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51.
Adult schistosome cDNA libraries as a source of antigens for the study of experimental and human schistosomiasis 总被引:6,自引:0,他引:6
M Knight A J Simpson Q Bickle P Hagan A Moloney A Wilkins S R Smithers 《Molecular and biochemical parasitology》1986,18(2):235-253
Protective immunity has been demonstrated in experimental schistosomiasis and is also believed to occur in man. It can be mediated by antibodies from infected animals or animals immunized with attenuated organisms. Recombinant Escherichia coli synthesizing antigenic polypeptides from the three principal species of schistosome that infect man, Schistosoma mansoni, S. japonicum and S. haematobium, have been constructed. Libraries of adult worm cDNA were prepared from each species in the expression vector lambda gt 11 and directly screened with antibodies from animals experimentally immunized with S. mansoni and S. japonicum and from humans infected with S. haematobium. The S. mansoni clones have been analysed in greatest detail. At least four different types of clones were identified. All the detected recombinant polypeptide antigens were recognised by antibodies from chronically infected mice and most were also recognised by antibodies from mice immunized with attenuated cercariae and anti-surface membrane antibodies. Clones synthesizing species-specific antigens for both S. mansoni and S. japonicum were identified by simultaneous screening of both libraries. At least three types of S. haematobium clones were identified by screening with human infection serum, most of which were species-specific. All the antigens were in the form of fusion peptides with E. coli beta-galactosidase and their expression was induced by isopropylthiogalactopyranoside. Since known protective monoclonal antibodies recognise highly glycosylated membrane proteins which cannot be identified in the form of nascent polypeptides, the direct identification of polypeptide antigens defined by their reactivity, as reported here, is an essential step in producing reagents by recombinant DNA technology, suitable for vaccination and diagnosis. 相似文献
52.
Intensive immunosuppression in patients with disseminated sclerosis. III. Lymphocyte response in vitro. 总被引:2,自引:0,他引:2
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S C Knight E M Lance J Abbosh A Munro J O''''Brien 《Clinical and experimental immunology》1975,21(1):23-31
Lymphocytes from fifteen multiple sclerosis patients gave responses to phytohaemagglutinin (PHA), conconavalin A (con A) and pokeweed mitogen (PWM) which were in the normal range. However, the responses of lymphocytes to stimulation by an allogeneic lymphoid cell line (LCL) were significantly lower in HLA-7-positive than in HLA-7-negative patients (a distinction not found in control groups). Depression of con A, PHA and PWM responses were observed during intensive immunosuppression. Responses to LCL were unaltered or increased during initial azathioprine and prednisone treatment. The depression of this response following antilymphocyte globulin (ALG) treatment was delayed in the HLA-7-positive patients. One week after the end of ALG treatment, most PHA, con A and PWM responses had returned to low normal values. Reduction of azathioprine and prednisone treatment at the end of 1 year resulted in a sharp rise in PHA and con A responses in some patients. Relapses in patients were frequently associated with low responses to LCL cells. 相似文献
53.
The interaction of normal lymphocytes and cells from lymphoid cell lines: II. Variations relating to cell line source and lymphocyte donor
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Irradiated LCL cells from several sources activated the blood lymphocytes of a panel of donors. Individual differences were present but were small. Stimulated blood lymphocytes were more potent activators (after X-irradiation) than small lymphocytes in a one-way mixed lymphocyte reaction, indicating that the state of metabolic activity of the `stimulating' lymphocyte affects the level of activation achieved. However, LCL cells incubated for several days after irradiation had not lost stimulatory capacity. Lymphocytes of pigs, rabbits, rats and mice were much less responsive than human lymphocytes. The response of animal lymphocytes generally varied with the species of origin of the serum used in the cultures. 相似文献
54.
Peripheral blood and synovial fluid T cells differ in their response to alloantigens and recall antigens presented by dendritic cells.
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A J Stagg B Harding R A Hughes A Keat S C Knight 《Clinical and experimental immunology》1991,84(1):72-77
Properties of T cells from inflammatory lesions were analysed by comparing the response of peripheral blood (PB) and synovial fluid (SF) T cells from 19 patients with a range of arthropathies to enriched allogeneic dendritic cells (DC) in a primary mixed leucocyte reaction (MLR). In 17 patients the proliferative response of SF T cells was significantly (P less than 0.05) less than that of PB lymphocytes. The reduced response of SF T cells was observed in all disease categories studied and could not be attributed to differences in cell number requirements or response kinetics. Addition of recombinant interleukin-2 enhanced the response of SF T cells in a dose-dependent manner. Cell mixing experiments suggested that active suppression was not the underlying mechanism of the poor MLR response of SF T cells. In contrast to the MLR response. SF T cells were able to mount vigorous proliferative responses to recall antigen presented by autologous antigen-presenting cells. The possibility is discussed that T cells compartmentalized at inflammatory lesions are a unique population with a diminished ability to interact with DC and respond to primary stimuli but an ability to respond to secondary antigenic challenge. 相似文献
55.
Adhesion molecules are upregulated on dendritic cells isolated from human blood. 总被引:2,自引:0,他引:2
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This study investigated whether the high expression of adhesion molecules on enriched preparations of circulating dendritic cells (DCs) was an intrinsic property of the cells or whether it was a consequence of the procedure used to isolate them from blood. Expression of the beta 1, beta 2 integrins (CD11/CD18 family) and other adhesion molecules on DCs in whole blood was compared with that on isolated DCs. Dendritic cells were identified by flow cytometry as leucocytes that were positive for human leucocyte antigen (HLA)-DR, but negative for CD3, CD14, CD16, CD19 and CD56. In contrast to a minority of DCs in whole blood, the majority of isolated DCs expressed the beta 2 integrins and there were a greater number of cells bearing CD44, CD54 and some of the beta 1 integrins (notably CD49b, CD49d, CD49e and CD29). An increase in the proportion of DCs bearing adhesion molecules was generally apparent at the isolation stage when mononuclear cells, which had been incubated overnight, were centrifuged on a metrizamide gradient to enrich for cells of low density. Inclusion of an inhibitor of protein glycosylation and exocytosis (brefeldin A) at all stages of separation partially prevented an increase in the percentage of DCs bearing CD18, C29 and C54 whereas the inclusion of cycloheximide (an inhibitor of polypeptide synthesis) interfered with increases in the percentage of cells bearing CD29 and CD54. Neither of these antagonists had an effect on the intensity of adhesion molecule expression. We suggest that some of the adhesion-dependent functions of isolated DCs are caused, in part, by an upregulation of surface adhesion molecules induced by the enrichment procedure. 相似文献
56.
The in vitro transformation of thymocytes and lymphocytes from humans, rabbits and guinea-pigs and from thymomas 总被引:1,自引:5,他引:1
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Stella Knight J. Bradley J. J. Oppenheim N. R. Ling 《Clinical and experimental immunology》1968,3(4):323-341
The response of human, rabbit and guinea-pig thymocytes in culture to blastogenic stimulants such as phytohaemagglutinin (PHA) and staphylococcal filtrate (SF) was usually considerably less than that seen with equivalent numbers of peripheral blood and lymph node lymphocytes. 相似文献
57.
PCR strategy for identification and differentiation of small pox and other orthopoxviruses. 总被引:4,自引:0,他引:4
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S L Ropp Q Jin J C Knight R F Massung J J Esposito 《Journal of clinical microbiology》1995,33(8):2069-2076
Rapid identification and differentiation of orthopoxviruses by PCR were achieved with primers based on genome sequences encoding the hemagglutinin (HA) protein, an infected-cell membrane antigen that distinguishes orthopoxviruses from other poxvirus genera. The initial identification step used a primer pair of consensus sequences for amplifying an HA DNA fragment from the three known North American orthopoxviruses (raccoonpox, skunkpox, and volepox viruses), and a second pair for amplifying virtually the entire HA open reading frame of the Eurasian-African orthopoxviruses (variola, vaccinia, cowpox, monkeypox, camelpox, ectromelia, and gerbilpox viruses). RsaI digest electropherograms of the amplified DNAs of the former subgroup provided species differentiation, and TaqI digests differentiated the Eurasian-African orthopoxviruses, including vaccinia virus from the vaccinia virus subspecies buffalopox virus. Endonuclease HhaI digest patterns distinguished smallpox variola major viruses from alastrim variola minor viruses. For the Eurasian-African orthopoxviruses, a confirmatory step that used a set of higher-sequence-homology primers was developed to provide sensitivity to discern individual virus HA DNAs from cross-contaminated orthopoxvirus DNA samples; TaqI and HhaI digestions of the individual amplified HA DNAs confirmed virus identity. Finally, a set of primers and modified PCR conditions were developed on the basis of base sequence differences within the HA genes of the 10 species, which enabled production of a single DNA fragment of a particular size that indicated the specific species. 相似文献
58.
A 63-year-old man with iron loss anaemia and hypercalcaemia was found to have a renal cell carcinoma. Despite the iron-deficient blood and bone marrow picture, the serum ferritin concentration was markedly raised. This was mainly due to a “basic isoferritin”. The serum parathormone concentration was normal. The serum ferritin and calcium concentrations returned to normal after the tumour was removed. We propose that the renal cell carcinoma cells in this patient secreted the basic isoferritin as well as humoral factor(s) responsible for hypercalcaemia. 相似文献
59.
Gammadelta T cells are expanded in human IgA nephropathy and in a rat model of adriamycin (ADR)-induced nephropathy. Despite different diseases and species, these renal gammadelta T cells use a restricted set of gammadelta T cell receptor (TCR) genes. To explore whether this phenomenon of post injury expansion of gammadelta T cells occurs in autoimmune-mediated glomerulonephritis, we studied gammadelta TCR genes in Heymann nephritis (HN). Gammadelta T cells were increased in HN kidneys (p<0.001). These gammadelta T cells predominantly expressed Vgamma6/Vdelta1 genes and used canonical matching sequences previously seen in the other models of renal injury. Gammadelta T cells from the kidneys expressed high levels of TGF-beta, IL-4 and IL-5. The gammadelta T cells from both ADR-treated and HN kidneys expressed NKG2D, the NK cell-activating receptor. These results demonstrate that the majority of gammadelta T cells in the HN kidney use a canonical Vgamma6/Vdelta1 TCR--the gammadelta TCR previously described in the rat ADR-treated kidney. The restriction in gammadelta TCR seen in two completely different models of kidney injury and the expression of an innate activating molecule NKG2D suggests that the gammadelta T cells may be responding to tissue stress from injury and producing a regulatory response. 相似文献
60.
Primary proliferative and cytotoxic T-cell responses to HIV induced in vitro by human dendritic cells. 总被引:8,自引:0,他引:8
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In earlier studies, primary proliferative and cytotoxic T-cell (CTL) responses to influenza virus were produced in vitro by using mouse dendritic cells (DC) pulsed with virus or viral peptide as the stimulus for syngeneic T cells in 20-microliters hanging-drop cultures. We have now adapted this system for producing primary responses with cells from non-immune donors to produce primary proliferative and CTL responses to human immunodeficiency virus I (HIV) and to HIV peptides in vitro using cells from normal human peripheral blood. All donors in this study were laboratory personnel with no history of HIV infection. DC enriched from peripheral blood were exposed to HIV in vitro and small numbers were added to T lymphocytes in 20-microliters hanging drops. Proliferative responses to virus-infected DC were obtained after 3 days in culture. After 6 days, CTL were obtained that killed virus-infected autologous--but not allogeneic--phytohaemagglutinin (PHA)-stimulated blast cells. Proliferative and CTL responses were obtained using cells from 14 random donors expressing a spectrum of major histocompatibility complex (MHC) types but the CTL, once produced, showed killing restricted by the MHC class I type. Treatment of cultures with monoclonal antibody (mAb) to CD4-positive cells at the beginning of culture blocked the development of both proliferative and CTL responses, but treatment after 5 days had no effect on the CTL activity. Treatment with MCA to CD8-positive cells at the beginning of culture did not block proliferation significantly, but treatment either before or after the 5-day culture period blocked CTL responses. Collaboration between proliferating CD4-positive cells and CD8-positive cells may thus be required to produce CTL of the CD8 phenotype. DC exposed to HIV also produced CTL that killed autologous blast cells pulsed with gp120 envelope glycoprotein. However, DC infected with whole virus did not produce CTL that lysed target cells pulsed with a synthetic peptide, which included a known T-cell epitope of gp120 (representing amino acids 111-126). DC pulsed with gp120 were a poor stimulus for the development of CTL. In contrast, DC pulsed with the peptide (111-126) stimulated both proliferative and CTL responses. The latter killed not only target cells pulsed with the peptide itself or with gp120 but also killed virus-infected autologous blast cells. CTL were again obtained reproducibly with this peptide using donors expressing a spectrum of MHC types.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献