bluuebunny/crossref_metadata_2025 huggingface dataset:
Distribution and Nucleotide Biases of the Somatic Hypermutations in the Functional Kappa Light Chain Gene of a Human Follicular Lymphoma Line
Authors: WU HONGYAN , KAARTINEN M.
Published: November 2003

The immunoglobulin kappa chain gene of human lymphoma cell line HF‐1.3.4 was partially sequenced from the 3′ end of the leader exon 2.0 kb downstream. The sequenced stretch of DNA included 1.5 kb of the non‐coding JK region 3′ to the JK2 element of the mature gene. Among the known VK germline genes the closest relative was KV328, which was 91% homologous to HF‐1.3.4. In the 1.5 kb JK region homology with JK allele of Whitehurst et al. (allele 2) was 89%, with the allele of Hieter et al. (allele 1) 87%. The vast majority of the differences located in the leader intron, the VJ exon or 0.6 kb 3′ to the exon, a localization characteristic of somatic hypermutations of immunoglobulin genes. Another indication that most of the differences observed were due to somatic hypermutations is that the 153 bp stretch of the kappa constant gene (CK) sequenced from the mRNA was 100% homologous with the published CK sequence. The most differences between the JK region sequence and that of Whitehurst et al. probably represent somatic mutations: 43% were transversions, 55% transitions and 2% deletions. In the non‐coding JK region transversions of C.G to G.C rather than to A.T were heavily over‐represented. This is possibly a feature of B‐cell hypermutations in humans and mice.
Activation of Cloned Human CD4+ Th1 and Th2 Cells by Blood Dendritic Cells
Authors: ROTH R. , SPIEGELBERG H. L.
Published: November 2003

Dendritic cells (DC) have been reported to be the most potent antigen‐presenting cells (APC) for the activation of naive T cells and to be 10–100‐fold more potent APC than monocytes (Mφ) in the mixed lymphocyte reaction. In this study the authors compared human blood DC with Mφ and B cells for their ability to activate cloned rye grass allergen Lol p I specific CD4+ Th1 and Th2 cells. In the presence of Lol  p I, all three types of APC activated Th1 and Th2 cells to a similar extent, as shown by T‐cell proliferation and interferon‐γ, interleukin‐2 (IL‐2) or IL‐4 secretion. However, at low APC : T cell ratios, Mφ were the most potent APC for both Th1 and Th2 cells followed in decreasing order by DC and B cells. This hierarchy was observed with APC preparations isolated by negative selection or highly purified by positive selection using fluorescent cell sorting for HLA‐DRhigh‐DC, CD14pos‐Mφ and CD19pos‐B cells. The data demonstrate that, in contrast to what has been reported for naive T cells, human blood DC activate cloned memory Th1 and Th2 cells to a similar extent as Mφ and B cells presumably because the requirements for activation of memory type T cells are less stringent than those for naive T cells.
The Effects of Mitogens, IL‐2 and Anti‐CD3 Antibody on the T‐Cell Receptor Vβ Repertoire
Authors: JASON J. , INGE K. L.
Published: November 2003

Phytohaemagglutinin (PHA), Concanavalin A (Con A), interleukin‐2 (IL‐2), and monoclonal antibodies to CD3 (CD3MoAbs) are used for the assessment of the T‐cell receptor (TCR) BV gene family expression in autoimmune disorders and multiple sclerosis, and to produce clones for assessment of cytokine profiles in progressive human immunodeficiency virus infection. The authors examined the effects of these stimulants on the TCR Vβ repertoire of resting and blastic CD4+ and CD8+ normal human peripheral blood lymphocytes, using three‐colour cytofluorometry and a panel of anti‐TCR Vβ monoclonal antibodies. IL‐2 was associated with an increased percentage of blastic CD4+ cells expressing Vβ5.1 (from median of 3.7% to 8.0%, P = 0.0002) and blastic CD8+ cells expressing Vβ5.3 (1.0 to 1.5%, P = 0.0039). CD3MoAb caused a slight increase in Vβ6.7 + blastic CD4+ cells (4.5 to 6.9%, P = 0.0078). PHA did not alter the Vβ repertoire of blastic cells. Con A caused skewing in CD8+ blastic cells, toward expression of Vβ5.2/5.3 (3.1 to 8.1%) and Vβ5.3 (0.8 to 4.8%) (P = 0.0020). Thus, IL‐2 stimulation causes slight alterations in the Vβ repertoire that should be taken into account in certain research settings. Con A produced skewing in CD8+ blastic cells suggesting that, in the presence of CD8, either Con A binds selectively to certain Vβ or the three‐dimensional complex created by Con A's binding to other T‐cell surface molecules induces preferential Vβ5 stimulation.
Bacteria‐Specific T‐Cell Clones are Selective in their Reactivity Towards Different Enterobacteria or H. pylori and Increased in Inflammatory Bowel Disease
Authors: DUCHMANN R. , MÄRKER‐HERMANN E. , MEYER ZUM BÜSCHENFELDE K.‐H.
Published: November 2003

In the present study the authors investigated the T‐cell response to different enterobacteria or Helicobacter pylori and tested the hypothesis that the frequency of bacteria‐specific T cells is increased in the intestine of patients with active inflammatory bowel disease (IBD), i.e. Crohn’s disease (CD) and ulcerative colitis (UC). The analysis of a large panel of T‐cell clones (Tc) (n = 888) from peripheral blood, non‐inflamed and inflamed intestine from IBD patients and control individuals shows that both peripheral blood and intestinal T‐cell clones were selectively stimulated by either Salmonella typhimurium Yersinia enterocolitica 03, Escherichia coli or Helcobacter pylori sonicates, that only < 3% of all bacteria‐reactive Tc were crossreactive and that proliferation to bacterial sonicates was inhibited by anti‐MHC class II antibody. In addition, bacteria‐specific Tc from IBD patients were more frequently isolated from inflamed intestine than from peripheral blood (P = 0.0039) or non‐inflamed intestine. These data, from a large number of T‐cell clones, are the first systematic analysis describing the response of individual T cells towards different bacterial species (ssp.). They show that T cells with specificity for distinct antigens or superantigens that are characteristic for a defined bacteria ssp. are present in normal, and increased in inflamed, IBD‐intestine. These bacteria‐specific Tc may play a role in IBD pathogenesis.
Signals Leading to the Activation of NF‐κB Transcription Factor are Stronger in Neonatal than Adult T Lymphocytes
Authors: KILPINEN S. , HENTTINEN T. , LAHDENPOHJA N. , HULKKONEN J. , HURME M.
Published: November 2003

The molecular background of the defects in the immune reactivity of human neonates has not been fully elucidated. As the NF‐κB transcription factor has a central role in the control of transcription of several genes involved in immune and inflammatory responses, the authors have analysed the activation of NF‐κB in human umbilical cord T lymphocytes. The activity was tested by quantitating the nuclear proteins binding to an oligonucleotide containing the consensus κB binding sequence (electrophoretic mobility shift assay). The data obtained demonstrate that phorbol dibutyrate/calcium ionophore A23187 (PDBu/iono) combination induced a clearly higher nuclear translocation of NF‐κB in neonatal than adult T cells. This higher NF‐κB activity was restricted to the CD4+ T‐cell subset. Analysis of the nuclear extracts with antibodies directed against the major components of NF‐κB the p50 and RelA (p65) proteins, indicated that the composition of NF‐κB was similar in neonatal and adult cells. These results suggest that neonatal T cells are exposed to oxidative stress‐inducing signals during delivery and/or are inherently more sensitive to NF‐κB activating signals than adult T cells.
Cloning of a Partial cDNA for Rat Interleukin‐12 (IL‐12) and Analysis of IL‐12 Expression In Vivo
Authors: MATHIESON P. W. , GILLESPIE K. M.
Published: November 2003

Experimental models of autoimmunity in the rat may feature selective activation of either the Th1 or Th2 subset of helper T cells. Interleukin‐12 (IL‐12) is a key cytokine in the development of Th1 responses. In order to study IL‐12 in the rat we used polymerase chain reaction (PCR) primers based on murine IL‐12 to amplify a partial cDNA from rat tissue. The product was cloned and sequenced: it shows 94% nucleotide identity with the murine gene and 94% identity of predicted amino acid sequence. Primers based on the rat IL‐12 sequence were used to analyse IL‐12 expression in vivo using semi‐quantitative PCR. We studied RNA from lymphoid tissues of two rat strains which differ in their response to mercuric chloride (HgCl2): Brown Norway (BN) rats develop autoimmunity with a predominant Th2 response; Lewis rats are resistant. Interleukin‐12 expression was higher in Lewis than BN, and higher in spleen than lymph node. After HgCl2, IL‐12 expression increased in BN towards the time when the autoimmune response autoregulates. Variation in baseline levels of IL‐12 expression may account for the Th2 predisposition of BN rats compared to Lewis rats; IL‐12 may play a role in the autoregulation of the Th2 response induced by HgCl2
Long‐Lasting Immune Response Induced by Recombinant Bacillus Calmette–Guérin (BCG) Secretion System
Authors: WADA N. , OHARA N. , KAMEOKA M. , NISHINO Y. , MATSUMOTO S. , NISHIYAMA T. , NAITO M. , YUKITAKE H. , OKADA Y. , IKUTA K. , YAMADA T.
Published: November 2003

The recombinant bacillus Calmette–Guérin (rBCG) secretion system utilizing an extracellular α antigen of Mycobacterium kansasii (α‐K) was characterized biochemically and immunologically. The human immunodeficiency virus type1 (HIV‐1) p17gag B cell epitope fused to α‐K was secreted in extremely large amounts. At least three mice out of seven inoculated with rBCG generated high titres of antibody to the epitope. The long‐lasting antibody production persisted more than 14 months.
Persistent CD3‐Crosslinking Down‐Regulates Interleukin‐2 Responsiveness in Interleukin‐2‐Competent Cloned T Cells: the Possible Involvement of Protein Kinase C
Authors: LANDEWÉ R. B. M. , DIJKMANS B. A. C. , VERDONK M.‐J. A. , BREEDVELD F. C. , DAHA M. R. , MILTENBURG A. M. M.
Published: November 2003

To investigate the regulation of interleukin‐2 (IL‐2) responsiveness of T cells, a human CD4+ T‐cell clone with constitutive expression of IL‐2 receptors was stimulated with recombinant IL‐2 (rIL‐2) in the presence or absence of immobilized anti‐CD3 monoclonal antibodies (αCD3imm MoAb). Incubation of T cells with αCD3imm MoAb decreased IL‐2‐induced proliferation which could not be ascribed to the modulation of IL‐2 receptor expression nor to cell death. Phorbol‐myristate‐acetate (PMA), an activator of protein kinase C (PKC), also induced down‐regulation of IL‐2 responsiveness. The αCD3sol MoAb, inducing Ca2+‐mobilization without activating PKC, did not inhibit IL‐2 responsiveness whereas cyclosporine A (CsA), a drug that inhibits the Ca2+‐dependent activation pathway, did not prevent the induction of IL‐2 hyporesponsiveness induced by αCD3imm MoAb. It is concluded that modulation of IL‐2 responsiveness of T cells via the T‐cell receptor/CD3 complex (TCR/CD3) may be mediated by a PKC‐activating signal.
Decreased Number of CD73 (ecto‐5′‐Nucleotidase) Molecules on Lymphocytes from Patients with Primary Immunoglobulin Deficiencies. Correlation between Number of CD73 Molecules and T‐Lymphocyte Function In Vitro
Authors: CHRISTENSEN L. D. , ANDERSEN V. , RYDER L.
Published: November 2003

CD73 is a bifunctional glycosyl phosphatidylinositol anchored leucocyte differentiation antigen which has specific ecto‐5′‐nucleotidase (ecto‐5′‐NT) activity and is an accessory T‐lymphocyte activation molecule. The aim of the present study was to investigate the CD73 expression on blood mononuclear cells (BMC) from a group of patients with primary immunoglobulin deficiency (IGD). This group of patients had both significantly decreased levels of ecto‐5′‐NT on BMC (P = 0.002) and decreased numbers of CD73 molecules per CD73+ lymphocyte (P = 0.01). Five of the 10 patients had a decreased percentage of CD73+ lymphocytes. Among B‐lymphocytes the patients had normal percentages of CD73+ cells but four of the 10 patients had numbers of CD73 molecules per CD73+ B‐lymphocyte below the normal range. Among CD4‐lymphocytes three out of 10 patients had percentages of CD73+ below the normal range and four out of 10 patients had decreased percentages of CD73+ CD8‐lymphocytes. Significant correlations were found between in vitro proliferative responses to mitogens and the number of CD73 molecules per CD73+ lymphocyte (rs = 0.60, P < 0.01) and per CD73+ CD8‐lymphocyte (rs = 0.64, P < 0.02). In addition, a positive correlation was found between ability to proliferate and level of ecto‐5′‐NT on BMC (rs = 0.53, P < 0.05). Furthermore the ability of BMC to synthesize ecto‐5′‐NT was studied. During 2 days' culture ecto‐5′‐NT activity increased markedly on BMC from both patients and healthy donors. The level of activity on BMC from all patients attained levels higher than on freshly isolated BMC from healthy donors. This shows that the decreased levels of ecto‐5′‐NT found on freshly isolated BMC from patients with IGD is due to defective regulation of the enzyme activity in vivo
Regulation of Expression of the Complement Factor H Gene in a Murine Liver Cell Line by Interferon‐γ
Authors: VIK D. P.
Published: November 2003

Factor H is a regulatory protein of the alternative pathway of complement activation that is synthesized mainly in the liver. The authors used the +/+ Li murine liver cell line as a model for examining its regulation. When +/+ Li cells were incubated with IFN‐γ, the levels of factor H mRNA increased in a dose‐dependent manner, achieving a maximal response at a concentration of 50–100 units/ml. The increase in factor H mRNA levels was paralleled by an increase in factor H secretion. The kinetics of induction of factor H mRNA were slow, with the response reaching near maximal levels at 24 h. The increase in factor H mRNA by IFN‐γ was dependent on protein synthesis, as cycloheximide abolished the response. The presence of IFN‐γ was required for the entire incubation period in order to produce a maximal response. The luciferase system was used in an attempt to identify an interferon‐responsive element. Luciferase constructs containing from 807 to 236 bp of upstream sequence responded to IFN‐γ with a twofold induction of luciferase activity, whereas a construct containing 83 bp of 5′ sequence did not. Thus, IFN‐γ stimulates factor H mRNA transcription through a protein intermediary that interacts with the promoter between positions −83 and −236.
Expression of MHC Class II Molecules on Human Monocytes is Regulated Independently from Each Other after Phagocytosis of Bacteria
Authors: WUORELA M. , JALKANEN S. , TOIVANEN P. , GRANFORS K.
Published: November 2003

Reactive arthritis is usually self‐limiting polyarthritis which develops in HLA‐B27 positive individuals after certain gastrointestinal or urogenital infections. The pathogenesis of reactive arthritis is unknown but T cells seem to have a crucial role. Most of the antigen‐specific T cells isolated from the synovial fluid have been MHC class II restricted. The role of antigen presentation in the pathogenesis of reactive arthritis has been studied relatively little. In this work the authors studied theeffect of arthritis‐triggering bacterium (Yersinia enterocolitica O:3) on the expression of MHC class II molecules on human monocytes and found that the expression of different MHC class II molecules was regulated independently from each other in half of the individuals after certain incubation periods. In these cases the expression of HLA‐DP was parallel to the expression of HLA‐DQ, while HLA‐DR expression went to the opposite direction or did not change at all. No difference between HLA‐B27negative and HLA‐B27 positive healthy individuals was seen. The authors conclude that independent regulation of the expression of different MHC class II antigens on antigen‐presenting cells is a more common phenomenon than usually thought.
Intranasal Inoculation of Bordetella Bronchiseptica in Mice Induces Long‐Lasting Antibody and T‐Cell Mediated Immune Responses
Authors: GUEIRARD P. , MINOPRIO P. , GUISO N.
Published: November 2003

Humoral and cellular immune responses were analysed in mice inoculated intranasally with Bordetella bronchiseptica. After infection, the number of bacteria that colonized the respiratory tract of the mice increased during the first day and decreased thereafter. Total IgG levels increased as early as 14 days after infection and decreased with time after infection, whereas total IgA and IgM levels were lower but remained stable. Specific antibodies to the bacteria were mainly IgG2a and IgA and persisted up to 10 months after infection. Some of these specific antibodies were directed against adenylate cyclase–haemolysin, the bacterial factor that had been shown to be necessary for initiation of infection. The proliferation of Bordetella bronchiseptica‐reactive spleen cells occurred during the acute phase of infection. T cells from infected mice produced increasing amounts of IFNγ and IL‐2 after infection. Although very low levels of IL‐10 were produced, no IL‐4 was detected after bacterial stimulation in vitro. These results suggest that Bordetella bronchiseptica infection induces primarily a Th1‐type T‐cell response. Importantly, the authors demonstrated that antibody and T‐cell responses directed against bacterial determinants of the virulent strain and to purified adenylate cyclase–haemolysin were long‐lasting. This observation could be due to the fact that Bordetella bronchiseptica may persist intracellularly in the host as it was demonstrated in vitro
Activity Inhibition of Cytolytic Lymphocytes by Omeprazole
Authors: SCARINGI L. , CORNACCHIONE P. , FETTUCCIARI K. , ROSATI E. , ROSSI R. , MARCONI P. , CAPODICASA E.
Published: November 2003

This study examined the in vitro effect of omeprazole (OM) on various types of murine cytocidal lymphocytes. The results show that OM caused a strong inhibition of basal natural killer (NK) activity in spleen cells (SC) from untreated CD2F1 mice; in peritoneal exudate cells and SC activated in vivo by injection of maleic anhydride divinyl ether 1,2‐copolymer (MVE‐2) or inactivated Candida albicans (CA); in lymphokine‐activated killer (LAK) activity generated in vitro from splenocytes cultured with rhIL‐2 and in allo‐specific cytotoxic lymphocyte‐mediated lysis generated in vitro. A significant inhibition of cytotoxic activity of all types of effector cells after 30 min incubation was already induced by OM at 1 × 10−3 m concentration, after 1 h incubation at 5 × 10−4 m and after 4 h incubation at 1 × 10−4 m OM. Complete inhibition of lytic activity was obtained after 4 h incubation of effector cells with 1 × 10−3 m OM. No inhibitory effect was observed at 5 × 10−5 m OM concentration. Indomethacin did not abrogate the OM inhibitory effect on NK/LAK activity, suggesting that prostaglandins are not involved in the process leading to suppression of cytocidal activity. When effector cells were incubated with OM in presence of rhIL‐2 (500 U/ml), the cytokine failed to antagonize the inhibitory effect of the drug. On the contrary, if OM pretreated cells were incubated with rhIL‐2 for a further 18 h after drug removal, this cytokine was able to restore NK activity, but only when NK inhibition was incomplete. These results demonstrate for the first time that in vitro OM causes a rapid, strong effect on various types of cytotoxic lymphocytes ranging from cytotoxicity inhibition to irreversible cell damage.
E‐Selectin: Sialyl Lewis, a Dependent Adhesion of Colon Cancer Cells, is Inhibited Differently by Antibodies Against E‐Selectin Ligands
Authors: SRINIVAS U. , PÅHLSSON P. , LUNDBLAD A.
Published: November 2003

Recent studies have demonstrated that selectins, a new family of cell‐adhesion molecules with similar domain structures, mediate the adhesion of peripheral blood cells to interleukin‐1 (IL‐1)‐activated endothelium. In the present study the authors evaluated the role of E‐selectin‐Sialyl Lewis x (SLex)/Sialyl Lewis a (SLea) interaction in mediating in vitro adhesion of two colon cancer cell lines, HT‐29 and COLO 201, to human umbilical cord endothelial cells (HUVEC). Colon cancer cell lines had a strong expression of blood group‐related carbohydrate epitopes as evaluated by fluorescence‐activated cell sorter (FACS) analysis. It was established that adhesion of HT‐29 and COLO 201 cells to IL‐1 stimulated HUVEC was calcium dependent and could be inhibited by a monoclonal antibody directed against E‐selectin. Prior incubation of cells with two different antibodies directed against SLex and antibodies directed against related Lewis epitopes, Lex and Lea, had no significant effect on adhesion. Three antibodies directed against SLea differed in their capacity to inhibit the adhesion of HT‐29 and COLO 201 cells to HUVEC. Only one antibody directed against the SLea structure was effective in inhibiting adhesion of both COLO 201 and HT‐29 cells. The difference could not be attributed to titre, the type or number of glycoproteins, or to a difference in the amount of SLea present on individual proteins, suggesting that presence and right presentation of SLea epitope might be important for adhesion of colon cancer cells. Finally, in the in vitro system used, adhesion of HT‐29 and COLO 201 cells to activated HUVEC is mediated predominantly by E‐selectin/SLea interaction. SLex and related epitopes, Lex and Lea, seem to have limited relevance for colon cancer cell recognition of E‐selectin.
Requirement of CD80 and CD86 Molecules for Antigen Presentation by Eosinophils
Authors: TAMURA N. , ISHII N. , NAKAZAWA M. , NAGOYA M. , YOSHINARI M. , AMANO T. , NAKAZIMA H. , MINAMI M.
Published: November 2003

The authors analysed the antigen‐presenting ability of eosinophils purified from peritoneal exudate cells of interleukin‐5 (IL‐5) transgenic mice. The granulocyte‐macrophage colony‐stimulating factor (GM‐CSF)‐treated eosinophils induced proliferative responses of primed lymph node T cells and thymus T cells to staphylococcal enterotoxin B (SEB), while untreated eosinophils induced little or no response. GM‐CSF‐treated eosinophils also induced proliferation of ovalbumin (OVA)‐primed lymph node T cells to OVA. Although untreated eosinophils expressed no MHC class II molecule on the surface the eosinophils could be induced to express major histocompatibility complex (MHC) class II molecules when treated with GM‐CSF. In the present study, anti‐I‐Ak monoclonal antibodies (MoAbs) strongly inhibited proliferation of thymus T cells and proliferation of OVA‐primed lymph node T cells in response to OVA, but weakly inhibited proliferation of primed T cells in response to SEB. Furthermore, CD80 (B7‐1) and CD86 (B7‐2) were expressed on the surfaces of untreated eosinophils. The expression of those two molecules on the eosinophils was increased by incubation with GM‐CSF. Moreover, anti‐CD80 or anti‐CD86 MoAbs blocked proliferative responses of primed lymph node T cells and thymus T cells to SEB, and also blocked responses of primed lymph node T cells to OVA. Thus, CD80 and CD86 play an important role in stimulation of T cells by eosinophils.
Regulation of α4 Integrin Avidity in Human B Cells: Requirement for Dephosphorylation Events for High Avidity VCAM‐1 Binding
Authors: Hedman H. , Lundgren E.
Published: November 2003

The authors compared the phosphorylation‐dependent signal transduction pathways involved in the regulation of adhesiveness of integrin LFA‐1 with that of α4 integrins binding to VCAM‐1. The authors developed an in vitro method to monitor changes in adhesiveness using a VCAM‐1 fusion protein coupled to magnetic beads. For LFA‐1, a similar method has previously been established using an ICAM‐1 fusion protein. Binding of cells was monitored and found to be strictly integrin α4 and VCAM‐1 dependent. The serine/threonine phosphatase inhibitors okadaic acid and calyculin A were equally potent in inhibiting binding to VCAM‐1 as to ICAM‐1, and inhibition of protein phosphatase‐1 (PP1) is proposed to be the important denominator. Similarly, the phorbol ester PDBu, potentially stimulating protein phosphatase‐1 and staurosporine, an inhibitor of serine/threonine kinases, enhanced adhesion to VCAM‐1 as has previously been shown for ICAM‐1. A major difference was that a significant portion of the binding to VCAM‐1 was not susceptible to inhibition by drugs while binding to ICAM‐1 could be completely inhibited. We propose that the adhesiveness of the α4 integrins for VCAM‐1 and of LFA‐1 for ICAM‐1 is regulated by similar or identical protein kinases and phosphatases.
The Self‐Reactive Antibody Repertoire of Normal Human Serum IgM is Acquired in Early Childhood and Remains Conserved Throughout Life
Authors: Mouthon L. , Lacroix‐Desmazes S. , Nobrega A. , Barreau C. , Coutinho A. , Kazatchkine M. D.
Published: November 2003

The authors have used a quantitative immunoblotting technique to analyse the antibody repertoire of IgM in cord blood and in the serum of young children, young adult males and aged males directed towards antigens in homologous tissues utilized as sources of self antigens. The reactivities of IgM with self antigens exhibited striking homogeneity and invariance among newborns. Self‐reactive IgM repertoires of children, young adults and aged males were markedly conserved among individuals and comprised most of the anti‐self reactivities that prevailed in neonates. Reactivities of IgM with bacterial antigens showed a high degree of homogeneity among newborns but were more diverse in children, young adults and elderly individuals. Diversity of IgM reactivities with self and non‐self antigens did not vary significantly with aging. Principal component analysis (PCA) and linear discriminant analysis (LDA) of the data discriminated between self‐reactive IgM repertoires of newborns and children, but failed to discriminate between repertoires of children, young adults and aged males. The data indicate that the self‐reactive antibody repertoire of IgM differentiates during the first years of life and remains relatively constant thereafter.
Participation of Vβ4+‐, Vβ7+‐, and Vβ11+‐T Lymphocytes in Haematogenously Acquired Staphylococcus aureus Nephritis
Authors: VERBA V. , TARKOWSKI A.
Published: November 2003

The most frequent pathogen in haematogenously acquired kidney infections in humans is Staphylococcus aureus. In order to characterize in situ the immunological patterns of septic nephritis we developed a murine model of this disease. A single intravenous injection of S. aureus producing toxic shock syndrome toxin‐1 resulted in high frequency of inflammatory kidney lesions. Histopathologically, both focal and diffuse inflammatory infiltrates were seen in kidney cortex and medulla. Immunohistochemical evaluation revealed high numbers of Mac‐1+ phagocytic cells as well as CD4+‐ and CD8+‐lymphocytes. The expansion of lymphocytes carrying T‐cell receptor Vβ chain 4, 7, and 11 families in the kidney was observed. Our results suggest that the haematogenously acquired kidney infection by superantigen‐producing staphylococci leads to migration, in situ activation and expansion of responding T‐lymphocyte subsets.
Chicken Joint Amyloid Protein is of the AA‐type. I. Characterization of the Amyloid Protein
Authors: LANDMAN W. J. M. , SLETTEN K. , KOCH C. A. M. , TOOTEN P. C. J. , GRUYS E.
Published: November 2003

Amyloid fibrils were extracted from deposits in joint tissue of heavy breed layers with spontaneous amyloid arthropathy and characterized as being of the AA‐type. Amino acid sequencing revealed a pattern quite similar to duck AA. Acute phase sera of chicken experimentally injected with Enterococcus faecalis showed a SAA‐protein like band cross reacting with anti‐chicken AA in immunoblot.
The Fibroblast Mitogenic Activity Released from Human Basophilic Cell Line KU812 is Separate from Tryptase and PDGF Expression
Authors: FORSBERG‐NILSSON K. , DUNDER U. , NILSSON K. , SCHWARTZ L. B. , NILSSON G.
Published: November 2003

The human leukaemia cell line KU812 has previously been used to study basophil differentiation. In this study the authors analysed the capacity of KU812 to produce the mast cell proteinase tryptase and to synthesize factor(s) mitogenic for fibroblasts. KU812 cells were treated with tetradecanoyl‐phorbol‐13‐acetate (TPA), conditioned medium from the human T‐cell line Mo (Mo‐CM), or cultured under serum free conditions. After 4 days the cells were analysed for cell growth, differentiation, content of tryptase, and secretion of fibroblast mitogenic activity. Mo‐CM and serum starvation increased the expression while TPA treatment down‐regulated the expression of FcεRI‐α chain. An increase in tryptase content in cell extracts was detected after 4 days of culture in serum‐free medium or in the presence of Mo‐CM. KU812 conditioned media was found to have a baseline expression of mitogenic activity on normal human foreskin fibroblasts that was increased after serum starvation or after treatment with TPA. Mast cell‐derived tryptase has previously been reported to be mitogenic for fibroblasts, but in this study the expression of tryptase did not correlate with the expression of fibroblast mitogenic activity in KU812 cells. Furthermore, affinity‐purified lung tryptase did not show any mitogenic activity. Platelet‐derived growth factor was also excluded. Although the factor(s) from KU812 cells stimulating fibroblast proliferation have not been identified, our results indicate that basophils may be potential producers of growth factors inducing fibroblast proliferation.
RT‐PCR Based Analysis of T‐Cell Receptor B Variable Region Gene Usage in Normal Human Breast Skin Resident T Lymphocytes (SRT)
Authors: AHANGARI G. , BERG A. , JEDDI‐TEHRANI M. , HALAPI E. , HAMMAR H. , WIGZELL H.
Published: November 2003

The skin interfaces directly with the external environment that contains innumerable infectious agents. Therefore, an appropriate and rapid immunologic response is required to preserve internal homeostasis. An essential feature of the ‘skin immuno system' (SIS) is the presence of substantial numbers of T cells in normal skin. The T‐cell receptor repertoire from normal human breast skin was analysed quantitatively and qualitatively by using PCR amplification of reverse transcribed RNA, T‐cell receptor BV3 and BV14 gene usage was increased in skin T lymphocytes in all individuals tested (n = 8) compared to peripheral blood CD4+ and CD8+ T lymphocytes from the same individuals. The T‐cell receptor junctional diversity analysed by high resolution gel electrophoresis showed skin T‐cell BV3 and BV14 gene usage to be predominantly polyclonal. Superantigen stimulation of T cells in human skin is considered a likely explanation of the present finding.
Increasing Cytotoxic Activity and Production of Reactive Oxygen and Nitrogen Intermediates by Peritoneal Macrophages During the Development of Multiple Organ Dysfunction Syndrome in Mice
Authors: JANSEN M. J. J. M. , HENDRIKS T. , HUYBEN C. M. L. C. , TAX W. J. M. , VAN DER MEER J. W. M. , GORIS R. J. A.
Published: November 2003

A major problem in the intensive care unit nowadays is the development of multiple organ dysfunction syndrome (MODS), a cumulative sequence of progressive deterioration of organ functions. While the pathogenic pathways of MODS remain to be elucidated, it is assumed that cells of the host defence system, especially the macrophages, are altered in their function. During the development of MODS it is assumed that macrophages are overactivated and that an exaggerated inflammatory response may contribute to its pathogenesis. In order to gain insight into the alterations of the functional status of the macrophage during the development of MODS, a series of macrophage functions was measured in the subsequent phases of zymosan induced generalized inflammation in mice. Male C57BL/6 mice received a single dose of zymosan intraperitoneally and groups of animals were killed after 2, 5, 8, and 12 days. Peritoneal macrophages were collected for in vitro assessment of the ADCC, the production of superoxide (O−2) and nitric oxide (NO), and complement mediated phagocytosis and intracellular killing of Staphylococcus aureus. A single intraperitoneal injection with zymosan resulted in a three‐phase illness. During the third phase the animals developed MODS‐like symptoms. Peritoneal cells from control animals produced very low to non‐detectable amounts of O−2 and NO, and the cytotoxic activity was also low. During the development of MODS, from day 7 onwards, the ability to produce O−2 and NO‐2 became strongly elevated, as did the cytotoxic activity. These findings are in parallel with the development of MODS whereas the phagocytic and killing capacity remained essentially unaltered. The changes found could be detrimental for the organism, thus possibly contributing to the onset and development of MODS.
The Transcription Factor Oct2A Enhances V(D)J Recombination in Fibroblasts
Authors: DÖBBELING U.
Published: November 2003

The influence of the transcription factor Oct2A on V(D)J recombination was investigated. It was found that  co‐transfection of Oct2A together with an artificial recombination substrate increased the rate of V(D)J recombination by a factor of 3.4 in the fibroblast cell line L4 which had been stably transfected with genomic  DNA containing the rag‐1 and rag‐2 genes and their flanking regulatory sequences. This effect, however, was not observed in the pre‐B‐cell line 38B9. The effect of Oct2A depends on the presence of its transactivation domains indicating that Oct2A increases the transcription of genes that increase the rate of V(D)J recombination. This effect can be reversed by additional co‐transfection of the glucocorticoid receptor. In contrast to L4 cells Oct2A and the glucocorticoid receptor had no effect in 38B9 cells. From these results one may conclude that the effect of Oct2A is masked in 38B9 cells by endogenous Oct2A and other  redundant pre‐B‐cell‐specific transcription factors and that it can only be seen in cells lacking these factors.
Synthesis of Cytokines During Tumour Development in Mice Immunized with the Mycobacterial Antigen Complex A60
Authors: MAES H. , COCITO C.
Published: November 2003

The authors have previously reported on the ability of A60, an immunodominant antigenic complex of Mycobacterium bovis BCG, to prevent cancer development in mice challenged with EMT 6 tumour cells. Such effect proved to rely on neoplastic cell lysis by cytolytic T lymphocytes and activated macrophages. The involvement of cytokines in triggering the immune response leading to tumour rejection is analysed in the present work. The synthesis of IL‐2, IFN‐α and TNF‐α was strongly increased in A60‐primed mice. Cancer development depressed the blood levels of these three cytokines. In vitro cultures of lymphocytes from lymph nodes and blood of A60‐primed mice produced higher levels of these cytokines in the presence of A60, as  compared to cultures lacking A60. Such effect was inhibited by co‐incubation of lymphocytes with EMT  6  tumour cells. In vitro cultures of macrophages yielded higher levels of TNF‐α in the presence of A60 and co‐incubation of these cells with EMT 6 tumour cells also inhibited TNF‐α production. The enhanced synthesis of IL‐2 and IFN‐α, which promote activation of cytolytic T lymphocytes and macrophages, accounts for the increased tumour cell lysis induced in vivo by A60. The A60‐promoted synthesis of TNF‐α is partly responsible for the latter effect. The inhibitory action of EMT‐6 tumour cells on cytokine synthesis is a powerful mechanism of tumour escape from the immune system’s control.
The MoAb‐VκIIIb Cross‐Reactive Idiotope on A27a (Humkv325) Encoded Kappa Chains Maps to Framework Region 3
Authors: POTTER K. N. , LI Y. C. , ABRAHAM G. N. , CAPRA J. D.
Published: November 2003

The monoclonal antibody MoAb‐VκIIIb binds a cross‐reactive idiotopic (CRI) determinant on light (L) chains encoded by the VκIIIb subgroup A27a (Humkv325) gene segment. The aim of this study was to localize the MoAb‐VκIIIb CRI. Mutational analyses involving region exchanges between a CRI‐positive VκIIIb chain and a CRI‐negative Vκ1 chain indicate that the MoAb‐VκIIIb CRI is located in framework region (FR) 3 of A27a (Humkv325) encoded L chains. CRI‐positive kappa chains unpaired with a heavy (H) chain are reactive with MoAb‐VκIIIb, indicating that the CRI is located on the kappa chain alone without involvement of H chain residues. Combinatorial antibodies composed of non‐parental L and H chain pairings are reactive with MoAb‐VκIIIb only when the L chain is A27a (Humkv325) encoded. The CRI, therefore, is not readily perturbed by H chain interactions. When the FR3 from a CRI‐positive kappa chain replaced the FR3 in a CRI‐negative lambda chain, the determinant was no longer detectable with MoAb‐VκIIIb. It is possible, therefore, to exchange regions between kappa chains from different families and retain the CRI structure, however the determinant is lost when placed in a more foreign background such as a lambda chain. These data more precisely define the interaction between MoAb‐VκIIIb and its CRI, and indicate that there are limits within which antibody FRs can be shuffled and still retain their native structural features.
The Effect of Strenuous Exercise, Calorie Deficiency and Sleep Deprivation on White Blood Cells, Plasma Immunoglobulins and Cytokines
Authors: BØYUM A. , WIIK P. , GUSTAVSSON E. , VEIBY O. P. , RESELAND J. , HAUGEN A.‐H. , OPSTAD P. K.
Published: November 2003

Moderate exercise appears to stimulate the immune system, but there is good evidence that intense exercise can cause immune deficiency. In the present study the authors examined the effect of continuous physical exercise (35% of VO2 max), calorie deficiency and sleep deprivation on the immune system of young men participating in a 5–7 days military training course. There was a two–three fold increase of neutrophils from day 1, the values remained high and decreased slightly at the end of the course. Monocyte counts also increased with a pattern similar to that of neutrophils. Eosinophils decreased to 30% of control and lymphocyte numbers decreased by 30–40%. All the major subgroups (CD4 T cells , CD8 T cells, B cells, NK cells) were reduced. Neutrophil function, as tested by measuring chemotaxis, was significantly stimulated during the first days of the course, in particular in the group with the lowest calorie intake. The mitogenic response of lymphocytes to PHA and Con A was variable, ranging from stimulation during one course to no effect in another course. Serum levels of immunoglobulins decreased significantly during the course. IgG was reduced by 6–7%, IgA by 10–20% and IgM by 20–35%. The authors found no changes of interleukin 1, 2 and 4 during the course, but a (12–20%) reduction (P<0.01) of interleukin 6 , and an increase (P<0.01) of granulocyte–macrophage colony stimulating factor. Altogether the results from the ranger course present a mixed‐up picture. The non‐specific phagocyte‐related immunity was enhanced. On the other hand, the data indicate that even a moderate physical activity, around the clock, caused significant suppression of a number of parameters reflecting the status of the specific, lymphocyte‐related immunity. It is noteworthy, however, that there was no significantly increased infection rate during the course or in the first 4–5 weeks thereafter.
Local Intravaginal Vaccination of the Female Genital Tract
Authors: WASSÉN L. , SCHÖN K. , HOLMGREN J. , JERTBORN M. , LYCKE N.
Published: November 2003

In a clinical trial the authors tested whether local intravaginal or oral vaccination would stimulate a mucosal immune response in the female genital tract. The whole cell/B subunit (CTB) oral cholera vaccine was used. Two groups of previously unimmunized volunteers were given three doses of vaccine at 2‐week intervals: a first group of seven women received oral immunizations and a second group of seven women were immunized locally in the genital tract by mixing the vaccine with a well defined gel, eldexomer, and applying it directly in the fornix of the vagina. The women were given the first vaccination on day 10 of the menstrual cycle. Sampling of peripheral blood and of cervical mucus (CM) using an Aspiglaire syringe was performed immediately prior to the first dose and at 8–10 days following the last immunization. The study showed that while only three of the seven orally immunized women responded with detectable IgA and IgG anti‐CTB antibodies in the genital tract, six out of the seven women in the locally vaccinated group responded with genital tract antibodies. The responses were also generally stronger and CM contained higher specific IgA and secretory component containing anti‐CTB titres in the locally vaccinated group. Of the orally vaccinated individuals all responded with increases in serum anti‐CTB IgG and 4/7 also exhibited specific IgA serum titres. By contrast, only 3/7 in the intravaginal group responded with increases in serum IgG and IgA anti‐CTB titers following immunization. The authors conclude that local intravaginal vaccination using a well‐defined gel appears to be the route of choice to stimulate immunity in the female genital tract.
Regulation of Newborn and Adult Monocytes by PGE2: Evidence of Comparable Differentiation Status
Authors: MURPHY F. J. , REEN D. J.
Published: November 2003

PGE2, which is produced in large amounts by monocytes, is considered to be an important autocrine factor in the regulation of the immune response. Altered sensitivity to PGE2 is known to reflect the differentiation status of monocytes/macrophages. Since newborn monocytes are thought to be less differentiated than adult peripheral blood monocytes, this study was designed to determine whether the diminished newborn monocyte function, relative to adult peripheral blood monocytes, might be the result of an altered autocrine responsiveness to PGE2. Phosphodiesterase activity as well as cAMP and TNFα levels in response to PGE2 were measured in newborn and adult monocytes, as an index of sensitivity to PGE2. Basal intracellular cAMP levels were reduced in cord monocytes compared with adult monocytes. Addition of increasing concentrations of PGE2 (10 m−10–10 m−6) caused similar fold increases in intracellular levels of cAMP for both populations. Lipopolysaccharide‐induced TNFα production was significantly reduced in a similar fashion for both cord and adult monocytes at high (≥ 10−7 m) PGE2 concentration. Phosphodiesterase activity was comparable in lysates from both populations. This study shows that PGE2 does not differentially influence newborn and adult monocyte responses and therefore that the diminished functional abilities of newborn monocytes is not due to altered autocrine responsiveness.
Primitive Cytokines and Cytokine Receptors in Invertebrates: the Sea Star Asterias rubens as a Model of Study
Authors: LEGAC E. , VAUGIER G.‐L. , BOUSQUET F. , BAJELAN M. , LECLERC M.
Published: November 2003

It has been previously demonstrated that the sea star axial organ is a primitive immune organ. Phagocytic, lymphoid‐like cells have been characterized with properties similar to those of vertebrates. There is also evidence for an invertebrate cytokine network because IL‐1 and TNF‐like activities are clearly demonstrable. In addition, the authors have previously described preliminary evidence for IL‐2‐like activity in the sea star. In the present report, the authors obtained evidence for the presence of IL‐1‐ and IL‐2‐like molecules on axial organ cells. More interestingly, the results suggested that sea star cells express structures similar to human receptors for IL‐1, IL‐2, IL‐6 and IFN‐γ.
Analysis of Murine IgG Isotype Galactosylation in Collagen‐Induced Arthritis
Authors: WILLIAMS P. J. , RADEMACHER T. W.
Published: November 2003

The galactosylation status of IgG from both control and arthritic DBA‐1 mice was determined by exoglycosidase sequencing and an anti‐GlcNAc monoclonal based ELISA. Two to three weeks after arthritis onset mice with collagen‐induced arthritis (CIA) showed a modest increase in IgG anti‐GlcNAc reactivity against controls (0.644 ± 0.080 and 0.530 ± 0.087, respectively, mean absorbance ± SEM n = 4) consistent with previous literature reports. However, the authors were unable to detect any significant changes in the galactosylation of purified IgG from arthritic and control mice using direct sequencing techniques (percentage G0 of 34.5% ± 1.9 and 37.7% ± 2.4, respectively, mean ± SEM n = 4). It has been demonstrated that a correlation exists between percentage G0 and anti‐GlcNAc reactivity for purified human IgG and sera samples, respectively; no such correlation was found for murine IgG and sera. The galactosylation of purified polyclonal murine IgG isotypes was analysed on a separate group of arthritic mice selected for their high anti‐GlcNAc reactivities. No significant differences were observed between control and arthritic mice. Immunoglobulin G isotype specific differences were found, with IgG1 exhibiting the highest percentage G0 (45–48%) followed by IgG2a (27–37%), IgG3 (20–32%) and IgG2b the lowest (13–17%). The percentage G0 and anti‐GlcNAc reactivity of purified IgG1 and IgG2b showed a narrow range of values when compared to those of IgG2a and IgG3 samples. Pooled sera from both arthritic and control mice was used to purify large quantities of IgG3, which on analysis revealed a fourfold increase in anti‐GlcNAc reactivity in the arthritic sample compared to control. Paradoxically these same IgG3 samples contained similar percentage G0 levels as determined by direct sequencing. The results suggest that IgG1 and IgG2b exhibit isotype selective oligosaccharide processing as little sample heterogeneity could be observed. These two purified IgG isotypes displayed a good correlation between percentage G0 and anti‐GlcNAc reactivity; this was in contrast to IgG2a and IgG3. Immunoglobulin G3 from arthritic mice may have G0 oligosaccharides selectively paired together with no net increase in percentage G0. This observation is discussed in detail as is the role of agalactosyl IgG in murine type II collagen‐induced arthritis.
Protein Kinase C Activation is Involved in Ultraviolet B Irradiation‐Induced Endothelial Cell ICAM‐1 Up‐Regulation and Lymphocyte–Endothelium Interaction In Vitro
Authors: FUNK J. O. , HOLLER E. , KOHLHUBER F. , UEFFING M. , BORNKAMM G. W. , KIND P. , EISSNER G.
Published: November 2003

Lymphocyte–endothelium interactions are pivotal steps in mediating inflammatory responses. The authors have analysed the influence of ultraviolet B (UVB) irradiation on intercellular adhesion molecule (ICAM)‐1 expression on cells of the human microvascular endothelial cell line (HMEC)‐1 and the intracellular signalling pathways involved. Flow cytometry revealed dose‐dependent ICAM‐1 up‐regulation with maximum induced expression 24 h after sublethal UVB irradiation of 10 mJ/cm2. While anti‐tumour necrosis factor (TNF)‐α antibodies or recombinant human interleukin (IL)‐10 did not influence this response, anti‐interferon (IFN)‐γ antibodies blocked the UVB‐induced ICAM‐1 up‐regulation. Significant induction of intracellular/membrane‐bound IFN‐γ was measured as early as 6 h post‐UVB. Since previous work has shown a differential role of protein kinase C (PKC) in cytokine induced ICAM‐1 expression, the effect of a selective  bisindolylmaleimide‐derived PKC‐inhibitor (GF109203X) was studied. Ultraviolet B‐induced ICAM‐1 up‐regulation was effectively blocked by the PKC‐inhibitor, whereas a PKA‐inhibitor was ineffective. Moreover, immunofluorescence analysis showed a radiation‐induced membrane translocation of PKC‐α, indicative of enzyme activation, in HMEC‐1 cells already 30 min post‐UVB. The functional relevance of the UVB‐induced ICAM‐1 expression and involvement of PKC in this process was demonstrated in an adhesion assay with peripheral blood mononuclear cells. In conclusion, UVB‐induced ICAM‐1 expression on human endothelial cells involves PKC‐dependent pathways and can be prevented by a PKC‐inhibitor. The use of PKC‐inhibitors as additive modulators in immune reactions may bear clinical potential. The mechanisms of IFN‐γ induction in endothelial cells by UVB deserve further investigation.