bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection and Quantification of the Terminal C5b‐9 Complex of Human Complement by a Sensitive Enzyme‐Linked Immunosorbent Assay
Authors: MOLLNES T. E. , LEA T. , HARBOE M.
Published: June 2006

An enzyme‐linked immunosorbent assay for detection and quantification of the terminal complexes (SC5b‐9 and membrane attack complex) of human complement is described. We separate the complex from the native complement components, to use antibodies against the native components in a ‘double‐antibody sandwich’ technique. It is thereby possible to detect the terminal complement complex in solution without the requirement of specific antibodies against the neoantigens. The results show that the assay is both sensitive and specific. Evidence is presented that a terminal complement complex occurs in a normal plasma pool. The terminal complement complex may be valuable for evaluating both the physiology and palhophysiology of the complement system in vivo.
A Quantitative Assay Detecting Small Numbers of Effector Helper T Cells, Regardless of Clonal Specificity
Authors: POBOR G. , BANDEIRA A. , PETTERSSON S. , COUTINHO A.
Published: June 2006

We have developed a new assay for quantitative detection of all helper T cells that can induce normal B lymphocytes to proliferation and Ig secretion. To establish the optimal assay conditions, we have used cloned T helper cells of defined specificities that had previously been shown to activate normal B lymphocytes expressing the specific antigen(s) on direct cellular interactions. As shown in this paper, ‘irrelevant’ B lymphocytes—that is, those that do not express either antigen or restriction elements recognized by the effect or helper T cells—can also be induced in the presence of appropriate concentrations of pokeweed mitogen which are not mitogenic for the ‘target’ Blymphocytes ‘Nonspecific’ plaque‐forming cell responses are of the same magnitude as those provided to specifically triggered targets and equal or better than those induced by lipopolysaccharide. The assay is highly sensitive and enables ‘semiquantitative’ detection of less than 20–30 effector T cells per culture. Since effector helper T cells can be detected regardless of the clonal specificity, the assay appears useful for quantitative studies of various populations of T helper cells displaying mixed specificities and for the classification of cells with unknown functions.
Detection of Tumour‐Associated Transplantation Antigens by Syngeneic Cytotoxic T Lymphocytes after Fusion of Tumour Cell Membrane Vesicles to Recipient Cells
Authors: ALTEVOGT P.
Published: June 2006

Sendai virus envelope‐mediated fusion was used to transfer plasma membrane vesicles of ESb lymphoma cells (H‐2d) to syngeneic or allogeneic recipient cells. After fusion, ESb‐specific cell surface determinants were detectable on the recipient cells, as revealed by indirect immunofluorescence and cytofluorographic analysis. Fused cells were also tested as targets in 51Cr‐release assays, using syngeneic DBA/2 anti‐ESb‐ or anti‐H‐2d‐specific T lymphocytes (CTL). Fused recipient cells became susceptible to lysis by both types of CTL, indicating that both tumour‐specific and H‐2d determinants were inserted on fusion. The experimental approach presented could be of help as an assay system for the molecular identification of tumour‐specific transplantation antigens.
Characterization of a Subset of Human Natural Killer Cells That Express OKM1 But Lack HNK‐1 (Leu‐7) Antigens
Authors: MAROLDA R. , TILDEN A. B. , ABO T. , DOUGHERTY P. A. , BALCH C. M.
Published: June 2006

The HNK‐1(Leu‐7) monoclonal antibody selectively identifies a population of human granular lymphocytes with natural killer (NK) cell activity. We previously reported that the HNK‐l+ cell fraction purified from blood mononuclear cells accounted for virtually all NK activity in six individuals. In this study we analysed additional normal individuals and found that in eight out of 14 donors HNK‐1+ cells, purified with a fluorescence‐activated cell sorter (FACS). exhibited greatly enriched NK cell activity, whereas HNK‐1 cells did not have significant activity. In four donors the HNK‐1+ cells were enriched in NK activity compared with HNK‐1‐ cells; however, the HNK‐1‐ cells also had moderate levels of activity. In the two remaining donors. NK activity was not enriched in the HNK‐1+ fraction in comparison with the HNK‐1‐ fraction. To determine the cell type responsible for NK activity in the HNK‐1 subset, these cells were further sorted with the FACS into OKM1+ and OMK1‐ tractions and analysed for morphology and function. HNK‐1‐ OKM1‐ cells were found to he small‐ to medium‐sized lymphocytes devoid of NK activity in all donors tested, whereas most HNK‐1‐ OKM1‐ cells were granular lymphocytes and in some donors demonstrated NK function at a level comparable to HNK‐1+ cells. Thus some individuals have an important subset of granular lymphocytes with NK‐cell activity and the HNK‐1‐ OKMl+ phenotype. It is important to account for these cells in studies involving granular lymphocytes and NK cell function.
Specificity of Plasma Cells in the Rheumatoid Synovium
Authors: YOUINOU P. Y. , MORROW J. W. , LETTIN A. W. F. , LYDYARD P. M. , ROITT I. M.
Published: June 2006

Plasma cells synthesizing rheumatoid factors (RF) were identified by fluorescent staining of sections of synovium and macrophage‐depleted cells from dispersed synovial tissue. The latter avoided problems related to sampling errors in studying tissue sections and in the uncertainty raised by the staining of macrophages with intracellutar complexes. Plasma cells producing IgG predominated, and seropositive patients had a higher proportion of IgM producers than seronegative subjects. None the less, in both groups of patients more than 90% of the IgM plasma cells were synthesizing RF. whereas the corresponding figure for IgG was between 50% and 60% Only around 10% of IgA plasma cells were positive for RF. The high percentage of IgM plasma cells making RF would lend to argue for an IgG‐specific response and against direct polyclonal activation as the stimulus. The percentage of IgG‐producing cells positive for RF is also consistent with a dominant response to IgG. Accepting the difference in the relative proportion of total IgM‐ to IgG‐producing plasma cells in seropositive as against seronegative patients, the close similarity between the two groups in the fraction of cells making RF favours Ihe view that the two groups have a comparable underlying immunopathology dependent on IgG autosensitization. From the technical standpoint, the dispersed cell method gives results in line with those obtained with sections but which are easier to read, whereas the fluorescent techniques described give clear and reproducible results for the detection of RF of different heavy‐chain isotype.
Detection of Lactoferrin and Myeloperoxidase Release from Single Neutrophils by a Protein A Plaque Assay
Authors: RICHTER J. , OLOFSSON T. , BRITTON S. , OLSSON I.
Published: June 2006

A haemolytic plaque assay was adopted to detect release of lactoferrin and mycloperoxidase (MPO) from single neutrophils. Target erythrocytes coated with protein A were bound as a monolayer by poly‐l‐lysine to the surface of plastic dish. Secreted lactoferrin and MPO induced plaque formation dependent on the reaction with complement and specific antiserum producing lysis of the protein‐A‐coated sheep red blood cells Lactoferrin was found to be released spontaneously from a fraction of neutrophils while MPO was released only after phagocytosis, reflecting different mechanisms for degranulation of MPO‐containing azurophil and lactoferrin‐containing specific granules.
Stimulatory Effect of Immobilized Glycans on Macrophages in Vitro
Authors: BØGWALD J. , GOUDA I. , HOFFMAN J. , LARM O. , LARSSON R. , SELJELID R.
Published: June 2006

Mouse macrophages were cultured on chemically modified plastic dishes. On dishes covered with immobilized glycans, the macrophages were stimulated as judged by increased 14C‐glucosamine incorporation, increased cytostatic and cytolytic capacities and by morphology as seen by scanning electron microscopy. The corresponding soluble glycans did not have the capacity to stimulate macrophages as measured by these criteria. Plastic surfaces covered with polyethylenimine showed stimulation of the macrophages with regard to some of the parameters measured. These results may indicate that the stimulation is a multistep process and that, contrary to earlier findings, it is not a prerequisite for stimulation that the glycan be intracellular. The results support the idea that a fixed steric arrangement of glycans is necessary for the stimulation of macrophages in vitro.
Highly Increased Natural Killer Cell Number and Lytic Activity in the Murine Peripheral Blood and Lungs after Interferon Induction in Vivo
Authors: HURME M. , SILVENNOINEN O. , RENKONEN R.
Published: June 2006

We have studied the effect of interferon in vivo (induced by polyinosinic‐polycytidylic acid (pIC)) on the natural Killer (NK) cell lytic activity and on the numbers of large granular lymphocytes (LGL) and target‐binding cells (TBC) in different lymphoid compartments. One day after the pIC induction the spleen contained two‐ to four‐fold increased lytic activity without a significant change in percentages of TBC and LGL. In the peripheral Hood the lytic activity was 12‐ to 40‐fold higher, and a concomitant clear increase in the number of LGL and TBC was seen. In the lungs the increase in lytic activity was 8‐ to 16‐fold and in the number of LGLs ca. 3‐fold. These results demonstrate that not only the lytic activity of the pre‐existing NK cells but also the total amount of NK cells is clearly elevated in the body, but this could be seen more significantly in other highly NK‐active organs than the spleen
Auto‐Delayed‐Type Hypersensitivity Induced in Immunodeficient Mice withModified Self‐Antigens
Authors: TARCIC N. , BALER R. , NAOR D.
Published: June 2006

X‐irradialed (250 rad), cyelophosphamide‐treated or ATx A miceinjected with syngeneic trinitrophenylated spleen cells (TNP‐SC) and footpad challenged with syngencic lymphoblasts generated delayed‐type hypersensitivity (DTH) responses 24, 48 and 72 h after challenge. The syngeneic‐DTH (syn‐DTM) response was mediated by Lyt‐l+ cells and suppressed withLyt‐1+2+3+, I‐Jk+ cells. The suppressorcells were obtained from spleens or thymuses of normal syngeneic mice. Suppressor factor (SF) was extracted or released from Lyt‐l+2+3+, I‐Jk+ cells obtained from normal A mice (but noi fromX‐irradiated A mice). The factor blocked the DTH responses of X‐irradiated mice injected with syngeneic TNP‐SC and challenged with syngeneic lymphoblasts when injected into the mice both at the induction phase and the elicitation phase of the DTH. The factor failed to abrogate allogeneic and xenogeneic DTH. However, allogeneic factor (derived from C57BL/6 mice) abolished the syn‐DTH response of mice injected with syngeneic TNP‐SC and challenged with syngeneic lymphoblasts, The SF was produced by Lyt‐1+2+3+, I‐Jk+ T cells or by thymocytes. The combined extracted product of Lyt‐l+ and Lyt‐2+ cells did not abrogate the syn‐DTH response. Normal spleen cells depleted of phagocytes by a magnetic procedure also produced the SF. These findings indicate, therefore, that suppressive factor (or factors; see Discussion in the accompanying paper, Ref. 17) controls the immunological autoreactivity against syngeneic TNP‐SC.
Effect of Mouse Passage on Fc Receptor Expression by Group A Streptococci
Authors: REFS K. J. , YARNALL M. , AYOUB E. M. , BOYLE M. D. P.
Published: June 2006

The expression and stability of receptors for the Fc region of human IgG on the surface of group A streptococci was studied. Two strains were sequentially passed in mice 22 times. The Fc‐receptor expression on one group A strain, 529, was unaltered while the expression of Fc receptor on a second, 64, was enhanced and approached the level of Fc‐receptor expression of the protein A‐rich Staphylococcos aureus Cowan strain. The level of Fc‐receptor expression on this organism remained stable for over 18 months of laboratory subculture. Mouse passage did not result in the production of a soluble Fc receptor from either of the streptococcal strains. Heat extraction of the Fc‐receptor‐positive group A strain resulted in solubilization of an Fc‐receptor activity which was functionally distinct from either staphylococcal protein A or the Fc receptor isolated from a group C streptococcus.
Production of Antibodies to Cellular Antigens by EBV‐Transformed Lymphocytes from Patients with Urinary Bladder Carcinoma
Authors: PAULIE S. , LUNDBLAD M.‐L. , HANSSON Y. , KOHO H. , BEN‐AISSA H. , PERLMANN P.
Published: June 2006

Lymphocytes from patients with transitional‐cell carcinoma (TCC) of the urinary bladder were transformed by infection with Epstein‐Barr virus. To obtain B cells secreting antibodies reactive with TCC cells, the transformed cells were either adhered to irradiated monolayers of cultured allogeneic TCC cells or subcultured at limiting dilution. Supernatants from these cultures were tested in a modified enzyme‐linked immunosorbent assay against fixed cells, isolated plasma membranes, or lipid antigens or were tested by antibody‐dependent cellular cytotoxicity (ADCC), Reactions with antigens derived from the serum source were excluded by proper controls. By this approach a majority of the patients tested (7/12) gave rise to cultures producing antibodies recognizing various cellular antigens. The antibody‐containing supernatants from these cultures were usually of high titres and the reactive antibodies of IgM isotype. One culture, which had been selected by repeated adherence to TCC cells, produced antibodies reactive with such cells in ADCC. None of the antibodies investigated detected antigens exclusively present on TCC cells.
Demonstration of a Helper Factor(s) with T‐Cell‐Replacing Activity in Synovial Fluid
Authors: AL‐BALAGHI S. , STRÖM H. , MÖLLER E.
Published: June 2006

Cell‐free synovial fluid (SF) obtained from patients with rheumatoid arthritis contains a helper factor(s) capable of augmenting the generation of plaque‐forming cells (PFC) in pokeweed mitogen (PWM)‐stimulated normal peripheral blood mononuclear cells (PBMC). This helper factor behaves like a polyclonal B‐cell activator, in that it triggers the formation of IgM, IgG. and IgA PFC. However, SF has little or no effect on the proliferation of PWM‐activaled PBMC. Furthermore, SF was capable of replacing T cells for PWM‐induced differentiation but not proliferation of enriched human blood B lymphocytes. No helper factor or T‐ccll‐replacing activity was found in SF from patients with traumatic synovilis. Fractionation of SF containing helper activity on staphylococcal protein A column indicated that the activity is induced by biologically active molecules distinct from materials that preferentially hind to protein A such as IgG immune complexes. We conclude that the present activity has striking similarities to the recently described B‐cell differentiation factor that is produced by specifically activated T‐cell lines in vitro.
In Vivo Biological Effect of Allogeneic Cultured Thymic Epithelium on Thymus‐Dependent Immunity in Athymic Nude Rats
Authors: SCHUURMAN H. J. , VOS J. G. , BROEKHUIZEN R. , BRANDT C. J. W. M. , KATER L.
Published: June 2006

We have extended our previous siudy of induction of T‐lymphocyte immunocompetence in athymic nude rats by cultured epithelium (CTE) of syngencic urigin to that by CTE of allogeneic origin. Immune responsiveness (IgG‐class antibody und delayed‐type hypersensi‐liviiy) after ovalbumin immunization is detectable by ‐1–6 weeks after transplantation. However, the antibody appears at a slower rate when compared with heterozygous immunocompeient littermates. Seven weeks after transplantation phytohaemagglutinm responsiveness of spleen cells is detectable, and in‘Independent areas of lymphoid organs lymphocytes with helper and non‐helper T‐cell phcnotype are presenl, bul at lower levels than those in heterozygous immunocompetenl littermates. Levels comparable to that of immunocompeient rals are reached about 20 weeks after transplantation. Since CTE contains thymocytes. control experiments consisted of transplantation with high numbers of allogeneic freshly isolated thymocytes in alhymic nude ratv These animals showed IgG‐class antibody formation after ovalbumin immunization, but at lower levels than CTE‐treated rats, and were almost negative in T‐cell immunocompetence assessed in the other assays. We conclude that CTE of allogeneic origin induces T‐eell immunocompetence in athymic nude rats to the level of heterozygous immunocompeteni littermates. This study adds to the rationale of CTE transplantation applied in treatment of thymic dysfunction.
2‐Mercaptoethanol‐Dependent Lipopolysaccharide‐Responsive B Cells in Nylon‐Wool‐Fractionated Spleen Cell Preparations
Authors: EASTMAN A. Y. , LAWRENCE D. A.
Published: June 2006

The ability of lipopolysaccharide (LPS) to influence lymphocyte mitogenresponsiveness to 2‐mercaptoethanol (2‐ME) was investigated. 2‐ME was mitogenic for splenocytes only when fetal bovine serum lots containing elevated endotoxin levels were used to supplement (5%) the culture medium. 2‐ME did not stimulate mitosis of nylon‐wool‐fractionated splenocytes (NWSC): however, 2‐ME induced significan proliferation in the NWSC cell preparations whenendotoxin‐deficient culture medium was supplemented with doses of LPS whichalone did not stimulate significant proliferation. The sIg* and Thy‐1‐/Lyt‐l‐ fractions of NWSC, which represented less than 5% of the cells in the‘T‐cell preparations', were almost completely dependent on 2‐ME for proliferation in response to LPS stimulation. These data indicate that nylon‐wool‐fractionated preparations cuntain splenic B cells that incorporate significant amounts of [3H]thymidine when stimulated with LPS and 2‐ME.
High‐Dose Survival in the Lymphocytic Choriomeningitis Virus Infection Is Accompanied by Suppressed DTH but Unaffected T‐Cell Cytotoxicity
Authors: MARKER O. , THOMSEN A. R. , VOLKERT M. , HANSEN B. L. , CLEMMENSEN I. H.
Published: June 2006

Provided that intracerebral inoculation is applied, an increase in the virus dose from 102to 104 LD50 of lymphocytic choriomeningitis virus (LCMV) leads to strikingly reduced mortality. To analyse the background for this autointerferencc, we measured several virologic and immunologic variables in mice infected with these doses of virus. In the high‐dose mice we found generally higher organ virus titres and serum interferon titres than in the low‐dose mice. Since we could demonstrate that virus‐specific T‐cell cytotoxicity in spleen, peripheral blood, and meningeal exudate was similar after intracerebral infection with large and small virus doses, and since the LCMV infection in the brain qualitatively and quantitatively was independent of the size of virus inoculum, the explanation for the survival of the high‐dose animals is obviously not lack of possibilities for interaction between cytotoxic T cells and infected sensitive targets in the central nervous system. On the other hand, high doses of virus caused a clear suppression of the LCMV‐specific delayed‐type hypersensitivity(DTH). In addition, when splenocytes from high‐dose animals were transferred either intravenously or locally into the footpad of newly virus‐challenged mice. DTH was markedly suppressed as compared with the response after transfer of spleen cells from low‐dose mice. We therefore conclude that autointerferencc in the LCMV infection is due to a selective suppression of Td function. Large amounts of persistent virus late after infection with high doses of virus suggest a central role for Td function also in virus clearance. Finally, our results indicate the existence of two subsets of K, D region‐restricted T cells, one mediating cytotoxicity and the other mediating DTH. This possibility is discussed.
Participation of Cyclophosphamide‐Resistant T Cells in Murine Lymphocytic Choriomeningitis
Authors: DOHERTY P. C. , ALLAN J. E.
Published: June 2006

A severe inflammatory process can be induced by transferring lymphocytic choriomeningitis virus (LCMV)‐immune spleen cell populations into cyclophosphamide (Cy)‐suppressed, LCMV‐infected recipients. The extent of cellular invasion can be quantified accurately by counting cells obtained from the cisterna magna. The underlying lymphocyte‐target cell interactions are apparently very specific, with massive cellular invasion being dependent on the donor and recipient sharing class I major histocompatibility complex glycoproteins. Unlike the situation found by others for delayed‐type hypersensitivity in the mouse footpad, the inflammatory process in the cisterna magna was induced to a similar extent by unmanipulated LCMV‐immune spleen cells and by lymphocyte populations from mice that were pretreated with 150–200 mg/kg of Cy. This dose of Cy did not eliminate cytotoxic T lymphocytes (CTL) in the mouse strains used. However, immune spleen cells from Cy‐pretreated mice showed a reduced capacity to clear virus from the brain. Virus persisted even though the inflammatory process resolved. These results support earlier conclusions that immune CTL are directly responsible for the induction of severe meningitis in LCM, although CTL obviously do not constitute the sole effectors involved in eliminating virus from the central nervous system.
Prealbumin in Swedish Patients with Senile Systemic Amyloidosis and Familial Amyloidotic Polyneuropathy
Authors: FELDING P. , FEX G. , WESTERMARK P. , OLOFSSON B. O. , PITKÄNEN P. , BENSON L.
Published: June 2006

A prealbumin (PA)‐like protein was demonstrated in amyloid fibrils both from a patient with senile systemic amyloidosis (SSA) and from a patient in northern Sweden with familial amyloidotic polyneuropathy (FAP). The investigated properties of this protein were similar in the two types of fibrils. The protein had molecular weight, antigenic determinants, and at least one eysteinyl residue in common with the subunit of normal PA. In contrast to normal PA. it contained disulphide‐linked subunits and was to some extent bound to the fibril via the eysteinyl residues. The noncovalent forces between its subunits were weaker than in normal PA. It constituted part of the previously described AScl protein of the SSA fibrils. Proteins with lower molecular weight than the PA monomer were major proteins in both SSA and FAP fibrils. These proteins had similar properties in the two kinds of fibril and may be derived from PA. PA in serum from Swedish patients with FAP and SSA was normal with regard to the isoclectric pH of the monomers and tetramers.
Abnormal Production of and Response to B‐Cell Growth Factor and B‐Cell Differentiation Factor in Patients with Systemic Lupus Erythematosus
Authors: HIROSE T. , HARA M. , KITANI A. , HIROSE W. , NORIOKA K. , KAWAGOE M. , NAKAMURA H.
Published: June 2006

We examined the production of and the response to B‐cell growth factor (BCGF) and B‐cell differentiation factor (BCDF) in 21 patients with systemic lupus erythematosus (SLE) and 23 normal subjects. T cells, 2.5 × 106/ml, were cultured for 24 or 72 h with 1% phytohaemagglutinin (PHA). After absorption of PHA by chicken erythrocytes (CRBC), they were used for BCGF and BCDF. In inactive SLE, BCGF activity was significantly lower than that in normal subjects. Active SLE contained two separate groups, one showing normal BCGF activity and the other showing lower activity than normal. In contrast. BCDF activity from initial culture in active SLE was elevated. The B‐cell response both to BCGF and BCDF was elevated in active SLE without Staphylococcus aureus Cowan I antigen (SAC) preactivation. However, the B‐cell response to SAC was markedly disturbed. Thus SLE B cells were shifted to the mature state in vivo. We also demonstrated pivotal abnormalities of monocytes in SLE B‐cell growth and differentiation. These results may contribute to the understanding of the abormalities of T‐B interactions and the overproduction of antibody in SLE.
Development of Natural Killer Cells: Comparison of the Maturation Rates in Different Lymphoid Compartments
Authors: HURME M. , SIHVOLA M.
Published: June 2006

By means of semisyngeneic bone marrow transplantation, the appearance of donor‐type natural killer (NK) cells in different organs (spleen, blood, lungs) of the recipients was studied. Seven days after the transplantation the first donor‐type NK cells appeared in all these organs, and adult NK levels were reached simultaneously within a couple of days. The first NK cells to appear in every organ divided rapidly (sensitive to hydroxyurea) and contained a high proportion of Thy‐1+ cells. These data suggest that NK cell precursors mature locally and simultaneously in different organs.
Prostaglandin E2 Receptors on Human Peripheral Blood Monocytes
Authors: ERIKSEN E. F. , RICHELSEN B. , BECK‐NIELSEN H. , MELSEN F. , NIELSEN H. KRÆMMER , MOSEKILDE L.
Published: June 2006

The prostaglandin E2 (PGE2) receptor on human peripheral blood monocytes is characterized. The receptor binding at physiological temperature and pH was saturable, specific, and reversible. Scatchard analysis of binding data revealed a linear plot giving a Kd= 1.1 ± 10‐9mol/l and Bmax=4.1 fmol/107 cells, equal to 240 binding sites per cell. PGE2 increased intracellular cyclic adenosine 5′‐monophosphate by a maximal factor of 3. PGF2α and archidonic acid had no stimulatory effects on adenyl cyclase, in accordance with their low binding to the cells. The characterization of the PGE2 receptor on human monocytes creates a basis for the study of the clinical significance of changes in PGE2‐receptor binding in disease states involving PGE2‐monocyte interactions such as various immunological disorders and bone resorption.
Immunological Studies in Acquired Immunodeficiency Syndrome
Authors: HOFMANN B. , DUM N. , PLATZ P. , RYDER L. P. , SVEJGAARD A. , NIELSEN J. O.
Published: June 2006

The lymphocyte transformation response in vitro to mitogens (phytohaemagglutinin, concanavalin A, and pokeweed mitogon) and antigens (purified protein derivative and tetanus) was studied in three patients with acquired immunodeficiency syndrome (AIDS), three patients with pre‐AIDS. and six healthy controls before and after depletion of T4‐ or T8‐positive cells. In controls, TS‐depleted lymphocytes responded as well as peripheral blood mononuclear cells (PBMC) when monocytes were added, whereas T4‐depleted cells gave about 50% of this response to mitogens and no response at all to antigens. No evidence of suppression was seen when various mixtures of T4‐ and T8‐depleted cells were made. In particular, there was a virlually linear relationship between the percentage of T8‐depleted cells and the response to antigens. The PBMC of all AIDS and pre‐AIDS patients had very low or absent responses to mitogens and antigens, and except in one case, this response did not increase after depletion of T8‐positive cells (and addition of monocytes), indicating that these patient cells also lack suppressor activity in this assay. However, a significantly increased response to mitogens was seen when the TS‐depleteu suspensions were adjusted to contain 20,(HK) T4‐positive cells per well, but the response was still significantly lower than that of similar suspensions from controls. Thus, not only are the poor responses of PBMC from AIDS and pre‐AIDS patients due to a low concentration of T4‐positivc cells, but the responsiveness of these cells also seems deficient. Furthermore. TS‐positive patient cells also have an impaired responsiveness. Our experiments do not exclude the possibility that the low response is due to a T8‐negative suppressor cell, but it seems more likely that both the T4‐ and the T8‐positive cells are deficient and/or that there is a deficiency in accessory cells. These possibilities are currently under study.
Characterization of Activities Inducing Polyclonal Maturation of Activated B Lymphocytes to Immunoglobulin Secretion
Authors: LEANDERSON T. , PETTERSSON S.
Published: June 2006

Supernatants from long‐term in vitro‐propagated helper T‐cell clones were analysed for their ability to induce polyclonal maturation of B lymphocytes under conditions that ensured activation and proliferation of the target cells. In gel filtration analysis, three peaks of activity appeared corresponding to molecular weights of about 100 kD, 60 kD, and 30 kD. When the same supernatants were run on sodium dodecyl sulphate gels under reducing conditions, only the last two peaks of activity could be detected. Flat‐bed isoelectric focusing gave two peaks of activity corresponding to isoelectric points of pH 5–6 and pH 8–9. These peaks of activity were devoid of any activity supporting proliferation of either T or B cells, and they induced the expression of several heavy‐chain isotypes. Furthermore, when the same analysis was performed on the B‐cell line WEHI 279.1, only one peak was obtained (molecular weight of about 60 kD; isoelectric point, approximately pH5–6), a result suggesting alternate pathways of induction of B cells to high‐rate secretion cells.
Hapten‐Specific Unresponsiveness in Mice
Authors: HUCHET R. , GRANDJON D.
Published: June 2006

The features of B‐cell tolerance induced in mice with the chemically reactive hapten 2,4, 6‐trinitrobenzenesulphonic acid were investigated after various antigenic challenges. A complete abolition of the IgG response was observed after challenge with trinitrophenol (TNP)‐coupled haemocyanin (TNP‐KLH) associated or not with Escherichia coli lipopolysaccharide. Carrier immunization with KLH (3μg) followed by challenge with TNP–KLH led to a normal IgG anti‐TNP response. However, tolerant mice given 3μg TNP–KLH (instead of KLH) and a repeat injection of TNP‐KLH exhibited a depressed IgG anti‐TNP response. These results are discussed within the framework of tolerance induction in adult mature B cells.
Enriched Langerhans Cells Express More HLA‐DR Determinants Than Blood‐Derived Adherent Cells (Monocytes and Dendritic Cells)
Authors: BJERCKE S. , GAUDERNACK G. , BRAATHEN L. R.
Published: June 2006

When used as antigen‐presenting cells, enriched Langerhans cells (LC) are known to induce a stronger proliferative T‐cell response towards antigens than blood‐derived adherent cells (a mixture of monocytes and dendritic cells). To study the mechanism behind this difference in accessory cell function, we have compared the quantitative expression of HLA‐DR molecules on LC and adherent cells (AdC), using a radioimmunoassay system. The amount of HLA‐DR determinants was calculated to be 50–100 times higher on LC than on AdC.
Cathepsin B Activity in Human Blood Monocytes during Differentiation in Vitro
Authors: MØORLAND B.
Published: June 2006

The activity of cathepsin B was assayed in human blood monocytes during differentiation into macrophages in vitro. Freshly isolated monocytes showed negligible cathepsin B activity. On day 3 in culture the enzyme activity was still very low, but it was markedly increased on day 7, concomitant with the monocytes' morphological differentiation into tissue macrophage‐like cells. A further rise in enzyme activity was seen on day 10 in culture. Acid phosphatase activity showed similar, but less marked, increases in human monocytes during 10 days' culture. Generally, higher enzyme levels were seen in monocytes isolated from buffy coal preparations than from whole blood. Both the level and the rate of appearance of cathepsin B activity were further enhanced by endocytosis of carrageenan in the monocytes. Stimulation with endotoxin from Escherichia coli caused variable enzyme responses, as certain pools of human cells expressed cathepsin B activity compared with controls, whereas others showed no change in activity. Endocytosis of carrageenan had no effect on acid phosphatase activity, whereas stimulation with endotoxin led to increased levels of this enzyme activity in all cultures. The present data suggest that a rise in cathepsin B activity may be a component in the differentiation of human monocytes into macrophages. They further indicate separate regulation of lysosomal enzyme activity in human monocytes after some types of stimulation, as previously shown for mouse macrophages.
Quantification of Soluble E‐Receptor in the Serum of Patients with Various Diseases and Its Accompanying in Vitro Immunosuppression in Neoplasia
Authors: OH SE‐KYUNG , LAPENSON D. , MORGAN A. C.
Published: June 2006

Evidence in the literature indicates that soluble E‐receptor in the serum may modulate T‐dependent immune response. We have developed a solid‐phase radioimmunoassay to measure the soluble form of E‐receptor from various sources. The assay detects solubilized antigens derived from E‐receptor‐bearing T lymphocytes and not with non‐E‐receptor‐bearing B lymphocytes. The sensitivity limit of the assay is 0.1 ng/ml of purified E‐receptor antigen. Using this assay, one can show that the T‐cell mitogen phytohaemagglutinin stimulated both lymphocytes and cells of the resting human T‐cell lymphoma cell line Jurkat to shed or secrete E‐receptor into the culture medium. Results of the radioimmunoassay performed on human sera indicated that some patients with Hodgkin's disease, melanoma, sarcoma, or acute or chronic lymphocytic leukaemia had elevated levels of this antigen in their serum, whereas normal human sera registered lower levels, Elevated levels of the soluble form of E‐receptor in the serum were suggestive of an in vitro assessment of their immunosuppressive activity. These results indicate that activation of T‐cell immunity in vivo may result in humoral immunosuppression.
Naturally Cytotoxic Tonsillar Leukocytes: Phenotypic Characterization of the Effector Population
Authors: CHRISTMAS S. E. , ALLAN G. , MOORE M.
Published: June 2006

Human palatine tonsil sections were examined to investigate the distribution of cells hearing the cell surface markers of peripheral blood natural killer (PB‐NK) cells. Leu‐7+ (HNK‐I+) cells were localized predominantly in lymphoid follicles, whereas OKMI‐, Mac‐1‐, and Mo2‐labelled cells were found in the epithelial and subepithelial regions and epithelial crypts. OKTH10+ cells showed a variable distribution, being found in follicles and interfollicular or subepithelial regions. No. B73,I+ cells could be identified in tonsil section. Leu‐7+ cells appeared not to be responsible for tonsillar natural cytotoxicity, since Leu‐7 (HNK‐1) antibody‐ and complement‐mediated lysis under conditions that markedly reduced PB‐NK activity failed to abolish cytotoxicity, and positive selection by means of the FACS IV gave no enrichment of activity. Similarly, cells labelled with the antibodies B73.1, Leu‐l1b, OKT8, OKT10, and TDK 31.1 (anti‐major histocompatibility complex class II framework determinant) were not enriched with regard to NK activity either. However, positive selection with OKM1, Mac‐1, or Mo2 showed that cells bearing these markers were responsible for essentially all tonsillar NK activity. No large granular lymphocytes were identified in such populations enriched for NK activity. The observation that PB‐NK cells labelled faintly with Mo2 weakens the argument that a non‐adherent mononuclear phagocyte population was responsible for the activity. These data therefore support the existence of heterogeneity within naturally cytotoxic cell populations.
Characterization of the Specificity of Monoclonal Antibodies against Mycobacterium tuberculosis by Crossed Immunoelectrophoresis
Authors: HARBOE M. , COATES A. R. M. , HEWITT J.
Published: June 2006

The specificity of monoclonal antibodies against Mycobacterium tuberculosis was determined by crossed immunoelectrophoresis, Radiolabelled, non‐precipitating monoclonal antibody was incorporated in the top gel together with unlabelled polyvalent, precipitating anti‐bacillus Calmette‐Guérin (BCG) antibody. Inclusion of labelled antibody in individual precipitate fines was demonstrated by autoradiography. Each monoclonal antibody was localized in a single precipitate line: antibody TB73 reacted with BCG antigen 56: TB71 and TB72 reacted with BCG antigen 78: and TB78 reacted with BCG antigen 82. The technique permits precise determination of the specificity of monoclonal antibodies at the level of reactivity with native immunogenic components of complex microorganisms without prior antigen purification.
Cyclosporin A Inhibits Induction but Not Production of Gamma Interferon Induced by Epstein–Barr Virus
Authors: ANDERSSON J. P. , ANDERSSON U. G. , BRITTON S. F. , LEY M. DE
Published: June 2006

Cyclosporin A (CsA) interferes with various T‐cell functions in vitro and is a potent inhibitor of T‐cell‐dependent reactions in vivo, such as graft rejection and control of virus infections. Since human gamma interferon (Hu IFN‐γ) is synthesized by T cells and has a controlling role in regulation of Epstein–Barr virus (EBV) infection, we have studied the effects of CsA on EBV‐induced cellular Hu IFN‐γ release. CsA inhibited dose‐dependently the EBV‐induced Hu IFN‐γ response, studied at the cellular level in human blood lymphocytes. These effects were not due to toxicity of CsA, since at inhibitory levels cellular EBV infection measured as polyclonal IgM production proceeded unaffected. CsA did not affect the number of spontaneous Hu IFN‐γ‐secreting cells, nor did it have any inhibitory effect if added after virus exposure. It is concluded that CsA inhibits induction but not production of cellular Hu IFN‐γ.
Magnetic Monosized Polymer Particles for Fast and Specific Fractionation of Human Mononuclear Cells
Authors: LEA T. , VARTDAL F. , DAVIES C. , UGELSTAD J.
Published: June 2006

Magnetic monodisperse polymer particles were developed and the necessary conditions established to use them for both quantification and fractionation of human peripheral blood mononuclear cell populations. The particles consist of a styrene divinylbenzene core into which magnetite has been deposited by an in situ oxidation process. Thereafter the core has been coated with a hydrophilic polymer containing epoxy and hydroxyl groups. The particles have strong nonspecific binding capacity for protein and can he coaled with the appropriate antibodies by physical adsorption only. However, the hydroxyl groups on the outer polymer also make covalent coupling possible. After appropriate blocking they can be used in a rosette assay for quantification of mononuclear leukocytes previously sensitized with monoclonal antibodies. Furthermore, a suitable magnet makes it possible to deplete the cell suspension efficiently of the rosette‐forming cells. We have thoroughly investigated the functional properties of human mononuclear cells depleted of T lymphocytes by this technique. Our results show that T cells are virtually completely eliminated, as demonstrated by flow cytometry and various functional assay systems.
Cyclosporin A Inhibits a Discrete Step in T‐Lymphocyte Stimulation
Authors: LANDEGREN U. , WIGZELL H.
Published: June 2006

Cyclosporin A (CyA) interferes with immune responses and prevents growth Factor release by stimulated T cells. However, it is not known whether this is due to an effect on the accessory cells, required for T‐cell responses, whether antigen recognition cannot occur, or whether later steps, leading to lymphokine production, are blocked. For this reason, the effect of CyA on homogeneous populations of T tumour cells was investigated. The immunosuppressive compound efficiently prevented T‐cell growth factor (TCGF) (interleukin 2) release by stimulated tumour cells. Still, the cells retained the surface antigen T3, known to be involved in T‐cell stimulation, after treatment with CyA. Furthermore, CyA failed to affect the inhibition of proliferation, observed in a T‐cell tumour in response to stimulation, indicating that the cells had received the stimulatory impetus. TCGF release, induced by treatment with a phorbol ester, was only partly sensitive to inhibition by CyA, demonstrating that CyA will interfere with discrete aspects of the stimulation of a T‐cell.
Fibronectin Binds to Complement‐Coated Agarose Beads and Increases Their Association to Mouse Macrophages
Authors: JOHNSON E. , GAUPERAA T. , ESKELAND T.
Published: June 2006

We have studied the binding of fibronectin to complement (C3b, C3bi, C3d)‐coated agarose beads and its effect on cell association of such beads to mouse macrophages. Fibronectin bound to agarose beads preincubated in human serum, whereas no binding occurred after preincubation of the beads with complement‐inactivated (50°C for 20 min or ethylenediaminetetraacetic acid) sera. The binding of iodine‐labelled fibronectin to beads preincubated in fibronectin‐depleted serum (HS‐FIB) was about twice that of beads preincubated in normal serum. Unlabelled fibronectin inhibited the following binding of labelled fibronectin to beads pretreated in HS‐FIB. A similar amount of fibronectin bound to agarose beads coated with equimolar amounts of C3b, C3bi, or C3d, suggesting that the common domain C3d carries the main binding site(s) for fibronectin. Preincubation of serum‐treated and trypsinized agarose beads with fibronectin led to an increased association (22%) of such beads to mouse macrophages. The results indicate that fibronectin promotes binding of complement‐coated agarose beads to mouse macrophages, whereas the ingestion of the beads is mediated via complement C3 receptors.
Functional Analysis of Pokeweed Mitogen‐Dependent Cell Interactions in Murine Spleen Cells
Authors: BANDEIRA A. , POBOR G. , GULLBERG M. , COUTINHO A.
Published: June 2006

The nature of lymphocyte responses on addition of pokeweed mitogen (PWM) 10 normal murine spleen cells was studied in low cell density cultures. PWM, over a wide range of concentrations, stimulated proliferation in a set of cells roughly 10‐fold smaller than The lymphocyte populations responding to either concanavalin A or lipopolysaccharide. PWM also induced a relatively small number of B lymphocytes in these cultures to mature to Ig‐secreting plaque‐forming cells (PFC). Proliferative and PFC responses were completely abrogated by T‐cell removal from normal spleen cell cultures. Moreover, cell mixture and irradiation experiments demonstrated that B lymphocytes do not proliferate in response to PWM. even in the presence of an excess of normal T cells, suggesting that PFC development results from terminal maturation without proliferation. Finally, parallel titrations of cloned helper cells, normal splenic T cells or T‐cell blasts induced by PWM showed that the poor B‐lymphocyte responses in normal spleen cell cultures is due to the very low frequency of competen: helper cells in these populations. PWM, however, was competent to activate and expand this set of helper lymphocytes in primary cultures.
A Human Monoclonal IgG1λ Anti‐Hepatitis B Surface Antibody
Authors: STRICKER E. A. M. , TIEBOUT R. F. , LELIE P. N. , ZEIJLEMAKER W. P.
Published: June 2006

Peripheral blood mononuclear cells from a donor with a high litre of anti‐hepatitis B surface (HBs) antibodies were fused with a cell line that was positive for Epstein‐Barr virus nuclear antigen and sensitive to hypoxanthine‐aminopterine‐thymidine. A cell line was established that produces a monoclonal IgG1λ anti‐HBs antibody. Afterwards, it appeared that the anti‐HBs antibody‐producing cell line had arisen from Epstein‐Barr virus transformation of the donor B cells. The cell line is capable of producing up to 60 μg/ml of the monoclonal antibody, which has a high avidity for HBs antigen (Ag) and recognizes both ad and ay subtypes. The antibody is useful as a reagent for the detection of HBsAg in human serum. Over 1000 patient sera have been tested with a conventional third‐generation assay in parallel, and only a single discrepant serum was found.
The Effects of Topical Treatment with Steroids or Dithranol on Epidermal T Lymphocytes and Dendritic Cells in Psoriasis
Authors: BAKER B. S. , SWAIN A. F. , GRIFFITHS C. E. M. , LEONARD J. N. , FRY L. , VALDIMARSSON H.
Published: June 2006

We describe the effects of treatment with topical steroids or dithranol on T and dendritic ceils in the skin of patients with chronic plaque psoriasis. Resolution of lesions by both types of topical treatments was accompanied by a marked decrease in epidermal T cells. In steroid treated lesions there was also a reduction in DR+ dendritic cells to normal numbers during treatment and the rate of disappearance of both cell types correlated with the rate of resolution. However, a significant reduction of dendritic cells was not usually observed until after the T cells, had almost disappeared from the epidermis and substantial healing of lesions had taken place. Dendritic cells in steroid‐treated uninvolved skin had decreased to a lower level than in normal skin. In contrast, dithranol did not affect dendritic cells, either in lesional or in unaffected psoriatic epidermis. The decrease in dermal T cells observed with both treatments was more marked in steroid‐treated lesions and correlated with resolution. However, blood T cells were unaffected by both treatments. The findings provide further support for the role of T cells in the pathogenesis of psoriasis.
A Combination of Calcium Ionophore and 12‐O‐Tetradecanoyl‐Phorbol‐13‐Acetate (TPA) Stimulates the Growth of Purified Resting B Cells
Authors: GUY G. R. , BUNCE C. M. , GORDON J. , MICHELL R. H. , BROWN G.
Published: June 2006

In this study we investigated whether the calcium ionophores A23187 and ionomycin can act synergislically with the phorbol ester 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) to stimulate the growth of resting B lymphocytes purified from human tonsil cells, Ionomycin, A23187, and TPA added separately to cultures at doses of 0.4–1.6 μg/ml, 0.2–0.8 μ/ml, and 0.05–0.25 ng/ml respectively, did not induce DNA synthesis in resting B lymphocytes. In contrast, calcium ionophores at concentrations of 0.4–1.6 μg/ml ionomycin and 0.2–0.8 μg/ml A23187, in the presence of 0.05–4 ng/ml TPA, induced marked DNA synthesis and B‐cell proliferation, as shown by analyses of incorporation of [3H]thymidine, growth kinetics, and the percentage of cells in the S and G2+ M phases of the cell cycle. These results show that the synergistic effects of calcium ionophores and TPA can bypass the requirement for antigen and exogenous growth factors in B‐cell activation. These observations are similar to those obtained from studies of T lymphocytes by other workers.
Identification of a Cell‐Surface Glycoprotein Mediating Adhesion in Human Granulocytes
Authors: PATARROYO M. , BEATTY P. G. , SERHAN C. N. , GAHMBERG C. G.
Published: June 2006

Previous studies have shown that monoclonal antibody 60.3 reacting with a surface antigen common to human leukocytes inhibits phorbol ester‐induced adhesion among blood mononuclear cells and precipitates from these cells three surface polypeptides with apparent molecular weights of 90,000, 130,000 and 160,000. Now we report that the same antibody, either as purified IgG or Fab fragments, also inhibits the extensive adhesion among granulocytes induced by phorbol ester. Inhibition of cell aggregation was not observed with monoclonal antibodies to C3b receptor, common leukocyte antigen T200, C3bi receptor, brain granulocyte‐T lymphocyte antigen. IgG Fc receptor, class I transplantation antigen, or a granulocyte‐specific antigen. Intercellular adhesion induced by either the chemotactic tripeptide N‐formylmethionyl‐leucyl‐phenylalanine (FMLP) or the ionophore A23187 was also inhibited by antibody 60.3. However, this antibody did not affect phorbol ester‐induced superoxide (O2) generation or lysozyme release. Two major surface glycopolypeptides with apparent molecular weights of 92,000 and 155,000 were immunoprecipitated from granulocytes. Dissociation of the protein complexes obtained from blood mononuclear cells and granulocytes indicated the presence of the epitope on the 90,000‐92,000 molecular‐weight components. It is thus concluded that the smallest glycopolypeptides mediate adhesion in human granulocytes and mononuclear leukocytes.