bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of Monoclonal Antihuman‐B‐Cell Antibody BL13 as an Anti‐C3d‐Receptor (CR2) Antibody
Authors: COHEN J. H. M. , FISCHER E. , KAZATCHKINE M. D , BROCHIER J , REVILLARD J. P.
Published: June 2006

BL13, a mouse monoclonal IgGl antibody raised against human B cells, blocked the function of the C3d receptor (CR2) and bound with high affinity (5x 108 L M−1) to CR2 on B lymphoma cells. Following capping with the second antibody BL13 inhibited C3d‐dependent rosette formation of Daudi and Raji cells and C3b‐dependent CR2‐mediated rosette formation with B lymphoma cells, but did not inhibit CRI‐mediated rosettes between C3b‐bearing cells and peripheral blood lymphocytes. Competitive binding experiments between biotinylated BL 13 or anti‐CR2 antibody HB‐5 and unlabelled antibodies demonstrated that BL13 bound to an epitope that is distinct from that recognized In HB‐5, and closely associated with that recognized by monoclonal antibody ann‐B2, BL13 only reacted with some B cells and follicular dendritic cells in germinal centres in human lymph nodes, whereas HB‐5 strongly reacted with circulating B cells and bound to most cells in the follicles. These results demonstrate the heterogeneity of antigenically defined CR2.
Genetic Variation in the in Vitro Veto Activity of Bone‐Marrow Cells
Authors: HURME M.
Published: June 2006

Cells exerting a veto type of suppressive activity, that is, suppressing the cytotoxic T lymphocyte formation against the class I major histocompatibility complex antigen on their surface, can be found in different lymphoid tissues, for example, in the bone marrow. In this study we show that there is variation in the in vitro veto‐cell activity between bone marrow cells derived from different mouse strains: bone marrow cells derived from mice carrying the non‐H‐2 genes of B10/B6 mice had clearly the strongest veto activity.
The Sheep Erythrocyte Receptor and Both α and β Chains of the Human T‐Lymphocyte Antigen Receptor Bind the Mitogenic Lectin (Phytohaemagglutinin) from Phaseolus vulgaris
Authors: LECA G. , BOUMSELL L. , FABBI M. , REINHERZ E. L. , KANELLOPOULOS J. M.
Published: June 2006

We have studied the interaction of mitogenic lectins such as phytohaemagglutinin (PHA) and concanavalin A (Con A) with both surface molecules which, by the use of monoclonal antibodies, are known to trigger T‐cell mitogenesis. Monoclonal antibodies recognizing the T‐lymphocyte receptor for antigen (Ti) and/or its associated structure, CD3, activate T cells. More recently, a second pathway of activation has been described which involves the sheep erythrocyte binding glycoprotein CD2. a surface molecule distinct from Ti‐CD3. Lysates from surface‐iodinated T‐leukaemia cell lines were treated with lectin and affinity purified anti‐lectin antibodies coupled to protein A‐Sepharose. We have shown that eluatcs from Con A/anti‐Con A or PHA/anti‐PHA immunoprecipitates contained Ti. since;a rabbit anti‐Tα serum, which recognises the native and denatured forms of the constant region of the α chain, immuno‐precipitated Ti from these eluates. Furthermore, Ti immunoprecipitalcd bv anti‐Tα serum from lysates of surface iodinated E lymphocvtes was binding to PHA after elution from the immunoprecipitate. When the purified Ti molecule was reduced and alkylated, allowing the permanent dissociation of its α and β subunits, PHA interacted with both chains, whereas anti‐Tα serum immunoprecipitalcd the α chain onlv. Altogether, these results demonstrate that PHA interacts with both chains of the T cell receptor for antigen on human peripheral T lymphocytes. With the HPB‐ALL tumour line, a similar approach showed that both α and β chains of Ti bind to Con A and Ulex europtaeus‐ 1 but not Helix pomatia. Affinity chomatographv on immobilized lectins and immunoprecipitation with lectin/anti‐leelin antibodies were employed to test whether CD2 binds to PHA and Con A. The results show that CD2 from human peripheral T lymphoevtes binds both lectins but with a lower affinity for PHA than Con A.
Enhancement of Immunoglobulin Secretion by the Lymphokine‐Like Activity of a Mycoplasma arginini Strain
Authors: RUUTH E. , LUNDGREN E.
Published: June 2006

B lymphocytes, preactivated by lipopolysaccharide (LPS), could he triggered to growth by a strain of Mycoplasma arginini, while the level of immunoglobulin (Ig) secretion, quantitatesd as the number of plaque‐forming cells (PFC). was low. The PFC response could be increased by the addition of conditioned media (CM) from lectin‐activated spleen cells or T cell tumour EL.‐4 to the culture of preactivated B cells. CM did not by itself induce a significant amount of PFC in the cultures of LPS‐preactivated B cells. The maturation enhancing activity was distinct from 1L‐2 and B cell growth factor as judged by gel exclusion chromatography.
Immunological Studies in the Acquired Immunodeficiency Syndrome
Authors: HOFMANN B. , ØDUM N. , JAKOBSEN B. K. , PLATZ P. , RYDER L. P. , NIELSEN J. O. , GERSTOFT J. , SVEJGAARD A.
Published: June 2006

The lymphocyte transformation responses to mitogens (phytohaemagglutinin (PHA), concanavalin A (Con A), and pokeweed mitogen (PWM)), allogeneic cells, and the antigen‐purified protein derivative (PPD) were studied in six acquired immunodeficiency syndrome (AIDS) patients and in six healthy controls, each of whom was HLA‐DR‐ and mixed lymphocyte culture (MLC)‐identical with one of the AIDS patients. No evidence of suppression was observed when irradiated or non‐irradiated AIDS peripheral blood mononuclear cells (PBMC) were added to cultures of HLA‐DR‐identical PMBC from healthy controls stimulated with the strong mitogens PHA and Con A or with allogeneic cells, but suppression may be involved in the decreased responses in cultures stimulated with PWM or PPD. Addition of supernatants from macrocultures of AIDS cells did not suppress responses of control PBMC. Thus, suppression by any lymphocyte subset or soluble factor alone cannot explain the generally severely depressed transformation responses in AIDS. Addition of heavily irradiated HLA‐DR‐identical PBMC from healthy controls or supernatants from these cultures led to increased responses in cultures of mitogen‐stimulated AIDS PBMC and in some cultures of antigen or allogeneic cell‐stimulated AIDS PBMC, which were of the same magnitude as seen after the addition of commercially obtained T‐cell growth factor (TCGF). This indicates that AIDS cells are deficient in producing TCGF.

Heavily irradiated AIDS PBMC were capable of restoring the transformation responses to mitogens and antigens of purified HLA‐DR‐identical normal T cells, indicating that AIDS cells have a normal antigen‐presenting capacity and interleukin (IL‐1) production. However, AIDS PBMC had a very poor capacity to stimulate normal PBMC in MLC. Together, our experiments suggest that the immune deficiency in AIDS cells may be partially due to a decreased capability of T lymphocytes to produce TCGF and that a decreased number and/or function of dendritic cells may also be involved.
Cell‐Mediated Immune Response in the Diseased Joints in Patients with Reactive Arthritis
Authors: KONTTINEN Y. T. , NORDSTRÖM D. , BERGROTH V. , LEIRISALO‐REPO M. , SKRIFVARS B.
Published: June 2006

To evaluate the level of lymphocyte activation in reactive and rheumatoid arthritis, density gradient‐isolated, synovial fluid mononuclear cells were stained with a panel of antisera directed at lymphocyte activation markers using an avidin‐biotin‐peroxidase complex (ABC) method. More specifically, we studied the expression of immune response‐associated class II HLA antigen (la), of receptors for interleukin 2 (Tac) and transferrin (T9), as well as of gp 40/80 glycoprotein (4F2). Although Ia+ cells formed about 60% of all the synovial fluid mononuclear cells in both disease conditions, the proportion of Tac+ (33±4% vs 3±1%, P<0.001), T9+ (34±4% vs 5±2%, P<0.001), and 4F2+ (48±6% vs 3±2%, f<0.001) cells was high only in reactive arthritis. All the patients who had reactive arthritis followed a favourable clinical course during the 4‐month‐long prospective follow‐up, whereas disease activity was stable in patients with rheumatoid arthritis. These findings suggest that the diseased joints in reactive arthritis are a site for an active, but normally down‐regulated, cell‐mediated immune response.
Synergistic Action of Gamma Interferon and Catalase to Reverse the Suppressive Effect of Peritoneal Macrophages on Concanavalin A‐Induced Lymphocyte Proliferation
Authors: DOHLSTEN M. , LINDÉN O. , HEDLUND G. , SJÖGREN H. O. , DIAMANTSTEIN T. , CARLSSON R.
Published: June 2006

The suppressive effect of normal rat peritoneal exudate cells (PEC) on concanavalin A (Con‐A)‐induced lymphocyte proliferation was studied. Partial suppression of proliferation was obtained by adding 3% PEC and complete suppression wits observed with 6% PEC. The suppressive effect was mediated by W3/25 plastic‐adherent macrophages, which constitute about 60 % of normal PEC. Addition of PEC prior to. simultaneously with, or 24 h after, but not 48 h after, the stimulation of lymphocytes with Con A resulted in suppression. Suppressed cultures produced normal or slightly increased amounts of interleukin 2 (IL‐2). but the expression of the IL‐2 receptor on lymphocytes was decreased. Pre‐exposure of PEC to gamma interferon (IFN‐γ) resulted in decreased suppression, whereas IFN‐γ added simultaneously with the lymphocytes had no effect. Catalase reversed PEC‐induecd suppression and significant synergistic effects were recorded when combined with IFN‐γ. Even completely suppressed cultures were effectively protected from suppression. Indomethacin and combinations of indomethacin with catalase or IFN‐γ did not result in additional protection Irons PEC‐mediated suppression.
The Interaction between Cell‐Surface Antigens and Antibodies Bound to Monodisperse Polymer Particles in Normal and Malignant Cells
Authors: DANIELSEN H. , FUNDERUD S. , NUSTAD K. , REITH A. , UGELSTAD J.
Published: June 2006

The interaction between polymer particles with an immune ligand on their surfaces and normal (hepatocytes) and malignant (Raji) cells with a corresponding membrane receptor was investigated by scanning and transmission electron microscopy. The influence of time, temperature, and anti‐metabolites on (his process was studied. The results indicate that when particles with an immunoligand are added to cells with a corresponding membrane receptor, three successive steps are initiated. the first two of which are not dependent on the temperature or the metabolic state of the cells. The first step in this process is the binding of particles to the cell surface through the immunoligand on the particle and the receptor on the cell surface. The initial binding formed by immunoligand and corresponding receptor is followed by a stronger interaction between cell and particle. This second step seems to be governed by a successive antibody‐antigen interaction: the zipper mechanism. A third step in the particle‐to‐cell interaction is characterized by movements of the cellular surface, as revealed by membrane folds, pseudo‐pods, and lamellae extending from the surface and enveloping large parts of the particle.
Characterization of Raji Cell Binding IgG in Patients with Metastatic Breast Cancer
Authors: KRIEGER G. , VOLLING P. , KNEBA M. , BOLZ I. , NAGEL G. A.
Published: June 2006

The aim of the present study was to isolate and characterize the immune complexes detected by the Raji cell assay in metastatic breast cancer. However, the Raji cell binding material could not be separated from monomeric IgG by Sephacryl S‐300 gel chromatography in any of the 16 sera investigated. The parallel elution profile of monomeric IgG and the Raji cell binding activity suggested that anti‐Raji cell antibodies, rather than immune complexes of low molecular weight, were present in these sera. This was further substantiated by pepsin digestion of the Raji cell binding IgG fractions. The binding of F(ab')2 fragments was quantitatively comparable to the binding of undigested IgG, and the bound F(ab')2 fragments could be visualized by immunofluorescence at the cell membrane. The presence of antibodies against Raji cells was further confirmed by the complement‐dependent cytotoxicity assay. These antibodies did not react with untransformed lymphocytes and there was no correlation with anti‐Epstein‐Barr virus antibodies. The incidence of cytotoxic anti‐Raji cell antibodies in breast cancer was 12% compared to 20% in malignant lymphoma, to 0% in normals and patients with degenerative cardiovascular diseases, and to 5% in patients with auto‐immune diseases.
T‐Cell Development during Ageing in Congenitally Athymic (Nude) Rats
Authors: VAESSEN L. M. B. , BROEKHUIZEN R. , ROZING J. , VOS J. G. , SCHUURMAN H.‐J.
Published: June 2006

We studied the thymus‐dependent immune system in congenitally athymic (nude) rats of the WAG (RT‐1a) strain, at the ages of 2, 3, 5, 9 and 17 months. This included the histology of spleen, lymph nodes, and Peyer's patches, immunohistochemistry on tissue sections, and immunocytology on cell suspensions using a panel of monoclonal antibodies to T‐lymphocyte subpopulations. in vitro mitogen responsiveness, and in vivo responsiveness to ovalbumin immunization. The results were compared with those in euthymic immunocompetent litter‐mates of the same age. The cell marker analysis, especially in nude animals, was hampered by the staining of non‐T cells by some of the antibodies (staining of putative macrophages, natural killer cells, cells of H‐lymphocyte lineage, and of myeloid lineage I. Despite this, with age an increase in cells with these markers was observed; for instance, in spleen suspensions, cells labelled by the pan‐I reagent MRCOX‐19 increased from 5% at 2 months to 19% at 17 months (value in euthymic animals 34%). In the other assays too a gradual increase of T‐cell reactivity with age was observed, from almost absent in 2‐ and 3‐month‐old nude rats to values in 17‐month‐old animals of about hall the level round in euthymic rats. However, none of the nude animals responded to ovalbumin immunization. These results indicate that in nude animals processes occur which compensate for the absent intrathymic T‐cell generation. This applies in particular to changes in T‐cell phenotype and mitogen responsiveness, but not to antigen responsiveness.
Immune Responses during Human Schistosomiasis Mansoni
Authors: GARCIA A. A. , da SILVA A. L. , de OLIVEIRA L. R. , KATZ N. , GAZZINELLI G. , COLLEY D. G.
Published: June 2006

Splenocytes from 25 patients with severe hepatosplenic schistosomiasis mansoni were obtained after therapeutic splenectomy, Spleen cells were phenotyped and analysed for responsiveness to mitogens or heterogeneous schistosome‐derived antigenic preparations (eggs. SEA; adult worms, SWAP; cercariae. CERC) in blastogenesis assays and lymphokine production systems, and were compared with peripheral blood mononuclear cells (PBMN). Splenic lymphocytes were 55% T lymphocytes (sheep erythrocyte rosette‐positive) and 37% surface immunoglobulin‐positive B lymphocytes. The mean T4+:T8+ ratio of these splenocytes was 1.0. Phytohaemagglutinin stimulated spleen cell production of the lymphokine mitogenic factor, but exposure to SEA or SWAP did not. Spleen cell and PBMN blastogenic responses to SEA and SWAP were sometimes, but not always in accord. Removal of plastic adherent cells allowed the non‐adherent spleen cells of 30–40% of the patients to respond substantially more vigorously to SEA, SWAP and CERC. Spleen cells from a subgroup or 20–30% of the patients failed to respond to the schistosomal antigens regardless of removal of adherent cells. Spleen cell responses to gram‐negative lipopolysaccharide peaked on day 5 or 6 of culture, and were augmented by adherent cell removal. Pokeweed mitogen‐stimulaled responses were optimal on day 5 of culture. Spleen cells from most severe, hepatosplenic schistosomiasis mansoni patients do not respond well to schistosomal antigens or B‐cell mitogens. The splenic responses of many of these patients were elevated by the removal of adherent spleen cells.
Generation of Agammaglobulinaemic Mice by Prenatal and Postnatal Exposure to Polyclonal or Monoclonal Anti‐IgM Antibodies
Authors: CERNY A. , HEUSSER Ch. , SUTTER S. , HUEGIN A. W. , BAZIN H. , HENGARTNER H. , ZINKERNAGEL R. M.
Published: June 2006

Improved experimental conditions are described for the treatment of mice with anti‐IgM antibody, which subsequently lead to B‐cell deficiency and agammaglobulinaemia. Antibody transmitted via maternal milk alone was found to be more efficient in inducing suppression of serum immunoglobulin isotypes than prenatal transmission or postnatal intraperitoneal injections alone. However, the combined treatment by all three routes of exposure to anti‐IgM resulted in total B‐cell suppression associated with undetectable levels of all serum immunoglobulin isotypes. Furthermore, suppression of B‐cell generation was also achieved with a rat monoclonal μ‐specific antibody. The possibility of generating agammaglobulinaemic mice may be useful for investigating the influence of B cells on the generation of T‐cell reactivities, and for analysing the effects of monoclonal antibodies in the absence of interfering serum immunoglobulin.
Role of Serine Proteases of Schistosoma mansoni in the Regulation of IgE Synthesis
Authors: VERWAERDE C. , AURIAULT C. , DAMONNEVILLE M. , NEYRINCK J.L. , VENDEVILLE C. , CAPRON A.
Published: June 2006

The regulation of the IgE response by schistosomula‐released products (SRP) was studied either in vitro with rat and human cell cultures or in vivo by injection into rats of SRP with an unrelated allergen at primary or secondary immunisation. The results obtained in vitro showed that non‐dialysable factors present in SRP potentiate the IgE synthesis by rat and human cells. This enhancing effect was supported by molecules with serine protease activities. On the other hand, the inhibition or depletion of SRP in serine proteases induced a weak synthesis of IgM by rat cells in vitro. The injection of SRP into rats on day 0 with an unrelated allergen led to a potentiation of total IgE production, but an inhibition of specific IgE response. In contrast, a marked elevation or specific IgE response was obtained when SRP was injected upon secondary immunization. Serine proteases of SRP were partly responsible for this potentiative effect.
Functional Competence of Dendritic Cells of Ageing C57BL/6 Mice
Authors: KOMATSUBARA S. , CINADER B. , MURAMATSU S.
Published: June 2006

Highly poly morphic age‐dependent differences in progression of T and B cells have been found in various strains of inbred mice. In the immune system of C57BL/6 mice these occur first in B cells and, later in life, in T cells. In this paper, we have examined the progression in the functional capacity of C57BL/6 dendritic cells (DC). Age‐dependent changes were found in the syngeneic mixed leucocyte reaction, in which the stimulatory capacity of DC increased with age. This increase was independent of the age of the donor of the serum that was added to the culture, and occurred when young donors provided the T cells. Age‐dependent changes in DC properties were not detected in terms of direct plaque‐forming response, response to con‐canavalin A, or in the allogeneic mixed leucocyte reaction.
Proportions of IgG Subclasses in Antibodies Formed during Vaccination with Hepatitis B Surface Antigen
Authors: MATTILA P. S. , PELKONEN J. , MÄKELÄ O.
Published: June 2006

Nineteen healthy adults were given three injections of hepatitis B vaccine (days 0, 30. and 180), and serum samples were obtained on days 0, 21. 51. and 201. A fourfold or greater increase of IgG antibodies within 201 days was detected in 18 individuals (geometrical mean increase 26‐fold). but final IgG antibody concentrations (0.3‐3μgml) were low Some vaccinees had almost reached peak concentration on day 21. in others a great increase was observed between days 21 and 51. and in a few individuals between days 51 and 201. IgG1 was the predominant subclass in the antibodies of most individuals Ninety‐five percent of all IgG antibodies were IgG1 in early samples of early responders. In late samples of all vaccinees the average proportion of IgG1 antibodies was 78% The proportion of IgG2 antibodies was slightly higher in late samples (average 12%) than in the early samples of the early responders. (average less than 6%, P<0.05). Considerable amounts of IgG3 and IgG4 antibodies were found in one vaccinee, but on the average the proportion of IgG3 or IgG4 antibodies was low (less than 6%).
Detection of Tumour Necrosis Factor‐Like Cytotoxicity in Serum from Patients With Septicaemia but Not from Untreated Cancer Patients
Authors: WAAGE A. , ESPEVIK T. , LAMVIK J.
Published: June 2006

With a bioassay sensitive for recombinant tumour necrosis factor at a concentration of 10‐4 ng/ml, we were able to detect tumour necrosis factor‐like cytotoxicity in serum from three patients in the initial phase of septicaemia. The cytotoxic activity corresponded in a concentration of recombinant tumour necrosis factor of 2x10‐2‐3x10‐3 ng/ml serum. The concentration in cerebrospinal fluid was estimated lo be three limes higher than in serum. In 23 untreated patients with malignant disease of varying origin and stage, including two patients with severe cachexia, we were not able to detect tumour necrosis factor‐like cytotoxicity.
The Distribution of Class‐Specific Rheumatoid Factors Is Similar in Rheumatoid and Pre‐Illness Sera
Authors: TUOMI T. , PALOSUO T. , AHO K.
Published: June 2006

The occurrence of IgM, IgG, and IgA class rheumatoid factors (RF) was compared by means of ELISA in 21 sera from patients with established rheumatoid arthritis (RA) and in 22 pre‐illness sera from subjects who 4 months to 5 years later developed seropositive rheumatoid arthritis. The positive cases were virtually confined to three groups of about equal size: positive for all three RF classes, positive for IgM‐RF and IgG‐RF, and positive for IgM‐RF only. Thus, the distribution of the RF isotypes was similar in rheumatoid and pre‐rheumatoid sera.
Kinetic Analysis of Interleukin 2 (IL‐2) Production and Expression of IL‐2 Receptors by Uraemic and Normal Lymphocytes
Authors: LANGHOFF E. , HOFMANN B. , ØDUM N. , LADEFOGED J. , PLATZ P. , RYDER L. P. , SVEJGAARD A.
Published: June 2006

The in vitro immune response of lymphocytes from uraemic patients was studied by comparing the in vitro kinetics of interleukin 2 (IL‐2) production, the mitogen‐induced proliferative response, and the expression of IL‐2 receptors by T lymphocytes. The IL‐2 production in 26 uraemic cell cultures decreased significantly after 48 h of stimulation with mitogen compared with that of 24 control cultures. The lymphocyte responses to phytohaemagglutinin (PHA). concanavalin A (Con A), and pokeweed mitogen (PWM) increased linearly with time, but the responses of the uraemic cells cultures were significantly lower than those of the control cultures. The relative numbers of cells double‐stained for both Tac (IL‐2 receptor)/HLA‐DR or Tac/Leu 2 were significantly increased in the uraemic cultures as compared with the control cultures at 48 and 72 h. A similar, but not significant, trend was also demonstrated for uraemic cells positive for Tae/Leu 3. These findings were also seen in uraemic lymphocyte cultures supplemented with exogenous IL‐2. Thus, the IL‐2 production of uraemic lymphocytes seems to be exhausted more rapidly than that of normal lymphocytes, and there is no evidence that the poor proliferative response of uraemic lymphocytes is due to a decreased relative number of cells positive for IL‐2 receptors.
Stages in LAV/HTLV‐III Lymphadenitis
Authors: PALLESHN G. , GERSTOFT J. , MATHIESEN L.
Published: June 2006

Lymph nodes from 40 homosexuals with persistent, generalized lymphadenopathy were studied for histological and immunohistological changes and classified into histological stages based on the progressive destruction of lymph node follicles in association with progression of the disease. Three patterns were recognized: stage I was characterized by follicular hyperplasia, mantle zone depletion, and follicular fragmentation in the absence of vasculities, stage II by signs of follicular involution, and stage III by depletion of follicles and dendritic reticulum cells with development of diffuse pattern. The T zone was gradually depleted of CD4‐positive lymphocytes, but on further progression. lymphocytic depletion (which also involved the CD8‐positive cells) and fibrosis prevailed. The 40 lymph nodes from homosexuals were classified as stage I in 18 cases, stage II in 11, and stage III in 10. One case did not fulfil our histological criteria for LAV/HTLV‐III lymphadenitis, although this patient was seropositive. Convincing correlation was found between histological stages and clinical and laboratory data. The triad of follicular hyperplasia. mantle zone depletion, and follicular fragmentation, in the absence of vasculities, appears pathognomonic for the early disease. The diffuse pattern, however, may be seen in different disease entities.
Analysis of Monoclonal Antibodies to Mycobacterium leprae by Crossed Immunoelectrophoresis
Authors: HARBOE M. , IVANYI J.
Published: June 2006

Monoclonal antibodies to Mycobacterium leprae were characterized in crossed immunoelectrophoresis and showed markedly different patterns of reactivity with M. leprae lines 2, 7 and 11 respectively. Line 7 corresponds to a cell wall‐associated macromolecular complex containing lipid, polysaccharide, and two distinct 36 K and 65 K proteins.
The Effect of Tunicamycin on Target Cell Susceptibility to Natural Killer Cell Cytotoxicity
Authors: NOSE M. , GIDLUND M. , HOSEIN Z. , AXBERG I. , WIGZELL H. , YOGEESWARAN G.
Published: June 2006

Several sets of data indicate the possibility that carbohydrate moieties on the target cell are important structures in natural killer (NK) cell‐mediated lysis. Strkining changes in the NK susceptibility of targets can be induced in several systems involving in vitro differentiation of tumour cell lines. The effect on target cells of the glycosylation inhibitor tunicamycin, which acts by blocking the dolichol‐dependent asparagine‐linked glvcosvlation pathway was investigated. Using several different tumour cell lines we can conclude that; (a) asparagine‐linked carbohydrate chains do not contribute directly to NK susceptibility, (b) induced differentiation may or may not be linked with a change in NK susceptibility, and (c) secondary changes caused by tunicamycin treatment may lead to alterations in the gangliosides, a finding that is positvely correlated with decreased NK susceptibility.
Antigenic Variants of Human IgG Subclasses
Authors: OXELIUS V. ‐A.
Published: June 2006

A comparison of murine monoclonal IgG subclass antisera with rabbit polyclonal IgG subclass antisera restriction of the monoclonal antisera and further antigenic heterogeneity within the IgG subclasses of at least two antigenic variants of IgG1, two of IgG2, two of IgG3 and a third subtype of IgG4.
Frequency and Functional Characterization of Specific T‐Helper Cells Infiltrating Rat Kidney Allografts during Acute Rejection
Authors: MANCA F. , FERRY B. , JAAKKOLA M. , HALTTUNEN J. , HORSMANHEIMO L. , HÄYRY P.
Published: June 2006

T‐helper cells (ThC) play an important role in the induction of both cytotoxic T‐cell responses and B‐cell responses against the grafted organ. Furthermore, ThC alone are capable of causing graft rejection in T cell‐deprived mice and rats. In view of these observations we found it important to analyse the frequency and functions of donor‐specific ThC in the allograt and in the recipient lymphoid system during the course of acute renal allograft rejection. A limiting dilution assay was developed which, due to the absence of exogenous interleukin 2 (IL‐2) and the low numbers of stimulator cells used, appears to be highly selective for the proliferation of specific ThC. Kidney transplants were performed from LBN (RT1n) to congenic Lewis (RT11) sirain differing in major histocompatibilily complex (MHC) only. The inflammatory (white) cells were recovered from the graft, and blood and recipient spleen and the frequency of RT1n‐ responding ThC were determined at different times after transplantation. In the kidney graft itself, the frequency of ThC responding to RT1n MHC antigens was 1:3000 on day 2 and increased to l:670–1320 at the peak of inflammation. In the spleen, the frequency increased from l:l000 on day 0 to 1:200 on day 8, and remained high even after the graft was rejected. In the blood, the frequency stayed at the 1:400–1:800 level, and increased to 1:200 only after the graft had been completely destroyed. Individual ThC clones deriving from limited dilution assays of kidney and spleen cells were recovered and expanded with irradiated donor cells without IL‐2 and finally with exogenous IL‐2 only. All clones showed the T‐helper (W3/25) phenolype, seven out of eight tested clones showed a specific anamnestic response lo RT1n alloantigens and no response to RT11 or RT1a in a secondary MLC, 12 out of 12 clones produced IL‐2 and 11 out of 11 clones produced gamma interferon upon re‐stimulation with relevant allogeneic cells, and eight out of ten clones collaborated with syngeneic B cells for Ig synthesis, indicating that they were indeed derived from specific ThC and/or from their precursors. Taken together, the results demonstrate that specific ThC and/or their precursors represent only a very small minority in the graft‐infiltrating inflammatory population. This makes it most unlikely that ihe ThC themselves are responsible for graft destruction; the results indicate rather that a major role of ThC in situ may be instruction of immunologically specific and nonspecific components of inflammation.
Formation of the Membrane Attack Complex of Complement (MAC) on Erythrocytes from Monocyte‐Produced Terminal Complement Components
Authors: HETLAND G. , JOHNSON E. , ESKELAND T.
Published: June 2006

By using antibodies against C5, C6, C7, C8, and C9, we found that terminal complement components were deposited on IgM‐coated sheep erythrocytes (EIgM) kept in scrum‐free endotoxin‐stimulated monocyte cultures for 24 or 48 h. Monoclonal antibodies revealed C9 neoantigens on the EIgM. There was no specific binding of an anti‐S protein antibody, which reacts with the SC5b‐9 complex, to the EIgM. Controls were native sheep erythroeytes (E) treated similarly which, in contrast to EIgM, do not activate the classical pathway of complement. Cycloheximide (l.O μg/ml) in the cell cultures resulted in no specific binding of the anti‐C9 antibodies to EIgM. A fraction of the EIgM was lysed during incubation with the monoeytes. We conclude that the monoeytes secrete C5, C6, C7, C8, and C9, which form the membrane attack complex of complement (C5b‐9) on the EIgM.
Immunological Properties of Antigen 60 of BCG Induction of Humoral and Cellular Immune Reactions
Authors: COCITO C. , BAELDEN M. C. , BENOIT Ch.
Published: June 2006

Aniigen 60(A60). a member of the thermostable macromolcular antigen family (TMA) and main component of old tuberculin and purified protein derivative (PPD), has been purified from the cytoplasm of Mycohacterium bovis BCG; its structure and metabolism have already been described. In the present paper, the action of A60 on humoral immunity has been analysed by an ELISA type immunoassay, and that on cellular immunity by the mouse footpad swelling test. Injection of very low A60 doses into unprimed mice produced an undetectable level of anti‐A60 antibodies; the effect or a booster inoculation was not appreciable in the absence of incomplete Freund's adjuvant, but was evident when the latter was added. Higher doses of the antigen produced an appreciable primary response, and a sharp and long‐lasting secondary response, which had a 10‐fold higher intensity in the presence of incomplete adjuvant. No detectable delayed hypersensitivity reactions were observed in unprimed mice after footpad injeciion of A60, whereas clear responses were elicited in primed mice. This effect was more pronouneed when the footpad was injected after a secondary response than after a primary response, and it was invariably magnified by incomplete adjuvant. It Is concluded Ihat A60 is a powerful immunogen. which is able to induce primary and secondary responses;md delayed hypersensitivity reactions, effects that are adjuvant‐modulated and develop concurrently.
Wheat Germ Agglutinin is Mitogenic, Non‐Mitogenic and Anti‐Mitogenic for Human Lymphocytes
Authors: KILPATRICK D. C. , McCURRACH P. M.
Published: June 2006

Wheat germ agglutinin (WGA) was found to stimulate DNA synthesis in human peripheral blood mononuclear cells at relatively low concentrations and to inhibit DNA synthesis at higher concentrations. Both actions were inhibited by oligomers of N‐acetyl‐D‐glucosamine. Significant mitogenic activity was dependent on the use of human (as opposed to fetal calf) serum to supplement the culture medium. Purified T cells responded to WGA very weakly and the incorporation of thymidine into non‐T cells in response to WGA was less than the lectin‐free control. The full ability of T cells to respond to WGA was restored by the addition of monocytes, but not by any other blood cells. Interleukin 2 partially restored the ability of T cells to respond to WGA;interleukin 1 was less effective. WGA displayed a strong synergistic action with the tumour promoter, 12‐O‐tetradecanoyl‐13‐acetate (TPA), in stimulating DNA synthesis in separated T (but not non‐T) cell fractions, and in unfractioned mononuclear cells. These results reconcile most of the conflicting reports in the literature concerning the interaction of WGA with human lymphocytes.
Evidence of an Activated T‐Cell System with Augmented Turnover of Interleukin 2 in Rheumatoid Arthritis
Authors: WAALEN K. , FøRRE ø. , LINKER‐ISRAELI M. , THOEN J.
Published: June 2006

The stage of activation of synovial inflammatory T cells was studied in rheumatoid arthritis and compared with that of normal T cells stimulated in vitro by dendritic cells. Increased numbers of rheumatoid inflammatory T cells expressed activation antigens such as HLA‐DR, interleukin 2 receptors (Tac), and transferrin receptors. These T cells also had high spontaneous proliferation. Peripheral blood T cells stimulated by autologous dendritic cells showed a similar expression of activation antigens and had high proliferation. Resting T cells and T cells cocultured with monocytes expressed much less activation markers. Exogenous interleukin 2 significantly increased the proliferation of the various T cells. All responses were inhibited by an antibody which blocks the interleukin 2 receptor. Supernatants from mitogen‐stimulated rheumatoid inflammatory mononuclear cells and mitogen‐stimulated T cells preactivated by dendritic cells usually contained less interleukin 2 than supernatants from mitogen‐stimulated normal T cells. Furthermore, the various in vivo and in vitro‐activated Tac‐positive T cells, in contrast to nonactivated Tac‐negative T cells from peripheral blood of patients and controls, significantly absorbed interleukin 2. The results indicate a turnover of interleukin 2 in rheuniatoid inflammatory T cells comparable to that of T cells activated by dendritic cells.
Long‐Term Cholera Antitoxin Memory in the Gut can be Triggered to Antibody Formation Associated with Protection within Hours of an Oral Challenge Immunization
Authors: LYCKE N. , HOLMGREN J.
Published: June 2006

A local mucosal immunoiogical memory that could be efficiently triggered to protective antibody formation on renewed antigen exposure might account for the several‐year long protection against reinfection and disease seen in individuals after cholera disease. The duration and other functional aspects of gut mucosal immunological memory to the cholera toxin (CT). which is the key pathogenic factor in cholera, were examined in mice. Six months or even 2 years after an initial series of oral immunizations with CT a single repeat oral exposure to CT in submicrogram amounts evoked a brisk IgA antitoxin response in the lamina propria. A three‐fold increase in IgA antitoxin‐producing cells (SFC) was evident within 16 h, with a further rise in SFC numbers over the next several days. The anamnestic gut mucosal IgA antitoxin response was associated with a substantial increase in protection against challenge of intestinal loops with CT. The rapid increase in IgA antitoxin SFC in the gut is believed to reflect memory cells dispersed in the gut mucosa which can be rapidly triggered into antitoxin formation by antigen encounter in vivo and such cells could clearly be responsible for the longterm immunity seen after cholera disease or oral vaccination.
Protection against Cholera Toxin after Oral Immunization is Thymus‐Dependent and Associated with Intestinal Production of Neutralizing IgA Antitoxin
Authors: LYCKE N. , ERIKSEN L. , HOLMGREN J.
Published: June 2006

We studied whether gut mucosal IgA antitoxin production as well as the acquired protection against cholera toxin (CT) after oral immunization with CT are both thymus‐dependent immune manifestations. In contrast to normal BALB/c mice, nude, athymic mice did not respond to oral immunizations with CT with either IgA antitoxin‐producing cells (SFC) in the lamina propria or protection against challenge with CT in ligated intestinal loops. However, when nude mice were first reconstituted by grafting of syngeneic thymus glands, both IgA antitoxin SFC in the lamina propria and protection were stimulated by oral immunizations with CT and the responses were of similar magnitude to those of normal mice after immunizations. During in vitro culture, isolated lamina propria lymphocytes from immunized but not from control mice concomitantly and proportionally produced IgA antitoxin and CT‐neutralizing activity. We conclude that intestinal antitoxin formation and protection against toxin challenge after oral immunization with CT are both critically thymus‐dependent and therefore likely to be under T‐cell control.
The Gene for the Leu‐M2 (Mac‐120) Antigen is Located on Human Chromosome 11
Authors: SUOMALAINEN‐NEVANLINNA H. A. , SCHRÖDER J. , ANDERSSON L. C.
Published: June 2006

Leu‐M2 (Mac‐120) antigen is a cell‐surface marker present on a subpopulation of human blood monocytes and platelets. Its expression on monocytes and macrophages is necessary for their ability to present antigen to T lymphocytes. Only Leu‐M2+ cells display the human la‐like determinant, HLA‐DS, which seems important in antigen presentation and stimulation in the autologous MLR. Expression of the Leu‐M2/Mac‐120 antigen was studied by indirect immunofluorescence analysis and flow cytofluorometry in mouse‐human somatic cell hybrids segregating human chromosomes. In the mouse‐human hybrid clones, expression of the Leu‐M2/Mac‐120 antigen was dependent on the presence of human chromosome 11. This was verified by fluorescence‐activated cell sorting of heterogeneous hybrid cell populations into antigenpositive and ‐negative fractions. The Leu‐M2 antigen co‐segregated with chromosome 11. The gene controlling the expression of the Leu‐M2/Mac‐120 antigen thus is assigned to human chromosome 11.
Purification of an IgM Antibody Response‐Enhancing Factor to the Alpha 1–6 Epitope of Native Dextran from Serum and Supernatants from T‐Cell Hybridomas
Authors: PASANEN V. J. , LEACH R. , MOLLER G.
Published: June 2006

We have found that some hybrids produced by fusing AKR thymoma BW 5147 wiih spleen cells from Dx‐hyperimmunized CBA mice produce factors that affect the primary CBA IgM PFC response. By the use of HPLC‐DEAE chromatography, sometimes combined with affinity chromatography. we have purified from hyperimmunc anii‐Dx serum, from ascitesgrowing T hybrids, and from normal CBA serum a family of factors that specifically enhance the anti‐Dx IgM response. They show specificity for either Dx or anti‐Dx, they may contain an Iak determinant, and they are genetically restricted in their function. The factors from all three sources are found in ihe same fractions after separation and move in fractions without Ig contamination. The affinity‐purified factor was eluled at an NaCl concentration of 0.15–0.20 m. On the basis of these characteristics we believe that we have separalcd a class of T cell‐derived helper factors from serum. The concentration of the factor in serum increases after hyperimmunizaiion with Dx. As a specificity control we show that the same sera contain a natural anti‐SRBC IgM response‐enhancing factor. It co‐separates with the anti‐Dx‐enhancing factor, but they can be distinguished by their preferential binding to their own antigens.
The Origin of the Immune System
Authors: MATSUNAGA T. , MORI N.
Published: June 2006

It has been suggested that the neural cell adhesion molecules (N‐CAM) are members of the Ig superfamily. We have exaniained structural homology between the N‐CAM and other superfamily molecules in detail. The Ig‐like domains of N‐CAM have overall sizes similar to ihoseof Ig constant domains or major histocompalibitity complex (MHC) domains. Significant sequence homology was found between the N‐CAM Ig‐like domains and several other Ig superfamily members, such as T4. TH. IgA transport receptor, Ig VH. Ig Cμ4, and others. A comparison of the second domain of T4 (255 amino acids) wiih ihe central domain of N‐CAM (302 amino acids) has demonstrated a significant sequence homology. Furthermore, a cell adhesion protein, cs‐A, of slime mould, Dictyostelium, also seems to be related to N‐CAM. It appears that the Ig superfamily of the immune system originated from more ubiquitous ceil recognition moleeules whieh are eommon in all metazoan species.
Reactive Oxygen Species Selectively Deplete Normal T Lymphocytes Via a Hydroxyl Radical Dependent Mechanism
Authors: ALLAN I. M. , LUNEC J. , SALMON M. , BACON P. A.
Published: June 2006

Chronic inflammatory synovitis is characterized by both lymphocytic infiltrates and persistent polymorph exudates. Activated polymorphs release reactive oxygen species (ROS) during inflammation, but the contribution that these make to the lymphocyte abnormalities associated with RA has been little studied. We therefore investigated the cytotoxic effects of the reactive oxygen speices on human peripheral blood mononuclear cells (PBMC). PBMC were exposed to RPMI 1640 medium previously irradiated for up lo 60 min. Consistent dose‐dependent killing was observed at 24 h. Antioxidant studies indicated that H2O2 was the effective species. Catalase, which specifically degrades H2O2, gave almost total protection against cell death, while superoxide dismutase (SOD), thiourea, and mannitol were largely ineffective. Addition of exogenous H2O2 caused an identical pattern of cell death to that observed with irradiated medium. PBMC cultures supplemented with desferrioxamine (a ferric iron chelator) also gave significant protection, suggesting that H2O2 mediated its effects via OH* radicals, Analysis of lymphocyte subpopulalions showed that ROS caused a selective depletion, depending on the level of H2O2 present. Low levels induced a speciftc loss of CD8+ cells, while higher concentrations caused significant loss of CD4+ T cells as well. slg+ B cells were unaffected at either concentration. This selective lymphotoxic effect of ROS may be of considerable importance in the pathogenesis of autoimmune inflammatory disease.
Matching of Host Genotype and Serotypes of Coxsackie B Virus in the Development of Juvenile Diabetes
Authors: FOHLMAN J. , BÖHME J. , RASK L. , FRISK G. , DIDERHOLM H. , FRIMAN G. , TUVEMO T.
Published: June 2006

Thirty‐six consecutive paediatric patients (0‐16 years old) with recently contracted juvenile diabetes (IDDM) during 1982‐84 were included in the study. Sera were assayed for recent or current Coxsackie B virus (CBV) infection using a specific and sensitive IgM RIA. Eighteen patients (50%) had IgM against CBV 1‐5. The patients were also assayed for restriction fragment length polymorphism (RFLP) patterns with DNA probes coding for HLA‐DR and DQ beta chains. The CBV‐positive patients (n=18) had either RFLP patterns associated with HLA‐DR 3 or 4 or HLA‐DQ patterns III or IV beta. Two of the CBV negative patients had neither HLA‐DR 3 nor DR 4 and four of them had neither DQ patterns III nor IV. Eleven out of 18 CBV‐positive patients had HLA‐DQ III and DR 3 (61%) versus 5 out of 18 (28%) of the CBV‐negative patients. All 11 patients with serology positive for CBV 2,3, and 5 had HLA‐DR 4 and DQ IV patterns. This was significantly (P<0.01) different from all five CBV 4‐positive patients, who in contrast all had HLA‐DR 3 or HLA‐DQ III patterns. CBV 1‐positive patients (n=2) all had HLA‐DR 3, 4, and HLA‐DQ III, IV patterns. Thus CBV 4 seems to be significantly associated with a different host genetic constitution from at any rate CBV 2,3, and 5, and possibly CBV 1.
Induction of Salivary Antibody Responses in Rats after Immunization in Peyer's Patches
Authors: DAHLGREN U. I. H
Published: June 2006

We examined salivary, milk, and serum anlibody levels after immunization in the Peyer's patches (Pp) of rats with horse spleen ferritin. Priming ofthe Pp one day after parturition led to the appearance of IgG, bit not IgA or IgM, anti firritin antibodies in saliva 9 days later. IgG and IgM antibodies were detected both in milk and in serum, whereas IgA antihodies could only be demonstrated in milk. During a second lactation period the salivary antibodies had vanished but IgG antibodies could still be detected in milk and serum. During a third location period, when the rats were immunized in the Pp a second time, not only IgG but also IgA antiferritin antibodies appeared in the saliva. Salivary IgG antibody levels and milk IgG, IgM, and IgA antibody levels were higher than those observed after primary immunization in the Pp. The IgG antibody activity in the saliva was positively correlated to the serum IgG antibody activity. It is concluded that salivary IgA antibody responses can be induced by immunization in the Pp. The results of this study suggests that IgA antibodies delected in saliva are produced locally by cells that have migrated from the intestinal lymphoil tissue to the salivary glands.
Activation of Resting, Pure CD4+, and CD8+ Cells via CD3
Authors: HALVORSEN R. , GAUDERNACK G. , LEIVESTAD T. , VARTDAL F. , THORSBY E.
Published: June 2006

We studied the requirements for secondary activation signals in pure CD4+ and CD8+ T cells after stimulation with anti‐CD3 antibodies. Stimuluion or CD4+ or CD8+ cells with anti‐CD3 monoclonal antibodies (MoAb) bound to polyptyene monosized particles never resulted in a proliferative response. However, DNA synthtsis was observed when recombinant interietikin 2 (IL‐2) or other secondary signals, such as those provided by phorbol myristate acetate (PMA) or autologous accessory cells (AC), were also added. These secondary signals were not in themselves capable of inducing DNA synthesis in the absence of particle‐bound anti‐CD3. We also found that the signals provided by AC may be dependent on the activation state of these cells. Thus, the effects of accessory cells were enhanceed by a factor present in fetal calf serum (FCS), must likely endotoxin or lipopolysaccharide (LPS), which alone, however, were not able to activate T cells, even in the presence of particle‐bound anti‐CD3. Recombinant lL‐1 over a broad dose range was unable to replace PMA or activated AC after slimulation with particle‐bound anti‐CD3. Purified CD4+ and CDS+ T cells behaved idenlically in all the experiments, indicating that the basic mechanisms for activation in the two T‐cell subsets are idenlical.