bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Natural Killer Clones Enhance in Vitro Antibody Production by Tumour Necrosis Factor Alpha and Gamma Interferon
Authors: BECKER J. C. , KOLANUS W. , LONNEMANN C. , SCHMIDT R. E.
Published: June 2006

To determine whether NK cells are involved in the regulation of the antibody response, we studied the effects of human NK clones on B‐cell growth and differentiation and the mechanisms involved. We demonstrate that various human NK clones enhance the immunoglobulin production of SAC/rlL‐2‐activated B cells, e.g. IgG and IgM by up to 230% and anti‐tetanus toxoid antibodies by up to 430%. Cell‐cell interactions via cell‐surface structures, e.g. the CD11a/CD18 molecule, were found to be critical. Subsequently the NK‐mediated B‐cell regulation involves cytokines, since cell‐free supernatants obtained by 48‐h cultures of NK clones exerted BCGF and BCDF activity. Neutralization studies and direct determination characterized these cytokines as IFN‐γ and TNF‐α. The cytokine production of NK clones could be triggered by activated B cells only. Northern blot analysis demonstrated that activated B cells in co‐culture with NK clones were able to induce accumulation of mRNA transcripts for IFN‐γ and TNF‐α in NK cells.
Epitope‐Specific Regulation and the Literature
Authors: ELLIOTT J. I.
Published: June 2006

The antibody response to a hapten conjugated to a protein can, in a number of situations, be suppressed by prior immunization with the carrier protein alone. The study of this finding appears to have been handicapped by previous literature on the subject having been consistently misquoted. It is to be hoped that recognition of this will allow a better understanding of the limitations to such suppression, and hence the mechanism by which it may act.
Anti‐DNA Antibodies Induced by BK Virus Inoculations
Authors: FREDRIKSEN K. , TRAAVIK T. , REKVIG O. P.
Published: June 2006

The ability of BK virus to induce anti‐DNA antibodies in rabbits, and the ability of these antibodies to bind natural eukaryotic DNA and synthetic polynucleotides have been analysed. The specificity of the binding was assayed by inhibition of anti‐dsDNA and ‐ssDNA ELISA tests with dsDNA, ssDNA, and synthetic single‐stranded as well as double‐stranded polynucleotides The anti‐dsDNA activity of two rabbit antisera was effectively inhibited by dsDNA and ssDNA and poly(dAdT)‐poly(dAdT). The other nucleotide antigens produced relatively loss inhibition. The anti‐ssDNA binding was most efficiently inhibited by the homologous antigen, whereas inhibition by dsDNA only reached approximately 70%, of the maximum as defined by ssDNA as inhibitor. This indicates the existence of a selective anti‐ssDNA antibody population and, a population recognizing both ssDNA and dsDNA within the sera. Cross‐reaction of the induced anti‐DNA antibodies with phospholipid antigens, such as cardiolipin, phosphatidylic acid, and bacterial cell surface, could not be demonstrated We conclude that antibodies resulting from inoculation with BK virus specifically bind to dsDNA and ssDNA and possess a high affinity for the synthetic duplex poly (dAdT). In this way. they have some similarities with anti‐DNA antibodies encountered in SLE (systemic lupus erythematosus) in both man and mouse,
Carrier Effect of Concanavalin A‐Reactive and ‐Non‐Reactive Material in Tuberculin PPD
Authors: HARBOE M. , WIKER H. G. , LACHMANN P. J.
Published: June 2006

Tuberculin purified protein derivative (PPD) is a very potent T‐cell reactive material in tuberculin‐positive individuals, but the components responsible for this reactivity have not been adequately defined. Three purified mycobacterial proteins (MPB70. the BCG 65‐kDa protein, and BCG antigen 85B) with different degrees of temperature sensitivity were iodine‐labelled and added to BCG culture fluid, and the mixtures were autoclaved at 120° C for 30 min to simulate initial heating procedure used to prepare PPD. SDS PAGE Followed by protein staining and autoradiography showed that the banded pattern of unhealed culture fluid was completely lost after heating, and only the labelled MPB70 preparation retained most of the radioactivity in a fraction with soluble protein of the same size. Most mycobacterial proteins are extensively denatured by these procedures, which explains the previous extensive difficulties in isolating defined constituents from PPD to characterize their behaviour in B‐ and T‐cell reactions. In assays for the carrier effect of NIP‐PPD for induction of anti‐NIP production in BCG‐vaccinated mice, the active fractions were heterogeneous in lectin reactivity as well as in SDS PAGH. It appears most likely that a number of Mycobacterium tuberculosis proteins give rise to core peptides which resist proteolysis and heat denaturation to possess powerful T‐cell‐activating ability.
Phenotypic and Functional Changes of Tumour Cells from Patients Treated with Monoclonal Anti‐Idiotypic Antibodies
Authors: ALLEBES W. A. , KNOPS R. , BONTROP R. E. , OTTING N. , RAEMAKERS J. , HAANEN C. , CAPEL P. J. A.
Published: June 2006

In this paper data are presented indicating that immunotherapy with monoclonal anti‐idiotypic antibodies (MoAb anti‐id) can provoke different responses in the B‐cell tumour concerned. With respect to the course of disease during and after immunotherapy, the in vitro findings may very well explain the in vivo observations in the two patients (D.E.F. B.O.R.) with B‐cell chronic lymphocytic leukaemia (B‐CLL) who were treated with MoAb anti‐id. After initial tumour reduction, there was a recurrence of tumour cells with altered functional and phenotypic properties. In both cases the recurring tumour cells still expressed the same idiotype.

In one patient (D.E.F.) the phenotypic changes (a surface Ig change from IgM, IgG, IgA, and IgD to weakly positive IgM and IgD) and functional changes (a 10‐fold increase in [3H]thymidine uptake and a decreased idiotype secretion in vitro), together with the in vivo findings with respect to the course of disease–at relapse an impressive tumour regrowth rate with constant serum idiotype level–suggest that immunoselection might have taken place favouring the survival and relapse of a less mature, more aggressive tumour cell population with a lower idiotype expression.

In the second patient (B.O.R.), the phenotypic changes (an isotype change from IgM and IgD to IgM with the loss of IgD, and a gradual decrease in expression of CD 19 and CD24) and functional changes (a 10‐fold increase of idiotype secretion in vitro), together with the in vivo finding that the serum idiotype level had increased 25‐fold compared with the preimmunotherapy serum level with comparable tumour load, strongly suggest an immunotherapy‐induced differentiation of the malignant B cell.

We also describe an increased expression of CD74, detected by MoAb BoM22, on the recurring tumour cells of patient B.O.R. whereas the expression of HLA‐DP, ‐DQ and ‐DR did not change. The significance of this finding is unclear.
Production of 15‐Hydroxyeicosatetraenoic Acid by Purified Human Eosinophils and Neutrophils
Authors: MORITA E. , SCHRÖDER J.‐M. , CHRISTOPHERS E.
Published: June 2006

In the presence of high concentrations of exogenous arachidonic acid (≥ 10 μm). eosinophils produced 15‐hydroxycicosatetraenoic acid (15‐HETE) in the absence of stimuli. The calcium ionophore A23187, as well as the chemotaxins used in this study‐complement split product C5a, platelet‐activating factor (PAF). and,N‐formyl‐methionyl‐leucyl‐phenylalanine(FMLP)–failed to increase 15‐HETE production, indicating that eosinophil 15‐lipoxygenase is already active Production of 15‐HETE from eosinophils increased with increasing concentrations of arachidonic acid, exogenously added. Maximal 15‐HETE production was observed to he 1111 ± 380 ng per 106 eosinophils at the concentration of 100μm of arachidonic acid. With low concentrations of exogenous arachidonic acid (below 2;μm). eosinophils were considered to incorporate exogenous arachidonic acid into their cell membrane, and did not produce 15‐HETE. In contrast, 15‐HETE formation in highly purified neutrophils (eosinophils < 1%) was negligible compared with that in eosinophils (.300‐fold less), suggesting that 15 f HETE‐forming activity in granulocytes is derived from the eosinophil 15‐lipoxygetiase pathway and that neutrophils may lack 15‐lipoxygcnase activity.
Synovial Cells Responding to a 65‐kDa Mycobacterial Heat Shock Protein have a High Proportion of a TcRγδ Subtype Uncommon in Peripheral Blood
Authors: SöDERSTRÖM K. , HALAPI E. , NILSSON E. , GRÖNBERG A. , VAN EMBDEN J. , KLARESKOG L. , KIESSLING R.
Published: June 2006

We have analysed the ability of T cells from synovial fluid mononuclear cells (SFMC) and from peripheral blood mononuclear cells (PBMC) of inflammatory arthritic diseases to proliferate in response to mycobacterial antigens (65‐kDa heal shock protein [hsp] of BCG. whole BCG) and to rat collagen type II. The SFMC demonstrated a significantly greater ability to respond to 65‐kDa hsp of BCG. and to whole BCG, compared with PBMC from the same patients, With collagen type II, only a small proportion of the patients showed a proliferative response, although with this antigen also SFMC responded better than PBMC. There was no difference between SFMC and PBMC in the response to control antigen (tetanus toxoid), phytohaemagglutinin (PHA), or interleukin 2 (IL‐2). A high proportion of cells in SFMC‐derived short‐term T‐cell lines were of TcRγδ type, often exceeding the number of TcRγδ type. There was a significantly higher proportion of TcRγδ cells in the SFMC lines compared with the PBMC lines, and a large part of the TcRγδ cells in she SFMC cultures was CD8+ The SFMC lines had a high proportion of δ‐ TCS‐1+ cells (Vδ1) among their TcRγδ cells, always exceeding the percentages of TiγA+ (Vγ9) and BB3+ (Vδ2) in the PBMC lines, the distribution of TcRγδ subtypes was markedly different, with a TiγA+/BB3+ population in the majority. These data argue for a different subpopulation distribution of TcRγδ cells in synovial fluid compared with peripheral blood of patients with inflammatory arthritic diseases.
Site of Catabolism of Autologous and Heterologous IgA in Non‐Human Primates
Authors: MOLDOVEANU Z. , MORO I. , RADL J. , THORPE S. R. , KOMIYAMA K. , MESTECKY J.
Published: June 2006

Because of similarities between the human and monkey immune systems, we considered the monkey a suitable model for studies on the catabolism of various molecular forms of IgA, for which little information is available. The residualizing label dilactilol‐[125I]tyramine was coupled to monkey (Macaca fuscata) IgA and IgG, as well as to human monomeric and polymeric myeloma IgA1 and IgA2 proteins. When labelled proteins were injected intravenously into monkeys, the non‐metabolizable radioiodinated tracer accumulated at the cellular site of protein degradation, allowing identification of the catabolic sites. To determine the uptake or injected proteins by various tissues, monkeys were sacrificed 6‐7 days after injection of labelled proteins, when blood‐associated radioactivity was ≥ 10% of the injected dose, as measured by plasma clearance. When monkey or human monomeric IgA. as well as human polymeric IgA, irrespective of subclass, was administered to monkeys, the liver showed the greatest tissue uptake relative to total dose injected and to organ weight, and the highest acid soluble radioactivity (degraded protein). Although both hepatocytes and non‐parenchymal liver cells were involved in IgA uptake, the hepatocytes were more active. Therefore, it appears that the liver is the major site of uptake and catabolism of IgA in monkeys and possibly in humans.
The MHC Class I Associated Beta2‐microglobulin (β2m) Light Chain is Expressed in a Molar Excess Over HLA‐ABC and CD1 on the Membrane of Leukaemic B Cells but not Leukaemic T Cells: Evidence for Further β2m‐Associated Molecules
Authors: JONES R. A. , CHILD J. A. , MASTER P. S. , SCOTT C. S.
Published: June 2006

Beta2‐microglobulin (β2m) constitutes the common light chain of both the MHC‐encoded HLA‐ABC molecules and a group of structurally related glycoproteins recognized by antibodies of the first cluster of differentiation (CD1a, CD1b and CD1c). These CD1 antigens appear similar to murine T1 and Qa molecules in terms of structure and tissue distribution, although the question of inter‐species homology is controversial. A further group of alloantigens expressed predominantly on T cells has been reported however, with immunogenetic characteristics more closely analogous to the murine T1/Qa system than the CD1 antigens, although their precise identity remains ill‐defined. Having previously shown that malignant B cells may express membrane CD1c, we examined leukaemic B‐cells corresponding to early lymphoblastic differentiation (null‐and common acute lymphoblastic leukaemia) through to the terminal plasma cell stage for the expression of other non‐HLA class I β2m ‐associated molecules. It was found that leukaemic B‐cells at intermediate/late stages of differentiation, represented by non‐Hodgkin's lymphoma (B‐NHL) and‘hairy‐cell’leukaemia (HCL), had significantly higher β2m: HLA‐ABC ratios than did the cells from other types of B‐cell malignancy. Although leukaemic B cells with a demonstrable non HLA‐ABC‐ associated β2m component expressed detectable levels of CD1c. and insignificant levels of CD 1a and CD 1b, the antigen density was insufficient to account for the excess β2m. In vitro stimulation of leukaemic B cells by phorbol ester substantially increased the expression of HLA‐ABC and CD1c, but also accentuated further the difference between the expression of these molecules and that of β2m. There was no detectable β2m other than that associated with HLA‐ABC and CD1 on the surface of malignant T cells by contrast. Our findings strongly support the existence, at certain stages of leukaemic B‐cell differentiation, of an additional β2m component(s) other than that associated with HLA‐ABC and CD1.
Glycosaminoglycans in Extracts of Cardiac Amyloid Fibrils from Familial Amyloid Cardiomyopathy of Danish Origin Related to Variant Transthyretin Met 111
Authors: MAGNUS J. H. , STENSTAD T. , KOLSET S. O. , HUSBY G.
Published: June 2006

We have previously demonstrated an association between secondary AA type amyloid fibrils and glycosaminoglycans (GAGs) in human liver. The present study was aimed ill investigating whether a similar association could be demonstrated in isolated cardiac amyloid fibrils from a unique Danish family with amyloid cardiomyopathy related to variant transthyretin (TTR) with a single amino acid substitution of a methionin for leucine at position 111 (TTR Met 111).

Using gel nitration and ion exchange chromatography. significant amounts of GAGs were detected in close association with purified myocardial amyloid fibrils, whereas only truce amounts of polysaecharides were present in the corresponding normal preparation. The GAGs were identified as 50% chondroitin sulfate, 33% heparin/heparan sulfate, and 17% hyaluronan. With the methods used the amyloid associated GAGs appeared as high molecular weight free polysaccharidc chains, and not as part of intact proteoglycans (PGs) in the fibril extracts.

We conclude that the association between purified amyloid fibrils and GAC is may be a general feature of amyloid deposits. Also, we suggest that the proportion of different GAGs in the amyloid deposits may depend both on the organ or tissues affected and the type of protein making up the fibrils.
Monocyte Chemotactic Activity in Sera after Hypnotically Induced Emotional States
Authors: ZACHARIAE R. , BJERRING P. , ZACHARIAE C. , ARENDT‐NIELSEN L. , NIELSEN T. , ELDRUP E. , LARSEN C. SCHADE , GOTLIEBSEN K.
Published: June 2006

In a number of studies it has been shown that psychological factors in general and specifically emotional factors can be correlated to changes in immunological function and defence mechanisms. Although the mediating pathways between the central nervous system and the immune system still remain unclear, it is known that some of the‘classical stress hormones’such as cortisol and catecholamines have modulatory effects on different immunological parameters. In this investigation we wished to study the effect of brief hypnotically induced emotional States on monocyte chemotaxis and endocrinological parameters. Eleven highly hypnotically susceptible volunteers were, while in a deep trance, given suggestions to re‐experience earlier life experiences involving intense anger and depression in random order. Before concluding hypnosis subjects were given suggestions to re‐experience a feeling of happiness and well‐being. Monocyte chemotactic activity in sera and serum levels of cortisol, as well as venous plasma levels of the catecholamines epinephrine, norepinephrine. DOPA and DOPAC. were measured before hypnosis, after each emotional stale and immediately after hypnosis. The results showed a significant difference (P<0.02) in chemotactic activity between the angry and the depressed emotional states, the depressed state exhibiting a decreased chemotactic index compared with the angry slate. Chemolactic index after the happy relaxed emotional state also showed a significant (P<0.01) increase compared with both chemotactic index before hypnosis and chemotactic activity after the angry and depressed state. Though there were significant differences between emotions and between emotions and the before‐hypnosis‐concision, no clear‐cut significant differences between the emotional slates of anger and depression could be detected for serum cortisol levels and catecholamine plasma levels. Significant positive correlations (P<0.01) for differences in chemotactic activity and differences in plasma DOPA levels between emotional states was found. When investigated in vitro, DOPA did not in itself exhibit monocyte chemotactic properties. No other significant correlations between differences in chemotactic activity and other endocrinological parameters could be detected. Soluble interleukin‐2 receptors in serum were also measured. No significant differences were found.
Detection, Isolation and Functional Studies of CD25+ T Cells in Lymph Nodes Involved by B‐Cell Non‐Hodgkin's Lymphomas
Authors: BONNEFOIX T. , CLARET E. , PICCINNI M.‐P. , JACOB M.‐C. , ZHENG X. , SOTTO J.‐J.
Published: June 2006

We searched for the presence of IL2 receptor (CD25) on T cells as an activation marker m lymph nudes involved by B‐cell non‐Hodgkin's lymphomas (B‐NHL). In 26 malignant lymph nodes studied, the number of CD25+ T cells among total T cells was usually low when assessed by immunofluorescence analysis (mean ± SD: 6.7%± 11.2%), but greatly increased when an immunomagnetic rosette method was used (mean±SD: 17.5% plusmn; 16.6%). In six cases. CD25‐/CD25/CD25+ cells were isolated by immunomagnetic separation, with a purity greater than 97%, for both populations. Expansion of CD25‐/CD25+ T cells was obtained with IL2 and PHA, then conditioned media (CM) were prepared. No IL2 activity was found in CM from both CD25‐/CD25+ T cells when tested on CTLL2 cells. BCGF and BCDF μ/γ activities were assayed on normal B cells stimulated with soluble or insolubilized anti‐μ antibodies (BCGF) or with Cowan I (BCDF). Results of production of all these activities were comparable for both populations, and thus do not favour the possibility that CD25+ T cells closely associated with malignant B‐NHL cells in lymph nodes may influence their proliferation (BCGF) or expression/secretion of heavy chain isotype (BCDF μ/γ).
Loss of HLA Heavy Chain and β2‐Microglobulin in HLA Negative Tumours
Authors: CABRERA T. , CONCHA A. , RUIZ‐CABELLO F. , GARRIDO F.
Published: June 2006

Expression of HLA class I antigens on the cell surface requires the association of heavy chain. endogenous peptide and β2‐microglobulin (β2m). We have studied the expression of free class I heavy chain and β2m in cryostatic sections of 51 HLA class I negative human tumours of different origin. Selective low expression of β2m was observed only on the 9 colorectal adenocarcinomas. In contrast, simultaneous under‐expression of heavy chain and β2m was detected in the rest of the tumours studied. Our results suggest that the accumulation in the cytoplasm of HLA heavy chain in HLA negative colorectal carcinomas is a particular mechanism that cannot be applied lo all HLA negative tumours.
Acute Phase Protein, Serum Amyloid A, Inhibits IL‐1‐ and TNF‐Induced Fever and Hypothalamic PGE2 in Mice
Authors: SHAINKIN‐KESTENBAUM R. , BERLYNE G. , ZIMLICHMAN S. , SORIN H. R. , NYSKA M. , DANON A.
Published: June 2006

The effect of serum amyloid A(SAA)on fever induced by recombinant interleukin‐1b(ril‐1β) or recombinant tumour necrosis factor α (rTNFα) was studied in mice. Serum amyloid A is an acute phase protein whose rise in pathological events is induced by the cytokines IL‐1. IL‐6 and TNF. Administration of human serum amyloid A to mice inhibited fever induced by rlL‐1β or TNFα in vivo, while the addition of human serum amyloid A lo mice hypothalamic slices inhibited IL‐1β or TNPα‐induced prostaglandin E2 (PGE2) production in vitro. Since serum amyloid A did not affect body temperature or hypothalmic PGE2 levels when administered alone, it may represent a specific servo‐mechanism for fever regulation in acute events, and it suggests, for the first time, a possible feedback relationship between serum amyloid A and the immunoregulatory cytokines.
The Human in Vitro Anti‐type 4 Pneumococcal Polysaccharide Antibody Response is Regulated by Suppressor T Cells
Authors: GRIFFIOEN A. W. , RIJKERS G. T. , TOEBES E. A. H. , ZEGERS B. J. M.
Published: June 2006

In order to study the regulation of the human B‐cell response to T‐cells independent type 2 (TI‐2) antigens, we analysed the role or T‐cells in the in vitro antibody response to type 4 pneumococcal polysaccharide (PS4). We found that T cells can positively regulate the in vitro antibody response to PS4 when they arc added into an in vitro B‐ceil culture system. In addition we demonstrated that T cells exert a negatively regulating activity. We found that T cells, when cultured for 24 h in the presence of high concentrations of PS4, can suppress the anti‐PS4 antibody response. This down‐regulation of the anti‐PS4 B‐cell response is shown to be antigen specific and MHC‐restricted. Furthermore, PS4‐specific T‐cell mediated suppression appears lo be radioresistant and confined to the T‐ceil preparations enriched for CD8+ cells. The results show that analogous to results obtained in mice, human T cells are able to exert a regulatory control of the antibody response to Ti‐2 antigens.
High Plasma Fibronectin Levels in Multiple Myeloma Patients: Possible Mechanisms and Clinical Implications
Authors: PAIZI M. , MANASTER J. , QUITT M. , SPIRA G.
Published: June 2006

Fibronectin (FN) has been implicated in the formation cryoprecipitates in rheumatic diseases and is present in tissues where, under pathological conditions, immune complexes are frequently deposited. We found elevated levels of FN in the plasma of 92% of multiple myeloma patients tested compared with a group of normal subjects, although the level of FN did not correlate with the level of the paraprotein. We then characterized the interacting fragments on both molecules and found that under physiological conditions of pH and ionic strength both heavy and light chain of all multiple myeloma and normal IgG showed affinity to FN: the active fragment on FN was found to be the aminoterminal heparin‐binding domain. These findings raise the possibility that FN might be implicated in some of the clinical symptoms of multiple myeloma.
Inhibition of Neutrophil Chemotaxis by Elastase‐Generated IgG Fragments
Authors: ECKLE I. , KOLB G. , HAVEMANN K.
Published: June 2006

IgG1 is cleaved in vitro by granulocyte elastase into Fc and Fab fragments. The elastase‐specific Fe fragment has been previously detected in vivo. Biological activity of the fragments has been described in modulating neutrophil oxidative metabolism and enzyme release. To investigate further effects granulocyte chemotaxis(CT) was tested. The CT was assayed in Boyden chambers and the chemotactic index (CI) was calculated which represents the mean distance travelled by the activated cells.

Stimulation of leucocyte CT by casein, activated scrum and FMLP gives maximal values of Δ CI = 46.7, 26.4 and 7.1 μm, respectively. Native IgG1 and the elastase‐produced IgG fragments do not stimulate leucocyte CT.

FMLP‐stimulated CT is specifically inhibited by the elastase‐produced Fc fragments. Addition of 7 nmol Fc to stimulus concentrations of 16 to 125 nM FMLP results in total inhibition of chemotaxis demonstrated by CI values which are lower than those for unstitmulated cells. The inhibition of CT is concentration dependent in the range of 2 to 7 nmol Fc/106PMN.

Number and affinity of FMLP receptors are not incipience by Fc fragments, so Fc binds neither to FMLP nor the FMLP receptor.

CT stimulated by casein shows a large portion of chemokinesis. Only at suboptimal casein concentrations do Fc and IgG have an Inhibitory effect on CT (0.63 mg casein /ml. 10 nmol peptide/106 PMN).

C5a‐stimulaicd CT is not influenced by IgG or IgG fragments which indicates that the samples are not cytotoxic. So the FMLP and casein‐stimulated CT is specifically inhibited by the elastase‐produced Fc fragments in a low concentration range.
Idiotypic Replica of an Anti‐Human Tumour‐Associated Antigen Monoclonal Antibody DNA Sequence Comparison between Ab1 and Ab3
Authors: FLAMINI G. , MARCHE P. N. , CAZENAVE P. A. , NATALI P. G. , SICCARDI A. G. , VIALE G.
Published: June 2006

2G‐3 is an anti‐anti‐idiotypic MoAb (Ab3) obtained upon immunization wiih a monoclonal Ab2 (A3B10), which behaves as the “internal image” of CaMBrl. a saccharidic epitope defined by MoAb MBrl (Abl), CaMBrI is expressed on glycoconjugates of the human mammary carcinoma cell line MCF‐7 und on normal and neoplastic mammary gland epithelial cells, Abl and Ab3. although exhibiting, in many respects, superimposable paratopic and idiotopic specificities, show a non‐identical fine immunoreactivity, since 2G‐3 has a preferential reactivity with the saccharidic epitope mounted on glycoproteins. while MBrl reacts with both glycoproteins and glycolipids, V‐region sequence analysis has shown that: (i) the VK genes employed belong to different families(VK 1 and VK 10); (ii) the J K2 segment is shared by the two L chains(thus a high degree of homology is observed between VK CDR3s): (iii) the VH genes employed derive from the same family VHIIBJ55K (but show CDR homology only in CDR2): (iv) different JM region genes are employed. These data together with the comparison of deduced secondary structure parameters give further evidence fur the possible production of similar combining sites using different VH and VL germ‐line genes.
Induction, Specificity and Elimination of Asialo‐GM 1+ Graft‐Versus‐Host Effector Cells of Donor Origin
Authors: GHAYUR T. , XENOCOSTAS A. , SEEMAYER T. A. , LAPP W. S.
Published: June 2006

In previous studies we demonstrated that an induced asialo‐GMI positive (ASGMI+) cell of donor origin that exerts natural killer cell‐like activity (NK activity+) plays a crucial role in the development of graft‐versus‐host (GVH)‐associated tissue damage and severe immunosuppression. This study examined whether the ASGMI+ (NK activity+) GVH effector cells were activated by non‐specific signals or whether these cells were triggered by specific alloantigens and displayed antigenic specificity, C57B1/6 (B6) donor mice were treated with either B6xAF1 (B6AF1) lymphoid cells and anti‐asialo GM 1 antibodies (anti‐ASGM I) to induce and eliminate specifically activated B6–anti‐B6AF1 ASGM1+ (NK activity’(cells or with polyinosinic: ploycytidylic acid (poly I:C), and anti‐ASGMI to eliminate non‐specifically activated ASGM1+ (NK activity +) cells. Donor spleen and lymph node cells depleted of the specific allo‐induced ASGMI+ NK reactive cells showed near normal numbers of L3T4+ and Lyt‐2+ cells and retained T‐ and B‐cell functions as measured by mitogen responses (to PHA. Con A and LPS). mixed lymphocyte responses (MLR) (to B6Ar'i)and the generation of cytotoxic T cells (CTL) (to B6AF1 blasts). Anti‐ASGMI treatment almost completely abrogated NK activity in all donor inocula. GVH reactions were induced by injecting treated donor cells into B6AF1. B6 × C3HejF1 (B6C3HF1) and B6 x SJLF1 (B6SJLF1) hybrids and monitored by splenomegaly, suppression of T‐cell mitogen responses and the development of histopathological lesions in the thymus. liver and pancreas. Cells from donors depleted of non‐specifically (poly 1:C) induced ASGMI+ cells induced severe histological lesions, marked immunosuppression and splenomegaly in all three F| hybrid combinations. When the donor cells were depleted of specifically induced (B6–anti‐B6AF1) ASGMI+ cells and injected into the three F1 combinations they induced splenomegaly in all three but caused severe tissue injury and intense immunosuppression only in B6C3HF1 and B6SJLF1 mice and not in B6AF1, mice. Genetic analysis suggests that the H‐2D (or a closely related) region of the H‐2 complex plays an important role in the activation of the specific GVH effector cells. These results suggest that the cell(s) responsible for splenomegaly are different from the ones that cause severe GVH‐associated tissue damage and immunosuppression although there may be cells and/or lymphokines common to both processes GVH‐associated tissue injury appears to be due to the induction of an effector cell that expresses ASGMI and exerts both NK‐cell activity and specific cytotoxicity but does not appear to be from the T‐cell population that is responsible for specific MLR and CTL activities in vitro.
What Determines an Antigen‐Specific IgE Isotypic Response?—A Hypothesis
Authors: Chen Swey‐Shen Alex
Published: June 2006

Two models to account for an antigen‐specific IgE isotypic response are proposed. Both models assume a first‐tiered IgE production induced by antigen and IL‐4; however, the processed IgE or Ag IgE immune complexes stimulate Tɛ cells differently in the two models. In Model I. we propose that Tɛ cells express conventional T‐cell receptors which recognize IgE isotypic determinants. Model IA proposes that IgE fragments are processed and recognized along with class II MHC molecules, and Tɛ cell preferentially act on antigen‐activated IgE‐committed Tɛ cells via recognition o! processed membrane IgE determinants but not antigens; thus Tɛ cells are in principle capable of modulating non‐antigen‐specific polyclonal IgE responses. Model IB proposes that IgE function as a class‐restriction determinant for nominal antigens analogous to that of class II molecules, and Tɛ cells exert stringent antigen‐specific IgE isotypic responses by recognizing nominal antigens restricted to IgE, Tɛ cells thus exert antigen‐specific and IgE concerted immunoregulation. and do not participate in modulating polyclonal IgE production. Model II proposes a heterotypic interaction of IgE with a cell interaction receptor (or IgE Fc receptor) on T cells, Tɛ cells modulate antigen‐specific IgE isotypic responses via ligation with IgE antigen immune complexes on B‐cell surface; thus. Tɛ cells in principle contribute to polyclonal IgE responses.
Interferons and the Colony‐Stimulating Factors IL‐3 and GM‐CSF Enhance the IFN‐α Response in Human Blood Leucocytes Induced by Herpes Simplex Virus
Authors: CEDERBLAD B. , ALM G. V.
Published: June 2006

Human peripheral blood mononuclear cells (PBMC) were stimulated to produce interferon‐α (IFN‐α by glutaraldehyde‐fixed Herpes simplex virus type I (HSV)‐infected WISH amnion cells in vitro. Different cytokines were included during the stimulation and tested for their ability to enhance the IFN‐α response which occurs in the natural IFN‐α producing (NIP) leucocytes. The total production of IFN‐α and the numbers of IFN‐a producing cells (IPCs) were increased by interleukin‐3 nL‐3) or granulocyte‐macrophage colony‐stimulating factor (GM‐CSF). Their most marked effect was to reduce the time required for induction of the IPC by the HSV‐infected cells, thereby causing both an earlier peak of IPC numbers and secretion of IFN‐α. Addition of IFN‐α2b did not alter the kinetics of the IFN‐α response in the same way as the two CSFs. but instead generally increased the IPC numbers and the production of IFN‐α. The IL‐3 and GM‐CSF, especially in combination with IFN‐α. had the most pronounced enhancing effects on IPC numbers when PBMC were induced at low cell concentrations. The cytokines IL‐1, 1L‐2 IL‐4, IL‐6 or tumour necrosis factor‐α((TNF‐α) had no delectable effects on the IFN‐α response. The results suggest that cytokines such as the CSFs and IFNs may be involved in the regulation of NIP cell functions.
Membrane‐Incorporated Immunoglobulin Receptors Increase the Antigen‐Presenting Ability of B Cells
Authors: GONTIJO C. M. , MÖLLER G.
Published: June 2006

Monoclonal antibodies specific for ovalbumin were conjugated to palmitate and inserted into the membrane of normal spleen B cells. Their presence in the membrane, as well as their ability to bind ovalbumin. was established by immunofluorescence. The so called anti‐ovalbumin‐'decoraled’B cells were tested for their ability to act as antigen‐presenting cells for ovalbumin‐specific I‐Ad‐restricted T‐cell hybridomas. It was found that the antibody‐decorated B cells presented antigen more efficiently than non‐decorated B cells.
Characterization of Monoclonal Anti‐α1‐Microglobulin Antibodies: Binding Strength, Binding Sites, and Inhibition of Lymphocyte Stimulation
Authors: BABIKER‐MOHAMED H. , FORSBERG M. , OLSSON M. L. , WINQUIST O. , NILSON B. H. K. , LÖGDBERG L. , AKERSTRÖM B.
Published: June 2006

Eleven monoclonal antibodies (MoAb) directed against the immunoregulatory plasma glycoprotein α1‐microglobulin were characterized. The MoAb were produced in mice immunized with a mixture of α1‐microglobulin homologues from man. guinea pig, rat and rabbit. Using radioimmunoassay, western blotting, affinity chromatography. and Scatchard analysis, the affinities and binding sites of the MoAb were analysed. All antibodies were more or less cross‐reactive, bul most showed a major specificity for one or two of the α1‐ microglobulin homologues. None of the aniibodies was directed against the carbohydrate moiety of α1‐microglobulin. Six of the MoAb had high affinity for the antigen and four of these were directed towards the same part of the molecule though differing in their species specificity. Five showed lower affinity for the antigen and were mainly directed towards epitopes on other parts of the molecule. Only some of the antibodies could block the proliferation of lymphocytes induced by human α1‐microglobulin. The blocking efficiency of the different antibodies was similar when tested on the stimulation of human or mouse lymphocytes, suggesting that the same part of the α1‐microglobulin molecule is responsible in both species. The magnitude of blocking by the different MoAb was not related to their affinities, emphasizing the importance of where on the α1‐microglobulin molecule, rather than how strongly, they bind. The binding of the strongest blocking antibody was shown lo be directed to a C‐terminal peptide of rat α1‐microglobulin, indicating that this part of the α1‐microglobulin is important for the mitogenic effects. Thus the panel of anti‐α1‐microglobulin MoAb should be a valuable tool for structural and functional studies of α1‐microglobulin.
Dietary Antigen Handling by Mother and Offspring
Authors: TELEMO E. , BATLEY M. , MILLER B. G. , STOKES C. R. , BOURNE F. J.
Published: June 2006

Sows were Ted ovalbumin (OvA) as a novel protein antigen either throughout gestation and lactation (G + L) or during tactation only (L). This resulted in a significant uptake of OvA into blood, colostrum and milk along with a specific IgG response. In piglets from the G + L group. OvA and antibodies to OvA were detected in serum after ingestion of colostrum. In a large proportion of these piglets OvA was still detected at 3 weeks of age. In she L group a significant proportion of the piglets responded to OvA whilst still suckling their mother.

At 3 weeks of age all piglets were weaned onto an egg‐based diet. A similar uptake of OvA was seen in all piglets but there was no response to OvA in the G + L piglets. In piglets from sows fed only during lactation, however, a rapid IgG anti‐OvA response and signs of diarrhoea were seen.

The results suggest that factors of immunological importance are passed over from mother to offspring and it is proposed that immunological experience of dietary antigens by the mother is important for a 'safe’tolerance induction in her offspring.
The IgG FcRII and the PI‐linked IgG FcRIII Trigger Cytoplasmic Calcium Fluxes Independently in Human Granulocytes
Authors: LUND‐JOHANSEN F. , OLWEUS JOHANNA , AARLI Å. , BJERKNES R.
Published: June 2006

The aim of this study was to investigate signal transduction through the two Fc receptors for IgG on human granulocytes. Using How cytometry and the calcium indicator Fluo‐3, we measured changes In leucocyte cytoplasmic calcium concentrations following cross‐linking of cellular Fc receptors with specific antibodies. Two different approaches were used in order to study the two Fc receptors independently of each other. One was lo avoid the presence of IgG Fc fragments. capable of binding to both types of receptors. The other was to use leucocytes from a patient with paroxysmal nocturnal haemoglobinuria (PNH) deficient in granulocyte FcRIII. In contrast to earlier reports, both approaches showed that the two types of IgG Fc receptors on granulocytes are capable of increasing cytoplasmic free calcium concentrations independently of each other. The results suggest that free cytoplasmic calcium ions arc involved in the signal transduction pathway of both types of IgG Fc receptors on human granulocytes.
Phagocytosis of β‐1,3‐D‐Glucan‐Derivatized Microbeads by Mouse Peritoneal Macrophages Involves Three Different Receptors
Authors: KONOPSKI Z. , RASMUSSEN L.‐T. , SELJELID R. , ESKELAND T.
Published: June 2006

Intraperitoneal injection of β‐1,3‐D‐glucan coupled to the surface of monodisperse methacrylate microbeads improves the resistence against bacterial infections in mice, while methacrylate microbeads alone do not. The effect of the glucan‐derivatized microbeads (GDM) is considered lo be mediated through peritoneal macrophages [34, 40].

We show that both GDM and (he underivatized methacrylate microbeads (UDM) treated with normal serum were rapidly bound and phagocytozed by mouse peritoneal macrophages in vitro. We found that both complement and fibronectin opsonized the bead.‐; and were responsible for the uptake.

Treatment of microbeads with serum lacking fibronectin and complement activity still gave BOOK uptake of GDM, but not uptake of UDM. The uptake of GDM was similar to the uptake of untreated GDM and was inhibited by pretreatment of macrophages with soluble β‐1,3‐D‐gluean.

Our conclusion is that GDM and UDM intraperitoneally bind fibronectin and C3 through activation of the alternative pathway of complement. This leads to their phagocytosis by macrophages through fibronectin and complement receptors. GDM are also internalized via β‐glucan receptors. We present the hypothesis that the β‐glucan receptors on peritoneal macrophages account for the protective effect of GDM in intraperitoneal bacterial infections.
Mechanism of Calcium Ionophore and Phorbol Ester‐Induced T‐cell Activation
Authors: HASHIMOTO S. , TAKAHASHI Y. , TOMITA Y. , HAYAMA T. , SAWADA S. , HORIE T. , MCCOMBS C. C. , MICHALSKI J. P.
Published: June 2006

We examined (he role of monocytes in T‐cell activation induced by phorbol myristate acetate (PMA) and calcium ionophore ionomycin. Depletion of monocytes from peripheral blood mononuclear cells (PBMC) was associated with the loss of inlerleukin‐2 (IL‐2) production, IL‐2 receptor (1L‐2 RI expression and proliferation, in response to cither PMA or ionomycin. Addition of monocytes to highly purified T cells resulted in the complete R‐constitution of IL‐2 production. 1L‐2R expression and proliferation by PMA‐stimulated lymphocytes. Exogenous IL‐2, but not interleukin‐l (IL‐l), could reconstitute the T‐cell responsiveness. Addition of monocytes to highly purified T cells stimulated with ionomycin resulted in partial reconstitution of IL‐2 production. II,‐2R expression and proliferation. Similarly, (he addition of exogenous lL‐2 to ionomycin‐stimulated T cells only partially reconstituted the response compared with PBMC, These results suggest that monocyte‐T‐cell interactions contribute to IL‐2 production and IL‐2R expression and are crucial events for PMA‐induced T‐cell proliferation. With ionomycin, monocytes play a role, in part, in inducing IL‐2 production, IL‐2R expression and proliferation. However, IL‐2 is not a sufficient signal to induce T‐cell proliferative response lo ionomycin, suggesting that an IL‐2‐independent mechanism may exist in ionomycin‐induced T‐cell proliferation.
Highly Polymorphic Domains of the Human Serum Amyloid A (SAA) Gene GSAA1
Authors: SACK G.H. , TALBOT C.C.
Published: June 2006

The DNA of the human serum amyloid A (SAA) gene GSAA1 contains several repetitive regions within its introns. We have studied length variations al one such region in the 2nd intron by selective amplification using the polymerase chain reaction (PCR) and defined oligonucleotide primers. The lengths of the repetitive regions frequently differ between individual chromosomes and can be transmitted as Mendelian markers, making them useful for genetic linkage analysis.
Defective Candida Opsonin and Procidin Function in Patients with Recurrent Pyogenic Infection: A Common Immunodeficiency
Authors: PARKIN J. M. , BRIDGES J. , SLOPER K. S. , WARNER J. O. , PINCHING A. J.
Published: June 2006

Polymorphonuclear cell function tests were performed in 340 patients with recurrent unexplained infections. Serum‐dependent defects in phagocytosis and intracellular killing of Candida albicans (Candida opsonin and procidin defects) were specifically associated with susceptibility to recurrent pyogenic infections, Candida opsonin defects occurring in 12.7% and Candida procidin defects in 20.8%, of patients compared with 3.7%., of healthy controls. Saccharomyces cerevisiae opsonin deficiencies were also found frequently in patients with recurrent infections, being present in 18.2% compared with 7.4% controls, although not specifically in those with pyogenic infection.
Analysis of Type II Collagen Reactive T Cells in the Mouse
Authors: ANDERSSON M. , CREMER M. A. , TERATO K. , BURKHARDT H. , HOLMDAHL R.
Published: June 2006

The specificity of the recognition of type II collagen (CII) by T cells in the DBA/1 mouse was analysed using fragments of chick and rat CII obtained by cyanogen bromide (CB) cleavage. Firstly DBA/1 mice were immunized with chick CB fragments 5, 8, 9, 10, 11 and 12. Ten days later the draining lymph node cells were cultured with rat and mouse CII and the proliferative response was determined by incorporation of [3H]thymidine All peptides were capable of triggering T cells recognizing rat CII but only CB9 immunized mice responded well to mouse CII. Secondly, lymph node cells from CBA/1 mice immunized with rat and mouse CII were cultured with the CB fragments including rat CB10 and CB11, and the proliferative response was determined. After immunization with rat CH, the response was strongly dominated by T cells recognizing CB11 with equal responses against chick and rat CB11. After immunization with mouse CH only rat CB10 gave a strong response. It is concluded that several epitopes on the CH molecule can be recognized by T cells in the DBA/1 mouse and that most of these epitopes are shared by rat and chick CH but not mouse CH. These epitopes exhibit strong immunodominance. In mice immunized with intact heterologous CH, the immunodominant response is directed against one or more epitopes on the CB11 fragment present on several heterologous CH but apparently not on mouse CH. In mice immunized with autologous CH the immunodominant response is directed against one or more epitopes on the CBl0 fragment present on rat and mouse CH. They are either absent in chick CH or located in the carboxyterminal end of the CB10 fragment where a cyanogen bromide cleavage site is present in chick CH but not in rat CH. These results suggest that the proposed importance of CBH in collagen‐induced arthritis is due to activation of T cells reactive with heterologous CH only. These cells may be important for the induction of the strong auto‐antibody‐response after immunization with heterologous CH. Structures of importance for direct T cell involvement in the arthritic process and recognized by autoreactive T cells are suggested to be found on CB10.
Differentiation of Macrophage Precursors to Cells with LAK Activity under the Influence of CSF‐1 and High Dose IL‐2
Authors: LI Hao , KNIEP E. , EMMENDÖRFFER A. , LOHMANN‐MATTHES M.‐L.
Published: June 2006

Mouse macrophage precursor cells with natural killer (NK) like activity, derived from in vitro culture of light‐fraction‐bone marrow cells in the presence of colony‐stimulating factor 1 (CSF‐l) and low dose IL‐2, were incubated with high dose(1000 U/ml) IL‐2, After 3 days of incubation, cells had developed from NK like (killing Yac‐1 but not P815) into LAK‐like (killing both YAC‐1 and P815) effector cells. Morphological studies revealed that LAK activity occurred at the time when macrophage precursors with NK like activity containing few cytoplasmic granules had further differentiated into cells with abundant azurophilic granules in their cytoplasma. Proliferation of macrophage‐precursor derived NK LAK‐like cells was dependent on the presence of colony‐stimulating factor, generation of cytoplasmic granules was induced by lL‐2 in a dose dependent way. Flow cytometric analysis showed that macrophage precursor‐derived LAK effectors were positive for NK 1.1 but almost negative for F4/80. When the same starting cell population was cultured in the presence of 200 U/ml Interferon gamma (IFN gamma), proliferation was completely stopped and within 3–4 days all cells differentiated into mature macrophages expressing F4/80. In context with our previous data, we describe here a continuum of development from agranular macrophage precursors to granular cells with NK‐like activity and further to cells with LAK activity under the influence of CSF‐I as growth factor and IL‐2 as granule‐ and cytotoxicity‐ inducing factor.