bluuebunny/crossref_metadata_2025 huggingface dataset:
Functional Property of la‐Positive T Cells in Peripheral Blood from Patients with Systemic Lupus Erythematosus
Authors: KOIDE J.
Published: June 2006

la‐positive (la+) J T cells in peripheral blood and their functional property were examined in patients with systemic lupus erythematosus (SLC). Binding of specific monoclonal antibodies was assessed by indirect immunofluorescence. Functional study of la+ T cells was carried out in coculture experiments by measuring the IgG secreted into the culture supernatant. We found that the percentage of la+ T cells in peripheral blood from patients with SLE was raised and the rise correlated positively with serum gamma globulin and IgG level. The elevation was further increased after stimulation with DNA in vitro, indicating the presence of DNA‐sensitive T cells. Functionally. la+ T cells acted as helper cells in spontaneous IgG synthesis of SLE B cells, and were enriched in the OKT4 subset. These results indicate that SLE T cells arc activated in vivo and that the la+ T cells may play a crucial role in the immunoregulatory function. Accordingly, demonstration of la antigens on T cells by monoclonal antibody may provide a useful tool for the measurement of immunological activity in patients with SLE.
Epstein‐Barr Virus‐Transformed B Cell Lines Present M. Leprae Antigens to T Cells
Authors: ELFERINK B. G. , OTTENHOFF T. H. M. , de VRIES R. R. P.
Published: June 2006

We have investigated whether or not Epstein‐Barr virus‐transformed lymphoblastoid B cell lines (EBV‐BLCL) are able to present Mycobacterium leprae (M. leprae) to antigen‐reactive T cell lines and clones. Such EBV‐BLCL would provide us with a homogeneous and unlimited source of antigen‐presenting cells. Antigen‐triggered proliferation of T cells has been studied with co‐cultures either with autologous or allogeneic irradiated EBV‐BLCL. Our results show that EBV‐BLCL are able to present M. leprae as efficiently as peripheral blood mononuclear cells, and that they also function in an HLA‐DR‐restricted fashion. Apart from their possible in vivo relevance, these results have important practical implications, in particular for the generation and study of M. leprae‐reactive T‐cell clones.
Humoral Inhibition of Neutrophil Chemotaxis in Crohn's Disease
Authors: D'AMELIO R. , PALLONE F. , MOLI S. Le , PONTESILLI O. , RICCI R. , MONTANO S. , ROSSI P.
Published: June 2006

In patients with Crohn's disease (CD) we investigated the C3 conversion of zymosan‐activated scrum (ZAS) and looked for the occurrence of chemotactic factor inactivation (CFI). We also studied the cell‐directed inhibitory effect (CDI) of the CD patients' plasma and, in the same group, complement activation and complement‐mediated deactivation. The mean value of ZAS C3 conversion in CD was no different from that of healthy controls, but in steroid‐treated patients it was lower than in untreated CD. CFI occurred in 1 of the 23 CD sera tested, and CDI was observed in 6 out of the 22 patients tested. EDTA C3 conversion was present in 12 patients, and complement‐mediated deactivation was associated with high values of EDTA C3 conversion. Our findings indicate that complement dysfunction and inhihitory factors of neutrophil chemotaxis are present in CD. These findings could explain the defective neutrophil migration into skin windows. Whether they are relevant to the pathogenesis of tissue injury or of infectious complications and are specific for CD, however, remains to be established.
Activation of Human T Lymphocytes by Soluble Protein A
Authors: BEAURAIN G. , NIAUDET P.
Published: June 2006

The proliferative response of highly purified human peripheral blood or tonsil lymphocytes in the presence of soluble protein A (SpA) was investigated. SpA was shown to be a potent mitogen for T lymphocytes from peripheral blood or tonsils. Conversely, the non‐T lymphocyte population from peripheral blood responded poorly to SpA, and SpA did not stimulate non‐T lymphocytes from tonsils. The addition of mitomycin‐treated T lymphocytes from peripheral blood enhanced the response of blood non‐T lymphocytes to protein A. This effect was not found when non‐T lymphocytes isolated from tonsils were cultured with mitomycin‐treated T lymphocytes. The proliferate response of unseparated cells or purified lymphocyte populations was enhanced when adherent cells were added to the cultures. It is concluded that the soluble form of protein A activates mainly T lymphocytes and does not induce B lymphocyte proliferation.
Distinct Effects of γ Interferon on Human B‐Lymphocyte Precursor Cell Lines
Authors: BLOMHOFF H. KIIL , DAVIES C. , RUUD E. , FUNDERUD S. , GODAL T.
Published: June 2006

The effect of γ interferon (IFN‐γ) on proliferation and antigenic characteristics of cell lines belonging to the B‐cell progenitor compartment was studied. We observed a selective effect of recombinant IFN‐γ but not IFN‐α on proliferation of the B‐precursor cell lines Reh and KM3. On day 4, after addition of 400 μ/ml IFN‐γ the [3H]thymidine uptake in these cells was reduced to 60% and 45% respectively, while no effect of IFN‐γ was evident on the proliferation of the more mature B‐cell lines Raji. Ramos, B85, and Daudi. On the other hand, both Reh and Ramos showed induction of major histocompatibility complex (MHO class 1 antigen expression in response to 400 μ/ml IFN‐γ. In contrast to 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA), IFN‐γ did not induce increased MHC class II antigen expression on Reh cells. Taken together, our results indicate that IFN‐γ fulfils distinct functions at different levels in the development of B cells.
Generation of Anaphylatoxin Activity Related to C3a, by Treatment of Human Serum with the Nitrogen Nucleophile N2H4 or the Chaotrope KSCN
Authors: VON ZABERN I. , DAMERAU B. , NOLTE R. , GRÜNEFELD E. , VOGT W.
Published: June 2006

The present study is concerned with the proteolytic processing of complement component C3 in normal human serum treated with N2M4 or KSCN in the presence of EDTA. Upon incubation with these agents. C3 is first converted to the thiolester‐cleaved form (C3i) and thereafter fragmented by factor I. In consecutive, relatively slow steps, spasmogenic and platelet‐aggregating activity is released. The active principle shows characteristics of C3a. First, the pretreated sera deactivate guinea pig ileum and platelets towards the action of C3ahog, but not C5a‐desArghog. Second, the activity is only stable under conditions causing inhibition of serum carboxypeptidase N. such as in the presence of EDTA or of MERGETPA (DL‐2‐mercaptomethyl‐3‐guanidinoethylthiopropanoic acid). Third, the molecular weight deter mined by gel filtration is in agreement with that of C3a. The release of C3a activity requires conversion of C3 to C3i, as well as the complement‐independent generation of proteolytic activity in the pretreated sera. The enzyme releasing C3a activity is a serine esterase probably identical with Hageman factor, kallikrein, or another protease related to the contact system.
Phorbol Ester Induction of Ia and a Shared B‐Cell/T‐Cell Phenotype in Normal but Not Autoimmune (lpr) Mice
Authors: DAUPHINÉE M. J. , TALAL N.
Published: June 2006

Tumour‐promoting phorbol diesters are mitogenic for lymphocytes und induce differentiation of B and T cell lines as well as promyelocytic leukaemia cells. This paper demonstrates that l2‐O‐tetradecanoyl‐13‐acetate (PMA), when cocultured with normal murine bone marrow‐cells (BMC), significantly augments an antigenic cell‐surface determinant called 14D10. This antigen is present constitutively in the majority of bone‐marrow lymphocytes of autoimmune lpr mice. PMA has little enhancing effect when cocultured with lpr BMC. In addition. Ia antigenic determinants are increased by PMA in normal but not lpr BMC. Retinoic acid (RA) and PMA act synergistically both to increase 14D10 and to enhance the stimulatory ability of larger lymphocytes as measured by proliferation in an autologous mixed lyniphocyte reaction (AMLR). We suggest that lpr mice have persistent expression of gene products like 14D10 that are usually repressed in normal adult mice. These gene products can be activated in normal mouse bone marrow by PMA which acts through a CV2+ ‐dependent phospholipid‐dependent C protein kinase. The in vivo enhanced expression of 14D10 in lpr mice suggests activation by some mechanism or factors yet to be described.
Loss and Re‐acquisition of Lytic Function by Cloned Cytotoxic T Lymphocytes: Role of Specific Antigen and Interleukin 2
Authors: OCHOA A. C. , WEE S.‐L. , BACH F. H.
Published: June 2006

We have previously reported the isolation of 2 human allospecific cytotoxic T‐lymphocyte (CTL) elones that can undergo a reversible functional change from a highly lytic phase to a nonlytic quiescent phase and again to a lytic phase. The entire process was found to be regulated by recombinant interleukin 2 (rIL‐2). We have now extended these studies in 3 ways. First, we show that specific alloantigens can also function as a signal to reactivate lytic function in the reverted CTL. However, in contrast to CTL reactivated with rIL‐2, the antigen‐reactivated CTL apparently fail to undergo subsequent cell division. Second, we have also found that this reversion phenomenon is not the same for all CTL tested, as 2 other CTL clones were found that did not revert to the non‐lytie phase when cultured for up to 60 h in rIL‐2‐free medium. Third, the expression of Tac (IL‐2 receptor) was also studied throughout the process of reversion and reactivation. rIL‐2 added to the cell that had reverted to a non‐lytic phase induced, an increase in the expression of Tac receptors during the reactivation phase, to levels greater than those expressed on cells continuously cultured in rIL‐2‐supplemented medium.
Activated T Lymphocytes of the Synovial Membrane in Rheumatoid Arthritis and Other Arthropathies
Authors: POULTER L. W. , DUKE O. , PANAYI G. S. , HOBBS S. , RAFTERY M. J. , JANOSSY G.
Published: June 2006

Immunohistological techniques were used to identify activated T lymphocytes within the synovial membrane of patients with rheumatoid arthritis, using the monoclonal antibody (MoAb) RFT2, which identifies a 40‐k dalton molecule preferentially expressed by T blasts or activated cells. Using this reagent together with a monoclonal ‘cocktail’ that stains all T cells, cell counts on consecutive sections of rheumatoid synovium revealed that up to 50% T lymphocytes were RFT2+ (range 9.3‐50.2%, mean 25.4). Subsequent analysis using combination immunofluorescence demonstrated that over 90% of these activated cells were of the T4+ subset. Furthermore all these cells appeared to be Leu8‐. suggesting that the activated population were exclusively ‘true helpers’ and not suppressor inducers. Studies indicated that 50% of the RFT2+ cells were positive with anti‐Tac MoAb. Comparisons with tissues from other arthropathies demonstrated that this relatively high proportion of RFT2+ cells was a feature restricted to rheumatoid arthritis, although biopsies from patients with psoriatic arthritis and ankylosing spondylitis also contained activated cells. Biopsies of Reiter's syndrome, osteo‐arthritis, and pigmented villonodular synovitis contained no activated cells, no were any seen in sections of normal synovium. The presence in rheumatoid synovial membrane of activated T cells which are only of the T4+, Leu8+ subset adds weight to the suggestion that local immunoregulatory dysfunction contributes to the chronic inflammation of rheumatoid arthritis.
Xenohybridization of Epstein‐Barr Virus‐Transformed Cells for the Production of Human Monoclonal Antibodies
Authors: TIEBOUT R. F. , STRICKER E. A. M. , OOSTERHOF F. , van HEEMSTRA D. J. M. , ZEIJLEMAKER W. P.
Published: June 2006

Transformation of human B lymphocytes, obtained from hyperimmune donors with Epstein‐Barr virus, yields polyclonal cell populations in which a minority of cells produce IgG antibodies of predetermined specificity, whereas the majority of cells produce ‘non‐specific’ immunoglobulin (mainly of the IgM class). Such lymphoblastoid cell lines can be easily propagated in high‐densily cultures. Because cloning at l cell per well is not possible, stabilization of lymphoblastoid cell lines by limiting dilution is not feasible and most newly established lines cease to produce specific antibody within a few weeks. Xenohybrids, resulting from fusion of Epstein‐Barr virus‐transformed cells with NS1 mouse plasmacytoma cells, can be cloned at 1 cell per well. Stable xenohybridoma subclones, producing antibody of [he desired specificity, can be isolated after a series of limiting dilutions. In a model system, we have studied the efficiency of xenohybridization of human lymphoblastoid cells. Using this system, we have constructed IgG anti‐tetanus‐toxoid‐ and IgG anti‐HBsAg‐producing cell lines. Next, we investigated whether transformation with Epstein–Barr virus is essential in such a two‐step procedure or whether a polyclonal stimulator, such as pokeweed mitogen, could also be used. It was found that antibody‐producing xenohybrids can be obtained after stimulation with pokeweed mitogen. However, this latter system is subject to more variations and lacks the advantage of pre‐selection of antibody‐producing cells as compared to xenohybridization after transformation.
Characterization of the Immune Response to an Epitope on Mycobacterium leprae Antigen 7 Defined by a Monoclonal Antibody
Authors: REITAN L. J. , CLOSS O. , HARBOE M.
Published: June 2006

A mouse monoclonal antibody (038D‐C6) was shown by crossed immunoelectrophoresis and radioimmunoassay to react with an epitope on the Mycobacterium leprae antigen 7. This epitope was highly crossreactive with BCG/M. tuberculosis and of a non‐arabinogalactan‐arabinomannan nature. A solid‐phase radioimmunoassay (SPRIA) was applied, based on competitive inhibition by human sera of antigen binding by this anti‐M. leprae monoclonal antibody. Inhibitory activity determined by this assay decreased markedlyupon treatment in both lepromatous and tuberculoid leprosy patients. A correlation was found between the bacterial load of the patient and the inhibitory activity measured in the SPRIA assay. Serum‐inhibitory activity could therefore perhaps be used as a follow‐up test for patients on treatment or as a screening method to detect infective cases. A dot enzyme‐linked immunosorbent assay based, like the SPRIA assay, on competitive inhibition by human sera, was explored as an inexpensive and technically simple alternative also applicable under field conditions.
The Chemokinetic Inhibitory Factor Derived from Chronic Lymphocytic Leukaemia Cells
Authors: SIEGBAHN A. , NILSSON K. , VENGE P.
Published: June 2006

We have recently described a heat‐labile and cell‐directed neutrophil migration inhibitory activity that is present in serum from patients with chronic lymphocytic leukaemia (CLL). The inhibitory activity is produced and secreted by CLL cells in vitro. In the present study the inhibitory activity was partially purified from short‐term cultures of monoclonal leukaemic B‐CLL cells. on gel filtration the calculated molecular weight was apparently 30,000. By anion exchange chromatography, the inhibitory factor was recovered in the fractions that eluted with 0.3 mol/l NaCl. The active material applied to preparative agarose gel electrophoresis migrated towards the anode. The inhibitory factor was totally destroyed by trypsin. In addition it bound to concanavalin A‐Sepharose. These properties indicate that the inhibitory factor is a glycoprotein. Antibodies against the isolated inhibitory factor were raised in rabbits. CLL serum was separated by means of gel filtration and the inhibitory activity was recovered in the pool with a molecular weight of approximately 30,000. The activity was completely neutralized by the antibodies raised against the CLL‐cell‐derived inhibitor, indicating the similarity between this and the serum‐derived inhibitor. We have shown the existence of a new lymphokine, derived from B‐CLL cells, in serum from patients with CLL.
Concanavalin A Induction of Suppressor Activity in the T‐Helper Subset Defined by Monoclonal Antibodies
Authors: DAVIDSEN B. , KRISTENSEN E.
Published: June 2006

Human monocyte‐depleted peripheral blood mononuclear cells were separated in T4+ and T8+ populations by a panning technique. Petri dishes coated with goat anti‐mouse antibodies were plated by peripheral blood mononuclear cells coated by monoclonal antibodies, either T4 or T8. The cell populations were separated into adherent and non‐adherent populations based on binding to the goat anti‐mouse‐coated plastic dishes. The purity of the adherent populations was 96%. T4+ and T8+ populations were used as effector cells in the concanavalin A‐induced suppressor test. The T4+ population revealed a pronounced suppressor activity similar to that exhibited by the T8+ population. This finding was independent of two different sources of monoclonal antibodies, T4/T8 and OKT4/OKT8. The registered suppressor activity in the monoclonal antibody‐defined helper population could not be explained either by a switch of the membrane phenotype from T4+ to T8+ cells or by an increased interleukin 2 consumption of the concanavalin A‐treated cells.
Trans‐stimulation of T Cells: Characterization of Targets and Involvement in Loss of Alloreactivity
Authors: PETIT C. , JULIUS M. H.
Published: June 2006

The rapid loss of alloreactivity within populations of antigen‐primed, in vitro propagated T cells cannot he explained by the appearance of suppressor cells nor by the dilution effect of the proliferative antigen‐specific T cells alone. The involvement of trans‐stimulation in loss of alloreactivity, i.e. the recruitment of non‐antigen‐specific T cells into proliferation in an antigen‐dependent and specific fashion, was assessed. Susceptibility to trans‐stimulation was found to correlate directly with stale of activation at the outset of assay. Large T cells (low buoyant density) hut not small T cells (high buoyant density) are susceptible lo trans‐stimulation. Moreover, in vitro pre‐activation of small T cells by mitogen confers susceptibility to trans‐stimulation. Analysis of lire alloreactivity in Percoll fractions of antigen‐primed lymph node T cells revealed activity in both large‐ and small‐T‐cell fractions with some enrichment in the latter. The small T cells, refractive to trans‐stimulation, are diluted out of the population within the early weeks of antigen‐mediated in vitro propagation, accounting for a rapid loss of considerable alloreactivity. The loss of all delectable alloreactivity within antigen‐selected populations suggests that the slate of activation conferring sensitivity to trans‐stimulation must be maintained, and that neither the antigen nor the culture conditions employed met this requirement.
Interaction of Complement‐Solubilized Immune Complexes with CR1 Receptors on Human Erythrocytes. The Binding Reaction
Authors: JEPSEN H. H. , SVEHAG S‐E. , JARLBÆK L. , BAATRUP G.
Published: June 2006

The binding of complement (C)‐solubilized 125I bovine serum albumin (BSA) anti‐BSA immune complexes (IC) to CR1 receptors on human red blond cells (RBC‐CR1) was studied. The binding of IC to CRI was strongly dependent on the molar antigen lo antibody ratio, and IC formed in moderate antigen excess showed no binding. IC solubilized, in 50% human serum in the presence of autologous RBC bound rapidly lo RBC‐CRI, with maximal binding within less than 1 min at 37°C. Release of CRI‐hound IC under these conditions occurred slowly, requiring more than.30 min. Only binding of ‘partially’ solubilized, e.g., anti C3c (C4c) and conglutinin‐reactive 1C occurred, whereas fully solubilized complexes (IC‐C3dg. C4d) showed virtually no binding. Solubilization of IC in the presence of Mg‐EGTA or in C2‐deficient serum resulted in a markedly delayed binding of IC ti RBC, indicating the importance of an intact classical pathway in preparing the IC for binding to RBC‐CR1. C‐solobilized IC could be absorbed to solid‐phase conglutinin or antibody to C3c abd C4c, and tgese kugabds were able to inhibit the binding of solubilized IC to RBC. Heparin also exerted a marked, dose‐dependent inhibitory effect on the binding of presolubilized IC to RBC‐CR1, whereas the binding was unaffected by the addition of monosaccharides or by the concentration of Ca2− or Mg2− ions.
The Fate of Neonatally Injected Effector Cells of Allergic Encephalomyelitis
Authors: WILLENBORG D. O. , SJOLLEMA P. , DANTA G.
Published: June 2006

Lymphocytes from rats sensitized with basic protein (BP) plus complete Freund's adjuvant (CFA), which produce allergic encephalomyelitis when transferred to adult recipients, fail to induce disease when transferred to 3‐ to 5‐day‐old neonatal rats. The transferred cells do, however persist in the recipients and can be revealed by actively challenging with BP‐CFA later in adult life. Challenge leads to a significantly earlier onset of disease than is seen in control animals. We report here that the cells are long lived and persist in the recipients for at least 9 months. The cells can be demonstrated in the spleen and lymph nodes of recipient animals and can be activated by homologous and cross‐reacting encephalitogenic antigenic preparations but not by antigen in nonencephalitogenic forms. These neonatal recipients, which carry autoimmune effector cells asymptomatically for prolonged periods, may provide a useful model for advancing our understanding of immunoregulatory events in this experimental demyelinating disease as well as the human demyelinating disease multiple sclerosis.
Monocyte‐regulated Hyporesponsiveness of Human Cord Blood Lymphocytes to OKT3‐monoclonal‐antibody‐induced Mitogenesis
Authors: PAPADOUIANNAKIS N. , JOHNSEN S.‐A. , OLDING L. B.
Published: June 2006

OKT3 monoclonal antibody recognizes surface antigenic structures present on all human mature T lymphocytes and is mitogenic for resting peripheral T cells. Recent reports suggest that these structures are linked to the specific antigen receptor of the T cells and play an important rule in T‐cells activation. We have tested the mitogenic action of OKT3 on resting lymphocytes from human newborns, their mothers, and unrelated adults. We found that the prohlerative response of cord T cells to OKT3 is significantly lower than the response of maternal and adult cells at all doses of the antibody tested (5–1000 ng/ml). This difference was not dependent on culture conditions (source of serum, kinetics induced by the OKT3 antibody, or different proportions of adherent cells in peripheral blood mononuclear leukocytes), and could only to some extent be accounted for by differences in the proportions of OKT3‐binding cells between these populations. Removal of adherent monocytes largely diminished the OKT3‐induccd proliferation of maternal and adult cells, by an average of 70–80%. In contrast, it significantly enhanced the proliferation of cord cells. The proliferative response of cord T lymphocytes to the two polyclonal T‐cell activators phytohaemagglutinin and concanavalin A was similar to or greater than that of mothers and other adults.
HLA Class‐II‐restricted Mycobacterium lepraereactive T‐Cell Clones from Leprosy Patients Established with a Minimal Requirement for Autologous Mononuclear Cells
Authors: HAANEN J. B. A. G. , OTTENHOFF T. H. M. , VOORDOUW A. , ELFERINK B. G. , KLATSER P. R. , SPITS H. , DE VRIES R. R. P.
Published: June 2006

This report describes an effective method for the cloning of Mycobacterium leprae reactive T lymphocytes with Epstein‐Barr‐virus transformed autologous B cells as antigen‐presenting cells‐ The two advantages of this method are that it drastically reduces the number of autologous peripheral blood mononuclear cells I (<107 cells) needed to obtain and propagate these T‐cell clones (TLC). and that it enables us to expand individual TLC to large numbers of cells (<108) Thus she major obstacles for the cloning of T lymphocytes‐especially important with regard to patients‐are bypassed. Thus far. TLC Srom three leprosy patients have been established. These TLC are HLA class H restricted in their M. leprae directed response. A marked enhancement in antigen responsiveness was observed after further expansion of several TLC. some of which turned from nonresponder into responder TLC, Four tested TLC display strikingly different antigen recognition patterns when tested against a number of other mycobacterial antigens: one T.LC so far recognizes only M leprae antigens.
Induction of Concanavalin A Dose‐dependent T‐Cell Growth Factor Production by Insertion of T‐Cell Membrane Components into the AKR Thymic Lymphoma BW 5147
Authors: LINDQVIST C. , SHAPIRO I. M. , WIGZELL H.
Published: June 2006

Sendai virus vesicles were used as vehicles for the insertion of various cell membranes into different ceil lines. The transplantation efficiency was measured by using FITC‐labelled concanavalin A (Con A) or monoclonal antibodies against the T‐cell marker Lyt 2 and the major histocompatibility complex product H‐2Dh in a fluorescence‐activated cell sorter. Results indicate that it is possible to transplant mitogen responsiveness to certain cell types. Con A stimulation of T‐cell membrane transplanted BW 5147 showed that it is possible to induce a mitogen dose‐dependent T‐cell growth factor production. Consequently this method appears to he an attractive model for further study of the properties of the membrane structures involved in mitogen triggering of cells.
Fibronectin Binding to C1q Associated with Antigen‐Antibody Complexes in EDTA‐treated Plasma
Authors: SORVILLO J. , GIGLI I. , PEARLSTEIN E.
Published: June 2006

In this report we have investigated the association of fibronectin with antigen‐antibody‐C1q complexes incubated in fibronectin‐depleted and C1q‐depleted plasma. When BSA‐anti‐BSA immune aggregates arc incubated in plasma depleted of both fibronectin and C1q to which‘125I fibronectin has been reconstituted, little radioactivity is bound to the immune complexes. However, pre‐incubation of immune complexes with purified C1q prior to incubation in the plasma causes an approximately l0‐fold increase in the amount of radioactivity bound. The binding of 125I‐M‐fibronectin to preformed antigen‐antibody‐C1q complexes is specific, since the reaction is inhibited by the addition of unlabelled fibronectin but not by ovalbumin When antigen‐antibody ‐C1q complexes are incubated in CIq‐depleted plasma containing physiogical concentrations of fibronectin. and analysed by immunoblotting. fibronectin antigens are detected on the immune Complexes. Identical results are obtained using imune complexes composed of sheep erythrocyte rabbit anti‐sheep erythrocyte C1q (EAC1q) cells, There is no specific requirement for preformed antigen‐antibody‐C1q complexes. since fibronectin can be detected on antigen ‐antibody complexes after incubation in normal human serum or in C1q‐depleted ethylenediamineteraacetic acid (EDIA) serum reconstituted with purified C1q prior to incubation with the complexes, Finally, we also demonstrate that in the presence of C1q, 125I‐fibronectin will associate with soluble antigen‐antibody complexes.
Dialysable Leukocyte Extracts (Transfer Factor) Augment Nonspecifically Keyhole Limpet Haemocyanin and Horseshoe Crab Haemocyanin Skin Reactivity in Unimmunized Human Recipients
Authors: ASHORN R. G. I. , VANDENBARK A. A. , ACOTT K. M. , KROHN K. J. E.
Published: June 2006

Dialysable leukocyte extracts (DLE) may induce marked changes in the immune expression of human recipients. It is unclear whether the conversion of skin reactivity by DLE is due to a donor‐related specific transfer factor or to an antigen nonspecific augmenting factor which enhances a preexisting low‐level response in DLE recipients. In this study, DLE from immunized and unimmunized human and calf donors or saline was administered to 88 medical students. The recipient population demonstrated minimal background responses to the test antigens keyhole limpet haemocyanin (KLH) and horseshoe crab haemocyanin (HCH). The results indicate that the DLE preparations from both immunized and unimmunized donors significantly stimulated skin reactivity but not in vitro responses to both KLH and HCH in the recipient population. The results suggest that these DLE preparations contain an immunologically nonspecific augmentor, which stimulates a preexisting low‐level response in the unimmunized population to become a clearly observable skin reaction.
Production of Interferon‐β by Murine T‐Cell Lines Induced by 10‐Carboxymethyl‐9‐Acridanone
Authors: STORCH E. , KIRCHNER H. , BREHM G. , HÜLLER K. , MARCUCCI F.
Published: June 2006

Besides the established T‐cell property of producing gamma interferon (IFN‐γ), murine T cells additionally possess the ability to produce IFN‐α and IFN‐β when appropriate inducers such as 10‐carboxymethyl‐9‐acridanone (CMA) or Newcastle disease virus (NDV) are used, Inter‐leukin 2 (IL‐2)‐dependent murine T‐cell lines, but not purified resting splenic T cells, responded to CMA and NDV with production of IFN‐α, β, The IFN production by these T cells was not restricted to a special subset, since T cells expressing the Lyt 1+2− and the Lyt 1−2+ phenotype responded to these inducers with IFN production, After prolonged passaging of the T‐cell lines in IL‐2‐containing medium, the ability to respond to CMA with production of antiviral activity was sustained longer than the ability for concanavalin A‐induced WH‐y production. Whereas the NDV‐induced T‐cell supernates contained both IFN‐α and IFN‐β, the induction with CMA resulted exclusively in the synthesis of IFN‐β by the T‐cell lines.
Host‐Reactive Cytotoxic T Lymphocyte Precursors in Long‐lived Fully Allogeneic Mouse Bone Marrow Chimeras
Authors: HEEG K. , REIMANN J. , HEIT H. , HEIT W. , WAGNER H.
Published: June 2006

Fully H‐2‐incompatible chimeric mice were constructed by grafting lethally (950 rad) irradiated germ‐free (GF) CBA (H2k) mice with anti‐Thy 1 antibody plus complement‐treated allogeneic C57B1/6 (B6) (H2b) bone marrow cells. These chimeric mice were kept for more than 11 months, either under GF conditions or under barrier‐sustained specific‐pathogen‐free (SPF) conditions. Controls included nonirradiated, nontransplanted, sex‐ and age‐matched CBA and B6 mice raised under SPF conditions, and syngeneic chimeric mice of the CBA × CBA type kept under GF and SPF conditions. All chimeric mice were completely repopulated with donor‐type lymphoid cells and showed no clinical or histological evidence of graft‐versus‐host disease. From the fully allogeneic chimeric mice, we enumerated the numbers of splenic cytotoxic T lymphocyte precursors (CTL‐p) that could be clonally expanded under limiting dilution conditions in response to third‐party alloantigens, or nonmodified and trinitrophenyl (TNP)‐modified stimulator cells bearing host or donor H‐2 antigens. The existence of high numbers of alloreactive and host‐ or donor‐type H‐2‐restricted TNP‐specific CTL‐p in the spleens of fully allogeneic chimeras indicated almost normal immunocompetence. The surprising finding, however, was that large numbers of host (CBA)‐reactive splenic CTL‐p were inducible under limiting dilution conditions in healthy long‐lived allogeneic chimeras, although these chimeric mice were devoid of any histological or clinical signs of graft‐versus‐host disease.
Rheumatoid Synovial Dendritic Cells as Stimulators in Allogeneic and Autologous Mixed Leukocyte Reactions‐Comparison with Autologous Monocytes as Stimulator Cells
Authors: WAALEN K. , THOEN J. , FØRRE Ø. , HOVIG T. , TEIGLAND J. , NATVIG J. B.
Published: June 2006

Dendritic cells were isolated from peripheral blood, synovial fluid, and synovial tissue of patients with rheumatoid arthritis and from peripheral blood of healthy blood donors on the basis of semiadherence to plastic surfaces, The cells were compared with autologous peripheral blood monocytes with respect to their stimulating capacities in allogeneic and autologous mixed leukocyte reactions (MLR), Dendritic cells from the various compartments stimulated allogeneic T cells 6‐14 times more than monocytes did. Dendritic cells also stimulated autologous T cells 10–24 times more than monocytes did. Evidence in favour of the dendritic cell as the major stimulating cell type in MLR was also found in mixed experiments in which various ratios of dendritic cells and monocytes were used as stimulator cells. Furthermore, the activating structures on the dendritic cells seem to be major histocompatibility complex class II antigens, since anti‐HLA‐DR antibodies inhibited the responses. The results, especially from the autologuus MLR, indicate that dendritic cells are important accessory cells for the various immune responses in rheumatoid inflammation.
Proliferative Kinetics, Phenotype and Cytotoxic Function of Thymic and Peripheral T‐Cell Colonies
Authors: RÖPKE C. , CLAËSSON M. H.
Published: June 2006

Cells produced in murine T‐cell colonies grown in methylcellulose have been studied to evaluate their proliferation, phenotypic differentiation, and ability to give rise to cytotoxic cells. Colony‐forming cells from presumably cortical or medullary thymocytes, separated by peanut agglulination, or from lymph nodes, showed rapid proliferation regardless of their origin, although cortical thymic colony cells showed significantly slower proliferation from day 10 today 12 of culture. Essentially all the colony cells maintained the Lyt‐1+ 2+ phenotype of the colony‐forming cells, and no L3T4+ cells were found. The colony cells were characterized as rather mature cells by being strongly H‐2‐positive, and, except for a fewcells, by lacking the TL antigen. Colony cells maintained the Lyt‐l +2+ phenotype after expansion in liquid culture, and both cortically and medullary derived colony cells showed lectin‐dependent cytotoxic activity against P‐815 tumour cells. We conclude that our assay for growing thymus‐derived T‐cell colonies results in the proliferation of a phenotypically and functionally relatively mature cytotoxic T‐lymphocyte progeny.
Origin and Kinetics of IgA, IgG and IgM Milk Antibodies in Primary and Secondary Responses of Rats
Authors: DAHLGREN U. I. H. , AHLSTEDT S. , HANSON L. A.
Published: June 2006

IgA and IgM antibodies were detected in rat milk after immunization wish ferritin in Peyer's patches (Pp) 1 day after parturition but not after intramammary gland or intravenous immunization. The antibody levels decreased from day 9 to day 17 of the nursing period and were undetectable during a second lactation period. Despite the absence of milk IgM antibodies after intramammary gland or intravenous immunization, the serum levels of the IgM antibodies were similar after all three immunization methods. IgA antibodies were not found in serum after any of the immunization methods. IgG antibodies appeared in serum and milk after Pp. intramammary gland, and intravenous immunization. Milk and serum IgG antibodies from all the Pp‐immunized animals decreased from day 9 to day 17 of the lactation period. After intramammary gland immunization, however, the IgG antibody levels increased in all the milk samples, but only in four of seven sera. The milk and serum IgG antibody levels were lower but still detectable during a second lactation period. Re‐injection of ferritin in the Pp during a third lactation period resulted in higher levels of milk IgA, IgG and IgM antibodies than after the first injection, Rats with serum IgG anlihodies against Escherichia colia 08 naturally present in their gut flora had no corresponding milk antibodies of any isotype The results suggest that milk antibodies of all three isotypes stem from local production in the mammary gland and that blood IgG and IgM antibodies originate at least partly from stimulation in Pp.
The Adjuvant Activity of Nonionic Block Polymer Surfactants
Authors: HUNTER R. L. , BENNETT B.
Published: June 2006

We evaluated the molecular and physicochemical properties of surfaces formed by defined layers of block copolymers which were especially effective as adjuvants or in the induction of granulomas. The copolymers which were adjuvants formed hydrophilic surfaces with a large area. They bound protein in a way which left it particularly accessible to antibody and induced the activation of complement. Copolymers which induced granulomas, in contrast, formed hydrophobic crystalline surfaces. They bound less protein and did not activate complement, but were toxic for macrophages. Their surfaces were found to be similar to those of the mycobacterial glycolipid trehalose‐6.6‐dimycolate or quartz, in that they consisted of regular geometric arrays of hydrophilic and hydrophobic adsorptive domains. These studies demonstrated that changes in the size and arrangement of hydrophilic and hydrophobic blocks in copolymers produce a diversity of surface physicochemical properties which correlate with biologic activity.
The Immune Response to Pneumococcal Polysaccharide Type III is Followed by Increased Production of an Antibody Antifactor
Authors: PASANEN V. J.
Published: June 2006

The study shows that serum from pneumococcal polysaccharide type III (SSSIII)‐immunized and nonimmunized mice that has been absorbed until devoid of anti‐SSSIII antibodies inhibits anti‐SSSIII plaque‐forming cells (PFC) but not antisheep red blood PFC in antibody plaque assay. A large fraction of the inhibitions activity can be absorbed by and eluted from anti‐SSSIII‐Sepharose, but not normal mouse serum‐Sepharose. It has low or no affinity for protein A and concanavalin A The titre of the inhibitor increases, with repeated immunizations and inhibits the anti‐SSSIII PFC of rats or mice of different allotypes. On the basis of these characteristics, I classify the inhihitor as an anti‐idiotypic nonimmunoglobulin factor directed against a common determinant of anti‐SSSIII antibodies of mouse and rat, with an immuno‐regulatory function.
Studies of Monoclonal Antibodies Specific for Major Histocompatibility Complex Products of the Rat
Authors: HEMMI S. , FENNER M. , GALL E. , BINZ H. , WIGZELL H.
Published: June 2006

Polyclonal syngeneic, allogeneic. and xenogeneic and monoclonal syngeneic anti‐anii‐idiotypic antibodies have been produced against previously described monoclonal anti‐idiolypic anti‐bodies with specificity for monoclonal RTI alloantigen‐specific antibodies. The anti‐anti‐idiolypes could again be shown to be highly specific for the monoclonal anti‐idiotype used for the induction of the anti‐anti‐idiotypie antibodies and lo carry the same, or a very similar, idiotype as the original monoclonal idiotypic antibody used lo induce the monoclonal anti‐idiotypic. Among the 30syngeneic and allogeneic and the live xenogeneic polyclonal anti‐anti‐idiotypic antisera and the three monoclonal anti‐anti‐idiotypes. only one polyclonal antiserum showed binding capacity to the corresponding RT1‐eneoded antigenic determinants on spleen cells. All the oilier antibodies were idiotypic but not antigen binding.
Bone Marrow Transplantation in the Rat
Authors: RENKONEN R. , HÄYRY P.
Published: June 2006

We have analysed the cytological structure of inflammation in the different parenchymal target organs during acute graft‐versus‐host disease after bone marrow transplantation in the rat. Inflammation (recovery of increased numbers of white cells) was recorded in the liver, skin, and lungs of the allograft recipient compared with the syngeneic graft recipient from day 5 onwards, accompanied by reduced recovery of inflammatory cells from the gut. The major cytological manifestation of the disorder was an increase in the total number of blast cells, large granular lymphocytes (LGL), and lymphocytes in the liver and skin, and of the number of blast cells and LGL in the lung. At the same time, a depletion of LGL and lymphocytes was recorded in the blood, suggesting migration of these cell components to the site of inflammation.
Antigens of Complement Factor C3 Involved in the Interactions with Factors I and H
Authors: NILSSON B. , NILSSON U. R.
Published: June 2006

This study explores the relationship between defined antigens of the C3 molecule and those surface structures that are involved in interaction with factors I and H. Methylamine treatment at pH 11 followed by neutralization converts C3 into a modified state in which the molecule is optimally susceptible to cleavage by factor I in the presence of factor H. The modified C3 is characterized by an antigenic profile with expression of antigens of the C3(N), C3(S). and C3(D) subsets. These antigenic properties closely mirror those of physiologically bound C3b. suggesting that modification of antigenic expression upon denaturation of C3 reflects a regulatory mechanism for I and H function‐ Immunochemical studies of the alkaline‐denatured C3 suggested that factor H internets with surfaces of'C3l that are situated within the C3e fragment and that are defined by C3(SN) antigens, while factor I predominantly interacts with C3(SN) antigens associated with the C3d fragment and with C3(D) antigens hidden in native C3. Therefore the continuing immunochemical study of this and related problems will require polyclonal and monoclonal antibody reagents reactive not only with the C3(S) and C3(N) but also the C3(D) antigenic subsets.
Interleukin 1 Activity Produced by Human Rheumatoid and Normal Dendritic Cells
Authors: WAALEN K. , DUFF G. W. , FØRRE Ø. , DICKENS E. , KVARNES L. , NUKI G.
Published: June 2006

Dendritic cells (DC) from the synovial inflammatory tissue and peripheral blood of patients with rheumatoid arthritis and from the peripheral blood of normal blood donors were compared with the autologous monocytes for their capacity to produce and release interleukin I (IL‐1). Synovial DC often spontaneously released higher amounts of IL‐1 activity than unstimulated and lipopolysaccharide‐stimulated peripheral blood DC and monocytes. The IL‐1 production by both DC and monocytes increased alter stimulation with bacterial lipopolysaccharide‐ In contrast with synovial DC the peripheral blood DC from both patients with rheumatoid arthritis and normal controls released less IL‐1 activity than peripheral blood monocytes did. Inhibition with an antiserum to IL‐1 revealed that IL‐1 production is important for the accessory activity of the peripheral blood DC. Thus human DC from inflammatory sites and peripheral blood produce IL‐1 activity.
Dendritic Cells from Human Rheumatoid Synovial Inflammatory Tissue and Peripheral Blood as Accessory Cells in Mitogen Stimulation of T Lymphocytes.
Authors: WAALEN K. , FØRRE Ø. , PAHLE J. , NATVIG J. B.
Published: June 2006

Dendritic cells (DC) were purified from the peripheral blood (PB) of normal individuals and from the synovial fluid (SF) and synovial tissue (ST) of patients with rheumatoid arthritis. These cells are strongly HLA‐DR positive and lack B cell, T‐cell, and monocyte markers as well as Birbeck granules. The DC were compared with monocytes and non‐T cells from PB for their ability to act as accessory cells for T‐cell responses to concanavalin A (Con A) and phytohaemagglutinin (PHA). DC from PB, SF and ST were much more efficient accessory cells for the mitogenic responses than autologous monocytes from PB. The mean PHA responses in cpm obtained with DC from the various compartments were 4–20 times greater than the responses obtained with monocytes from PB. The Con A responses obtained when the various DC populations were used as accessory cells were 3–13 times greater than those obtained with monocytes from PB. The mitogenic responses seen wish monocytes were very low. The non‐T cells, which comprise a mixture of cells obtained after removal of T cells, also gave low T‐cell responses to PHA and Con A compared with DC as accessory cells.
Expression of HLA‐DQ Antigens on Keratinocytes in Borrelia Spirochete‐Induced Skin Lesions
Authors: TJERNLUND U. , SCHEYNIUS A. , ÅSBRINK E. , HOVMARK A.
Published: June 2006

Skin biopsies were investigated with two different immunohistochemical techniques. thus revealing HLA‐DO antigens on HLA‐DR‐expressing keratinocytes in the late skin manifestations of a Borrelia spirochete infection. In the early skin lesions only HLA‐DR antigens were present on the keratinocytes.’The invariant γ chain of class II transplantation antigens was observed on keratinocytes in 1:5 of the late cases. Upon penicillin treatment detectable HLA‐DR and HLA‐DQ antigens disappeared completely From the keratinocytes. Furthermore, the mononuclear cell infiltrates dominated by anti‐Leu 1 and anti‐Leu 3a‐reactive cells and containing many cells with markers for activation (HLA‐DR. HLA‐DO. transferrin. and interleukin 2 receptors) diminished markedly. The possibility that the expression of different class II transplantation antigens on keratinocytes might reflect separate functional demands of these cells or an altered immunological reactivity in the host, is discussed. the precise functional role of the temporary expression of the class II antigens on non‐lymphoid cells, however, remains an enigma.
Regulation of the Immune Response to Hepatitis B Virus and Human Serum Albumin
Authors: HELLSTRöM U. , SYLVAN S.
Published: June 2006

The circulatory pool of B cells, from donors immune lo hepatitis B (HB) through natural infection, contained sensitized B cells with the capacity to secrete antibodies with specificity for human serum albumin (USA) when stimulated with purified hepatitis B surface antigen (HBsAg) in vitro The immunoglobulin secretion was dependent upon and regulated by T cells and specifically induced, since it was not obtained in cell cultures from HB‐susceptible donors. Culture supernatants with anti‐USA reactivity also contained specific antibodies to HBsAg (anti‐HBs). indicating that the outer coat of HBV normally provokes an immune response to both the viral antigen and a self component. Perturbation in the regulation of the immune response triggered by USA in association with HBV/HBsAg particles may involve a putative risk for development of chronic HBsAg carriership.
Immunocytochemical Localization of a Medullary Thymic Epithelial Glycoprotein
Authors: ZELECHOWSKA M. G. , LECOMTE J. , POTWOROWSKI E. F.
Published: June 2006

The thymic microesnvironment is known to play a key role in T‐cell differentiation, but the exact nature of the interactions between epithelial and lymphoid cells has not been fully elucidated, With a monoclonal antibody to a thymic epithelial glycoprotein, we report the localization of an antigen specific for medullary epithelial cells of the mouse thymus. This antigen is found in the Golgi apparatus of epithelial cells, and on their borders with adjoining lymphocytes. This location is compatible with the previously reported observation that differentiation signals transmitted to thymic lymphocytes by thymic epithelial cells require actual contact between these two cell types.
Dependency of B Cells on the Presence of Adherent Cells, or Factors Derived from Them, for the Production of Autoantibodies in Vitro in the Absence of Cell Division
Authors: DAENKE S. , COX K. O.
Published: June 2006

Peritoneal cells from untreated mice secrete autoantibodies after 3–4 days of in vitro culture, although the cells do not divide. Here, peritoneal cells enriched for B cells to contain 95% surface Ig‐bearing cells, did not secrete autoantibodies after 3 days of in vitro culture unless plastic‐adherent cells derived from the peritoneal cavity were cultured with the B cells. Cell‐free media, taken from peritoneal adherent cells that had been cultured for 3 days in vitro, when added at a final concentration of 50% in fresh culture medium to purified B cells, substituted for the presence of accessory cells. In contrast to cultures of unfractionated peritoneal cells, little increase in precursor frequency was detected when enriched B cells were cultured in the presence of LPS/DXS. However, the addition of adherent cells, supernatants derived from adherent cells, or cytokines produced by a T‐cell hybrid EL4, resulted in an increased precursor frequency when LPS/DXS was added to the culture medium. Three macrophage cell lines. P3788‐D1. J774. and PU‐5‐IR, when added to purified B cells, augmented the autoantibody precursor frequency detected in vitro. This is strong evidence that potentially autoreactive B cells require one or more types of accessory cells in order to differentiate into autoantibody secretors during culture in vitro. Further, the results provide indirect evidence that interleukin 1 may be a crucial molecule in the differentiation of B cells.