bluuebunny/crossref_metadata_2025 huggingface dataset:
Dimeric Mouse IgA is Transported into Rat Bile Five Times More Rapidly than into Mouse Bile
Authors: KOERTGE T. E. , BUTLER J. E.
Published: June 2006

The kinetics of the biliary dimeric IgA transport in rats is well documented from studies on animals with cannulated bile duets, while in mice such information rests on the analysis of samples periodically removed from the gall bladder and on serum disappearance rates. We have Quantitatively compared the kineties of IgA transport in these two rodents by using affinity‐purified radiolabeled monomerie and dimeric M315 IgA which specifically binds dinitrophenyl (DNP). Bile collection procedures, sample analysis, and administration of the radiolabeled M315 were the same for both species, although in mice the cystic duct was ligated to prevent reflux of any bile or IgA from the gall bladder into the common bile duel. A pronounced selection for transport of dimeric M315 was seen in both species. When dimeric M315 was administered, it was recovered in the dimeric form from the bile of both species, and 60–80% could specifically bind to DNP‐gelatin. When monomeric M315 was administered, very little was transported, and the IgA recovered in bile had a much lower capacity to bind DNP‐gelatin and appeared to be of low molecular weight. Approximately 60% of the intravenously administered dimeric IgA was transported into bile of both species but biliary transport of the heterologous dimeric IgA into rat bile was five times more rapid than the transport of the dimeric M315 into its homologous species, the mouse. Hence, IgA transport in these species is similar in specificity, integrity of the transported IgA and in the percentage cleared, while differing significantly in the rate of transport. This cannot be ascribed to the species source of the IgA.
Alpha1‐Microglobulin Is Mitogenic for Guinea Pig Lymphocytes
Authors: LÖGDBERG L. , ÅKERSTRÖM B. , SHEVACH E. M.
Published: June 2006

Guinea pig alpha1‐microglobulin (alpha1‐m) was found to exert mitogenic effects on guinea pig lymphocytes, i.e. peritoneal exudate lymphocytes (PEL) or T lymphocytes from regional or mesenteric lymph nodes (LNL). On the other hand, spleen T lymphocytes did not respond to alpha1‐m. Stimulation of LNL required the presence of accessory cells. The stimulatory effect was strain‐specific: alpha1‐m clearly induced proliferation of lymphocytes from Strain 2 guinea pigs, whereas Strain 13 lymphocytes responded poorly or not at all. Moreover, alpha1‐m inhibited antigen‐driven proliferation of guinea pig lymphocytes. None of the effects described were species‐specific, e.g. human alpha1‐m exerted similar effects on the guinea pig lymphocytes. These data indicate that alpha1‐m may be involved in the regulation of lymphocyte activation.
Serum and Plasma Fibronectin Binds to Complement Reacted Immune Complexes Primarily via C1q
Authors: BAATRUP G. , SVEHAG S.‐E.
Published: June 2006

The binding of fibronectin to human Clq, C3b, and complement‐reacted immune complexes (1C) was investigated by enzyme‐linked immunosorbent assays. Microplates were coated with BSA followed by incubation with rabbit‐anti‐BSA IgG or F(ab')2 fragments of rabbit anti‐BSA. Incubation of the solid phase with serum a 37°C caused attachment of Clq and C3b. Addition of EDTA to the serum inhibited the binding of C3b, but not Clq, whereas substitution of the anti‐BSA IgG on the solid phase with the F(ab')2 fragments abrogated the Clq, but not the C3b binding Fibronectin binding was observed after incubation of the solid‐phase IC with serum or plasma at conditions where Clq was also bound, whereas only minor amounts of fibronectin were bound to the solid phase IC via C3b. Purified fibronectin showed a dose‐dependent binding to solid‐phase IC pretreated with Clq or fibronectin‐depleted serum, confirming that the binding of fibronectin to IC largely occurred via Clq. Significantly smaller amounts of fibronectin were bound to solid‐phase IC incubated with plasma instead of serum, despite the higher fibronectin concentration in plasma. This difference was found not lo be due to a fibrinogen‐fibronectin interaction in plasma. Binding of fibronectin to preformed fluid phase IC incubated with serum was demonstrated by SDS‐PAOE analysis of PEG‐precipitated IC.
Chemical Composition of Antigen 60 from Mycobacterium bovis BCG
Authors: FABRE I. , Lcar'HOMME O. , BRUNETEAU M. , MICHEL G. , COCITO C.
Published: June 2006

Antigen 60 (A60). the main thermostable immunogen of tuberculin and PPD. has been purified from Mycobacterium bovis BCG cytoplasm, and identified by crossed Immunoelectrophoresis with anti‐BCG polyclonal antiserum. Two A60 fractions, free lipids and lipid‐conjugated compounds, have been recognized. The free lipids represented about 30% (dry weight), and consisted essentially of C16‐C18, fatly acids, and of phosphatidyl‐inositol‐mannosides. Lipocon‐jugates, upon DEAE‐cellulose chromatography and gel filtration, yielded two main fractions of neutral and polar components. Chromatography of delipidated and deproieinizd A60 on Sephadex G‐100yielded: a high molecular weight fraction (Al, 18%, a lipoglucan of ± 106) and a low molecular weight fraction (B, 10%, a lipopeptidoglycan of ± 104) containing mannose, glucose, and small amount of arabinose. The polysaccharide moieties of fractions Al and B were submitted to aeetylalion, methylation, and acid hydrolysis, and ihe structure of the hydrolysed polymer was deduced by combined gas chromalography/mass spectrometry analysis. The results indicated a branched structure involving 1. 4‐, 1, 6‐, and 1, 4, 6‐linked D‐gluco‐ or n‐manno‐pyranosyl residues. Glucan‐ and peptidoglycan‐bound fatty acids were identified as saturated (C16‐C18) and monounsaturated linear acids (C12, C18). Immunodiffusion on agarose gel indicated that delipidalion and proteolysis did not suppress the ability of A60 to yield immunoprrecipitates with anti ‐A60 antiserum. The high polymer fractions obtained by chromatography on DEAF cellulose and Sephadex G‐100 were also reactive. It is concluded that A60 is made of tree lipids and of hpopcptidoglycans of high molecular weights (106‐107) endowed with immunogenic properties.
Human Alveolar Macrophages Synthesize the Functional Alternative Pathway of Complement and Active C5 and C9 in Vitro
Authors: HETLAND G. , JOHNSON E. , AASEBø U.
Published: June 2006

Attachment of protein to agarose beads cultured with macrophages in protein‐free medium containing 3H‐leucine, shows that de novo synthesis of protein with affinity to the beads takes place. We also found that monoclonal antibodies against human C3c, C3g, and a C9‐neo‐antigen as well as polyclonal antibodies against human C5 and C9, bound to agarose beads that had been kept with the macrophage cultures. Demonstration of C3 derivatives on the agarose beads shows that the essential complement factors of the alternative pathway are synthesized and have been activated by the beads. Deposition of C5 and the detection of a neoantigen of C9 on the beads, indicates that the whole terminal complement pathway has been formed and activated. We conclude that human alveolar macrophages form in vitro the functional alternative pathway of complement, C5 and C9, and we have indirect evidence for synthesis of C6, C7, and C8.
Lymphocyte Activation and Induction of Interferon Gamma in Human Leucocyte Cultures by the Mitogen in Mycoplasma arthritidis Supernatant (MAS)
Authors: KIRCHNER H. , BAUER A. , MORITZ T. , HERBST F.
Published: June 2006

This report succinctly summarizes a number of experiments in which human leucocytes were activated by the mitogen in the supernatant of cultured Mycoplasma arthritidis (MAS). Lymphocytes from a large number of donors were found to be reactive when the two parameters of lymphoproliferation and of production of gamma interferon were analysed. Lymphocytes of cord blood origin were also reactive in lymphoproliferation. Their interferon production was low, but this has been established previously for mitogens in general. During the course of the studies, different batches of MAS were tested, including most recently a purified preparation. All of these were active. Removal of accessory cells adherent to nylon wool column abolished MAS reactivity, whereas it has little effect on lymphoproliferation induced by phytohaemagglutinin (PHA). MAS reactivity was also strongly reduced when monoclonal anti‐HLA‐DR antibodies were added to the leucocyte cultures.
Hepatic Production of Biliary IgM in the Rat
Authors: DAHLGREN U. I. H. , AHLSTEDT S. , HANSON L. Å.
Published: June 2006

Drainage of the thoracic duct resulted in a decrease in the IgA level in rat bile, but at the same time there was an increase in both the total IgM level and the specific IgM antibody activity to Escherichia coli 06 in the bile of animals immunized in the Peyer's patches with these bacteria. The increase in total IgM was significantly higher in animals immunized with ihe E coli 06 than in unimmunized rats. The level of total IgG was not altered during the drainage. IgM antibodies to E. coli 04 given intravenously during lymph drainage did not appear in the bile, whereas specific IgM antibodies to E. coli 06 occurring after active immunization increased in the bile of the same animal. The data elucidate two aspects of the hepatic IgM turnover. First IgM could take the place of IgA in cases of IgA deficiency, and second the IgM might originate from intrahepatic production.
A Hypothesis for the Selection of Available Repertoires: T‐Cell Network Early in the Intrathymic Differentiation
Authors: HERA A. DE LA , TORIBIO M. L. , MARCOS M. A. R. , MARQUEZ C. , MARTINEZ‐A C.
Published: June 2006

T‐cell recognition requires direct cell‐cell interactions mediated by major histocompatibility complex (MHC)‐restricted α‐β heterodimeric receptors (Ti) in association with a constant protein complex termed T3 (TcR, Ti‐T3). Interleukin 2 (IL‐2) promotes growth and maturation of T cells upon binding to high affinity receptors (IL2‐R). They are expressed after the recognition of antigen on accessory cells through the TcR [44]. Furthermore, current hypotheses propose that T‐cell interactions are also mediated by a group of T‐cell antigens, particularly T4/L3T4 and T8/Lyt 2 [35], and perhaps Tγ [15]. All their encoding genes are rearranged and/or expressed sequentially during thymocyte differentiation [5, 8, 40, 41, 46, 51, 52]. Thus, developmental analyses of T‐cell function are essential to gain insight into the mechanisms for selection of available repertoires, one of the central problems in immunology.
The Cytotoxic T‐Cell Response to la Antigens: Lack of Correlation between the Level of Specific Cytotoxicity Obtained in Primary Mixed Lymphocyte Reaction and the Frequency of Specific Cytotoxic T Lymphocyte
Authors: BERETTA A. , DEMOYEN P. LANGLADE , LARSSON E.‐L.
Published: June 2006

We have investigated the cytotoxic response to allogeneic I‐E and I‐A antigens in bulk culture and limiting dilution experiments. A high degree of specific killing could be generated from unprimed T cells of I‐E‐negative strains upon stimulation with cells expressing I‐E. In such conditions, cytotoxic T lymphocytes (CTL) were generated in the absence of a specific proliferative response. The frequency determinations by means of limiting dilution experiments showed that I‐E‐specific CTL precursors were much less frequent than the K. and D specific precursors. The results suggest the existence of a population of I‐E‐specific CTL that failed to grow under conditions that allowed the growth of class I‐specific CTL.
Phagocytosis of Agarose Beads by Receptors for C3b (CR1) and iC3b (CR3) on Human Alveolar Macrophages Cultured on Fibronectin in Vitro
Authors: JOHNSON E. , LARSEN T. , HETLAND G.
Published: June 2006

We studied the phagocytosis of agarose beads by human alveolar macrophages in terms of the morphology, the receptors involved, and the cellular substrates (plastic or fibronectin) used. Heads coated with C3b (58%) and iC3b (42%) by treatment with serum, were ingested during 45 min by CR1. and CR3, on the macrophages. This ingestion was inhibited 80‐90% by the presence of polyclonal F(ab)2 anti‐C3 fragments Since the phagocytosis of both C3b‐coated beads was about threefold stronger than for C3b‐coated beads (trypsinized serum‐treated beads), the results indicate that the CR3 is more phagocytic than the CR1. The phagocytosis of initially complement uncoated beads, which are slowly opsonized [4] with macrophage‐produced C3b and iC3b in vitro, was also strongly inhibited (70‐80%) by the presence of anti‐human C3 F(ab)2 fragments. There was an increased phagocytosis (10‐17%) of complement precoated beads by macrophages cultured on the fibronectin substrate versus the plastic substrate. The morphology and rapid phagocytosis of the complement precoated beads was demonstrated by SEM. The general impression was that membranous protrusions stretched towards the beads, which became increasingly enclosed by plasma membrane.
Polyclonal Lymphocyte Responses to Murine Trypanosoma cruzi Infection
Authors: MINOPRIO P. M. , EISEN H. , FORM L. , D'IMPERIO LIMA M. R. , JOSKOWICZ M. , COUTINHO A.
Published: June 2006

Lymphoid activity was studied in spleen and lymph node cells from Trypanosoma cruzi‐infected mice. Blast transformation in each lymphocyte class was assessed by dual parameter analysis for size and surface markers by both FACS and conventional immunofluorescence, while proliferative activity was measured by tritiated thymidine uptake, autoradiography, and analysis of DNA content in single cells. Acute infection results m rapid Mast transformation and proliferative activity of all three lymphocyte classes (Ig+, L3T4+, and Lyt 2+). At 2 weeks of infection most cells in these organs are enlarged and more than half are dividing. By 2 and 6 months after infection (chronic phase of resistant strains), large numbers of activated B lymphocytes and. to a lesser extent, of Lyt 2+T cells are still detected. Similar results were obtained in C57BL/(resistant) and C3H/HeJ (susceptible) mouse strains. The implications of this massive polyclonal lymphocyte response to the parasite for the physiopathology of acute and chronic infection are discussed.
Polyclonal Lymphocyte Responses to Murine Trypanosoma cruzi Infection
Authors: MINOPRIO P. M. , COUTINHO A. , JOSKOWICZ M. , D'IMPERIO LIMA M. R. , EISEN H.
Published: June 2006

Imraperitonal infection of young adult C57BL/6 males with 105 blood or cloned culture forms of Trypanosoma cruzi (CL. strain) induced the appearance in spleen, blood, and lymph nodes of cytotoxic effector cells detectable in a lectin‐dependent 51Cr‐release assay. The effector cells were conventional cytotoxic T lymphocytes (CTL). since they were Thy 1+ and Lyt 2 +. and the lysis of tumour target cells was strictly dependent on the presence of lectin. CTL activity is already detectable in spleen 2 days after infection, reaches a peak at 2 weeks, and returns to normal levels during the chronic phase (1 month onwards). Increased levels of CTL activity were also detected in lymph nodes with similar kinetics, even in animals that were splenectomized prior to infection. In contrast to spleen, significant levels of CTL activity' persisted in lymph nodes in the chronic phases. This functional variable correlates with the appearance of high numbers of large Lyt 2+ lymphocytes in the same organs (50 to 100‐fold higher than in control, uninfected mice). Very similar responses are detected in a T. cruzi sensitive mouse strain (C3H/HeJ). It appears, therefore, that T. cruzi infection results in a large polyclonal activation of Lyt 2+ lymphocytes, some of which differentiate to effector, cytolytic functions.
Degradation of Solubilized [3H] Elastin by Intact Human Monocytes Stimulated by Immune Complexes in Vitro
Authors: JENSEN H. S. , KJÆRSGAARD E.
Published: June 2006

A simple assay is described that permits determination of elastolytic activity released from human in monocytes. The substrate, solubilzed [3H]elastin, is coaled onto cell culture dishes, Blood monocytes are allowed to adhere, and during subsequent serum‐free culture, radioactive degradation products of elastolysis are released into the culture medium. Using this assay we have demonstrated an elastolytic activity by normal human monocytes of 20+2 ng elastin/hour/3X106mononuclear cells. Contaminating granulocytes and lymphocytes do not cause any significant elastolysis. On stimulation with immune complexes in vitro, the monocytes respond with an enzymatic burst that is 3.6 times the basic enzyme activity. This assay could he a useful tool in the evaluation of elastolytic activity of cells from other sources as well as from human monocytes.
Dysfunctional C8β Chain in Patients with C8 Deficiency
Authors: TSCHOPP J. , PENEA F. , SCHIFFERLI J. , SPÄTH P
Published: June 2006

Two sera from unrelated individuals, each lacking C8 activity, were examined by Western Hint analysis, Using antisera raised against whole C8, the two sera are shown to lack the C 8γ chain. indicating a C8γ deficiency, which is frequently observed in cases of dysfunctional C8. In contrast, by means of a specific anti‐C8‐γ anliserum, a C8γ‐like polypeptide chain of apparently identical molecular weight compared to normal C8γ was detected. Digestion of normal and dysfunctional C8γ with Staphylococcus aureusV8 protease revealed distinct differences in the enzymatic digestion pattern. We conclude that the dysfunction in the C8 protein in these two patients resides in the dysfunctional C8γ chain, and that this form of C8 deficiency is distinct from C8 deficiencies previously reported, in which one or both C8 subunits are kicking.
Immunoregulatory Function of T8 and T4 Cells from Synovial Fluid and Blood of Patients with Rheumatoid Arthritis or Other Forms of Chronic Arthritis
Authors: NILSSON E. , STEDINGK L.‐V. VON , BIBERFELD G.
Published: June 2006

The suppressor effect of synovial fluid (SF) T8 cells and blood T8 cells on the pokeweed mitogen (PWM)‐induced T4 cell‐dependent immunoglobulin production of autologous blood B cells was studied in nine patients with chronic rheumatic diseases (six patients with rheumatoid arthritis (RA), one patient with juvenile RA, and two patients with other forms of chronic arthritis). The suppressor effect of SF T8 cells was of the same magnitude as that of equal numbers of blood T8 cells from patients and healthy controls. However, the relative number of T8 cells was higher among SF T cells than among blood T cells in several cases. Good synovial T8 cell suppression was also demonstrated in coculture experiments where SF T4 cells and B cells were used. In PPD (purified protein derivative of tuberculin)‐stimulated cultures the suppressor effect of SF T8 cells as well as of blood T8 cells from patients and controls was lower than it was in PWM‐stimulated cultures. In most patients SF T4 cells showed a much better PWM‐induced helper function than did non‐fractionated SF T cells. Thus the poor PWM induced helper effect of non fractionated synovial T cells was in some cases mainly due to the suppressor effect of T8 cells, whereas in some cases there was also a deficient helper function of synovial T4 cells.
Abrogation of Staphylococcal Enterotoxin A‐Induced Suppressor Cell Activity by the Anti‐Tac Monoclonal Antibody
Authors: CARLSSON R. , HEDLUND G. , SJÖGREN H.‐O.
Published: June 2006

Human mononuclear cells stimulated with staphylococcal enterotoxin A (SEA) for 2–6 days significantly suppress [3H]thymidine incorporation and reduce the levels of interleukin 2 (IL‐2) and interferon (IFN) in culture medium when added to fresh, polyclonally activated mono‐nuclear cells. The inhibitory capacity of the cells correlates well with the expression of IL‐2 receptors. Lymphocytes obtained 3 days after stimulation with SEA, when the IL‐2 receptor expression is high, are more potent inhibitors than cells obtained 2, 6, 11 or 14 days after stimulation, when the IL‐2 receptors are less expressed on lymphoeytes. T4′ T8′ cells were both found to be inhibitory. Irradiation of the cells with 15 Gy before stimulation with SEA reduced but did not climinate their suppressive capcity. The expression of the IL‐2 receptor was lower in the irradiated cells. Irradiation or mitomycin‐C treatment of cells after 3 and 5 days of SEA exposure had no effect on their inhibitory capacities. Pretreatment of the cells with IL‐2 could partially reverse their suppressive effect on recorded IL‐2 levels of stimulated fresh cultures. A complete reversal was obtained with the anti‐Tae monoclonal antibody, which binds to the IL‐2 receptor. The collective data show that the SEA‐included suppression of IL‐2 activity in lymphocyte culture medium is not due to a suppression of the IL‐2 production but rather depends on depletion of IL‐2 due to absorption of IL‐2 from the culture medium.
Age‐Dependent Changes in Isotype Expression and Down‐Regulation of C57BL/6 Mice
Authors: PONNAPPAN U. , CINADER B. , GERBER V. , BLASER K.
Published: June 2006

Age‐related changes in antibody response and tolerance inducibility are polymorphic; in this paper isotype changes in ageing C57BL/6 mice are examined Female C57BL/6 mice of various ages were immunized with either heat‐aggregated RGG (a‐RGG) or phosphorylcholine conjugate of RGG (PC‐RGG); other animals of the same ages were given aggregate free RGG, followed by injections with either aggregated RGG or haptenated RGG. Sera from these four groups were analysed for antibody isotype. The data presented here indicate that age‐related changes in isotype predominance and magnitude are different for different determinants. The capacity to be down‐regulated appeared to undergo different age‐related changes with different isotypes: there is split tolerance in isotypes. Age‐dependent changes in T‐ and B‐cell tolerance could be deduced by comparing responses of animals to hapten and to carrier determinants. In 5‐week‐old animals tolerance to hapten was more profound than tolerance to carrier. It was concluded that T‐cell regulation dominated the response at this age. With advancing age, i.e. by 95 weeks, tolerance is observed in response to hapten but not in response to carrier determinants. We concluded that suppressor cells were induced by aggregate free RGG and affected the response of ‘naive’ but not of ‘experienced’ B cells.
The Protective Effect of β1–3D‐Glucan‐Derivatized Plastic Beads against Escherichia coli Infection in Mice
Authors: SELJELID R. , RASMUSSEN L.‐T. , LARM O. , HOFFMAN J.
Published: June 2006

Pretreatment with β‐1, 3‐D‐glucan‐derivated plastic beads conferred strong protection against Escherichia coli infection in mice. The protective effect showed a dose‐response relationship to the amount of beads injected and was dependent on the time point of the injection relative to the infection with E. coli. A similar protection could be obtained in nude mice. Experiments with radioactively labelled bacteria as well as beads indicated a systemic effect of the beads. macrophages extracted from animals treated with glucan plastic beads appeared highly stimulated. This was also true of cells that did not contain beads and presumably therefore not glucan, which seems to indicate a soluble stimulatory factor.
The Six Equilibrium Binding Parameters of Two Antibodies in Competitive Radioimmunoassays
Authors: BOJESEN E.
Published: June 2006

A procedure is described for measuring the six parameters that theoretically determine the equilibrium bindings of two ligands in two‐antibody antisera used in competitive radiom‐munoassays (RIA). Four intensity parameters and two quantities are identified. They are the dissociation constants of the complexes formed by the bindings of the tracer and the specific ligand to two antibodies and the two corresponding binding capacities of a certain volume of the antiserum. Four series of assays, runs, are required to give enough information. Two very different amounts of serum are used, either with various doses of the tracer only or with one tracer dose and suitable dose ranges of the specific ligand. The corresponding equations and their use are presented. The amount of information is sufficient to offer several controls of consistency of the parameters with the two‐antibody model. The RIA antisera of two pro‐staglandins, PGF 2α and PGE 2 with the heavily tritiated analogues as tracers were found to be very different with regard to the ratios of the binding capacities and the affinities of the two antibodies. The intensity parameters of the two sera were stable over the period of the present study, almost a year. To interpret assay results of a run of unknowns it is necessary to calibrate precisely the amount of antiserum used as well as the residual non‐specifically bound tracer fraction and the tracer dose. The random analytical error of an assay is then also predictable when the amounts of tracer or antiserum vary between assays of the same run.
Stages in LAV/HTLV‐III Lymphadenitis
Authors: GERSTOFT J. , PALLESEN G. , MATHIESEN L. , DICKMEISS E. , LINDHARDT B. ØRSKOV , HOFMANN B. , NIELSEN C. M. , PETERSEN C. S. , KROON S.
Published: June 2006

The aim of the present study was to investigate the relation between the histopathological findings in LAV/HTLV‐III lymphadenitis and immunological, clinical, and serological variables. The study group included 38 consecutive homosexual men with persistent generalized lymphadenopathy (PGL) in whom lymph node biopsy was performed. The histopathological lymph node changes were grouped into three stages. Opportunistic infections at the time of biopsy and their development during follow‐up were significantly associated with stage III histology (follicular depletion). Analysis of blood from 10 patients with stage III histology revealed significantly (P0.01) decreased proliferative responses of lymphocytes to mitogens and reduced absolute number of CD5+ and CD4+ lymphocytes compared with 17 patients with stage I histology (follicular hyperplasia), whereas patients with stage II histology (follicular involution) had intermediate values. The absolute number of CD8+ lymphocytes was increased in all three stages, as was IgG, while increase in IgM and IgA was restricted to stage III. No difference was observed between the different histopathological stages with respect to the specificity of the anti‐LAV/HTLV‐III antibody as measured by immunoblotting. In conclusion, the defects of lymphocytes from teh blood of LAV/HTLV‐III infected persons reflect alterations in secondary lymphoid tissue. Further, there is a close correlation between these alterations and the clinical status of the patients.
Immunoregulatory Lymphokines in Rheumatoid Joints
Authors: EGELAND T. , LUND H.
Published: June 2006

Since there have been conflicting findings about the presence of interleukin 2 (IL‐2) in synovial fluid (SF) of patients with rheumatoid arthritis (RA), we decided to study this subject ourselves. In a sensitive assay utilizing CTLL‐2 as indicator cells, no IL‐2 like activity was found in SF from any of the 15 RA patients studied. Treatment of SF with hyaluronidase, dialysation, heating to 56°C for 1 h, preincubation of CTLL‐2 with SF prior to washing and incubation all resulted in no detection of any IL‐2. SF itself did not inibit the CTLL‐2 assay. Failure to detect IL‐2 may to some extent be explained by in vivo absorption of any IL‐2 by IL‐2 receptor‐positive T cells in SF. The failure is discussed along with new evidence from various reports suggesting that impaired IL‐2 production by activated T cells may be a feature of several autoimmune diseases.
In Vitro Induction and Characterization of Mast Cells from Murine Peyer's Patches
Authors: KAWANISHI H. , IHLE J. M.
Published: June 2006

The present study was carried out in vitro to determine whether murine Peyer's patches (PP) can be a source of mucosal mast cells, using a well‐defined mast cell growth factor, interleukin 3 (IL‐3). The non‐T, non‐B, non‐adherent (null) cell population of PP contained by the precursor cells for mast cells. The mast cells induced possessed toluide blue and alcian blue (pH 3)‐positive granules, which were heterogeneous and contained histamine. The IL‐3‐induced mast cells were IL‐3‐dependent, did not express la, T‐cell, B‐cell or macrophage markers, and released histamine in response to Ca ionophore, but not in response to compound 48/80. These cells bore IgE‐specific surface receptors and intracellular 20 alpha hydroxysteroid dehydrogenase. The latter enzyme activity was induced by IL‐3 in null cells. All these features of the mast cells support the view that these cells belong to a lineage of mucosal mast cells (MMC) and are no different from those of other lymphoid tissues, such as the bone marrow and spleen. In the analysis of MMC precursor frequency, PP had a similar frequency to that of other gut‐associated lymphoid tissues (mesenteric lymph nodes and lamina propria), but was much lower when compared to those of the bone marrow and spleen. Furthermore, like IL‐2, PP are capable of producing IL‐3 in vitro by activating their helper marker‐bearing T cells. Thus, under certain conditions, such as the initial participation by PP in MMC‐associated intestinal immune responses, PP are likely to provide MMC in their microenvironment, like mucosal T and B cells.
Mobile Blood‐Derived Components in Rat Renal Allograft Contain Significant Immunogenic Potential
Authors: HALTTUNEN J. , HÄYRY P.
Published: June 2006

Using a modified ‘primed rejecation assay’ (PRA), we have investigated the immunogenic potential of different rat renal allograft coponents. Temporary transplantation of an extensively perfused DA kidney to a WF recipient can sensitize the WF recipient in less than 10 min; consequentl, the survival of a subsequent DA heart allograft is significantly reduced. Irradiation of the renal allograft donor siginificantly reduces the immunogenic potential of the first allograft, but does not deplete it entirely. When the first allograft was made chimeric with regard to the (capillary) blood compartment, it was found that in vivo perfusion of an irradiated DA kidney with WF blood did not alter the immunogenic potential in WF recipients, whereas perfusion of irradiated DA kidneys with DA blood incresed the immunogenicity. On the other hand, short in vivo perfusion of irradiated WF kidneys with DA blood make the first WF kidney allograft strongly immunogenic in the WF recipient, whereas perfusion of the kidney with WF blood has no effect. The rapid rate of sensitization observed, coupled with the induction and alteration of the allograft immunogenic potential by manipulation of blood‐dervied capillary compartment, suggets that mobile blood‐derived components, which are outside the reach of extensive perfusion, contain siginificant immunogenic potential.
Differential Expression of DRw52‐Like Determinants Detected by Monoclonal Antibodies
Authors: KONING F. , GIPHART M. , TERMIJTELEN A. , BRUNING H.
Published: June 2006

The reactivity of three monoclonal antibodies (MoAb) directed against DRw52‐like determinants was studied in relation to the reactivity of an anti‐DR MoAb using fluorescence‐activated cell sorter (FACS) analysis. The MCS‐7 MoAb reacted with all DRw52+cells and in addition with DR2+, DR4+, and a DR7+ cell. Both the I‐LR2 and the 7.3.19.1 MoAb reacted with DR3+, DR5+, and DRw6+ cells only. However, whereas the 1‐LR2 MoAb reacted strongly with all those cells compared with the anti‐DR MoAb, the 7.3.19.1 MoAb reacted strongly with DR3+ cells. The implications of this for the location of DRw52‐like determinants on DRβ chains is discussed.
Occurrence of Specific Phytohaemagglutinin‐ Reactive Immunoglobulins in the Sera of Certain Individuals
Authors: NUNEZ‐ROLDAN A. , FELD J. , DE CORDOBA S. RODRIGUEZ , WINCHESTER R. J.
Published: June 2006

While investigating sera for possible antibodies to TL‐like determinants, multiparous sera were selected that had specific cytotoxic rectivity against phytohaemagglutinin (PHA)‐activated lymphocytes of certain individuals and were not correlated to any HLA specificities. However, in an indirect immunofluorescence assay with flow eytometry, these same sera had strong specific monomorphic rectivity to PHA‐activated lymphocytes from all the indiviuals in the panel. Unstimulated cells remained negative. In contrast, other human sera lacked rectivity with PHA‐stimulated lymphocytesl. The addition of PHA to fresh lympocytes followed by incubation at 4°C for 30 min resulted in the same pattern of reactive and non‐reactive sera. When incubated with PHA, different cell types, including 0 erythrocytes and a murine lymphoid cell line, reacted similarly with the sera. Using plates coated directly with PHA‐E, L, and P in a cellfree ELISA system. PHA‐specific sera reacted specifically with PHA‐coated wells. The anti‐PHA activity was removed by absorption with 0 erythrocytes coated with PHA without affecting the titre of the anti‐HLA antibody in the same serum. These findings suggested that IgG molecules of certain sera react directly with residual PHA bound to the cell sufface and not necessarily with molecules of cellular origiin induced by exposure to PHA, complicating the search for antibodies specific for T‐cell activation antigens.
Loss of Antigen Recognition and Impaired Cytolytic Function in Most Hybrids between Cytotoxic T Cells and BW5147
Authors: WOODLAND D. , JANKOVIC D. L. , WELTZIEN H. U. , McMASTER G. , EPPLEN J. T. , EICHMANN K.
Published: June 2006

In this paper we demonstrate that the generation of cytotoxic T‐cell hybridomas (CTH) that retain classically H‐2 restricted and antigen‐specific lytic activity is not generally feasible by fusion of cytotoxic T cells (CTL) with the AKR thymoma BW5147. However, CTH that express a nonspecific lytic activity are redily generated, as revealed by lectin‐dependent assays. We analyse several such hybridomas for the nature of their lytic activity and for the expression of various function‐associated T‐cell molecules. We show that the lytic activity retained by CTH is atypical in that only P815 mastocytoma cells, and no other tumour targets, are lysed in the presence of phytohaemagglutinin (PHA). However, in other respects this lytic activity resembles that of normal CTL: it requires cellular contact, results in a reduction of the viability of the target cells, and is sensitive to both EDTA and low temperature. Lytic CTH express low amounts of T‐cell antigen receptor (TCR), whereas non‐lytic CTH have no detectable TCR on their surface. Most CTH, lytic or non‐lytic, fail to express Lyt 2. We conclude that the loss of antigen specificity in CTH is due to reduced expression of T‐cell antigen receptor as well as of other functionally relevant molecules. We further conclude from this work that a search for a better fusion partner may be indicated to facilitate the reliable production of cytotoxic T‐cell hybridomas.
Induction of Various HLA Class II Molecules in a Human Colonic Adenocarcinoma Cell Line
Authors: SOLLID L. M. , GAUDERNACK G. , MARKUSSEN G. , KVALE D. , BRANDTZAEG P. , THORSBY E.
Published: June 2006

The colonic carcinoma cell line HT‐29 had no constitutive expression of HLA class II molecules. Gamma interferon (IFN‐γ) induced expression of HLA class II molecules in a dosedependent manner with 100 U/ml as an optimal dose. The expression of HLA‐DR, HLA‐DP, and HLA‐DQ molecules seemed to follow different kinetics. While DR and DP molecules were maximally induced after 2 days, DQ molecules appeared later with maximum percentage positive cells after 8 days. Treatment with a prostaglandin synthesis inhibitor (indomethacin) neither induced class II expression nor altered the dose‐response curve for IFN‐γ; this indicated that possible endogenous production of prostaglandins iin this cell line did not interfere with its class II expression. The lectins phytohaemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM), and wheat germ agglutinin (WGA) did not induce class II expression.
Generation of the Antibody Repertoire in Individuals with Multiple Immunoglobulin Heavy Chain Constant Region Gene Deletions
Authors: HAMMARSTRÖM L. , CARBONARA A. O. , De MARCHI M. , LEFRANC G. , LEFRANC M. ‐P. , SMITH C. I. E.
Published: June 2006

Antibodies against protein antigens are largely restricted to the IgG1 cubclass in man, whereas anti‐carbohydrate antibodies, at least iin adult, are almost exclusively confined to the IgG2 subclass. In IgG2‐deficient donors where the Cγ2 gene is retained in the genome, antibodies against most polysaccharide antigens are absent. We therefore undertook a study of the antibody repretoire iin 11 adult donors withi immunoglobulin heavy chain constant region gene deletions, homozygous or heterozygous defects, encompassing the Cγ2 gene. In all cases, antibodies against polysccharide antigens were present and restricted to the remaining subclasses (IgG1 and/or IgG3). These results suggest and an unrestricted use of the available VH gene repertoire in donors lacking the Cγ2 gene, and imply that the limited antibody repertorie found in IgG2‐deficient individuals with a retained Cγ2 gene may be a consequence of an altered regulatory mechanism or a structural VH gene defect. However, furthermore, the delection of multiple Cγ2 heavy chain constant region genes did not appear to decrease the IgG switch probability as much, since total serum levels of IgG appear to be normal.
Induction of Cytotoxic Capacity by Recombinant Gamma Interferon in Human Myelomonocytic Leukaemia Cell Lines
Authors: KUOSMANEN M. , SARELIN H. , ANDERSSON L. C.
Published: June 2006

We have investigated the ability of human recombinant gamma interferon (IFN‐γ) to induce functional differentiation in three human myclomonocytic cell lines U937, RC2A, and ML‐2. Treatment with IFN‐γ induced natural killer (NK) cell like cytotoxicity against K‐562 cells in ML‐2 and RC2A but not in U937. U937 and RC2A displayed a spontaneous antibodydependent cell‐mediated cytotoxicity (ADCC), which against nucleated target cells was significantly increased in U937 but not in RC2A after treatment with IFN‐γ. ML‐2 did not display ADCC against nucleated targets either before or after IFN‐γ treatment, but lysed efficiently antibody‐coated erythrocytes. All three cell lines displayed enhanced ADCC against erythrocytes after IFN‐γ treatment. Spontaneous phagocytosis of erythrocytes was seen in U937, and this was enhanced by IFN‐γ treatment, while ML‐2 and RC2A were phagocytically inactive before and after treatment with IFN‐γ. The differentiated functions induced by IFN‐γ treatment in this panel of phenotypically closely related cell lines offers an interesting model for further studies on the IFN‐γ regulated gene expression. Moreover, the increased cytolytic capacity after exposure to IFN‐γ might have implications on the use of IFN‐y for treatment of myelomonocytic malignancies. In such cases, IFN‐γ might even increase the aggressiveness of the tumour.
The Role of I‐A/E Molecules in B‐Lymphoeyte Activation
Authors: FORSGREN S. , MARTINEZ‐A C. , COUTINHO A.
Published: June 2006

We have previously demonstrated that monoclunal anti‐I‐A/E antibodies inhibit B‐cell responses lolipupolysaccharide (LPS). In the present report, the inhibitory effects were shown to be carried out directly on B cells. and to be totally independent of the LPS concentration used, thereby showing that antibodies do not mediate their effect through blocking of accessory cells or steric hindrance of LPS‐receptors. Of the three different phases in B‐cell activation/ induction, proliferation, and maturation, induction was shown to be the most sensitive to inhibition by anti‐I‐A/E antibodies. Thus, kinetic studies showed that anti‐I‐A/E antibodies are only inhibitory for the first 16 h of LPS activation, after which B cells can no longer be inhibited by these antibodies. Class II MHC molecules appear, therefore, to be part of a membrane molecular complex which regulates delivery of activation signals to resting B ceils. Since it was also shown that this time period corresponds approximately to the time required for B cells to express functional reaciivity to growth factors, we suggest that anti‐I‐A/E antibodies act on resting B lymphocytes to inhibit mitogen‐dependent induction of growth receptor expression.
Genomic HLA‐DQβ Polymorphism Associated with Insulin‐Dependent Diabetes Mellitus
Authors: BRUSERUD Ø. , PAULSEN G. , MARKUSSEN G. , LUNDIN K. , THORESEN A. B. , THORSBY E.
Published: June 2006

Twelve insulin‐dependent diabetes mellitus (IDDM) patients and healthy controls, who all carried the serologically defined DR3 and DR4 antigens, were compared with respect to other HLA polymorphisms. No significant differences between patients and controls were found by typing for HLA‐Dw determinants by homozygous cell typing, nor by studies of their genomic DRβ polymorphism using different restriction enzymes. In contrast, certain DR4‐associated genomic DQβ fragments had a significantly different distribution among the IDDM patients than among the controls. Furthermore, when the distribution of all DQβ‐specific fragments which demonstrated polymorphism in our material was taken into account, nine of the 12 DR3, 4 IDDM patients demonstrated a similar DQβ polymorphism compared with only two out of the 12 DR3, 4 controls (P=0.006; corrected P=0.037). Cells from patients and controls who demonstrated this IDDM‐associated DQβ polymorphism stimulated each other significantly Aless in reciprocal MLC tests, compared with the responses seen when their cells were confronted with cells from the DR3, 4 individuals with other genomic DQβ polymorphisms.
Macrophage Stimulation with Some Structurally Related Polysaccharides
Authors: ARTURSSON P. , EDMAN P. , ERICSSON J. L.‐E.
Published: June 2006

The macrophage‐stimulating properties of some structurally related polysaccharides were studied in vitro. When the polysaccharides were presented to the macrophages in a sterically fixed form, i.e. as microparticles, they induced the release of interleukin 1 (IL‐1) from the macrophages. Microparticulate 1.3‐β‐glucan (curdlan) induced nonspecific macrophage mediated tumour cell killing while l.4‐α‐glucan (starch), 1.6‐α‐glucan (dextran), and 1.6‐α‐mannan were without effect. The corresponding soluble polysaccharides did not stimulate the macrophages. Kinetic studies showed that although IL‐1 was released immediately after stimulation, the macrophages needed a time lag of several days to develop tumour cytotoxicity. The development of cytotoxicity paralleled binding of tumour cells to the macrophages. Resident and inflammatory peritoneal macrophages showed differences in their responses to the polysaccharides. Stationary, resident peritoneal macrophages stimulated by macroparticles secreted high levels of IL‐1 but expressed a low cytotoxic activity, while newly recruited inflammatory macrophages released lower levels of IL‐1 but readily killed the tumour cells. The influence of cyclo‐oxygenase products on the IL‐1 release and macrophage cytotoxicity was also investigated. When cyclo‐oxygenase was blocked with indomethacin, a significantly higher release of IL‐1, and then an increased cytotoxicity, were obtained with 1.3‐β‐glucan stimulated macrophages. The results suggest that microparticulate polysaccharides may be useful for studies on the induction of macrophage differentiation and also for studies on nonspecific cellular immune responses in vitro and in vivo.
Cellular Production of Antibodies Related to the Acetylcholine Receptor in Myasthenia Gravis: Correlation with Clinical Stage
Authors: LEFVERT A. K. , HOLM G. , SUNDÉN H. , PIRSKANEN R.
Published: June 2006

Spontaneous and pokeweed mitogen‐induced production of specific autoantibodies were studied in cultures of peripheral blood mononuclear cells from patients with different clinical stages of myasThenia gravis. Receptor antibody‐related idiotypes and anti‐idiotypic antibodies were defined by binding to mouse monoclonal anti‐idiotypic and anti‐receptor antibodies, respectively. Patients with severe disease had a more complete spectrum of idiotypes in serum, and cells from such patients spontaneously produced more antibody species and higher concentration of both idiotypes and anti‐idiotypes than patients with mild disease. The frequencies of antibody specificities in tissue culture supernatants more closely reflected disease activity than those in serum. Tissue culture for the study of different species of autoantibodies has proved to be a useful tool for monitoring the disease and the effects of treatment.
Characterization of Leukotriene B4‐Omega‐Hydroxylase Activity within Human Polymorphonuclear Granulocytes
Authors: BROM J. , KÖNIG W. , STÜNING M. , RAULF M. , KÖLLER M.
Published: June 2006

Human polymorphonuclear granulocytes (PMN) metabolize exogenous [3H]leukotriene B4 (LTB4) into 20‐hydroxy‐ and 20‐carboxy‐[3H]LTB4. The conversion was enhanced at acidic pH values (pH 6.0–7.0). Sonication of purified PMN and subcellular fractionation by differential centrifugation showed that major LTB4‐hydroxylase activity was associated with the microsomal fraction (105,000 g pellet). In contrast to intact cells. LTB4‐hydroxylase activity within the microsomal fraction revealed optimal activity at neutral pH and was inhibited by a wide range of divalent cations. There was a strict requirement for the presence of suitable electron donors such as NADPH, Heterocyclic nitrogenous bases, such as imidazole and pyridine, inhibited the LTB4, conversion induced by intact PMN as well as by their microsomes. These observations combined with the spectrophotometric analysis (carbon monoxide dithionite‐reduced difference spectrum) supported the assumption that LTB4‐hydroxylase resembled a cytochrome P‐450 enzyme. The LTB4‐hydroxylase within human PMN was not identical with the cytochrome P‐450 of rat liver; hepatic microsomes only showed minute conversion of LTB4.
Cytotoxic Human HLA Class II Restricted Purified Protein Derivative‐Reactive T‐Lymphocyte Clones
Authors: HANSEN P. W. , PETERSEN C. MUNCK , POVLSEN J. VESTERGÅRD , KRISTENSEN T.
Published: June 2006

Human T‐lymphocyte clones specific for antigenic components of purified protein derivative (PPD) of tuberculin were generated by limiting dilution using in vilro PPD‐activated peripheral blood mononuclear cells from a single donor. The HLA restriction specificity of eight clones that were cytotoxic against autologous PPD‐pulsed monocyte targets, was examined against a panel of allogeneic PPD pulsed targets, In agreement with our findings with bulk‐expanded PPD‐reactive cytotoxic T lymphocytes, all clones were restricted by HLA class II antigens: seven by HLA‐DR 2 and one by HLA‐DRwlO‐the other HLA‐DR antigen of the donor. All clones were CD3+, CD44, CD8‐. One clone exhibited, in addition to HLA‐DR2 restriction, unrestricted cytotoxic alloreactivity against HLA‐DR1. In monoclonal antibody‐blocking experiments the latter clone was the only one that was blocked. Its lytic ability was abolished by two monoclonal antibodies against monomorphie HLA DR determinants. The antigen specificity of the clones was studied by using autologous monocyte targets pulsed with antigens prepared from a range of different mycobacterial species. All seven HLA‐DR2‐restricted clones reacted with the majority of antigens tested. In contrast, the HLA‐DRwlO‐restricted clone reacted exclusively with an antigen unique to PPD.