bluuebunny/crossref_metadata_2025 huggingface dataset:
Intrathecal Immune Responses in Mumps Meningitis Patients
Authors: REUNANEN M. , SALONEN R. , SALMI A.
Published: June 2006

The in vitro 3H‐thymidine incorporation of peripheral blood (PB) and cerebrospinal fluid (CSF) lymphocytes from 11 mumps meningitis patients was studied after stimulation with non‐specific mitogens and microbial antigens. Although corresponding viral antibodies were always found in the CSF of seropositive patients by a sensitive radioimmunoassay, their intrathecal synthesis was directed mainly against mumps virus. Most of the patients had PB lymphocytes that reacted on stimulation with phytohaemagglutinin (PHA) or pokeweed mitogen (PWM), but a smaller number of them had reactive CSF cells. Only four patients showed stronger responses to PHA in the CSF than in PB. Most patients had mumps‐reactive lymphocytes in PB but only two of them in the CSF. In contrast, these patients more often had increased CSF cell reactivity when tested with purified protein derivative, measles, and herpes simplex virus antigens. The results could not be explained by a damaged blood—brain barrier alone but may reflect the immunological status of the brain compartment in these patients.
Role of the Popliteal Lymph Node in Infection with Mycobacterium marinum of the Hind Footpad of the Mouse, and Source of the Cells that Characterize the Process
Authors: MOR N. , LUTSKY I. , LEVY L.
Published: June 2006

We examined the possibility that the popliteal lymph node serves as the source of the lymphocytes that, together with macrophages, characterize Ihc lesion produced by infection with Mycobacterium marinum in the hind footpad of Ihc mouse. Naive mice were partially protected against challenge with M. marinum in the hind footpad by intravenous infusion of lymphoc) its harvested from the popliteal nodes of donor mice infected with M. marinum 7 days earlier. Lymphocytes harvested from the popliteal nodes of infected donors, labelled in vitro with 3H‐uridine, and infused intravenously into naive mice that were immediately challenged in the hind footpads with M. marinum, localized in the popliteal nodes of the recipient mice but nol in the footpad lesions. Lymphocytes harvested from the spleens of naive donors and labelled in vitro appeared to home to the popliteal nude draining the M. marinum‐infected footpad. Thus, the primary rule of the popliteal lymph node appeared to be passive trapping of the lymphocytes brought to it by the circulairon or afferent lymphatics. We then tried lo locate the sources of bolh lymphocytes and macrophages that characterize the lesion. Temporary occlusion of the abdominal aorta prevented labelling by intravenously infused 3H‐thymidine (3H‐TdR) of the mononuclear cells of both footpad lesion and popliteal node. Temporary occlusion of the left common iliac artery during 3H‐TdR infusion prevented immediate labelling on the ipsilateral side. After 24 and 48 h, however, small numbers of labelled lymphocyles were found in the left hind footpad lesion. Amputation of the right leg at the hip joint, but run right poplitial lymphadenectomy, performed immediately after re‐estublishment of patency of the left common iliac artery, prevented the late influx of labelled lymphocytes into the lesion of the left hind footpad. Thus, the chief source of both the lymphocytes and the macrophages of the footpad lesion appeared lo be the lesion ilself.
A Synergistic Polyclonal Response to Dextran Sulphate and Lipopolysaccharide
Authors: BERGSTEDT‐LINDQVIST S. , FERNANDEZ C. , SEVERINSON E.
Published: June 2006

The effects of the simultaneous addition of lipopolysaccharide (LPS) and dextran sulphate (DxS) were studied in a low‐cell density culture system, which enables exlensivc cell proliferation and high immuoglobulin secretion. Using these two milogens, a synergistic‐response was observed, with regard to both cell division und IgM secretion. However, only a very low IgG production could be detected. This was caused by the extensive cell proliferation. leading to suboptimal culture conditions. Thus, when the blasts, were recultured at a lower cell density or when a lower initial cell concentration was used, a high IgG response was obtained. The synergistic response induced by LPS plus DxS was independent of T cells, Furthermore, no apparent need for phagocytic cells was found. Both the LPS‐ and the LPS plus DxS‐induced activation led to a switch, preferentially to IgG2b and IgG3 secretion. This subclass pattern was not changed when the cultures were kicking functional T cells.
Inhibition of T‐Cell Activity by Cyclosporin A
Authors: ANDRUS L. , LAFFERTY K. J.
Published: June 2006

Cyclosporin A (CyA) inhibits early events in the T‐ccll response. It strongly suppresses the activation of naive T cells by IL1 or IL2 and alloantigen. CyA exerts a selective effect on activ ated T cells, It inhibits the ability of these cells to release IL2 in response to antigen or mitogen restimulation, at concentrations that have no effect on the ability of these same cell populations to respond to IL2 by proliferation. The specific effects of CyA are not limited to T cells, however, and this drug will inhibit IL1 production by lipopolysaccharide W‐stimulated PU5‐IR cells.
Isolation and Characterization of TR‐c, an Antigen of the Reiter Treponeme Precipitating with Antibodies in Syphilis
Authors: PETERSEN C. SAND , PEDERSEN N. STRANDBERG , AXELSEN N. H.
Published: June 2006

TR‐c is a Reiter treponemal antigen that cross‐reacts with an antigen in Treponema pallidum (Nichols pathogenic strain). Sera from patients with secondary syphilis contain precipitating antibodies against TR‐c. The isolation of TR‐c from a crude bacterial sonicate involves five fractionation steps: anion exchange chromatography (DE‐52 Whatman), gel filtration (Ac‐A‐22. Ultrogel), and affinity chromatography respectively on phenyl‐Sepharose CL 4B, iminodiacetic acid‐Sepharose CL 4B. and lysine‐Sepharose 4B. The purified TR‐c was enriched 320 times compared with the starting material. and the recovery was 22%, TR‐c was shown to be a protein, it did not bind to a series of lectins. and by gel filtration and polyacrylamide gel electrophoresis (PAGE) the mol, wt was determined to be in the range of 630, 000–730.000. It was found by SDS‐PAGE to be composed of identical subunits, each having a mol. wt of 48, 000. The isolated TR‐c was immunochemically pure when tested in crossed immunoelectrophoresis against polyspecific anti‐Reiter Ig. The purified TR‐c antigen was used for production of a monospecific rabbit antiserum. Monospecific rabbit anti‐TR‐c gave strong fluorescence with both the Reiter treponeme and T.pallidum.
Autologous Mixed Lymphocyte Reaction in Man
Authors: DAMLE N. K. , GUPTA S.
Published: June 2006

Human peripheral blood T lymphocytes were separated into theophylline‐resistant (TR) and ‐sensitive (TS) subpopulations. Proliferative responses of TR, TS and unfractionated T cells were studied, using irradiated autologous or allogeneic non‐T cells as stimulators. TR cells proliferated vigorously in both autologous and allogeneic mixed lymphocyte cultures (MLC). TS cells, which constitute about 20% of unfractionated T cells, exhibited poor proliferative responses to autologous and allogeneic stimulation. The magnitude of proliferation in autologous and in allogeneic MLC was found to be directly dependent on the number of TR cells in the culture. Mitomycin‐C (MMC)‐treated TR cells augmented and MMC‐treated TS cells suppressed (P<0.05) the autologous and allogeneic MLC responses of unfractionated T cells. However, the response of TS cells did not increase in autologous or allogeneic MLC when co‐cultured with TR cells. MMC‐treated TS cells, when co‐cultured with TR cells, suppressed the responses of TR cells (P<0.05). The enhancing effect of TR cells was radiation‐resistant. Suppressor influence of TS cells, in contrast, was abolished by irradiation (P<0.05). These findings demonstrate that TR cells are the responding cells in autologous MLR and augment the MLC responses of unfractionated T cells. TS cells, on the other hand, respond poorly in autologous or allogeneic MLC and suppress the response of TR cells.
Relationships between HLA‐DR‐expressing Cells and T Lymphocytes of Different Subsets in Rheumatoid Synovial Tissue
Authors: KLARESKOG L. , FORSUM U. , WIGREN A. , WIGZELL H.
Published: June 2006

An immunohistochemical double staining technique was used to study the relationships between HLA‐DR‐expressing, mostly macrophage‐like cells, and different populations of T lymphocytes in frozen sections of synovial tissue from patients with Inflammatory joint disease, particularly rheumatoid arthritis. Substantial T‐lymphocyte infiltration as measured by Leu I antibody binding was found, especially adjacent to HLA‐DR‐expressing cells near the synovial cavity and around small vessels. Most of the T cells reacted with Leu 3a ('helper/inducer'‐cell‐specific) antibodies, whereas relatively few cells were Leu 2a (cytotoxic/suppressor’ T‐cell‐specific)‐positive.
Murine Synovial Intima Contains I‐A‐, I‐E/C‐Positive Bone‐Marrow‐Derived Cells
Authors: KLARESKOG L. , FORSUM U. , WIGZELL H.
Published: June 2006

A subpopulation of murine synovial lining cells is shown to express Ia antigens coded for by both I‐A and I‐E/C loci of the major histocompatibility complex and to be derived from precursors in the bone marrow. This is demonstrated by use of biotinylated mouse monoclonal antibodies bound to frozen sections of tissue from normal and bone‐marrow‐reconstituted mice that are detected by a sensitive avidin/biotinylated peroxidase complex (‘ABC’) technique.
Detection of Antigen‐specific Cellular Immune Response by the in Vitro Production of T‐Cell Growth Factor
Authors: ILONEN J. , SALMI A.
Published: June 2006

The amount of T‐cell growlh factor (TCGF) produced in vitro in antigen‐stimulated lymphocyte cultures was measured by induction of growth in cell dependent on the factor. The method distinguishes eight herpes‐simplex‐seronegative subjects from ten seropositive ones, and these results correlated with those of the lymphocyte blast transformation (LBT) test. The LBT responses and TCGF amounts produced in cultures with purified tuberculin (PPD) were also in good correlation (r = 0.833, P<0.001). Considerable‐levels of the factor were already present 12 h after the beginning of the culture. The method offers rapid quantification of cell‐mediated immunity to specific antigens.
Unicellular or Multicellular Origin of Human T‐Lymphocyte Colonies in Soft Agar?
Authors: LINDAHL‐KIESSLING K. , KARLBERG I.
Published: June 2006

Human peripheral lymphocytes were cultivated in soft agar or agarose, containing 3H‐thymidine or bromodeoxyuridine (BrdU), which is an analogue of thymidine readily incorporated into deoxyribonucleic acid by lymphocytes. Colonies were isolated and analysed by autoradiography or stained with the fluorescence‐plus‐Giemsa technique, which identifies cells that have passed one, two, three, or more replication cycles with incorporation of BrdU. Colonies contained cells in their first and second division cycle together with later generations. The labelling index was very high. Consequently, it was concluded that, at least without special precautions being taken, lymphocyte colonies in semi‐liquid medium are likely to grow through recruitment of mobile cells as well as through divisions within the colonies. Thus, T‐cell growth in gels seems to be a matter of recognition between activated blast cells rather than monoclonal growth.
Expression of the H‐2 Antigenic Complex on Murine Haematopoietic Stem Cells
Authors: FITCHEN J. H. , HAYS E. F. , FERRONE S.
Published: June 2006

We used hybridoma‐derived monoclonal antibody to H‐2Kk antigens and xenoantibodies to β2‐microglobulin (β2) to study the expression of the H‐2 antigenic molecular complex on murine hematopoietic stem cells and the effect of long‐term culture in vitro on the expression of these antigens. Monoclonal anti‐H‐2Kk antibody produced potent complement‐dependent inhibition of pluripotent (CFU‐S), myeloid (CFU‐C), and erythroid (CFU‐E) stem cells from the bone marrow of C3H and AKR (H‐2k) mice and was without effect on stem cells from Balb/C (H‐2d) mice. Anti‐β2m xenoantibodies inhibited stem cells from all three strains. Stem cells could not be ‘rescued’ from the inhibitory effects of the antibodies by the addition of thymocytes to marrow cells after antibody treatment. Both the anti‐H‐2Kk monoclonal antibody and the anti‐β2m xenoantibodies produced potent inhibition of AKR (H‐2k) CFU‐C that had been maintained in culture for up to 6 weeks. These results indicate that murine CFU‐S, CFU‐C, and CFU‐E express H‐2 antigens and that the expression of these antigens by CFU‐C is not altered during long‐term culture.
The Generation of ‘Cytotoxic’ Macrophages in Mice during Infection with Influenza A or Sendai Virus
Authors: MAK N. K. , LEUNG K. N. , ADA G. L.
Published: June 2006

Injection of infectious but not of non‐infections influenza A virus or of infectious or non‐infectious Sendai virus intraperitoneally into mice induces the generation of plastic‐adherent cells that arc able to effect release of 51Cr from labelled virus‐infected target cells but not from labelled, uninfcctcd cells. Their activity is greatly diminished by exposure to silica or carrageenan but not by anti‐Thy I antibody and complement treatment. Similarly, the activity of the cell preparation cannot be explained by contamination with natural killer or ‘K’ cells. Thus, the effector cells were identified as macrophages and for convenience are called ‘cytotoxic macrophages’. The maximum cytotoxic activity was recovered from the peritoneal cavity 5 days after virus injection and declined thereafter. Although the effector cells are cross‐reactive in that cells activated by an influenza A strain virus lyse target cells infected with the same or other A strain viruses or with Sendai virus, there is preferential lysis of cells infected with the homologous virus. The action of the effector cells is not H‐2‐restrictcd. Preliminary experiments showed that similar effector cells can be recovered from the lungs of mice 5 days after intranasal inoculation of infectious influenza virus, so they may contribute to the host control of the disease.
Lymphokine Production in T‐Lymphocyte Subsets Defined by Active E‐Rosette Formation
Authors: ROBBINS D. S. , BELDEN E. L. , HOFFMAN P. M. , FUDENBERG H. H. , STRELKAUSKAS A. J.
Published: June 2006

Human T‐cell subsets, defined by active E rosette formation, were examined for their ability to produce leucocyte migration inhibitory factor (LIF) in response to mitogen or specific antigen. It was determined that T cells enriched for active rosette‐forming (A+) cells produced LIF in response to concanavalin A, whereas T cells depleted of active rosetteforming (A‐) cells did not. Similarly, A+ cells from a tuberculin‐sensitive donor produced LIF in response to tuberculin purified protein derivative, whereas A‐ cells from the same donor failed to produce the mediator. Thus, T‐cell production of LIF in humans appears to be restricted to those T cells capable of active rosette formation.
Receptors for the Third Complement Component on a Proportion of Large Granular Lymphocytes from Human Peripheral Blood
Authors: NOCERA A. , CADONI A. , ZICCA A. , DI PRIMIO R. , LEPRINI A. , FERRARINI M.
Published: June 2006

Large granular lymphocytes (LGL) are nonadherent cells with cytoplasmic azurophilic granules, avid receptors for the Fc portion of IgG, and a paranuclear localization of alpha‐naphthyl acid esterase or acid phosphatasc. LGL constitute the bulk of TG cells (cells with receptors for sheep erythrocytes and for IgG molecules) and null cells (non‐T, non‐B cells). In the present study we demonstrate that 20–33% of the circulating human LGL express receptors for the third complement component (C3R). When TG cell or null cell fractions from normal individuals or non‐T cells from a patient with infantile agammaglobulinaemia (which contained almost exclusively LGL) were rosetted with erythro cytes coated with antibody and complement, a variable number of C3R‐bearing cells were detected. Such cells were isolated and analysed further; the great majority of them displayed the cytochemical and ultrastructural features of LGL.
Release of Mediators from Purified Rat Mast Cells during Phagocytosis
Authors: CZARNETZKI B. M.
Published: June 2006

Purified mature rat peritoneal mast cells, on exposure to zymosan or latex beads, phagocytize these particles, although less efficiently than macrophages. During phagocytosis, histamine, β‐glucuronidase, and eosinophil chemotactic factor are released from mast cells in a time‐, temperature‐ and dose‐dependent fashion. Complement components, cytochalasin B (5 μg/m1, and indomethacin (10‐6m). enhanced mediator release, whereas compound BW 755C (20 μg/ml), a cyclooxygenasc and lipoxygenase inhibitor of arachidonate metabolism, totally abolished this process. Phagocytosis of mast cells thus activates imracellular mechanisms that closely resemble those observed with other phagocytic cells. These observations add a new perspective to the role of mast cells in inflammatory events.
The Effect of C4 on C1 Binding and Activation
Authors: YMAN I. MALMHEDEN , EIM G. STALENH , SJÖQUIST J.
Published: June 2006

Cell‐bound human C4 enhances the uptake of CT by immunoglobulin‐carrying cells. The effect is more pronounced with small amounts of IgG than with large amounts. Furthermore, the presence of C4 on the cell surface dramatically promotes the enzymatic effect of C1 on C7. The C1 binding is efficiently blocked, and the C1 activation is almost abolished by pretreating IgG‐bearing cells with protein A from Staphylococcus aureus. In contrast, protein A has no effect on cells carrying C4 in addition to IgG. Protein A added in small amounts to IgG‐coated sheep erythrocytes inhibits the haemolysis by subsequently added human scrum, whereas even larger amounts of protein A has no effect when added to cells carrying C4 as well. Cells with and without C4 take up protein A equally well.
In Vitro Production of Monoclonal and Polyclonal Immunoglobulins by Peripheral Blood Mononuclear Cells in Human Plasma Cell Myeloma
Authors: PEEST D. , HOLM G. , MELLSTEDT H. , PETTERSSON D.
Published: June 2006

The in vitro monoclonal and polyclonal immunoglobulin (Ig) production of peripheral blood mononuclcar cells was studied in human multiple myeloma (four IgG myelomas, one IgA myeloma) and in one patient with benign monoclonal gammopathy. Using an enzyme‐linked immunosorbent assay with anti‐class‐specific antisera and antisera against idiotypic structures on the myeloma protein, it was possible to quantilate separately monoclonal and polyctonal Ig of the same class in cell culture supernatants. After stimulation with pokcweed mitogen (PWM) patients' cells produced lower amounts of polyclonal Ig than cells from healthy adults. In contrast, production of monoclonal Ig could not be enhanced by PWM. Moreover, the kinetics of monoclonal Ig production was different from that of polyclonal Ig. Myeloma cells contained large amounts of monoclonal Ig while their content of polyclonal Ig was low. A rapid release of preformed monoclonal Ig during the first day of culture was not inhibited by puromycin. A later phase of release was partly suppressed by puromycin and was probably caused by active protein synthesis.
Fcγ‐like Determinants on Immunoglobulin Variable Regions: Identification by Staphylococcal Protein A
Authors: BIGUZZI S.
Published: June 2006

Protein A from Staphylococcus aureus is widely known as a ligand specific for the Fc portion of the IgG molecule. However, recent reports suggest that a minor proportion of immunoglobulins may also carry protein‐A‐reactive sites in their Fab portion. I Found that about 5% of polyclonal Fabγ and Fabμ fragments strongly bound protein A. Moreover, 2 of 18 human IgG myelomas showed a similar reactivity. This interaction was not due to a conventional anti‐protein‐A antibody activity, since it was possible to obtain F(ab′)2, from human IgG anti‐tetanus toxoid, and Fab. from mouse IgG anti‐sheep erythrocytes, which were able to bind protein A and the immunogen. The finding that a proportion of Fv fragments (VH+VL domain), obtained from normal human IgM, also interacts with protein A suggests that the reactive site is located within the framework of the variable portion of a subset of immunoglobulins. The ability of these F(ab′)2 fragments to inhibit efficiently the binding of [1251]protein A to immobilized human Feγ fragment, suggests that these immunoglobulins express, within their variable regions, determinants that possess Fcγ‐like properties.
Characterization of an Lyt‐l+ Cytolytic T‐Cell Clone Specific for a Polymorphic Domain of the I‐Ak Molecule
Authors: PIERRES A. , SCHMITT‐VERHULST A.‐M. , BUFERNE M. , GOLSTEIN P. , PIERRES M.
Published: June 2006

We have characterized a cytolytic T‐cell clone, isolated from an A.TH anti‐A.TL mixed lymphocyte culture, which recognized a private determinant of the I‐Ak molecule. This specificity has been confirmed by inhibition of effector‐target cell interaction by anti‐I‐Ak monoclonal antibody (mAb). Comparison of the inhibitory capacity of various mAb and the spatial arrangement of their epitopes (defined in previous studies by antibody‐binding competition) indicated that the antigenic site recognized by this cytolytic T‐cell clone was topologically related to one of the major polymorphic domains of the Ak molecule. This clone expressed the Thy‐1.2+, Lyt‐1.2+, Lyt‐2+low and I‐AS‐ cell surface phenotype. Testing of several rat mAb, screened for their ability to inhibit H‐2K/D‐specific cytolysis at the level of the effector cells, revealed that two anti‐p94, 180 mAb but not various anti‐Lyt‐2 mAb inhibited the lytic function of this anti‐I‐Ak T‐cell clone.
Role of the Ek Molecule in the Generation of Suppressor T Cells in Response to LDHB
Authors: BAXEVANIS C. N. , ISHII N. , NAGY Z. A. , KLEIN J.
Published: June 2006

The role of the Ek (EαEβk) molecule in the generation of suppressor T (Ts) cells specific for lactate dehydrogenase B (LDHB) was studied using different approaches. First, lymph node cells from LDHB‐primed B1O.A(2R) (AkEk) nonresponder mice were shown to suppress the LDHB‐specific and Ak‐restricted proliferative response of T cells from the congenic responder strain B10.A(4R), which does not express E molecules (AkEo). Similarly, lymph node cells from primed CBA (AkEk) mice suppressed the anti‐LDHB response of Lyt‐1+Lyt‐2‐ T celfs (depleted of Lyt‐2‐bearing TS cells) from the same mice. Second, in vitro priming of 2R (AkEk) T cells with LDHB‐pulsed 4R (AkEo) antigen‐presenting cells (APC) generated T‐cell proliferation but not suppression. Third, nonresponder 2R mice were turned into responders by injecting them with LDHB‐pulsed 4R APC or monoclonal Ia.m7 antibody that blocks the Ek molecule. The data demonstrate that expression of Ek molecules by the APC is necessary to generate LDHB‐specific Ts cells, which in turn prevent the proliferation of Lyt‐l+Lyt‐2‐ (probably helper) cells recognizing the same antigen in the context of the Ak molecule.
Cyclosporin A Inhibits Thymus‐dependent but not Thymus‐independent Immune Responses Induced by Dextran B512
Authors: FERNANDEZ C. , PALACIOS R. , MÖLLER G.
Published: June 2006

We have compared previous studies of the immune response to dexlran (Dx) B512 in thymus‐deficient nu/nu and thymectomized, lethally irradiated, and bone marrow‐reconstituted mice with those obtained with cyclosporin A (CyA) as a T‐cell‐inhibiting drug. Our data show that only immune responses to TD forms of Dx B512 are susceptible to suppression by CyA, whereas anti‐alpha 1–6 and anti‐DNP antibodies induced by high molecular weight dextran and DNP‐native Dx, respectively, were not inhibited. Similar results were obtained when polyclonal responses were studied. The polyclonal response induced by another dcxtran preparation, the polyanion dextran sulphate (DxS), was inhibited by CyA to the same extent as polyclonal T‐cell activation by T‐cell mitogens.
Glucocorticosteroids in Renal Transplantation
Authors: HÄYRY P. , VON WILLEBRAND E. , AHONEN J. , EKLUND B.
Published: June 2006

We have made an attempt to define a more ‘economical’ method for glucocorticosteroid (GS) administration in renal transplantation in man by shifting most of the GS to the immediate post‐operative period. Thirty cadaver kidney recipients were prerandomized in 2 groups of 15 patients and monitored by transplant aspiration cytology (TAC). After transplantation the high‐dose experimental group received 3.5 mg/kg/day of methyl‐predinisolone (MP), tapered to 0.4 mg/kg/day in 14 days, and the low‐dose control group 1.0 mg/kg/day, tapered to 0.4 mg/kg/day in 5 days. Upon rejection the patients in the high‐dose experimental group were treated with at most 7 mg/kg/day of oral MP divided into four doses daily, whereas the low‐dose control group received 1–3 intravenous boluses of 14 mg/kg (approximately 1000 mg/70 kg). Within 30 days after the transplantation, 7 clinically manifest rejection episodes and 2 episodes of in situ inflammation (without signs of rejection) were recorded in the experimental group, compared with 18 clinically manifest rejection episodes and 6 episodes of inflammation in the control group. The onset of the first episode of inflammation was delayed from 5.0±1.5 days in the control group to 6.6 ± 1.0 days (P=0.00) in the experimental group, and the onset of the first clinical rejection from 5.4±2.1 days to 8.5±1.0 days (P=0.00), respectively. TAC analysis documented that the first episode of inflammation was significantly smaller in the experimental group than in the control group (P=0.001), although the frequency of T lymphoblasts and monocytes was similar to that in the control group (P= 1.00 and 0.20), the frequency of in situ B lymphoblasts and lymphocytes was moderately depressed (P=0.04 and 0.04), and the accumulation of macrophages was both depressed (P=0.07) and delayed. After high initial MP administration the rejections were shorter and easier to overcome: the duration of the first inflammation episode was reduced from 10.5±4.4 days in the control group to 5.9±4.0 days (P=0.02) in the experimental group and the duration of clinical rejection from 8.4±4.1 days to 3.9±3.4 days, respectively (P=0.01). After 18‐month follow‐up study 12 patients in the experimental group had a life‐supporting graft, compared with 7 patients in the control group. There was no difference in the frequency or type of complications between the two groups.
Autologous Mixed Lymphocyte Reaction in Man
Authors: DAMLE N. K. , GUPTA S.
Published: June 2006

Human peripheral T cells and T‐cell Subpopulations defined with monoclonal antibodies of the OKT series were studied for the proliferative response on stimulation with autologous non‐T cells or mitogen‐activated T cells as stimulators in autologous mixed lymphocyte reaction (AMLR). T cells exhibited a vigorous proliferative response when stimulated with autologous non‐T cells or activated T cells but not with unactivated T cells. The stimulatory capacity of non‐T cells or activated T cells in AMLR was inhibited by prior treatment of stimulator cells with 7, 2 anti‐human Ia framework‐specific monoclonal antibody in the absence of complement. A BUdR and light technique was used to ablate proliferating T cells in response to either non‐T cells or activated T cells in AMLR. The removal of T cells responding to autologous non‐T cells left the responsiveness to autologous activated T cells relatively intact. Conversely, the removal of T cells responding to autologous activated T cells left the responsiveness 10 autologous non‐T cells relatively intact. Thus T cells responding to autologous non‐T cells appear to be distinct from those responding to autologous activated T cells in AMLR. Further analysis with OKT4 and OKT8 monoclonal antibodies showed that the major population responding to autologous non‐T cells was contained in OKT4+ T cells and that responding to autologous activated‐T cells was contained in OKT8+ T‐cell subset. However, both OKT4+ and OKT8+ T‐cell subsets responded by proliferation to non‐T cells and activated T cells in allogeneic MLR. T cells selected after AMLR between T and non‐T cells or T and activated T cells were treated with mitomycin and examined for their regulatory influence on the proliferative response of fresh autologous responder T cells. T cells selected from T and non‐T AMLR augmented and those obtained from T and activated T AMLR suppressed the proliferative responses of fresh autologous T cells to phytohaemagglutinin and to autologous or allogeneic non‐T cells in AMLR or allogeneic MLR. These findings indicate that in AMLR between T and non‐T cells or T and activated T cells phenotypically distinct Subpopulations of T lymphocytes respond by proliferation and express distinct immunoregulatory functions.
High Incidence of Spontaneous Ig‐Producing Lymphocytes in Peripheral Blood and Synovial Fluid of Patients with Active Seropositive Rheumatoid Arthritis
Authors: AL‐BALAGHI S. , STRÖM H. , MÖLLER E.
Published: June 2006

Numbers of in vitro spontaneous IgG, IgM and IgA plaque‐forming cells (PFC) as assessed by a modification of the protein A haemolytic plaque assay were determined in the blood and synovial fluid of patients with seropositive rheumatoid arthritis (RA) and compared with those of control groups. The total numbers of PFC were significantly higher in the peripheral blood of patients with active seropositive RA than in that of normal controls. In addition, most B lymphocytes in the synovial fluid of patients with active seropositive RA were active immunoglobulin (Ig) producers, whereas synovial fluid lymphocytes from patients with inactive seropositive RA and seronegative arthritis were not. In general, IgA PFC were relatively high in blood, whereas IgG PFC dominated in the synovial fluid. IgM PFC appear to be relatively low in blood and synovial fluid. However, a relative increase of IgG PFC was noted in the peripheral blood of patients with active RA. To test for polyclonality of the increased Ig synthesis, we tested the sera of patients and controls for the presence of polyclonal antibodies against sheep erythrocytes (SRBC) and SRBC modified by fluorescein isothiocyanate (FITC) and trinitrophenyl (TNP). No differences were observed with SRBC and TNP‐SRBC agglutinin titres between patients and controls, but patients with RA had higher titres of FITC‐SRBC agglutinins than normal sera. This finding supports the concept of a polyclonal nature of antibody production in RA patients.
The Immunological Reactivity of the Three Homologous Repetitive Tetrapeptides in the Region 41–64 of Allergen M from Cod
Authors: ELSAYED S. , SORNES S. , APOLD J. , VIK H. , FLORVAAG E.
Published: June 2006

The CD‐calcium binding loop of Allergen M encompasses the two homologous tetrapep‐tides Asp‐Glu‐Asp‐Lys and Asp‐Glu‐Leu‐Lys. These tetrapeptides were recently suggested to be mutually critical for the immunological specificity of this region. To obtain evidence of the validity of this hypothesis, four peptide analogues encompassing these tetrapeptides at the termini were produced by manual solid‐phase peptide synthesis. The structures of these peptides were ascertained by amino acid analyses and end‐group sequence determination. The crude preparations were prepared by high‐performance liquid chromatography and high‐voltage electrophoresis. The immunological reactivity was investigated by using the preliminary purified peptides in the IgE and IgG test systems used. The results of radioallergosorbent test inhibition showed that all the synthetic peptides encompassing a minimum of two of the tetrapeptides have the capacity to bind specific IgE of the serum pool. The composition and sequence of the six spacer amino acids between these tetrapeptides were of no importance for the acitivity. The highest inhibition percentage was obtained by the peptides derived from the primary structure of Allergen M. No activity was found for the control peptides synthesized under identical conditions. Similar results were shown by PIC tests, using two cod‐allergic sera and two recipients. In the IgG test system, all the four peptides containing the two terminal tetrapeptides showed an antigenic reactivity. They deflected allergen M line in rocket line immunoelectrophoresis. No deflection was given by the control peptides or proteins used. We conclude that the immunological reactivity of region 41–64 of allergen M is determined by 3 homologous tetrapeptides, repeated in three sites interspaced by six amino acids in a segment of 24 residues.
Localization of T‐Lymphocyte Areas in Human Lymphoid Tissues by Fluorochrome‐Labelled Helix pomatia A Haemagglutinin
Authors: ZEROMSKI J. , HAMMARSTRÖM S. , HELLSTRÖM U. , BIBERFELD P. , PERLMANN P.
Published: June 2006

Fluorochrome‐labelled Helix pomatia A haemagglutinin (HP) stained T‐dependent areas in cryostat tissue sections of human lymph node, tonsil, and spleen and thymocytes within thymic medulla but not within thymic cortex. Neuraminidase treatment of the tissue section was a prerequisite for the positive cell staining. The staining was cell‐membrane‐associated, and the histological pattern was essentially the same as that observed in parallel sections treated with monoclonal antibodies to T cells (OKT3, Leu‐I). In addition, blood vessel walls and connective tissue were also stained by HP conjugates. The latter staining was independent of neuraminidase treatment, resistant to α‐galactosidase treatment and was not blocked by pretreatment with anti‐fibronectin antibody. Both types of staining reaction were specific inasmuch as unlabelled HP and the competitive sugar hapten N‐acetyl‐D‐galactosamine blocked the reaction. Frozen, formalin‐fixed tissue sections showed the same stamability and reaction pattern as unfixed sections.
Effect of Cyclosporin A on the in Situ Inflammatory Response of Rat Renal Allograft Rejection
Authors: NEMLANDER A. , SOOTS A. , VON WILLEBRAND E. , TALLQVIST G. , HÄYRY P.
Published: June 2006

The impact of cyclosporin A (CyA) on a normal kidney parenchyma and on the in situ inflammatory response of rejection was investigated in normal DA rats and after transplantation of DA renal allografts to Lewis recipients. In a normal, non‐transplanted DA kidney more than 80 mg/kg/day of CyA induced light‐microscopic changes in the distal tubular cells of the renal cortex and outer medulla. These changes were not accompanied by any visible inflammation and were directly proportional to the dose of the drug and to the duration of drug administration. Treatment of a transplant recipient with 40 mg/kg/day of CyA abolished or at least efficiently reduced the in situ inflammatory response of rejection both as analysed from tissue sections and as quantified from the recovery of inflammatory cells after enzymatic digestion. It also reduced efficiently not only the number of T and B blast cells of the inflammatory infiltrate but also the number of other inflammatory cells, such as in situ lymphocytes, monocytes, and macrophages, and abolished or at least reduced the generation of (T) killer cells in situ and in the recipient spleen. These effects were inversely proportional to the time elapsed between grafting and initiation of treatment: although a complete suppression of all three features was obtained if the drug treatment was initiated already on the day of transplantation, a significant reduction of these functions was still found if the treatment was initiated later when the blastogenic response was already underway.
In Vitro Stimulation of Lymphocytes in Leprosy Patients, Healthy Contacts of Leprosy Patients, and Subjects Not Exposed to Leprosy
Authors: CLOSS O. , REITAN L. J. , NEGASSl K. , HARBOE M. , BELEHU A.
Published: June 2006

In vitro lymphocyte stimulation was performed on peripheral blood lymphocytes from 48 leprosy patients, 15 healthy contacts of leprosy patients, and 16 normal controls who lived in a leprosy‐free area and who had not been exposed to leprosy. Tuberculin PPD and an antigen fraction, MLW 1, prepared from M. leprae, were used as stimulants. The MLW 1 preparation contained one antibody‐precipitable component when tested in crossed immuno‐electrophoresis against a polyvalent anti‐M. leprae immunoglobulin preparation, namely the ML 7 antigen. MLW 1 induced strong lymphocyte responses in patients with tuberculoid leprosy and healthy contacts of leprosy patients, but only weak or no responses in lepromatous leprosy patients and non‐exposed controls. A marked depression of the response to tuberculin PPD was observed in lepromatous leprosy patients. The specificity of the MLW 1 antigen is discussed, and a new estimator of specific lymphocyte stimulation, the δcpm′, is introduced.
A Comparative Analysis of Cell Surface Markers on Murine NK Cells and CTL Target‐Effector Conjugates
Authors: BEAUMONT T.J. , RODER J. C. , ELLIOTT B. E. , KERBEL R. S. , DENNIS J.W. , KASAI M. , OKUMURA K.
Published: June 2006

The rosetting of sheep erythrocytes (SRBC) coated with non‐haemagglutinating monoclonal antibodies rather than conventional haemagglutinating antisera revealed readily detectable FcR on most splenic natural killer (NK) cells since 76% of splenic lymphocytes forming conjugates with YAC also resetted with SRBC coated with high concentrations of monoclonal anti‐SRBC antibody of the IgG2b subclass and since Ficoll depletion or enrichment of splenic lymphocytes rosetting with IgG2b‐coated SRBC resulted in a corresponding 4‐fold decrease or increase in conjugate‐forming cells and a 10‐fold decrease or increase in NK cytolytic acttvity. NK cells bound much less readily to monoclonal IgG2a and not at all to monoclonal IgGI or IgM, but the degree of binding was directly proportional to the amount of antibody on the erythrocytes and was not isotype‐restricted. In addition, immunofluorescent studies revealed that YAC‐1‐conjugated lymphocytes were Lyt‐1‐, Lyt‐2‐, partially Thy‐1+ (60%), asiato‐GMI + (80%), Qa‐4+ (77%), Qa‐5+ (79%), and Ly‐5+ (94%). In comparison, a proportion (39%) of alloimmune peritoneal exudate cells which conjugated with P815–2 also siained by immunofluorescence with anti‐asialo GM1 antisera. Most (>90%) P815‐ conjugated cells were Thy‐1+, Lyt‐2+. and a subpopulation of Lyt‐l+2+ conjugates was observed (25 %). Qa‐5 and Ly‐5 were also expressed on most (two‐thirds) cytolytic T lymphocytes (CTL) conjugates, whereas Qa‐4 and FcR for IgG2b were not detected. The best phenotypic distinctions between NK cells and CTL were therefore based on the presence or absence of Lyt‐2, Qa‐4, and FcR for IgG2b on most effector cells. Anti‐asialo‐GMl or monoclonal anti‐Qa‐4 and complement treatment greatly diminished both the frequency of NK conjugates and the percentage of conjugates with detectable IgG2b FcR or asialo‐GM1. These results confirm that NK cells co‐express asialo‐GMI and Fc receptors, at the single‐celt level, and provide a simple method for greatly enriching NK populations at least 10‐fold.
Effect of Cyclosporin A on the in Situ Inflammatory Response of Human Renal Allograft Rejection
Authors: HÄYRY P. , AHONEN J. , VON WILLEBRAND E. , EKLUND B. , HÖCKEKSTEDT K. , KAUSTE A. , TASKINEN E. , LAUTENSCHLAGER I. , LALLA M. , SARELIN H.
Published: June 2006

Twenty cadaveric kidney allograft recipients were prerandomized into two groups. Ten patients (control group) were treated postoperatively with azathioprine (AZA) plus methylprednisolone (MP); the other ten received cyclosporin A (CyA) as the only immunosuppressive agent. Both groups received MP during rejection. One patient was excluded from the CyA group because of an early postoperative cardiac infarction and death. All transplants were monitored by alternate‐day fine‐needle aspiration biopsy and transplant aspiration cytology. Some patients treated with CyA had a significant initial decrease in urine output, reaching control values approximately 1 week postoperatively. The mechanism behind this deteriorated renal function is not clear, but it seemed to have been caused by injuries to the kidney tubular component, since a distinct monocytic‐lymphocytic inflammation and severe cytological changes resembling pronounced acute tubular necrosis were observed concomitantly in transplant aspiration cytology. The CyA‐treated patients had normal levels of blood leucocytes, thrombocytes and lymphocytes but displayed a strong early blood eosinophilia that was absent in the control subjects. During the first 30 days after transplantation 15 in situ episodes of inflammation were recorded in the nine transplants treated with CyA, whereas only 6 episodes were found in the 10 transplants receiving AZA + MP (P<0.01). The first inflammatory episode in the CyA‐treated transplants peaked between days 5 and 8 after transplantation and was followed by another distinct inflammatory episode between days 23 and 26. In the AZA‐ plus MP‐treated transplants, only one inflammation episode was observed, with a peak on day 14 postoperatively. The inflammatory cell types most prominently present in the CyA‐treated transplants were lymphocytes, B plasmablasts and monocytes. The early inflammatory episodes in the CyA‐treated transplants may have been related to the fact that during the initial intramuscular administration, therapeutic CyA concentrations in patient serum were not achieved until the fourth postoperative day during peroral administration. The onset of transplant function had no impact on the in situ inflammatory response of rejection in the CyA‐treated transplants or on the concentration of CyA in patient serum. This indicates that CyA may also be used in initially non functioning transplants. Confirming our earlier results, we were unable to demonstrate the major histocompatibility complex (MHC) antigens on the healthy grafts treated with AZA plus MP. However, in healthy allografts treated with CyA, both classes of MHC antigens were nearly invariably demonstrable on the graft endothelial cell surface. Approximately 60% allograft survivals were recorded in both groups at 6 months, and all patients with functioning grafts were able to work.
Induction of Immunological Tolerance to Single‐stranded and Double‐stranded DNA
Authors: SASAKI T. , KADONO T. , ENDO F. , ISHIDA S. , YOSHINAGA K.
Published: June 2006

Single‐stranded (ss) and double‐stranded (ds) DNA were conjugated with the copolymers of d‐glutamic acid and d‐lysin (d‐GL). Administration of ss‐DNA‐d‐GL conjugates to C3H/He and NZB/W F1 mice could render the mice tolerant to both direct and indirect anti‐ss‐DNA antibody‐forming cell responses, irrespective of their immune status. Repeated administration of ss‐DNA‐ or ds‐DNA‐d‐GL conjugates decreased the levels of anti‐ss‐DNA and anti‐ds‐DNA antibody titres and reduced the occurrence of ss‐DNA and ds‐DNA antibody‐forming cells even in old female NZB/W F1 mice that had already developed an autoimmune state with lupus nephritis. The unresponsiveness was DNA‐specific, and the state of tolerance was stable in vitro at the cellular level after the removal of the tolerogen. This tolerance model would be useful in analysing the regulatory mechanisms in anti‐ss‐DNA and anti‐ds‐DNA antibody production, and application of this kind of therapy in the treatment of systemic lupus erythematosus is suggested.
Abnormalities of Immunocompetent Cells in Primary Biliary Cirrhosis
Authors: RAMARLI D. , CANONICA G. W. , MELIOLI G. , BIAGIONI P. , NIERI S. , CASINI A. , SURRENTI C.
Published: June 2006

Primary biliary cirrhosis (PBC) is a chronic cholestatic progressive liver disease frequently associated with various immunological abnormalities. We have studied the influence of normal and PBC adherent cells (AC) on peripheral blood lymphocytes (PBL) colon) growth both in liquid and soft agar culture. Co‐culture experiments with different combinations of AC and PBL of PBC patients and normal donors provided evidence that PBC adherent cells function abnormally. However, this impaired function is not the sole factor involved, since the number of colonies formed by PBC lymphocytes in the presence of normal AC was twice the normal value. The possibility that this increase is correlated with the state of activation of T cells is discussed in the light of the high number of DR‐positive T cells found in the PBC patients studied. Finally, none of these immunological abnormalities was related to the stage of the disease.
Development of a Serum‐Free Defined Culture Medium for Lymphoblast Transformation Tests of Mouse Spleen and Thymus Cells
Authors: KRISTENSEN F. , WALKER C. , WÄLTI M. , DE WECK A. L.
Published: June 2006

By analysis of thymidine uptake during the first 4 h of incubation and by examining the responsiveness of unfractionated mouse spleen cells upon mitogenic or allogeneic stimulation, some serum factors of major importance for in vitro cultivation of lymphocytes have been examined. Albumin and L‐alanine are essential for the maximal preservation of the in vivo‐initiated lymphocyte activity during the first hours of incubation in vitro. Transferrin plays a major role in the lymphocyte proliferation, induced by mitogens in vitro, and, finally, zinc and selenium exert a clear enhancing effect on the response to an allogeneic stimulation. The AATSZ medium (RPMI 1640 enriched with t‐alanine, albumin, transferrin, zinc chloride, and sodium selenite) enables a proliferation of the same magnitude as or higher than PCS medium. The kinetics are the same, and the cell viability is comparable, but standardization is much simpler with AATSZ. This is primarily because FCS binds some mitogens and contains inhibitors. Consequently, the standardization of such a culture system is dependent on variations from serum batch to serum batch. On the other hand, the current composition of the AATSZ medium promotes the sticking capacity of T lymphocytes and does not support growth of all lymphoid cell lines. Consequently, this defined medium, although not yet suitable as optimal medium for all lymphocyte functions, can advantageously be used for short‐term studies of most murine lymphocyte functional and cooperation studies in vitro.
Affinity and Inhibitory Capacity of T‐Cell Proliferation of Monoclonal Anti‐Ia Antibodies
Authors: LEMONNIER F. A. , BIRNBAUM D. Z. , DUBREUIL P. C. , CAILLOL D. H.
Published: June 2006

The rates of dissociation from the surface of la‐positive spleen cells of three different anti‐Ia monoclonal antibodies were evaluated and compared with their inhibitory effects on T‐ lymphocyte proliferate responses involving the same spleen cell population, either as a source of accessory cells (responses to soluble antigens) or as allogeneic stimulating cells. In general, a direct relationship was observed between the affinity with which these monoclonal antibodies interact with Ia‐positive cells and the magnitude of their inhibitory effects, whether soluble antigen‐ or alloantigen‐induced responses were tested. These results indicate that the affinity of monoclonal antibodies is a factor that has to be considered to the same extent as their fine specificity when interpreting the results of inhibitory studies of T‐cell responses by anti‐Ia monoclonal antibodies.
In Vivo Collaboration between Precursor T Cells and Helper T Cells in the Development of Delayed‐Type Hypersensitivity Reaction to Influenza Virus in Mice
Authors: LEUNG K. N. , SCHILTKNECHT E. , ADA G. L.
Published: June 2006

Nude, athymic mice do not mount a delayed‐type hypersensitivity (DTH) response to influenza A virus. A single injection of T helper cells (γ‐irradiated, 2‐day immune spleen cells) or three injections over 3 days of a concanavalin‐A‐activated spleen cell supernatant to virus‐sensitized nude mide resulted in a ‘normal’ DTH response when the mice were challenged with the virus. It was previously shown that the cells responsible for the reaction were T cells and required I‐region compatibility. Injection of T helper cells into normal mice did not affect the level of the subsequent DTH response. However, injection of such cells into mice pretreated with anti‐thymocyte serum (ATS) restored the ability of the mice to mount a DTH response. The results show that (1) nude mice contain precursor T cells for influenza virus antigen; (2) an I region‐restricted response can be generated in the absence of a thymus; and (3) in vivo collaboration between DTH T‐cell precursors and helper T cells can be shown to occur in congenitally nude mice and ATS‐treated mice.