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PubMed2002

CD30-governor of memory T cells?

CD30 is well recognized as a marker expressed by a heterogeneous group of lymphomas and in several immune and autoimmune disorders. However, the function of CD30 in theses diseases or in the normal immune response has remained unclear. Studying gene expression patterns induced by stimulating CD30 signals on a large granular lymphoma cell line, YT, with an agonistic anti-CD30 antibody, we found that CD30 signals affected proapoptotic and antiapoptotic genes, regulated cytotoxicity, and controlled molecules regulating T cell traffic. Creating CD30-L deficient mice and studying CD8 CTL activation and memory responses, it was observed that the absence of CD30-L resulted in diminished primary clonal expansion of CD8 cells. In addition the absence of CD30-L abolished clonal contraction after primary expansion and interfered with secondary expansion upon boosting. The studies suggest that CD30 regulates CD8 CTL function and survival during memory responses and is important for clonal contraction.

Animals↗

PubMed1990

Stability of Markers Used for Identification of Two Rhizobium galegae Inoculant Strains after Five Years in the Field.

The stability of identification markers was examined for two Rhizobium galegae inoculant strains after 5 years in the field. The two strains are genetically closely related, but differ in their lipopolysaccharides. Strain HAMBI 540 has lipopolysaccharide of the rough type, whereas that of strain HAMBI 1461 is of the smooth type. The properties that were examined for 10 field isolates of each inoculant type were symbiotic phenotype, phage type, intrinsic antibiotic resistance, maximum growth temperature, lipopolysaccharide and total soluble protein patterns, immunological properties, DNA restriction profiles, and DNA hybridization patterns, which were determined by using nifHDK and recA sequences as probes. Of these properties, all remained stable in soil, with the exception of some variation in intrinsic antibiotic resistance and the acquisition of an extra EcoRI restriction fragment by one of the isolates. Thus, both the rough and the smooth lipopolysaccharide phenotypes persisted equally well in soil.

Journal Article↗

PubMed1982

Spread of a "Pseudomonas-specific" beta-lactamase to plasmids of enterobacteria.

Eleven isolates including Escherichia coli, Salmonella enteritidis, and Shigella sonnei, obtained in Brazil, Hong Kong, Indonesia, Thailand, and the United States, were found to produce beta-lactamase of the PSE-1 type, which was previously considered to be Pseudomonas specific. The enterobacterial strains produced a beta-lactamase with the same isoelectric point, immunological reactions, and substrate profile as those of the prototype PSE-1 enzyme determined by Pseudomonas plasmid RPL11. The producer strains were resistant to multiple antibiotics, and all contained plasmids, ranging in size from 37 x 10(6) to 130 x 10(6), that belonged to at last six incompatibility groups. Plasmids of IncH2 and IncFIme were shown to contain 8 x 10(6)-molecular-weight transposons Tn1401 and Tn1402 that encoded PSE-1 beta-lactamase production, resistance to streptomycin and spectinomycin via AAD(3"), and resistance to sulfonamide. PSE-1 beta-lactamase was not Pseudomonas specific and appeared to have spread among plasmids found in enterobacteria by transposition.

DNA Transposable Elements↗

PubMed2003

IFNgamma deficient C57BL/6 (H-2b) mice develop collagen induced arthritis with predominant usage of T cell receptor Vbeta6 and Vbeta8 in arthritic joints.

BACKGROUND: Transgenic deficiency in interferon gamma (IFNgamma) or IFNgamma receptor makes resistant strains of mice bearing H-2(b) or H-2(d) susceptible to collagen induced arthritis (CIA). OBJECTIVE: To determine whether the escape from regulation of disease susceptibility at the major histocompatibility complex level involves a new use of autoimmune T cells expressing T cell receptor (TCR) Vbeta that vary from the cell populations previously identified within arthritic joints. METHODS: Arthritis was induced by a standard protocol with type II bovine collagen (CII) in complete Freund's adjuvant. Clinical features, histopathology, immunological responses, and TCR profile in arthritic joints in IFNgamma knockout C57BL/6 (B6.IFNgamma KO) mice (H-2(b)) were compared directly with those in DBA/1 mice (H-2(q)). RESULTS: 60-80% of B6.IFNgamma KO mice developed a progressive arthritis with a similar clinical course to classical CIA in DBA/1 mice. The affected joints in B6.IFNgamma KO mice had an erosive form of arthritis with similar features to joint disease in DBA/1 mice. B6.IFNgamma KO mice produced significantly higher levels of IgG2b and IgG1 autoantibodies to murine CII and showed increased proliferative response to CII compared with B6 mice. Comparable levels of interleukin 1beta and tumour necrosis factor alpha expression were detected in arthritic joints from beta6.IFNgamma KO and DBA/1 mice. B6.IFNgammaKO mice used predominantly TCR Vbeta6 and Vbeta8 in arthritic joints. This TCR Vbeta profile is similar to that found in DBA/1 mice with CIA. CONCLUSIONS: C57BL/6 mice deficient in IFNgamma production can develop arthritis that resembles classical CIA. These data suggest that IFNgamma is a key factor mediating susceptibility to CIA.

Animals↗

PubMed1994

Biochemical evidence of a kallikrein-like activity in rat reproductive tissues.

We performed this study to examine the presence of a kallikrein-kinin system in rat fetal and maternal tissues. Uteri and placenta from Wistar pregnant and nonpregnant rats were perfused to eliminate blood, and fetal membranes were washed several times with saline. Amniotic fluids were obtained without blood contamination by amniocentesis from eight rats. The different samples were homogenized and centrifuged (2000g during 20 minutes), and the supernatant was incubated with dog kininogen and 0.1 mol/L Tris-HCl buffer (pH 8.5) in the presence of peptidase inhibitors. Kinins released were measured by radioimmunoassay. Kininogenase activity was found in rat uteri, placental vessels, amniotic fluids, and fetal membranes. The enzymes were present in active but mostly in inactive forms. The kallikrein-like enzymes found in the different preparations and rat urinary kallikrein used as control had similar molecular weights, immunologic characteristics, and inhibition profiles with protease inhibitors. We conclude that kallikrein-like enzymes are present in rat organs of reproduction. These data suggest that kinins released locally may act as paracrine hormones in the regulation of blood pressure during pregnancy.

Amniotic Fluid↗

PubMed2007

Cartography of gene expression in CD8 single cells: novel CCR7- subsets suggest differentiation independent of CD45RA expression.

Understanding the distribution, function, and lineage relationship of CD8+ T-cell subpopulations is of fundamental value for the monitoring of the immune system in several experimental and clinical situations. However, the available data concerning the description of effector and memory CD8+ subsets in humans remain rather fragmentary because different studies favored the usage of distinct and restricted sets of cell surface markers and functional parameters. We associated multiple markers to subdivide CD8+ T cells into 14 different cell types, several of which were not described previously, and evaluated the coexpression of 18 genes simultaneously in individual cells from each subset. Our results show that each subset has a defined pattern of gene expression. Moreover, effector gene expression of CCR7- cells correlated only with CD27 expression levels and CD27/CD28 coexpression but not with CD45RA/R0 phenotypes. Our findings thus describe new CD8+ cell subsets, allow the identification of relatively homogeneous CD8+ subpopulations, provide a predictable and precise correlation between particular cell surface markers and CD8+ T-cell functional properties, and identify effector cells present in both CCR7-CD45RA+ and CCR7-CD45R0+ compartments. The results also indicate that activated cells might modulate the expression of CD45RA/R0 asynchronously rather than CCR7-CD45RA+ cells always issuing from CD45RA- precursors.

Adult↗

PubMed2004

Differential gene expression by integrin beta 7+ and beta 7- memory T helper cells.

BACKGROUND: The cell adhesion molecule integrin alpha 4 beta 7 helps direct the migration of blood lymphocytes to the intestine and associated lymphoid tissues. We hypothesized that beta 7+ and beta 7- blood memory T helper cells differ in their expression of genes that play a role in the adhesion or migration of T cells. RESULTS: RNA was prepared from beta 7+ and beta 7- CD4+ CD45RA- blood T cells from nine normal human subjects and analyzed using oligonucleotide microarrays. Of 21357 genes represented on the arrays, 16 were more highly expressed in beta 7+ cells and 18 were more highly expressed in beta 7- cells (>/=1.5 fold difference and adjusted P < 0.05). Several of the differentially expressed transcripts encode proteins with established or putative roles in lymphocyte adhesion and chemotaxis, including the chemokine receptors CCR9 and CCR10, the integrin alpha 4 subunit, L-selectin, KLRB1 (CD161), NT5E (CD73), LGALS1 and LGALS2 (galectin-1 and -2), and RGS1. Flow cytometry was used to determine whether differences in levels of transcripts encoding cell surface proteins were associated with differential expression of those proteins. Using this approach, we found that surface expression of KLRB1, LAIR1, and NT5E proteins was higher on beta 7+ memory/effector T cells than on beta 7- cells. CONCLUSIONS: Memory/effector T cells that express integrin beta 7 have a distinct pattern of expression of a set of gene transcripts. Several of these molecules can affect cell adhesion or chemotaxis and are therefore likely to modulate the complex multistep process that regulates trafficking of CD4+ memory T cell subsets with different homing behaviors.

Adult↗

PubMed2005

Plasmodium yoelii 17XL infection up-regulates RANTES, CCR1, CCR3 and CCR5 expression, and induces ultrastructural changes in the cerebellum.

BACKGROUND: Malaria afflicts 300-500 million people causing over 1 million deaths globally per year. The immunopathogenesis of malaria is mediated partly by co mplex cellular and immunomodulator interactions involving co-regulators such as cytokines and adhesion molecules. However, the role of chemokines and their receptors in malaria immunopathology remains unclear. RANTES (Regulated on Activation Normal T-Cell Expressed and Secreted) is a chemokine involved in the generation of inflammatory infiltrates. Recent studies indicate that the degradation of cell-cell junctions, blood-brain barrier dysfunction, recruitment of leukocytes and Plasmodium-infected erythrocytes into and occlusion of microvessels relevant to malaria pathogenesis are associated with RANTES expression. Additionally, activated lymphocytes, platelets and endothelial cells release large quantities of RANTES, thus suggesting a unique role for RANTES in the generation and maintenance of the malaria-induced inflammatory response. The hypothesis of this study is that RANTES and its corresponding receptors (CCR1, CCR3 and CCR5) modulate malaria immunopathogenesis. A murine malaria model was utilized to evaluate the role of this chemokine and its receptors in malaria. METHODS: The alterations in immunomodulator gene expression in brains of Plasmodium yoelii 17XL-infected mice was analysed using cDNA microarray screening, followed by a temporal comparison of mRNA and protein expression of RANTES and its corresponding receptors by qRT-PCR and Western blot analysis, respectively. Plasma RANTES levels was determined by ELISA and ultrastructural studies of brain sections from infected and uninfected mice was conducted. RESULTS: RANTES (p < 0.002), CCR1 (p < 0.036), CCR3 (p < 0.033), and CCR5 (p < 0.026) mRNA were significantly upregulated at peak parasitaemia and remained high thereafter in the experimental mouse model. RANTES protein in the brain of infected mice was upregulated (p < 0.034) compared with controls. RANTES plasma levels were significantly upregulated; two to three fold in infected mice compared with controls (p < 0.026). Some distal microvascular endothelium in infected cerebellum appeared degraded, but remained intact in controls. CONCLUSION: The upregulation of RANTES, CCR1, CCR3, and CCR5 mRNA, and RANTES protein mediate inflammation and cellular degradation in the cerebellum during P. yoelii 17XL malaria.

Animals↗

PubMed2026

Transcriptome-based high-frequency recurrence index predicts frequent recurrence in non-muscle-invasive bladder cancer after Bacillus Calmette-Guérin therapy.

BACKGROUND: High-frequency recurrence (HfR, ≥ 2 recurrences) in non-muscle-invasive bladder cancer (NMIBC) poses a significant clinical burden. Current risk models, such as the European Organization for Research and Treatment of Cancer (EORTC), the European Association of Urology (EAU), and the UROMOL classification, offer limited predictive accuracy for identifying patients at risk for frequent recurrence despite appropriate treatment. METHODS: A 75-gene high-frequency recurrence index (HfRI) was constructed by selecting recurrence-associated genes using differential expression and Cox regression analyses. The HfRI was computed as a weighted sum of normalized gene expression values. The model was trained on a discovery cohort and validated in multiple cohorts (n = 1379) using machine-learning approaches. Clinical relevance was assessed using recurrence-free survival (RFS) and Cox models, and predictive performance was compared with that of the EORTC, EAU, and UROMOL classifications using the area under the curve (AUC) and the concordance index (c-index). RESULTS: The HfRI robustly stratified patients into high-risk and low-risk groups across six independent NMIBC cohorts. Patients classified as HfRI-high had a significantly greater likelihood of experiencing ≥ 2 recurrences (χ2, p = 0.001) and showed markedly reduced RFS (log-rank test, p < 0.001). The adverse prognostic effect of the HfRI persisted even among patients treated with BCG therapy (log-rank test, p = 0.02). Multivariate analysis revealed that the HfRI was an independent predictor of HfR (HR = 2.82, 95% CI = 1.89-4.20, p < 0.001). Compared with established clinical risk classifiers, the HfRI demonstrated superior predictive performance (AUC = 0.736, c-index = 0.673) in terms of the EORTC (AUC = 0.594), EAU (AUC = 0.557) risk groups, and UROMOL2021 (AUC = 0.596) classification. Pathway analysis revealed that HfRI-high tumors were characterized by upregulation of cell cycle progression and DNA replication pathways, accompanied by suppression of immune signaling pathways. These biological features provide a mechanistic explanation for the reduced responsiveness to intravesical BCG therapy, underscoring the role of HfRI not only as a predictor of recurrence risk but also as a biomarker capable of identifying patients unlikely to benefit from standard BCG treatment. CONCLUSIONS: HfRI represents a robust, transcriptome-based tool for predicting frequent recurrence in NMIBC patients. The HfRI supports earlier identification of patients at risk of high-frequency recurrence, thereby supporting personalized treatment strategies.

Humans↗

PubMed1983

Effect of N-(2-carboxyphenyl)-4-chloroanthranilic acid disodium salt (CCA) on the induction of helper and suppressor T cells in vitro and in vivo.

Keyhole lympet hemocyanin (KLH)-specific suppressor T (Ts) cells that suppress the in vitro secondary anti-trinitrophenyl (TNP) PFC response to TNP-KLH could be induced when murine spleen cells were precultured with KLH. N-(2-carboxyphenyl)-4-chloroanthranilic acid disodium salt (CCA) at 1-100 micrograms/ml augmented the in vitro induction of Ts cells when the cells were precultured with a suboptimal dose of KLH (10 micrograms/ml). Ts cell-induction was, however, rather slightly inhibited by the same concentrations of CCA when the lymphocytes were precultured with an optimal amount of KLH (100 micrograms/ml). In the in vivo experiments, the daily administration of 10 mg/kg CCA for 4 weeks augmented or inhibited Ts cell-induction when mice were immunized with a suboptimal (30 micrograms/body) or an optimal (100 micrograms/body) amount of KLH, respectively. However, CCA had no effect on the induction of Ts cells by concanavalin A in vivo. On the other hand, CCA augmented the induction of helper T (Th) cells both in vitro and in vivo when Th cells were induced with a suboptimal amount of antigens. In contrast, the augmentative effect was no longer observed when Th cells were induced by an optimal amount of antigens. These results suggest that CCA is a compound showing immunomodulating properties that affect Ts and Th cell-induction depending on immunological conditions. These immunopharmacological profiles are discussed in connection with its clinical application to an autoimmune disease like rheumatoid arthritis.

Animals↗

PubMed1993

Biological and immunological properties of the international standard for FSH 83/575: isoelectrofocusing profile and comparison with other FSH preparations.

The new international standard for FSH, IS 83/575, has been analyzed, after isoelectric focusing separation, by Sertoli cell in vitro bioassay, radioligand receptor assay and two highly specific immunometric assays. Its molecular composition was then compared with the isoelectric focusing profiles obtained from the fractionation of the reference preparation 2nd IRP 78/549 and from pools of human male and female pituitary extracts and male and female sera. The results showed that > 80% of immunoreactive and bioactive FSH in the IS 83/575 has a pI value < 4, while such very acidic material was represented much less in the other FSH preparations tested. All the immunoreactive material contained in the IS 83/575 was shown to be capable of receptor binding and bioactivity in vitro. A generally good correspondence between IEF profiles obtained by bioassay and by immunofluorimetric assay was evident in the case of IS 83/575, 2nd IRP 78/549 and pituitary extracts, although the profiles recorded by immunofluorimetric assay were rather smooth and more isoforms were detected by bioassay. A striking discrepancy between immunoreactive FSH and bioactive FSH was observed after isoelectric focusing fractionation of the serum pools, in which some bioactive material was not detected by immunofluorimetric assay and some of the immunoreactive FSH peaks were devoid of bioactivity, indicating that serum contains inhibitors of FSH action and that immunometric assays based on monoclonal antibodies may miss some bioactive FSH isoforms.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

PubMed2006

A behavioral and cognitive profile of clinically stable HIV-infected children.

OBJECTIVE: The purpose of this research was to characterize behavioral and cognitive profiles of clinically and immunologically stable antiretroviral-experienced HIV-infected children. METHODS: Two hundred seventy-four previously treated HIV-infected children aged 2 to 17 years were assessed for behavioral, developmental, and cognitive functioning. Correlations between neuropsychological measures, age, and CD4 lymphocyte count were investigated. RESULTS: The most common behavioral problems, as measured by the Conners' Parent Rating Scale, were psychosomatic (28%), learning (25%), hyperactivity (20%), impulsive-hyperactive (19%), conduct (16%), and anxiety (8%) problems. Mean Wechsler Intelligence Scale for Children-III scores were less than established population norms; the mean verbal IQ was 85, the mean performance IQ was 90, and the mean full-scale score was 86. Hyperactivity was more frequent in children with a Wechsler Intelligence Scale for Children-III performance IQ of <90. Anxiety problems were more likely in children > or =9 years of age. Children with CD4 counts of <660 cells per mm3 were more likely to be identified as having a conduct disorder. No association was noted between behavioral problems and neuroimaging. CONCLUSIONS: Clinically and immunologically stable HIV-infected children had more frequent behavioral problems and lower developmental and cognitive scores than established childhood norms.

Adolescent↗

PubMed2005

Molecular signatures distinguish human central memory from effector memory CD8 T cell subsets.

Memory T cells are heterogeneous in terms of their phenotype and functional properties. We investigated the molecular profiles of human CD8 naive central memory (T(CM)), effector memory (T(EM)), and effector memory RA (T(EMRA)) T cells using gene expression microarrays and phospho-protein-specific intracellular flow cytometry. We demonstrate that T(CM) have a gene expression and cytokine signaling signature that lies between that of naive and T(EM) or T(EMRA) cells, whereas T(EM) and T(EMRA) are closely related. Our data define the molecular basis for the different functional properties of central and effector memory subsets. We show that T(EM) and T(EMRA) cells strongly express genes with known importance in CD8 T cell effector function. In contrast, T(CM) are characterized by high basal and cytokine-induced STAT5 phosphorylation, reflecting their capacity for self-renewal. Altogether, our results distinguish T(CM) and T(EM)/T(EMRA) at the molecular level and are consistent with the concept that T(CM) represent memory stem cells.

CD8-Positive T-Lymphocytes↗

PubMed2001

Cytokine gene expression in neoplastic B cells from human mantle cell, follicular, and marginal zone lymphomas and in their postulated normal counterparts.

Cytokines may promote tumor growth by paracrine and/or autocrine pathways. Little information is available because malignant cells differ from their normal counterparts for the cytokine repertoire they express. Here we have investigated by reverse transcription-PCR the expression of 22 cytokine genes in neoplastic B lymphocytes from six patients with mantle cell lymphoma, 10 with follicular lymphoma, and 5 with marginal zone lymphoma and in their normal counterparts, i.e., naive, germinal center, and memory B cells, purified from tonsils. The overall profiles of cytokine gene expression in neoplastic B cells and in the corresponding normal B-cell subsets were similar, but some "holes" in the repertoire of malignant versus normal B lymphocytes were detected. Different "hole" combinations were identified consistently in mantle cell lymphoma, follicular lymphoma, and marginal zone lymphoma, thus representing molecular fingerprints of each individual lymphoma entity.

B-Lymphocytes↗

PubMedDate not supplied

Gene expression profiling of peripheral blood mononuclear leukocytes from psoriasis patients identifies new immune regulatory molecules.

In the present report gene expression profiling of peripheral blood mononuclear cells (PBMC) from psoriasis patients suffering from severe generalized disease was performed comparing diseased stage with cured stage. By this means, 18 genes were identified which showed differential expression. The most significant differences were found for IL-8, annexin A3, cycloxygenase-2 (COX-2), cell cycle regulator G0S2, and pre-B cell enhancing factor (PBEF), all of which showed upregulation in the diseased stage. Microarray data were confirmed by real-time RT-PCR. Further analyses using support vector machines identified three pairs of genes (IL-8 - CDKN1C/p57, cyclooxygenase-2 - NR1D2, and desmocollin-2 - CDKN1C/p57) which allowed an accuracy of disease stage prediction of 86%, based on gene expression patterns. Taken together, this is the first large-scale gene expression study of psoriasis PBMC identifying candidate genes that might contribute to psoriasis immunopathogenesis. The genes identified in the present report and the molecular mechanisms underlying their regulation might serve as future targets for therapeutic intervention in psoriasis.

Gene Expression Profiling↗

PubMed2005

Recent advances in gene expression data clustering: a case study with comparative results.

Several advanced techniques have been proposed for data clustering and many of them have been applied to gene expression data, with partial success. The high dimensionality and the multitude of admissible perspectives for data analysis of gene expression require additional computational resources, such as hierarchical structures and dynamic allocation of resources. We present an immune-inspired hierarchical clustering device, called hierarchical artificial immune network (HaiNet), especially devoted to the analysis of gene expression data. This technique was applied to a newly generated data set, involving maize plants exposed to different aluminum concentrations. The performance of the algorithm was compared with that of a self-organizing map, which is commonly adopted to deal with gene expression data sets. More consistent and informative results were obtained with HaiNet.

Algorithms↗

PubMed1989

[Experimental studies on the stimulation of prenatal development in swine by PMSG treatment in early pregnancy. 2. Analytic studies of hormones in young sows after PMSG treatment on the 11th day of pregnancy].

Tens gilts received 400 I.U. of Pregmagon on the 11th day of pregnancy (day of KB2 = 1 day of pregnancy), while 8 sows were used as controls. Blood was sampled 4 times a day between the 11th and 25th day of pregnancy, with an indwelling catheter being used. Progesterone, oestradiol-17 beta, and luteinising hormone were radio-immunologically determined. The hormone profiles of pregnant sows were found to differ unambiguously from those of non-pregnant animals for both concentrations and frequencies. They were indicative of close correlations and time links of gonadotrophic and steroid hormones in early pregnancy and in oestrus. PMSG treatment in pregnancy resulted in increase in the oestradiol-17 beta concentration and attenuation in pulsatile luteinising hormone release at the time of implantation. Progesterone concentrations in peripheral blood were not affected by PMSG injection.

Animals↗

PubMed1995

[Laboratory medicine of lymphocytes].

Recently, many laboratory tests associated with lymphocytes and/or lymphocyte functions have been developed. Among them, cytokine determinations of various body fluids and cytokine production assays are most highlightened. As cytokine is an effector molecule in immunological network, cytokine production profile may indicate functional state of immune response and immunopathological process. On the basis of these understandings, we propose cytokine production assay of whole blood culture system.

Cytokines↗