bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in Natural Killer Activities in Experimental Secondary Amyloidosis
Authors: KIMURA K. , OGAWA K. , SATO K. , TOKUHARA S. , OTSUKA M. , MATSUNAGA Y. , TSUBURA E.
Published: June 2006

Natural killer (NK) cell aciiviiy in experimental murine amyloidosis was studied. In CBA/J mice, which show a high incidence of amyloidosis. NK activity was significantly decreased after 1 week of casein treatment. In C3H mice, which show a low incidence of amyloidosis, NK activity was not changed by casein treatment. Pretreatment with lipopolysaccharide in vivo enhanced the NK activities in CBA/J and C3H mice. These increases were not observed after casein treatment. The lowered NK activity of cells from CBA/J mice after casein treatment was restored to the normal range by indomethacine in vitro. Depletion of adherent cells from the spleen cells treated with casein had no effect on NK activity. Single‐cell assay showed that casein treatment impaired the killing but not the binding of NK cells to target cells. After casein treatment, the splenic serum amyloid A (SAA) level gradually increased in CBA/J mice but remained low in C3H mice. NK activity was suppressed by the addition of serum obtained from CBA/J mice treated with casein but not by normal control serum. And partially purified AA protein obtained from the spleen of CBA/J mice treated with casein also suppressed NK activity in vitro.
Anti‐Mycobacterium leprae Antibodies in Urine from Lepromatous Patients Examined by Crossed Immunoelectrophoresis and Radioimmunoassay
Authors: OLCÉN P. , HARBOE M. , WARNDORFF T. , BELEHU A.
Published: June 2006

Precipitating anti‐Mycobacterium leprae antibodies were found in concentrated urine samples from 21 out of 42 lepromatous patients. These antibodies were directed against M. leprae antigens 5, 6, and 7. In a radioimmunoassay for anti‐M. leprae antibodies, 90% of these patients had higher antibody levels in their urine than control persons. There was a positive correlation between anti‐M. leprae antibody levels in serum and urine. The advantages of using atraumatically collected samples like urine in epidemiological work are pointed out. The present report shows that urine can be used to measure the antibody response to a specified microorganism causing infection outside the urinary tract. The possible presence of antibodies in urine should alert researchers who look for antigens in urine to choose assays that minimize interference by such antibodies.
Immunogenic Properties of α(1 → 6) Dextran, Its Protein Conjugates, and Conjugates of Its Breakdown Products in Mice
Authors: MÄKELÄ O. , PÉTERFY F. , OUTSCHOORN I. G. , RICHTER A. W. , SEPPÄLÄ I.
Published: June 2006

Mice were immunized with α(1–6) dextran, either as such or coupled to protein carriers, and their anti‐dextran response was measured by a solid‐phase radioimmunoassay and the Farr assay. Like earlier investigators we found that protein‐conjugated dextran was more antigenic than plain dextran. Our novel findings were that (1) a standard dose (30 μg of dextran per injection) coupled to strongly antigenic protein (chicken serum albumin (CSA)) was three times more antigenic than dextran coupled to weakly antigenic bovine serum albumin (BSA); (2) dextrans of low molecular weight (1000–10,000 daltons) coupled to CSA induced at least ten times stronger secondary responses than did a similarly coupled macromoleculur dextran (5–40 million daltons); (3) variation of the CHO/protein ratio from 0.3 to 1 had little effect on the antigenicity of the dextran. Increase of the ratio from one appeared 10 decrease immunogenicity when BSA was the carrier but not when CSA was ihe carrier.
On the Molecular Basis of T Helper Cell Function
Authors: RUBIN B. , REININGER L. , SUZAN M. , FUERI J. , DENIZOT F. , LANDOLFO S. , AMBLE J. G , LONAI P.
Published: June 2006

B‐lymphocyte promotor factors (B‐LPF) are defined as T‐cell‐derived, released molecules that trigger polyclonal induction of B‐cell differentiation into antibody‐forming cells. B‐LPF activity is independent of antigen, and it apparently induces only IgM‐producing B cells. B‐LPF was discovered as products of an antigen‐specific. I‐Ab‐restricted T‐cell line. We here show that B‐LPF is produced also by lymphoma cells derived from this T‐cell line or by T‐cell hybrids constructed by fusing the T‐cell line with BW5147 thymoma cells. A chicken gamma globulin‐specific T‐cell hybridoma clone also produced B‐LPF. Biological assays demonstrated that B‐LPF‐containing supernatants did not contain IL‐1, IL‐2, B‐cell growth factor, or allogeneic effector factor. Biochemical studies showed that B‐LPF was precipitated by 50% (NH4)2SO4 saturation iind that at least three types of molecules were involved in B‐LPF activity: molecules with molecular weights of >90,000, 50,000–90,000 and 10,000–25,000.The relationship between B‐LPF and antigen‐specific helper/inducer factors is discussed.
Detection of Single Antibody‐Secreting Cells Generated After in Vitro Antigen‐Induced Stimulation of Human Peripheral Blood Lymphocytes
Authors: CZERKINSKY C. , NILSSON L.‐Å. , OUCHTERLONY Ö. , TARKOWSKI A. , GRETZER C.
Published: June 2006

Culture and assay procedures are described for the generation and the subsequent detection of single antigen‐specific antibody‐secreting cells after in vitro stimulation of human peripheral blood lymphocytes with tetanus toxoid. Simple, specific and sensitive, this new assay‐culture system is well suited for the analysis of specific antibody production in man.
Functional Analysis of B‐L (Ia‐Like) Antigen‐Bearing Chicken Peripheral Blood Cells
Authors: HÁLA K. , BOYD R. L. , WOLF H. , BÖCK G. , WICK G.
Published: June 2006

The function of B‐L (Ia‐equivalent)‐positive (B‐L+) and ‐negative (B‐L‐) chicken peripheral blood lymphocytes (PBL) was studied in vitro and in vivo. The PBL were first stained in direct immunofluorescence tests with a fluorescein isothiocyanate‐labelled anti‐B‐L alloantiserum and then separated by means of a fluorescence‐activated cell sorter. In agreement with our previous findings, B‐L‐ cells showed functional properties of T lymphocytes, responding to concanavalin A and phytohaemagglutinin‐P in vitro and inducing a graft‐versus‐host (GVH) reaction when injected into allogeneic embryos. Sorted B‐L+ gave no responses in any of these assays. Neither B‐L+ nor B‐L‐ cells, when tested alone, responded significantly to pokeweed mitogen, but mixtures of the two restored the responsiveness to that of the original unsorted suspension. Of the B‐L+ PBL, 10% were T cells, which may account for the low GVH reactivity given by this population.
Preparative Isolation of Immune Complexes from Serum by Sucrose Gradient Ultracentrifugation
Authors: RØDAHL E. , IVERSEN O.‐J. , DALEN A. B.
Published: June 2006

Immune complexes were isolated by ultracentrifugation in sucrose gradients (20‐65% (w/w)). The centrifugation procedure was demonstrated 10 be isopycnic. The banding density of the complexes was influenced by the chemical nature and molecular size of the antigen and by the antigen to antibody ratio. The method was applied for preparative isolation of immune complexes from patients with systemic lupus erythematosus. rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, and uncomplicated psoriasis.
Fibronectin Promotes Binding But Not Ingestion of Agarose Beads by Mouse Macrophages and Human Monocytes
Authors: GAUPERAA T. , JOHNSON E.
Published: June 2006

We have examined to what extent human fibronectin associated with agarose beads with a 5‐ to 10‐μm diameter mediates binding and uptake of the heads by mouse macrophages and human monocytes. Native agarose beads preincubated with 125I‐fibronectin were neither associated with nor taken up by mouse macrophages after 30 min of incubation under serum‐free conditions. When fibronectin was cross‐linked to cyanogen bromide‐activated agarose heads or incubated with gelatinized heads, this resulted in a significant increase in particle binding by macrophages and monocytes as compared with gelatinized beads, whereas the fraction of cells with ingested particles remained unaltered. Native agarose heads activated by cyanogen bromide and treated with ethanolamine were to a greater extent associated with and taken up by phagocytes than fibronectin‐ or gelatin‐coated heads. Our results indicate thai fibronectin acts as an adhesive glycoprotein and not as an opsonin. Since agarose beads are activators of the alternative pathway of complement, and fibronectin is reported to bind to factor C3, we speculate that cell‐derived C3b is bound to the beads and fibroneetin‐coaled beads arc ingested by the phagocytes via complement C3b receptors on the cells.
Mechanism of Action of Cyclosporin A
Authors: BENDTZEN K. , DINARELLO C. A.
Published: June 2006

Previous studies have shown that cyclosporin A (CyA) prevents the elaboration of the lymphokine leucocyte migration inhibitory factor (LIF). Since LIF production is interleukin 1 (IL‐1)‐dependent, we carried out experiments using partially and highly purified IL‐1 preparations to study the effect of CyA. We found that (a) IL‐1 was consistently depleted during a 1‐h incubation with human blood T lymphocytes but not with B lymphocytes or erythrocytes; (b) the depletion could not be ascribed to pinocytosis, cell functions requiring active metabolism, or enzyme‐mediated destruction of IL‐1; (c) CyA, but not biologically inactive cyelosporin, antagonized the apparent absorption of IL‐1; (d) T cells pre‐exposed to CyA were rendered incapable of removing the monokine; and (e) CyA was capable of displacing IL‐1 once absorbed by T cells. Because the putative binding of IL‐1 showed salurability. reversibility (with CyA as a probe), and tissue specificity consistent with a known target for the monokine, we propose that IL‐1 interacts with a receptor‐like structure on T cells. Finally, we found that insulin interfered with the function of CyA at the very early macrophage‐T‐cell co‐operative stage, even at physiological concentrations.
A Second Component in Bovine AA Amyloid Fibrils Not Identical with Protein AA Is Essential for AA Amyloid Fibrillogenesis
Authors: HOL P. R. , LANGE VELD J.P.M. , van BEUNINGEN‐JANSEN E. W. , VEERKAMP J. H. , GRUYS E.
Published: June 2006

Amyloid fibrils were isolated from the renal papillae and glomeruli of cows with spontaneous AA amyloidosis. The fibrils were solubilized by treatment with guanidine hydrochloride (Gu HCl) and subjected to gel filtration on Sephacryl S‐200. Two other fractions were obtained beside the void volume and the AA fractions. Reaggregation studies were performed by dialysing the fractions, separately or in combinations, against Gu‐HCl‐free solutions. Protein AA alone (about 10 kd) appeared not to precipitate. The olher fractions alone and the combinations of fractions tested formed precipitates. The precipitates containing all fractions (including prolein AA) or protein AA plus a fraction containing a 14‐ and a 23‐kd protein revealed congophilic green birefringent fibrillar material. Dialysis against acidic and calcium‐containing solutions gave the best results. Amyloid fibril‐like material was visible on electron microscopic examination. The amino acid composition of the 19 + 23‐kd material appeared to be slightly different from protein A A and evidently unlike SAP. On imniunolluorescence‐absorbance studies the 19 + 23‐kd material appeared evidently unlike protein A A and SAP. From these findings it is concluded that for spontaneous formation of A A amyloid fibrils other non‐AA proteins are necessary.
On the Interaction between β2‐Microglobulin and Group A Streptococci
Authors: BJÖRCK L. , MIÖRNER H. , KÜHNEMUND O. , KRONVALL G. , SUNDLER R.
Published: June 2006

β2‐microglobulin (β2m) was found to interact with many group A streptococcal strains. The interaction appeared to require multipoint attachment, since monomeric β2m in solution showed no binding, whereas both β2m monomers bound to liposomes, and β2m in aggregates showed affinity for the bacteria. Aggregated HLA antigens (‐A, ‐B and ‐C) and aggregated β2m exhibited the same binding patterns when tested in binding experiments with various group A streplococcal strains. Furthermore, β2m aggregates in excess completely blocked the binding of aggregated HLA antigens, thereby demonstrating that β2m is able to interact with streptococcal surface structures also when it is part of the HLA antigen complex. M protein‐positive group A streptococcal strains bound significantly more β2m than M protein‐negative variants of these strains. Purified M 12 protein partly inhibited the binding of radiolabelled β2‐m aggregates to whole streptococci, and in gel filtration and affinity chromatography experiments, the M 12 protein interacted with β2m. These various data suggest that the interaction between β2m and group A streptococci could be mediated by M protein. Lipoteichoic acid (LTA) is a constituent of the streptococcal cell wall that has been reported to form complexes with M protein at the bacterial cell surface. However, LTA did not influence the interaction between β2m and streptococci, suggesting that the binding of β2m to streptococcal M protein represents a pure protein‐protein interaction. In vivo such an interaction could be established between infecting streptococci and host cells. Among 45 strains of different M types large differences in β2m binding were recorded, whereas among 60 strains of the classical nephritogenic M types 12 and 49, all were highly β2m‐reactive, which points towards a role for β2m in streptococcal pathogenicity.
Analysis of the (H‐2b× H‐2k)F1Restricted Response to Insulin
Authors: RESKE‐KUNZ A. B. , RÜDE E.
Published: June 2006

The aim of these studies was to characterize the (H‐2b× H‐2k)F1‐unique restriction element(s) responsible for presentation of bovine insulin (BI) to a long‐term cultured T‐cell line (BK‐BI‐1.2). (B10.BR × bm12)F1 spleen cells, which express a normal Abα Akβ molecule but a mutated Akα AAbm12β product on their cell surface, were perfectly able to act as BI‐presenting cells. Antibody inhibition experiments with antibodies directed at I‐Ak products revealed that monoclonal antibody 10–2.16, which reacts with the Akβ polypeptide chain, abrogated BI‐directed T‐cell proliferation, whereas antibody H116–32.R5 with specificity for the Akα chain was not inhibitory. These results identified the AbαAkβ complex as restriction structure. Recognition of BI in the context of the Abα Akβ molecule depended on the glutamic acid residue in position 4 of the A chain of bovine insulin. Twenty to twenty‐five percent of the secondary proliferative response of (B10 × B10.BR)F1 lymph node T cells primed with BI in vivo was directed at the A4 determinant, suggesting that BK‐BI‐1.2 T blasts are representative of T‐cell clones with measurable frequency. In (B10.BR × bm12)F1 mice, which lack a functional Abα Abβ restriction element, up to 80% of the proliferative response was dependent on the A4 epitope.
Composition of a Suppressor Factor That Inhibits the Immune Response to Lactate Dehydrogenase B
Authors: IKEZAWA Z. , WALDEN P. , ARDEN B. , NAGY Z. A. , KLEIN J.
Published: June 2006

Hybridomas obtained by fusion of lactute dehydrogenase B (LDHB)‐aciivatcd suppressor T (Ts) cells with the BW5147 thymoma produce a suppressor factor (TsF) that inhibits the proliferation of LDHB‐activated helper T (Th) cells. A similar factor (TsE) is contained in the extract of suppressor hybridomas. Both TsF and TsE are specifically retained by LDHB‐immunoadsorbent columns. Both consist of two components, an antigen‐binding component (ABC) and possibly a major histocompatibility complex (MHC) component. The latter reacts with certain monoclonal antibodies specific for MHC determinants. The two components arc covalently associated in the IsF and noncovalcntly associated in TsE. Mixing of the two components reconstitutes the activity of the TsF or TsE. Disruption of the ABC's tertiary structure results in its inability to reconstitute suppressive activity on mixing with the MHC components. The ABC may contain an intrachain disulphide hond(s). Suppression is obtained when Th cells are incubated first with the ABC and then with the MHC component or vice versa, provided that the incubation period is at least 4 h. The MHC component is also produced by nonsuppressor hybridomas but not by mitogen‐stimulated blasts or by the parental thymoma. The TsF is a glyeoprotein with a molecular weight ot about 120.000 to 160.000. The molecular weight of the ABC is about 76,000–86,000 and of the MHC component about 30,000–37,000.
Lymphoproliferation Induced in Mouse Spleen Cells by Mycoplasma arthritidis Mitogen
Authors: KIRCHNER H. , GIEBLER D. , KEYSSNER K. , NICKLAS W.
Published: June 2006

Spleen cells of various mouse strains (e.g. BALB/c, C3H, and CBA) reacted towards MAS (a mitogen derived from supernatants of cultured Mycoplasma arthritidis) with a marked lymphoproliferative response. This reactivity was T‐cell‐dependent. It was reduced by 90% after removal of macrophages by passage of the spleen cells through Sephadex G‐10 columns. Addition of 2‐mercaptoethanol (2‐ME) to macrophage‐depleted CBA spleen cells completely restored the response to MAS. Spleen cells of C57BL/6 and C57BL/10 mice were unreactive to MAS, even in the presence of macrophages, and this non‐reactivity was controlled by the I‐region of H‐2. Other mouse strains that, similarly to C57BL/6, lack the expression of I‐E on the cell surface (that is, mice of the haplotype H‐2f, H‐2q, and H‐2s) were also non‐responsive to MAS. However, the addition of 2‐ME to spleen cells of non‐responder mice resulted in high lymphoproliferative responses to MAS, which were as high as those of CBA spleen cells. The reaction of C57BL/6 spleen cells to MAS in the presence of 2‐ME again was T‐cell‐dependent, as shown by data with spleen cells of homozygous nude mice and spleen cells treated by anti‐thy‐1 and C. A macrophage dependency of this response was also evident. When C57BL/6 spleen cells were vigorously freed of accessory cells by the use of nylon wool columns, the MAS response could no longer be restored by 2‐ME.
Effects of Interferon‐α/β and Interferon‐γ Preparations on Phagocytosis by Mouse Peritoneal Macrophages
Authors: ROLLAG H. , DEGRÉ M. , SONNENFELD G.
Published: June 2006

The influence of murine alpha/beta‐interferon (Mu IFN‐α/β) and murine gamma‐interferon (Mu IFN‐γ) preparations on the attachment and ingestion phase of phagocytosis by mouse peritoneal macrophages (MPM) was studied. A non‐opsonized strain of Escherichia coli, IgG‐opsonized E. coli, and sheep erythrocytes opsonized with IgG (E‐IgG) and IgM plus complement factor C3b (E‐IgMC) were used as test particles. Pretreatment of MPM with 102‐103 U/ml of Mu IFN‐α/β for 24 h enhanced both attachment and ingestion of bacteria or erythrocytes mediated by the non‐specific receptor, the Fc receptor, or the C3b receptor. Higher concentrations had no such effects. In contrast, treatment of MPM with 101‐102 U/ml of Mu IFN‐γ suppressed attachment and ingestion of non‐opsonized and IgG‐opsonized E. coli and of E‐IgG by 10–40%. Mu IFN‐γ did not influence attachment and ingestion of E‐IgMC. The effects were neutralized by specific anti‐IFN antiserum. The data indicate that the IFN effect on phagocytic activity is, at least to a large extent, due to modifications of the surface receptors.
Induction of Maturation in Different Types of B‐Cell Lymphomas in Vitro with TPA and Antibodies to Surface Immunoglobulin
Authors: BEISKE K. , RUUD E. , MARTON P. F. , GODAL T.
Published: June 2006

Cells from eight selected cases of human non‐Hodgkin lymphomas of various histological types (lymphocytic, centrocytic, centrocytic/centroblastic, and immunocytomas) were stimulated in vitro with 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) and anti‐immunoglobulins (anti‐Ig) against the surface immunoglobulin (sIg) on the tumour cells. Six of these cases responded by intracellular Ig accumulation as measured by flow cytofluorometry and direct phenotypical change into immunoblasts/plasmablasts as detected by light microscopic immunocytochemistry. However, the response to TPA alone varied considerably from ease to case. These findings suggest that many, if not all, B‐cell subsets have the capacity to develop directly into Ig‐synthesizing cells (immunoblasts and plasmablasts). However, conditions for eliciting such events may vary, depending on the phenotypical properties and differentiation stage.
Human Amyloid P Component: An Elastase Inhibitor
Authors: LI J. J. , McADAM K. P. W. J.
Published: June 2006

Amyloid deposits almost invariably contain a pentagonal‐shaped protein (a so‐called pentraxin), termed amyloid P component (AP), in close apposition to the amyloid fibrils. AP is also detected alongside normal dust elastin fibres in skin and basement membrane. In the present studies, purified human AP was shown to inhibit the activity of porcine pancreatic elastase. The inhibition of elastolytic activity was not abolished by heating AP to 70°C. Furthermore, two other human serum proteins used as controls did not inhibit elastase activity: albumin, which has a similarly acidic pI, and C‐reactive protein, which is a pentraxin, sharing 55% sequence homology with AP. Enzyme kinetic studies showed that elastase treated with AP had a slower elastolytic rate than untreated elastase. The inhibitory effect of AP was reversed by high substrate (fivefold) concentration. These observations suggest that AP may function in vivo to protect elastin and amyloid fibrils from proteolytic cleavage, Indeed, this may in part account for the relative resistance of amyloid deposits to resorption and proteolysis.
Delayed‐Type Hypersensitivity after Immunization with Ultrasonicated Mycobacterium lepraemurium in C3H and C57BL Mice
Authors: LØVIK M. , HAUGEN O. A. , CLOSS O.
Published: June 2006

Subcutaneous immunization with ultrasonicated Mycobacterium lepraemurium (MLMSon) in incomplete Freund's adjuvant (IFA) induced long‐lasting skin reactivity with the kinetics of a tuberculin‐type delayed hypersensitivity (DTH) reaction in both C3H and C57BL mice. The reactivity generally was stronger in C57BL than in C3H mice, and with increasing doses of MLMSon test antigen the local reaction increased more in C57BL than in C3H mice. Pretreatment of C3H mice with cyclophosphamide before immunization caused a shift in the dose‐response curve so that the local reaction increased more with increasing doses of test antigen. Histological examination of the reaction elicited by MLMSon in immunized mice revealed a predominantly mononuclear cell infiltrate, and local reactivity could be transferred by immune cells but not by immune serum. The local reaction elicited by MLMSon exerted an adjuvant effect on the induction of DTH to sheep erythrocytes. Thus, MLMSon in IFA given subcutaneously induced stable DTH that conformed to the criteria for tuberculin‐type DTH.
Effect of Bestatin on in Vitro Responses of Murine Lymphocytes to T‐Cell Stimuli
Authors: DUNLAP B. E. , DUNLAP S. A. , RICH D. H.
Published: June 2006

Bestatin, a dipeptide protease inhibitor that has been reported to have immunomodulatory properties, was tested for its ability to augment the in vitro response of murine lymphocytes to allogeneic cells and to concanavalin A (Con A) or NaIO4 treatment. Bestatin augmented the response of spleen cells to allogeneic stimulation but was inhibitory in Con A and NaIO4 mitogenic assays. No augmentation of the allogeneic response was found if cells from responder and stimulator strains were genetically similar for the I region of the major histocompatibility complex. Bestatin did not restore the reduced mitogenic response of spleen cells from cyclophosphamide‐treated animals. Although bestatin had no influence on interleukin activity once produced, it enhanced the production of interleukin activity in Con A cultures.
Subclass Distribution of Human Anti‐Staphylococcus aureus Alpha Toxin Antibodies: Suggestion of an IgG1, IgA1, IgG4 Switch Pattern
Authors: HAMMARSTRÖM L. , PERSSON M. A. A. , SMITH C. I. E.
Published: June 2006

The subclass distribution of antibodies against alpha toxin from Staphylococcus aureus in man was investigated. Antibodies were restricted to IgG1, IgA1, and IgG4 and appeared to develop sequentially. These data suggest a distinct pattern of class and subclass switches in response to protein antigens. However, studies in immunodeficient (class‐ or subclass‐ deficient) donors show that the proposed patterns is not obligatory.
The MT4 Allodeterminant Is Borne on an HLA‐DS Molecule on DR5 Cells
Authors: ZANGARA R. J. , KANNAPELL C. C. , KARR R. W. , GOYERT S. , SILVER J. , SCHWARTZ B. D.
Published: June 2006

Human HLA‐DR molecules have been shown to be structurally homologous to the murine I‐E subregion molecules by amino acid sequence analysis. Recent studies have demonstrated the isolation of an I‐A subregion‐homologous molecule (HLA‐DS) from human B‐cell lines with the rabbit antiserum RbO3, made against a marmoset I‐A‐like Ia molecule. Previous work from our laboratory has demonstrated that the DR5 homozygous lymphoblastoid cell line Swei expresses at least two different Ia α chains and four different Ia β chains, which associate to form four distinct human Ia molecules, α1β2, α1β3, α2β1, and α2β4, and that the α2β1 molecule bears the allodeterminants MB3 and MT4. To determine whether the MT4‐bearing α2β1 molecule was an HLA‐DS molecule, the α2β1 molecule reactive with an anti‐MT4 alloserum was compared with the Ia molecule reactive with the rabbit xenoantiserum RbO3 by two‐dimensional gel electrophoresis and sequential immunoprecipitation. The α2 chain and the RbO3‐reactive α chain yielded essentially similar spot patterns. The β1‐chain spot pattern was a subset of the RbO3‐reactive β‐chain spot pattern. Sequential immunoprecipitation indicated that RbO3 removed all molecules reactive with MGH88B. These results indicate that on DR5 cells the allosera‐reactive α2β1 molecule, which bears MT4, is an HLA‐DS molecule.
Immune Characteristics of the Beige‐Nude Mouse
Authors: FODSTAD Ø. , HANSEN C. T. , CANNON G. B. , BOYD M. R.
Published: June 2006

Double immunodeficient mice, homozygous for both the beige and the nude genes, were developed on C57B1/6N‐beige and N :NIH(S)‐nude background through five steps of mating. The animals were healthy and had a life span comparable to that of regular nude mice. Beige and regular nude mice showed a significant difference in natural killer (NK) activity, with mean±SE values of 6±1.0% and 25±2.8%, respectively. The response to the T ‐cell mitogens phytohaemagglutinin and concanavalin A was similar in beige and NIH mice of the same genotypes for the nude genes. Both groups of nu/nu mice also showed some response in these assays, as well as in the plaque‐forming cell assay, indicating that nude mice are not completely devoid of functional T or T‐like cells. The nude genes were found to increase the response to the B‐cell mitogen lipopolysaccharide in both the beige and NIH animals, whereas the response was reduced in mice of the beige genotypes. The viable, low NK beige‐nude mice here reported on may be a valuable tool for studying factors involved in host defence against tumours. Interestingly, no spontaneous tumours have so far been observed in such mice.
Heterogeneity in Expression of the T4 Epitope in Black Individuals
Authors: STOHL W. , KUNKEL H. G.
Published: June 2006

The T‐cell differentiation antigen T4/Leu3 has been described as a non‐polymorphic molecule important in T‐cell recognition of class II major histocompatibility complex antigens. We report the polymorphism of this molecule in black individuals as manifest by a heterogeneity of staining with OKT4 monoclonal antibody. No such heterogeneity was observed when staining with other monoclonal antibodies that bind to different epitopes of the same molecule. No heterogeneity of staining with any of the monoclonal antibodies was observed in whites. Three patterns of OKT4 staining emerged: intact, deficient, and intermediate. This heterogeneity is likely to be due to an intrinsic heterogeneity in T4 epitope expression and not secondary to an interfering plasma factor as shown by the preservation of the T4 epitope pattern after a 3‐day culture in the presence or absence of mitogen. Family studies strongly suggest that this heterogeneity in T4 epitope expression is inherited in an autosomal codominant fashion.
Secondary Immunization with a Protein Antigen (Tetanus Toxoid) in Man
Authors: DONNENBERG A. D. , ELFENBEIN G. J. , SANTOS G. W.
Published: June 2006

Tetanus toxoid (TET) was used as an immunogen to explore regulatory events occurring after secondary immunization in man. Changes in dose‐response patterns, kinetics, and frequency of antigen‐ reactive cells in the peripheral blood, and the serum antibody titres were studied. The most striking feature of these studies was the finding that immediately after immunization, a brief period of decreased responsiveness preceded the expected amplification of antigen‐specific humoral and cell‐mediated responses. One day after immunization serum antibody levels declined to approximately half their initial values before the expected increase in titre These results could not be explained by the formation of antigen‐antibody complexes. Lymphoproliferative responses were also depressed 1 day after immunization but increased in both magnitude and sensitivity from 7 to 28 days after challenge. A decrease in thymidine incorporation evident at supraoptimal antigen concentrations also became progressively more prominent during this interval Cell mixing experiments documented the presence of a radioresistant population present 1 day after immunization capable of suppressing TET‐specific lymphoproliferative responses of autologous lymphocytes obtained before immunization. These suppressors are preferentially activated at high antigen concentrations. Limiting dilution analysis revealed a 30‐fold increase in TET‐reactive proliferating cells by 28 days after immunization. Expansion occurred first in cells recognizing high antigen concentrations and subsequently in cells rccognizing lower doses. Taken together, these findings present a consistent if somewhat counterintuitive picture of some of the regulatory processes accompanying booster immunization. The earliest detectable event appears to be the application of an active suppressor mechanism, release from which signals initiation of those processes classically associated with an anamnestic response
Factor I Deficiency and C3 Nephritic Factor: Immunochemical Findings and Association with Neisseria meningitidis Infection in Two Patients
Authors: TEISNER B. , BRANDSLUND I. , FOLKERSEN J. , RASMUSSEN J. M. , POULSEN L. O. , SVEHAG S.‐E.
Published: June 2006

The complement system was examined in two patients with systemic Neisseria meningitidis infections, both of whom had reduced or nondeteclable CH50 as analysed by both pathways. C3 measured by conventional technique revealed 19% anti‐C3c‐reactive protein in the plasma of patient 1 and 3% in patient 2. Patient 1 had circulating C3b but no detectable C3c, C3d, or C4d, whereas patient 2 had normal levels of C3c and C4d and strongly elevated levels of C3d. Factor B analysis revealed no demonstrable native factor B and small amounts of Bb in patient 1 and normal concentration of native factor B plus trace amounts of Bb in patient 2. The depletion of C3 in both patients was due to uncontrolled activation caused by complete factor I deficiency (patient 1) and circulating C3 nephritic factor (patient 2). Both parents of patient 1 had factor I concentrations below (mean‐2 SD) that seen in normal healthy individuals (n= 20). Circulating immune complexes (IC) were demonstrated in patient 1 only, whereas serum from both patients had strongly reduced capacity to solubilize preformed IC.
The Effect of Monoclonal Antibodies against Escherichia coli Type 1 Pili and Capsular Polysaccharides on the Interaction between Bacteria and Human Granulocytes
Authors: SÖDERSTRÖM T. , ÖHMAN L.
Published: June 2006

Monoclonal antibodies against Escherichia coli type 1 pili and the K13 capsular polysaccharide strongly influenced the interaction between human polymorphonuclear leucocytes (PMNL) and E. coli 06:K13:H1. Bacteria with type 1 pili, associated with the neutrophils, caused a metabolic activation but were not ingested. Addition of monoclonal antibodies to type 1 pili resulted in increased association, but also phagocytosis and metabolic activation of the granulocytes. Monoclonal antibodies against the Kl3 polysaccharide strongly stimulated phagocytosis, especially of the type 1 piliated bacteria, suggesting a synergistic effect between binding of type 1 pili and the Fc part of the capsular antibodies to the PMNL. Addition of D‐mannose inhibited the opsonization of type 1 piliated E. coli 06:K13:H1 in the presence of both type 1 pilus antibodies and capsular antibodies. Monoclonal anti‐idiotypic (anti‐anti‐capsular) antibodies reduced the association with the PMNL of the bacteria preopsonized with anti‐capsular antibodies. The bacterial ingestion and the metabolic activation of the PMNL were also reduced, suggesting a role for anti‐idiotypic antibodies in specific modulation of inflammation.
Purification of Cytostatic Protein Factors Released from Human Monocytes
Authors: NISSEN‐MEYER J. , KILDAHL‐ANDERSEN O.
Published: June 2006

Human monocyles release cytostatic protein factors (CF) upon activation with lymphokines und lipopolysacchairide. CF has been purified from monocyte supernatants by ion‐exchange chromatography. chromatofocusing and gel filtration, and this resulted in a 10,000‐fold reduction in the amount of protein in the purified CF preparation compared to the amount in the monocyte supernatant. About 5% of the eytostatic activity was recovered after purification. CF constitutes a population of proteins heterogeneous with respect to ionic charge and differing in their isoelectric points, since CF eluted as a broad peak both upon ion‐exchange chromatography and chromatofocusing. The isoelectric points of the CF proteins were determined by chromatofocusing to be between 6.0 and 5.0. The molecular weight of CF as determined by gel filtration was in the range of 45,000 to 35,000 daltons. Only one monocyte‐secreted protein with a molecular weight of 40,000 was detected upon sodium dodecyl sulphate‐polyacrylamide gel electrophoresis of proteins in the purified CF preparations obtained after the final gel filtration step, und this protein may consequently be CF. If the 40,000‐dalton protein is CF. one may estimate that about 106 monocytes produced roughly 0.1 μg CF upon activation and that significant cytostasis may be detected with less than nanogram quantities of CF.
Characteristics of the Strongly Ia‐Positive Cells in Rat Liver
Authors: LAUTENSCHLAGER I.
Published: June 2006

The expression of MHC class II antigens (Ia) on rat liver non‐hepatocytic cells was analysed by the Staphylococcus aureus rosette method using monoclonal antiserum to the common part of the class II molecules. Of the non‐hepatocytic liver cell population, the passenger lymphocytes and monocytes were strongly anti‐Ia binding but the most reactive were large mononuclear cells, morphologically macrophages. The characteristics of the strongly Ia‐positive cells of rat liver were tested with different‘macrophage markers'. Of the macrophage‐like cells, 55% were Ia‐positive in the Staph. aureus rosette 84% had intracytoplasmic lysozyme, 100% of them were positive for α‐naphthyl acetate esterase (ANAE) reaction, and 92% had Fc‐receptors on the cell surface. As functional tests of macrophages, phagocytosis and adhesion were used: 79% of the cells were able to phagocytose antibody‐coated human red cells and 56% of the cells adhered on glass. As most of the tested cells were positive for the markers the results demonstrate that the most antigenic component of rat liver has the characteristics of macrophages and thus the strongly Ia‐positive cells are probably liver tissue macrophages (Kupffer cells)
Concomitant Induction and Persistence of Hapten‐Specific Suppressor and Helper T Cells in Vivo
Authors: ZOLLER M. , WIGZELL H. , ANDRIGHETTO G.
Published: June 2006

Intravenous injection of haptenized syngeneic lymphoid cells in the mouse induced a suppression in vivo against any immunogen carrying this hapten, introduced as a second antigen. Suppression was observed against any epitope on such a haptenized immunogen, thus largely excluding cross‐reactions at the level of antigen‐binding or idiotypy. However, when cells from such suppressed mice were assessed in vitro, it could be shown that significant T‐helper activity had been induced by the same procedure, which in vivo resulted in suppression only. Thus, concomitant induction and persistence of hapten‐specific suppressor and helper T cells is a result of the present immunization protocol. Both phenomena express the conventional requirements for physical linkage between hapten and immunogen to have an impact on the antibody response against the epitopes of the carrier. It is thus likely that the observed suppression/help in the present system does function at the level of handling the intact immunogen.
Detection of α1 and α2 Heavy‐Chain Constant‐Region Genes in Common Variable Hypogammaglobulinaemia Patients with Undetectable IgA
Authors: SMITH C. I. E. , HAMMARSTRÖOUM L.
Published: June 2006

Five patients with common variable hypogammaglobulinaemia with undetectable levels of IgA in serum and saliva were investigated for immunoglobulin structural gene defects. All five patients carried in their genome α1 and α2 genes as detected in Southern blotting analysis by means of a cloned Cα gene probe.
Lymphoproliferative Responses of Spleen Cells of Inbred Rat Strains to Mycoplasma arthritidis Mitogen
Authors: MORITZ T. , GIEBLER D. , GÜNTHER E. , NICKLAS W. , KIRCHNER H.
Published: June 2006

A potent mitogen for T lymphocytes of various species has recently been isolated from the supernatant of cultured Mycoplasma arthritidis organisms (MAS). In the mouse, reactivity to this mitogen has been observed to be controlled by the I‐E subregion of the major histocompatibility complex. We have analysed the responses of spleen cells from several inbred rat strains covering practically all known haplotypes of the major histocompatibility complex of the rat (RT1). Unlike in the mouse, all of these responded well to MAS, except for the BN rat strain, which is a low responder to all T‐cell mitogens, including phytohaemagglutinin and concanavalin A. This unresponsiveness, however, appeared to be unrelated to the RT1 haplotype, since LEW.1N rats carrying the same RT1u haplotype as BN animals responded well. Mice of the strain C57BL/6 are non‐responders to MAS, but—as previously shown—their spleen cell responses can be reconstituted by the addition of 2‐mercaptoethanol. No such reconstitution was observed for the low responsiveness of BN rat spleen cells. Stimulation with MAS induced high titres of interferon (presumably gamma interferon) in spleen cells from all rat strains tested. Spleen cells from BN rats produced lower interferon activities than those from other strains.
Auto‐Delayed‐Type Hypersensitivity Induced in Immunodeficient Mice with Modified Self‐Antigens
Authors: TARCIC N. , KLEIN B. Y. , NAOR D.
Published: June 2006

Suppressor cells obtained from spleens of normal A mice, or factor extracted from these suppressor cells, abolished the syngeneic delayed‐type hypersensitivity (syn‐DTH) response of X‐irradiated A mice injected with trinitrophenylated spleen cells und challenged with syngeneic lymphoblasts. Some of the physical, chemical and biological properties of the suppressive factor (SF) were characterized. The SF was relatively temperature‐stable and its activity was destroyed by pronase (but not with RNase or DNase). The activity of the SF was absorbed on concanavalin A and anti‐I‐Jk Scpharose columns, suggesting that the factor is a glycoprotein‐bearing I‐Jk product. The approximate molecular weight of the factor is 50,000–60,000. The SF was absorbed on plastic adherent cells (but not on non‐adherent cells). Adherent cells that absorbed the SF abrogated the ability of primed T cells to transfer the syn‐DTH to naive X‐irradiated recipients. In contrast, SF that was presented directly to the primed T cells failed to abolish their ability to transfer DTH. These findings suggest that the adherent cells serve as mediators, transferring the SF from factor‐producing cells (Lyt‐1+2+3+, I‐Jk+ T cells) to target cells (Lyt‐1+ primed T cells).
Antibody Responses to λ1J558 and λ2315 Light Chains
Authors: BOGEN B.
Published: June 2006

Specificity of BALB/c antibody responses to A chains of isologous myeloma proteins 315 and J558 was explored by enzyme‐linked immunosorbent assay. A‐chain binding antibodies were not detected when immunizing with assembled (H + L) myeloma proteins. However, relatively high titred IgG antibodies were elicited by free λ2315 immunization. Antibodies were directed to ‘hidden’ determinants since binding was abrogated upon H + L assembly of chains. At least a portion of antibodies bound antigenic determinants in the variable region and cross‐reacted with λl land λ3 chains. Free λ11558 immunization induced low‐titred, predominantly IgM antibodies that also only reacted with ‘hidden’ determinants. These determinants were most probably located in the constant (C) region and no cross‐reaction to λ2 or λ3 was observed. An artefact of technical importance was noted: myeloma proteins exposed ‘hidden’ determinants on their A chains when coated directly to polystyrene walls. This ariefactual exposition was lost when anti C‐region antibody spacer molecules were inserted between the wall and the myeloma proteins. Antibody and T helper cell (Th) responses to free λ2315 covaried significantly in various strains while antibody and Th responses to free λ11558 did not. In some strains, weak antibody responses were detected without detectable Th.
Secretion of γ‐Interferon at the Cellular Level
Authors: ANDERSSON U. , MARTINEZ‐MAZA O. , ANDERSSON J. , BRITTON S. , GAOLER H. , DE LEY M. , MODROW S.
Published: June 2006

Using a haemolytic plaque assay for y‐interferon (IFN‐γ) secretion we found that in vitro Epstein‐Barr virus (EBV) exposure of peripheral blood mononuclear cells from EBV immune individuals led to IFN‐y secretion, which was apparent wiihin 6 h after virus coniact and peaked 12–24 h after induction. Live, ultraviolel‐light‐irradiaicd and heat‐inactivated virions all caused IFN‐γ secretion. In contrast, blood mononuclear cells from EBV non‐immune adults or neonates could not be activated to IFN‐y production by EBV.
Monoclonal Antibodies to a Purified Human Transferrin Receptor
Authors: NIKINMAA B. , ENNS C. A. , TONIK S. E. , SUSSMAN H. H. , SCHRÖDER J.
Published: June 2006

BALB/c mice were injected with an affinity‐chromatography‐purified placental transferrin receptor preparation. Spleen cells were fused with NS‐1 myeloma cells. Sixteen hybrids producing monoclonal antibodies specific for the transferrin receptor and two hybrids specific for transferrin were identified by radioimmunoassay (RIA). Five hybrids were selected for cloning on the basis of antibody specificity and affinity. None of the antibodies inhibited the binding of transferrin to K562 cells. The binding of antibody ID9 to K562 cells was partially inhibited by transferrin or a polyclonal goat anti‐transferrin receptor antiserum. Of the five antibodies, two (IIB6 and IIB2) reacted only with the purified receptor and solubilized cells and not with whole cells. The other three antibodies, when tested with normal human cells and leukaemia and tumour cell lines, showed identical reaction patterns. The antibodies precipitated a glycoprotein from K562 cells with an apparent molecular weight of 94,000, estimated from sodium dodecyl sulphate‐polyacrylamide gel electrophoretograms run under reducing conditions, and a molecular weight of 188,000 when run under unreduced conditions. All antibodies have a high affinity with Ka values ranging from 1.44 ± 109 to 3.56 ± 1010 (1/mol). The antigen precipitated by all five antibodies showed identical peptide maps after partial proteolytic digestion.
The Role of Viral Glycoproteins in Mumps Virus‐Mediated Antibody‐Dependent Cellular Cytotoxity in Vitro
Authors: ALSHEIKHLY A.‐R. , ORVELL C. , WØHLIN B. , ANDERSSON T. , PERLMANN P.
Published: June 2006

Treatment of human peripheral blood lymphocytes with live or UV‐inactivated mumps virions enhances antibody‐mediated cellular cytotoxicity (ADCC), reflected by increased target cell lysis in a 51Cr‐release assay or an increased number of plaque‐forming cells on monolayers of bovine erythrocytes (Eb) in the presence of anti‐Eb antibodies. Virus treatment of the Eb targets causes a similar enhancement. The role of viral glycoproteins in ADCC enhancement was investigated by using a panel of monoclonal antibodies raised in mice against mumps virions. Most of the lymphocytes bound mumps virions, as ascertained by indirect immunofluorescence. A high proportion of virus‐treated lymphocytes also formed rosettes with Eb. Anti‐HN antibodies inhibited resetting to various degrees. Although antibodies with high haemagglutination inhibition titres were most efficient inhibitors, antibodies without this serological activity were also inhibitory. Anti‐F antibodies were only weakly inhibitory, and anti‐NP antibodies had no effect. Anti‐HN antibodies also abrogated target cell lysis in the 51Cr‐release assay and effector cell recruitment in the ADCC plaque assay by inhibiting virus‐mediated Eb‐lymphocyte interactions both at the target cell and at the effector cell level. Anti‐F or anti‐NP antibodies were only weakly or not at all inhibitory. The results suggest that virus‐mediated enhancement of ADCC is caused by the HN glycoprotein, primarily (although perhaps not exclusively) by its improvement of the effector cell‐target cell contacts necessary for the efficient execution of target cell lysis.
Effect of 5′‐Methylthioadenosine, 3‐Deazaadenosine, and Related Compounds on Human Natural Killer Cell Activity
Authors: FREDHOLM B. B. , JONDAL M. , LANEFELT F. , NG J.
Published: June 2006

The effect of 5′‐methylthioadenosine (MTA) on human natural killer (NK) cell activity was examined and compared with the effect of 3‐deazaadenosine (c3‐ado) and periodate‐oxidized adenosine (ado‐ox). MTA inhibited NK cell activity in concentrations above 30 μM, but in concentrations below 10 μM a slight enhancing effect was often observed. C3‐ado and ado‐ox were 10 and 3 times more potent, respectively, as inhibitory agents and did not increase NK cell activity in low concentrations. The inhibitory effect of c3‐ado was unaffected by preincubation of the cells but was enhanced by the addition of μ‐homocysteine. In concentrations that caused inhibition of NK cell activity all three agents caused a fall in the methylation index (AdoMet/AdoHcy) but no or an inconsistent effect on the level of cyclic AMP. An increase in the level of AdoHcy was observed already after 1 μ of incubation but was more pronounced after 4 μ of preincubation with the adenosine derivatives. The inhibition of cytotoxicity was mainly on their initiation of lysis, with a smaller effect on target cell binding Antibody‐dependent cellular cytotoxicity and lectin‐dependent cellular cytotoxicity appeared to be less sensitive to inhibition by c3‐ado. Our results show that several adenosinc analogues inhibit NK‐cell‐mediated cytotoxicity in parallel with a decreased methylation index. The results suggest that a methylation step is critical in lymphocyte‐mediated cytotoxicity and that NK cell activity is more sensitive to inhibition of this step than antibody‐ or lectin‐dependent cytotoxicity.
In Vitro Induction of Specific T Cells Able to Help in the Generation of Delayed‐Type Hypersensitivity by Thymus‐Dependent and Type‐II ‘Thymus‐Independent’ Antigens
Authors: BRETSCHER P. A.
Published: June 2006

An in vitro system is described which supports the primary induction of T cells able to help in the induction of delayed‐type hypersensitivity. This system is used to demonstrate that hapten‐Ficoll conjugates induce potent hapten‐specific T cells with this function. These type‐II ‘thymus‐independent’ antigens are thus able to induce specific regulatory T cells. These helper T cells are similar to those induced by classical thymus‐dependent antigens in that they are specific for antigen rather than idiotype and act by a linked mechanism.