Wen-Qing Wang

Parvovirus B19 infection associated with Hashimoto's thyroiditis in adults.

OBJECTIVEnParvovirus B19 is a common human pathogen, which has been linked to autoimmune diseases recently. The aim of the study is to evaluate whether B19 is involved in adult Hashimotos thyroiditis (HT).nnnMETHODSnEighty-six thyroid tissues from the adult patients with a spectrum of thyroid disorders were examined for B19 DNA and capsid protein by nested PCR, in-situ hybridization and immunohistochemistry. The presence of viral DNA in HT epithelium was studied by laser-capture microdissection and sequencing of PCR products. The expressions of nuclear factor-kappaB (NF-kappaB) and interleukin-6 were investigated by immunohistochemistry.nnnRESULTSnB19 DNA was significantly present in HT tissues by both PCR (29/32, 90.6%) and in-situ hybridization (23/32, 71.9%, all p < 0.01) compared with normal thyroid tissue (7/16, 43.8%; 2/16, 12.5%). Laser-capture microdissection further confirmed this difference. B19 capsid protein in HT group was significantly higher than that in all the control groups (p < 0.01), and the expression of NF-kappaB and interleukin-6 in HT tissues was up-regulated. NF-kappaB was well co-localized with B19 protein in thyroid epithelia by double-labeling immunofluorescence and confocal microscopy.nnnCONCLUSIONSnThe presence of B19 nuclear acid and viral protein was significantly common in HT tissues and it suggested a possible role of B19 in adult HT.

Detection of human parvovirus B19 in papillary thyroid carcinoma

To evaluate whether parvovirus B19, a common human pathogen, was also involved in papillary thyroid carcinoma (PTC), 112 paraffin-embedded thyroid specimens of benign nodules, papillary, medullary and follicular carcinomas, and normal controls were examined for B19 DNA and capsid protein by nested PCR, in situ hybridisation (ISH) and immunohistochemistry (IHC). The expression of the nuclear factor-κB (NF-κB) was investigated by IHC. The results showed B19 DNA commonly exists in human thyroid tissues; however, there were significant differences between PTC group and normal controls, and between PTC and nonneoplastic adjacent tissues (P<0.001). The presence of viral DNA in PTC neoplastic epithelium was confirmed by laser-capture microdissection and sequencing of nested PCR products. B19 capsid protein in PTC group was significantly higher than that of all the control groups and nonneoplastic adjacent tissues (P⩽0.001). Compared with control groups, the activation of NF-κB in PTC group was significantly increased (P⩽0.02), except for medullary carcinomas, and the activation of NF-κB was correlated with the viral protein presence (P=0.002). Moreover, NF-κB was colocalised with B19 DNA in the neoplastic epithelium of PTC by double staining of IHC and ISH. These results indicate for the first time a possible role of B19 in pathogenesis of PTC.

Detection of parvovirus B19 nucleic acids and expression of viral VP1/VP2 antigen in human colon carcinoma.

OBJECTIVES:The aim of this study was to evaluate whether parvovirus B19, a common infectious pathogen in humans, also was involved in human colon carcinoma.METHODS:A total of 119 paraffin-embedded specimens of colon polyps, adenocarcinomas, carcinoma-adjacent tissues, and normal controls were processed for nested polymerase chain reaction (PCR), in situ hybridization (ISH), immunohistochemistry (IHC), and laser capture micro dissection detection of B19 DNA and protein. The expression of cyclo-oxygenase-2 (COX-2) in the colon- cancer cells (Lovo) transfected by inducible vector for VP1u was determined by western-blot analysis.RESULTS:B19 DNA was detected in 94.6% (35/37) of colon adenocarcinomas, 67.6% (25/37) of adjacent noncancerous tissues, 85.6% (30/35) of polyps, and 60.0% (6/10) of normal controls by nested PCR, respectively. Analysis of the microdissected material confirmed the presence of viral DNA in colonic neoplastic epithelium. ISH detected B19 DNA in 81.1% (30/37) of colon adenocarcinomas, 43.2% (16/37) of adjacent noncancerous tissues, 74.3% (26/35) of polyps, and 50.0% (5/10) of normal controls, respectively. B19 protein VP1/VP2 was found in 78.4% (29/37), 32.4% (12/37), and 57.1% (20/35) of colon adenocarcinomas, tumor-adjacent tissues, and polyps, respectively, but not in normal colons (none of 10). There were significant differences in nested PCR, ISH, and IHC between adenocarcinoma and non-neoplastic adjacent tissues, and between adenocarcinoma and normal controls. Transfection of colon-cancer cells (Lovo) by inducible vector for VP1u resulted in marked upregulation of cyclo-oxygenase-2 proteins.CONCLUSIONS:Parvovirus B19 nucleic acids commonly exist in human colon tissues and VP1/VP2 antigen is preferentially located in colon polyps and adenocarcinomas lesions. B19 viral products VP1u may induce important oncogenic pathways in colon-cancer cells.

Effects of human bone marrow stromal cell line (HFCL) on the proliferation, differentiation and apoptosis of acute myeloid leukemia cell lines U937, HL-60 and HL-60/VCR

This study investigated the effects of human bone marrow fibroblastoid stromal cell line (HFCL) on the proliferation, differentiation and chemosensitivity of acute myeloid leukemia cells (AML) in vitro coculture. By setting up coculture system of sensitive U937, HL-60 cell line and multidrug-resistant (MDR) HL-60/VCR cells in direct contact with human bone marrow fibroblastoid stromal cell line HFCL, or separated by transwell, the proliferation of AML cells cocultured with HFCL cells was inhibited, compared with AML cells alone. And NBT positive cells increased slightly. The percentage of G1 phase cells of AML cells cocultured with HFCL cells was higher than that without HFCL cells, and that of S phase cells was lower. The expression of CD11b and CD14 increased. Meanwhile HL-60 and HL-60/VCR cells treated by TPT were observed to have apoptosis characteristic morphological changes. The proportion of G0/G1 HL-60 and HL-60/VCR cells treated with TPT increased and the sub-G1 increased. The percentage of Annexin V-positive cells and apoptotic cells increased with expression of activated Caspase-3 and the reduced expression of Bcl-2. But when they were cocultured with HFCL cells, the percentage of Annexin V-positive cells and apoptotic cells decreased and sub-G1 reduced. After indirect contact with HFCL cells the expression of activated Caspase-3 decreased and the expression of Bcl-2 increased. After direct contact with HFCL cells for 96 h, the expression levels of 582 genes in HL-60 cells were up-regulated, and 1,323 genes were down-regulated at least twofold by Affymetrix GeneChip Human Genome U133 set A. The expression change in some genes, such as HL14, was confirmed by RT-PCR and northern blot. In a word, HFCL cells could inhibit the proliferation, induce the monocytic differentiation of U937, HL-60 and HL-60/VCR cells, and prevent TPT-induced apoptosis in HL-60 and HL-60/VCR cells via modulation of Bcl-2 and active Caspase-3. Many genes might take part in the influence of HFCL cells on AML cells, which may give important insights into the interaction of bone marrow stromal cells and leukemic cells.

PBK/TOPK Expression During TPA-Induced HL-60 Leukemic Cell Differentiation

OBJECTIVEnThis study concerns expression of PBK/TOPK during differentiation of HL-60 leukemic cells induced by tetradecanoyl phorbol acetate (TPA).nnnMETHODSnWright-Giemsa staining was performed to observe morphological changes in the HL-60 cells, and flow cytometry was used to assess the cell cycle and CD11b, CD14, CD13, and CD33 expression. PBK/TOPK levels were determined by Western blot analysis.nnnRESULTSnAfter treating HL60 cells with 5.1×10⁻⁹ mmol/L of TPA for three days, the number of nitroblue-tetrazolium-positive cells and CD11b, CD13, and CD14 expression increased, whereas the PBK/TOPK levels decreased.nnnCONCLUSIONSnTPA can inhibit proliferation and induce differentiation of HL60 cells of the granulocytic or monocytic lineage. PBK/TOPK expression was downregulated during this process, whereas the Pho-PBK/TOPK expression was increased.