bluuebunny/crossref_metadata_2025 huggingface dataset:
The TcR‐CD3 Complex is Required for Activation of Human Lymphocytes with Staphylococcal Enterotoxin A
Authors: LANDO P. A. , DOHLSTEN M. , KALLAND T. , SJOGREN H. O. , CARLSSON R.
Published: June 2006

The involvement of the T‐cell receptor (TcR)‐CD3 complex in activation of human mononuclear cells by staphylococcal enterotoxin A (SEA) was investigated. TcR‐CD3 molecular complex expression was modulated with monoclonal anti‐CD3 antibodies. The proliferative response of such modulated cells to optimal doses of phytohaemagglutinin and monoclonal anti‐CD3 antibodies and to suboptimal doses of SEA was greatly impaired. In comcentrations above 1 ng/ml, SEA could also activate the modulated cells to a certain extent, apparently due to reexpression of low levels of the TcR‐CD3 molecular complex during incubation. TcR‐CD3 modulation decreased the ability of SEA‐activated cells to produce interleukin 2 and gamma interferon. Analysis of cloned cells revealed that SEA could only activate CD3+ but not CD3 clones. Both CD4+ and CD8+ clones were activated. The results show that sea activates human T cells by a mechanism involving the TcR‐CD3 molecular complex.
Increased Circulating HLA‐DR+CD4+ T Cells in Systemic Lupus Erythematosus: Alterations Associated with Prednisolone Therapy
Authors: RAZIUDDIN S. , NUR M. A. , AL‐WABEL A. A.
Published: June 2006

Patients with active systemic lupus erythematosus (SLE) in the circulation have a selective increase of a subset of the CD4+ helper/inducer T cells bearing HLA‐DR+, major histocompatibility complex class II antigens. We studied prednisolone‐induced alterations of HLA‐DR+, CD4+, and CD8+ T‐cell subsets in three patients with active SLE. Prednisolone therapy was accompanied by a drastic reduction in circulating HLA‐DR+, CD4+ T‐cell subsets, serum anti‐DNAtitre, normalization of the serum immunoglobulin profile, and CD4+ T‐cell responses to phytohaemagglutinin and concanavalin A. These changes in immune functions were associated with eventual improvement in the clinical condition of active SLE. A low precentage of HLA‐DR+, CD8+ T‐cell subsets was present in the circulation, which was not changed by prednisolone therapy. These results suggest that HLA‐DR+, CD4+ T‐cell subsets play a major role in the pathogenesis of active SLE, and that prednisolone‐induced immunosuppression in this disease is mediated by changes in the HLA‐DR+, CD4+ T‐cell subsets in circulating blood.
Arthritis in DBA/1 Mice Induced with Passively Transferred Type II Collagen Immune Serum
Authors: HOLMDAHL R , JANSSON L. , LARSSON A. , JONSSON R.
Published: June 2006

Arthritis was induced in DBA/1 mice by passive transfer of syngeneic anti‐type II collagen (CII) scrum concentrate. After transfer of serum containing 0.2 or 0.5 mg anti‐CII auto‐antibodies the first clinical signs of arthritis appeared 48 h after injection. Severe clinical arthritis was detected 96 h after injection. Immunohistochemical analyses of joints 48 h after serum injection revealed synovial foci in intercarpal and mclacarpophalangeal joints of macrophage‐like cells, expressing C3bi‐receptors and major hislocompatibility complex class II molecules, and infiltration of few CD4+ lymphocytes. Later (96 h afler injection), the inflamed synovia were dominated by C3bireceptor+ polymorphonuclear cells. In contrast to conventionally induced collagen arthritis (CIA), the inflammatory infiltrates, filling joint spaces and synovial tissue, were extensively dominated by polymorphonuclear cells, whereas macrophage‐like cells expressing class II molecules and a few T cells were seen only in the periphery of the developing pannus. The anti‐CII serum induced arthritis may be used as a model for studies of humoral mediated mechanisms operating in conventionally induced CIA as well as in rheumatoid arthrilis.
Blocking by Anti‐Idiotypic Antibodies of Monoclonal Antibody‐Mediated Protection Against Lethal Semliki Forest Virus in Mice
Authors: OOSTERLAKEN T. A. M. , HARMSEN M. , KRAAIJEVELD C. A. , SNIPPE H.
Published: June 2006

Semliki Forest virus‐(SFV) neutralizing monoclonal antibodies (MoAbs), produced after fusion of spleen cells from BALB/c mice and myeloma cell line P3‐X63‐AG8.653 or SP2/0, were used for anti‐idiotypic immunization of female BALB/c mice. Two intracutaneous immunizations (2 × 40 μg per animal), 3 weeks apart, with keyhole limpet haemocyanin‐conjugated MoAbs mixed with the saponin Quil A were sufficient to induce high levels of anti‐idiotypic antibodies in the circulation of these mice with the capacity to block specifically in vitro MoAb‐mediated virus neutralization. Anti‐idiotypic antibodies against SFV‐neutralizing MoAbs, either passively transferred or actively acquired by immunization, are also able to abrogate (specifically) passive immunity, mediated by critical protective doses of MoAb, in mice against infection with a lethal strain of SFV. Furthermore we confirmed by intervention with anti‐idiotypic serum in vivo that an SFV‐ neutralizing MoAb exerts its greatest protective effect during the first 2 days of infection.
apo‐SAA1/apo‐SAA2 Isotype Ratios during Casein‐ and Amyloid‐Enhancing‐Factor‐Induced Secondary Amyloidosis in A/J and C57BL/6J Mice
Authors: HÉBERT L. , GERVAIS F.
Published: June 2006

A/J mice are resistant while C57BL/6J are susceptible to casein‐induced secondary amyloidosis. One mechanism responsible for this phenotypic expression of resistance/susceptibiligy was shown to operate at the level of production of the ‘amyloid‐enhancing factor’ (AEF). AEF and processing of the apo‐SAA protein appear almost concomitantly during amyloidogenesis. In order to determine if AEF played a role in the processing of the apo‐SAA protein, three major parameters (apo‐SAA1/apo‐SAA2 ratios, level of AEF, and fibril formation) were determined during casein‐induced secondary amyloidosis. Kinetics of AEF production and serum levels of the two major apo‐SAA isotypes were compared in A/J and C57BL/6J animals. Both strains showed equal relative amounts of the two isotypes after seven, 15 and 21 casein injections, irrespective of the fact that the A/J strain had no detectable level of AEF and no amyloid deposition. An increased apo‐SAA 1/apo‐SAA2 ratio due to a decrease in apo‐SAA2 was noted after 38 days of casein injections when both strains had extensive deposits of amyloid fibrils. Involvement of AEF as an effector molecule was determined by following the ratio of the two major serum apo‐SAA isotypes and fibril formation during an accelerated protocol of amyloid induction in C57BL/6J animals. AEF had no direct effect on apo‐SAA isotype ratios in the serum.
Differential Activation of Monocytes in Haemodialysis Patients Exposed to Different Types of Membranes
Authors: BLUMENSTEIN M. , ZIEGLER‐HEITBROCK H. W. L. , SCHILLER B. , SCHMIDT B. , WARD R. A. , GURLAND H. J.
Published: June 2006

Peripheral blood mononuclear cells (PBMC) from haemodialysis patients treated with different types of membranes were isolated, incubated in vitro, and extracellular and cell‐associated interleukin 1 (IL‐1) assayed by radioimmunological methods. Extracellular IL‐1 was low and not different from controls, regardless of the dialyser used. In contrast, cell‐associated IL‐1 was increased in patients treated with dialysers containing low‐Hux Cuprophan (CU, n = 5) and polyacrylonitrile sheet membrane (AN69, n = 5), Patients treated with diaiysers containing highflux polysulphone (PS, n= 7), and polymethylmethacrylate (PMMA, n= S), exhibited no increase in cell‐associated IL‐l under these conditions. To elucidate the mechanism of the activation, aqueous extracts of dialysers containing CU, PS, and AN69 were tested for their ability to induce IL‐1 generation in PBMC from healthy donors. Extracts from unrinsed CU‐containing dialysers caused significant IL‐1 synthesis and release, whereas incubation with extracts from dialysers containing PS and AN69 sheet membranes did not. Hence, although both CU and AN69 sheet type dialysers result in activation of blood monocytes the mechanism of action appears to be different. We speculate that functional signs of PBMC activation as evidenced by increased spontaneous IL‐1 production observed in some patients on long‐term haemodialysis may result from extractable dialyser membrane material while in other instances direct cell membrane interactions or endotoxin transfer from the dialysate may be relevant.
Size of Jejunai Peyer's Patches and Migration of Lymphocyte Subsets in Pigs after Resection or Transposition ofthe Continuous Heal Peyer's Patch
Authors: ROTHKöTTER H. J. , ZIMMERMANN H. J. , PABST R.
Published: June 2006

In pigs there are two types of Peyer's patches in the small intestine; discrete patches in the jejunum (jejPP) and a continuous patch in the terminal ileum (ilPP). The ilPP was resectioned or transposed into the upper jejunum. After the operation the size of the remaining jejPP showed no compensatory growth in either group within 10 months. However, the number of CD8+ lymphocytes in the blood, spleen, mesenteric lymph nodes, tonsils, and Peyer's patches and the number of CD4+ cells in the spleen and tonsils was reduced in comparison to those of agematched control pigs. Autologous blood lymphocytes were labelled with fluorescein isothiocyanate and retransfused. In control animals the mid‐portion of the ilPP showed a lower entry of lymphocytes and the migration pattern of lymphocyte subsets was different in the animals with resectioned or transposed ilPP as compared to controls. Thus, the removal of the ilPP (about 60% of all small intestinal PP) did not result in the remaining patches adapting their size, but it did influence other lymphoid organs.
Bursin Localization in Mammalian Bone Marrow and Epithelial Cells of Intrahepatic Bile Ducts
Authors: AUDHYA T. , VIAMONTES G. , BABU U. , GOLDSTEIN G.
Published: June 2006

Bursin is a tripeptide (lysyl‐histidyl‐glycyl‐amide) found in follicular and dendritic reticular epithelial cells of the avian bursa of Fabricius that selectively induces the differentiation of committed B‐lymphocyte precursor cells but not of committed T‐lymphocyte precursor cells. We now show, in immunoassays with tissue extracts, that bursin is also present in avian and bovine bone marrow. There was, however, a categorical difference between avian liver (bursin‐negative) and bovine liver (bursin‐positive). Bursin was therefore isolated from bovine liver and bone marrow and the structure of mammalian bursin was determined; it was identical to avian bursin. Immunohistochemical examination of bovine liver showed the presence of bursin within epithelial cells of the intrahepatic bile ducts. These cells have previously been suspected of having an endocrine function because of the rich periductal capillary plexus, which coalesces to form a portal system draining into the liver sinusoids. These findings suggest that bone marrow is a site of bursin production and associated B‐cell differentiation in both birds and mammals. The bursincontaining cells of the intrahepatic bile ducts are not associated with developing B cells and it would appear that mammals have evolved a local hepatic function for bursin.
Expression of Activation Antigens on T Cells in Rheumatoid Arthritis Patients
Authors: POTOCNIK A. J. , KINNE R. , MENNINGER H. , ZACHER J. , EMMRICH F. , KROCZEK R. A.
Published: June 2006

The aim of our study was to identify differences in cell surface marker expression between T cells taken from the peripheral blood (PB) of healthy individuals and T cells recovered from inflamed joints of rheumatoid arthritis (RA) patients. Out of 118 monoclonal antibodies (MoAbs) directed against activation antigens on haematopoietic cells, 12 MoAbs recognizing nine distinct surface molecules were selected after a sereening procedure to study the expression of the corresponding antigens on T cells from the PB, synovial fluid and synovial tissue of RA patienls, and also on T cells from PB and spleens of controls. Using two‐colour flow cytometry and immunohistology we found the molecules B‐C5, CD39. CD40, CD45 R0, CD54, CD76 and potentially 1D11 to be substantially up‐regulated on T cells from various body compartments in RA patients. We thus could determine that the cell surface of T cells in RA patients not only differs in MHC class II expression, but also in a number of other activation‐associated cell surface molecules from T cells in healthy individuals.
Comparison of Biological and Immunological Activities of Human Monocyte‐Derived Interleukin 1β and Human Recombinant Interleukin 1β
Authors: MØLVIG J. , HANSEN B. SEHESTED , WORSAAE H. , HEJNAES K. R. , HELLE M. , DALBØGE H. , NERUP J.
Published: June 2006

Recombinant human interleukin 1β (rhIL‐1β) and supernatants of Escherichia coli lipopolysaccharides‐stimulated human monocyte (Mo) cultures, containing native human IL‐lβ (nhIL‐1β), demonstrate significant differences when tested in the mouse co‐stimulatory thymocyte (lymphocyte activating factor [LAF]) assay. The aims of the present study were to investigate this characteristic difference between rhIL‐1β and Mo culture supernatants (Mo supernatants), and to compare the biological and the immunological activity of preparations of rhlL‐lβ and nhlL‐1β during each step of an identical purification procedure. The biological activity of rhIL‐1β/nhIL‐lβ preparations was characterized by the use of the LAF assay and the rat islet insulin release assay. An IL‐1β enzyme‐linked immunosorbent assay (ELISA) was established in order to compare the biological and immunological responses of the IL‐lβ preparations.

We report that the significant difference between rhIL‐lβ and supernatants of Mo cultures, which was only demonstrable in the LAF assay, is due to the presence of interleukin 6 (IL‐6) in the Mo supernatants. We describe a simple cation exchange chromatography separating nhlL‐lβ and lL‐6 of Mo supernatants. The highly purified rhIL‐β possessing the correct amino‐terminal sequence and nhIL‐lβ have identical biological and immunological activities demonstrating a specific biological activity (SBA) of 3x102 U/ng IL‐lβ, Thus, we have no indications of secondary or tertiary structural differences between rhIL‐1β and purified nhIL‐lβ.

In contrast, both in the LAF assay and in the rat islet insulin release assay the SBA of anaminoextended rhIL‐lβ form, Met‐Glu‐Ala‐Glu‐rhIL‐lβ, was only 1‐2% of the SBA of rhIL‐lβ, suggesting that structural changes were introduced into the molecule by the amino‐terminal extension. In the present study we have demonstrated that systematic combined testing of IL‐lβ preparations in two different biological assays and an immunological assay is useful for the characterization and comparison of the activity of recombinant and native IL‐1β preparations purified by the use of exactly the same procedures.
Plasmodium berghei Subunit Vaccine: Repeat Synthetic Peptide of Circumsporozoite Protein Comprising T‐ and B‐Cell Epitopes Fails to Confer Immunity
Authors: MIGLIORINI P. , BOULANGER N. , BETSCHART B. , CORRADIN G.
Published: June 2006

In the murine malaria model induced by Plasmodium berghei, we studied the immunogenicity of the repeat region ofthe circumsporozoite (CS) protein, which is the main target of the antibody response in infected animals. We immunized several strains with a synthetic peptide‐‐Y(DPPPPNPN)3‐‐corresponding to one of the two P. berghei repeat sequences in complete Freund's adjuvant. Only C57BL/6 immunc sera reacted with the synthetic peptide in ELISA and with the native CS protein on P. berghei sporozoites, as detected by immunofluorescence. From lymph node cells of immunized C57BL/6 we isolated two repeat‐specific T‐cell lines which proliferated in the presence of the synthetic peptide or the recombinant CS protein. We analysed the protective role of this repeat‐specific response by injecting infectious sporozoites into mice immunized with irradiated sporozoites or with the repeat peptide. The percentage of mice developing parasitaemia was 80‐90% in the peptide‐immunized group and only 10‐20% in the group immunized with irradiated sporozoites. Anti‐repeat antibody titres were comparable in the two groups. On the basis of these results, we can conclude that the T‐ and B‐cell response to the CS repeat obtained with this synthetic peptide immunization is not sufficient for a protective immunity.
Lack of the G2m(n) Allotype in IgG Subclass Deficiency, in IgG2 Deficiency Together with Lack of G1m(a) and G3m(g), and in IgG3 Deficiency Together with Lack of G1m(f) and G3m(b)
Authors: OXELIUS V.A.
Published: June 2006

Lack of G2m(n) was demonstrated in both IgG2‐deficient and IgG3‐deficient Caucasian patients. Lack of G2m(n) or G2m(″,″) was found together with homozygosity for both Glm and G3m allotypes as the dominant finding, i.e. for IgG2‐deficient patients together with Glm (f,f) and G3m(b,b), constituting the Gm(f,″,b) phenotype, and for IgG3‐deficient patients together with Glm(a,a) and G3m(g,g), constituting the Gm(a,″,g) phenotype. The group with IgG2 deficiency and the selected patients with the Gm(f,″,b) phenotype expressed characteristically very low or undetectable IgG4, significantly increased IgG3, and normal IgGl. The group with IgG3 deficiency and the selected patients with the phenotype Gm(a,″,g) expressed instead normal IgG4 and nearly normal IgG2 and IgGl levels. The lack of G2m(n) together with lack of one or the other of the alternative Glm genes and corresponding G3m genes give different IgG2 levels and different IgG subclass patterns.

The frequency of Glm allotypes and corresponding G3m allotypes also deviated significantly when the IgG2 deficiency and IgG3 deficiency groups were compared with each other. Most IgG subclass‐deficient patients are homozygous in the Gm system and lack genetic variants in the three IgG subclasses, IgGl, IgG2, and IgG3.
Swainsonine, an Inhibitor of Glycoprotein Processing, Enhances Cytotoxicity of Large Granular Lymphocytes
Authors: YAGITA M. , SAKSELA E.
Published: June 2006

In the present study we investigated the effects of inhibitors of glycoprotein processing on cytotoxicity of human large granular lymphocytes (LGL). The incubation of LGL for 36 h with 0.5 μg/ml swainsonine (SW), which is an inhibitor of mannosidase II, resulted in the augmentation of cytotoxicity of LGL against an NK‐resistant colon carcinoma cell line (Colo‐320DM) without increase of binding frequency of LGL to target cells or of cell proliferation. The enhanced cytotoxicity was associated with increased binding of concanavalin A to SW‐treated LGL. The augmentation of cytotoxicity was also seen by 1‐deoxymannojirimycin (1‐DMN), an inhibitor of mannosidase I, but much higher amounts of this agent were needed to get the same level of augmentation as that with SW. Other inhibitors of glycoprotein processing such as castanospermine and 1‐deoxynojirimycin (1‐DN) did not show any augmentative effects on LGL cytotoxicity. The enhancement of cytotoxicity by SW was abolished by the addition of rabbit anti‐human interleukin (IL‐2) antibody to the culture. This result suggests that IL‐2 is involved in the augmentation of cytotoxicity of LGL by SW. The presence of SW in the culture of LGL together with IL‐2 also enhanced LAK generation compared to that with IL‐2 alone. Thus, our results suggest that SW should be recognized as an efficient immunopotentiator and that modulation of carbohydrate moieties elicited by SW may shed further light on the mechanism of LGL activation.
The Influence of FK‐506 and Low‐Concentration Ciclosporin on Human Lymphocyte Activation Antigen Expression and Blastogenesis: a Flow Cytometric Analysis
Authors: WOO J. , SEWELL H. F. , THOMSON A. W.
Published: June 2006

The influence of FK‐506 and ciclosporin (CsA), either alone or in combination, on PHA‐ and alloantigen‐induced human blood lymphocyte transformation was investigated. The concentrations of FK‐506 and CsA required to cause 50% inhibition of mitogen‐induced thymidine incorporation were 1.0 and 200 ng/ml respectively. Evidence of synergistic inhibition of DNA synthesis was obtained when doses of each drug below these concentrations were combined (FK‐506, ≤0.5 ng/ml; CsA ≤50 ng/ml). In contrast, FK‐506 and CsA failed to inhibit PHA‐induced increases in cell size and granularity determined by flow cytometric analysis at 1 and 3 days of culture. Moreover, no significant changes in the expression of the interleukin 2 receptor (IL‐2R; CD25), transferrin receptor (TR; CD7I) or HLA‐DR were observed in terms of percentage positive lymphocytes or mean cell surface membrane fluorescence intensity. In primary mixed lymphocyte cultures, 50% inhibition of thymidine incorporation was obtained with FK‐506 and CsA concentrations of approximately 0.25 and 25 ng/ml respectively. Evidence of synergy was obtained when lower doses of each drug were combined (FK‐506, ≤0.1 ng/ml; CsA ≤2 ng/ml). On their own, these concentrations of CsA were ineffective. In contrast to corresponding PHA‐stimulated cells, FK‐506‐treated alloactivated lymphocytes exhibited reductions in IL‐2R, TR and HLA‐DR antigen expression, which in the cases of IL‐2R and TR were further reduced by combination of FK‐506 with low‐concentration CsA. These data provide further evidence of the potential of combined low‐dosage FK‐506 and CsA for the control of human T‐cell activation and proliferation in response to allostimulation.
Mixed Enzyme‐Linked Immunosorbent Assay (MELISA) for HLA Class I Antigen: a Plasma Membrane Marker
Authors: BJERRUM O. W. , BORREGAARD N.
Published: June 2006

This study introduces a simple, reproducible assay for HLA class I antigen using antibodies against β2‐microglobulin and the heavy chain on HLA. The sandwich technique was named mixed enzyme‐linked immunosorbent assay (MELISA), and was designed for identification of plasma membranes in neutrophil subcellular fractions. The subcellular localization of HLA was identical to that of other plasma membrane markers, [3H]concanavalin A and detergent‐independent alkaline phosphatase, and was unchanged by stimulation of cells by weak and strong secretagogues. In addition to the presence as part of the HLA complex in the plasma membrane uncomplexed β2‐microglobulin is present in the specific granules of neutrophils. However, the release of β2‐microglobulin from intact neutrophils stimulated with formyl‐methionylleucyl‐phenylalanine was much higher than could be explained by exocytosis of specific granules. Subcellular fractionation studies demonstrated that β2‐microglobulin is localized in fractions characterized by latent alkaline phosphatase and released from this novel secretory compartment in response to stimulation with formyl‐methionylleucylphenylalanine.
Lymphocyte Subsets in the Blood
Authors: WESTERMANN J. , SCHWINZER R. , JECKER P. , PABST R.
Published: June 2006

Removal of the largest single lymphoid organ, the spleen, leads to an increase in severe infections. To prevent this, transplantation of splenic fragments can be performed, which may, however, cause an increase in CD8+ lymphocytes in the blood of these patients. This is controversial since in the clinical situation it is often difficult to account for the different age of the patients, the time point after the operation and many other factors known to influence the number of lymphocyte subsets.

Using a well‐defined animal model, B, T, CD4+, and CD8+ lymphocytes were determined preoperatively in adult rats. Then, either sham splenectomy, splenectomy, or splenic autotrans‐plantation was performed and the animals were followed up for 15 months after the operation.

The surgical procedure itself, the site of blood sampling and ageing all influenced the number of lymphocyte subsets profoundly. Furthermore, giving the data as relative or absolute numbers leads to different results.

Splenectomy caused lymphocytosis, due to a significant increase in B and CD8+ lymphocytes, as did splenic autotransplantation, which indicates that the number of lymphocyte subsets in the blood should not be used to argue in favour of or against splenic autotransplantation.

This study demonstrates that the number of lymphocyte subsets in the blood is influenced by many factors and therefore should be determined in a highly standardized fashion.
Ciclosporin‐Dependent, nu‐Independent, Mucosal Interleukin 6 Response to Gram‐Negative Bacteria
Authors: HEDGES S. , LINDER H. , DE MAN P. , EDÉN C. SVANBORG
Published: June 2006

Regulation of the mucosal inflammatory response to Gram‐negative bacteria was analysed. The interleukin 6 (IL‐6) secretion, influx of polymorphonuclear leucocytes into urine, and bacterial clearance from the kidneys were compared between Balb/c (nu/nu) and nu/± mice, with and without ciclosporin (CsA) treatment. There was no significant influence of the nu genotype on any of the host responses measured. CsA pretreatment significantly decreased II‐6 secretion in both nu/nu and nu/± mice, but did not affect bacterial clearance or the leucocyte response in any mouse strain tested. Tissue damage, in addition to bacterial infection, resulted in significantly higher levels of IL‐6 than bacterial infection alone. Tissue‐damaged mice were significantly less likely to clear the bacterial infection than their non‐damaged counterparts, but there was no significant difference in the leucocyte response. CsA pretreatment did not significantly reduce the levels of IL‐6 in the tissue‐damaged mice. These results demonstrate that the mucosal inflammatory response to Gram‐negative infection, including IL‐6 secretion, is nu‐independent, and that bacterial infection alone or in combination with tissue damage induce IL‐6 secretion by two different pathways.
Human Peripheral Blood‐Derived Dendritic Cells do not Produce Interleukin 1α, Interleukin 1β, or Interleukin 6
Authors: VAKKILA J. , SIHVOLA M. , HURME M.
Published: June 2006

Little is known about the non‐antigen‐specific signals delivered to T cells by dendritic cells (DC). Because several monocyte‐derived factors like interleukins 1α, 1β, and 6 (IL‐1α, IL‐1β, and IL‐6) enhance the T‐cell proliferative responses, we studied the production of the above‐mentioned cytokines by DC separated from human peripheral blood. The intracellular expression of the proteins (IL‐1α and IL‐6) was studied at a single‐cell level using an immunolabelling technique. The supernatants and cell lysates were studied with ELISA (IL‐1α and IL‐1β). Northern blotting analysis was used to quantitate the mRNA levels. Several approaches were taken to stimulate the production of IL‐1α, IL‐1β, and IL‐6 by DC. These included the incubation of the DC in the presence of either LPS, rIL‐1, or monoclonal anti‐HLA‐DR antibody, or the stimulation of cells with resting allogeneic T cells. None of the stimuli was able to induce the production of IL‐1α, 1L‐1β, or IL‐6 by DC, whereas LPS‐stimulated monocytes were strong producers of these mentioned cytokines and expressed the respective mRNA. Thus we concluded that IL‐1α, IL‐1β, and IL‐6 are primarily monocyte‐derived factors and that these factors are not needed or produced during the activation of resting T cells by DC.
Low T‐ and B‐Cell Reactivity is an Apparently Paradoxical Request for Murine Immunoprotection Against Streptococcus mutans
Authors: SOARES R. , FERREIRA P. , SANTAREM M. M. G. , DA SILVA M. TEIXEIRA , ARALA‐CHAVES M.
Published: June 2006

C57BL/6 mice thymectomized as adults or depleted of CD4+ cells were much less susceptible than intact conventional mice to the B‐cell mitogenic and specific immunosuppressive effects of a protein designated as F5′EP‐Sm secreted by Streptococcus mutans. These mice were also considerably more resistant to infection by these bacteria than intact individuals. The immunosuppressor effect of F5′EP‐Sm was also abrogated, however, in conventional intact mice when immunized intraperitoneally against heat‐inactivated F5′EP‐Sm. On the other hand, resistance to bacterial infection could be achieved by immunization of conventional intact C57BL/6 mice against heat‐inactivated F5′EP‐Sm by intraperitoneal or intradermal routes even when the animals were infected 3 months after immunization and even when the immunization procedure did not include Freund's adjuvant, which was the case with the intradermal route. Interestingly, the protection against the bacterial infection was accompanied by only a minor increase in specific serum antibodies against F5′EP‐Sm.

These results are discussed in the context of adequate strategies for immunoprotection against Streptococcus mutans and other micro‐organisms which are secretors of substances that share both B‐cell mitogenic and immunosuppressive properties and which are thus able to suppress the immune response by overstimulation of the immune system of the host.
Fine Specificity and Isotype Expression of Anti‐PC Antibodies in BALB/c, MRL/Mp‐1pr/1pr, and −+/+ Mice at Different Ages
Authors: GERBER V. E. , PONNAPPAN U. , HEUSSER C. H. , CINADER B. , BLASER K.
Published: June 2006

The MRL/Mp congenic mouse strains develop autoimmune disease with age. We have investigated age‐ and autoimmune‐related changes in fine specificity, isotype spectra and T15 idiotype expression of the anti‐phosphorylcholine (PC) response in BALB/c, MRL/Mp −+ and ‐lpr congenic mice and in (BALB/c x MRL/Mp‐lpr) F1 hybrids. Two groups of anti‐PC antibodies with distinct fine specificity are elicited in the memory response. Group I antibodies recognize the PC moiety and express the T15 idiotype. Antibodies of group II are specific for phenyl‐phosphorylcholine and are found predominantly in the memory response. In the MRL/Mp‐lpr and −+ strains only a minor population of antibodies expresses the T15 idiotype at all ages. However, a third group of antibodies was observed which binds to PC‐coated proteins and to Diplococcus pneumoniae R‐36A. This binding was not inibited by PC‐chloride and appeared mainly in the memory response at old age. The isotype distribution among anti‐PC antibodies was similar in all strains analysed. In the initial response primarily μ, γ3 and γ1 isotypes were produced, while in the memory response γ1 was dominant.

Thus autoimmune defects and ageing result in altered anti‐PC antibody and idiotype profile, probably related to altered states in both the T‐ and B‐cell compartments.
Zinc Ions Inhibit Factor I‐Mediated Release of CR1‐Bound Immune Complexes and Degradation of Cell‐Bound Complement Factors C3b and C4b
Authors: JEPSEN H. H. , TEISNER B. , SVEHAG S. E.
Published: June 2006

ZnCl2 exerted a dose‐dependent inhibition of citrate‐phosphate dextrose (CPD) plasma‐induced release of 125I‐labelled BSA‐anti‐BSA immune complexes (IC) bound to complement receptor lype I (CRl. CD35) in human whole blood. Maximal inhibition was observed at 10mM of ZnCl2.

Furthermore, the release of IC bound to erythrocyte (E)‐CRI by purified factor I. factor Ideficient serum plus purified factor I. or normal human. Serum was reduced by approximately 90%, 64%, and 52%, respectively, in the presence of 10 mM ZnCl2.

The effect of ZnCl2 on factor 1‐mediated degradation of cell‐bound C3b/C4b was also investigated employing CPD blood or E from a factor I‐deficient donor. These cells expressed covalently bound C3b and C4b as demonstrated by a simple agglutination technique. Upon incubation of CPD whole blood with purified factor I. or of E wilh purified factor I or normal CPD plasma, the C‐fragments were cleaved and the cells were no longer agglutinated by antibodies to C3c and C4c. The presence of ZnCl2 prevented Ihis factor 1‐mediaied degradation of C3b and C4b, as evidenced by the unaffected agglutination of the cells by the antibodies.

We conclude that ZnCl2 inhibited factor I aclivity since: (1) release of complementpreopsonized IC from E‐CR 1 by purified factor I was markedly inhibited (90%) in the presence of ZnCl2, (2) preincubation of the cells with ZnCl2 caused only a moderate inhibition (32 38%) of the IC release, and (3) degradation by purified factor I of covalently cell‐bound C3 band C4b was abrogated in the presence of 10 mM ZnCl2.
HLA Heterozygosity in Insulin‐Dependent Diabetes is Most Frequent at the DQ Locus
Authors: MICHELSEN B. , WASSMUTH R. , LUDVIGSSON J. , LERNMARK Å. , NEPOM G. T. , FISHER L.
Published: June 2006

Restriction fragment length polymorphism using an HLA‐DQ β‐chain genomic probe showed that 63 children with insulin‐dependent (type 1) diabetes mellitus (IDDM) were all (100%) positive for the BamH1 fragments 12 kb and/or 4 kb compared to 98% (62/63) for HLA‐DR3 and/or 4 and 75% (47/63) for HLA‐B8 and/or 15. The 36 (56%) DR3‐positive children were all 4‐kb‐positive; however, a total of 44 (70%) children were 4‐kb‐positive (P < 0.02). The 55 (87%) DR4‐positive children were all 12‐kb‐positive, but a total of 56 (89%) were 12‐kb‐positive (NS), The heterozygosity at the HLA region increased from 11/63 (18%) for HLA‐B8/15 to 29/63 (46%) for HLA‐DR3/4 (P < 0,0004) and to 37/63 (59%) for the HLA‐DQ 4 kb/12 kb fragments (P < 0.02). The test of an equal probability ofa positive result under the adjacent pair of tests indicates that the increased risk of developing IDDM in association with HLA‐DQ is to a great extent due to heterozygosity at this locus. There were no differences between the 4 kb/12 kb and the DR3/4‐positive IDDM children with respect to fasting or meal‐stimulated C peptide, insulin requirement, or levels of insulin antibodies formed during the first 12 months of insulin therapy. Our results support the hypothesis that HLA‐DQ is closely associated with an increased risk of childhood IDDM, and when typed for at this locus parameters of the clinical course were homogeneous, suggesting that factors other than HLA‐DQ may determine previously observed differences between IDDM children in clinical or functional parameters.
Thymus‐Independent B‐Cell Activation: Passively Incorporated Membrane Antibodies Serve to Focus the Antigen but Cannot Transmit Activation Signals
Authors: ALARCÓN‐RIQUELME M. , MÖLLER G.
Published: June 2006

We have activated B cells with membrane‐incorporated monoclonal antibodies against the hapten trinitrophenyl (TNP) using various concentrations of the polyclonal activator lipopolysaccharide (LPS) haptenated with TNP, We found that anti‐TNP‐decorated B cells were polyclonally activated by 1000‐fold lower concentrations of TNP LPS than untreated B cells or B cells decorated with antibodies of other specificities. In order to test whether the passively incorporated anti‐TNP monoclonals could mediate activation signals or only served to focus the polyclonal activator TNP‐LPS to the B cells, we compared the response of anti‐TNP‐decorated B cells from normal C3H/He mice and from the LPS‐unresponsive strain C3H/HeJ. We found that B cells from C3H/HeJ mice could not be activated to polyclonal antibody synthesis, in contrast to B cells from the LPS‐responsive strain C3H/He. Thus, contrary to what was previously suggested [9], the incorporated anti‐TNP antibodies could not mediate activation signals, but only served to passively bind and concentrate TNP‐LPS to the membrane of the B cells, thereby increasing the concentration of the polyclonal B‐cell activator LPS to the B cells.
CD26 Antigen is a Surface Dipeptidyl Peptidase IV (DPPIV) as Characterized by Monoclonal Antibodies Clone Til‐19‐4‐7 and 4EL1C7
Authors: ULMER A. J. , MATTERN T. , FELLER A. C. , HEYMANN E. , FLAD H.‐D.
Published: June 2006

In this study we investigated the binding or three different monoclonal antibodies (MoAb), TII 19‐4‐7,4EL1C7, and B1.19.2, which are clustered in CD26 to the ectoenzyme dipeptidyl peptidase IV (DPP IV) and to T lymphocytes. We found that all three MoAb bind to both unstimulated and mitogen‐stimulated T lymphocytes. Further results indicated an inconsistency within the CD26‐clusteredMoAb: TII 19‐4‐7 and 4EL1C7, but not Bl.19.2, recognized DPP IV on the surface of T lymphocytes and immobilized on solid‐phase ELISA or Western blot. There was compelition of binding to DPP IV between TII 19‐4‐7 and 4ELIC7. From these results we conclude that CD26 antigen is represented by the ectoenzyme DPP IV. TII 19‐4‐7 and 4ELIC7 recognize the same or partly identical epitopes on DPP IV, whereas B1.19.2 recognizes a different antigen. TI I 19‐4‐7 and 4ELlC7, but not B1.19.2, should be clustered in CD26.
Secretory Component‐Binding Properties of Normal Serum IgM
Authors: BOUVET J.‐P. , PILLOT J. , ISCAKI S.
Published: June 2006

Our aim was to investigate why serum IgM is poorly transferred into secretions in normal subjects. Indeed, the low IgM level in secretions contrasts with the eapacity of monoclonal IgM to bind to secretory component (SC), but it is not well established to what extent normal serum IgM can do so.

The mean SC affinity was studied with a polyclonal IgM preparation from 250 normal subjects and with a representative pool of 100 different monoclonal IgM. The SC‐binding percentages varied as a function of the IgM/SC molar ratio according to a common hyperbolic curve, with similar association constants: Ka= 4.19 ± 2.61 × 107 M‐1 (polyclonal pool) and Ka= 5.8 0 ± 2.73 × 107 (monoclonal pool). It thus appears that the large difference in IgM concentrations between blood and secretions cannot be due to an SC‐binding defect of serum IgM, but is probably explained by its low diffusion from blood to the extravascular compartment, compartment.
T‐Cell Recognition of HLA Class II Molecules Induced by Gamma‐Interferon on a Colonic Adenocarcinoma Cell Line (HT29)
Authors: LUNDIN K. E. A. , SOLLID L. M. , BOSNES V. , GAUDERNACK G. , THORSBY E.
Published: June 2006

HLA class II molecules may be induced on non‐lymphoid cells by gamma‐interferon (IFN‐y). We investigated if HLA class II molecules induced by IFN‐y on the HT29 colonic carcinoma cell line are functional, i.e, if they may be recognized by allogeneic T cells. We found that IFN‐y‐treated HT29 (HT29IFN) cells could not induce primary proliferative responses of peripheral blood T lymphocytes, nor were they able to induce proliferation in T‐lymphocytc clones (TLC) specific for HLA class II molecules found on HT29IFN. However, in the presence of exogenous interleukin 2 (lL‐2). 1 of 5 DQw8‐specific TLC proliferated when restimulated with HT29IFN, and 3 of these 5 TLC could very effectively inhibit the growth of HT29IFN, probably due to a cytoloxic effect. Both the proliferative response and the cytotoxicity were inhibited by anti‐DQ MoAb. We conclude that T cells may recognize HLA‐DQ molecules on non‐lymphoid cells, which may be of relevance for autoimmune diseases, graft‐versus‐host disease, and possibly for the recognition of malignant cells.
Acetylcholine Receptor Antibodies in Primary Biliary Cirrhosis: Characterization of Antigen and Idiotypic Specificity
Authors: SUNDEWALL A.C. , LEFVERT A. K.
Published: June 2006

Auto‐antibodies against the acetylcholine receptor of skeletal muscle are considered to be the cause of the neuromuscular dysfunction in myasthenia gravis. However, such auto‐anlibodies also occur in disease states not accompanied by neuromuscular symptoms. Patients with primary biliary cirrhosis have a high prevalence of different auto‐antibodies, including antibodies against the acetylcholine receptor. In primary biliary cirrhosis, these anti‐receptor antibodies are predominantly of IgM isotype. The IgM antibodies show a broader reactivity with receptors from other species than antibodies from myasthenic patients, Immunoglobulins from patients with primary biliary cirrhosis bear the same receptor antibody‐associated idiotypes, but ihe repertoire is quantitatively different from that found in myasthenia gravis patients. The IgM receptor antibody activity in a serum from a patient with primary biliary cirrhosis could be inhibited by cardiolipin, poly [dT], poly[I], and ssDN A, whereas this antibody activity in serum from a patient with myasthenia gravis was slightly reduced only by cardiolipin. Generally, IgM antibodies from patients with primary biliary cirrhosis had a broader reactivity with polynucleotides and phospholipids than IgG antibodies and antibodies from patients with myasthenia gravis. These results indicate a difference in the fine specificity between acetylcholine receptor antibodies in primary biliary cirrhosis and in myasthenia gravis.
CD4+, CD8+, and CD4− CD8− T Cells in CSF and Blood of Patients with Multiple Sclerosis and Tension Headache
Authors: MIX E. , OLSSON T. , CORREALE J. , KOSTULAS V. , LINK H.
Published: June 2006

Two‐colour flow cytometric analysis was performed on paired samples of peripheral blood (PB) and cerebrospinal fluid (CSF) of patients with untreated multiple sclerosis (MS) and, for reference, subjects with muscular tension headache (TH) using anti‐CD3, anti‐CD4, anti‐CD8, and anti‐HLA‐DR monoclonal antibodies in different combinations. CD4+/CD8+ T‐cell ratio was increased in CSF compared to PB in both MS patients and TH subjects to a similar extent. This was mainly due to higher CD4+ T‐cell levels in the CSF compartment. The proportion of HLA‐DR+ T cells was higher in CSF than PB in both MS and TH; this increase of DR+ T cells in CSF was more prominent in MS. The level of CD4+ CDS+ T cells, which represent a subset of activated T cells, was not different between CSF and PB, either in MS or in TH. The proportion of CD4− CD8− T cells, which were found generally not to be blast cells, was lower in CSF compared to PB in both patient groups. However, their CSF level was higher and their PB level lower in MS compared to TH. Results point to an accumulation of activated T‐helper cells in the CSF of both MS patients and healthy subjects. Fetal‐type CD4− CD8− T cells bearing the unusual T‐cell receptor γ/δ seem to be selectively recruited to the CSF of MS patients.
Quantitation of Antibody‐Secreting Cells in the Blood after Vaccination with aHaemophilus influenzaeType b Conjugate Vaccine
Authors: BARINGTON T. , HEILMANN C. , ANDERSEN V.
Published: June 2006

The human B‐lymphocyte response to proiein‐conjugated polysaccharide antigens has not previously been studied at the cellular level. In order to do so, we developed and evaluated haemolytic plaque‐forming cell assays detectingHaemophilus influenzaetype b (Hib) capsular polysaccharide‐specific antibody‐secreting cells (AbSC) of the isotypes IgM, IgG, and IgA. The appearance of AbSC in the blood after vaccimation of adults with diphtheria toxoid‐conjugated Hib polysaccharide was investigated. AbSC were detected from post‐vaccination day 5 to day 14.

IgA was the predominant isotype among these cells. IgM AbSC peaked slightly earlier (median day 7) than IgG and IgA AbSC (both day 8). On post‐vaccination day 8 ihe numbers of AbSC were: IgA. 1217/106mononuclear cells (median); IgG, 211; and IgM, 30 (n= 11). Similar isotype distribution has earlier been found after vaccination with pure capsular polysaccharides from Hib and pneumococci. The predominance of IgA AbSC in response to both conjugate and pure polysaccharide vaccines is probably due to reactivation of the same clones of IgA‐committed memory B cells originally primed at the mucosa by natural exposure to the polysaccharide or cross‐reacting antigens.
Cellular Fibronectin in Rheumatoid Synovium and Synovial Fluid: a Possible Factor Contributing to Lymphocytic Infiltration
Authors: WALLE T. K. , VARTIO T. , HELVE T. , VIRTANEN I. , KURKI P.
Published: June 2006

Mouse monoclonal antibodies against ED sequence‐containing cellular fibronectin (cFn) were used to show that Fn in the inflamed synovium is distinct from the major form of plasma Fn (pFn). An accumulation of cFn was seen at sites of hyperplasia of the rhcumatoid synovial membrane and in the walls of small vessels in the synovium by immunofluorescence microscopy. cFn was also found in rheumatoid synovial fluid by immunoblotting. Approximately one‐fifth of the T lymphocytes from rheumatoid synovial fluid bound to Fn, The binding of synovial fluid T cells was always higher than that from peripheral blood. These results have two implications. On the one hand, the cellular type of Fn may be an indicator of synovial inflammation. On the other hand, the deposition of Fn may be a factor contributing to the infiltration ol mononuclear cells into the synovium.
Role of Zinc in Interleukin 2 (IL‐2)‐Mediated T‐Cell Activation
Authors: TANAKA Y. , SHIOZAWA S. , MORIMOTO I. , FUJITA T.
Published: June 2006

In a serum‐free culture containing no zinc, zinc enhanced the proliferation of T cells in response to interleukin 2 (lL‐2), and also the in vitro production of lL‐2 by T cells. Although the lymphocyte proliferation was partially inhibited by anti‐IL‐2 antibodies, it was completely inhibited by anti‐IL‐2 receptor (CD25) antibodies. A Scatchard plot analysis showed that zinc induced the expression of high‐affinity receptors for IL‐2 on lymphocytes. The results indicated that zinc may be essentially required for IL‐2‐mediated T‐cell activation.
Immunological Typing of Acute Leukaemias by Rosetting with Immunomagnetic Beads: Comparison with Immunofluorescence Staining
Authors: SKJØNSBERG C. , BLOMHOFF H. KIIL , GAUDERNACK G. , FUNDERUD S. , BEISKE K. , SMELAND E. B.
Published: June 2006

Immunological phenotyping of acute leukaemias is important for a more precise diagnosis with respect lo both cell lineage and maturation level. We have developed a rapid and reliable method for immunophenotyping, based on the use of magnetic monodisperse beads coated with monoclonal antibodies. After only a 10‐min incubation of immunomagnetic beads (1MB) with mononuclear cells isolated from bone marrow or peripheral blood, the percentage of rosetting cells can be counted in the microscope. A panel of 16 monoclonal antibodies against haematopoietic cell‐surface antigens was applied on 29 cases of acute myclogenic (AML) or lymphocytic (ALL) leukaemias, in order to compare immunological typing by immunomagnetic beads with immunofluorescence staining (IF). In all the cases tested, the two methods showed a virtually identical antigen distribution. The procedure described offers the advantages of being fast and simple to perform. Moreover, it has a high specificity and is easy to interpret in cases with low antigen expression.
Impact of 90Sr on Mouse Natural Killer Cells and their Regulation by Alpha Interferon and Interleukin 2
Authors: GIDLUND M. , BIERKE P. , ÖRN A. , AXBERG I. , RAMSTEDT U. , WIGZELL H.
Published: June 2006

Male CBA/SU mice were exposed to ionizing radiation by intraperitoneal injection of the boneseekiag β‐emitter 90Sr, NK‐cell lytic activities in spleen, peripheral blood, and lymph nodes were severely depressed or completely abolished. In contrast, production of the NK regulatory proteins alpha interferon (IFN‐x) and interleukin 2 (IL‐2) was normal 5–8 weeks after 90Sr injection. IFN‐x, produced in vivo or in vitro by cells from injected mice, was able to enhance strongly NK lytic activities. These data indicate that 90Sr acts on the bone marrow, where it interferes with ihe maturation and seeding of NK precursor cells. The mechanisms regulating NK activities in peripheral organs remained relatively unchanged. Finally, we did not detect any major organ redistribution of NK cells as a result of 90Sr irradiation.
A Segment of Human Vh Gene Locus is Duplicated
Authors: CHEN P. P. , YANG P.M.
Published: June 2006

In humans, the kappa light chain variable region gene (Vk) locus evolved in part by duplications of four large segments. Similarly, some portions of the human heavy chain constant region gene locus are duplicated. Recently, we found that the Humhv3005 Vh gene is highly homologous to the reported 1,9111 gene. Subsequently, restriction fragment length polymorphism study of the human Vh locus with a 1.6‐kb EcoR1 fragment downstream of the hv3005 gene (termed hv3005/E1.6) suggested that the hv3005 and the 1.9III Vh loci might be generated by duplication from a common‐ancestor Vh gene segment. To assess this possibility, we mapped the 15‐kb region of the isolated hv3005 clone, beginning from 2 kb upstream, and sequenccd the adjacent Vh4 gone (designated Humhv4005) located 10 kb downstream of the hv3005 gene. The result showed that hv4005 shared 99% homology with the 1.9II gene, located about 11 kb downstream of the 1.9III gene. Taken together, these data demonstrate that the hv3005‐hv4005 region and the 1.9III‐1.9II region arose by a duplication of a common ancestor Vh gene segment.