bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of Blastogenic Factor by Alloantigen‐activated Human Lymphocytes
Authors: PFEFFER P. F. , HIRSCHBERG H.
Published: June 2006

Human suppressor cells generated in mixed lymphocyte cultures (MLC) appear to inhibit both cellular proliferation and the mitogenic effects of blastogenic factor (BF). The results presented here indicate that the inhibition of BF by alloantigen‐activated lymphocytes is not due to decreased lymphocyte proliferation as such, since BF normally produced by non‐proliferating cultures was also suppressed. Moreover, the inhibition was not caused by direct suppression of the BF‐producing lymphocytes, since incubation of BF with alloantigen‐activated lymphocytes neutralized the mitogenic effects of BF. We therefore conclude that activated lymphocytes generated in human MLC directly inhibit or utilize BF. Our experiments also indicate that the activated lymphocytes do not have to proliferate to inhibit BF activity.
Change in the Humoral Response of Athymic Nude Mice with Ageing
Authors: PIGUET P.‐F.
Published: June 2006

The evolution of the serum Ig levels of Balb/c‐nu/nu mice was investigated between 1 month and 12 months of age. An increase as a function of age was observed in all classes and subclasses, which was, expressed in percentage of a nu/+ serum, from 130% to 230% for IgM, from 3% to 24% for IgG1, from 12% to 164% for IgG2a, from 28% to 62% for IgG2b, and from 10% to 50% for IgA. This increase correlates with an increase of plasma cells of each class in the bone marrow, whereas the number of plasma cells in the spleen, the lymph node, and the intestinal mucosa did not change markedly with age. The humoral response after an injection of heterologous erythrocytes was compared in young and aged nu/nu mice; aged mice had a higher haemagglutination titre mainly due to direct (IgM) antibodies. The response of the spleen, as judged by plaque‐forming cells (PFC), was similar in young and aged mice, but the bone marrow response, not detectable in young mice, was about as high in aged nude mice as in nu/+ mice. Although the content of Thy 1 cells in the spleen and lymph nodes was markedly higher in aged than in young nude mice, no T‐cell function could be detected at any age, either in the response to phytohaemagglutinin or concanavalin A or in a graft‐versus‐host assay. Increase in the Ig production with age is interpreted as the result of progressive priming and hyperimmunization by environmental antigens, leading to a T‐independent immune response (even against antigens considered to be T‐dependent) predominantly located in the bone marrow.
Proliferative and Cytolytic Reactivities of Human Lymphocytes Fractionated on Wheat Germ Agglutinin‐Sepharose Columns before or after Alloactivation in Mixed Lymphocyte Culture
Authors: LEHTINEN T. , PERLMANN P. , HELLSTRÖM U. , HAMMARSTRÖM S.
Published: June 2006

Human T‐lymphocyte preparations from peripheral blood with either high‐ or low‐avidity receptors for wheat germ agglutinin (WGA) were obtained by fractionation on WGA‐Sepharose columns. Both fractions contained progenitors of alloreactive T cells, proliferating in mixed lymphocyte culture (MLC) and acting as effector cells in cell‐mediated lympholysis (CML). Proliferation and CML activity of the two fractions were equal and similar to those of the unfractionated cells. However, when the lymphocytes were fractionated after 5 days’ MLC, most of the proliferating and cytolytic cells were found in the lymphocyte fraction enriched in cells with high‐avidity receptors for WGA. The reactivity of the fractions was correlated to their content of blast‐transformed cells. The binding of MLC‐activated lymphocytes to WGA was specific, since it was inhibited by the competitive hapten d‐GlcNAc and no cells were retained on control columns charged with human serum albumin–Sepharose. B cells and monocytic cells were enriched in the fraction with low‐avidity receptors for WGA. As indicated by experiments in which cells were mixed in different proportions, the low MLC–CML activity of the lymphocytes in the fraction with low‐avidity WGA receptors was not caused by suppressor cells present in that fraction.
Production of Human IgM Anti‐D in Tissue Culture by EB‐Virus‐transformed Lymphocytes
Authors: BOYLSTON A. W. , GARDNER B. , ANDERSON R. L. , HUGHES‐JONES N. C.
Published: June 2006

Lymphocytes obtained from two donors who were producing anti‐D were transformed with Epstein‐Barr virus and grown in tissue culture. The lymphoblasts from both donors produced anti‐D antibody, which agglutinated Rh‐positive cells in saline. Both antibodies belonged to the IgM class; the concentration of anti‐D in the culture fluid from one of the donors was approximately 250–500 ng/ml and bound firmly to Rh‐positive cells with a functional affinity constant of approximately 1 × 109 M−1.
Production and Main Characteristics of a Fetal Calf Serum‐specific Cell Line that Induces T and B Cell Differentiation
Authors: RUBIN B. , COOLEY M. A. , LE BORGNE DE KAOUEL C. , TAYLOR R. B. , GOLSTEIN P.
Published: June 2006

Spleen cells from B6 mice injected with fetal calf serum (FCS) could be kept proliferating as a continuous cell line in vitro provided they were cultured in the presence of irradiated syngeneic spleen cells and FCS. Cells in this cell line showed a strong proliferative response when stimulated with concanavalin A (Con A), and they were able to mediate the following functions: (1) they helped the generation of alloantigen‐specific cytotoxic T lymphocytes (CTL) from thymocyte‐spleen cell mixed lymphocyte cultures (MLC), (2) they induced the generation of CTL from normal syngeneic spleen cells in the absence of allogeneic stimulator cells, and (3) they induced normal spleen cells to differentiate into anti‐sheep erythrocyte (SRBC) plaque‐forming cells (PFC), in the absence of SRBC in the cultures. The use of this cell line (called line 12) may thus provide an interesting approach for the study of cellular and molecular requirements for cell‐cell interactions and for the differentiation of T and B effector functions.
Different Macrophage Requirement in the Specific and Polyclonal Responses Induced by TNP‐LPS and LPS
Authors: MARTINEZ‐ALONSO C. , BERNABE R. R. , DIAZ‐ESPADA F.
Published: June 2006

The in vitro anti‐trinitrophenyl (TNP) response induced by TNP‐lipopolysaccharide (TNP‐LPS) in mouse spleen cells was eliminated by passage through Sephadex G10 columns. Conditioned medium obtained from peritoneal macrophages restored the response, indicating the supportive role of such cells in the response. Conversely, polyclonal B‐cell activation mediated by LPS was not affected by Sephadex G10 treatment, as judged by incorporation of 3H‐thymidine, generation of specific anti‐TNP plaque‐forming cells, and induction of polyclonal IgM‐secreting cells. The failure of the LPS moiety of TNP‐LPS to induce B‐cell activation without macrophage help suggests a restriction in the number of available anti‐LPS receptor molecules when the Ig anti‐TNP receptor interacts with the haptenic (TNP) groups. This restriction can be due to the attachment of the TNP molecules to residues located in the vicinity of the lipid A mitogenic region of the carrier.
Effect of CBA/N xid on Spontaneous Production of Antibodies to DNA in MRL/1 and NZB Backcross Mice
Authors: COWDERY J. S. , TAUROG J. D. , STEINBERG A. D.
Published: June 2006

We have used the CBA/N X‐linked B‐cell maturation defect (xid) as a probe to analyse the B‐cell subset involved in spontaneous anti‐DNA production in MRL/1 and NZB mice. The presence of xid in NZB offspring significantly suppressed anti‐DNA production, whereas it did not suppress spontaneously produced antibody to DNA in MRL/1 offspring. These studies suggest that largely non‐overlapping B‐cell subsets are responsible for the spontaneous production of anti‐DNA in MRL/1 and NZB mice.
Senile Cardiac Amyloid is Related to Prealbumin
Authors: SLETTEN K. , WESTERMARK P. , NATVIG J. B.
Published: June 2006

The major component, protein ASC, isolated from the amyloid fibrils in senile cardiac amyloidosis, was characterized by structural studies of some peptic peptides. One of the peptides has an amino acid sequence identical to residues 70–90 in human prealbumin, and three other smaller peptides have a primary structure identical to that in positions 96–107, 109–115 and 121–127. The amino acid composition of the protein resembles that of human prealbumin but shows some characteristic differences. The protein has a blocked N‐terminus and gives no visible precipitin reaction with antiserum to human prealbumin.
Synthesis of Secretory Component by Rat Hepatocytes in Culture
Authors: ZEVENBERGEN J. L. , MAY C. , WANSON J. C. , VAERMAN J. P.
Published: June 2006

Normal rat hepatocytes were isolated and cultivated in vitro. Synthesis of secretory component was demonstrated by its accumulation in the culture medium, as measured by radioimmunoassay; by incorporation of 14C‐leucine in the protein specifically precipitated with anti‐secretory component antiserum; and by a positive precipitin reaction of concentrated culture medium with the same antiserum. The results explain the high levels of secretory component found in rat bile and render plausible a mechanism of hepatic IgA transfer involving secretory component as the hepatocyte membrane receptor for polymeric IgA.
Epstein‐Barr Virus‐induced Lymphoblastoid Cell Lines from Patients with Primary Immunodeficiency Diseases
Authors: TSUCHIYA S. , KONNO T. , TADA K. , ONO Y.
Published: June 2006

Peripheral lymphocytes from eight patients with congenital immunodeficiency diseases were infected with Epstein‐Barr virus (EBV) in an attempt to establish B lymphoblastoid cell lines (LCL). These patients included three boys with congenital agammaglobulinaemia, two girls with hypogammaglobulinaemia, one boy with common variable immunodeficiency, one boy with severe combined immunodeficiency with adenosine deaminase deficiency, and one boy with DiGeorge syndrome. Five of the patients bore no surface immunoglobulins (slg) on their peripheral lymphocytes. LCL were established from seven of the eight patients. All the LCL established formed rosettes with EAC3 and had the ability to produce cytoplasmic immunoglobulins (clg) of various classes. Culture supernatants concentrated up to 100‐fold developed precipitin bands by Ouchterlony's method with antisera to human Ig in all the established LCL. These results suggested that both slg‐, clg‐ and C3+ cells and slg+, clg‐ and C3+ cells might be the target cells for EBV and that slg‐, clg‐ and C3+ cells might be the precursor cells of B lymphocytes.
Proliferation of C3HXCBA Hybrid Lymphocytes in the Spleens of Irradiated CBA Mice: Linkage to the Mls‐Gene
Authors: LILLIEHÖÖK B.
Published: June 2006

Lymphocytes from C3HXCBA F1 mice, lacking delectable Mls‐antigens, can specifically inactivate T‐cell clones from the H‐2‐idcmical strain CBA, which are reactive against the C3H‐Determined Mls‐antigen. Such F1 lymphocytes also proliferate in the spleens of irradiated CBA mice. The age dependence of the mixed lymphocyte culture stimulatory capacity and the capacity to proliferate in irradiated CBA recipients of lymphocytes from C3HXCBA F1 mice were compared. It was observed that appearance of Mls‐bearing cells is preceded by the appearance of F1 cells with strong proliferate capacity in irradiated CBA recipients. The in vivo proliferative response is also dependent on the age of the recipient. The varying response is not reflected in the quantity ofanli‐C3H Mls‐reactive cells in CBA mice. A strong linkage between the genes controlling the above proliferative and stimulatory capacities of C3HXCBA lymphocytes was found.
The Influence of Serum on Random Migration and Chemotaxis of Polymorphonuclear Leucocytes: Methodological Evaluation using Sera from Infection‐prone Patients and Normals
Authors: HÅKANSSON L. , VENGE P.
Published: June 2006

The effects on normal polymorphonuclear leucocyte (PMN) of chemotactic and chemokinetic factors in sera from normal subjects and Infection‐prone patients wan examined by means of the leading‐from technique, using a modified Boyden chamber. The analysts schema used made it possible to differentiate between chemotactic and chemokinetic factors and demonstrated that different factors account for the major chemotactic and chemokinetic activities of serum. The major chemotactic activity of unactivated serum was heat‐labile, and the chemotactic activity of factor XII‐deficient sera was normal, suggesting the major chemotactic activity to be distinct from C5a and factor XII‐dependent pathways. The existence of both heat‐stable and heat‐labile chemokinetic factors was shown. The possibility that the reduced chemokinetic effect of several patient sera was caused in abnormal levels of the serum proteins γ1‐antitrypsin, γ2macroglobulin and albumin was excluded.
Spontaneous Plaque‐forming Human Lymphocytes Detected with the Protein‐A Plaque Assay
Authors: FREIJD A. , KUNORI T.
Published: June 2006

A sizable proportion of fresh human blood and adenoid lymphocytes were found to produce antibodies when tested in the protein‐A plaque assay of Gronowicz et al. Plaque formation was not caused by release of passively adsorbed immunoglobulin but depended on active‐ immunoglobulin secretion We have Investigated optimal conditions for plaque formation In this test system. Good reproducibility is obtained only if Cells to be tested are kept in the cold before assay, since the antibody‐forming capacity of these cells is rapidly exhausted at temperatures above 2°C. In blood lymphocytes the number of IgA‐producing cells exceeds those of IgG and IgM, whereat in the adenoid IgG plaque‐forming cell dominate. Studies on the on of this method for the monitoring of immune responses are in progress.
Expression of HLA‐ABC and ‐DR Locus Antigens on Human Kidney, Endothelial, Tubular and Glomerular Cells
Authors: HÄYRY P , VON WILLEBRAND E. , ANDERSSON L. C.
Published: June 2006

The distribution of the major histocompatibility complex antigens on the various cellular structures of the human kidney was analysed by using a modified Staphalococcus aureus Cowan I method Conventional alloantisera and heterologous antisera raised against Isolated molecules were used for the HLA‐ABC antisera were expressed on all types of kidney passenger leucocytes, on vascular endothelial cells, and on kidney tubular cells, but not substantially on the glomerular podocytes, The DR antigens were strongly expressed on (a fraction of) the passenger lymphocytes and on the kidney vascular endothelial cells, weakly on the passenger monocytes, but not measurably on the urine‐producing apparatus of the kidney—that is, on the glomerular and tubular cells.
Correlation between Suppression of B Cell Anti‐Hapten Response and Generation of Cytotoxic T Cells Induced by Hapten‐labelled Syngeneic Lymphocytes
Authors: RAMOS T. , MÖLLER E.
Published: June 2006

The correlation between suppression of B cell antibody response and generation of cytotoxic T cells induced by hapten (FITC) labelled syngeneic lymphocytes was studied. Primary in vitro sensitization with lymphocytes labelled with the hapten concentrations (FITC 0.5 and FITC 0.05) previously shown to induce specific B cell unresponsiveness generated cytotoxic cells that specifically lysed haptenated syngeneic blast cells. In vivo priming with either SC‐FITC 0.5 or SC‐FITC 0.05 could be demonstrated after in vitro restimulation with similarly haptenated cells. Thus, cells from primed mice gave a cytotoxic response already after 3 days in culture, whereas primary responses were only observed on day 5. Furthermore, immunization was evident as an increased specific lysis at low effector/target ratios. However, no direct cytotoxicity could be detected in spleen cells from immune animals. Comparison between kinetics of in vivo development of specific antibody suppression and induction of cytotoxic cells detectable after in vitro boosting revealed no correlation between the two phenomena.
Kinetics of Phagocytosis and Intracellular Killing of Staphytococcus aureus and Escherichia coli by Human Monocytes
Authors: LEIJH P. C. J. , VAN DEN BARSELAAR M. TH. , VAN FURTH R.
Published: June 2006

Tie kinetic patterns of the phagocytosis and intracellular killing of Staphytococcus and Echerichia coli by monocytes were investigated separately to acquire more insight into the total process, i.e. from the ingestion to the death of the micro‐organisms. Phagocytosis proved to be dependent on: (1) both the bacteria‐to‐monocyte ratio and the monocyte concentration; a concentration of at least 5 × 105 monocytes/ml proved necessary for the measurement of ingestion, whereas the rate of ingestion was found to be proportional to the number of extracellular bacteria until a maximum rate is reached, (2) the serum concentration in the incubation medium, which influenced both the rate of phagocytosis and the maximum number of bacteria taken up by one monocyte, and (3) the temperature, the highest rate of phagocytosis being reached at 37–41°C The intracellular killing proved to be dependent on: (1) the number of bacteria ingested; the rate of killing was proportional to the number of ingested bacteria until a maximum rate was reached; (2) the temperature, since a maximum rate of killing is only reached at 37–41°C: at tower and higher temperatures the rate of killing is lower, in the latter case due to inactivation of extracellular stimuli. These separate data on the ingestion and killing processes made it possible to compute the theoretical numbers of extracellular, viable intracellular, and total intracellular bacteria for a model system consisting of 5×106 monocytes, 5×106 bacteria, and 10% serum. These calculated values are in agreement with the experimental data.
Influence of Tonsillar Disease on the Expression of J Chain by Immunoglobulin‐producing Cells in Human Palatine and Nasopharyngeal Tonsils
Authors: KORSRUD F. R. , BRANDTZAEG P.
Published: June 2006

A significant reduction of the percentage of J‐chain‐positive intra‐ and extra‐follicular IgA immunocytes was found in inflamed palatine tonsils. There was a tendency to similar alterations in hypertrophied adenoids. Tonsillar disease apparently enhances local maturation of the B‐cell system, perhaps on the basis of intensified proliferation of memory clones. Alternatively, there may be a disease‐associated defect in the mechanism(s) that normally induces switchover to the IgA isotype early in clonal development. It is speculated that, by decreasing the J‐chain expression during local B‐cell differentiation, tonsillar disease may jeopardize the potential of the tonsils as a putative precursor source for the secretory immune system of the upper aero‐digestive tract.
Immunoglobulin Expression in Human Lymphoblastoid Cell Lines with Early B Cell Features
Authors: GUGLIELMI P. , PREUD'HOMME J. L.
Published: June 2006

Immunoglobulin expression was studied by direct immunofluorescence and by biosynthesis experiments in two human cell lines Raji and T 5.1. The basic phenotype of these cells was close to that of pre‐B calls: large cell with intracytoplasmic IgM with a predominance of μ chains over light chains and no detectable surface immunoglobulins. The apparent molecular weight of heavy and light chains was abnormally large. Immunoglobulins were secreted at a low rate as pentameric IgM in the T 5.1 line and as subunits and free light chains in the Raji line. Spontaneous variations of this phenotype were observed: the cultured edit acquired surface μ and λ chains, then additionally δ chains while they progressively lost delectable cytoplasmic μ chains, thus leading to a mature B cell phenotype, Subsequently, the cells had no detectable surface and cytoplasmic immunoglobulins and then they displayed a pre‐B cell phenotype again. Attempts to induce further maturation using various potential inducers were unsuccessful
Intestinal, Salivary, and Tonsillar IgA and J‐Chain Production in a Patient with Severe Deficiency of Serum IgA
Authors: BRANDTZAEG P. , GUY‐GRAND D. , GRISCELLI C.
Published: June 2006

An 18‐year‐old man with tendency to respiratory infections had a serum IgA level of only 2% of normal whereas his salivary IgA amounted to 50% of the lower normal concentration range. Moreover, both the rectal and jejunal IgA‐producing cell populations were of normal size. Nevertheless, a relative increase of salivary IgM and a distinctly raised number of IgM‐producing cells in jejunal mucosa indicated an imbalance in his secretory immune system. This possibility was supported by the presence of an excess of J chain in most of his intestinal IgA immunocytes, probably reflecting a reduced synthetic rate of IgA. The number of tonsillar IgA‐producing cells was only slightly below the normal range: most of them lacked J chain, as normal, and could thus be a source of his serum IgA, which was mainly monomeric. A marked deficiency of IgA‐producing cells in his hone marrow supported the notion that this tissue site normally is the major source of monomeric IgA. This study suggests that a generally defective IgA system may he topically activated owing to the persistent antigenic and mitogenic load on mucosa‐associated lymphoid tissues. Our findings are not consistent with a general regulative compartmentalization of monomer‐ and dimer‐producing IgA immunocyte populations.
Ia Antigens on Antigen‐presenting Cells do not Carry the Same Ia Specificities as Detected on Suppressor and Helper T Cells
Authors: ASANO Y. , OKUMURA K. , TADA T.
Published: June 2006

The passage of spleen cells through a tightly packed nylon wool column partially separated two populations of cells required for the antigen‐induced secondary proliferative response of T cells. The proliferating T cells were in the nylon wool column‐passed cell population, but the addition of a small number of nylon‐wool‐adherent cells was required for their maximal proliferative response. Such adherent cells were non‐T, non‐B, Ia antigen‐positive cells serving as antigen‐presenting cells. Ia determinants on this cell type are encoded in two separate I subregions, I‐A and I‐E/C, and they are expressed simultaneously on the same cells. Antisera specific for I‐J subregion gene products having as a known activity the killing of antigen‐specific suppressor and helper T cells were unable to kill antigen‐presenting adherent cells. The results indicated that at least I‐J subregion gene products expressed on suppressor and helper T cells are not present on antigen‐presenting accessory cells. The Ia specificities detected on accessory cells follow the cross‐reactive pattern found in Ia molecules of B cells, and thus antigen‐presenting cells carry Ia antigen identical to those of B cells, and not those of T cells.
Treatment of Human Complement Components C4 and C3 with Amines or Chaotropic Ions
Authors: VON ZABERN I. , NOLTE R. , VOGT W.
Published: June 2006

Treatment of human components C4 and C3 with amines like hydrazine, ammonium hydroxide, and neutral ammonium salts or with chaotropic salts like KSCN und NaBr leads to complete loss of haemolytic activity. The pretreated components are however, still active in formation of soluble C3 convertases. This activity pattern is reminiscent of the activities of C4 and C3 that have been activated by cleavage in the fluid phase. Indeed, the antigenic properties of pretreated C4 and C3 are similar to soluble C4b and C3b. The polypeptide chain structure of pretreated C4 and C3, is, however, identical to that of the untreated components when investigated by SDS gel electrophoresis. Pretreatment even reduces greatly the susceptibility of C4 to cleavage by C1 and of C3 to cleavage by classical and alternative pathway C3 convertases. Pretreated components have lost the ability to combine with EAC1 and EAC142, respectively; this fact explains their failure to exhibit haemolytic activity. In serum, pretreated C4 and C3 are cleaved in a manner similar to C4b and C3b. Amines and chaotropic ions cause the same functional and structural alterations, which are best explained by assumption or a conformational change. A similar transformation can also occur in C4 and C3 during preparation or storage
Fc Receptors on Human T Lymphocytes: Loss of Fcμ and Expression of Fcγ Receptors by T Cells Stimulated in Mixed Lymphocyte Reaction
Authors: MORETTA A. , MINGARI M. C. , COLOMBATTI M. , MORETTA L.
Published: June 2006

The consequences of allogeneic stimulation on the expression of Feμ or Feγ receptors were analysed in T‐cell populations responding in mixed lymphocyte reaction (MLR). Unfractionated T‐cell populations were cultured with a pool of irradiated allogeneic T‐depleted cells. The responder E ‐rosetting cells progressively lost Feμ and acquired Fcγ receptors. The change of the original Fc receptor phenotype is not the consequence of a preferential proliferative response of TG versus TM cells but is likely due to a de novo expression of Feγ receptors by T cells lacking detectable Fc receptors (T‐null) and also to the loss of Fcμ and expression of Feγ receptors by TM cells. These data suggest that, after MLR, responder T cells can modify their Fc receptor phenotype.
The Mouse Primed Lymphocyte Typing (mPLT) Test
Authors: PECK A. B. , KLEIN J. , WIGZELL H.
Published: June 2006

The mouse primed lymphocyte typing (mPLT) assay, based on the sequential reactivation of specific immunocompetent, alloantigen‐reactive T blast cells in secondary mixed leucocyte culture (MLC), has been utilized to define the major histocompatibility complex (MHC) Class I‐associated lymphocyte‐stimulating (LS) determinants. The test panel of secondary stimulating cells has been expanded to include 25 B10.W lines (mouse strains carrying MHC regions derived from wild mice), thus permitting examination of more than thirty ‘independently derived’ MHC haplotypes. Data obtained using mPLT cells generated in primary MLC between H‐2 K‐ and/or DL‐disparate strain combinations indicate that each allelic Class I product expresses a unique set of LS determinants recognized by alloreactive T lymphocytes. In addition, we have observed that Class I products, which are apparently serologically identical but which are encoded for by genes of independently derived haplotypes, can express mutually distinct LS determinants. These data suggest that Tlymphocytes possess the capacity to distinguish between Class I antigens that are serologically similar (or even identical) and support the concept that LS determinants recognized by T cells are distinct from the majority of specificities recognized by B cells. This concept has marked importance in interpretation of data in the human system, for which there is now an attempt to correlate serology and PLT.
Regulation of Effector Functions of Human K‐Cells and Monocytes by Antigen Density of the Target Cells
Authors: ÖHLANDLR C. , LARSSON Å. , PERLMANN P.
Published: June 2006

Antibody‐dependent cytolysis and phagocytosis mediated by human K‐cells or monocytes against chicken erythrocytes (ChRBC) were studied. The antigen density of the target cells was varied by coating the cells with different amounts of 3H‐labelled dinitrophenyl (DNP) hapten. The degree of antigenicity thus acquired by the target cells was assessed on the basis of their uptake of the isotope. Anti‐DNP scrum was used to induce lysis or phagocytosis. Below 500 antigenic determinants per ChRBC the target cells were not affected. However, at that density, lysis and/or phagocytosis was seen when the antibody concentration was high (2 × 10−9m). With less antibody present (2 × 10−11m) only monocyte‐mediated phagocytosis was induced. The estimated lowest number of target‐cell‐bound antibodies required for K‐gel I‐mediated lysis was ≃ 50. The corresponding number for monocyte‐mediated phagocytosis was ∼20 IgG per ChRBC. The result suggests that interaction of several Fc receptors on the effector cells with IgG molecules bound to adjacent sites on the target cell membrane is an important factor in the regulation of these antibody‐dependent cell‐mediated effector functions.
Inhibition of de Novo IgM Antibody Synthesis by Thalidomide as a Relevant Mechanism of Action in Leprosy
Authors: SHANNON E. J. , MIRANDA R. O. , MORALES M. J. , HASTINGS R. C.
Published: June 2006

Thalidomide is well documented to be an effective treatment for erythema nodosum leprosum (ENL) occurring in lepromatous leprosy. To be beneficial, thalidomide must interfere with one or more of the several essential steps in the pathogenesis of this syndrome, which is presumed to be a clinical manifestation of an Arthus‐type hypersensitivity. Since complexes of antigen and antibody would initiate these events, thalidomide could exert its most direct influence on reactants in this essential step. To determine whether thalidomide affected de novo antibody synthesis, the effect of the drug on the antibody response to sheep erythrocytes in mice was determined. Thalidomide significantly inhibited IgM antibody formation when fed to mice for 5 or 7 days before immunization with sheep erythrocytes. There was also a selective decrease in serum IgM concentrations among leprosy patients being treated with thalidomide for ENL. A clinically relevant site of action of thalidomide in ENL appears to be on the synthesis of IgM antibody. The target site of the drug among the macrophage, antibody‐forming, and helper or suppressor lymphocytes remains to be elucidated.
Conditions for Cytomegalovirus Stimulation of Lymphocytes
Authors: WAHREN B. , ROBÈRT K.‐H. , NORDLUND S.
Published: June 2006

Lymphocytes from healthy donors or from patients with chronic lymphocytic leukaemia were subjected to live or inactivated cytomegalovirus (CMV) or the mitogen phytohaemagglutinin. No early or late CMV antigens could be demonstrated in the lymphocytes, indicating that neither abortive nor replicative CMV infection takes place. Only cells from CMV antibody‐positive leukaemic and non‐leukaemic donors were stimulated by CMV to DNA synthesis, with a maximum on day 5. Cells from all individuals responded to phytohaemagglutinin stimulation, the peak of activity occurring on day 3. The stimulation with CMV occurred in T cells and was independent of early CMV antigen production, viral DNA synthesis, or viral replication. CMV is thus not an in vitro lymphocyte mitogen like Epstein‐Barr virus but is a very potent antigen for memory T cells.
Neocortical Lateralization of NK Activity in Mice
Authors: BARDOS P. , DEGENNE D. , LEBRANCHU Y. , BIZIÈRE K. , RENOUX G.
Published: June 2006

The natural killer (NK) reactivity of mouse spleen cells is controlled by the left brain neocortex and not by the right symmetrical brain area. The finding strongly suggests direct relationships between the central nervous system and the immune system, both involved in biological adaptation for the maintenance of homeostasis and body integrity in relation to the external environment.
Distribution of Surface, Cytoplasmic and Secreted IgG Subclasses in Human Lymphoblastoid Cell Lines and Normal Peripheral Blood Lymphocytes
Authors: SIMMONS J. G. , FULLER C. R. , BUCHANAN P. D. , YOUNT W. J.
Published: June 2006

Twenty‐two IgG‐positive human lymphoblastoid cell lines and normal peripheral blood lymphocytes were studied for surface and cytoplasmic IgG, IgG subclasses, IgD and IgM, using monospecific fluorescein‐and rhodamine‐conjugated F(ab′)2 antibody fragments, and for secretion by double antibody radioimmunoassay. Several parallel observations and several differences in IgG subclass expression were noted between cell lines and normal lymphocytes. Surface IgG2 was frequently expressed in normal IgG‐positive lymphocytes but was seldom expressed in cell lines. Cell lines resembled normal IgG‐positive lymphocytes in the frequent expression of cytoplasmic IgG3 and lgG4, often without secretion. Cell lines and normal lymphocytes both showed more frequent distribution of IgG and IgG sub‐classes in cytoplasm than in surface immunoglobulin, and often a discrepancy of surface versus cytoplasmic IgG subclass. A good correlation was noted between surface, cytoplasmic and secreted IgG1. Despite a predominance of IgG2 and IgG4 surface IgG subclasses, and IgG3 and IgG1 in cytoplasm, secreted immunoglobulins from normal lymphocytes in short‐term culture showed a similar distribution of IgG subclasses to that seen in normal sera. Multiple expression of IgG subclasses was much more frequent in IgG‐positive cell lines than in normal peripheral blood lymphocytes, both in surface and cytoplasmic IgG.
Promotive and Inhibitory Effects of Macrophages on the Mitogen‐induced Blastogenesis of Lymphocytes: A Flow‐cytometric Study
Authors: STEEN H. B. , MUNTHE‐KAAS A. C.
Published: June 2006

The effect of various concentrations of rat‐liver macrophages on the Con A‐induced blastogenesis of syngeneic spleen lymphocytes has been studied by means of flow‐cytometric methods and by measurement of the uptake of 3H‐thymidine. By measuring the distributions of cellular and nuclear volume and cellular DNA to characterize the progression of responding lymphocytes through the cell cycle, we have distinguished a promotive and an inhibitory effect of macrophages. The promotive effect is on the number of cells initiating blastogenesis and on their rate of progression through the cell cycle. The degree of promotion increased strongly with the concentration of macrophages even for concentrations that suppressed the incorporation of 3H‐thymidine almost completely from about 30 h of culture. The inhibition observed for macrophage concentrations ± 10% was a late effect causing stagnation of cell cycle and reduced viability only from about 24 h of culture.
The Generation of Specific Cytotoxic Responses by Human Lymphocytes Requires Antigen and Activities Provided by a Lymphokine Supernatant‐
Authors: WARREN H. S.
Published: June 2006

The requirements for the generation of specific cytotoxic responses by human lymphocytes have been investigated using as a model system the response to the ultraviolet light‐killed mouse tumour P815. These requirements are antigen (P815uv) and activities provided by a human lymphokine (LK) supernatant. Cells that give rise to non‐specific cytotoxicity can be separated on discontinuous Percoll density gradients after a 3‐day incubation of peripheral blood leucocytes with the lymphokine supernatant. Cells from which specific cytotoxic responses can subsequently be generated in the presence of P815uv and LK sediment at a higher density.
Albumin‐Immunoglobulin Complexes in Human Serum: Classification and Immunochemical Analysis
Authors: EILAT D. , FISCHEL R. , ZLOTNICK A.
Published: June 2006

A complex between serum albumin and immunoglobulins was observed on immunoelectrophoresis in six patients. Two patients with multiple myeloma had monoclonal IgA–albumin complexes; one of these complexes was formed by covalent bonds and the other by non‐covalent bonds. Four patients displayed non‐covalent IgG‐albumin complexes: of these, one had multiple myeloma, two had been treated with nitrofurantoin for prolonged periods, and one had diabetes mellitus. The IgG‐albumin complex of the last patient was subjected to a detailed immunochemical analysis. The albumin‐specific antibodies were isolated by affinity chromatography and analysed, using a sensitive tritium labelling technique. The antibodies were polyclonal, complexed with serum albumin through their Fab portion, and showed a high specificity for the human albumin as compared with bovine and rabbit albumins. The serum albumin of two patients displayed an abnormal behaviour in reduced SDS‐polyacrylamide gel electrophoresis (PAGE). The abnormal albumins had an apparent molecular weight of 52,000 and could read with rabbit anti‐human serum albumin like the normal protein. No abnormal albumin could be detected in 20 other patients' sera, including nitrofurantoin‐treated patients and normal controls. These findings suggest a possible role for an altered self‐component in the triggering of a specific autoimmune reaction.
Human Blood Monocytes: Characterization of Negatively Selected Human Monocytes and Their Suspension Cell Culture Derivatives
Authors: STEVENSON H. C. , KATZ P. , WRIGHT D. G. , CONTRERAS T. J. , JEMIONEK I. F. , HARTWIG V. M. , FLOR W. J. , FAUCI A. S.
Published: June 2006

Normal human monocytes were negatively selected from leucapheresis cell suspensions by countercurrent centrifugation–elutriation in high yield with a mean purity of 93.5%. The combination of the novel methods of negative cell selection and suspension cell culture has provided the opportunity to study serially over several days the morphologic and functional changes of monocytes from a single donor as they matured in culture to typical macrophages. Human monocytes nearly double in size during the first week of culture, experiencing near daily increases in cell volume. This was associated with changes in the ultrastructure of these cells, including the development of numerous small knob‐like projections on the cell membrane and the proliferation of microtubules and filamentous Structures within the cell cytoplasm during the first 6 days of culture. Peroxidase activity declined during the first 4 days of culture, whereas 5′‐nucleotidase activity was acquired during the first 48 h of culture. Lysozyme activity in the cultures increased from day 2 to day 6 of culture. The phagocytic capacity of monocytes for IgG‐coated erythrocytes increased dramatically during the first week of culture, but the cytotoxic capability of monocytes against similar targets in an antibody‐dependent cytotoxicity assay declined to nearly hall of base‐line levels by day 2 of culture and remained at this diminished level during subsequent days in culture.
Adoptive Transfer of Delayed‐Type Hypersensitivity to Sendai Virus
Authors: ERTL H. C. J.
Published: June 2006

The intrathymic versus extrathymic influences on H‐2‐restriction specificity of K.D‐ and I‐region‐restricted Sendai virus‐immune T lymphocytes, which mediate a delayed‐type hypersensitivity response in vivo, were investigated. K,D‐ and I‐region‐restricted Td lymphocytes of P1→F1 and F1→P1 fetal liver chimeras showed preference but no absolute restriction for the P1 haplotype. T effector lymphocytes of thymic chimeras expressed a slightly higher degree of restriction for the haplotype of the thymus graft than F1→P1 chimeras, which might reflect the activity of T lymphocytes of thymic origin. The lack of absolute restriction of both K,D‐ and I‐region‐restricted Td lymphocytes to the haplotype the thymus suggests an additional pathway of T‐cell maturation.
Interferon‐Induced NK Augmentation in Humans
Authors: ULLBERG M. , MERRILL J. , JONDAL M.
Published: June 2006

By combining a single‐cell cytotoxicity assay in agarose with estimations of the maximal natural killer (NK) cell potential (Vmax) by 51Cr release, the mechanisms behind interferon augmentation of human NK cells were analysed. The number of target‐binding cells (TBCs) the fraction of active TBCs, and NK cell recycling were studied after short‐term interferon treatment. The results demonstrate a dual effect of interferon on human NK cells: effector cell recruitment and increased effector cell recycling. Both of these variables were increased when NK cells were tested against the standard target K‐562 and against Daudi and BJAB cells, derived from B‐type lymphomas. However, when T cell lines derived from acute lymphocytic leukaemia (Molt‐4 and 1301) were used as targets, a larger fraction of active NK cells were found among untreated TBCs, whereas inteferon treatment only resulted in increased effector cell recycling and not in effector cell recruitment. No increase in TBCs after interferon treatment could be detected with any cell line tested. The difference seen between T and non‐T cell lines with regard 10 interferon‐induced effector cell recruitment is discussed in relation to known characteristics of the human NK system.
Electrophoretic Polymorphism of Human C4 is Due to Charge Differences in the α‐Chain, Presumably in the C4d Fragment
Authors: MEVÅG B. , OLAISEN B. , TEISBERG P.
Published: June 2006

Various common C4 gene products were isolated from serum by immunoprecipitation. After reduction the C4 α‐, β‐, and γ‐polypeptide chains were studied by two‐dimensional electrophoresis. Isoelectrofocusing was performed in the first dimension and sodium dodecyl sulphate polyacrylamide gradient gel electrophoresis in the second. The charge differences behind the electrophoretic C4 polymorphism were shown to reside in the 95,000‐u(atmic mass units) α‐chain. Charge variation closely mirroring the α‐chain differences were also found in a 49,000‐u fragment of the α‐chain, most probably C4d. The basic β‐chain could not be studied in detail, but no differences were observed with regard to molecular weight or charge of the γ‐chains of the different C4 gene products.
HLA‐DR Antigens Render Interleukin‐2‐Producer T Lymphocytes Sensitive to Interleukin‐1
Authors: PALACIOS R.
Published: June 2006

Monoclonal anti‐HLA‐DR antibodies inhibited the production of interleukin‐2 (IL‐2) when added from the initiation of autologous (AMLR) and allogeneic (MLR) mixed lymphocyte reactions, but not 60 h later. The inhibitory activity of the anti‐DR sera became apparent 8 h after initiation of the cultures and was maintained throughout the culture period. Interleukin‐1 (IL‐1) added to cultures carried out in the absence of the anti‐DR antibodies significantly enhanced the production of IL‐2, whereas addition of IL‐1 to anti‐DR‐treated AMLR and MLR cultures did not restore or increase the synthesis of IL‐2. However, when the anti‐DR antibodies were added to IL‐1‐supplemented AMLR and MLR cultures 60 h or more after initiation of the reactions, the antiserum no longer inhibited the capacity of IL‐1 to promote the synthesis of IL‐2 or the production of IL‐2. Finally, resting T cells were unresponsive to IL‐1 and did not produce IL‐2. It thus seemed that the anti‐DR antibodies inhibited production of IL‐2 in AMLR and MLR by rendering the IL‐2 producer T cells unresponsive to IL‐1. Cyclosporin‐A, a drug that abrogates activation of T cells by blocking their receptors for HLA‐DR antigens, also rendered IL‐2‐producer T cells unresponsive to IL‐1 and abrogated the production of IL‐2 in AMLR and MLR. Since resting T cells do not respond to IL‐1 or produce IL‐2, it is concluded that HLA‐DR antigens of the stimulator cells participate in the production of IL‐2 in AMLR and MLR by enabling the IL‐2 producer T lymphocytes to respond to IL‐1. Interleukin‐1 promotes the production of IL‐2 by IL‐1‐sensitive T cells. Once the IL‐2‐producer T cells become sensitive to IL‐1, there is no further requirement for HLA‐DR antigens.
Acidic Protein in Macrophages
Authors: PERSSON L. I. , HANSSON E. , RÖNNBÄCK L.
Published: June 2006

An antiserum to unstimulated rat peritoneal macrophages was produced in rabbits. The antibodies were directed against an acidic protein with a molecular weight of 35,000 and with an isoelectric point at 4.6. The macrophage acidic protein (MAP) was purified by gel filtration of rat lung soluble proteins, followed by preparative isoelectric focusing. The preparation of MAP was pure as assayed by agar gel electrophoresis and showed one precipitation peak in crossed immunoelectrophoresis against the crude antiserum directed against peritoneal macrophages. The purified MAP was used for immunization of rabbits, and the antiserum obtained was monospecific, assayed by crossed immunoelectrophoresis and Grabar‐Williams immunoelectrophoresis. The titre was 4 times higher in the anti‐MAP antiserum (1:80) than in the crude antimacrophage antiserum (1:20), tested against MAP by counter‐current immunoelectrophoresis. The antigen (MAP) was demonstrated by direct and indirect immunofluorescence microscopy in rat blood monocytes, in spleen and lung monocytic cells, in dusters of cells in the thymus, and in adventitial macrophages around larger blood vessels in liver, kidney, lung and brain. Scattered meningeal macrophages showed fluorescence in the normal brain. In stab‐wounded areas of rat brain MAP was localized to perivascular and perineuronal macrophages with a morphology similar to that of microglial cells. The localization of the fluorescence was the same both for the anti‐serum against MAP mid for the antiserum raised against crude peritoneal macropages.
Characterization and Possible Mechanisms of Mitogen‐induced Cell‐mediated Cytotoxicity
Authors: YUE C.‐L. , TANIMOTO K. , HORIUCHI Y.
Published: June 2006

Mitogen‐induced cell‐mediated cytotoxicity (MICC) of human peripheral blood granulocytes (PMN) and lymphocytes (PBL) was studied by using chicken erythrocytes as targets. The possible mechanisms were elicited bidirectionally by means of the MICC inhibition test and by functional analysis of lectin. In our preliminary data it was noted that PMN‐mediated cytotoxicity was more potent than that of PBL. In addition, erythrophagocytosis or lysosomal enzymes released from PMN offered little contribution to cytotoxic activity of PMN. From the inhibition study it was realized that intact cytoskeletal structures, functionally preserved membrane lectin receptors, active DNA synthesis, and environmental divalent cations were mandatory for the full expression of MICC activity by both effectors. We also identified the cytotoxic activity of PHA‐P as having a unique character that is thermostable and independent of the other biological activities.
Effects of Thymosin and Evidence of Monocyte Suppression of Both T‐ and B‐Cell Functions in Two Cases of ‘Common Variable Immunodeficiency’
Authors: ARALA‐CHAVHS M. P. , KORN J. H. , GALBRAITH G. M. P. , PORTO M. T. , SMITH C. L. , FUDENBFRG H. H.
Published: June 2006

We have studied two patients with common variable immunodeficiency (CVID) impaired cell‐mediated immunity. and high percentages of monocytes in their peripheral blood. Removal of monocytes from cultures of peripheral blood mononuclear cells from both patients increased the in vilro responses to phytohaemagglutinin (PHA) and concanavalin A (Con A) but not to purified protein derivative (PPD), as measured by [3H]thymidine uptake. Similarly, supernatants of monocyte cultures from both patients. unlike supernatants of normal monocytes, suppressed the in vitro responses to PHA and Con A but enhanced the response to PPD by cultured mononuclear cells from the patients and from normal donors. Addition of unfractionaicd mononuclear cells from both patients 10 normal mononuclear cells suppressed both pokeweed mitogen (PWM) stimulation and IgG production: this effect was abrogated by removal of monocytes from the patients’ mono‐nuclear cell populations. The effect of thymosin on both patients’ mononuclear cells was assayed in vitro. Thymosin was ineffective in vitro with cells from the first patient: for the other patient. [3H]thymidine uptake by mononuclear cells stimulated with PPD increased. whereas uptake by Con A‐stimulated cells decreased, as did the percentage of E rosette‐forming cells, providing further evidence of heterogeneity of the CVID syndrome. The effects of thymosin were also dependent on monocytes.