bluuebunny/crossref_metadata_2025 huggingface dataset:
Effector Mechanisms in Allograft Rejection III. Kinetics of Killer Cell Activity Inside the Graft and in the Immune System During Primary and Secondary Allograft Immune Responses
Authors: ROBERTS P. J.
Published: June 2006

The cells recovered from sponge matrix allografts and from various lymphoid organs of recipient mice were tested for their ability to perform relevant target cell lysis in vitro. In the primary response, the peak cytotoxic activity was detected 8 days after transplantation, inside the graft, in regional lymph nodes, and in the spleen. In the secondary response, the peak cytotoxic activity in the regional lymph nodes and in the spleen took place promptly, 4 days after transplantation. The peak activity inside the graft was recorded on day 7. In the primary response, the activity in the blood was low, and significant cytotoxic activity appeared only after the activity inside the graft had declined. In the secondary response, the cytotoxic activity in the blood was high and demonstrated a biphasic profile, with one peak on day 3–4 and another activity peak on day 14. Killer cells were detected inside the graft for a far longer time in she primary (4 weeks) than in the secondary (2 weeks) response. I conclude that cellular kinetics in the secondary anti‐allograft response is qualitatively but possibly not quantitatively different from the primary response.
Differences in the Mode of Phagocytosis with Fc and C3 Receptors in Macrophages
Authors: KAPLAN G.
Published: June 2006

Mouse macrophages, activated in vivo or in vitro, were made to internalize sheep erythrocytes opsonized with IgG or IgM and complement. Scanning electron microscopy and transmission electron microscopy were performed ac various times after the transfer of the cultures from conditions favoring attachment to conditions favoring internalization. The receptors for Fc and C3 were distributed randomly over the macrophage surface with the exception of the extreme periphery, were Fc receptors were more abundant. The E‐IgG were ingested by means of thin membrane extensions rising from the macrophage surface and enclosing the opsonized particles tightly in a cup‐like structure protruding from the macrophage surface. Only afterwards were the covered particles drawn into the cell body proper. The E‐IgMC were seen to sink directly into the macrophage cytoplasm without apparent involvement of membrane extensions. Experiments with cytochalasin B suggested that microfilaments were essential for the phagocytosis by Fc but much less important with the C3 receptor.
IgD and the Fetal Immune Response
Authors: CEDERQVIST L. L. , EWOOL L. C. , LITWIN S. D.
Published: June 2006

IgD was detected in 84% of cord sera at term by an inhibition‐of‐hemagglutination test, in contrast to low incidences reported earlier. Cord blood IgD and maternal serum IgD values were unrelated to each other, consistent with a fetal origin of the IgD. The cord blood IgD failed to correlate, either directly or inversely, with cord blood IgA or IgM, suggesting that its presence does not reflect fetal synthesis of specific antibody but rather may be developmental.
Direct Activation of Guinea Pig Macrophages by a B‐Cell Mitogen
Authors: FERNÁNDEZ C , CLERICI N. , KREISLER J. M.
Published: June 2006

Since polyclonal B‐cell activators (PBA) require macrophages to induce modifications in the lymphocyte in vitro primary response to thymus‐dependent antigens, we have investigated whether PBA act directly on macrophages. [14C]glucosamine uptake by guinea pig peritoneal adherent cells after stimulation with purified protein derivative (PPD), lipopolysaccharide (LPS), and dextran sulfate (DxS) was tested. PPD produced an increased [14C]glucosamine uptake, whereas LPS and DxS did not. According to our experiments, (a) PPD docs not require the presence of lymphocytes to stimulate macrophages; furthermore, when lymphocytes were present in a concentration higher than 5%, a suppressor effect in the glucosamine uptake was found, and (b) there was no significant difference between the findings when peritoneal adherent cells were cultured in normal medium and in supernatant of lymphocyte cultures stimulated with PPD.
Complement Lysis: the Ultrastructure and Orientation of the C5b‐9 Complex on Target Sheep Erythrocyte Membranes
Authors: TRANUM‐JENSEN J. , BHAKDI S. , BHAKDI‐LEHNEN B. , BJERRUM O. J. , SPETH V.
Published: June 2006

The C5b‐9 complex derived from human serum and assembled on target sheep erythrocyte membranes is a thin‐walled cylinder rimmed by an annulus at one end. The total height of the cylinder is 150 Å, towards which the annulus contributes 30 Å. The cylinder has an apparently uniform internal diameter of 100 Å. The external diameter of the annulus is 200 Å. The classical complement ‘rings’ visualized on membranes after complement lysis represent such C5b‐9 cylinders perpendicularly oriented on the membranes. The thin‐walled cylinder is anchored in the membrane maim and the annulus located in the exterior membrane glycocalyx. At the sites of attachment of the C5b‐9 complexes, the continuity of the membrane bilayer is disturbed and the presence of trans‐membrane pores is indicated The data essentially support the ‘doughnut’ theory of complement lysis.
Cytotoxic T Cells in the Lungs of Mice Infected with an Influenza A Virus
Authors: YAP K. L. , ADA G. L.
Published: June 2006

Cytotoxic T cells are present in the lungs and the bronchoalveolar washings of mice infected intravenously (i.v.) or intranasally (i.n.) with live influenza A/WSN virus. After i.v. injection, cytotoxic T cell activity in both spleens and lungs reaches a peak at 6 days when the level of infectious virus recovered from the lungs falls sharply and the mice do not die. If a lethal dose of virus is given intranasally, very high levels of virus appear rapidly in the lungs, and the development of lung consolidation follows slightly behind the appearance of cytotoxic T cells there When a non‐lethal dose of virus is given intranasally, lower levels of virus are found in the lung and the appearance of cytotoxic T cells is delayed. These results suggest that the cytotoxic T cells play a protective role if the level of virus in the lungs does not reach very high levels. After injection of antithymocyte serum, the subsequent level of cytotoxic T cell activity in the lungs was greatly reduced, suggesting that the T cells recovered in lungs had at an earlier stage been circulating cells. However, splenectomized mice develop high levels of cytotoxic T cell activity, after intranasal infection of mice, indicating that the spleen did not contribute substantially to the T cells recovered in the lungs.
Role of the Thymus in Generation of Lymphocyte Functions
Authors: JUHLIN R. , TUFVESON G. , ALM G. V.
Published: June 2006

Thymuses of 14‐day‐old mouse embryos could be maintained in organ culture, using serum‐free medium, with generation of lymphocytes that showed strong proliferate reactivity to allogeneic spleen cells in mixed lymphocyte cultures (MLC). Such organ culture derived thymocytes also developed cytotoxic activity against target cells tarrying the alloantigens of the stimulating spleen cells in the MLC. The generated cytotoxic activity was, however, proportionately lower than that seen with thymocytes of young mice. A time course study showed that cells with apparent reactivity to both allogeneic spleen cells and to the mitogen Concanavalin A (Con A) first appeared in the thymus organ cultures on day 8–9.
Irreversible Immunological Tolerance to Thymus‐independent Antigens is Restricted to the Clone of B Cells having both Ig and PBA Receptors for the Tolerogen
Authors: FERNANDEZ C. , MÖLLER G.
Published: June 2006

Mice were tolerized to the α1–6 epitope of native dextran. When their spleen cells wore removed and activated by LPS, they did not synthesize anybodies against the tolerogen. However, when cells from tolerant mice were treated with dextranase or left untreated in culture for 24 h they were activated by LPS to the synthesis of antibodies against the tolerogen When 24 h tolerized lymphocytes were treated with dextranase and transferred with immunogenic doses of dextran to irradiated mice they failed to produce antibodies against the tolerogen. In contrast, cells incubated with dextran for 2 h and thereafter dextranase treated were readily immunized by dextran in the same system It is concluded that only the B cell clones having both I preceptors and PBA receptors for the tolerogen become irreversibly tolerized, whereas B cells having Ig receptors for a different PBA are not tolerized, but remain in a resting state, even though their Ig receptors have bound the tolerogen.
Rat Memory T Lymphocytes: in Vitro Proliferation Induced by Antigens of Actinomyces viscosus
Authors: BURCKHARDT J. J.
Published: June 2006

Peripheral T lymphocytes from RIC‐Sprague‐Dawley rats primed in vivo several months earlier with Actinomyces viscosus Ny 1 reacted with a strong proliferative response upon antigenic challenge in vitro. Two different antigen preparations from A. viscosus Ny 1, a broken cell supernatant (BCS) and an extracellular fraction (EXC) gave maximum responses as measured by the uptake of [3H] thymidine at a concentration of 10 μg at day 4 of culture. Phytohaemagglutinin (PHA) used as a control provoked a maximum proliferation in the same order of magnitude. T lymphocytes from unprimed. germfree and conventional animals showed a similar stimulation to PHA, but only a marginal reaction to BCS, and were not at all activated by EXC. Evidence is thus presented for efficient removal of cells responding non‐specifically to B‐cell mitogens.
Reactivity of a Rabbit Antiserum against Highly Purified HLA‐DR Antigens
Authors: KLARESKOG L. , TRÄGÅRDH L. , LINDBLOM J. B. , PETERSON P. A.
Published: June 2006

A rabbit antiserum has been raised against highly purified, detergent‐Solubilized HLA‐DR antigens. Externally or Internally labelled surface glycoprotein from B‐lymphocytes, epidermis, macrophages, and B‐lymphoma cell lines reacted with the antiserum which precipitated polypeptide chains with the molecular weights 28.000 and 34.000 Such polypeptide chains were not observed when the antiserum was reacted with T‐lymphocytes, T‐cell lymphoma lines, kidney, brain. thymus and spermatozoa. Fab‐fragments of the antiserum completely abolished the cytotoxic action of HLA‐DR alloantisera but had no effect on the cytotoxicity mediated by HLA‐A, B and C loci antisera Moreover, the rabbit antiserum reacted with the same molecules as the HLA‐DR alloantisera. Fab‐fragments of the rabbit antiserum completely abolished the MLR response and from the kinetics of the inhibition it is concluded that the Fab‐fragments may interfere primarily with the initial recognition phase of the MLR.
Evidence for Two Types of Non‐specific Suppressor Cells Activated by the Graft‐versus‐Host Reaction in Mice
Authors: PARTHENAIS E. , LAPP W. S.
Published: June 2006

In vitro experiments were carried out to investigate the immunosuppressive effect of different populations of spleen cells obtained from animals experiencing a graft‐versus‐host reaction (GVHR). To induce the GVHR, parental lymphoid tells were injected into adult F1 hybrid mice. GVHR‐activated spleen cells (GVH‐SC) taken at different limes post‐GVHR induction were separated into adherent and non‐adherent fractions and treated with anti‐θ serum plus complement. The different types of GVH‐SC were added to either parental (donor‐type) or F1 (host‐type) normal spleen cells and cultured with sheep erythrocytes in a modified Marbrook chamber. It was found that both adherent and non‐adherent 10‐day GVH‐SC significantly inhibited the plaque‐forming cell response of F1 normal spleen cells hut not that of parental normal spleen cells. Significant suppression of the parental response was observed following the addition of 15‐day GVH‐SC or anti‐θ treated adherent and non‐adherent 7‐day GVH‐SC The results suggest that the non‐specific immunosuppressive effect of GVH‐SC is media led by GVHR‐activated macrophages and B lymphocytes found in the anti‐θ treated adherent and non‐adherent fractions of the GVH‐SC suspensions respectively.
Activation of Primed Rabbit Blood Lymphocytes by Antigen: Early Phase of Triggering and the Specificity of the Response
Authors: PAWLITA M. , ANDERSEN V. , LEFKOVITS I. , BRAUN D. G.
Published: June 2006

The early phase of antigen‐dependent triggering of rabbit blood lymphocytes (PBL) is described. Specific activation of PBL by Streptococcal vaccines requires primed lymphocytes from high responder rabbits. B‐cell memory is conferred by Ig receptors of both μ and γ chains. Antigen‐induced incorporation of leucine and thymidine reach peak values al day 5, with stimulation indices of 10 to > 100, closely followed by increases in the number of living cells. A second and less intensive phase of cell proliferation is seen on days 8–11. A large fraction of cells (between 3 and 33%) participates in proliferation. A rough calculation of the average doubling lime of cells during the exponential growth phase gives values of 8–16 h.
Natural and Immune Antibodies to Rabbit Erythrocyte Antigens
Authors: TÖNDER O. , LARSEN B. , AARSKOG D. , HANEBERG B.
Published: June 2006

Natural agglutinins to rabbit erythrocytes were found in all human sera studied. In the newborn, the antibodies were of IgG class; in the 6‐month‐old infants they were mainly of IgM class. Older children and adults had both IgG and IgM antibodies. Agglutinins to rabbit erythrocytes were also found in serum from fourteen of fifteen other species studied. The trichloroacetic acid extract from rabbit erythrocyte stromata (TCA‐preparation) contains at least three different antigenic determinants: one which we hitherto have found only on rabbit erythrocytes, one which is closely related to human blood group B antigen, and one which is closely related to the I antigen. The TCA‐preparation did not elicit delayed hypersensitivity skin reactions in humans in spite of high titred agglutinins in serum, but did so in immunized guinea‐pigs.
D‐Penicillamine and Macrophages: Modulation of Lymphocyte Transformation by Concanavalin A
Authors: BINDERUP L. , BRAMM E. , ARRIGONI‐MARTELLI E.
Published: June 2006

Pretreatment with d‐Penicillamine of rat peritoneal macrophages markedly influences their ability to modulate Concanavalin A stimulated [3H]thymidine incorporation in non‐adherent rat lymph node cells. Untreated macrophages added in high numbers to the lymphocytes (ratio lymphocytes/macrophage 7.5:1) depress their [3H]thymidine incorporation. Pretreatment with d‐Penicillamine further reduces this response. Lower proportions of untreated macrophages (ratio 15:1) exert a stimulatory effect on the lymphocytes. This response is reversed by pretreatment of macrophages with d‐Penicillamine. Addition of untreated macrophages in very low proportions (40:1 and 75:1) is without effect on the [3H]thymidine incorporation by the lymphocytes. This response is markedly stimulated by pretreatment of the macrophages with d‐Penicillamine. The culture of macrophages in the presence of D‐Penicillamine increases their incorporation of [3H]d‐glucosamine, but it is without effect on their incorporation of [3H]l‐leucine. It is suggested that d‐Penicillamine enhances the functional activity of the macrophage in the regulation of the lymphocyte response to mitogens.
The Affinity of Soluble Immune Complexes for Concanavalin A
Authors: HEIMER R. , KLEIN G.
Published: June 2006

Con A‐Sepharose affinity chromatography may be used in the analysis and classification of immune complexes. Experiments with model immune complexes suggest that the degree of affinity of an immune complex for Con A‐Sepharose is determined by the antigen rather than the IgG antibody of the complex. It is possible that partial characterization of unknown antigens linked to IgG in immune complexes may be achieved in many diseases. Preliminary explorations with selected human sera indicate that the IgG containing immune complexes in Burkitt's lymphoma and nasopharyngeal carcinoma have affinity for Con A‐Sepharose. By contrast IgG containing immune complexes in chronic hepatitis B seem to lack affinity for Con A‐Sepharose.
Primate HLA Antibodies are Specifically Mitogenic for Human Lymphocytes and Act Synergistically with β2m Antibodies
Authors: RINGDÉN O. , MÖLLER E. , SANDERSON A. R.
Published: June 2006

A monkey antiserum against HLA‐A28 was raised by immunization with papain‐solubilized antigens. The titre of this serum was 1/370 for β2‐microglobulin (β2m), 1/750 for A28 and 1/200 for the cross‐reacting HLA‐A2 determinant. This serum was highly mitogenic for human blood lymphocytes. Mitogenicity was reduced or abolished after absorption of the serum on a β2m‐Sepharose column. Column‐eluted anti‐β2m antibodies were not mitogenic. However, mitogenicity was restored by reconstitution with unabsorbed and absorbed antibodies. Mitogenicity was target cell specific. Thus, cells from all individuals were activated by the unabsorbed serum. The absorbed anti‐A28 serum was mitogenic only for target cells carrying A28 and/or the cross‐reacting determinant A2, but not for cells lacking either of these antigens. A definite gene‐dose effect was recorded, since target cells homozygous for A2 responded better to polyclonal activation by the primate serum than cells which were heterozygous.
Effect of in Vivo Exposure to Allogeneic Cells upon Subsequent in Vitro T Cell Responses and upon Allograft Rejection
Authors: COOLEY M. A.
Published: June 2006

The effect of intravenous injection of 108 BALB/c spleen cells into C57B1/6J recipients was assayed by both mixed lymphocyte culture (MLC) of recipient lymphocytes, and by grafting donor (BALB/c) thyroids into recipient mice. It was observed that a single intravenous injection produced depression of proliferative and cytotoxic T cell responses in MLC of spleen, lymph node and peripheral blood lymphocytes of the recipients. This effect was specific for the sensitizing genotype (MLC of recipient und third‐party C8A/H lymphocytes was unaffected), and persisted for several days after sensitization. The pattern of this diminished response suggested that the effect was due to a combination of recruitment of reactive lymphocytes from peripheral lymphoid populations, and the generation of alloantigen (H‐2?)‐specific suppressor T cells in the spleen. In contrast to these findings in vitro, a similar sensitization led only to accelerated rejection of thyroid allgrafts in vivo.
Complement Receptors in Pathological Human Renal Glomeruli
Authors: MATRE R. , IVERSEN B. M. , THUNOLD S. , TØNDER O.
Published: June 2006

Glomerular complement receptors (GCR) were demonstrated by the adsorption of C3 coated sheep erythrocytes to cryostat sections of human kidneys. An epithelial localization of the GCR is implied by the binding pattern of the indicator cells. The GCR activity was reduced in all glomeruli where in vivo deposition of immunoglobulin and C3 along the capillary wall was observed. In fourteen out of nineteen biopsies where deposits in the mesangium were seen, reduced GCR activity was also observed, whereas GCR activity was normal in the other five. Immunofluorescence and haemadsorption tests performed on the same section showed that C3 deposits corresponded to areas showing reduced complement receptor activity. Areas outside the C3 deposits adsorbed indicator cells coated with various amounts of C3 similarly to the adsorption to glomeruli seen in normal adult and fetal kidneys. Sclerotic glomeruli showed no GCR activity. The results indicate that the complement receptors play a role for the binding of complement‐containing complexes in glomeruli.
Stimulating Capacity of Human Lymphoid Cell Subpopulations in Mixed Lymphocyte Cultures
Authors: ALBRECHTSEN D. , LIED M.
Published: June 2006

The content and yield of B cells, T cells and monocytes were investigated in cell fractions separated from human peripheral blood by various techniques. These cell fractions, which were highly enriched for either B cells, T cells or monocytes, were tested for their capacity to stimulate allogenic peripheral blood mononuclear cells in mixed lymphocyte cultures (MLC). B cells were found to be strongly stimulatory. Monocytes stimulated strongly at low stimulator responder cell ratios, while less stimulation was observed using larger amounts of cells. T cells were demonstrated to stimulate weakly, if at all.
Identification of H‐2 and Ia‐Antigen Analogues in Several Species by Immunological Crossreactions of Xenoantisera
Authors: KVIST S. , KLARESKOG L. , PETERSON P. A.
Published: June 2006

Rabbit antisera against H‐2K and D antigens react with molecules composed of 12,000 and 45,000 dalton subunits derived from human, monkey, rat, guinea‐pig, cow und pig lymphocytes. The antisera failed, however, to react with similar type molecules from chicken. A rabbit antihuman β2‐microglobulin serum reacted with chicken molecules comprising 12,000 and 45,000 dalton polypeptide chains. An antiserum against HLA‐DR antigens reacted with Ia‐antigen‐like molecules from monkey, rat, mouse, guinea‐pig, cow, pig and chicken lymphocytes. All Ia‐antigen‐like molecules displayed two dissimilar subunits. The present data suggest that xenoantisera directed against highly purified MHC antigens from one species may be useful tools in elucidating the structure of similar antigens in other species where alloantisera are not available.
Altered Proportion of Tμ and Tγ‐Cell Subpopulations in Patients with Hodgkin's Disease
Authors: ROMAGNANI S. , MAGGI E. , BIAGIOTTI R. , GIUDIZI M. G. , AMADORI A. , RICCI M.
Published: June 2006

The ability of peripheral blood lymphocytes from patients with Hodgkin's disease (HD) to form rosettes with ox red blood cells (ORBC) sensitized by anti‐ORBC purified rabbit IgM and IgG was investigated. The mean percentage of cells capable of forming rosettes with ORBC coated with IgM (EAIgM‐RFC) in the peripheral blood of either untreated or X‐ray‐treated patients with HD was significantly lower than that of normal individuals. In the same groups of patients with HD the mean percentage of T lymphocytes equipped with receptor for IgG (Tγ lymphocytes), evaluated by a mixed fluorescent rosette assay, was significantly higher than in normal controls. These data suggest that the altered proportion between Tμ‐ and Tγ‐cell subpopulations in patients with HD probably represents a disease‐related phenomenon.
Lymphocyte Multiplication in Vitro Induced by Mitogens and Antigens
Authors: HANEBERG B. , WESENBERG F. , AARSKOG D.
Published: June 2006

A simple technique requiring only 0.2 ml whole blood for measuring the response of lymphocytes in cultures to each of various mitogens and antigens has been elaborated. The response is quantified by comparing the number of lymphocytes with and without a stimulating agent. The increment of cell numbers is given by a cell multiplication index. In healthy subjects PHA induced almost a doubling of the cell numbers in 3 days, i.e. an index of 1.90±0.38. After 7 days the indexes for PHA, PWM and Con A were 7.25±4.12, 2.724±0.65 and 1.81±0.31, respectively. PPD and Candida‐extract induced cell multiplication in skin positive individuals, with indexes ranging from 1.12 to 3.05. In contrast, patients with various severe immune deficiencies showed decreased responses to at least one mitogen, depending on the type of the deficiency. Likewise, skin test negative individuals had no or faint in vitro response to the antigens. The method, which correlated well with the response by a conventional method for incorporation of tritiated thymidine, has a high degree of precision and sensitivity, and should lie applicable for routine use.
Possible Detection of HLA‐DR Alloantigenic Specificities in Man with Unabsorbed Rabbit Antisera
Authors: SOLHEIM B. G. , FUKS A. , SMITH L. , STROMINGER J. L. , THORSBY E.
Published: June 2006

Thirty‐six rabbits were immunized with precipitin lines formed between detergent solubilized membrane fractions from HLA‐D‐homozygous cells and a rabbit antiserum to human B cells (anti‐p 23/30). Thirty‐one of the rabbits produced B‐cell specific cytotoxic antibodies and twelve of the antibodies were also precipitating in gel diffusion. In cytotoxicity tests six of the antisera showed clear preferential reactivity with the immunizing HLA‐D (DR) antigen. Thus immunizazation of rabbits might prove valuable for the production of HLA‐DR typing reagents.
Immunological Tolerance to the Thymus‐Independent Antigen Dextran can be Abrogated by Thymus‐Dependent Dextran Conjugates: Evidence against Clonal Deletion as the Mechanism of Tolerance Induction
Authors: MÖLLER G. , FERNANDEZ C.
Published: June 2006

Tolerance to the α1–6 epitope of native dextran B512 was found to be very stable and could not be broken by the injection of dextran conjugated to several substances, such as protein A, keyhole limpet haemocyanin, edistin, concanvalin A or Staphylococcus bacteria, strain Cowan. However, when tolerant mice were injected with dextranase, all the above conjugates induced a strong anti‐α1–6 immune response. In contrast, native dextran itself never induced a response in tolerant, dextranase‐treated mice. It was concluded that tolerance only affects the specific B‐cell subpopulation that can respond to the polyclonal B‐cell‐activating (PBA) property of dextran, whereas other specific B cells having PBA receptors for, e.g., signals delivered by collaborating T cells remain in a resting state. These B cells can respond in a specific immune response against the tolerogen after removal of the antigen, which blocks the Ig receptors and therefore prevents them from passively focusing the antigen. Thus, immunological tolerance is not caused by clonal elimination of the antigen‐specific clone, but only affects a small subfraction of cells with Ig receptors against the tolerogen.
Human Leucocyte Migration Inhibitory Factor (LIF)
Authors: BENDTZEN K.
Published: June 2006

The cyclic nucleotides guanosine 3′,5′‐cyclic monophosphoric acid (3′,5′‐cGMP) and cytidine 2′,3′‐cyclic monophosphoric acid (2′,3′‐cCMP) but not cyclic phosphodiesters derived from the bases adenine and uracil preserved LIF activity against the blocking effect of the serine protease inhibitor phenylmethylsulphonyl fluoride (PMSF). Phosphomonoesters derived from guanosine and cytidine as well as 2′,3′‐cGMP and 3′,5′‐cCMP were all inactive, indicating specificity for phosphodiester bonds and their respective positions in the two active nucleotides. The protection afforded by 3′,5′‐cGMP and 2′,3′‐cCMP was dose dependent. Thus, using 10−3m PMSF, 3,5′‐cGMP was active at concentrations higher than 10‐5 to 10‐4m, and 2′,3‐cCMP at concentrations higher than 3 × 10−4 to 10−3m. The more pronounced LIF‐inhibitory effect obtained by increased concentrations of PMSF could be overcome by raising the levels of the nucleotides, indicating that the interactions between PMSF and the nucleolides with LIF were mutually exclusive. The possibility that 3′,5′‐cGMP and perhaps 2′,3′‐cCMP function as modulators of LIF is discussed, and models for the function of this lymphokine arc proposed.
HLA‐D Restriction of the Macrophage‐Dependent Response of Immune Human T Lymphocytes to PPD in Vitro: Inhibition by Anti‐HLA‐DR Antisera
Authors: BERGHOLTZ B. O. , THORSBY E.
Published: June 2006

The response of T cells from sensitized individuals to low doses of PPD in vitro is macrophage‐dependent. By testing different allogeneic combinations of macrophages and T lymphocytes, it was found that an optimal response required that antigen be presented by macrophages sharing at least one of the HLA‐D determinants of the T cell donor. Antisera recognizing HLA‐A, ‐B or ‐DR antigens were found to be able to inhibit this proliferative response. The anti‐HLA‐DR antisera were found to exert their inhibitory effect only when directed towards an antigen shared by the donors of the T lymphocytes and the macrophages. Anti‐HLA‐A and ‐B sera, however, had an inhibitory effect when reactive with the responding T lymphocytes, irrespective of their reactivity with the cooperating macrophages. It is concluded that an optimal secondary response of in‐vivo‐immunized T lymphocytes to PPD requires the combined recognition of the antigen and ‘self’ membrane structures encoded by the HLA‐D locus.
Effect of Cyclic Nucleotides, Isoproterenol and Cholera Toxin on DNA Synthesis Triggered by Mitogens
Authors: EMIN M. , DUBISKI S. , CINADER B.
Published: June 2006

The effect of cyclic nucleotides, isoproterenol and cholera toxin on phytohaemagglutinin (PHA), concanavalin A (Con A) and anti‐allotype‐induced rabbit lymphoid cell proliferation was examined. Cholera toxin in concentrations ranging from 10−8μg to 1 μg per culture inhibited DNA synthesis, triggered by PHA, Con A and nocardia water‐soluble mitogen (NWSM). It had the opposite effect on stimulation with antibodies directed against allotypic specificities of the immunoglobulin light chains: over the entire range of tested concentrations, cholera toxin stimulated DNA synthesis triggered by antibodies to Ab4, Ab5, Ab9 and to Aa1 allotypic specificities. Relatively high concentrations of dibutyryl adenosine 3′:5′‐cyclic monophosphate (cAMP) and isoproterenol (10−3m) inhibited mitogen‐stimulated thymidine incorporation; lower concentrations (10−9m) had an enhancing effect. A similar enhancing effect was observed when high (10−3m) concentrations of dibutyryl guanosine 3′:5′‐cyclic monophosphate (cGMP) were used.
The Placental Fcγ Receptors Studied Using Immune Complexes of Peroxidase
Authors: MATRE R. , HAUGEN Å.
Published: June 2006

Immune complexes of horseradish peroxidase CHRP) and rabbit IgG antibodies 10 HRP were used to study the Fey receptors in normal human placenta. Cryostat sections of placental tissue were incubated with the complexes, and the peroxidase activity was revealed histochemically. The bound complexes were localized to the apical surface of the trophoblasts and endothelial cells of the fetal stem vessels. Binding also occurred within the wall of some fetal vessels, to stromal cells and occasionally to areas corresponding to (he trophoblastic basement membrane. The strongest binding was obtained with immune complexes prepared at slight antigen excess, Eight‐ to sixteen‐fold increased concentration of human and rabbit IgG was needed lo block the binding of immune complexes. Bovine and porcine IgG did not block the binding. Treatment of tissue sections with neuraminidase enhanced the binding activity of the receptors. The technique is very convenient for studies of Fey receptors in tissue. However, unfixed frozen placental tissue was run suitable for ultrastructural studies.
Late Appearance of an IgA(κ) Monoclonal Protein in a Patient with IgG(κ) Multiple Myeloma: Sharing of Idiotypic Specificities between the Two Serum Proteins
Authors: SCOLARI L. , VAERMAN J.‐P. , CASTIGLI E. , VOLIANI D. , SALSANO F. , MASALA C. , DI GUGLIELMO R.
Published: June 2006

A patient with an IgG(χ) monoclonal serum protein developed in the course of the disease a second monoclonal spike of the same light chain type and of the IgA class. The latter monoclonal protein progressively increased and eventually exceeded the first IgG(χ) protein. Antigenic analysis of the two myeloma proteins demonstrated that they shared idiotypic determinants. Immunofluorescence studies, carried out with anti γ and anti α reagents lagged with different fluorochromes revealed that the two isotypes were produced by different plasma cells. The data are discussed in the prospect of a possible transitional mechanism from IgG to IgA synthesis within a single B cell clone.
Lymphocyte Subpopulations in Man: No Selective Loss during Isolation from Defibrinated Blood
Authors: JOHNSEN H. E. , MADSEN M.
Published: June 2006

Human lymphocytes isolated from defibrinated blood are characterized especially in relation to the total yield of lymphocytes and their subpopulations identified by membrane markers. The defibrination per se gives a significant loss of monocytes and granulocytes but no loss of lymphocytes, indicating that no selective loss of lymphocytes occurs. In a comparison of heparin‐stabilized and defibrinated blood no difference in yield is found during every single step of the isolation procedure. Quantitation of E‐RFC and Smlg‐positive lymphocytes gives no differences in comparing the respective isolated suspensions of mononuclear cells. The observed difference in EA‐ and EAC‐RFC can freely be ascribed to the difference in monocyte contamination. Further, no correlation is found between total lymphocyte yield and the relative number of the subpopulations identified, i. e. E‐RFC, Smlg‐positive, Fc‐receptor‐ and complement‐receptor‐bearing lymphocytes. Hence it is concluded that defibrinated blood is optimal as blood source for the isolation of mononuclear cells when lymphocyte subpopulations are studied and enumerated.
Lymphocyte Subpopulations in Man: Ox Erythrocytes as Indicators in the EA‐ and EAC‐Rosette Tests: Serological and Technical Aspects
Authors: JOHNSEN H. E. , MADSEN M.
Published: June 2006

Serological properties of ox erythrocytes (ORBC) make it possible to select cells which exhibit weak agglutinability despite strong antibody sensitization. This property and the non‐binding of unsensitized ORBC to lymphocyte surface membranes make these cells excellently suited as indicators in techniques for the identification of erythrocyte‐antibody (EA) and erythrocyte‐antibody‐Complement (EAC) rosette‐forming lymphocytes (RFC). This report describes the relevant serology for the selection of appropriate cells and antisera. Further, some of the technical aspects of these tests are discussed. A simple method for the sensitization of ORBC with complement is described. The basis for this method is the naturally occurring complement‐binding anti‐ORBC antibodies of the IgM class in human sera. After zymosan treatment the sera are deficient in the fifth component of complement and hence non‐haemolytic, which make these sufficient as sensitizing agents in die preparation of EAC indicator cells. The relations of the FA‐and EAC‐RFC to be established T and B lymphocyte subpopulations are revealed by the enrichment and depletion of lymphocytes rosetting with 2‐aminoethylisothiouronium bromide (AET)‐treated sheep erythrocytes (SRBC).
Non‐specific Acid Esterase Activity in Rat Lymphocytes
Authors: Fossum S.
Published: June 2006

In a first night's collection of rat thoracic duct lymphocytes B‐ and T‐cells were distinguished on cytocentrifuge smears by complementary markers: B‐cells by rosette‐formation withStaphylococcus aureusStrain Cowan 1 armed with rabbit antiserum to rat F(ab')2and T‐cells by high uptake of3H‐uridine demonstrated by autoradiography. On incubation of the cytocentrifuge smears for demonstration of alpha‐naphthyl‐acetate esterase (ANAE)‐activity, most B‐cells showed large, intense colour deposits often located in the uropod. T‐cell ANAE‐activity on the other hand. was variable, possibly due to variations in experimental conditions. When present, the T‐cell colour deposits were small, dot‐like and of low intensity. ANAE‐activity may then he used to differentiate between rat T‐ and B‐cells, being a marker for the latter cell type. This is unexpected as others have reported that ANAE‐activity is specific for T‐cells in mouse and man. Studies with inhibitors indicate that the rat B‐cell enzyme belongs to the acetylesterases.
Many Bacterial Species are Mitogenic for Human Blood B Lymphocytes
Authors: BANCK G. , FORSGREN A.
Published: June 2006

Thirty bacterial species were tested for their ability to stimulate to increased DNA synthesis in human blood lymphocytes. A definite stimulation was obtained with eighteen bacterial species. For three of these species ten different strains of each were tested, and all increased DNA synthesis. The maximum response was after 3–4 days of culture, suggesting a mitogenic effect. This was confirmed by the induction of polyclonal antibody production shown by a plaque assay, which was positive for nine of eleven species tested. Most bacterial species increased the DNA synthesis in B‐lymphocyte‐enriched and unseparated lymphocytes but had negligible activity on T‐lymphocyte‐enriched cultures. Among bacteria with a mitogenic effect and ability to induce polyclonal antibody production are Staphylococcus aureus strain Cowan I with a high content of protein A and many common human pathogens such as Haemophilus influenzae, Mycobacterium tuberculosis, Neisseria gonorrhoeae. Streptococcus group A and Streptococcus pneumoniae.
Lymphocyte Associated β2‐Microglobulin Studied by Crossed Radioimmunoelectrophoresis
Authors: PLESNER T.
Published: June 2006

A sensitive radioimmunoelectrophoretic assay was employed in the study of β2‐microglobulin (β2) from human blood lymphocytes, allowing the detection of 2.5 pmol β2m corresponding to 3 × 106 lymphocytes. The intrinsic (amphiphilic) membrane proteins were solubilized and examined in crossed immunoelectrophoresis against rabbit antisera in the presence of non‐ionic detergent. The β2m associated precipitate was traced by post‐electrophoretic incubation with 125I‐labelled antibodies to β2m and autoradiography, A polyspecific rabbit antiserum to human lymphoid cells retained its capacity to precipitate lymphocyte β2m after absorption with isolated (on an immunosorbent column), showing that lymphocyte β2m is complexed to other molecules. No free β2m was found on lymphocytes when examined against the absorbed anti‐serum to human lymphoid cells in an intermediate gel and anti‐β2m in a reference gel.
Alloantigen‐Induced Enhancement and Suppression of Human Cytotoxic Lymphocyte Responses to Autologous Lymphoblastoid Cell Lines
Authors: ZARLING J. M. , BACH F. H.
Published: June 2006

In vitro sensitization of human lymphocytes to autologous lymphoblastoid cell lines (LCL) has been shown to give rise to cytotoxic lymphocytes capable of lysing autologous as well as allogeneic LCL cells. However, allogeneic LCL cells were found to be markedly less effective than autologous LCL cells in terms of generating lymphocytes capable of lysing autologous LCL cells. The addition of allogeneic LCL cells or allogeneic normal lymphocytes to a mixture of responding lymphocytes and X‐irradiated autologous LCL cells suppressed the generation of cytotoxic lymphocytes against autologous LCL cells. Furthermore, suppressor T cells generated in allogeneic mixed leucocyte culture (MLC) and supernatants from MLC likewise decreased the generation of cytotoxic lymphocytes to X‐irradiated autologous LCL cells. In contrast to the findings that alloantigens suppress the generation of cytotoxicity to X‐irradiated autologous LCL cells, which ordinarily induce strong cytotoxic responses, were the findings that allogeneic stimulating cells and supernatants from MLC enhanced cytotoxic responses to autologous ultraviolet light or extensively heat‐treated LCL cells that induce weaker cytotoxic responses. The possible mechanisms whereby alloantigens enhance or suppress cytotoxic responses to autologous abnormal cells and the implications of these findings are discussed.
Gut‐Associated IgA Deficiency in Lepromatous Leprosy
Authors: SAHA K. , AGARWAL S. K. , MISRA R. C.
Published: June 2006

Sera, intestinal secretions and intestinal biopsies were collected from twelve biopsy‐proved lepromatous leprosy patients with the help of a capsule invented by Roy Choudhury. Sera from another twenty‐five proved lepromatous cases were also included. Sera and intestinal aspirates from twenty‐five normal subjects and twenty patients wish intestinal tuberculosis were also taken as controls. Acid‐fast organisms, morphologically resembling Mycobacterium leprae, were detected in the intestinal aspirates of only two leprosy patients. Immunoglobulin levels in their sera and elevation secretions were estimated by the single radial immunodiffusion technique. Significant elevation of serum IgG, IgA and IgM. selective IgA deficiency in intestinal aspirates and flattening of intestinal villi along with mononuclear cell infiltration were conspicuous observations in the leprosy group. On the other hand, the patients with intestinal tuberculosis showed elevation of the IgG level in serum as well as in the intestinal secretions. It was postulated that persistent challenge by M. leprae or its antigens to the IgA immunocytes of the intestinal epithelium might have induced tolerance leading to IgA deficiency and subsequent subtotal atrophy of the intestinal villi in the patients with lepromatous leprosy.
Fc Receptor Bearing ‘Hairy Cells’ of Leukaemic Reticuloendotheliosis Bind Soluble Antigen‐Antibody Complexes and Adhere to Immobilized Complexes, but Fail to Mediate Antibody Dependent Cellular Cytotoxicity
Authors: ALEXANDER E. , SANDERS S.
Published: June 2006

The majority of hairy cells from three patients with leukaemic reticuloendotheliosis were adherent cells bearing surface immunoglobulin, complement receptors, and Fc receptors. Highly purified populations of malignant hairy cells, which readily bound soluble antigen ‐antibody complexes in suspension and were able to adhere to immobilized antigen‐antibody complexes, were examined for their ability to mediate ADCC.* Two patients with greater than 90% FcR positive cells failed to mediate ADCC. When initially examined, a third paitent, with fewer malignant cells, demonstrated a less marked impairment of cytolysis. When reexamined at a later date, this patient had an increased number of hairy cells and concomitantly demonstrated more impaired ADCC effector cell activity. Absorption of surface immunoglobulin bearing cells onto plastic surfaces coated with goat anti‐human immunoglobulin resulted in a complete depletion of FcR, sIg positive, hairy cells. The remaining nonadherem fraction, containing 5.5% FcR positive, sIg negative cells, was able to mediate ADCC as effectively as the normal controls. These results indicate that although FcR bearing hairy cells readily bind soluble antigen‐antibody complexes and adhere to immobilized complexes, they were unable to mediate ADCC.
Destruction of Sensitized Erythrocytes by Human Monocytes in Vitro: Effects of Cytochalasin B, Hydrocortisone and Colchicine
Authors: FLEER A. , VAN SCHAIK M. L. J. , VON DEM BORNE A. E. G. KR. , ENGELFRIET C. P.
Published: June 2006

The purpose of this study was to characterize the destruction of sensitized erythrocytes by human blood monocytes in vitro The incubation in vitro of human monocytes with 51Cr‐lalbelled human erythrocytes sensitized with IgG rhesus alloantibodies anti‐D EAIgG anti‐D) resulted in release of51Cr from the erythrocytes (lysis) as well as uptake of 51Cr‐labelled erythrocytes by the monocytes (phagocytosis). The lysis of EAIgG anti‐D by monocytes was not dependent on phagocytosis, because cytochalasin B, which inhibited phagocytosis of EAIgG, enhanced lysis. In contrast, hydrocorlisone and colchicine inhibited lysis, but had no effect on phagocytosis. These agents did not affect binding of EAIgG anti‐D to monocytes. The effect of these agents on lysis corresponded to their effect on release of lysosomal enzymes by monocytes. The release of lysosomal enzymes, when induced by EAIgG anti‐D, was, likewise, enhanced by cytoehalasin B and inhibited by hydrocortisone and colchicine. A significant correlation was found between lysosomal enzyme release and lysis. Together, these results strongly suggest that lysosomal enzymes. released by the monocytes when incubated with anti‐D‐sensitized erythroeytes, are responsible for the cytotoxic activity of these cells towards sensitized erythrocytes. The action of these enzymes only occurs over a short range, probably at the site of attachment of the erythrocyte, becuuse only erythrocytes that were bound to the monocytes were lysed. The finding of other investigators that removal of monocytes from suspensions of human mononuclear leucocytes results in a strong reduction in the cytotoxic activity of these leucocytes towards sensitized erythrocytes in vitro. was confirmed.
B‐Lymphocyte Colony‐forming Cells in the SJL/J Mouse Thymus Graft Repopulation: Independence on Host Thymus and Graft Genotype
Authors: CLAËSSON M. H. , WARNER N. , McKENZIE I. F. C.
Published: June 2006

We have studied the development of B‐lymphocyte colony‐forming cells (BL‐CFC) in SJL and A. SW mouse thymus grafts transplanted into lethally irradiated A. SW mice protected with SJL mouse bone marrow (A. SW‐SJL hone marrow chimaeras). The chimaeric state was confirmed by Ly allotype analysis. High incidence of BL‐CFC was found in both the A. SW and the SJL thymus grafts, indicating that SJL bone marrow generates BL‐CFC, or cells capable of generating BL‐CFC, with a high affinity for homing in the thymus irrespective of the genotype of the thymus reticulum. The incidence of BL‐CFC in the SJL thymus grafts, however, was approximately 20 times higher than the incidence of BK‐CFC in the A. SW thymus grafts. Thus, the extensive infiltration of ageing SJL thymus glands by BL‐CFC demonstrated recently is caused not only by SJL bone marrow B cells with a high affinity for thymus homing but also by the specific properly of the SJL thymus environment. The extensive infiltration of syngeneic thymus grafts by BL‐CFC observed in ageing SJL mice was found to occur independent of an intact host thymus function, indicating the extrathymic origin of these cells.
Idiotypic Determinants in Alloantigen Receptors I. Role in a Leucocyte Migration Inhibition Reaction
Authors: BRAUN M. , SZTEIN M. B. , SATZ M. L. , JASNIS M. A. , SAAL F.
Published: June 2006

An anti‐idiotypic serum was prepared by injecting BALB anti‐AKR serum into (BALB × AKR)F1 mice. Pretreatment of BALB anti‐AKR immune cells with this anti‐idiotypic serum plus complement abrogated a leucocyte migration inhibition reaction (LMIR) against AKR extracts but not against purified protein derivative. By itself, the serum induced LMIR in immune cells but not in normal cells. The reaction was strain‐specific and anti‐Thy 1,2 plus complement sensitive. These results indicate that alloantigen receptors, able to trigger an LMIR in immune cells, have idiotypic determinants similar to those of circulating antibodies of the same specificity. Thus the LMIR is a good model for the study of receptors in cell‐mediated immune reactions of primed cells.
Substrate Specificity of the Human Lymphokine Leucocyte Migration‐Inhibitory Factor (LIF): Radioenzymic Assay and Inhibition by cGMP
Authors: BENDTZEN K.
Published: June 2006

The human lymphokine, leucocyte migration‐inhibitory factor (LIF), appears to be a serine esterase and protease by virtue of its susceptibility to the irreversible enzyme inhibitor, phenylmethylsulfonyl fluoride (PMSF), and by the ability of arginine esters and amides to protect LIF against PMSF‐induced inactivation. In this paper, three methods are described by which putative substrates for LIF: may be investigated. Thus, molecules satisfying the substrate specificities of this lymphokine should (1) protect LIF against inactivation by PMSK. (2) reduce LIF activity in vitro on polymorphonuclear leucocytes, and (3) reduce the esterolytic activity of purified LIF‐rich supernatants. The first two reactions were tested hy means of the leucocyte migration agarose technique; the third reaction was tested by a sensitive enzyme assay using tritiated tosyl arginine methyl ester as substrate. Guanosine 3′,5’‐cyclic monophosphoric acid, which is capable of protecting LIF against PMSF‐induced inhibition, also inhibitied the esterolytic activity of the purified LIF preparation. Four synthetic oligopeptide substrates for trypsin, thombin and plasmin were investigated. Only one, the thrombin‐ and trypsin‐specific benzoyl‐phenylalanyl‐valyl‐arginie‐p‐nitroanilide, possessed high affinity for the LIF molecule and may therefore prove to be a potent substrate for this lymphokine.