bluuebunny/crossref_metadata_2025 huggingface dataset:
Microsatellite diversity, population subdivision and gene flow in the Lipizzan horse
Authors: Achmann R. , Curik I. , Dovc P. , Kavar T. , Bodo I. , Habe F. , Marti E. , Sölkner J. , Brem G.
Published: July 2004

Blood samples of 561 Lipizzan horses from subpopulations (studs) of seven European countries representing a large fraction of the breed's population were used to examine the genetic diversity, population subdivision and gene flow in the breed. DNA analysis based on 18 microsatellite loci revealed that genetic diversity (observed heterozygosity = 0.663, gene diversity = 0.675 and the mean number of alleles = 7.056) in the Lipizzan horse is similar to other horse breeds as well as to other domestic animal species. The genetic differentiation between Lipizzan horses from different studs, although moderate, was apparent (pairwise FSTcoefficients ranged from 0.021 to 0.080). Complementary findings explaining the genetic relationship among studs were revealed by genetic distance and principal component analysis. One genetic cluster consisted of the subpopulations of Austria, Italy and Slovenia, which represent the classical pool of Lipizzan horse breeding. A second cluster was formed by the Croatian, Hungarian and Slovakian subpopulations. The Romanian subpopulation formed a separate unit. The largest genetic differentiation was found between the Romanian and Italian subpopulation. Genetic results are consistent with the known breeding history of the Lipizzan horse. Correct stud assignment was obtained for 80.9% and 92.1% of Lipizzan horses depending on the inclusion or exclusion of migrant horses, respectively. The results of the present study will be useful for the development of breeding strategies, which consider classical horse breeding as well as recent achievements of population and conservation genetics.
B‐F DNA sequence variability in Brazilian (blue‐egg Caipira) chickens
Authors: Lima‐Rosa C. A. V. , Canal C. W. , Streck A. F. , Freitas L. B. , Delgado‐Cañedo A. , Bonatto S. L. , Salzano F. M.
Published: July 2004

A total of 100 chickens from the Brazilian (blue‐egg Caipira) native breed were studied in relation to exon 2 of the B‐F genes of the major histocompatibility complex (MHC) region. After a first screening on 100 birds, 22 animals were selected for amplification, cloning and sequencing experiments of exons 2–4 (a total of 1048 bp) of their DNA. Twenty‐three sequences were obtained, of which at least 10 appear novel. Inferred protein sequences were compared with those previously described, totalling 41 different sequences with amino acid changes in 33 of the 88 sites in α1, and 34 of the 91 sites in α2 domains. Allele expression was investigated in these animals through cloning experiments. The blue‐egg Caipira chickens may provide a source of novel B‐F alleles for genetic improvement programmes.
Comparative mapping of human chromosome 10 to pig chromosomes 10 and 14*
Authors: Nonneman D. , Rohrer G. A.
Published: July 2004

Identification of predictive markers in QTL regions that impact production traits in commercial populations of swine is dependent on construction of dense comparative maps with human and mouse genomes. Chromosomal painting in swine suggests that large genomic blocks are conserved between pig and human, while mapping of individual genes reveals that gene order can be quite divergent. High‐resolution comparative maps in regions affecting traits of interest are necessary for selection of positional candidate genes to evaluate nucleotide variation causing phenotypic differences. The objective of this study was to construct an ordered comparative map of human chromosome 10 and pig chromosomes 10 and 14. As a large portion of both pig chromosomes are represented by HSA10, genes at regularly spaced intervals along this chromosome were targeted for placement in the porcine genome. A total of 29 genes from human chromosome 10 were mapped to porcine chromosomes 10 (SSC10) and 14 (SSC14) averaging about 5 Mb distance of human DNA per marker. Eighteen genes were assigned by linkage in the MARC mapping population, five genes were physically assigned with the IMpRH mapping panel and seven genes were assigned on both maps. Seventeen genes from human 10p mapped to SSC10, and 12 genes from human 10q mapped to SSC14. Comparative maps of mammalian species indicate that chromosomal segments are conserved across several species and represent syntenic blocks with distinct breakpoints. Development of comparative maps containing several species should reveal conserved syntenic blocks that will allow us to better define QTL regions in livestock.
Genetic diversity in German draught horse breeds compared with a group of primitive, riding and wild horses by means of microsatellite DNA markers
Authors: Aberle K. S. , Hamann H. , Drögemüller C. , Distl O.
Published: July 2004

We compared the genetic diversity and distance among six German draught horse breeds to wild (Przewalski's Horse), primitive (Icelandic Horse, Sorraia Horse, Exmoor Pony) or riding horse breeds (Hanoverian Warmblood, Arabian) by means of genotypic information from 30 microsatellite loci. The draught horse breeds included the South German Coldblood, Rhenish German Draught Horse, Mecklenburg Coldblood, Saxon Thuringa Coldblood, Black Forest Horse and Schleswig Draught Horse. Despite large differences in population sizes, the average observed heterozygosity (Ho) differed little among the heavy horse breeds (0.64–0.71), but was considerably lower than in the Hanoverian Warmblood or Icelandic Horse population. The mean number of alleles (NA) decreased more markedly with declining population sizes of German draught horse breeds (5.2–6.3) but did not reach the values of Hanoverian Warmblood (NA = 6.7). The coefficient of differentiation among the heavy horse breeds showed 11.6% of the diversity between the heavy horse breeds, as opposed to 21.2% between the other horse populations. The differentiation test revealed highly significant genetic differences among all draught horse breeds except the Mecklenburg and Saxon Thuringa Coldbloods. The Schleswig Draught Horse was the most distinct draught horse breed. In conclusion, the study demonstrated a clear distinction among the German draught horse breeds and even among breeds with a very short history of divergence like Rhenish German Draught Horse and its East German subpopulations Mecklenburg and Saxon Thuringa Coldblood.
Association of the heart fatty acid‐binding protein (FABP3) gene with milk traits in Manchega breed sheep
Authors: Calvo J. H. , Marcos S. , Jurado J. J. , Serrano M.
Published: July 2004

The ovine fatty acid‐binding protein type 3 gene has been chosen as a functional candidate gene for milk traits. Two different single nucleotide polymorphisms (SNPs) of ovine FABP3 gene have been tested in a daughter design comprising 13 families. No association was found between estimated breeding values for milk yield, protein and fat contents (FC) and genotypes across families using anova and transmission disequilibrium test (TDT). In within‐family analysis, one family showed a significant effect for FC. These results could indicate linkage disequilibrium between the FABP3 gene and a quantitative trait loci (QTL) for FC, with the heterozygous genotype associated with a positive effect in this trait.
HTLV-1 modifies the clinical and immunological response to schistosomiasis
Authors: PORTO A F , SANTOS S B , ALCÂNTARA L , GUERREIRO J B , PASSOS J , GONZALEZ T , NEVA F , GONZALEZ D , HO J L , CARVALHO E M
Published: July 2004

The immunological response in HTLV-1 infected individuals is characterized by a prominent Type-1 cytokine response with high production of IFN-γ and TNF-α. In contrast, helminthic infections and in particular chronic schistosomiasis are associated with a predominant production of IL-4, IL-5, IL-10 and IL-13. Liver fibrosis is the main pathological finding in schistosomiasis that occurs after many years of infection. This pathology is T cell dependent but the immune response mechanisms are not completely understood. The North-east region of Brazil is endemic for both HTLV-1 and schistosomiasis. In the present study the immune response, clinical severity, and therapeutic response to praziquantel of patients with schistosomiasis coinfected with HTLV-1 were compared with patients infected only with S. mansoni. Patients with HTLV-1 and S. mansoni had lower levels of IL-5 (P < 0·05) and higher levels of IFN-γ (P < 0·05) in cultures stimulated with S. mansoni antigen and decreased S. mansoni antigen specific IgE levels when compared with patients with schistosomiasis without HTLV-1 coinfection. Liver fibrosis was mild in all HTLV-1 coinfected patients and efficacy of praziquantel was lower in patients dually infected than in patients infected only with S. mansoni.
T cell-derived tumour necrosis factor is essential, but not sufficient, for protection againstMycobacterium tuberculosisinfection
Authors: SAUNDERS B M , BRISCOE H , BRITTON W J
Published: July 2004

Tumour necrosis factor (TNF) is critical for sustained protective immunity against Mycobacterium tuberculosis infection. To investigate the relative contributions of macrophage- and T cell-derived TNF towards  this immunity T cells from wild-type (WT) or TNF–/– mice were transferred into RAG–/– or TNF–/– mice which were then infected with M. tuberculosis. Infected RAG–/– mice and RAG–/– recipients of TNF deficient T cells developed overwhelming infection, with extensive pulmonary and hepatic necrosis and succumbed with a median of only 16 days infection. By contrast, RAG–/– recipients of WT T cells showed a significant increase in survival with a median of 32 days. Although initial bacterial growth was similar in all groups of RAG–/– mice, the transfer of WT, but not TNF–/–, T cells led to the formation of discrete foci of leucocytes and macrophages and delayed the development of necrotizing pathology. To determine requirements for macrophage-derived TNF, WT or TNF–/– T cells were transferred into TNF–/– mice at the time of M. tuberculosis infection. Transfer of WT T cells significantly prolonged survival and reduced the early tissue necrosis evident in the TNF–/– mice, however, these mice eventually succumbed indicating that T cell-derived TNF alone is insufficient to control the infection. Therefore, both T cell- and macrophage-derived TNF play distinct roles in orchestrating the protective inflammatory response and enhancing survival during M. tuberculosis infection.
Reduction of human anti-tetanus toxoid antibody in hu-PBL-SCID mice by immunodominant peptides of tetanus toxoid
Authors: JACKSON D J , ELSON C J , KUMPEL B M
Published: July 2004

Immunotherapy of murine autoimmune and allergic diseases by administration of peptides corresponding to the dominant T cell epitope is a reality. However, problems remain in applying this therapy to reduce antibody responses in humans. To overcome these difficulties, a preclinical system was developed to test the effect of immunodominant peptides from a common antigen, tetanus toxoid (TT), on the long-term human anti-TT response. Individuals whose T cells proliferated against dominant TT peptides were identified. Peripheral blood leucocytes (PBL) from these donors were injected intraperitoneally (i.p.) into mice with severe combined immunodeficiency (SCID) that had been depleted of murine natural killer (NK) cells (hu-PBL-SCID mice). Peptides or PBS were injected i.p. before a further injection of PBL and immunization with TT. The concentration of human IgG and anti-TT in murine plasma was followed for 10 weeks. The total IgG was similar in both groups. By contrast, there was a statistically significant reduction in IgG anti-TT from eight weeks onwards. It is considered that the hu-PBL-SCID model system may provide a means by which the efficacy of peptide immunotherapy for reduction of pathological antibodies in humans can be examined.
CD56+dim and CD56+bright cell activation and apoptosis in hepatitis C virus infection
Authors: LIN A W , GONZALEZ S A , CUNNINGHAM-RUNDLES S , DORANTE G , MARSHALL S , TIGNOR A , HA C , JACOBSON I M , TALAL A H
Published: July 2004

CD3–CD56+dim natural killer (NK) cells, which are cytotoxic against virally infected cells, may be important in hepatitis C virus (HCV)-infected patients who are successfully treated with pegylated interferon (PEG-IFN)-α. We used flow cytometry to enumerate activated (CD69+) and apoptotic (annexin-V+) dim (CD3–CD56+dim) and bright (CD3–CD56+bright) NK cells obtained from HCV-infected patients before treatment (n = 16) and healthy controls (n = 15) in the absence and presence of pegylated interferon (PEG-IFN)-α-2b. A subset of HCV-infected patients, subsequently treated with PEG-IFN-α-2b in vivo, was determined to have a sustained virological response (SVR, n = 6) or to not respond (NR) to treatment (n = 5). In the absence of IFN, activated dim (CD3–CD56+dim CD69+) NK cells were significantly decreased (P = 0·04) while activated apoptotic dim (CD3–CD56+dimCD69+annexin-V+) NK cells tended to be increased (P = 0·07) in SVR patients compared with NR patients. Activated bright (CD3–CD56+brightCD69+) and activated apoptotic bright (CD3–CD56+brightCD69+annexin-V+) NK cells were significantly correlated (P = 0·02 and P = 0·01, respectively) with increasing hepatic inflammation. These findings suggest that in the absence of PEG-IFN, activated dim (CD3–CD56+dimCD69+) NK cell turnover may be enhanced in SVR compared with NR patients and that activated bright (CD3–CD56+brightCD69+) NK cells may play a role in liver inflammation.
Immunomodulation by roquinimex decreases the expression of IL-23 (p19) mRNA in the brains of herpes simplex virus type 1 infected BALB/c mice
Authors: PELTONIEMI J , BROBERG E K , HALENIUS A , SETÄLÄ N , ERÄLINNA J-P , SALMI A A , RÖYTTÄ M , HUKKANEN V
Published: July 2004

Herpes simplex virus (HSV) is a common neurotropic virus which infects epithelial cells and subsequently the trigeminal ganglia (TG) and brain tissue. We studied how immunomodulation with roquinimex (Linomide®) affects the course of corneal HSV infection in BALB/c mice. BALB/c mice have also been used in a model for HSV-based vectors in treating an autoimmune disease of the central nervous system (CNS). We addressed the questions of how immunomodulation affects the local as well as the systemic immune response and whether roquinimex could facilitate the spread of HSV to the CNS. The cytokine response in the brain and TG was studied using a quantitative rapid real-time RT-PCR method. We were interested in whether immunomodulation affects the expression of the recently described Th1-cytokine IL-23p19 in the brain and TG. The expression of IL-23 mRNA was decreased in brains of roquinimex-treated BALB/c mice. Also the expression of IL-12p35 and IFN-γ mRNAs decreased. No significant changes were seen in IL-4 and IL-10 mRNA expression. The cytokine response was also studied using supernatants of stimulated splenocytes by EIA. Roquinimex treatment suppressed the production of IFN-γ and also the production of IL-10 in HSV-infected BALB/c mice.
Nasal mucosa in natural colds: effects of allergic rhinitis and susceptibility to recurrent sinusitis
Authors: ALHO O-P , KARTTUNEN R , KARTTUNEN T J
Published: July 2004

The mechanisms of virus-induced airway hyperresponsiveness in asthma and allergy and the failure of host defence in patients suffering from secondary airway infections are still largely unknown. The aim of this study was to examine whether the presence of allergic rhinitis or susceptibility to recurrent sinusitis affects the structural and cellular changes in nasal mucosa during natural colds and convalescence. We compared the mucosal changes in biopsy samples during acute natural colds (days 2–4 of illness) and convalescence (3 weeks later) in patients with allergic rhinitis (n = 9), patients with susceptibility to sinusitis (n = 19) and healthy controls (n = 20). We saw similarly increased numbers of mucosal T and B lymphocytes and mast cells and increased vascular density during the acute colds compared to convalescence in all the three groups. The allergic subjects had elevated levels of eosinophils in the acute phase (P = 0·03), and the allergic and sinusitis-prone subjects had elevated levels of epithelial T cells (P = 0·04) and low levels of mast cells (P = 0·005) in convalescence compared to the control group. The sinusitis-prone subjects lacked intraepithelial cytotoxic cells in convalescence. In the allergic subjects, the reticular basement membrane was thicker in the acute phase compared to the convalescence (P = 0·05). These results suggest that various cells of the airways, including inflammatory and structural cells, are involved during viral respiratory infections in subjects with allergic rhinitis. The small numbers of mast cells and cytotoxic lymphocytes in the sinusitis-prone subjects may be related to their susceptibility to bacterial complications.
Environmental factors and not genotype influence the plasma level of interleukin-1 receptor antagonist in normal individuals
Authors: CULLUP H , MIDDLETON P G , DUGGAN G , CONN J S , DICKINSON A M
Published: July 2004

Cytokine production may be regulated by both genotypic (single nucleotide or tandem repeat polymorphisms) and non-genotypic factors relating to the environment and inherent biology (i.e. gender). Interleukin (IL)-1 is one of the body's most highly proinflammatory cytokines and is implicated in the pathophysiology of numerous diseases, but also in the maintenance of homeostasis in a number of tissues. The cytokine IL-1 receptor antagonist (IL-1Ra) is the competitive inhibitor of the IL-1 agonists IL-1α and IL-1β. In vivo IL-1Ra was measured in a cohort of 200 + blood donors and the effect of the IL-1 gene polymorphisms, environmental and biological factors assessed. In this study, we observed that possession of particular alleles of 5 IL-1 gene polymorphisms (IL1A-889, IL1Α VNTR, IL1B -511, IL1B +3953 and the IL1RN VNTR) did not correlate with higher plasma IL-1Ra levels. Environmental factors such as smoking and non-steroidal anti-inflammatory drug ingestion were associated with higher in vivo IL-1Ra levels (P = 0·015 and 0·022, respectively), but biological factors such as gender, age and menstruation status did not have any impact upon in vivo IL-1Ra levels. Genotypic associations of IL-1 gene family polymorphisms with disease features may reflect characteristics of stressed rather than normal control circuits for cytokine production.
Beneficial effects of troglitazone on neutrophil dysfunction in multiple low-dose streptozotocin-induced diabetic mice
Authors: KANNAN Y , TOKUNAGA M , MORIYAMA M , KINOSHITA H , NAKAMURA Y
Published: July 2004

Patients with poorly controlled diabetes are at high risk of acquiring bacterial infections. However, conflicting results have been reported on neutrophil function in diabetes. We periodically evaluated neutrophil dysfunction in multiple low-dose streptozotocin (STZ)-induced diabetic mice, and then evaluated the effects of troglitazone and other thiazolidinediones (TZDs) on the decline of neutrophil function. Zymosan was injected intraperitoneally and neutrophil infiltration and phagocytosis were evaluated. While phagocytosis of zymosan by peritoneal neutrophils was consistently reduced in diabetic mice, neutrophil infiltration was decreased on day 30, but increased on day 40 after STZ injection. The in vitro chemotactic and phagocytic activities of blood neutrophils in mice that did not receive zymosan were consistently reduced in diabetic mice. Phorbol myristate acetate (PMA)-stimulated superoxide production by zymosan-induced peritoneal neutrophils and the levels of zymosan-induced tumour necrosis factor (TNF)-α and interleukin (IL)-1β in peritoneal exudate fluids were also reduced in the diabetic mice. Treatment of the diabetic mice with troglitazone beginning 2 weeks after STZ injection did not improve hyperglycaemia but did prevent the decline of zymosan-induced neutrophil infiltration on day 30, and additionally promoted the increased infiltration on day 40. Troglitazone also promoted the chemotactic activity of blood neutrophils isolated from normal mice in vitro. Rosiglitazone but not pioglitazone induced a similar effect. Neutrophil phagocytosis was not enhanced by troglitazone either in vivo or in vitro. Taken together, neutrophil function is impaired by STZ-induced diabetes, but inflammatory infiltration does not always vary with the chemotactic disability or cytokine levels. Furthermore, troglitazone and rosiglitazone were suggested to improve at least neutrophil chemotactic activity in these animals.
Osteoprotegerin (OPG) acts as an endogenous decoy receptor in tumour necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis of fibroblast-like synovial cells
Authors: MIYASHITA T , KAWAKAMI A , NAKASHIMA T , YAMASAKI S , TAMAI M , TANAKA F , KAMACHI M , IDA H , MIGITA K , ORIGUCHI T , NAKAO K , EGUCHI K
Published: July 2004

We examined the role of osteoprotegerin (OPG) on tumour necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis in rheumatoid fibroblast-like synovial cells (FLS). OPG protein concentrations in synovial fluid from patients with rheumatoid arthritis (RA) correlated with those of interleukin (IL)-1β or IL-6. A similar correlation was present between IL-1β and IL-6 concentrations. Rheumatoid FLS in vitro expressed both death domain-containing receptors [death receptor 4 (DR4) and DR5] and decoy receptors [decoy receptor 1 (DcR1) and DcR2]. DR4 expression on FLS was weak compared with the expression of DR5, DcR1 and DcR2. Recombinant TRAIL (rTRAIL) rapidly induced apoptosis of FLS. DR5 as well as DR4 were functional with regard to TRAIL-mediated apoptosis induction in FLS; however, DR5 appeared be more efficient than DR4. In addition to soluble DR5 (sDR5) and sDR4, OPG administration significantly inhibited TRAIL-induced apoptogenic activity. OPG was identified in the culture supernatants of FLS, and its concentration increased significantly by the addition of IL-1β in a time-dependent manner. Neither IL-6 nor tumour necrosis factor (TNF)-α increased the production of OPG from FLS. TRAIL-induced apoptogenic activity towards FLS was reduced when rTRAIL was added without exchanging the culture media, and this was particularly noticeable in the IL-1β-stimulated FLS culture; however, the sensitivity of FLS to TRAIL-induced apoptosis itself was not changed by IL-1β. Interestingly, neutralization of endogenous OPG by adding anti-OPG monoclonal antibody (MoAb) to FLS culture restored TRAIL-mediated apoptosis. Our data demonstrate that OPG is an endogenous decoy receptor for TRAIL-induced apoptosis of FLS. In addition, IL-1β seems to promote the growth of rheumatoid synovial tissues through stimulation of OPG production, which interferes with TRAIL death signals in a competitive manner.
Prospective immunological profiling in a case of immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX)
Authors: BAKKE A C , PURTZER M Z , WILDIN R S
Published: July 2004

IPEX syndrome is a genetic autoimmune disease characterized by immune-mediated polyendocrinopathy, enteropathy, and X-linked inheritance. We describe a case of IPEX in which lymphocyte phenotypes were assessed at birth, before initiation of Cyclosporin A therapy, and at frequent intervals to 18 months of age. We performed flow cytometry for lymphocyte subtypes and for activation markers (HLA-DR, CD25, and CD69 or CD71). The ratios of both T to B cells and CD4+ to CD8+ cells were elevated at birth, but CD4+ cells were not activated. HLA-DR+ and CD25+ activated T-cells increased in association with two episodes of clinical deterioration: colitis and the onset of type I diabetes mellitus. These results indicate that measures of activation, particularly HLA-DR+ and CD25+ frequency, correlate well with the development of early active disease and may presage clinical episodes. Continuous maintenance of immunosuppression, once started, appears critical for prevention of permanent tissue damage.
Multiple antigenic peptides facilitate generation of anti-prion antibodies
Authors: Bainbridge J , Jones N , Walker B
Published: July 2004

Recent reports have demonstrated the ability of anti-prion antibodies to inhibit PrPSc propagation. Due to the relatively poor immunogenic properties of both PrPC and PrPSc, the generation of anti-prion antibodies still causes a significant problem in the development of immunotherapeutic strategies. This study examines the potential of multiple antigenic peptides (MAPs) to raise an antibody response to prion derived sequences in mice. The MAP was constructed of a four spiked ring. Two spikes containing human or mouse derived prion amino acid sequences and two spikes containing the universally promiscuous tetanus toxoid sequence (aa 830–844) which was used to assist T-cell-dependent B-cell antibody production. Following vaccinations with the MAP or MAP plus adjuvant, sera were taken and antibody titres assessed. The MAP containing only the mouse sequence failed to elicit a significant antibody response. MAPs containing human prion sequences elicited antibody production to the corresponding prion sequence. Further analysis also demonstrated that these peptides were able to generate antibody responses that recognize conserved human and mouse sequences. These homologous sequences contain the heralded PrPSc specific sequence ‘Tyr-Tyr-Arg’ and therefore these MAPs may have some therapeutic potential.
Prevention of experimental autoimmune encephalomyelitis in DA rats by grafting primary skin fibroblasts engineered to express transforming growth factor-β1
Authors: Zargarova T , Kulakova O , Prassolov V , Zharmukhamedova T , Tsyganova V , Turobov V , Ivanov D , Parfenov M , Sudomoina M , Chernajovsky Y , Favorova O
Published: July 2004

To determine whether primary fibroblasts producing latent transforming growth factor β1 (TGF-β1) are capable of down-regulating experimental autoimmune encephalomyelitis (EAE), a retroviral vector TGF-β1-pBabe-neo (−5′UTR) was used for efficient gene transfer into primary skin fibroblasts of DA rats. After heat activation, conditioned medium from the transduced fibroblasts was found to inhibit significantly in vitro proliferation of lymphocytes from lymph nodes of DA rats with EAE. Intraperitoneal administration of TGF-β1-transduced fibroblasts into DA rats during the priming phase of EAE resulted in a significant reduction in mortality and in the mean clinical and EAE scores versus the control immunized animals treated with non-transduced fibroblasts.
Escherichia coliup-regulates proinflammatory cytokine expression in granulocyte/macrophage lineages of CD34+ stem cells via p50 homodimeric NF-κB
Authors: KIM J M , OH Y-K , KIM Y-J , YOUN J , AHN M-J
Published: July 2004

Umbilical cord blood has emerged as an alternative source of haematopoietic CD34+ cells for allogeneic stem cell transplantation. Although bacteraemia induced by Escherichia coli is considered one of the complications of transplantation, expression of proinflammatory cytokines is poorly understood. In this study, we report the altered expression of proinflammatory cytokines in CD34+ cells and their in vitro cultured cells following E. coli infection. CD34+ stem cells and their cultured cells up-regulated expression of proinflammatory cytokines such as interleukin (IL)-1α, IL-6, IL-8 and tumour necrosis factor (TNF)-α after infection with E. coli. Expression of the proinflammatory cytokines was generated mainly by the granulocyte-macrophage lineages. E. coli infection activated the signals of p50/p50 nuclear factor-kappaB (NF-κB) homodimers and IκB kinase. Furthermore, inhibition of NF-κB activation lowered the up-regulated expression of the proinflammatory cytokines. These results suggest that CD34+ cells and their cultured cells infected with E. coli induce the expression of proinflammatory cytokines via the NF-κB pathway.
β-Defensin-3 and -4 in intestinal epithelial cells display increased mRNA expression in ulcerative colitis
Authors: FAHLGREN A , HAMMARSTRÖM S , DANIELSSON Å , HAMMARSTRÖM M-L
Published: July 2004

mRNA expression of two recently described human β-defensins (hBD-3 and hBD-4) in epithelial cells of normal small and large intestine and the impact of chronic intestinal inflammation on their expression levels was investigated. Intestinal specimens from patients with ulcerative colitis (UC), Crohn's disease (CD) and controls with no history of inflammatory bowel disease were studied. hBD-3 and hBD-4 mRNAs were determined in freshly isolated epithelial cells by real-time quantitative reverse transcription–polymerase chain reaction (QRT-PCR) and by in situ hybridization. The effect of proinflammatory cytokines on hBD-3 and hBD-4 mRNA expression in colon carcinoma cells was also investigated. Purified epithelial cells of normal small and large intestine expressed both hBD-3 and hBD-4 mRNA, with higher expression levels of hBD-3 mRNA. In situ hybridization revealed higher levels of mRNA expression in the crypt- compared to the villus/luminal-compartment. Interferon (IFN)-γ, but not tumour necrosis factor (TNF)-α or IL-1β, augmented hBD-3 mRNA expression. None of these agents stimulated hBD-4 expression. Colonic epithelial cells from patients with UC displayed a significant increase in hBD-3 and hBD-4 mRNA compared to epithelial cells of controls. In contrast, small intestinal epithelial cells from CD patients did not show increased expression levels compared to the corresponding control cells. Moreover, Crohn's colitis did not show increased expression of hBD-4 mRNA, while the data are inconclusive for hBD-3 mRNA. We conclude that the chronic inflammatory reaction induced in the colon of UC patients enhances hBD-3 and hBD-4 mRNA expression in the epithelium, whereas in CD this is less evident.
Isolation and functional analysis of circulating dendritic cells from hepatitis C virus (HCV) RNA-positive and HCV RNA-negative patients with chronic hepatitis C: role of antiviral therapy
Authors: TSUBOUCHI E , AKBAR S M F , MURAKAMI H , HORIIKE N , ONJI M
Published: July 2004

Hepatitis C virus (HCV) RNA has been localized in antigen-presenting dendritic cells (DCs) from patients with chronic hepatitis C (CHC). DCs from patients with CHC also exhibit impaired functional capacities. However, HCV RNA in DCs and functional impairment of DCs in CHC might be independent or interrelated events. Moreover, the impact of antiviral therapy on the functions of DCs in CHC is not well documented. In order to address these issues, we took advantage of antiviral therapy in these patients. Ten patients with CHC, expressing HCV RNA in circulating DCs, became negative for HCV RNA in circulating DCs after therapy with interferon-α and ribavirin for 4 weeks. The functions of DCs from HCV RNA+ patients (isolated before antiviral therapy) and HCV RNA– patients (isolated 4 weeks after antiviral therapy) were compared in allogenic mixed leucocyte reactions. In comparison to circulating DCs from normal control subjects, DCs from HCV RNA+ patients had a significantly decreased capacity to stimulate allogenic T lymphocytes (P < 0·01) and produce interleukin-12 (P < 0·05). However, the allostimulatory capacity of circulating DCs from HCV RNA– patients was several-fold higher compared to that of HCV RNA+ DCs from the same patient. DC from HCV RNA– patients also produced significantly higher levels of interleukin-12 compared to HCV RNA+ DCs from the same patient (P < 0·01). Taken together, this study is the first to provide experimental evidence regarding the impact of HCV RNA and antiviral therapy on the function of DCs in patients with CHC.
Anti-inflammatory effect ofViscum albumagglutinin-I (VAA-I): induction of apoptosis in activated neutrophils and inhibition of lipopolysaccharide-induced neutrophilic inflammationin vivo
Authors: LAVASTRE V , CAVALLI H , RATTHE C , GIRARD D
Published: July 2004

Viscum album agglutinin-I (VAA-I) is a plant lectin which possesses antitumoral properties. This lectin is also known for its immunostimulatory effects when used at low concentrations (1–100 ng/ml). We have demonstrated recently that VAA-I is a potent inducer of human neutrophil apoptosis in vitro when used at higher concentrations. The role of VAA-I on activated neutrophils has not so far been investigated and its potential proinflammatory properties in vivo are poorly documented. Herein, we demonstrated that VAA-I (1000 ng/ml) induces apoptosis in lipopolysaccharide (LPS)-treated human neutrophils in vitro as well as in murine neutrophils isolated from lipopolysaccharide (LPS)-induced neutrophil influx. Using this model, we found that administration of VAA-I (100 or 1000 ng/ml) did not induce an inflammatory response. However, when used at 1 or 10 ng/ml, VAA-I was found to significantly induce a transitory inflammatory response, based on an increased leucocyte infiltration (>98% neutrophils). Also, we found that VAA-I inhibits LPS-induced neutrophil influx when administered simultaneously with LPS. In such conditions, some characteristic apoptotic neutrophils were observed in the pouch. Unlike LPS, which increased the production of some cytokines, VAA-I (1 or 10 ng/ml) did not increase the production of tumour necrosis factor (TNF)-α, interleukin (IL)-1Ra, IL-1α, IL-β, IL-8, IL-10 or IL-12 (p70) in human neutrophils. We conclude that VAA-I possesses the ability to induce apoptosis of preactivated neutrophils at a concentration that does not induce a proinflammatory response. Moreover, we conclude that VAA-I can inhibit a LPS-induced proinflammatory response in vivo. These data may provide new clinical perspectives in future mistletoe therapy and favour its potential utilization based on anti-inflammatory activity that at first appears contradictory with its use as immunostimulant.
DNA vaccination encoding glutamic acid decarboxylase can enhance insulitis and diabetes in correlation with a specific Th2/3 CD4 T cell response in non-obese diabetic mice
Authors: GAUVRIT A , DEBAILLEUL M , VU A-T , SAÏ P , BACH J-M
Published: July 2004

DNA vaccination encoding β cell autoantigens has been shown very recently to prevent type I diabetes in non-obese diabetic (NOD) mice. However, DNA vaccination encoding microbial or reporter antigens is known to induce specific long-lasting CD4 Th1 and strong cytolytic CD8 T cell responses. As this immune phenotype is associated strongly with β cell destruction leading to diabetes, we have chosen to study the effects of plasmids encoding glutamic acid decarboxylase (GAD), a crucial β cell autoantigen, in female NOD mice that developed a ‘moderate’ diabetes incidence. In the present study, 3-week-old  female  NOD  mice  were  vaccinated  twice  in  tibialis  muscles  with  plasmid-DNA  encoding 65-kDa GAD or βgalactosidase. In GAD-DNA immunized mice, diabetes cumulative incidence (P < 3·10−3) and insulitis (P < 7·10−3) increased significantly. Simultaneously, DNA immunization induced GAD-specific CD4 T cells secreting interleukin (IL)-4 (P < 0·05) and transforming growth factor (TGF)-β (P = 0·03). These cells were detected in spleen and in pancreatic lymph nodes. Furthermore, vaccination produced high amounts of Th2 cytokine-related IgG1 (P < 3·10−3) and TGF-β-related IgG2b to GAD (P = 0·015). Surprisingly, diabetes onset was correlated positively with Th2-related GAD-specific IgG1 (P < 10−4) and TGF-β-related IgG2b (P < 3·10−3). Moreover, pancreatic lesions resembled Th2-related allergic inflammation. These results indicate, for the first time, that GAD-DNA vaccination could increase insulitis and diabetes in NOD mice. In addition, our study suggests that Th2/3 cells may have potentiated β cell injury.
Role of peptide antigen for induction of inhibitory antibodies toStreptococcus mutansin human oral cavity
Authors: TSUHA Y , HANADA N , ASANO T , ABEI T , YAMAGUCHI S , SALAM M A , NAKAO R , TAKEUCHI H , KUROSAKI N , SENPUKU H
Published: July 2004

The alanine-rich repeating region (A-region) in the surface protein antigen (PAc) of Streptococcus mutans has received much attention as an antigenic component for vaccines against dental caries. The PAc (residue 361–386) peptide in the A-region possesses a multiple binding motif (L- -V-K- -A) to various HLA-DR molecules and a B-cell core epitope (- Y- - -L- -Y- - - -) that recognizes the inhibiting antibody to S. mutans. In the present study, we investigated the immunogenicity of the PAc (361–386) peptide in humans and regulators of induction of the anti-PAc (361–386) peptide IgA antibody (aPPA) in saliva. The PAc (361–386) peptide was confirmed as an ideal peptide antigen for induction of the inhibiting antibody to S. mutans in 151 healthy human subjects (36·6 ± 12·6 years old) by quantitative analyses of oral bacteria and ELISA, as the aPPA titre in human saliva decreased significantly in an age-dependent manner. Homozygous DRB1*0405 and 1502, and heterozygous DRB1*0405/1502 showed a negative association with production of aPPA and tended to reduce the number of total streptococci in saliva. In contrast, the DRB1*1501 allele was significantly correlated with a high level of induction of the antibodies, and also tended to reduce lactobacilli and mutans streptococci. Further, peptide immunogenicity was confirmed in NOD-SCID mice grafted with human peripheral blood mononuclear cells. Our results indicate that the interplay between regulators such as age, DRB1 genotype, cytokines, and peptide immunogenicity may provide a potential means for developing a vaccine useful for the prevention of dental caries as well as their diagnosis.
Autoimmune polyglandular syndrome Type 2: the tip of an iceberg?
Authors: Betterle C , Lazzarotto F , Presotto F
Published: July 2004

Autoimmune polyglandular syndromes (APS) are conditions characterized by the association of two or more organ-specific disorders. Type 2 APS is defined by the occurrence of Addison's disease with thyroid autoimmune disease and/or Type 1 diabetes mellitus. Clinically overt disorders are considered only the tip of the autoimmune iceberg, since latent forms are much more frequent. Historical, clinical, genetic, and immunological aspects of Type 2 APS are reviewed. Furthermore, data on 146 personal cases of Type 2 APS are also reported.
Alternatively spliced transcripts of Fas mRNAs in feline lymphoid cells
Authors: Mizuno T. , Baba K. , Goto Y. , Masuda K. , Ohno K. , Tsujimoto H.
Published: July 2004

Fas belongs to the tumour necrosis factor receptor family and transduces the death signal after binding to the Fas ligand. Five feline lymphoma cell lines were shown, by reverse transcription–polymerase chain reaction, to express six species of Fas transcripts. Based on sequence comparison of these Fas transcripts with the genomic Fas gene, five of the six transcripts were found to be generated through alternative splicing and to encode five different Fas proteins lacking the transmembrane domain. We also detected such alternatively spliced transcripts in primary tumour tissues from cats with naturally occurring lymphoma. These results suggest a possible association of the alternatively spliced Fas variants with the pathogenesis of feline lymphoma.
Association between platelet endothelial cellular adhesion molecule 1 (PECAM‐1/CD31) polymorphisms and acute myocardial infarction: a study in patients from Sicily
Authors: Listì F. , Candore G. , Lio D. , Cavallone L. , Colonna‐Romano G. , Caruso M. , Hoffmann E. , Caruso C.
Published: July 2004

Adhesion of circulating cells to the arterial surface is among the first detectable events in atherogenesis. Cellular adhesion molecules, expressed by the vascular endothelium and by circulating leucocytes, mediate cell recruitment and their transendothelial migration. Platelet endothelial cellular adhesion molecule 1 (PECAM‐1/CD31), involved in this migration, has been associated with the developmental course of atherosclerosis. A few studies have investigated an association between coronary heart disease and single nucleotide polymorphisms (SNPs) located in functionally important domains of the PECAM‐1/CD31 gene. In particular, Ser563Asn and Gly670Arg SNPs have been described as susceptibility factors involved in acute myocardial infarction (AMI) in the Japanese male population. To confirm these observations, we studied 96 male patients (mean age 40 years; age range 20–46) affected by AMI and 118 healthy male controls (mean age 38 years, age range: 20–55), and analysed for the following PECAM‐1/CD31 SNPs: Val125Leu, Asn563Ser and Gly670Arg. The frequency of the Gly670Arg polymorphism was significantly higher in patients with AMI (58.9% vs. 48.3%; P = 0.019), whereas the frequencies of the other two SNPs (Leu125Val and Ser563Asn) were not significantly different between patients and controls. By comparing the observed number of 670Arg/Arg genotypes in the patients with the expected number, calculated from the allele frequency in a healthy population, a significance of P = 0.02 (odds ratio, 2.04; 95% CI: 1.1–3.7) was obtained, supporting a recessive model of inheritance. Hence, the differences between patients and controls are significant, but relatively small. However, as AMI is a multifactorial disease, any single mutation will only provide a small or modest contribution to the risk, which also depends on environmental interaction. All in all, we believe that the results of the present study would add support to the role of pro/anti‐inflammatory genotypes in determining susceptibility or resistance to immune‐inflammatory diseases, including atherosclerosis.
Effects of single nucleotide polymorphisms in the RANTES promoter region in healthy and HIV‐infected indigenous Chinese*
Authors: Zhao X.‐Y. , Lee S. S. , Wong K. H. , Chan K. C. W. , Ma S. , Yam W.‐C. , Yuen K. Y. , Ng M. H. , Zheng B. J.
Published: July 2004

We determined the occurrence of the single nucleotide polymorphisms (SNPs) −403A/G and −28C/G in the promoter region of RANTES in 1082 Chinese blood donors from northern and southern China and 249 HIV patients from southern China. Compared to healthy adults, Chinese AIDS patients had a significantly higher frequency of the −403G allele and haplotype I, −403G/−28C (P < 0.05), and a lower frequency of the −403A/A genotype (P < 0.01). Symptomatic patients had a higher frequency of the −28G allele and a lower frequency of the −28C/C genotype (P ≤ 0.01). The plasma RANTES level was significantly lower in blood donors homozygous for haplotype I than in those who were homozygous for haplotypes II and III (P < 0.05). The frequency of the −403G allele was found to be higher in Chinese than in indigenous Africans, but lower than in Caucasians, Hispanics, and African Americans. The frequency of the −28G allele was comparable in Chinese and Japanese; this allele is rare in other ethnic groups. Results suggest that −403G may be associated with increased susceptibility to HIV infection, while −28G may be associated with advanced disease progression. The impact of SNPs on HIV infection appears to be unique in Chinese.
HLA‐DRB1 genotyping in patients with juvenile idiopathic arthritis in Taiwan
Authors: Huang J.‐L. , Yeh C.‐C. , Shaw C.‐K. , Yao T.‐C. , Chen L.‐C. , Lee T.‐D. , Kuo M.‐L.
Published: July 2004

The object of this study was to investigate whether there is an association between HLA‐DRB1 alleles and the development of juvenile idiopathic arthritis (JIA) in Taiwan. HLA‐DRB1 alleles were studied in 60 patients with JIA and 200 healthy controls using polymerase chain reaction (PCR)/sequence‐specific oligonucleotide probes (SSO). The frequency of HLA‐DRB1*0405 in patients with JIA was found to be significantly higher than that in healthy controls [odds ratio (OR) 2.64, 95% confidence interval (CI) 1.01–6.91]. The DRB1*0405 allele was significantly associated with the development of both polyarthritis (OR 4.30, 95% CI 1.34–13.80) and oligoarthritis (OR 3.27, 95% CI 1.01–10.58). The frequency of HLA‐DRB1*1502 was higher in Taiwanese JIA patients with systemic arthritis than in controls (OR 18.09, 95% CI 2.25–145.73). We conclude that, in Taiwan, HLA‐DRB1*0405 is associated with the development of polyarthritis and oligoarthritis in children, and HLA‐DRB1*1502 is associated with the development of systemic arthritis.
Detection of polymorphisms in the tumour necrosis factor alpha candidate gene in sheep
Authors: Alvarez‐Busto J. , Ruiz‐Nuñez A. , Mazón L. I. , Jugo B. M.
Published: July 2004

Primers based on GenBank sequences of the ovine tumour necrosis factor (TNF)‐α gene were designed to amplify a 273‐bp fragment comprising part of the fourth exon and the 3′ untranslated region (UTR) of the ovine TNF‐α gene. Five different single‐strand conformational polymorphism (SSCP) patterns were detected in a number of unrelated animals and three different alleles were identified and sequenced. These alleles differed in one deletion and one single nucleotide polymorphism (SNP) and were named TNF*01, TNF*02 and TNF*03. These alleles corresponded to three sequences previously characterized by other groups. In the population analysis, no significant differences were found in the frequencies of the Latxa and Rasa breeds. This is the first description of allelic variation in the ovine TNF‐α gene.
Invertebrate community structure in streams of the Manawatu–Wanganui region, New Zealand: the roles of catchment versus reach scale influences
Authors: Death Russell G. , Joy Michael K.
Published: July 2004

1. Invertebrate communities at 187 least impacted streams in the Manawatu–Wanganui region of New Zealand were sampled between February and May 2000 to investigate the relative influence of catchment and reach scale environmental influences on community structure.

2. Of the 138 biological (fish and periphyton), local habitat and catchment scale descriptors used to examine invertebrate community patterns, alkalinity and conductivity were the most consistently influential predictors.

3. Of the 52 geographical information system (GIS)‐derived catchment variables (catchment geology, catchment land use, rainfall and topography) only per cent catchment in pasture, indigenous forest, coastal sand, crushed argillite and wind blown sand were associated with any measures of the invertebrate communities.

4. Grouping of communities based on GIS data in general, did not generate distinct community types. Groupings based on river catchment, conductivity and alkalinity however, did produce distinct communities.

5. Streams with very low alkalinity were dominated by Ephemeroptera, Plecoptera and Trichoptera that were gradually replaced by Mollusca, Crustacea and Chironomidae as alkalinity increases.

6. Habitat characteristics measured at the scale of the reach were more closely linked with measures of invertebrate community structure than any GIS derived variables or river classifications.