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L Wen

Publications and source records attributed to L Wen.

139 records · Page 8Linked to original sources
PubMed1989

Purification and characterization of adult diarrhea rotavirus: identification of viral structural proteins.

Adult diarrhea rotavirus (ADRV) is a newly identified strain of noncultivable human group B rotavirus that has been epidemic in the People's Republic of China since 1982. We have used sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western (immuno-) blot analysis to examine the viral proteins present in the outer and inner capsids of ADRV and compared these with the proteins of a group A rotavirus, SA11. EDTA treatment of double-shelled virions removed the outer capsid and resulted in the loss of three polypeptides of 64, 61, and 41, kilodaltons (kDa). Endo-beta-N-acetylglucosaminidase H digestion of double-shelled virions identified the 41-kDa polypeptide as a glycoprotein. CaCl2 treatment of single-shelled particles removed the inner capsid and resulted in the loss of one polypeptide with a molecular mass of 47 kDa. The remaining core particle had two major structural proteins of 136 and 113 kDa. All of the proteins visualized on sodium dodecyl sulfate-polyacrylamide gel electrophoresis were antigenic by Western blot analysis when probed with convalescent-phase human and animal antisera. A 47-kDa polypeptide was most abundant and was strongly immunoreactive with human sera, animal sera raised against ADRV and against other group B animal rotaviruses (infectious diarrhea of infant rat virus, bovine and porcine group B rotavirus, and bovine enteric syncytial virus) and a monoclonal antibody prepared against infectious diarrhea of infant rat virus. This 47-kDa inner capsid polypeptide contains a common group B antigen and is similar to the VP6 of the group A rotaviruses. Human convalescent-phase sera also responded to a 41-kDa polypeptide of the outer capsid that seems similar to the VP7 of group A rotavirus. Other polypeptides have been given tentative designations on the basis of similarities to the control preparation of SA11, including a 136-kDa polypeptide designated VP1, a 113-kDa polypeptide designated VP2, 64- and 61-kDa polypeptides designated VP5 and VP5a, and several proteins in the 110- to 72-kDa range that may be VP3, VP4, or related proteins. The lack of cross-reactivity on Western blots between antisera to group A versus group B rotaviruses confirmed that these viruses are antigenically quite distinct.

Adult↗
PubMed1989

High level, cell-specific expression of ornithine decarboxylase transcripts in rat genitourinary tissues.

We evaluated transcript levels for the rate-limiting enzyme in polyamine biosynthesis, ornithine decarboxylase (ODC), in rat tissues by Northern blotting and in situ hybridization histochemistry, using a rat cDNA probe. ODC transcripts were expressed at a high level, relative to levels in other tissues, in the kidney and testis of the adult rat; maximal levels of transcripts in these tissues occurred after sexual maturation had taken place, i.e. between 20 and 150 days of age. In situ hybridization histochemistry revealed high level expression in the kidney, testis, prostate, and seminal vesicles of the male rat; this high level expression was limited to certain cell types: kidney, S3 cells of the proximal convoluted tubule; prostate and seminal vesicles, glandular or luminal epithelial cells; and testis, early spermatogenic cells. High level expression of ODC mRNA disappeared from the prostate and seminal vesicle epithelial cells after castration and reappeared with testosterone treatment; in contrast, levels of kidney ODC mRNA were essentially unchanged by castration and were similar in male and female adult rats. We conclude that high level ODC mRNA expression occurs in specific cell types in the adult rat, where it appears to be regulated by both androgen-dependent and independent mechanisms.

Animals↗
PubMed1988

Quantitative analysis of precursors frequency of rheumatoid factor (RF) producing human B cells.

Recently, a new culture system has been devised which leads to activation, proliferation and differentiation into antibody secreting cells of at least 90% of human peripheral blood B-lymphocytes. The system uses mutant EL-4 thymoma cells of mouse origin for B cell activation and T cell/macrophage supernatant as source of cytokines (L. Wen et al., Eur. J. Immunol. 17, 887, 1987). By an ELISA system with Fc fragments as antigen and the F(ab)2 fragment of antibodies against human IgM labelled with alkaline phosphatase, we analysed the frequencies of B cells producing IgM-RFs. The mean frequency of IgM-RF producing cells in normal controls was 1/3100 (s.d. = 0.2 log; n = 5) of circulating B cells. In 2 patients with seropositive rheumatoid arthritis (RA) the frequencies were higher (1/492; 1/262), but in 2 other seropositive patients a normal (1/5540) or even a decreased (1/20000) frequency was observed. The possible relation between circulating and synovial RF producing B cells is currently being investigated.

B-Lymphocytes↗
PubMed1987

Limiting dilution assay for human B cells based on their activation by mutant EL4 thymoma cells: total and antimalaria responder B cell frequencies.

The analysis of human B cell responses at the clonal level (limiting dilution assay) is still technically difficult. In the present study we report on a culture system that leads to activation, proliferation and differentiation into antibody-secreting cells (ASC) of about 90% of B cells from peripheral blood or spleen. In this system, B cells are cultured in the presence of a mutant subclone of the mouse thymoma EL4 for B cell activation and human T cell plus macrophage supernatant as source of proliferation and differentiation factors. ASC precursors generating clonal responses of IgM only, IgM plus IgG, or IgG only occurred at a ratio of about 6:3:1. The mean clone size was 380 cytoplasmic Ig+ cells; the mean amount of Ig secreted per clone was 20 ng. Furthermore, it has been found using this system that a considerable proportion of peripheral blood B cells from individuals with a history of malaria infection could generate clones of anti-malaria (Plasmodium falciparum) ASC (range of 0.1 to 1%, n = 6). In a control group of blood donors the corresponding frequencies were 10 times lower (range of 0.01 to 0.1%, n = 9). These results show that the EL4 culture system can be applied to the investigation of the human B cell specificity repertoire and of priming effects such as result from infectious disease.

Adult↗
PubMed1987

Theoretical and practical aspects of B-cell activation: murine and human systems.

We have reviewed observations which were made during studies of murine and human B-cell responses in vitro. One currently faces difficulties in drawing any clear schema as to which external signals elicit which responses (activation, proliferation, differentiation) in B cells. However, the most potent antigen-dependent or polyclonal B-cell responses in vitro occur when, in addition to various cytokines, accessory cells, serum etc., the cultures contain either a) intact T-helper cells which enter into cell-to-cell contact with B cells, or b) some B-cell "mitogen" (T-independent antigen). Murine B cells activated with LPS and anti-Ig antibodies represent a model for the study of IL2 receptor expression and function. LPS does not act on human B cells. Certain mutant EL-4 thymoma cells are potent activators of murine and human B cells via a direct cell-to-cell interaction. The majority of human B cells can be induced to proliferate and generate a few hundred antibody-secreting cells each in the presence of such thymoma cells and a mixture of cytokines. From a practical point of view, this observation should be useful in a variety of investigations such as the analysis of the human B-cell specificity repertoire.

Animals↗
PubMed1986

Protein kinase C-stimulated phosphorylation in vitro of a Mr 80,000 protein phosphorylated in response to phorbol esters and growth factors in intact fibroblasts. Distinction from protein kinase C and prominence in brain.

In previous studies in intact 3T3-L1 fibroblasts and adipocytes, we demonstrated that the phosphorylation state of an acidic, multicomponent Mr 80,000 protein appeared to be a specific and useful marker for the activation state of protein kinase C (Blackshear, P.J., Witters, L.A., Girard, P.R., Kuo, J.F., and Quamo, S.N. (1985) J. Biol. Chem. 260, 13304-13315). In the present studies, we demonstrate that the Mr 80,000 protein from rat adipose tissue was a substrate for protein kinase C in vitro, and co-migrated on two-dimensional gels with the analogous protein from murine 3T3-L1 adipocytes labeled by exposure of intact cells to 32Pi and phorbol 12-myristate 13-acetate. Partial proteolytic maps of the two 32P-proteins were nearly identical, supporting the postulate that the sites phosphorylated by protein kinase C in vitro, and in response to phorbol 12-myristate 13-acetate in vivo, were similar or identical. Despite their similar apparent molecular weights, we were able to distinguish between the Mr 80,000 protein and protein kinase C by several physical criteria. The Mr 80,000 protein kinase C substrate was found in fractions of all rat tissues examined, but was most prominent in rat brain. Phorbol 12-myristate 13-acetate also stimulated phosphorylation of the Mr 80,000 protein in several types of cultured neuronal cells, suggesting a possible role for this protein in cholinergic neurotransmission. The Mr 80,000 protein appears to be a useful marker for protein kinase C activation in a variety of cell types.

Adipose Tissue↗
PubMed1983

Ionic interactions between proteins in nonequilibrium pH gradient electrophoresis: histones affect the migration of high mobility group nonhistone chromatin proteins.

In two-dimensional gel electrophoresis of the high mobility group (HMG) proteins, it has proved necessary to use nonequilibrium pH gradient electrophoresis (NEPHGE) in the first dimension rather than isoelectric focusing, because of the basic character of most of the HMG proteins [D. Tyrell, P. J. Isackson, and G. R. Reeck (1982) Anal. Biochem. 119, 433-439]. In this paper it is reported that in samples that contain histones, the mobilities of HMG proteins (particularly HMG-1, HMG-2, and HMG-E) are severely distorted in NEPHGE. This presumably results from formation of complexes between histones and HMG proteins through ionic interactions. Analysis of HMG proteins by NEPHGE/sodium dodecyl sulfate-gel electrophoresis is thus precluded in samples containing histones. Our results raise the possibility of similar artifacts occurring in NEPHGE (or isoelectric focusing) analysis of other proteins with regions of high charge density.

Animals↗
PubMedDate not supplied

Alphamethrin-impregnated bed nets for malaria and mosquito control in China.

A community-based intervention trial was carried out to evaluate the effectiveness of alphamethrin-impregnated bed nets for control of Plasmodium vivax malaria and its vector in an area of moderate endemicity in southern Henan province, central China in 1990. Malaria incidence was significantly lower in the intervention group than in the comparison group (2.03 vs. 3.57 per 100 person-years at risk). The protective efficacy for malaria incidence was 43%. The prevalence of malaria parasitaemia among children under 10 years old in the intervention group was about one-quarter of that in the comparison group (0.93% vs. 3.25% and 0.71% vs. 1.96% after one and 4 months use of impregnated nets, respectively). Alphamethrin-impregnated bed nets had a mass killing effect on vector mosquitoes. The outdoor person-biting density of Anopheles anthropophagus and A. sinensis decreased by 70.3% and 29.3% respectively. The density of these 2 mosquito species found resting inside treated nets was close to zero. No side effect was found among users of impregnated bed nets. Impregnation with alphamethrin was more effective on polyester than on cotton netting and residual effects lasted at least one year. Use of alphamethrin is less expensive than permethrin and deltamethrin.

Animals↗
PubMedDate not supplied

T-cell-directed hepatocyte damage in autoimmune chronic active hepatitis.

To investigate the function of activated T lymphocytes in autoimmune chronic active hepatitis, 7 of 15 T-cell clones from the peripheral blood of 8 patients were studied. These clones showed specificity for liver-membrane antigen with proliferation when stimulated by rabbit liver cell membranes. 6 of these clones reacted with liver-specific lipoprotein complex, and 1 clone (and 3 subclones) responded to the asialoglycoprotein receptor (ASGPR), both known targets of immune attack in autoimmune chronic active hepatitis. 2 of these clones stimulated autologous B lymphocytes to produce liver-membrane-specific autoantibodies and antibody to the ASGPR. These results suggest that liver-membrane-specific activated T lymphocytes in peripheral blood may be important in the autoimmune attack of chronic active hepatitis.

Adolescent↗