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961.
A multicenter prospective study was performed on 160 asthmatic adults suffering from acute episodes of bronchitis and 88 non-asthmatic controls, to investigate potential associations among Chlamydia pneumoniae infection and/or anti-C. pneumoniae heat shock protein 10 antibodies, and asthma. We used micro-immunofluorescence to detect serum anti-C. pneumoniae IgG, IgA and IgM antibodies and enzyme-linked immunosorbent assay to detect serum anti-Chsp10 peptide IgG antibodies. The serological prevalence of C. pneumoniae was 73.1%. An association was observed between the presence of anti-Chsp10 antibodies and adult onset asthma. The humoral immune responses were not confined to any particular region of the Chsp10 protein. 相似文献
962.
Targeting dendritic cells for priming cellular immune responses 总被引:5,自引:0,他引:5
The cardinal role of dendritic cells (DC) in priming adaptive immunity and in orchestrating immune responses against all classes of pathogens and also against tumors is well established. Their unique potential both to maintain self-tolerance and to initiate protective immune responses against foreign and/or dangerous structures is based on the functional diversity and flexibility of these cells. Tissue DC lining antigenic portals such as mucosal surfaces and the skin are specialized to take up a wide array of compounds including proteins, lipids, carbohydrates, glycoproteins, glycolipids and oligonucleotides, particles carrying such structures and apoptotic or necrotic cells. This process is facilitated by specialized receptors with high endocytic capacity, which provides potential targets for delivering designed molecules. The best route for targeting B- and/or T cell epitopes, however, is still the subject of intense investigation. Immature DC, which reside in various tissues, can be activated by pathogens, stress and inflammation or modified metabolic products, which induce mobilization of cells to draining lymph nodes where they act as highly potent professional antigen presenting cells. This is brought about by the ability to present their accumulated intracellular content for both CD4+ helper (Th) and CD8+ cytotoxic/cytolytic T lymphocytes (Tc/CTL). Engulfed proteins are processed intracellularly and their peptide fragments are transported to the cell surface in the context of major histocompatibility complex encoded class I and II molecules for presentation to Th cells and CTLs, respectively. The T cell priming capacity of DC, however, depends not only on antigen presentation but also on other features of DC. Human monocyte-derived DC provide an excellent tool to study the internalizing, antigen-presenting and T cell-activating functions of DC at their immature and activated differentiation states. These biological activities of DC, however, are highly dependent on their migratory potential from the peripheral non-lymphoid tissues to the lymph nodes, on the expression of adhesion molecules, which support the interaction of DC with T lymphocytes, and the cytokines secreted by DC, which polarize immune responses to Th1-mediated cellular or Th2-mediated antibody responses. These results altogether demonstrate that monocyte-derived DC are useful candidates for in vitro or in vivo targeting of antigens to induce efficient adaptive immune responses against pathogens and also against tumors. 相似文献
963.
Automated whole mount localisation techniques for plant seedlings 总被引:13,自引:0,他引:13
Friml J Benková E Mayer U Palme K Muster G 《The Plant journal : for cell and molecular biology》2003,34(1):115-124
Plant biology is currently experiencing a growing demand for easy and reliable mRNA and protein localisation techniques. Here, we present novel whole mount in situ hybridisation and immunolocalisation protocols, suitable to localise mRNAs and proteins in Arabidopsis seedlings. We demonstrate that these methods can be used in different organs of Arabidopsis seedlings as well as in other plant species. In order to achieve better reproducibility and higher throughput, we modified these protocols for automation to be performed by a liquid handling robot. In addition, we show that other procedures such as reporter enzyme assays and tissue clearing can be similarly automated. We present examples of application of our protocols including mRNA localisation and proteins and epitope tag (co)localisations which demonstrate that these methods provide reliable and versatile tools for expression, localisation and anatomical studies in plants. 相似文献
964.
At five European sites, differing in atmospheric Sinputs by a factor of 6, and differing in S isotope signatures ofthese inputs by up to 14 (CDT), we investigated thedirection and magnitude of an assimilation-related 34S shiftand the relationship between atmospheric deposition and Sretention in selected ecosystem compartments. Bulk precipitationand spruce throughfall were collected between 1994 and 1996 inthe Isle of Mull (Scotland), Connemara (Ireland), Thorne Moors(England), Rybárenská slat' and Oceán (both Czech Republic) andanalyzed for sulfate concentrations and 34S ratios. Eighteenreplicate samples per site of living Sphagnum collected inunforested peatlands and 18 samples of spruce forest floorcollected near each of the peatlands were also analyzed for Sconcentrations and 34S ratios. Assimilation of S was associatedwith a negative 34S shift. Plant tissues systematicallypreferred the light isotope 32S, on average by 2. There wasa strong positive correlation between the non-marine portion ofthe atmospheric S input and total S concentration in forest floorand Sphagnum, respectively (R = 0.97 and R = 0.85). Elevated Sinputs lead to higher S retention in these two organic-richcompartments of the ecosystem. It follows that equal emphasismust be placed on organic S as on adsorption/desorption ofinorganic sulfate when studying acidification reversal inecosystems. The sea-shore sites had rainfall enriched in theheavy isotope 34S due to an admixture of sea-spray. The inlandsites had low 34S reflecting 34S of sulfur emitted from localcoal-burning power stations. Sphagnum had always lower S contentsand higher 34S ratios compared to forest floor. The within-siterange of 34S ratios of Sphagnum and forest floor was wide (upto 12) suggesting that at least six replicate samples shouldbe taken when using 34S as a tracer. 相似文献
965.
Vonasek E Mateu L Luzzati V Marquez G Vargas R Céspedes G Cotúa M Borges J 《European biophysics journal : EBJ》2001,30(1):11-16
A sural nerve dissected from a recently dead patient displayed an unusual X-ray diffraction pattern, suggesting that in situ and at the time of the patient's death the myelin sheaths were in a swollen state. Diffraction patterns of the swollen type were also recorded from: (1) a sural nerve from the corpse of a neurologically healthy person after soaking the nerve with Ringer solution at pH 5.5; (2) sciatic nerves dissected from rat cadavers at increasing time after death. In all the cases the swollen patterns reversed to the native type upon superfusion with Ringer solution at pH 7.3. The postmortem effect is to decrease the pH of the fluids surrounding the nerves in the cadavers. Our experiments show that the early postmortem processes have the effect of acidifying PNS nerves and that as a consequence of acidification the myelin sheaths swell. 相似文献
966.
Glycoproteins, as a class of biomolecules, exhibit much more heterogeneous structures than non-glycosylated proteins. They present a challenging area of research. Model glycoproteins with well-defined protein and carbohydrate structures are helpful in the search for high-resolution methods for the separation of glycoproteins. Neoglycoproteins, maltose-modified chymotrypsin and lactose-modified chymotrypsin, were synthesised by modifying chymotrypsin with maltose and lactose, respectively, using the reductive amination method. Boronate chromatography was applied to isolate the neoglycoproteins from non-glycosylated substances. The use of Tris–HCl as a shielding reagent during the boronate chromatography proved to be efficient in eliminating unwanted interactions between the boronate ligand and the peptide backbone of chymotrypsin. The retention time of neoglycoproteins on the boronate column was increased with increasing the degree of modification. 相似文献
967.
Joe B Remmers EF Dobbins DE Salstrom JL Furuya T Dracheva S Gulko PS Cannon GW Griffiths MM Wilder RL 《Immunogenetics》2000,51(11):930-944
Rat Chromosome 10 (RNO10) harbors Cia5, a non-MHC quantitative trait locus (QTL) that regulates the severity of type II collagen-induced arthritis (CIA) in DAxF344 and DAxBN F2 rats. CIA is an animal model with many features that resemble rheumatoid arthritis. To facilitate analysis of Cia5 independently of the other CIA regulatory loci on other chromosomes, DA recombinant QTL speed congenic rats, DA.F344(Cia5), were generated. These QTL congenic rats have a large chromosomal segment containing Cia5 (interval size < or =80.1 cM) from CIA-resistant F344 rats introgressed into their genome. Phenotypic analyses of these rats for susceptibility and severity of CIA confirmed that Cia5 is an important disease-modifying locus. CIA severity was significantly lower in the Cia5 congenic rats than in DA controls. We also generated DA Cia5 speed sub-congenic rats, DA.F344(Cia5a), which had a smaller segment of the F344 genome, Cia5a, comprising only the distal q-telomeric end (interval size < or = 22.5 cM) of Cia5, introgressed into their genome. DA.F344(Cia5a) sub-congenic rats also exhibited reduced CIA disease severity compared with the parental DA rats. The regulatory effects in both congenic strains were sex influenced. The disease-ameliorating effect of the larger fragment, Cia5, was greater in males than in females, but the effect of the smaller fragment, Cia5a, was greater in females. We also present an improved genetic linkage map covering the Cia5/Cia5a region, which we have integrated with two rat radiation hybrid maps. Comparative homology analysis of this genomic region with mouse and human chromosomes was also undertaken. Regulatory loci for multiple autoimmune/inflammatory diseases in rats (RNO10), mice (MMU11), and humans (HSA17 and HSA5q23-q31) map to chromosomal segments homologous to Cia5 and Cia5a. 相似文献
968.
Macrophage infiltration in human non-small-cell lung cancer: the role of CC chemokines 总被引:8,自引:0,他引:8
Arenberg DA Keane MP DiGiovine B Kunkel SL Strom SR Burdick MD Iannettoni MD Strieter RM 《Cancer immunology, immunotherapy : CII》2000,49(2):63-70
Bronchogenic carcinoma is the leading cause of malignancy-related mortality in the United States, with an overall 5-year
survival rate of less than 15%. This aggressive behavior reflects, among other traits, the capacity of the tumor to evade
normal host immune defenses, and to induce a pro-angiogenic environment. A central feature of any immune response toward tumors
is the recruitment of specific immune cell populations. In the present study we investigated the infiltration of monocytes
in human specimens of non-small-cell lung cancer (NSCLC). The presence of macrophages in NSCLC tumors was documented by immunohistochemistry.
In vitro chemotaxis assays demonstrated higher monocyte chemotactic activity in NSCLC tumor homogenates than in normal lung
tissue. We next investigated the expression of CC chemokines within specimens of NSCLC tumors. Levels of the CC chemokines
were higher in NSCLC tumor tissue than in normal lung tissue. Immunolocalization showed that the cells associated with antigenic
CC chemokines were the malignant tumor cells, as well as occasional stromal cells. Maximal inhibition of monocyte chemotaxis
induced by NSCLC in vitro occurred in the presence of neutralizing antibodies to MCP-1 and MIP-1β. On follow-up of 15 patients
in whom we quantified macrophage infiltration, we found that those with recurrence of disease had higher levels of macrophage
infiltration in their initial tumors. However, the functional significance of CC-chemokine-mediated macrophage infiltration
into NSCLC remains to be determined.
Received: 12 November 1999 / Accepted: 10 December 1999 相似文献
969.
Cancer vaccines: single-epitope anti-idiotype vaccine versus multiple-epitope antigen vaccine 总被引:5,自引:0,他引:5
Maruyama H Zaloudik J Li W Sperlagh M Koido T Somasundaram R Scheck S Prewett M Herlyn D 《Cancer immunology, immunotherapy : CII》2000,49(3):123-132
In this study, we compared the immunogenicity and tumor-protective activity of anti-idiotypic antibodies mimicking a single
tumor-associated epitope and tumor-associated antigen expressing multiple potentially immunogenic epitopes. We focused our
study on the colorectal-carcinoma(CRC)-associated antigen GA733 (also known as CO17-1A/KS1-4/KSA/EpCAM). Monoclonal anti-idiotypic
antibody (Ab2) BR3E4 was produced against murine anti-CRC mAb CO17-1A (Ab1) in rats. Full-length native GA733 protein was
isolated from human tumor cells, and the extracellular domain protein (GA733-2E) was isolated from supernatants of recombinant
baculovirus-infected insect cells by immunoafffinity chromatography. The immunomodulatory activity of the Ab2 was compared
with that of the antigen, both in rabbits and in mice. Mice, like humans but not rabbits, express a GA733 antigen homologue
on some of their normal tissues. Thus, these in vivo models allow the comparison of the immunogenicity of Ab2 and antigen
in the presence (mice) and absence (rabbits) of normal tissue expression and immunological tolerance of the GA733 antigen
homologue. In rabbits, aluminum-hydroxide(alum)-precipitated native GA733 antigen was superior to alum-precipitated Ab2 in
inducing specific humoral immunity. In mice, alum-precipitated recombinant GA733-2E antigen, but not alum-precipitated Ab2,
induced specific humoral immunity. However, when the Ab2 was administered to mice in Freund's complete adjuvant, specific
humoral immune responses were elicited. Ab2 in complete Freund's adjuvant and GA733-2E in alum were compared for their capacity
to induce antigen-specific cellular immunity in mice. Whereas lymphoproliferative responses were obtained with the recombinant
antigen only, delayed-type hypersensitivity responses were obtained with both recombinant antigen and Ab2, although these
responses were lower than after antigen immunization. The recombinant antigen in alum did not protect mice against challenge
with antigen-positive syngeneic murine CRC cells. Similar studies with Ab2 BR3E4 mimicking the CO17-1A epitope were not possible
because the tumor cells do not express this epitope after transfection with the human GA733-2 cDNA. However, similar studies
with Ab2 mimicking the epitope defined by mAb GA733, which is expressed by the transfected tumor cells, indicated a lack of
tumor-protective activity of this Ab2. In contrast, the full-length antigen expressed by recombinant adenovirus inhibited
the growth of established tumors in mice. In conclusion, soluble antigen is a more potent modulator of humoral and cellular
immune responses than Ab2, both administered in adjuvant. However, for induction of protective immunity, the immunogenicity
of the antigen must be further enhanced, e.g., by expression of the antigen in a viral vector.
Received: 27 December 1999 / Accepted: 27 January 2000 相似文献
970.
Behaviour of interleukin-2 serum levels in advanced non-small-cell lung cancer patients: relationship with response to therapy and survival 总被引:8,自引:0,他引:8
Orditura M Romano C De Vita F Galizia G Lieto E Infusino S De Cataldis G Catalano G 《Cancer immunology, immunotherapy : CII》2000,49(10):530-536
Interleukin(IL)-2 is a T helper (Th) 1 type cytokine that has been shown to play an important role in antitumour immune responses.
In this study, the prognostic significance of serum IL-2 levels was investigated in 60 advanced non-small-cell lung cancer
(NSCLC) patients. IL-2 serum levels were determined before chemotherapy, at the end of chemotherapy and during follow-up,
using a commercially available enzyme-linked immunoadsorbent assay kit. The results were analysed according to the response
to therapy and were used to generate a model predicting overall survival and time to treatment failure. All 60 patients were
shown to have higher IL-2 serum levels than controls (P < 0.0001). Stage IV patients had significantly lower IL-2 levels than stage III patients (P < 0.0001), although they were still significantly higher than controls (P < 0.0001). It is interesting that, when patients were divided into responders and non-responders according to the response
to therapy, the former were shown to have significantly higher pre-chemotherapy levels than the latter (P < 0.0001). Moreover, a further significant increase in IL-2 serum levels (P=0.004) and a significant decrease (P < 0.0001) were shown in responders and non-responders, respectively at the end of the therapy. Using univariate and multivariate
analyses, both overall survival and time to treatment failure were shown to be affected by the mean pathological levels of
IL-2. Furthermore, the prognostic significance of the serum level of IL-2 was confirmed by the stepwise regression analysis.
In conclusion, determination of pre-treatment IL-2 serum levels was shown to be of independent prognostic utility in patients
with advanced NSCLC; therefore, its possible use for prediction of outcome is proposed.
Received: 16 March 2000 / Accepted: 27 July 2000 相似文献