bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression, purification and immunological characterization of the transforming protein E7, from cervical cancer-associated human papillomavirus type 16
Authors: FERNANDO G J P , MURRAY B , ZHOU J , FRAZER I H 
Published: March 2003

SUMMARY
E7 is the major oncogenic protein produced in cervical cancer-associated human papillomavirus type 16 (HPV16). This protein was expressed in Escherichia coli as a glutathione-S-transferase (GST) fusion protein. E7-enriched inclusion bodies were collected from bacterial lysates, were solubilized in 10 m urea, and the protein was purified using anion exchange column chromatography. After removal of endotoxin with serial Triton X-114 extractions, material of high purity (about 90%) was obtained, which is suitable for use in a human clinical trial. This material was immunogenic, and when used as a vaccine, protected mice against challenge with an HPV16 E7 DNA transfected tumour cell line. Based on this observation, the E7GST fusion protein is currently being used in a human clinical trial of a vaccine against HPV16-induced cervical cancer. This fusion protein could be cleaved with thrombin to remove the GST fusion part and further purified by preparative SDS gel electrophoresis to obtain free E7 with &gt; 98% purity.
Intracellular interferon-gamma (IFN-γ) production in normal children and children with atopic dermatitis
Authors: CAMPBELL D E , FRYGA A S , BOL S , KEMP A S 
Published: March 2003

SUMMARYA reduction in the in vitro production of IFN-γ has been consistently described in atopic dermatitis (AD). Whether this reduction is due to a decrease in the population of peripheral blood mononuclear cells (PBMC) producing IFN-γ or reduced IFN-γ production per cell, or a combination of both is not clear. We have examined the intracellular production of IFN-γ in children with AD and in healthy non-atopic controls. As Staphylococcus aureus colonization is a feature of childhood AD, and is postulated to contribute to the cutaneous inflammation in atopic dermatitis, S. aureus and Staphylococcal enterotoxin B (SEB) were used to activate PBMC. Stimulated PBMC from subjects with AD had significantly fewer IFN-γ-containing cells in response to SEB (P &lt; 0.001) and S. aureus (P &lt; 0.01) than normal non-atopic children. In addition, SEB-stimulated PBMC from children with AD had less IFN-γ per cell than normal non-atopic children (P &lt; 0.01). Reduction in the proportion of cells containing IFN-γ was seen in CD4+, CD8+ and natural killer (NK) cells in PBMC from children with AD. Our findings indicate that reduced production of IFN-γ observed in childhood AD is due to both a decrease in the number of IFN-γ-producing cells and a reduced amount of IFN-γ production per cell. Furthermore, we found that this defect was not confined to CD4+ T cells, suggesting a more generalized defect in IFN-γ production in childhood AD.
Reduced naive and increased activated CD4 and CD8 cells in healthy adult Ethiopians compared with their Dutch counterparts
Authors: MESSELE T , ABDULKADIR M , FONTANET A L , PETROS B , HAMANN D , KOOT M , ROOS M T L , SCHELLEKENS P T A , MIEDEMA F , RINKE DE WIT T F 
Published: March 2003

SUMMARYTo assess possible differences in immune status, proportions and absolute numbers of subsets of CD4+ and CD8+ T cells were compared between HIV− healthy Ethiopians (n = 52) and HIV− Dutch (n = 60). Both proportions and absolute numbers of naive CD4+ and CD8+ T cells were found to be significantly reduced in HIV− Ethiopians compared with HIV−Dutch subjects. Also, both proportions and absolute numbers of the effector CD8+ T cell population as well as the CD4+ CD45RA− CD27− and CD8+ CD45RA− CD27− T cell populations were increased in Ethiopians. Finally, both proportions and absolute numbers of CD4+ and CD8+ T cells expressing CD28 were significantly reduced in Ethiopians versus Dutch. In addition, the possible association between the described subsets and HIV status was studied by comparing the above 52 HIV− individuals with 32 HIV+ Ethiopians with CD4 counts &gt; 200/μl and/or no AIDS-defining conditions and 39 HIV+ Ethiopians with CD4 counts &lt; 200/μl or with AIDS-defining conditions. There was a gradual increase of activated CD4+ and CD8+ T cells, a decrease of CD8+ T cells expressing CD28 and a decrease of effector CD8+ T cells when moving from HIV− to AIDS. Furthermore, a decrease of naive CD8+ T cells and an increase of memory CD8+ T cells in AIDS patients were observed. These results suggest a generally and persistently activated immune system in HIV−Ethiopians. The potential consequences of this are discussed, in relation to HIV infection.
Imbalance of IL-1β and IL-1 receptor antagonist mRNA in liver tissue from hepatitis C virus (HCV)-related chronic hepatitis
Authors: GRAMANTIERI L , CASALI A , TRERÈ D , GAIANI S , PISCAGLIA F , CHIECO P , COLA B , BOLONDI L 
Published: March 2003

SUMMARYIncreased levels of IL-1β and IL-1 receptor antagonist (IL-1Ra) have been found in serum of patients with chronic liver diseases, although their expression in liver tissue has not been extensively investigated. The aim of this study was therefore to examine the relationship between IL-1β and IL-1Ra at tissue level in patients with HCV-related chronic active hepatitis (CAH) of varying degrees of severity. IL-1β and IL-1Ra mRNA expression was investigated by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) in 31 patients with CAH of varying severity (classified as minimal/mild in 13 cases and moderate/severe activity in 18 cases) and in 12 control subjects. Quantitative evaluation of IL-1β and IL-1Ra corresponding bands was performed by densitometric image analysis, and expressed in arbitrary units. The 12 controls expressed a similar pattern with a mean IL-1β/IL-1Ra ratio of 1.03 (1.03 ± 0.15 (mean ± s.e.m.), median 0.92, range 0.71–1.45). Minimal/mild activity CAH showed a prevalence of IL-1Ra mRNA expression (1.14 ± 0.64, median 0.43, range 0–8.75) when compared with controls (0.27 ± 0.04, median 0.23, range 0.11–0.45) and with moderate/severe activity CAH (0.20 ± 0.04, median 0.12, range 0–0.67; P = 0.01). Since IL-1β expression was similar in the three groups, a significantly different IL-1β/IL-1Ra ratio emerged between controls, patients with moderate/severe CAH (2.22 ± 0.48, median 2.76, range 0–6.12) and those with minimal/mild activity CAH (0.62 ± 0.15, median 0.5, range 0–1.58, P = 0.005). Patients with higher grades of fibrosis showed a higher IL-1β/IL-1Ra ratio (2.49 ± 0.56, median 2.15, range 0.35–6.12) in comparison with lower grade fibrosis (1.06 ± 0.30, median 0.59, range 0.03–4.50) and control patients (P = 0.01). These results suggest that an imbalance between IL-1β and IL-1Ra, at the tissue level, may contribute to the pathogenesis and the activity of chronic active hepatitis C.
Superantigen-induced T cell responses in acute rheumatic fever and chronic rheumatic heart disease patients
Authors: BHATNAGAR A , GROVER A , GANGULY N K 
Published: March 2003

SUMMARYCD4+ and CD8+ T cells from healthy donors, acute rheumatic fever (ARF) and chronic rheumatic heart disease (CRHD) patients responded variably to a superantigen from Streptococcus pyogenes—Streptococcal pyrogenic erythrogenic toxin A (SPE-A). In vitro culture of CD4+ T cells from ARF patients (CD4-ARF) with SPE-A exhibited a Th1 type of response as they produced high levels of IL-2, while CD4+ T cells from CRHD patients (CD4-RHD) secreted IL-4 and IL-10 in large amounts, i.e. Th2 type of cytokine profile. The skewing of human CD4+ T cells (in response to SPE-A stimulation) to Th1 or Th2 type reflects the role of the two subsets in a disorder with differing intensities at the two extremes of the spectrum. Moreover, the anergy induction experiments revealed that CD8-ARF and CD8-RHD undergo anergy (to different extents), whereas CD4+ T cells do not, in response to re-stimulation by SPE-A. These results initially demonstrate that both CD4+ and CD8+ T cells respond differentially to SPE-A, and hence it is an important observation with respect to the pathogenesis of ARF/CRHD. Anergy in CD8+ T cells in the presence of SPE-A in vitro goes a step further to show the clinical relevance of these cells and their possible role in suppression of the disease.
Early reduction of the over-expression of CD40L, OX40 and Fas on T cells in HIV-1 infection during triple anti-retroviral therapy: possible implications for lymphocyte traffic and functional recovery
Authors: SOUSA A E , CHAVES A F , DOROANA M , ANTUNES F , VICTORINO R M M 
Published: March 2003

SUMMARYFas, CD40L and OX40 are members of the tumour necrosis factor (TNF) receptor superfamily with critical roles in T cell activation and death, B cell function, dendritic cell maturation and leucocyte traffic regulation. The aim of this study was to evaluate the effects of anti-retroviral therapy (HAART) on CD40L, OX40 and Fas expression on freshly isolated peripheral blood T cells by three-colour flow cytometry and compare them with lymphoproliferative responses, peripheral blood cell counts and viral load. Fourteen asymptomatic HIV-1+ patients treated with Lamivudine, Stavudine and Nelfinavir were prospectively investigated sequentially for 48 weeks. At baseline, patients exhibited significantly enhanced proportions and counts of CD40L+ and OX40+ cells within the CD4 subset which were corrected by weeks 8–16 of HAART. Interestingly, in the five patients showing viral load rebound during therapy in spite of increasing CD4 counts, the reduction of the levels of these costimulatory molecules was similarly maintained. Therapy induced a decrease in the over-expression of Fas, particularly in the CD4 subset where normal levels were reached at week 8. This reduction occurred in parallel with the major recovery of lymphoproliferative responses. Higher basal levels and lower reduction of Fas were associated with suboptimal suppression of viraemia. In conclusion, this previously undescribed increased expression of CD40L and OX40 may play a role in the HIV-associated pan-immune activation and represent a possible target for immunointervention, as suggested for several immunologically mediated diseases. Moreover, HAART induced an early correction of the over-expression of Fas, CD40L and OX40 in CD4 T cells which could be involved in the recovery of the cell traffic disturbances and in the T cell renewal capacity.
Studies of the mechanism of cytolysis by tumour-infiltrating lymphocytes
Authors: HISHII M , KURNICK J T , RAMIREZ-MONTAGUT T , PANDOLFI F 
Published: March 2003

AbstractIn order to determine the mechanism of tumour destruction by tumour-infiltrating lymphocytes (TIL), we examined the ability of both CD4+ and CD8+ effector TIL, and TIL clones, to manifest granzyme-mediated and Fas-mediated destruction of tumour targets. In many in vitro studies TIL have been shown to manifest anti-tumour reactivity, yet many tumours escape immunological destruction. To investigate the role of Fas expression and the concomitant sensitivity to the inducibility of apoptotic death, we derived TIL from four melanomas and one glioma. The glioma, and all but one of the melanomas, expressed Fas, but Fas-mediated apoptosis could only be detected if the targets were treated with cyclohexamide. The melanomas and the glioma all expressed detectable cytoplasmic Bcl-2 protein, known to exert anti-apoptotic activity. Lysis of tumours by CD8-enriched cultures and CD8+ clones was Ca2+-dependent and could not be modified by an anti-Fas MoAb. In CD4-enriched cultures or CD4+ clones with cytotoxic potential against tumour cells, cytotoxicity was also Ca2+-dependent. As Ca2+-dependent cytotoxicity is usually the result of secretion of perforin/granzyme-B, we investigated the presence of perforin in cytotoxic CD4+ clones and demonstrated the presence of granular deposits of this enzyme in some of the CD4+clones. Although an anti-Fas MoAb did not block the lysis of melanoma targets by CD4+ clones, the examination of Fas-dependent targets demonstrated that these clones also had the potential to kill by the Fas/Fas ligand system. These data suggest that the predominant mechanism in tumour killing by TIL appears to be perforin–granzyme-dependent, and that the solid tumour cell lines we studied are less susceptible to Fas-mediated apoptosis. As non-apoptotic pathways may enhance tumour immunogenicity, exploitation of the perforin–granzyme-dependent cytotoxic T lymphocyte (CTL) pathways may be important for achieving successful anti-tumour responses.
Elevated serum levels of soluble membrane cofactor protein (CD46, MCP) in patients with systemic lupus erythematosus (SLE)
Authors: KAWANO M , SEYA T , KONI I , MABUCHI H 
Published: March 2003

AbstractMembrane cofactor protein (MCP, CD46) is a cell surface complement regulatory protein which acts as a cofactor for the factor I-mediated cleavage of the activated complement components C3b/C4b. To evaluate the clinical usefulness of serum soluble CD46 as a marker of disease activity in patients with SLE, serum levels of sCD46 were measured by ELISA, using two MoAbs (M160 and M177), each of which recognized two different epitopes on CD46 molecule in SLE, other autoimmune diseases and healthy controls. Serum sCD46 levels in active SLE patients (30.5 ± 14.1 ng/ml) were significantly higher than those of inactive SLE (5.8 ± 7.1 ng/ml; P = 0.0003), rheumatoid arthritis (14.9 ± 11.6 ng/ml; P = 0.0218), primary Sjögren's syndrome (12.3 ± 11.6 ng/ml; P = 0.0039) and normal controls (7.3 ± 3.6 ng/ml; P = 0.0005). The elevated serum sCD46 levels in active SLE patients significantly decreased from 30.5 ± 14.1 ng/ml to 8.0 ± 6.3 ng/ml after effective corticosteroid and immunosuppressant therapy (P = 0.018). Additionally, we found a significant negative association between increasing concentration of sCD46 and decreasing levels of CH50 in SLE (r = –0.598, P = 0.0009). These results suggest that sCD46 reflects in vivo activation of complement system and provides an additional useful serum parameter of active SLE.
Oligoclonal T cell expansions in patients with Behçet's disease
Authors: DIRESKENELI H , EKSIOGLU-DEMIRALP E , KIBAROGLU A , YAVUZ S , ERGUN T , AKOGLU T 
Published: March 2003

SUMMARYBehçet's disease (BD) is a multisystem disorder with oral and genital ulcers, mucocutaneous, ocular, joint, vascular and central nervous system involvement. In this study, the peripheral T cell repertoire was analysed in patients with BD with MoAbs against T cell receptor (TCR) Vβ gene products in CD4+ and CD8+ T cell compartments, and these were compared with rheumatoid arthritis (RA) patients and healthy controls (HC). In the CD4+ T cell compartment, oligoclonal TCR Vβ expression was observed in 56% of BD (10/18), 71% of RA (5/7) patients and 21% (3/14) of HC. In the CD8+ T cell group 50% of BD (9/18), 57% of RA patients and 28% of HC (4/14) had an oligoclonal TCR repertoire. An increase of TCR Vβ5.1 subset was observed in five BD patients among CD8+ T cells. Other elevations of TCR Vβ subsets were heterogeneously distributed with one to three different Vβ subsets. Our results suggest an antigen-driven oligoclonal increase of T cells in BD. There was no overall increase in any Vβ group to suggest a superantigen effect. Analysis of the responsible antigens causing the increase in T cell subsets may give insights into the aetiopathogenesis of BD and immunomodulation of these T cells may lead to new treatments.
Prevention of B220+ T cell expansion and prolongation of lifespan induced byLactobacillus caseiin MRL/lpr mice
Authors: Mike A , Nagaoka N , Tagami Y , Miyashita M , Shimada S , Uchida K , Nanno M , Ohwaki M 
Published: March 2003

SUMMARYWe examined the therapeutic effect of heat-killed Lactobacillus casei (LC) on MRL/lpr mice. Ingestion of a diet containing 0.05% (w/w) LC from the weaning period prolonged the lifespan and tended to reduce the proportion of B220+ T cells in the spleen and mesenteric lymph nodes (MLN) of MRL/lpr mice. When LC was intraperitoneally injected once a week after the age of 8 weeks, I-A− macrophages accumulated in the spleen as well as the peritoneum and macrophage progenitors increased in the bone marrow. Moreover, the amount of IL-6 mRNA in peritoneal macrophages was reduced by LC injection. Splenocytes from LC-injected MRL/lpr mice exhibited lower proliferative responses to mitogens than those from control MRL/lpr mice and the increase in number of B220+ T cells in the spleen and MLN was prevented by LC injection. However, LC injection affected neither expression of interferon-gamma (IFN-γ) and IL-4 mRNAs nor proliferative capacities of splenic T cells. Our findings demonstrate that LC injection accelerates macrophage recruitment and prevents the expansion of B220+ T cells without affecting the functions of T cells in MRL/lpr mice. These immunological modulations induced by LC may lead to prolongation of the lifespan of MRL/lpr mice.
Increased levelsin vivoof mRNAs for IL-8 and macrophage inflammatory protein-1α (MIP-1α), but not of RANTES mRNA in peripheral blood mononuclear cells of patients with atopic dermatitis (AD)
Authors: Hatano Y , Katagiri K , Takayasu S 
Published: March 2003

SUMMARYChemokines play an important role in the selective movement of leucocytes into inflammatory areas and they also activate various cells in inflamed tissues. However, it is unclear which cells are the main sources of chemokines in actual inflammatory diseases, even though both mononuclear cells and non-inflammatory resident cells are able to produce chemokines in vitro and the former cells are also the main target of chemokines. To clarify the roles of chemokines that are produced by mononuclear cells in AD, we measured levels in vivo of mRNA for IL-8 and MIP-1α, as well as the level of regulated upon activation normal T cell expressed and secreted (RANTES) mRNA in freshly isolated peripheral blood mononuclear cells from patients with AD. We compared the results with those from psoriatic patients, and patients without AD who were suffering from other cutaneous diseases and eosinophilia. Levels of mRNAs were determined by semiquantitative reverse transcriptase-polymerase chain reactions. Levels of IL-8 and MIP-1α mRNA were elevated not only in atopic patients but also in non-atopic patients with inflammatory skin disease associated with eosinophilia, compared with levels in psoriatic patients and healthy controls. Levels of RANTES mRNA were similar in atopic patients but they were lower in the other two groups of patients when compared with levels in healthy controls. In atopic patients, the levels of both IL-8 and MIP-1α mRNAs but not of RANTES mRNA decreased with improvements in symptom scores after therapy. These findings suggest that mononuclear cells are not only the target of chemokines but might also play an important role in the pathogenesis of AD by producing IL-8 and MIP-1α.
Interferon-alpha (IFN-α) enhances cytotoxicity in healthy volunteers and chronic hepatitis C infection mainly by the perforin pathway
Authors: Kaser A , Enrich B , Ludwiczek O , Vogel W , Tilg H 
Published: March 2003

SUMMARYCell-mediated cytotoxicity is exerted via perforin and Fas ligand (FasL). We have recently shown that IFN-α up-regulates FasL expression in T cells isolated from healthy volunteers and augments activation-induced T cell death. Since the Fas/FasL system is implicated in the pathogenesis of hepatic failure and both molecules have been shown to be up-regulated in hepatitis C virus (HCV) infection, we studied whether cytotoxicity via the FasL system is enhanced by IFN-α and therefore could contribute to hepatic injury. We investigated FasL and perforin expression in peripheral blood mononuclear cells (PBMC) derived from HCV+ donors by Northern analysis and soluble FasL synthesis by ELISA. Natural killer (NK) cell and cytotoxic T lymphocyte (CTL) cytotoxicity was studied by 51Cr-release assays. IFN-α up-regulates FasL mRNA and protein synthesis in mitogen-activated PBMC of HCV+ individuals, as previously found in healthy subjects. Stimulation with IFN-α increases perforin mRNA levels in PBMC. In NK cytotoxicity assays, the enhancement of cytotoxicity by IFN-α is mainly due to the perforin pathway, while the FasL pathway plays only a minor role. In CTL cytotoxicity experiments neither the FasL nor the perforin pathway is further enhanced by IFN-α. Our data suggest that up-regulation of perforin by IFN-α results in elevated cytotoxicity, suggesting that IFN-α might support elimination of virally infected cells via this pathway. In contrast, the major effect of IFN-α on the Fas/FasL system might be the enhanced elimination of activated T cells as a means of finally limiting a T cell response.
Characterization of immune function and analysis of RAG gene mutations in Omenn syndrome and related disorders
Authors: Wada T , Takei K , Kudo M , Shimura S , Kasahara Y , Koizumi S , Kawa-Ha K , Ishida Y , Imashuku S , Seki H , Yachie A 
Published: March 2003

SUMMARY
Omenn syndrome was recently found to be caused by missense mutations in RAG1 or RAG2 gene that result in partial V(D)J recombination activity. Although the clinical hallmarks of the disease are well defined, there have been several cases with clinical findings similar to, but distinct from Omenn syndrome. The data on immune functions and RAG gene mutations of such cases are limited. We described five Japanese infants from four unrelated families, including two cases of Omenn syndrome and three cases of related disorders. Sibling cases with typical Omenn phenotype were found to be compound heterozygotes of R396C and L885R mutations in RAG1. The former has been reported in European cases and may constitute a hot spot. The latter is a novel missense mutation. Infants with related disorders exhibited erythroderma, eosinophilia, hypogammaglobulinaemia, decreased number of B cells and skewing to Th2, and their lymph node specimens showed architectural effacement, lymphocyte depletion and histiocytic hyperplasia, each of which is seen characteristically in Omenn syndrome. However, in these cases serum IgE levels were low or undetectable. We found no mutation in RAG genes except for a K820R substitution in RAG1, which was regarded to be a functional polymorphism, in two of these cases. Our study suggests that RAG missense mutation may be a genetic abnormality unique to Omenn syndrome with characteristic clinical and laboratory findings. Variations of Omenn syndrome, or related disorders, may represent a different type of immunodeficiency, distinct from abnormalities in lymphoid-specific recombinase activity.
Targeted Fc2′-3-PE40 chimeric protein abolishes passive cutaneous anaphylaxis in mice
Authors: Fishman A , Prus D , Belostotsky R , Lorberboum-Galski H 
Published: March 2003

SUMMARYThe alarming increase in the incidence of allergic diseases in the past decade has led to a clear call for more effective treatment. Recently, we reported on the construction of a chimeric protein for targeted elimination of cells expressing FcεRI receptors. This chimeric protein, designated Fc2′-3-PE40, is composed of a Fc fragment of mouse IgE attached to a truncated form of Pseudomonas exotoxin. The Fc2′-3-PE40 chimeric protein was found to be highly cytotoxic to mouse mast cell lines and primary mouse mast cells. We now demonstrate that Fc2′-3-PE40 successfully prevents the development of passive cutaneous anaphylaxis reaction (PCA) in mice. Treatment with Fc2′-3-PE40 for 7 days prevented the PCA reaction in mice by 80% compared with that in control mice given only PBS. Fc2′-3-PE40M, the mutated, enzymatically inactive analogue of Fc2′-3-PE40, did not display this activity. Fc2′-3-PE40 was also effective when given as a single dose 16 h before antigen exposure, resulting in complete inhibition of the PCA reaction. Moreover, treatment with Fc2′-3-PE40 did not cause mast cell degranulation, as the serum histamine values of mice treated with Fc2′-3-PE40 were within the range obtained for control, untreated mice. Thus, the Fc2′-3-PE40 chimeric protein offers a novel approach to the treatment of allergic disorders.