bluuebunny/crossref_metadata_2025 huggingface dataset:
Selective Effects of Alpha Interferon on Human T‐Lymphocyte Subsets during Mixed Lymphocyte Cultures
Authors: HOKLAND M. , HOKLAND P. , HERON I. , SCHLOSSMAN S. F.
Published: June 2006

Mixed lymphocyte reaction (MLR) cultures of human lymphocyte subsets with or without the addition of physiological doses of human alpha interferon (IFN‐α) were compared with respect to surface marker phenotypes and proliferative capacities of the responder cells. A selective depression on the T4 (inducer) I ‐cell subset could be demonstrated as a sequence of events: decreased fluorescence intensity of the T4 inducer cells (day 2 of culture), decreased percentages of T4 cells as demonstrated by cell cytofluorometry (days 3–6 of culture), and decreased 3H‐thymidine incorporation of purified T4 cells and decreased numbers of T4 ceils harvested from IFN MLRs (days 5–6 of culture). In contrast, it was shown that the T8 (cytotoxic/suppressor) subset in MLRs was either not affected or slightly stimulated by the addition of IFN. The depression of the T4 cells by IFN was accompanied by a decrease in the number of activated T cells expressing Ia antigens. On the other hand, IFN MLRs contained greater numbers of cells expressing the T10 differentiation antigen. In experiments with purified T‐cell subsets the IFN effect was exerted directly on the T4 cells and not mediated by either T8 suppressor cells or monocytes. These findings are discussed in relation to other immunoregulatory effects of IFN‐α.
IgA Antibodies in Rat Bile Are Not Solely Derived from Thoracic Duct Lymph
Authors: DAHLGREN U. , AHLSTEDT S. , ANDERSSON T. , HEDMANJ L. , HANSON L. Å.
Published: June 2006

The IgA level in rat bile was significantly decreased by drainage of the thoracic duct, and passively administered IgA antibodies to Escherichia coli O antigen decreased similarity. In contrast, specific IgA antibodies against E. coli O antigen raised by immunization in the Peyer's patches did not diminish significantly in the bile. Rats immunized in the Peyer's patches with sheep erythrocytes had IgA‐forming cells in the thoracic lymph nodes, in the mesenteric lymph nodes, and in the spleen. Perfusion of the liver of immunized animals significantly decreased the bile levels of the IgA antibodies. It seems that IgA antibodies reach the bile not only via the thoracic duct but also via lymph ducts originating from thoracic lymph glands and the spleen.
Lymphocyte Populations in the Cerebrospinal Fluid and Peripheral Blood of Patients with Multiple Sclerosis and Optic Neuritis
Authors: SANDBERG‐WOLLHEIM M.
Published: June 2006

The cerebrospinal fluid (CSF) and peripheral blood (PB) of patients with multiple sclerosis (MS), optic neuritis (ON), and aseptic meningitis (AM) were studied with respect to the percentage of B cells (using membrane Ig fluorescence), T cells, and T‐cell subpopulations (using a rosetting technique or monoclonal antibodies). In the PB of all three patient groups there were normal B‐cell levels but a significant decrease in T cells compared with PB of normal individuals. In the CSF the B cells were reduced but the T cells elevated when compared with the PB of the patients, and these differences were statistically significant. The level of total T cells was not influenced by disease activity in MS or ON, but there was a significant reduction of suppressor cells in PB during disease activity in MS patients. In CSF there were also fewer suppressor cells during active disease, but the reduction was not statistically significant. Differences in B and T cells in CSF and PB indicate that the immune reactions within the central nervous system are at least partially isolated from the rest of the immune system. The changes in the T‐cell subpopulations in MS support the evidence for an immunoregulatory defect in this disease.
Soluble Cytostatic Factor(s) Released from Human Monocytes
Authors: EGGEN B. M. , BAKKE O. , HAMMERSTRØ J.
Published: June 2006

Human monocytes release a stable cytostatic factor during in vitro culture after stimulation with lymphokine and endotoxin. The cell cycle time of synchronized NHTK 3025 cells increased from 20.3 to 23.2 h in the first cell cycle when the target cells were exposed to the factor during the whole cell cycle. In exponentially growing NHIK 3025 cell cultures the cell doubling time increased from 18.9 h to 23. l h under continuous factor exposure for 70 h. These cells regained normal cell division rate when fresh culture medium replaced the cytostatic factor. Continuous exposure to the cytostatic factor for 96 h increased the cell doubling time of asynchronous K‐562 cells from 19.7 to 31.8 h. The target cell DNA synthesis, evaluated by thymidine incorporation. was markedly depressed after culture with the factor for 24 h, and this depression was detectable already within 4 h of culture. The factor showed no cytolytic effect on the two cell lines tested. The cytostatic factor influenced the cell cycle distribution of both target cells tested, since cell cycle analysis by DNA flow cytometry demonstrated a reversible inhibition of factor‐exposed NHIK 3025 cells in G1‐ and early S‐phase, whereas K‐562 cells accumulated in G1‐phase.
Cellular Responses to Bacterial Lipopolysaccharide: LPS Facilitates Priming of Antigen‐Reactive T Cells
Authors: MILNER E.C.B. , RUDBACH J.A. , VONESCHEN K.B.
Published: June 2006

The effect of bacterial lipopolysaccharide (LPS) on the capacity of mice to mount a specific T‐cell response to a protein antigen was examined. Inoculation of mice with LPS and ovalbumin (OA) resulted in enhancement of the T‐cell proliferative response to OA. This enhancement was manifested in vitro by an increase in magnitude and by a more rapid appearance of the response after challenge with OA. This enhancement was also shown because the latent periods for the antigen‐specific responses were reduced to 3 days after inoculation with antigen plus LPS, as compared with 5 days after inoculation with antigen alone. Various mouse strains, including the C3H/HeJ and C57BL/10ScN strains, responded to the adjuvant action of LPS at the T‐cell level. Results suggest that LPS exerts this adjuvant effect by facilitating clonal expansion of antigen‐reactive T cells during priming
T‐Lymphocyte Subsets and HLA‐DR‐Expressing Cells in Rejected Human Kidney Grafts
Authors: TUFVESON G. , FORSUM U. , CLAESSON K. , KLARESKOG L. , LARSSON E. , KARLSSON‐PARRA A. , FRÖDIN L.
Published: June 2006

This communication describes an immunohistochemical analysis of rejected human renal allografts. T‐lymphocyte subsets were identified in frozen tissue sections, by Leu 1 (anti‐ ‘pan’ T lymphocytes). Leu 2a (anti‐ ‘cytotoxic/suppressor’ T cells), and Leu 3a (anti‐ ‘helper/inducer’ T cells) monoclonal antibodies. In addition, HLA‐DR‐positive cells were identified by simultaneous labelling with heterologous anti‐HLA‐DR antibodies. T cells dominated the cellular infiltrates in acute cellular rejection. Leu‐3a‐positive cells were more numerous than Leu‐2a‐positive cells. The Leu‐3a‐positive cells usually appeared in clusters, whereas the Leu‐2a‐positive cells appeared scattered in the tissue. HLA‐DR‐positive non‐T cells were found within clusters of T ‘helper/inducer’ cells. The cell pattern shares many features with the findings in detayed‐type hypersensitivity reactions.
Mononuclear Cell Factors that Inhibit Fibroblast Collagen Synthesis
Authors: HART P. H. , POWELL L. W. , COOKSLEY W. G. E. , HALLIDAY J. W.
Published: June 2006

Peripheral blood mononuclear cells (PBMC) incubated either with phytohaemagglutinin‐P (PHA) under serum‐free conditions or with tuberculin purified protein derivative in the presence of 5% human blood group AB serum released into the culture medium factors that decreased collagen synthesis by dermal fibroblasts. These factors were first detected after incubation of PBMC with PHA for 24 h and were produced by non‐adherent cells. The decrease in collagen synthesis was observed despite changes in conditions of fibroblast culture, the addition of a cross‐linking inhibitor, and the use of different fibroblast cell lines.
T‐Lymphocyte Activation by a Gluten Fraction, Glyc‐Gli
Authors: SCOTT H. , FAUSA O. , THORSBY E.
Published: June 2006

A gluten fraction (glyc‐gli) induced proliferation of T lymphocytes obtained from coeliac patients or healthy controls. Glyc‐gli acted as an antigen, and T‐cell activation was strongly dependent on antigen‐presenting monocytes in the cultures. The lymphocyte response was similar for HLA‐B8/DR3‐positive and HLA‐B8/DR3‐negative healthy controls, and coeliac patients in remission on a gluten‐free diet could not be distinguished from the controls by the degree of proliferative activity in this test. Conversely, untreated coeliac patients showed a significantly lower lymphocyte response to glyc‐gli than both coeliac patients in remission and controls. This result may reflect sequestration of gluten‐specific cells into the mucosal lesion or mesenteric lymph nodes in patients with active coeliac disease.
Complement (C3)‐Receptor‐Mediated Phagocytosis of Agarose Beads by Mouse Macrophages
Authors: JOHNSON E. , ESKELAND T.
Published: June 2006

The phagocytosis by macrophages of C3bi‐coated agarose heads reached a plateau after 15 min, compared with 30 min for C3b‐coated beads. By using 125I‐Iabelled C3bi or C3b coupled to the agarose beads, we found that 70% and 95% of total radioactivity were removed from the heads after 12 h and 36 h of intracellular digestion, respectively. Intracellular degradation of C3bi linked to agarose beads was also demonstrated by testing binding of monoclonal antibodies against human C3c, C3g and C3d to beads extracted from the cells after phagocytosis. Such extracted beads also showed reduced attachment to new macrophages compared with non‐ingested beads. Treatment of the cells with leupeptin, an inhibitor of the lysosomal enzyme cathepsin B, or with dextran sulphate to inhibit phagosome‐lysosome fusion greatly reduced the release of labelled protein from the agarose during the first 12 h. These findings show that C3bi and C3b on agarose is destroyed intracellularly by lysosomal enzymes.
Characterization and Functional Studies of the Murine T‐Lymphocyte Response to Mycobacterium leprae Antigen
Authors: HAREGEWOIN A. , LOUIS J.
Published: June 2006

Mice were immunized with, Mycobacterium leprae in incomplete Freund's adjuvant, and sensitized lymphocytes were obtained from draining lymph nodes. The lymphocytes thus obtained proliferated specifically in vitro in the presence of M. leprae antigen, and this response was shown to be both T‐ccll and macrophage dependent. T‐cell blasts generated in vitro in response to M. leprae antigen were grown in the presence of interlcukin‐2 (IL‐2), The proliferative response of these blasts to M. leprae antigen was strictly dependent on the presence of syngeneic spleen cells as antigen‐presenting cells. M. leprae‐Immune F, blasts responding to the antigen in the context of either parental H‐2 haplotype‐hearing accessory cell could be obtained by positive selection from an F, hybrid‐responding cell population. By means of flow microfluorometry the T‐cell phenotype of the M. leprae‐specific T‐cell blasts was found to be Thy‐1+ and to be composed of Lyt‐1+ and Lyt‐2+ subpopulations. Functionally, the blasts were shown to transfer delaycd‐lype hypersensitivity locally to non‐immunized recipients and to have cytolytic activity. Limiting dilution analysis showed the frequency of M. leprae‐responding cells from blasts grown in IL‐2 to be approximately 1/333.
Kinetic Analysis of the Specificity of Human Natural Cytotoxicity
Authors: CALLEWAERT D. M. , MAHLE N. H. , DAYNER S. , EARNSHAW B. , BOWMAN S. , O'CONNOR M.
Published: June 2006

Distribution‐free analysis of kinetic data for cellular cytotoxicity reactions enables the precise determination of kinetic parameters for the lysis of target cells by individual lymphocyte preparations. This report presents results obtained when this method was used to quantitate the inhibition of human natural cytotoxicity by unlabelled (cold) target cells. In agreement with previous studies, we found that the natural cytotoxicity of a given target cell can be inhibited by heterologous as well as homologous unlabclled cells: however, the strongest inhibition was usually produced when the unlabelled inhibitor cells were homologous to the labelled target cells. The general pattern of inhibition observed for the target and inhibitor cells tested in these experiments was competitive, and when inhibitor cells were homologous to the labelled cells, the observed increase in Kmapp agreed with theoretical predictions based en equations derived previously. These results support earlier reports of limited but not absolute antigenic specificity by subsets of human NK cells. Moreover, while Kmapp values for cytotoxicity reactions are not simply related to antigen binding, quantitative analysis of the relative inhibition of cytotoxicity by heterologous versus homologous unlabclled cells provides useful estimates of the relative affinity of the effector cell subset(s) that kill a given target cell for various NK target cells.
Spontaneous Cytotoxicity by Monocyte‐Enriched Subpopulations of Human Peripheral Blood Mononuclear Cells against Human or Mouse Anchorage‐Dependent Tumour Cell Lines
Authors: CHANG Z. L. , HOFFMAN T. , BONVINI E. , STEVENSON H. C. , HERBERMAN R. B.
Published: June 2006

Human peripheral blood mononuclear cells (PBMNC) were found to be cytotoxic for mouse or human anchorage‐dependent target cell lines in a 48‐72 h [125I)iododeoxyuridine (IUDR) release assay. Unfractionated, adherent or nonadherent cells had significant levels of cytotoxicity, as did cells fractionated according to size into ‘lymphocytes’ or ‘monocytes’ by elutriation. Intermediate size cells, not enriched for monocytes, had high levels of cytotoxicity. In all fractions tested, including adherent populations, some cells with the morphology of large granular cells were observed. Treatment of all fractions with interferon (IFLrA, a purified. recombinant α‐IFN) boosted cytotoxicity against four target cells lines. Treatment with lymphokines containing putative‘macrophage‐activating factor’(MAF) also enhanced cytotoxicity in fractions depleted of monocytes. Culture in fetal bovine serum enhanced cytotoxicity mainly in unfractionated and nonadherent PBMNC. These experiments indicated that N K‐like cells can be appreciable contaminants in clutriator‐purified monocyte‐enriched or adherent cell populations and thereby contribute to observed cytotoxicity, particularly after pretreatment with IFN or other stimulatory factors.
Effect of Human Monocyte‐Released Cytostatic Factors on Target Cell DNA Synthesis, Protein Synthesis, and ATP Content
Authors: NISSEN‐MEYER J. , SEIM S.
Published: June 2006

Cultured human monocytes activated in vitro with lymphokines and lipopolysacchuridc release cytostatic protein factors that may be separated by ion‐exchange chromatography into two populations, termed cytostatic factors I and II (CF I and II). The effect of CF I and II on target cell DNA synthesis, protein synthesis, and ATP content and the possible presence of a CF‐associated protease activity were investigated. Inhibition of DNA synthesis was detected 4 h after addition of either CF I or CF II, whereas inhibition of protein synthesis was first detected after 10‐15 h. Moreover, 20 h after addition of either CF I or CF II the average DNA synthesis per cell was inhibited by 25–35%, as compared with 10–15% inhibition of protein synthesis. No significant alteration in cellular ATP content was observed in cells culture, up to 25 h with either CF I or CF II. Thus, neither protein synthesis nor generation of ATP appeared to be primary targets of either CF I or CFII, but the factors may act more directly on DNA synthesis. Protease activity was not associated with either CF I or CF II, and the factors thus differed from a previously reported cytolytic protease released from activated murinc macrophages.
Residual Minor Histocompatibility Genes Contaminate the B10. AM Congenic Line: No Evidence of C‐Region‐Controlled Histoincompatibility
Authors: JURETIĆ A. , PROTRKA N. , WALDEN P. , NAGY Z. A. , KLEIN J.
Published: June 2006

After being primed in vivo and restimulated in vitro, cytolytic T lymphocytes (CTL) were produced in the strain combinations B10.AM ami‐B10.A(1R) and B10.A(1R) anti‐B10.AM. Although the two strains differ in the chromosomal interval between the Eα and the D loci, the CTL are not directed against antigens controlled by loci in this interval. Instead, the CTL detect minor histocompatibility (H) antigens controlled by loci that are notlinked to H‐2. The recognition of the antigens detected by the B10. AM anti‐B10.A(lR) CTL is restricted by the Kk and Db molecules, but the CTL also cross‐react with the Dd molecule (or a molecule controlled by a locus closely linked to Dd). The recognition of the antigens detected by these two CTL behave as if controlled by alleles at the same minor H locus or loci. This locus is distinct from H‐2, and the B 10. AM congenic line apparently retained a C3H‐derived allele at this locus.
Functional Differences of Anti‐T‐Cell Antibody in Patients with Systemic Lupus Erythematosus and Ulcerative Colitis
Authors: ABE T. , MORIMOTO C. , TOGUCHI T. , KIYOTAKI M. , TAKEUCHI T. , KOIDE J. , ASAKURA H. , TSUCHIYA M. , HOMMA M.
Published: June 2006

The loss of suppressor T‐cell function results in an abundant production of autoantibodies in systemic lupus erythematosus (SLE). As a cause of this suppressor T‐cell defect, anti‐T‐cell antibody seems to be of prime importance. On the other hand. anti‐T‐cell antibodies can be detected in various other autoimmune diseases, but their functional characteristics have not been determined. In the present study, the functional characteristics of anti‐T‐cell antibody from a selected subgroup of patients with ulcerative colitis (UC) were compared with those from patients with SLE. Anti‐T‐cell antibody from the patients with SLE reacted with a T8 subset, resulting in a suppressor defect, whereas anti‐T‐cell antibody from the UC patients reacted primarily with a T4 subset. Functionally, SLE T cells failed to proliferate in response to concanavalin A. whereas UC− T cells from UC patients failed to proliferate in response to phytohaemagglutinin. In the Ig synthesis system, both SLE− and UC− T cells increased Ig production of B cells. Since UC+ T cells did not contribute to the generation of Con‐A‐inducible suppressor activity, we believe that serum from the selected subgroup of patients with UC reacted with the inducer T‐cell subset.
Increased Immunoglobulin Production in Vitro in Healthy HLA‐B8‐Positive Persons
Authors: VUENTO R. , ESKOLA J. , LEINO R. , KOSKIMIES S. , VIANDER M.
Published: June 2006

The histocompatibility antigens B8 and DR3 are associated with several autoimmune diseases, and immune hyper‐responsiveness has been believed to be one factor in the pathogenesis of these diseases. In the present study we compared the in vitro immunoglobulin production of healthy HLA‐B8‐positive individuals with that of HLA‐B8‐negative individuals. In HLA‐B8‐positive persons a higher number of pokeweed mitogen (PWM)‐stimulated Ig‐secreting cells was obtained. When the cells were stimulated with PWM in the presence of a physiological concentration of hydrocortisone (HC), the secretion of IgG was significantly higher in HLA‐B8‐positive individuals. This finding indicates that the PWM‐stimulated IgG secretion in healthy HLA‐B8‐positive individuals is more sensitive for the enhancing action of HC.
Amino Acid Sequences in Amyloid Proteins of χIII Immunoglobulin Light‐Chain Origin
Authors: SLETTEN K. , WESTERMARK P. , PITKÄNEN P. , THYRESSON N. , OLSTAD O. K.
Published: June 2006

The main amyloid fibril (AL) proteins extracted from the spleen of Patient So 124 with systemic amyloidosis and from a skin nodule of Patient KSA with localized amyloidosis were studied by partial amino acid sequence analysis and proved to be of χIII immunoglobulin light‐chain origin. The sequences were similar to that of Bence Jones protein Vand, which has been reported to have a unique χIII subset sequence. Thus, except for position 9 in protein AL(KSA), the amino acid sequences were identical to position 25 in AL(So 124) and in AL(KSA). The question is being raised whether this χIII subset might contain amyloidogenic sequences.
Studies on the Role of Mononuclear Phagocytes in Resistance to Acute Lymphocytic Choriomeningitis Virus Infection
Authors: THOMSEN A. RANDRUP , VOLKERT M.
Published: June 2006

The role of mononuclear phagocytes in various phases of the acute lymphocytic Choriomeningitis virus (LCMV) infection was studied. The anti‐macrophage agent carrageenan delayed virus clearance. Carrageenan was most effective when given before virus inoculation, suggesting that it interfered with early events in the host response to the virus. Correspondingly, carrageenan enhanced early virus multiplication. Pretreatment with carrageenan apparently did not inhibit induction of the T‐cell response and had little or no direct effect on T‐cell‐dependent anti‐viral activity. The LCMV‐induced natural killer response was also unimpaired by this treatment. Taken together, these findings suggest that resident macrophages constitute a barrier to the initial multiplication of LCMV. A breakdown of this macrophage barrier results in a more disseminated infection, which the specific immune response has difficulty in eliminating. Adoptive transfer experiments with pre‐irradiated recipients showed that T‐cell‐dependent virus clearance required interaction between donor‐derived primary immune spleen cells and radiosensitive host cells. T cells did not seem to constitute the radiosensitive host component, since athymic (nude) mice functioned well as recipients. Together with previously published data, this finding strongly suggests that T‐cell‐dependent virus clearance involves cooperation between T cells and non‐committed cells, probably monocytes.
Class‐II HLA Restriction of Antigen‐Specific Human T‐Lymphocyte Clones
Authors: QVIGSTAD E. , THORSBY E.
Published: June 2006

Blast‐enriched suspensions of T cells primed for Chlamydia trachomatis antigen were cloned by a limiting dilution technique. The class‐II HLA restriction of T‐lymphocyte clones (TLC) was studied by using allogeneic antigen‐presenting cells (APC) carrying foreign class‐II HLA antigens. Most of the TLC were restricted by one or the other of the D/DR determinants of the T‐cell donor; that is, they did not respond when antigen was presented by APC expressing foreign D/DR determinants. Furthermore, heterogeneity of the DR4‐expressing molecule could be demonstrated by T‐cell clones from one person; APC from family members expressing DR4 gave high proliferative responses, whereas no proliferation was observed with most APC from unrelated persons expressing DR4. This heterogeneity of DR4 was confirmed by mixed lymphocyte culture (MLC) experiments, indicating a close relationship between restriction epitopes and those that activate allogeneic T cells. Other clones seemed to be restricted by other class‐II HLA determinants, most probably MT determinants of the T‐cell donor. The restriction specificities were confirmed by subcloning experiments.
T‐Cell Growth Factor Contains Activity that Supports Natural Killer Activity in Vitro
Authors: GRÖNVIK K.‐O. , ANDERSSON J. , ALM G. , STERN P.
Published: June 2006

Lymphocytes from athymic and normal mice were tested and compared for cytotoxic activity after in vitro cultures suppemented with concanavalin A or T‐cell growth factor (TCGF). Our results indicate that, in addition to cytoloxic T cells, natural killer (NK) activity can be recovered from lymphocytes cultured in the presence of TCGF and that this is not the result of contaminating interferon in the TCGF preparations. The NK nature of the effector cells was established by means of a panel of target cells, including a teratocarcinoma tumour cell, which enabled the distinction of T‐cell‐ and NK‐cell‐mediated lysis.
T‐Cell Immunoregulatory Functions in Rheumatoid Arthritis Patients
Authors: EGELAND T. , LEA T. , MELLBYE O. J.
Published: June 2006

We have studied the immunoregulatory function of T8+ (suppressor/cytotoxic) and Leu3a+ (inducer/helper) T cells from rheumatoid arthritis (RA) patients by measuring the effect of these T‐cell subpopulations on the generation of immunoglobulin‐secreting cells by normal allogeneic B cells after stimulation with pokeweed mitogen (PWM) in vitro. When T8+ or Leu3a+ cells from blood or synovial tissue from nine patients were substituted for T8+ or Leu3a+ cells, respectively, from normal blood mononuclear cells (MNC). RA T8+ cells showed an increased suppressor activity, whereas RA Leu3a+ cells were, except for one patient, weak augmentors. Unreplaced normal MNC and MNC replaced with allogeneic normal T‐cell subpopulations responded equally to PWM. When T8+ plus Leu3a+ cells from the same patient replaced normal T cells, high B‐cell responses were detected. Normal T8+ plus Leu3a+ cells generally supported the response to a lower degree. Substitution with two allogeneic T‐cell subpopulations did not result in a B‐cell response to PWM. Thus, whereas RA T8+ seemed to be strong suppressors and RA Leu3a+ cells weak augmentors by themselves, together they are possibly able to generate a B‐cell stimulatory potential that might be of pathogenetic significance in the patients.
A Common Epitope Identified by a Monoclonal Antibody, MID 2, Present on All Leucocytes and Associated with a Group of High Molecular Weight Glycopeptides
Authors: BANGA J. P. , GUARNOTTA G. , HARTE A. , PRYCE G. , CAMPBELL M. A. , QUARTEY‐PAPAFIO R. , LYDYARD P. M. , ROITT I. M.
Published: June 2006

A monoclonal antibody, MID 2, which reacts with an epitope common to all human leucocytes, was used to show that the leucocyte common antigen can be resolved into four glycoprotein components with molecular weights of 220 K. 200 K, 180 K and 160 K. The 200, 180 and 160 K peptides were all present to various extents in the T and B lymphoblastoid cell lines examined, but the 220 K component was only detected in the B‐cell lines. The 220 K glycoprotein was also lacking in human thymocytes, but evidence of its expression in peripheral blood T lymphocytes was obtained, suggesting that it might be acquired as the cells differentiated. The four glycoproteins isolated from tonsil cells gave similar patterns on peptide mapping by the Cleveland enzymatic method or by cyanogen bromide cleavage, indicating extensive homology. It is conceivable that they differ from each other by the acquisition of simitar repeating domain sequences of approximately 20 K. Alternatively, they may share a common peptide structure but differ in their electrophoretic behaviour in SDS gels owing to differences in carbohydrate; if this is the case, however, experiments with tunicamycin suggest that O‐linked oligosaccharides must be involved.
A Routine Source of Human Peritoneal Macrophages
Authors: MADDOX Y. , FOEGH M. , ZELIGS B. , ZMUDKA M. , BELLANTI J. , RAMWELL P.
Published: June 2006

The routine availability of nucleated human cells for experimental use is limited in the absence of venipuncture. In this paper we have demonstrated that macrophages may he harvested routinely from the waste dialysis hags of patients undergoing continuous ambulatory peritoneal dialysis. These cells were identified as macrophages by morphology, adherence, phagocytosis, chemotaxis, non‐specific esterase staining and peroxidase staining. Macrophages from patients with end‐stage renal disease produced arachidonate cyclo‐oxygenase products in a pattern similar to that of ascitcs macrophages obtained from patients with normal kidney function. Arachidonate metabolism was shown to be manipulatable. Thus, indomethacin blocked synthesis of cyclooxygenase products, and OKY‐1581, a specific thromboxane synthase inhibitor, increased the release of prostagtandin E2 and prostacyclin, measured as its stable breakdown product 6‐keto‐prostaglandin F1, whereas the thromboxane B2 synthesis was effectively inhibited.
The Chemotactic Activity of Normal and Complement‐Activated Serum as Measured by the Leading‐Front Method Using a Boyden Chamber
Authors: HÅKANSSON L. , VENGE P.
Published: June 2006

The chemotactic activity in serum, defined as the attractant effect of serum on the migration of neutrophil granulocytes (PMN) has been investigated for the purpose of characterizing the major chemotactic factors in serum as measured by the leading‐from technique, using a modified Boyden chamber. The chemotactic activity was measured in fresh and heated normal and activated serum and in serum fractions thereof separated by gel filtration. By gel filtration on Sephacryl S‐200 a partly heat‐labile C3‐C5‐associated chemotactic factor with molecular weight between 70,000 and 150,000 was isolated from fresh normal serum. The heat‐labile chemotactic activity was destroyed by pronounced complement activation. Gel filtration of complement‐activated serum on a Sephacryl S‐200 column showed the existence of one C5‐associated chemotactic factor with ˜70,000 molecular weight and one unidentified factor with ˜ 150.000 molecular weight, whereas no low molecular weight chemotactic activity was demonstrated. On the other hand, get filtration of activated serum on a Sephadex G‐75 column demonstrated one C5‐associaied chemotactic factor of ˜ 70,000 molecular weight and one 10,000–50,000 molecular weight factor active only in the presence of 2% normal serum. This investigation suggests that the chemotactic activity in fresh normal serum is mediated by a partly heat‐labile C3–C5‐associated complex. In activated serum three chemotactic factors were demonstrated, one unidentified factor with 150,000 mol wt and two CS‐dependcnt factors with 70,000 and 10,000–50,000 mol wt, the latter probably corresponding to C5adesarg. Accordingly, this study also suggests that C5a is not the only chemotactic factor generated in serum.
Human Keratinocytes Express HLA‐DR Antigens in the Tuberculin Reaction
Authors: SCHEYNIUS A. , TJERNLUND U.
Published: June 2006

The cellular response in the human skin tuberculin reaction was studied with immunohistochemical double‐staining techniques in frozen sections of skin biopsies taken 6 h to 8 days after intradermal PPD injections. Cell infiltrates were observed from day 2 onwards and increased in size up to 4 days. Most of the infiltrating cells reacted with anti‐Leu 3a (T ‘helper/inducer’ phenotype) antibodies. In contrast to normal epidermis, not only Langerhans cells but also keratinocytes expressed MLA‐DR antigens from day 4 onwards. The induction of HLA‐DR antigens on keratinocytes may be secondary to T‐cell activation. Since the HLA‐DR expression on keratinocytes appeared late in the tuberculin reaction, the function may be to suppress rather than enhance the immune response.
Decreased Cloning of Lymphocytes from Elderly Individuals
Authors: CHRYSOSTOMOU A. , SESHADRI R. , MORLEY A. A.
Published: June 2006

Limiting‐dilution cloning of human lymphocytes was used to study cloning and proliferation of lymphocytes in 10 young and 10 elderly individuals. The cloning efficiency for elderly individuals, 3.5%, was significantly lower than that for young individuals, 19.8%. Mixing experiments showed that this decrease was due to both a decrease in the frequency of the clone‐forming cells and a decrease in the interacting ability of the lymphocytes. Lymphocytes from elderly individuals which did form clones, however, proliferated at the same rate as did those from young individuals.
Functional Heterogeneity of Human Cord Blood Monocytes
Authors: KHANSARI N. , FUDENBERG H. H.
Published: June 2006

Human cord blood monocytes were separated into four different subpopulations by means of a discontinuous bovine serum albumin (BSA) gradient. Of the least dense, 31% were present in fraction A, 17% in fraction B and 13% in fraction C and of the most dense, 20% were in fraction D. The rest (17%) sedimented as a pellet, of which 93% were dead cells. The monocytes of fraction C (density ≤ 1.070) demonstrated suppressor activity on in vitro antibody synthesis of maternal B cells. Fraction D (density ≤ 1.075) monocytes enhanced antibody synthesis of maternal B cells compared with synthesis produced in a similar experiment with unfractionated monocytes. Addition of either fraction A or B monocytes to the mixed culture of T and B cells resulted in antibody production comparable to that produced by addition of unfractionated monocytes. The functional heterogeneity of the cord monocytes was assayed also by 2‐deoxyglucose (2‐DOG) uptake, a marker for immunological macrophage activation. Fractions A and D showed significantly higher 2‐DOG transport than that of unfractionated monocytes; in contrast, fraction C showed a 50% reduction of 2‐DOG uptake. Furthermore, in contrast to fraction D, fraction C possessed only minimal phagocytic activity (for antibody‐coated sheep erythrocytes) and minimal hemoxygenase enzyme activity. These data demonstrate functional heterogeneity of cord blood monocytes.
T‐Cell Growth Factor Production and Protein Synthesis
Authors: VOITENOK N. N. , VARIVOTSKAYA N. V.
Published: June 2006

It is known that protein synthesis inhibitors (PSI) do not block production of some immunoregulatory and inflammatory mediators. Here we show that the production of T‐cell growth factor (TCGF) by phytochaemagglutinin‐stimulated human peripheral blood lymphocytes (PBL) was resistant to inhibition of protein synthesis more than 90–95%. Moreover, TCGF production was significantly accentuated in the presence of low concentrations of PSI in cultures from low‐ and high‐producer donors. The presence of macrophages was non‐essential for enhancement of TCGF production by PSI. When high doses of PSI were used, TCGF production was inhibited. In contrast, the synthesis of granulocyte/macrophage colony‐stimulating factor (CSF) was blocked in the presence of low concentration of PSI. Hence, the results indicate that human TCGF produced by PBL belongs to a group of soluble factors whose production is relatively resistant to the action of PSI.
Plaque‐Forming Cells in Man
Authors: TAURIS P. , JOHNSEN H. E. , CHRISTIANSEN S. E. , KISSMEYER‐NIELSEN F.
Published: June 2006

We have investigated the ability of allogeneic, irradiated T lymphocytes to induce proliferation and immunoglobulin (Ig) secretion in untreated peripheral blood B lymphocytes. Non‐mitogen‐activated co‐cultures of isolated T and B lymphocytes from selected, full‐house HLA‐A, B and D/DR antigen‐phenotyped donors were reconstituted in a ratio of 4:1. Proliferation was assessed on day 5–6 of culture by the 3H‐thymidine incorporation technique, and the Ig secretion was monitored on day 6 with a protein A plaque‐forming cell (PFC) assay. B lymphocytes were able to diffferentiate into PFC, and the number of plaques was significantly higher in cultures of cells with two HLA‐D/DR antigen incompatibilities than in those sharing one antigen. In cultures of peripheral blood lymphocytes with no HLA‐D/DR antigen difference, only a few PFC developed. HLA‐A and B antigens had no influence on the response. Further, monocytes were not an absolute requirement for allogeneic activation of B cells. Sonicated T cells and culture supernatants from allogeneic T‐ and B‐cells cultures were not able to induce PFC formation in B lymphocytes. Our results indicate that the PFC response obtained in non‐mitogen‐activated cultures of allogeneic T and B lymphocytes is dependent on HLA‐D/DR disparity or on genes encoded in the HLA‐D/DR region.
Enumeration andCharacterization of Human Killer and Natural Killer Cells by a Modified Single‐Cell Assay
Authors: WÅHLIN B. , ALSHEIKHLY A. , PERLMANN P. , SCHREIBER R. D. , MÜLLER‐EBERHARD H. J.
Published: June 2006

Human natural killer (NK) and killer (K) cells were assayed in a modified single‐cell cytotoxicity assay using poly‐L‐lysine‐coated cover slips. When human Chang liver cells were used as targets, 20% of the lymphocytes formed conjugates and 2% were active NK cells. When anti‐Chang antibodies were present, the proportion of target‐binding cells (TBC) increased to 30% and that of the cytotoxic effector cells (comprising NK + K) to 6%. With the mouse mastocytoma cells (P815), which are not susceptible to NK, similar proportions of lymphocytes formed conjugates, and 6–9% were active as K cells. By an in situ rosetting assay a significant fraction of the TBC and cytotoxic effector cells bound either C3b or C3bi in both systems, with a certain predominance of C3bi‐binding cells among the K cells. However, by indirect immunofluorescence, significantly more OKT3+ cells than OKM1+ cells were TBC or cytotoxic in the Chang cell system, whereas the OKT3+/OKM1+ ratios for both TBC and cytotoxic cells were 1:1 in the mouse mastocytoma system. The results indicate that TBC, NK and K cells are heterogeneous with respect to surface marker expression and that effector cells of different phenotypes predominate in different target systems.
Naturally Occurring Spleen‐Associated Suppressor Activity of the Newborn Mouse
Authors: JADUS M. R. , PECK A. B.
Published: June 2006

Spleens from newborn mice less than 6–7 days of age are known to contain naturally occurring suppressor cells, which can suppress the immune reactivity of third‐party adult cells. In the present study newborn spleen cell populations are shown to possess the potential to inhibit lethal graft‐versus‐host (GVH) disease in sublethally gamma‐irradiated hosts injected with allogeneic adult cells. However, this capacity to suppress GVH disease is controlled by at least two genetic restrictions: (1) the newborn spleen cells and the adult donor cells must be histocompatible at an H‐2‐linked region apparently telomeric of H‐2DL, and (2) the newborn spleen cells must express a strongly stimulating non‐H‐2 (perhaps Mis) alloantigenic phenotype. Host animals that survive GVH remain chimeric for at least 3–4 weeks but return to the host phenotype by 8–10 weeks. Thus, it appears that in sublethally irradiated hosts the newborn cells suppress donor cell reactivity long enough for the host system to recover from the effects of irradiation.
Polymorphonuclear Leucocytes Defective in Oxidative Metabolism Inhibit in Vitro Growth of Plasmodium falciparum
Authors: KHARAZMI A. , JEPSEN S. , VALERIUS N.H.
Published: June 2006

This report presents evidence that polymorphonuclear leucocytes (PMN) from chronic granulomatous disease (CGD) patients, who are defective in oxidative metabolism, are capable of inhibiting in vitro multiplication of Plasmodium falciparum. Using a microtitre in vitro inhibition assay, we incubated various numbers of peripheral blood neutrophils from CGD patients and from normal individuals with P. falciparum isolate F32 in the in vitro culture system. Inhibition of parasite growth by neutrophils was determined after 48 h of culture. At PMN to erythrocyte ratio of 1:50 there was an inhibition of parasite growth of 57% by normal neutrophils and 39% to 68% by CGD cells. When the neutrophils were stimulated by phorbol myristate acetate, both cell types enhanced inhibition of parasite growth. These findings indicate that the oxygen‐independent systems of human neutrophils are involved in parasite destruction. Constituents of neutrophil granules such as acid hydrolases, lactoferrin, and cationic proteins could be regarded as potential mediators of parasite destruction.
The Appearance of Cytotoxic Lymphocytes in Unstimulated and Concanavalin A‐Activated Human Peripheral Mononuclear Cells
Authors: PERSSON U. , JOHANSSON S. G. O.
Published: June 2006

The presence of a mitogenic lectin during the cytotoxicity assay enables the detection of non‐specific cytotoxicity (lectin‐dependent cell‐mediated cytotoxicity (LDCC). Human mononuclear cells cultured at 37°C in vitro became cytotoxic after 1 day, as detected by the LDCC. In some individuals, the presence of adherent cells during culture inhibited the development of cytotoxicity. Activation by concanavalin A (Con A) resulted in enhanced cytotoxicity after a few days in culture. There was no clear relationship between the lectin doses required for optimal induction of cytotoxicity and [3H]|thymidine uptake. High Con A concentrations that induced a proliferate response often inhibited cytotoxicity in mononuclear cells depleted ot adherent cells.
Effect of Maternal Antigenic Stimulation on the Active Immune Responses of Their Offspring
Authors: WATANABE Y. , SHIMIZU S. , YAMAGUCHI N.
Published: June 2006

Pregnant mice were stimulated by heterologous erythrocyte and protein antigen, and the active immune responses of their offspring as measured by plaque‐forming cells (PFC) were investigated. In offspring derived from mothers immunized by a suitable amount of T‐dependent antigen, clear‐cut suppression of development of specific PFC in spleen was observed over a significant period after delivery. The mechanism of this suppression was investigated, and the following results were obtained. When the heterologous erythrocyte was used as antigen, the more the specific PFC developed in the mother spleen, the stronger the suppression of PFC observed in their offspring. However, it is worthy of note that passive administration of antibody to pregnant mice did not induce suppression in their young. In case of the protein antigen ovalbumin (OVA), pregnant mice had to be injected with a suitable amount of antigen, along with aluminium hydroxide, for either primary or secondary stimulus to induce the suppression of specific PFC in their offspring. Soluble OVA administered to pregnant mice was not effective for inducing suppression in the offspring. Based on these results, some possible mechanisms are discussed concerning specific PFC suppression in the offspring when pregnant mice are stimulated.
Role of HLA Class II Products in Proliferative T‐Lymphocyte Responses to PPD
Authors: van EDEN W. , ELFERINK B. G. , HERMANS J. , de VRIES R. R. P. , van ROOD J.J.
Published: June 2006

HLA class II determinants were analysed for their role in monoeyte‐T‐cell infractions in the prolifeniuve response to purified protein derivative (PPD). Allogeneic cell combinations of monocytes and T cells were tested with a range of suboptimal PPD concentrations. For each cell combination tested, a summary measure to characterize the antigen‐induced response curve was calculated by means of regression analysis of the response in the dose of PPD added, thus obviating a further need to correct for mixed lymphocyte reactivity. In 200 distinct monocyte‐T‐cell combinations. HLA‐DR appeared io be the major restricting clement. Additionally, a marginal influence of HLA‐MB or ‐MT matching was observed and no evidence of HLA‐SB‐restricted responses. Remarkably, the HLA‐MB1 specificity was significantly associated with low responsiveness in DR sharing monocyte‐T‐cell combinations, indicating a modulating role of MB1 in DR‐restricted T‐cell responses.
Peritoneal B Cells Differentiate without Proliferation into Autoantibody Secretors under the Influence of Factors Released by Other Cells
Authors: COX K. O. , DAENKE S. , SAMCEWICZ B.
Published: June 2006

After 3–5 days of in vitro culture, peritoneal cells from untreated Oil mice produce autoantibodies specific for autoantigens in the membranes of mouse crythrocytes. The autoantigens are exposed in vitro by treating mouse erythrocytes with the proteolytic enzyme bromclain. Limiting‐dilution cell culture techniques were used to determine the frequency of the autoreactive B cells. Cells were cultured in Terasaki trays at 10–200 cells/well. Autoantibody production was assayed with an in situ plaque‐forming cell (RFC) assay. On average, one autoantibody precursor cell was detected for every 150–200 peritoneal cells cultured. This precursor frequency was increased to 1 in 50 by the addition of lipopolysaccharide (LPS) and dextran sulphate (DS) to the culture medium The addition of a culture supernatant from an EL‐4 lmphoma subline also induced a high proportion of the peritoneal cells to secrete autoantibodies However, it was not possible to determine thefrequency of PFC accurately because at limning numbers of peritoneal cells the effects of EL4 affected more than one limiting variable. Significant cell division was observed in cultures to which LPS/DS had been added, in contrast to untreated cultures to which EL4 supernatant was added. The results show that high numbers of autoreactive B cells committed to self antigens are present in the peritoneal cavity and that these cells under the influence of appropriate cytokines can differentiate in vitro, even without proliferation, into autoantibody secretors. The cell type or types releasing the cytokines remain to be identified.
Human Peripheral Blood Null Lymphocytes Stimulated by Staphylococcus aureus Cowan I Produce Atypical Acid‐Labile Interferon in Vitro
Authors: FUNA K. , RAMSTEDT U. , RÖNNBLOM L. , ALM G. V.
Published: June 2006

Peripheral blood mononuclear leucocytes (PBLs) stimulated in vitro by heat‐killed formaldehyde‐fixed Staphylococcus aureus Cowan I (SACoI) produced acid‐labile alpha interferon (IFN‐α) and to various extents, also IFN‐γ. The IFN producers resided in nylon wool‐nonadherent cells, and monocytes suppressed SACol‐induced IFN responses. Further separation of nonadherent PBLs in accordance with expression of surface antigenic markers was performed with a ‘panning’ technique. The SACoI induced production of IFN in cells that carried neither surface immunoglobulins nor OKT3‐defined antigens. These cells were also characterized as OKM1‐ and OKT10‐negative. In contrast, cells with natural killer (NK) activity against K562 erythroleukaemia cells were located in both OKM1‐ and OKT10‐positive and ‐negative cells. At centrifugation on Pereoll density gradients, cells with NK activity and IFN response against SACoI were recovered from light gradient fractions that contained mainly large granular lymphocytes (LGL). Furthermore, the IFN producers were enriched by removal of sheep erythrocyte‐rosetting T cells from the Percoll fractions. These SACoI‐indueed IFN‐producing PBLs are LGL, but lack certain antigens that are frequently found on NK cells.
Effect of Cyclosporin and Interleukin‐2 on the Restoration of in Vitro Immune Responses to Cytomegalovirus
Authors: CONVERSE P. J. , HESS A. D.
Published: June 2006

Previous studies have shown that cyclosporin (CsA) inhibits lymphoproliferation to Cytomegalovirus (CMV)‐infected, glutaraldehyde‐fixed, and irradiated fibroblasts (CMVFFX) in vitro. Generation of cytotoxic cell activity is impaired in cultures with CsA, but the induction of suppressor cells is not. In the present studies we tested the ability of interleukin‐2 (IL‐2) and supernatants of lymphocytes stimulated by CMVFFx with or without CsA (1 μg/ml) to restore functional activities of lymphocytes from primary cultures treated or not treated with CsA. IL‐2 significantly enhanced lymphoproliferation, cell‐mediated cytotoxicity to CMV‐infected fibroblasts (CMVF), natural killer cell activity, and the activity of cells capable of suppressing the response of fresh autologous cells to CMVFFx, of cells derived from control and CsA‐treated primary cultures. IL‐2 was found in day‐2 supernatants of control cultures but not CsA‐treated cultures. Day‐2 control supernatants were capable of significantly enhancing proliferation and suppressor cell activity but were less efficient at restoring cytotoxic cell function. Day‐2 supernatants from CsA‐treated cultures were not able to enhance lymphoproliferation or cytotoxic cell function but did induce significant levels of suppressor cell activiiy. The results indicate the presence of different functional mediators in the culture supernatants The ability of IL‐2 to restore lymphocyte effector functions against a clinically important virus may have important therapeutic implications in the treatment of this viral infection in immunodeficiency diseases and in the restoration of immune competence after transplantation.