bluuebunny/crossref_metadata_2025 huggingface dataset:
CD3+ T Cells in Severe Combined Immunodeficiency (SCID) Mice. V. Allogeneic T Cells Engrafted into SCID Mice Do Not Induce Graft‐Versus‐Host Disease in spite of the Absence of Host Veto and Natural Suppressor Cell Activity
Authors: TSCHERNING T. , RUDOLPHI A. , REIMANN J. , CLAESSON M. H.
Published: June 2006

Intravenous injection of 106 to 107 non‐fractionated spleen ceils (SC) from C57BL,6 (B6. H‐2b) mice into completely allogeneic, immuno deficient H‐2d severe combined immuno deficiency (scid) mice leads to engraftment of allogencic donor T cells. Mice analysed in the tenth week post‐transfer had engrafted donor‐type CD4+ and CDS+ T cells in the spleens but showed no clinical evidence of graft‐versus‐host disease (GVHD). Transfer of allogeneic T cells engrafted in scid recipients did not induce GVHD upon i.v. injection into secondary scid recipients and lead in most recipients to engraftment of a pureCD4+ T‐cell population. Experiments were carried out to investigate the reason(s) for the lack of GVHD in recipient scid mice, i.e. the presence of allotolerance in the engrafted donor T cells. Scid spleen cells (SC) efficiently stimulated alloreactive responses of B6 T cells: scid SC stimulated H‐2d‐specific cytotoxic responses in a B6 antiscid mixed lymphocyte culture in vitro, and scid SC injected i.v. into B6 mice efficiently primed splenic cytotoxic lymphocyte precursors against H‐2d alloantigens. Moreover when assayed in vitro, no veto activity or natural suppressor activity was detectable in scid SC. These data demonstrate that tolerizing mechanisms currently believed to operate in vivo can not explain the fact that allogeneic T cells injected i.v. into immunodeficient scid mice become tolerized against host‐type alloantigens. Our results are discussed in the tight of clinical experience of allogeneic T‐cell transfer in scid infants.
Regulation and Functional Involvement of Distinct Determinants of Leucocyte Function‐Associated Antigen 1 (LFA‐1) in T‐Cell Activation In Vitro
Authors: DESROCHES C. VERMOT , RIGAL D. , ANDRÉONI C.
Published: June 2006

The expression of leucocyte function‐associated antigen 1 (LFA‐1) was studied by immunofluorescence method on human peripheral blood mononuclear cells (PBMC) stimulated by the phytohaemagglutinin lectin (PHA).

Monoclonal antibodies (MoAb) Mas 191c. IOT18 (directed against the β‐chain. 95 kDa. CDI8) and IOT16. SPVL7, MHM24 (identificating the α‐chain. 180 kDa, CD11a) were used, defining the CD 11a/ CD18' antibody panel.

By means of cross‐linking or competitive experiments, we showed that these antibodies recognized al least four distinct and spatially distant domains on the LFA‐t molecule.

Immunofluorescence analysis revealed that the up‐regulation of LFA‐1 expression was a late event, similar to the expression kinetics of the HLA DR and CD38 molecules, and followed the appearance of 0025 and CD71 molecules. Moreover, it was established that the LFA‐1 up‐regulation required mRNA and protein synthesis.

Functional activity comparison of the different anti LFA‐1 MoAb showed that the CDI la MoAb significantly inhibited the proliferation of lymphocytes stimulated by the phytohaemagglutinin lo various extents, as the LFA‐ 1α determinant identified, By contrast, the CD 18 MoAb did not influence strongly this cell process. We observed only n dim inhibitory effect with the CD18 MoAb recognizing an epitope common or very close to an LFA‐1 α determinant.

These results suggested that the LFA‐1 antigen was important, at a molecular level, in the regulation of T‐cell activation.
Characterization of Nickel‐Specific T Cell Clones
Authors: SILVENNOINEN‐KASSINEN S. , POIKONEN K. , IKÄHEIMO I.
Published: June 2006

Nickel is the major cause of metal‐induced allergic dermatitis. Twelve nickel‐specific T cell clones were used to investigate the cellular immune reactions occurring in nickel sensitivity. The selection between the alternative T cell receptors α and βγδ and two alternative Vβ genes (Vβ5 and Vβ8) were studied to see if nickel induces a selective pressure for clones bearing particular genes. Cell surface markers were studied by monoclonal antibodies and flow cytometry. Soluble mediators were measured by an ELISA method.

The clones used T cell receptor αβ genes but did not use Vβ5 or Vβ8. They were T helper clones with a primed memory marker (CD3+CD4+CD8−CD45RO+) and carried HLA‐DR. None of the clones secreted IL‐1α, all of them secreted IL‐2 receptor. Four clones secreted IL‐lβ, six IL‐4 and seven IL‐6, the peaks in IL‐2Rand IL‐6 secretion preceding 1L‐4 secretion. The clones helped immunoglobulin synthesis.

The clones from late effector phase of the nickel allergic reaction favours the use of T cell receptors αβ genes. Nickel‐specific clones were phenotypically indistinguishable but differed in soluble mediators produced.
Human Follicular Dendritic Cells: Isolation and Characteristics in Situ and in Suspension
Authors: PARMENTIER H.K. , VAN DER LINDEN J. A. , KRIJEN J. , VAN WICHEN D. F. , RADEMAKERS L. H. P. M. , BLOEM A. C. , SCHUURMAN H‐J.
Published: June 2006

Follicular dendritic cells (FDC) in human tonsils, either in situ in follicular germinal centres or isolated from tissue, were characterized by immunohistochemical. enzyme cytochemical and electron microscopical methods. Using polyclonal and monoclonal antibodies, expression of DRC‐1. Ki‐M4, HLA‐DR, CRI, Clq antigens, a macrophage marker, and surface IgG and IgM were found on isolated FDC and on FDC in situ. None of these reagents proved to be specific for FDC. e.g. the FDC‐directed antibodies DRC‐1 and Ki‐M4 labelled B lymphocytes in cytofluorography.

Enzyme cytochemical staining revealed activities of non‐specific esterase, acid α‐naphthylacetate esterase and ATPase in germinal centres and in freshly isolated FDC.

Immunohistochemistry demonstrated a weak expression of CD4 by a fraction of isolated FDC, which was confirmed by two‐colour immuno‐staining and immuno‐electron microscopy.
A Tumour Necrosis Factor Beta Gene Polymorphism in Relation to Monokine Secretion and Insulin‐Dependent Diabetes Mellitus
Authors: POCIOT F , Mølvig J. , WOGENSEN L. , WORSAAE H. , DALBØGE H. , BAEK L. , NERUP J.
Published: June 2006

HLA‐class III region genes may be associated with susceptibility to insulin‐dependent diabetes mellitus (IDDM). In this study an Ncol polymorphism of the tumour necrosis factor beta (TNF‐β) gene, which is positioned next lo the tumour necrosis factor alpha (TNF‐α) gene in the HLA class 111 region, was detected by restriction fragment length polymorphism (RFLP). This polymorphism has previously been reported lo be located in the TNF‐α gene. Caucasian HLA‐DR3.4 heterozygous IDDM patients (n=‐26) and DR‐matched healthy controls (n=19). as well as randomly selected IDDM patients (n = 27) and controls (n = 25) were studied. In addition four multiplex families (49 individuals) and eight HLA‐non‐identical sibpairs concordant for IDDM were analysed.

The TNF‐β gene RFLP analysis showed fragments of 5.5 kb and 10.5 kb, which behaved as alleles. In all groups there was a haplotype assignment of the TNF‐β 5.5‐kb allele to BS, DR3 haplotypes, and of the TNF‐β 10.5‐kb allele to B15.DR4‐positive haplotypes. The allelic and genotypic frequencies differed between DR3.4 IDDM patients and DR3,4 controls, and the DR3.4 control group differed significantly from the randomly selected control group (P < 0.0079), In HLA‐DR3,4‐atid DQw8‐positive persons, the DR3 haplotypes carried the 10.5‐kb allele ihrcL‐ times more frequently in IDDM patients than in controls, suggesting that the 10.5‐kb allele when present on DR3 haplotyes may contribute lo susceplibility to IDDM in DR3.4 heterozygous individuals, A contributory role of the Hl.5‐kballele in genetic IDDM susceptibility was supported by the sibpair analysis, in which all were TNF‐/1 identical. Five were 10,5 kb homozygous. and the remaining three pairs were 5.5.10,5 kb heterozygous.

Twenty‐five healthy and eight newly diagnosed IDDM patients were randomly selected to study the Escherichia coli lipopolysaccharides (LPS)‐purified protein derivate (tuberculin) (PPD)‐, and phytohaemagglutinin (PHA)‐stimulated monocyte (Mo) secretions of interleukin 1 bela (IL‐1/J)and TNb‐α in relation lo the Ncol TNF‐/f gene polymorphism. The LPS‐ and PHA stimulated Mo IL‐l/f and TNF‐a: secretions were significantly lower for the TNF‐β 5.5.10,5 kb heterozygous individuals than for TNF‐β 10.5 kb homozygous individuals. Furthermore, the Mo IL‐1β and TNF‐a secretions of IDDM patients were significantly higher than the Mo secretions of TNF‐β genolype‐matched healthy controls.

This study suggests an association between the 10.5 kb TNF‐β allele and IDDM, and demonstrates an association between monokine responses and TNF‐β genotypes. These observations may have implications for understanding the pathogenesis of HLA‐associated autoimmune diseases including IDDM.
Use of Size Fractionation of in Vitro‐Activated Human B Lymphocytes for Studies of Cell Cycle‐Dependent Growth Regulation
Authors: ERIKSTEIN B. K. , SCHWARZE P. , BLOMHOFF H. KIIL , FUNDERUD S. , STOKKE T. , SMELAND E. B.
Published: June 2006

Cell cycle progression of in vitro‐stimulated human B lymphocytes occurs a synchronously. In order lo allow detailed studies of growth Control G1. B cells Were stimulated with anti‐μ and low molecular weight B‐cell growth factor (LMW BCGF) for 50 h and subsequently separated into nine fractions of cells by means of centrifugal elutriation. As judged by volume profiles, activation antigen expression and DNA content, the cells in fractions t 4 were in early Io mid‐G1, while fractions 5 7 mainly contained cells in late G1. and fractions 8 9 contained cells mainly in Sand G2, Cells in fractions 5–7 had passed the commitment point, as demonstrated by a high spontaneous incorporation of [3H]thymidine when recultured in medium alone. Moreover S‐phase entry of these cells was largely unaffected by exogenous growth‐promoting or growth‐inhibitory signals. Cells in early (fractions 1 2) and intermediate fractions (fractions 3 4) showed a negligible spontaneous 3[H]thymidine incorporation, but a significant proportion of these cells progressed to S phase upon restimulation. Moreover, while IL‐4 or the anti‐CD40 MoAb G28‐ 5 potently stimulated cells in early and intermediate fractions, the responsiveness to LMW BCGF alone was obtained Just prior to the commitment point.
Lyme Arthritis: Oligoclonal Anti‐Borrelia burgdorferi IgG Antibodies Occur in Joint Fluid and Serum
Authors: CRUZ M. , HANSEN K. , ERNERUDH J. , STEERE A. C. , LINK H.
Published: June 2006

The antibody response to Borrelia (B.) burgdorferi, and to measles virus as control antigen, was analysed by agarose isoelectric focusing (AIF) and immunoblot of joint fluid and serum from 10 patients with Lyme arthritis and 10 controls with rheumatoid arthritis. Among the Lyme arthritis patients, six had oligoclonal anti‐B, burgdorferi IgG antibody bands in joint fluid and Corresponding serum, one patient had oligoclonal antibody bands in joint fluid only and also an elevated B. burgdorferi IgG in joint fluid to serum antibody ratio as evidence of intra‐joint production of specific antibodies, and the remaining three patients were negative for oligoclonal‐specific antibody bands. Absorption with B. burgdorferi imagen confirmed the specificity of the ofigoclonal antibody bands. They comigrated only partially on AIF with oligoclonal bands of total IgG and the specificity of most oligoclonal IgG in joint fluid and serum in Lyme arthritis remains undefined. Among the controls, no anti‐B, burgdorferi IgG antibodies were detected by AIF and immunoblot. Instead. 9 of the 10 rheumatoid arthritis patients had oligoclonal anti‐measles IgG antibody bands which were restricted to the joint fluid in three of them, indicating local production. We conclude that Lyme arthritis is often accompanied by an oligoclonal specific antibody response in joint fluid and serum simultaneously, and occasionally by intrasynovial synthesis of oligoclonal‐specific antibodies.
Conglutinin Binds The HIV‐1 Envelope Glycoprotein gpl60 and Inhibits its Interaction with Cell Membrane CD4
Authors: ANDERSEN O. , SØRENSEN A.‐M. , SVEHAG S.‐E. , FENOUILLET E.
Published: June 2006

The highly glycosylated envelope glycoprotein (gpl60) of human immunodeficiency virus (HIV) interacts with the CD4 molecule present on the membrane of CD4+ cells and is involved in the pathobiology of HIV infection. Lectins bind glycoproteins through non‐covalent interactions with specific hexose residues. The mammalian C‐type lectin bovine conglutinin was examined for its ability to interact with recombinant gpl60 (rgpl60) produced in vaccinia virus‐infected BHK21 cells. Specific binding of conglutinin to rgp160 was demonstrated by ELISA. The interaction of bovine conglutinin with rgp160 was calcium‐dependent, which is characteristic of the binding of a C‐type lectin to its ligand, and the binding was inhibited in a dose‐dependent manner with A‐acetyl‐D‐glucosamine. Deglycosylation of rgpl60 abrogated the conglutinin binding. In addition, conglutinin exerted a dose‐dependent inhibition of the binding of rgp160 to the CD4 receptor on CEM 13 cells, as demonstrated by FACS analyses. These results indicate that conglutinin may inhibit the infection with HIV‐1 through its interaction with the viral envelope glycoprotein.
The Variability of Immunoglobulins and Albumin in Salivary Secretions of Children
Authors: GLEESON M. , DOBSON A. J. , FIRMAN D. W. , CRIPPS A. W. , CLANCY R. L. , WLODARCZYK J. H. , HENSLEY M. J.
Published: June 2006

The variation of concentrations of immunoglobulins and albumin in consecutive daily collections of saliva was studied m 33 infants, aged 6 months to 5 years, for periods ranging from 16 to 26 days. The concentration and the within‐child variability of IgA and albumin and the detection of IgG and IgM in saliva increased with age. Between‐child variances were greater than the within‐child variances by a factor of 2.8 for log (IgA) and 1.3 for log (albumin). The geometric mean IgA levels were consistently higher and IgG was detected more frequency during upper respiratory tract infections compared with periods of non‐infection. There were no changes in albumin levels between infection and non‐infection periods, suggesting a local immune response rather than serum leakage. There were significant within‐child correlations (autocorrelations) between levels of IgA in saliva collected on consecutive days and samples collected up to 3 days apart. The autocorrelations between levels of albumin were significant for samples collected up to 2 days apart. The autocorrelation for IgA was significantly greater during infection periods compared with non‐infection periods FOR samples collected on consecutive days.
D‐D Recombination Diversifies the CDR 3 Region of Chicken Immunoglobulin Heavy Chains
Authors: MANSIKKA A. , TOIVANEN P.
Published: June 2006

The occurrence of D‐D recombination during the embryonic differentiation of chicken B cells was studied, Ig heavy (H) chains were amplified by polymerase chain reaction from day‐12 bursal cDNA, and 30 random V‐D‐J regions were analysed by DNA sequencing. No gene conversion events were observed in any of the V regions, indicating that diversification of the H chains by gene conversion is not yet activated at this stage of embryonic B‐cell development. In contrast, the V‐D‐J joint regions were extremely heterogeneous. Most of the sequenced V‐D‐J joints were formed by direct joining of the single‐germline Vμ1 gene, one of the multiple‐germline D elements, and the single J gene. However, three V‐D‐J regions were clearly longer in size, and their D‐region structure indicated recombination between two or three different germline D elements. Thus, the present data suggest that D‐D recombination may have a role in diversification of the Ig H‐chain repertoire.
Influence of HIV Antigens on Functional Activity of Neutrophilic Granulocytes in
Authors: GABRILOVICH D. I. , KOZHICH A. T. , SUVOROVA Z. K. , IVANOV V. S. , MOSHNIKOV S. A. , CHIKIN L. D. , KOLEZONKOVA O. V. , POKROVSKY V. V.
Published: June 2006

The effect of nine HIV antigens, including eight synthetic peptides, on the functional activity of granulocytes was studied using the reduction of nitroblue tetrazolium test (NBT test). Some peptides partly suppressed the functional activity of granulocytes. The most pronounced suppression was caused by ImVL (HIV‐1 lysate immobilized on plates for ELISA) and SP‐7 (a synthetic peptide from the gp41 protein of HIV‐1). The degrees to which the functional activity of granulocytes was suppressed by ImVL and SP‐7 was in inverse proportion to the specific antibody concentrations. No correlation was found between the reduction in the NBT test value and the amount of CD4+, CD8+ cells on CD4/8 ratio.
Interferon‐Gamma Enhances PAF‐Acether Production by Stimulated Human Polymorphonuclear Leucocytes
Authors: GEFFNER J. R. , SCHATTNER M. A. , LAZZARI M. A. , ISTURIZ M. A.
Published: June 2006

Human polymorphonuclear leucocytes (PMN) stimulated with either immune complexes (IC), phorbol myristate acetate (PMA) or N‐formyl‐methionyl‐leucyl‐phenylalanine (FMLP) generate platelet‐activating factor (PAF‐acether), The present study demonstrates that treatment of PMN with recombinant human interferon‐gamma (IFN‐y) significantly enhanced the production of PAF‐acether by stimulated cells, in a concentration‐dependent mode. On the contrary, α and β IFN were completely unable to increase PAF‐acether synthesis by stimulated PMN. The significance of these results is discussed.
Regulation of Interleukin‐6 Production in Human Fetal Kupffer Cells
Authors: KUTTEH W. H. , RAINEY W. E. , CARR B. R.
Published: June 2006

Inflammatory mediators such asinterleukin‐1 beta (IL‐1β), tumour necrosis factor‐alpha (TNF‐α), and interleukin‐6 (IL‐6) exhibit local autocrine and paracrine effects as well as distant systemic eflecis on target cells. Human Kupffer cells, the fixed tissue macrophages of the liver, may modulate immune and endocrine function in early fetal development. We purified and cultured human fetal Kupffer cells to investigate the production of the cytokine. IL‐6. Fetal Kupffer cells treated with bacterial lipopolysaccharide (LPS) produced IL‐6 in a dose‐dependent fashion with maximal secretion (1000 pg per 106cells) observed within 12h using 10/μg of LPS/ml. Cortisol and dexamethasone, but not oestrogen, progesterone, or testosterone, dramatically suppressed the LPS‐stimulated secretion of IL‐6 by fetal Kupffer cells. None of the steroids tested altered basal production or enhanced the LPS‐stimulated production of IL‐6 by fetal Kupffer cells. The inhibition of glucococorticoids could be reversed by the addition of RU 486, indicating that this effect was mediated by the glucocorticoid receptor. These results demonstrate that the production of IL‐6 by fetal hepatic macrophages can be activated by LPS and suppressed by glucocorticoids. These studies suggest that Kupffer cells express mature macrophage function in early gestation and would be capable of regulatory roles in the growth and development of the fetus.
‘E‐Boxes’ as Promoter Elements in B Cell Lines and Untransformed B Lymphocytes
Authors: HÖGBOM E. , MAGNUSSON A.C. , LEANDERSON T.
Published: June 2006

The penta‐deca (pd) promoter element from the SP6 K promoter and the μE2 box from the Ig heavy chain intron enhancer were analysed in an electrophoretic mobility shift assay (EMSA), utilising cell extracts from total mouse splenic B cells before and after stimulation with lipopolysaccharide. With both probes a changed EMSA pattern was observed after stimulation of the cells, where higher molecular weight DNA/protein complexes became dominant. When the pd and μE2 sequence elements were cloned in front of a TATA box and analysed for their transcriptional promoting activity in lipopolysaccharide stimulated B cells, they were inactive. On the contrary, the same constructs were transcriptionally active in some but not all B cell lines. Thus, E‐boxes display functional heterogeneity as transcriptional activators depending on host cell.
Natural Killer Function in the Rat with Interstitial Nephritis
Authors: BRICIO T. , MAMPASO F.
Published: June 2006

Brown Norway (BN) rats with tubulointerstitial nephritis (TIN) have an augmented NK activity. Immunological manifestations of the disease indicate the existence of a highly activated B‐cell system, as reflected m the high levels of circulating antibodies and in the predominant spontaneous igG‐producing B‐cells in cultures. In addition, NK cells appear to induce positive signals for antibody production either when co‐cultured with B‐cells alone or after treatment with poly I.e. Our results suggest that NK cells might play a role in the immunoregulation of the humoral response in this disease.
Idiotype‐Specific, Major Histocompatibility Complex Restricted T Cells are of both Th1 and Th2 Type
Authors: LAURITZSEN G. F. , BOGEN B.
Published: June 2006

The lymphokine secretion patterns of seven independent 91–101.γ2315/1‐Ed specific. CD4+ T‐cell clones have been investigated. Six of the clones are of the Thl type as they secrete IL2 and IFNγ, but not IL4. Some of these SIX Thi clones produce TNF α/β, and some produce minor amounts of IL5 and IL6. One clone is of the Th2 type as it produces 1L4, IL5. and large amounts of 1L6. but not IL2, IFNγ or TNF. The Thl, Th2 classification does not have any stringent relationship to immunization protocol, fine specificity and Vα/Vβ gene segment utilization. The immunoregulatory significance of our findings for Id/MHC‐dependent T B cell interaction is discussed.
Capacity of CD8+ T Cells to Reject Immunogenic Variants of a Spontaneous Murine Carcinoma: Lack of Non‐Specific (NK1.1+) Effector Mechanisms
Authors: ELLIOTT B. E. , BARRON A. , MAXWELL L. , CARLOW D. A. , MACNAUGHTON S. , PROSS H.
Published: June 2006

Class I MHC‐expressing (Ia−) immunogenic (imm+) variants, which elicit a strong syngeneic host immune rejection response, can be isolated following 5‐azacytidine treatment from the MHC‐negative non‐immunogenic (imm−) murine carcinoma cell line SPI (10.1 subclone). in the present study, we have shown that CD4‐depleted CD8+ T cells are both necessary and sufficient for the rejection process. Treatment of semi‐syngeneic B6×CBA Fl mice with anti‐NK1.1+ antibodies had no effect on the rejection of immunogenic variants, although the splenic NK (natural killer) activity of recipients was fully abrogated. Thus NKI.I+ effectors, which include most NK and LAK (lymphokine activated killer) cells, are most likely not involved in the rejection process. This finding was supported by a complete lack of NK susceptibility of SPI cells in vitro, although variable killing by LAK and poly‐I:C‐induced killer cells was observed. To assess the role of NK1.1− LAK and other non‐T killers (e.g. cytolytic macrophages) in vivo, we determined the specificity of the rejection process. We examined the ability of immune animals to reject a mixture of non‐immunogenic parent tumour cells (or cells of an unrelated syngeneic tumour) and of the variant tumour cells used for the initial immunization. Growth of the parent tumour cells was unaffected while the same animals rejected the immunogenic tumour cells. Our findings support a primary role of tumour‐specific CD8+ T cells in the rejection of imm+ variants with no detectable involvement of non‐specific effector cells in the tumour destruction process.
Effect of Salmonella Bacteria on the Interaction of Human NK Cells with Endothelial Cells
Authors: PINOLA M. , SAKSELA E.
Published: June 2006

Leukocyte endothelial cell adhesion and its regulation are essential and complex initial aspects of lymphocyte migration. Various factors (IL‐1, TNF‐α, IFN‐γ etc.) have been shown to increase the endothelial adhesiveness for human lymphocytes, including natural killer cells (NK cells). In this work we have demonstrated that pretreatment of either the target endothelial cell monolayers or the binding LGL‐cells with mRS95. Salmonella Minnesota bacteria results in a substantial increase in the adhesiveness of LGL‐cells to endothelial cells. The increase was more prominent when the endothelial cells were treated than when the adhering LGL‐cells were similarly pretreated. The adhering cell population was significantly enriched with CD56 (Leu 19) and CD16 positive cells, i.e. cells with NK cell phenotype, when the lymphocyte population was pretreated. However, the pretreatment of EC resulted in a non‐specific increase in FC adhesiveness since the relative proportion of CD56 + (Leul9), CD16 + and CD3+ cells among the adhering cells did not significantly differ from the starting population. The bidirectional enhancement of adhesiveness of human NK cells to endothelium by mR595 Salmonella bacteria may be significant in the host defense responses against microbial infections.
Anti‐Interleukin‐6 Antibodies in Normal Human Serum
Authors: HANSEN M. B. , SVENSON M. , DIAMANT M. , BENDTZEN K.
Published: June 2006

High‐avidity IgG antibodies to the cytokine inlerleukin‐6 (IL‐6) were found in sera of apparently healthy adult individuals. These antibodies specifically interfered with an FLISA (enzyme‐linked immunosorbent assay) for IL‐6 in which specific polyclonal rabbit antibodies to human recombinant IL‐6 (rlL‐6) were used. Furthermore. using precipitation of 125‐I‐rIL‐6 with rabbit antibodies to human immunoglobulins (Ig). the sera of 7 out of 6S Danish blood donors were found to contain specific antibodies in substantial amounts. Judged by ELISA interference, gel filtration of sera incubated with 125I‐rIL‐6 and second antibody precipitation of 125I‐rIL‐6. IgG seemed to be the dominant IL‐6 binding protein in these normal sera. Using specific antibodies to human in light chains, it was found that the anti‐lL‐6 antibodies were of polyclonal origin. Moreover, there are at least two epitopes on the IL‐6 molecule, because more than one IgG bound lo some IL‐6 molecules at the same time, The anti‐IL‐6 antibodies did not cross‐react with a number of other human recombinant‐derived and native cytokines. The antibodies recognized native as well as rlL‐6. but preferentially monomeric lL‐6.
Identification of Anti‐Plasmodium falciparum Antibodies in Human Breast Milk
Authors: LEKE R. G. F. , NDANSI R. , SOUTHERLAND N. J. , QUAKYI I. A. , TAYLOR D. W.
Published: July 2006

Malarial infections are rarely observed in neonates. It has been postulated that some immunity may be passively transferred during nursing, but anti‐malarial antibodies (Abs) have not been detected m human milk. In this study, milk samples, collected 2‐14 days after parturition from women at the Central Maternity Hospital, Yaounde, were evaluated for total IgG and IgA antibody levels by radial diffusion, protein composition by SDS PAGE, anti‐malarial antibodies using an isotype‐specific immunofluorescence assay, and the ability to immunoprecipitate Plasmodium falciparum antigens metabolically labelled with 35S‐methionine. Results showed that anti‐P. falciparum antibodies were present in breast milk, and that paired milk and serum samples from individual women contained Abs that recognized similar malarial antigens.
Experimental Infection with a Haemorrhage‐Causing Trypanosoma vivax in N'Dama and Boran Cattle
Authors: WILLIAMS D. J. L. , LOGAN‐HENFREY L. L. , AUTHIÉ E. , SEELY C. , MCODIMBA F.
Published: July 2006

N'Dama cattle control experimental infections with clones of Trypanosoma congolense of varying degrees of virulence, but nothing is known about their capacity to control infections caused by highly virulent. East African stocks of T. vivax. Thus four N'Damas and four trypanosusceptible Borans were infected with a tsetse‐transmitted stock of T. vivax. IL2337. In Ayrshire cattle this stock is known to cause severe haemorrhagic disease.

No differences were observed in the parasitaemia between the two groups. Both groups became anaemic. The mean packed cell volume fell to 16.8 ± 5.0% in the N'Dama cattle and to 24.2 ± 2.2% in the Borans on day 26 post infection. These differences were not significant. Antibody responses to invariant trypanosome antigens were analysed. No differences were observed between the groups in the pattern of recognition or the isotype elicited. Antibody bound to the surface of erythrocytes was occasionally detected. No anti‐platelet activity was observed.

The results show that N'Dama cattle, which are known to be resistant to disease caused by T. congolense and by T. vivax stocks from West Africa, were highly susceptible to an infection of T. vivax which causes acute haemorrhagic disease.
Cell‐Mediated and Monoclonal Antibody‐Dependent Killing of Onchocerca volvulus Microfilariae*
Authors: TITANJI V. P. K. , NDE P. N. , MBACHAM W. F.
Published: July 2006

To identify the target antigens implicated in the adherence and killing of microfilariae (mf) by leucocytes, we incubated nodular mf and leucocytes in the presence of anti‐cuticular monoclonal antibodies (MOVs)and fresh scrum. Leucocyte donors were patients with a mean age of 37 years (with 0‐1 mf snip), who had lived in the endemic village studied rural least 10 yean After 16‐20h of incubation, up to 74% of the mf could be seen with 10 or more cell adhering, to them. By 36‐40 h up to 54% of the mf had been killed by the leucocytes in the presence of cocktail of monoclonal antibodies termed MOV GI V. The degree of killing in control experiments with the monoclonal antibody MOV I remained lower (P 0.05), ranging from 0.0 to 4.5% of mf with initial viability of 90‐95%. Western blotting revealed MOV GIV prominent target antigens of 10.5, 18.0. 23.5 and 27;kDa in crude surface of female O. volvulus. The detected antigens may play a role m host protection.
Evidence of Immunosuppression by Bovine Respiratory Syncytial Virus
Authors: WOLDEHIWET Z. , SHARMA R.
Published: July 2006

Respiratory syncytial virus (RSV) is a major respiratory pathogen in human infants and calves. Calves and lambs infected with bovine RSV show mild clinical signs but they are more susceptible to secondary infection with Pasteurella haemolytica. Lambs infected with P. haemolytica 6 days after experimental infection with bovine RSV had significantly greater magnitudes of fever, higher disease and lesion scores and higher mortality rates than those infected with P. haemolytica or bovine RSV alone (P < 0.05).

Experimental infection with bovine RSV is characterized by alterations in lymphocyte subpopulation sand down‐regulation of some of their functions. For example, the number of T helper cells is significantly reduced during the first week of infection and peripheral blood mononuclear cells obtained from bovine RSV‐infected lambs were less responsive to the mitogen phytohaemagglutinin but more susceptible to P. haemolytica cytotoxin than those obtained from control lambs. Infection with bovine RSV does not significantly affect the humoral immune responses of lambs against P. haemolytica cytotoxin. Bovine RSV does not appear to affect the capacity of alveolar macrophages to present antigens in vitro.
Tuberculosis and Leprosy: Attempts to Identify T‐Cell Antigens of Potential Value for Vaccine Design
Authors: KAUFMANN S. H. E. , GULLE H. , DAUGELAT S. , SCHOEL B.
Published: July 2006

Tuberculosis and leprosy are chronic bacterial infectious diseases which represent major health problems worldwide. It is generally accepted that, on the one hand, effective vaccination strategies are required for satisfactory control of these diseases and, on the other hand, that currently available vaccination measures are insufficient for this purpose. Ideally, a subunit vaccine should be designed which is composed of one or a few protective antigens. In this brief treatise our approach towards the identification of antigens with potential value for vaccine design is described. It comprises high resolution fractionation by two‐dimensional gel electrophoresis, transfer of separated fractions by electroelution and testing of separated fractions with viable T cells and accessory cells. Using this approach we find: (1) multiple antigens are recognized by T cells from leprosy and tuberculosis patients as well as healthy contacts; (2) apparently, suppressive antigens exist in leprosy; (3) an antigen cluster which is apparently indicative for immunity against M. tuberculosis is present among secreted proteins. We hope that further improvement of this methodology will help in the rational design of subunit vaccines against tuberculosis and leprosy.
Serodiagnosis of Bacterial Diseases: Problems and Developments
Authors: TERPSTRA W. J.
Published: July 2006

Serodiagnosis is useful when pathogens cannot be detected quickly or easily by conventional methods or have been eliminated by the patient. Taking the example of leptospirosis, the microscopic agglutination test to detect specific antibodies after leptospires have disappeared from the patient's bloodstream is of unsurpassed reliability. Unfortunately the test is used mainly in specialized laboratories that can culture leptospires to serve as live antigens. Freeze‐dried leptospires may allow the application of the test on a wide scale. The ELISA is useful for the detection of IgM antibodies in humans as a sign of current or recent infection. ELISA results must be confirmed by the microscopic agglutination test. A simplified easy‐to‐read ELISA using broadly reactive antigen for the detection of all types of leptospires might be useful for a quick detection of acute human leptospirosis. Using an appropriate antigenic preparation the ELISA is useful for the detection of antibodies in animals, e.g. cattle. As for pathogen detection, DNA‐based methods such as PCR are promising for an early, quick and specific diagnosis. Antigenic analysis by monoclonal antibodies and genetic analysis by restriction fragment length polymorphism are for the time being complementary methods for the characterization of leptospiral isolates.
Humoral Factors of Natural Resistance of Bos indicus Cattle Selected for Antibody Titre to Brucella abortus
Authors: GWAKISA P. S. , MINGA U. M.
Published: July 2006

The concentration of lysozyme, total immunoglobulin and bactericidal activity were measured in sera of Bos indicus cattle, retrospectively screened for specific antibodies to Brucella abortus and classified as being positive reactors or negative reactors. In addition, the effect of complement in the sera was studied to demonstrate complement dependence of antibody‐mediated bacterial killing. It was observed that, under the test conditions, serum bactericidal activity and concentration of total immunoglobulin were associated with high specific antibody levels (P < 0.001). Furthermore, there was a slight decrease in the lytic activity of lysozyme in the sera of animals with high antibody titres.
Anaplasmosis in Uganda. II. Prevalence of Bovine Anaplasmosis
Authors: SSENYONGA G. S. Z. , KAKOMA I. , MONTENEGRO‐JAMES S. , NYEKO P. J. , NANTEZA A. , BUGA R.
Published: July 2006

The prevalence of bovine anaplasmosis was studied in 320 Zebu cattle randomly selected from three regions of Uganda (central, south‐western and north‐western) using DOT‐ELISA, Western immunoblotting. Rapid Card Agglutination Test (RCAT), Capillary Tube Agglutination Test (CAT), Complement Fixation Test (CFT), and parasitological techniques. Dried blood on Whatman filter paper no. I was eluated in PBS 0.05% Tween 20 prior to testing at an initial dilution of 1:25.

The incidence of parasitaemia ranged from 25% in the central region to 35% in the northwestern region and the serological prevalence was lower in the central region and highest in the north‐west. Prevalence rates assayed by DOT‐ELISA and Western immunoblotting were 1.5‐fold greater than those tested with RCAT and 3‐fold greater than in CAT. The overall prevalence rates by DOT‐ELISA and Western immunoblotting compared favourably with CFT data.

The present data utilizing dried blood on filter papers indicate that there is a high prevalence of anaplasmosis in those regions of Uganda surveyed and it confirms our observations and those of others that collecting blood on filter papers is a suitable technique for large‐scale screening and for seroepidemiological studies.
Anti‐hCG Vaccines are in Clinical Trials
Authors: TALWAR G. P. , SINGH OM , PAL R. , CHATTERJEE N.
Published: July 2006

Two vaccines inducing antibodies against human chorionic gonadotropin (hCG) have completed Phase I clinical trials, indicating the reversibility and safety of these vaccines. One is currently in Phase II efficacy trials in women in three major centres in India. The available data suggest that the vaccine prevents pregnancy above antibody titres of 50 ng/ml hCG bioneutralization capacity.
Determination by Widal Agglutination of the Baseline Titre for the Diagnosis of Typhoid Fever in Two Nigerian States
Authors: MOHAMMED I. , CHIKWEM J. O. , GASHAU W.
Published: July 2006

This study was carried out in Borno and Plateau States of Nigeria to determine the baseline titre for the diagnosis of typhoid fever using a single Widal test. Of 172 patients with symptoms and signs of typhoid fever, 92.4% and 90.7% had reciprocal O and H antibody titres respectively of 160 or above. On the other hand, 95.3% and 66.3% of the 937 healthy control subjects had reciprocal O and H antibody titres respectively of 80 or less. The results of this study suggest that in Borno and Plateau States of Nigeria a reciprocal O antibody titre of 160 and above in persons with illness whose symptoms and signs are compatible with typhoid fever could be considered diagnostic using the single Widal test.
Reduced Natural Killer Cell Activity in Multi‐Drug Resistant Pulmonary Tuberculosis
Authors: RATCLIFFE L. T. , MACKENZIE C. R. , LUKEY P. T. , RESS S. R.
Published: July 2006

Cellular immune status in five patients with multi‐drug resistant pulmonary tuberculosis was investigated and compared with five matched controls with non‐resistant tuberculosis. A significant reduction in fresh natural killer (NK)‐cell activity was found in the resistant group (P < 0.005). There were no significant differences between the two groups in lymphocyte phenotype, proliferation or PPD‐specific cytotoxicity. Reduced NK‐cell function may play a role in the pathogenesis of multi‐drug resistant pulmonary tuberculosis.
Serum Interleukin (IL‐1β) in Patients with Diffuse Scleritis
Authors: PALEXAS G. N. , PUREN A. , SAVAGE N. , WELSH N. H.
Published: July 2006

Scleritis is a painful, destructive inflammatory eye disease. We measured serum IL‐1β) in ten patients with diffuse scleritis and in control subjects matched for age and sex. Patients with diffuse scleritis were found to have significantly elevated serum concentrations of IL‐1/β in comparison to normal subjects (P<0.02). The results suggest that IL‐l/β is associated with diffuse scleritis and may influence scleral breakdown in diffuse scleritis.
Cells from Healthy Non‐Exposed Individuals Produce Cytokines to Selected Fractions of Leishmania Promastigotes
Authors: MAASHO K. , AKUFFO H. O.
Published: July 2006

We are interested in cellular responses to antigens of parasites to which the cell donor has not been previously exposed and how such responses may influence parasite establishment. In order to characterize such responses we have used cells from unexposed healthy donors and analysed the lymphoproliferative response to various Leishmania aethiopica antigen preparations and the cytokines produced in the process. Peripheral blood lymphocytes were stimulated with SDSPAGE separated L. aethiopica antigen coupled to nitrocellulose particles. Fifteen of the 16 unexposed individuals tested had proliferative responses to either the whole or and the antigen‐bearing nitrocellulose fractions (NC fractions). Although the degree of response to the fractionated antigen varied in individuals, major stimulatory fractions were found in the high molecular weight region of 110‐80 kDa (fractions 3‐6) and low molecular weight region of 46‐18 kDa (fractions 12‐16). Substantial amounts of interferon gamma (IFN‐γ) and interleukin 2 (IL‐2) were present in the supernatants of cells stimulated with the whole unfractionated antigen. The potential relevance of such responses in resistance to Leishmania infection is discussed.
Identification of Genetic Susceptibility loci for Insulin‐Dependent Diabetes in sudan
Authors: MAGZOUB M. M. A. , STEPHENS H. A. F. , GALE E. A. M. , BOTTAZZO G. FRANCO
Published: July 2006

In this study we report, for the first time, the molecular analysis of HLA‐DR and DQ gene frequencies in a large cohort of well‐characterized type I (insulin‐dependent) diabetes mellitus (IDDM) patients (n = 12), and ethnically matched controls (n = 59) collected in sub‐Saharan Africa. High molecular mass DNA was prepared and analysed in Southern blots and by oligonucleotide typing. We have shown a strong positive association between IDDM and the Asp 57‐ DQBl allele *0201 (DQw2). A rare DR4, DQw2 haplotype was also identified at high frequency in the IDDM cohort. We can now confirm that the association between Asp 57‐ DQBI alleles and IDDM, previously reported in ethnically diverse cohorts collected in Western Europe, North America, and South Asia, is also present in an IDDM cohort collected in Africa.
Post‐Mortem HLA Tissue Typing of Retinal Pigment Epithelial Cells
Authors: ZAVAZAVA N. , WESTPHAL E. , DUNCKER G. , NÖLLE B. , MÜLLER‐RUCHHOLTZ W.
Published: July 2006

Retinal pigment epithelial cells (RPE) were derived from bulbi of cornea donors and maintained in culture. The timespan between donor's death and cell cultivation ranged from 2 to 122 h. The mean numbers of hours was 25.6 (n = 130). After IFN‐y stimulation, cells were serologically typed for class I and class II antigens. Unclear class I allospecificities were verified by one‐dimensional isoelectric focusing. Data from the serological typing of lymphocytes and those of the serological and biochemical typing of RPE from the same donors were compared in 22 cases. There was a discrepancy of less than 5%, whereby either the typing on lymphocytes could not identify some specificities declared as blanks or the RPE typing had failed to clearly define a specificity. Our data show that the strategy adopted here is very successful for tissue typing post mortem, thus increasing the number of available HLA‐matched corneas and consequently reducing the number of corneal graft rejections.
Production, Immunoanalysis and Selection of Monoclonal Antibodies for Studies of Polymorphism of Bovine MHC Class I Antigens
Authors: GWAKISA P. S.
Published: July 2006

Two monoclonal antibodies (MoAbs) were raised against bovine lymphocyte antigens of the major histocompatibility complex (MHC) for studies of the polymorphism of MHC class I antigens of African cattle. Immunoanalysis of lymphoid tissues and molecular weight data were used to characterize the epitopes seen by the MoAbs. Competitive binding assays indicated specificity of the MoAbs for two distinct epitopes. Application of the MoAbs for MHC typing of a cattle population showed that the epitopes were co‐expressed on the target MHC antigen but occurred independently on non‐target MHC antigens. It is concluded that, to obtain MoAbs for studies of MHC polymorphism, it is important to conduct large‐scale analysis using immunoassays and population studies.
Reactivity of some Monoclonal Antibodies Specific for Human Lymphocytes with Vervet Monkey Peripheral Blood Mononuclear Cells
Authors: OLOBO J. O.
Published: July 2006

Reactivities of some monoclonal antibodies to human lymphocyte surface antigens were tested on vervet monkey peripheral blood mononuclear cells (PBMC), using flow cytometry and immunoperoxidase techniques. A number of antibodies were identified which reacted with similar populations of lymphocytes from vervet monkeys. These included antibodies that define T cells, suppressor/cytotoxic cell subset, pan B cells, monocytes and MHC class I and II. A number of anti‐CD4 markers examined were unsuitable for use in the vervet system.
T‐Independent Polyclonal Activation of B Cells In Vitro by Immunoglobulin Binding Substance (IBS) from the Granary Weevil
Authors: TAMURA H. , MITSUHASHI M. , MORIKAWA A. , KUROUME T. , YAMASHITA T. , YOKOTA Y. , OHSAWA S.
Published: June 2006

Experiments are described for T‐independent polyclonal activation of B cells in vitro by the immunoglobulin binding substance (IBS) from the granary weevil. The affinity‐chromatographically purified IBS was used. IBS is a heat‐, alkali‐ and acid‐stable glucopeptide which is characterized by non‐specific immunoglobulin binding lo the Fab fragment. The purified IBS consists of three polymer homologues whose molecular weights are 12–14,000, 25–30,000 and more than 150,000 Da. IBS did not stimulate DNA synthesis by murine T cells, macrophages or plasma cells whereas it did stimulate that by mature B cells without any help from T cells or macrophages, IBS also stimulated both in vitro IgG production by spleen cells and in vitro sensitization of spleen cells by sheep red cells (SRBC). IBS was found to stimulate DNA synthesis by B cells mediated by binding to surface immunoglobulins of B cells. IBS is thought to be a useful amplifier for inducing human hybridomas and a valuable tool for examining mature B cells, both diagnostically and experimentally.
Regulation of the Immune Response to Hepatitis B Virus and Human Serum Albumin. III. Induction of Anti‐Albumin Antibody Secretion In Vitro by C‐Gene‐Derived Proteins in Peripheral B Cells from Chronic Carriers of HBsAg
Authors: HELLSTRÖM U. B. , SYLVAN S. P. E.
Published: June 2006

The circulatory pool of B cells from the majority (11/13) of chronic hepatitis B surface antigen (HBsAg) carriers contained sensitized B cells with the capacity to secrete IgG antibodies with specificity for human serum albumin (HSA), when stimulated withE, coli‐derived core protein at low concentrations in vitro. The IgG anti‐USA secretion was dependent upon and regulated by T cells, and optimal secretion was obtained at T/B‐cell ratios of 1.0–4.0, varying for different individuals. The level of anti‐HSA secretion was higher for patients with on‐going viral replication as assessed by hepatitis B virus (HBV)‐DNA in serum. Culture supernatants containing anti‐HSA antibodies also contained anti‐HBc antibodies, as detected by enzyme‐linked immunosorbent assay (ELISA) where the solid phase was charged withE. coli‐derived core protein, or the synthetic peptides corresponding to the 75–84 and 132–147 sequences in the C region of HBV, In contrast, IgG anti‐HBc (E. coli‐derived), but no anti‐MSA or anti‐HBc 75–84, 132–147 antibodies, were detected at similar T/B‐cell ratios in cell cultures from 56 individuals with naturally acquired immunity to hepatitis B. These data indicate that peripheral B cells from the majority of HB‐immune donors are sensitized lo unique (e.g. non‐albumin associated) structures in the nucleocapsid of HBV, while B cells in the majority of chronic HBsAg carriers are sensitized to linear C‐gene‐derived structures in association with the host ‘self’ ‐component HSA.
Amino Acid Sequence Analyses of Non‐AA Proteins from Amyloid Fibrils of Bovine Kidney
Authors: VEIBY O. P. , SLETTEN K. , HUSBY G. , NORDSTOGA K.
Published: June 2006

The elution pattern obtained when amyloid fibrils from amyloid‐laden bovine kidneys were subjected to gel filtration under dissociating conditions revealed a larger amount of non‐AA material (eluting between the void volume and protein AA) than usually seen in other species. SDS PAOF, of this non‐AA fraction yielded several Coomassie blue stained bands. The most distinctive ones gave estimated molecular masses of 15 kDa, 18 kDa, 33 kDa and 43 kDa. These molecular species were electroblotted onto PVDK membranes, and wore further characterized by amino acid composition analyses, cyanogen bromide cleavage and N‐terminal analyses. The results revealed that the intermediate “non‐AA” fraction consisted of histones H2B, H3 and H4 in addition to protein AA also found in this fraction.