemistry
;
pharmacology
;
Antiviral Agents
;
chemical synthesis
;
chemistry
;
pharmacology
;
Benzoquinones
;
chemical synthesis
;
chemistry
;
pharmacology
;
Cell Line, Tumor
;
Cercopithecus aethiops
;
Enterovirus B, Human
;
drug effects
;
physiology
;
HIV-1
;
drug effects
;
physiology
;
Hep G2 Cells
;
Hepatitis B virus
;
drug effects
;
physiology
;
Herpesvirus 1, Human
;
drug effects
;
physiology
;
Herpesvirus 2, Human
;
drug effects
;
physiology
;
Humans
;
Lactams, Macrocyclic
;
chemical synthesis
;
chemistry
;
pharmacology
;
Lamivudine
;
chemical synthesis
;
chemistry
;
pharmacology
;
Madin Darby Canine Kidney Cells
;
Precursor Cell Lymphoblastic Leukemia-Lymphoma
;
pathology
;
virology
;
Vero Cells
;
Virus Replication
;
drug effects
10.Clinical significance of nuclear factor kappaB and chemokine receptor CXCR4 expression in patients with diffuse large B-cell lymphoma who received rituximab-based therapy.
Ho Cheol SHIN ; Jongwon SEO ; Byung Woog KANG ; Joon Ho MOON ; Yee Soo CHAE ; Soo Jung LEE ; Yoo Jin LEE ; Seoae HAN ; Sang Kyung SEO ; Jong Gwang KIM ; Sang Kyun SOHN ; Tae In PARK
The Korean Journal of Internal Medicine 2014;29(6):785-792
BACKGROUND/AIMS: This study investigated the expression of nuclear factor kappaB (NF-kappaB) and the chemokine receptor (CXCR4) in patients with diffuse large B-cell lymphoma (DLBCL) who received rituximab-based therapy. METHODS: Seventy patients with DLBCL and treated with rituximab-CHOP (R-CHOP) were included, and immunohistochemistry was performed to determine the expression of NF-kappaB (IkappaB kinase alpha, p50, and p100/p52) and CXCR4. To classify DLBCL cases as germinal center B-cell-like (GCB) and non-GCB, additional immunohistochemical expression of CD10, bcl-6, or MUM1 was used in this study. The expression was divided into two groups according to the intensity score (negative, 0 or 1+; positive, 2+ or 3+). RESULTS: The median age of the patients was 66 years (range, 17 to 87), and 58.6% were male. Twenty-seven patients (38.6%) had stage III or IV disease at diagnosis. Twenty-three patients (32.9%) were categorized as high or high-intermediate risk according to their International Prognostic Indexs (IPIs). The overall incidence of bone marrow involvement was 5.7%. Rates of positive NF-kappaB and CXCR4 expression were 84.2% and 88.6%, respectively. High NF-kappaB expression was associated with CXCR4 expression (p = 0.002), and 56 patients (80.0%) showed coexpression. However, the expression of NF-kappaB or CXCR4 was not associated with overall survival and EFS. On multivariate analysis that included age, gender, performance status, stage, and the IPI, no significant association between the grade of NF-kappaB or CXCR4 expression and survival was observed. CONCLUSIONS: The current study suggests that the tissue expression of NF-kappaB and CXCR4 may not be an independent prognostic marker in DLBCL patients treated with R-CHOP.
Adolescent
;
Adult
;
Aged
;
Aged, 80 and over
;
Antibodies, Monoclonal, Murine-Derived/administration & dosage
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Antineoplastic Combined Chemotherapy Protocols/adverse effects/*therapeutic use
;
Chi-Square Distribution
;
Cyclophosphamide/administration & dosage
;
Disease Progression
;
Disease-Free Survival
;
Doxorubicin/administration & dosage
;
Female
;
Humans
;
Immunohistochemistry
;
Kaplan-Meier Estimate
;
Lymphoma, Large B-Cell, Diffuse/chemistry/*drug therapy/mortality/pathology
;
Male
;
Middle Aged
;
Multivariate Analysis
;
NF-kappa B/*analysis
;
Neoplasm Staging
;
Predictive Value of Tests
;
Prednisone/administration & dosage
;
Proportional Hazards Models
;
Receptors, CXCR4/*analysis
;
Retrospective Studies
;
Risk Factors
;
Time Factors
;
Treatment Outcome
;
Tumor Markers, Biological/*analysis
;
Vincristine/administration & dosage
;
Young Adult