bluuebunny/crossref_metadata_2025 huggingface dataset:
The Immunosuppressive Effect of Extracorporeal Irradiation of the Blood Using a Portable 90Sr‐90Y Source and Small Transit Doses
Authors: BIRKELAND S. A.
Published: June 2006

The immunosuppressive effect of extracorporeal irradiation of the blood (ECIB) using small transit doses (TD) over a long time period was studied. The blood of 15 uremic patients undergoing hemodialysis in preparation for kidney transplantation was treated with 15‐, 25‐, or 41‐rad TDs for a total of 150 h, using the beta‐emitting radioisotope 90Sr‐90Y. The number of blood volumes (BV) irradiated ranged from 159 to 265. The irradiation resulted in selective lymphocytopenia, but the effect on the immune response of lymphocytes, as measured in vitro with lymphocyte transformation tests, varied. Six kidney transplantations were performed resulting in rejection in two cases, one which was easily reversed and one with many rejection episodes, ending in death. Thus treatment with small TDs over long periods can reduce lymphocyte counts, but any subsequent immunosuppressive effect is uncertain. Our results do not support the hypothesis that such irradiation can influence the immune response of lymphocytes without destroying the cells.
Complex Nature of the Production of ‘Effector Cells and Their Interaction with Target Cells in the M‐Antigen System of the Mouse
Authors: LILLIEHÖÖK B. , JACOBSSON H. , BLOMGREN H.
Published: June 2006

The capacity of lymphocytes from mouse strain CBA to generate ‘effector’ cells against the H‐2‐compatible, M‐antigen‐incompatible strain C3H and their interaction with such target cells were investigated. It was observed that CBA lymphocytes injected in, and 5 days later obtained from, the spleens of irradiated C3H × CBA hybrids (‘educated cells’) could strongly inhibit the growth of C3H × CBA bone marrow cells but were almost nonresponsive to C3H × C57BI bone marrow targets (H‐2‐incompatible). CBA lymphocytes educated in irradiated C3H × C57B1 hosts displayed reactivity against C3H × C57B1 and CBA × C57B1 but not against C3H × CBA bone marrow target cells. Additional tests indicated that the M antigen determined by C3H is expressed td approximately the same extent on C3H × CBA and C3H × C57B1 cells and that the M antigen on C3H does not cross‐react immunologically with antigens on C57B1 cells. Moreover, it was observed that CBA × C57B1 lymphocytes were triggered to cell proliferation by C3H antigens but were unable or had a highly reduced capacity to develop ‘effector’ cells in response to this antigenic stimulus. These results indicate that generation of ‘effector’ cells and their interaction with target cells are very complex processes.
Impact of Ig‐Anti‐Ig Column Fractionation on T‐Lymphocyte Functions
Authors: RUBIN B. , JØRGENSEN P. N. , GÜTTLER F. , HØIER‐MADSEN M.
Published: June 2006

C57B1/6 anti‐PS 15‐immune spleen cells were fractionated on immunoglobulin (Ig)‐anti‐Ig antibody columns in medium containing EDTA or in EDTA‐free medium. The antigen‐specific cytotoxic potential of immune cells in vitro was only slightly enriched after passage through the columns in the absence of EDTA, whereas immune cells passed through the columns in the presence of EDTA displayed a cytotoxic potential enriched about three times compared with unfractionated cells; this is similar to the enrichment θ‐positive cells. The lack of increase of the cytotoxic potential of immune cells after passage through the columns in the absence of EDTA was shown to be due to the adsorption of subpopulations of cytotoxic T lymphocytes. These cytotoxic cells could be doted with EDTA‐containing medium, and the findings suggest that their adsorption may be mediated via the interaction between cell‐bound Fc receptors and the column antigen‐antibody complexes.
Properties of Fcγ Receptors in Normal and Malignant Human Tissues
Authors: MATRE R. , TÖNDER O. , THUNOLD S. , SOLHAUG J. H.
Published: June 2006

Properties of the IgG receptors were studied by treating tissue sections or suspensions of peripheral mononuclear cells with various chemical reagents and determining changes in their ability to react with IgG‐sensitized erythrocytes (EA). A heterogeneity of the Fc receptors was revealed. lodoacetamide, 2‐mercaptoethanol, sodium azide, EDTA, and a pH varying from 6.0 to 8.2 had no effect on Fc receptors in sections of normal lymphoreticular tissue and malignant tissue. Formaldehyde, low pH, and high salt concentrations affected receptor activity to various degrees. Receptors in liver sections and on monocytes were generally more resistant than receptors in spleen sections and on B lymphocytes. Receptors in malignant tissue behaved either like receptors in spleen or like receptors in liver. Although all tissues were sensitive to periodic acid, the Fc receptors in malignant tissue were always more resistant.
Mixed Lymphocyte Reactivity and Cell‐Mediated Lympholysis to D‐End Differences of the Murine Major Histocompatibility Complex
Authors: HODES R. J. , SCHMITT‐VERHULST A.‐M. , HATHCOCK K. S. , SHEARER G. M.
Published: June 2006

In vitro mixed lymphocyte culture (MLC) responses and the in vitro induction of cell‐mediated lympholysis (CML) were studied in congenic strain combinations in which the responding and stimulating strains differed either at the entire major histocompatibility complex (MHC) or only at the D end of the MHC. In contrast to previously reported studies, the relative strengths of stimulation by ‘D end only’ differences or by whole MHC differences were examined by stimulating identical responding populations with titrated numbers of stimulating cells that differed from the responder either at the D end only or over the entire MHC. When tested in this manner isolated D‐end differences were sufficient to generate significant MLC and CML responses in each combination tested. Several ‘D end only’ differences (the responses of B10.A to B10.A(2R); of B10.A(2R) to B10.A; of B10.D2 to B10.HTG; and of B10.HTG to B10.D2 were several fold less efficient in stimulating MLC and CML responses than were control stimulating cells differing over the whole MHC. In contrast, when the mutant D‐end allele da was present on the stimulating cell (the responses of B10.D2 to B10.D2(M504) and of B10 to B10.D2(R106)), stimulation by M isolated D‐end difference was comparable to stimulation by broader MHC differences. These findings are discussed in terms of the possible functional complexity of the D region.
Effects of Granulocyte Neutral Proteases on Complement Components
Authors: JOHNSON U. , OHLSSON K. , OLSSON I.
Published: June 2006

The proteolytic effects of collagenase and elastase from human granulocytes were investigated on the human complement components C3 and C5 in serum and with purified components. The conversion of C3 was analyzed with crossed immunoelectrophoresis as described by Ganrot, and the conversion of C5 was detected with immunoelectrophoresis according to Scheidegger's method. Collagenase converted C3 to C3b but had no detectable effect on C5. Elastase converted C3 to C3b and converted C5 to a C5b‐like fragment. The proteolysis by collagenase and elastase in serum was not detectable until the molar ratio for enzyme to the protease inhibitors α1‐antitrypsin and α2‐macroglobulin was greater than 1.
Complement Receptors in Human Renal Glomeruli
Authors: MATRE R. , TÖNDER O.
Published: June 2006

Glomeruli in Cryostat sections of foetal and adult renal tissues adsorbed sheep erythrocytes sensitized with IgM antibody and human or mouse complement (EAC). Activated complement was essential for the reaction to occur. The receptor had specificity for the C3b fragment of C3 and is probably of protein or glycoprotein nature. Complement receptors in glomeruli may explain the deposition of complement components or complement‐containing complexes in immune‐complex glomerulonephritis.
Cell‐Mediated Cytotoxicity for Melanoma Tumor Cells: Detection by a (3H) Proline Release Assay
Authors: SAAL J. G. , RIEBER E. P. , RIETHMOLLER G.
Published: June 2006

An in vitro lymphocyte‐mediated cytotoxicity assay using [3H]proline‐labeled target cells is described. The assay, modified from an original procedure of Bean et al., assesses the release of [3H]proline by filtering the total culture fluid containing both trypsinized tumor cells and effector cells. Filtration is performed with a semiautomatic harvesting device using low suction pressure and large‐diameter glass filters. Pretreatment of filters with whole serum diminishes adsorption of cell‐free radioactive material considerably and thus increases the sensitivity of the assay. Nearly 100% of the radioactivity could be recovered with this harvesting device. The technique allowed the detection of cytolytic activities of lymphocytes after 6 h of incubation. Lymphocytes from patients with primary malignant melanoma showed a significantly higher cytolytic reactivity(P< 0.001) than normal donors’ lymphocytes against three different melanoma cell lines. In a series of parallel experiments on 36 patients and IS normal donors, this modification of the [3H]proline test was compared with three different assays: the conventional microcytotoxicity test of Takasugi and Klein, the original [3H]proline microcytotoxicity test of Bean et al., and the viability count of tumor cells.
Modification of the Response of Human T Lymphocytes to Phytomitogens by Cocultivation with Unresponsive Non‐T Leukocytes
Authors: BLOMGREN H.
Published: June 2006

Experiments were conducted to ascertain to what extent the mitogen response of human peripheral T cells is influenced by contaminated, nonresponding leukocytes, T‐ and B‐lymphocyte preparations, extensively depleted of monocytes, exhibited extremely poor responses to pokeweed mitogen and concanavalin A compared with the original unfractionated cell population. A strongly reduced phytohemagglutinin response was noted in the B‐cell but nut in the T‐cell fraction. Mixtures of cell preparations enriched in T or B cells yielded stimulation higher than expected. Mitomycin‐C‐treated T‐cell preparations mixed with untreated preparations enriched in B cells gave slightly higher responses than expected. However, highly enhanced responses were observed when untreated T cells were mixed with mitomycin‐treated cell suspensions enriched in B cells. A similar enhancing effect was noted when mitomycin‐treated cell suspensions enriched in adherent cells were added. It is concluded that the mitogen reactivity of T cells, on a cell‐for‐cell basis, may be enhanced tenfold or more by cocultivation with unresponsive non‐T leukocytes. A reduced mitogen reactivity of peripheral lymphoid cells from patients with various types of disease may thus either reflect a decreased proportion of responsive lymphocytes or unresponsive non‐T lymphocytes capable of enhancing mitogen reactivity, or both.
Differential Cytotoxicity of Activated Lymphocytes on Allogeneic and Xenogeneic Target Cells
Authors: STEJSKAL V.
Published: June 2006

A competitive inhibition assay was used to define the specificity of the in vitro cytotoxicity of activated lymphocytes. Human and mouse effector cells were produced by prestimulation with phytohemagglutinin, pokeweed mitogen, or purified protein derivative for several days. The addition of increasing numbers of unlabeled Chang cells to a fixed number of stimulated human lymphocytes and 51Cr‐labeled Chang cells gradually decreased chromium release. Admixture of unlabeled human bladder tumor cells, lung cells, or monkey kidney cells had a similar effect, whereas mouse L cells were not inhibitory. The reverse was found to be true when stimulated mouse lymphocytes and 51Cr‐labeled mouse L cells were incubated. In this case, the addition of unlabeled L or rat tumor cells effectively inhibited 51Cr release, whereas human or monkey cells were less inhibitory. Both human and mouse cells inhibited the cytotoxicity of human effector lymphocytes towards mouse L cells. The cytotoxic effects of mouse lymphocytes on human cells were also decreased by addition of unlabeled human, monkey, or mouse cells, but in this case human cells were most inhibitory. The results indicate that activated lymphocytes recognize several surface structures on the target cells. Some of these structures may be shared by cells of different species origin, whereas others seem to be unique for cells from phylogenetically closely related species.
Binding of IgM by Human T Lymphocytes
Authors: GMELIG‐MEYLING F. , VAN DER HAM M. , BALLIEUX R. E.
Published: June 2006

Ox etythrocytes, sensitized with IgM‐type rabbit antibodies, were found to form rosettes with human blood lymphocytes. Granulocytes, monocytes, and macrophages appeared to lack this property. The rosette formation could be inhibited with human IgM and its Fc fragments. IgM‐binding lymphocytes were also present in cell suspensions isolated from human tonsils and thymuses and among blood lymphocytes cultured in vitro. The outcome of combined marker tests and cell separation experiments disclosed that the resetting lymphocytes belong to the T‐lymphocyte population.
Detection of β2‐Microglobulin in Sera from Diverse Vertebrate Species by the Use of a Heterologous Radioimmunoassay
Authors: GORDON S. M. , KINDT T. J.
Published: June 2006

The strong cross‐reaction between rabbit and human β2‐microglobulins, as detected by goat antisera, has permitted the development of heterologous radioimmunoassays to detect homologues of β2‐microglabulin (β2μ) in the sera of various vertebrate species. The assay based on the inhibition of the binding reaction between goat anti‐rabbit β2μ and human β2μ divided the sera examined into two groups, one giving high levels of inhibition (∼80%), and the other lower levels (∼20%). There was no evident correlation between this division and phylogenetic classification. When the same group of sera was tested using the assay based on the inhibition of binding of rabbit β2μ to goat anti‐human β2μ, a wide range of maximal levels of inhibition was observed, with clustering of phylogenetically related species
Prozone Effects in Indirect Immunofluorescence
Authors: LINDER E. , MIETTINEN A.
Published: June 2006

A marked prozone effect was observed in indirect immunofluorescence (IFL) with rabbit antisera against rat renal proximal tubular epithelial brush border (BB) antigens: anti‐BB antibodies were not detectable if used in high concentrations but were readily demonstrated if sufficiently diluted. The prozone effect occurred in spite of demonstrated binding of anti‐BB antibodies to target antigens. No prozone was seen when direct IFL was used, and ‘cross‐reaction’ antigens in small‐intestinal epithelial brush border were detectable by indirect IFL without prozone. It was concluded that the anti‐immunoglobulin conjugates were unable to reach antigenic determinants on tightly clustered immunoglobulin molecules. Thus prozone effects seem to depend primarily on the density of antigen determinants in the target tissues.
A Mechanism of Migration Inhibition Involving Components of the Coagulation System
Authors: MEADE C. J. , LACHMANN P. J. , BINNS R. M.
Published: June 2006

Lymph from pigs given bacillus Calmette‐Guérin (BCG) and stimulated in the drainage area of a cannulated lymph node with tuberculin (PPD) had potent in vitro macrophage migration inhibitory activity. A similar observation has previously been made in sheep by other workers. In the sheep, part of, and in the pig. almost all of, this inhibitory activity was produced, not by a factor resembling the so‐called ‘migration inhibitory factor’, but by a mechanism involving a protease and fibrinogen. It is suggested that exudate cells, having potent coagulation‐promoting activity, can promote the deposition of fibrin about themselves, and thus ‘trapped’ are unable to migrate.
Maternal Immunocompetence
Authors: HARRISON M. R.
Published: June 2006

Lymphocytes from the peripheral blood, spleen, or para‐aortic lymph nodes of primigravida L rats carrying (L × BN)F1 (LBN) fetuses are fully capable of mounting graft‐versus‐host (GVH) reactions in LBN F1 recipients. The reactivity of lymphocytes from interstrain pregnant (L × BN) or intrastrain pregnant (L × L) rats, or from rats postpartum from these pregnancies, is equivalent to that of normal virgin females over a full dose‐response curve, ruling out both specific and nonspecific effects of pregnancy on the intrinsic GVH competence of the maternal thymus‐derived (T) lymphocyte. Attempts to block GVH reactivity with serum from pregnant rats were unsuccessful. In addition, when the distribution pattern of 51Cr‐labeled syngeneic and semiallogeneic lymphocytes was studied in intact primigravida mice, there was no difference between inter‐strain and intrastrain pregnant mice, and there was no evidence of immunologically specific ‘trapping’ in the para‐aortic lymph nodes draining the interstrain pregnant uterus.
Shared Idiotypic Determinants on B and T Lymphocytes Reactive Against the Same Antigenic Determinants
Authors: BINZ H. , WIGZELL H.
Published: June 2006

Normal rat lymphocyte populations house a high percentage of lymphocytes with idiotypic, antigen‐binding receptors for the major histocompatibility complex antigens of the rat. These receptors can be isolated from normal serum or lymphocyte supernatants. Idiotypic, antigen‐binding molecules released from normal Lewis lymphocytes were thus isolated using anti‐(Lewis anti‐DA) immuno‐adsorbents. Analysis by SDS polyacrylamide elertrophoresis using molecules labeled by external or internal means (125I or 3H) demonstrated that B lymphocytes produce molecules of ‘conventional’7S–8S IgM type. T‐lymphocyte‐derived molecules had a molecular weight of around 150,000 and consisted of two chains of similar size. Such single chains would succumb to proteolysis by normal serum factors to yield fragments in the size range of 30,000–40,000 daltons. All three groups of T‐cell‐derived molecules expressed both antigen‐binding and idiotypic markers. No evidence was obtained that any light chains are linked to the T‐receptor molecules. Serological analysis of the T‐cell molecules failed to prove the existence of any determinants of constant immunoglobulin type, nor did these molecules express antigenic markers of major histocompatibility complex types.
Off‐Signals in Lymphocyte Activation
Authors: MÖLLER G.
Published: June 2006

High‐dose unresponsiveness to concanavalin A affected T and B lymphocytes equally, although only T cells were activated by Con A. Thus, activation and inactivation signals art totally unrelated, the latter also lacking target cell specificity. The off‐signal caused cell death and is probably biologically unimportant.
IgA in Commercial γ‐Globulin Preparations
Authors: WADSWORTH C. , HANSON L. Å.
Published: June 2006

The presence of anti‐IgA has been related to adverse reactions to injection Oil infusion of IgA‐containing material. In this study IgA was demonstrated in all of the investigated commercial γ‐globulin preparations. In the material from a few producers it was, however, very low. By immuno‐gel filtration it could be shown that the IgA consisted of aggregates as well as a 7S component and fragments. By immunoelectrophoresis‐immunuofluorescence IgG‐IgA complexes could also be detected. The IgA aggregates could be almost Completely degraded by reduction‐alkylation. It is implied that such changed IgA in γ‐globulin preparations for injection may increase the risk of immunization against IgA.
Changes in the Surface of Virus‐Infected Cells Recognized by Cytotoxic T Cells
Authors: ADA G. L , JACKSON D. C. , BLANDEN R. V. , HLA R. THA , BOWERN N. A.
Published: June 2006

P‐815 mastocytoma cells develop susceptibility to immune T‐cell‐mediated cytolysis shortly after infection by ectromelia virus. Intracellular viral replication and late protein synthesis seem to be unnecessary events. Interference with early protein synthesis, however, inhibits the development of susceptibility to lysis. The important intracellular events necessary for subsequent cytolysis appear to occur within 1 hr of infection. Virus rendered noninfectious by ultraviolet irradiation but not by γ irradiation is able to induce these changes. By determining the minimum and essential events of the infectious process which result in T‐cell‐mediated cytolysis, the task of establishing the molecular changes occurring in the target cell surface membrane necessary for immune T‐cell recognition should be simplified.
Inhibition of In Vitro Immune Responses by a Fraction from Seminal Plasma
Authors: PRAKASH C. , COUTINHO A. , MÖLLER G.
Published: June 2006

We studied the inhibitory effects of a fraction of bovine seminal plasma on in vitro lymphocyte responses to specific and nonspecific ligands. Seminal plasma was fractionated by Sephadex G‐100 chromatography, and the inhibitory component was found to have a molecular weight of above 100,000 daltons. This fraction was devoid of lymphocytotoxity and could inhibit, in a dose‐dependent mode, both the primary antibody response of normal mouse spleen cells to the thymus‐dependent antigen sheep erythrocytes and the one‐way mixed lymphocyte reactions. Concanavalin A‐induced proliferate T‐cell responses were also inhibited by the same fraction. The inhibition of the T‐lymphocyte responses to Con A was complete at suboptimal concentrations of the mitogen and could only partially be overcome at supraoptimal concentrations. Four other fractions in seminal plasma were also studied in this particular system, three being without effect and one being stimulatory. The inhibitory fraction was also found to inhibit the polyclonal B‐cell responses induced by fetal calf serum. The importance of the present findings for the immunophysiology of reproduction are discussed.

G. Möller, Division of Immunobiology, Karolinska Institute, Wallenberglaboratory, Lilla Freskati, 104 03 Stockholm 50, Sweden
Antibodies to Surface Antigens of Lymphocytes and Lymphoblastoid Cells in Cold‐Agglutinin‐Positive Sera from Patients with Mycoplasma pneutnoniae Infection
Authors: BIBERFELD G. , BIBERFELD P. , WIGZELL H.
Published: June 2006

IgM antibodies reacting in the cold with surface antigens of normal human blood lymphocytes were demonstrated by indirect immunofluorescence (IFL) in cold‐agglutinin‐positive sera from 21 patients with Mycoplasma pneumonias (MP) infection. Twenty to fifty per cent of the lymphocytes were stained. MP sera reacted also with 75%‐100% of cells from two lymphoblastoid cell lines and one Burkitt lymphoma line and with about 80% of peripheral blood lymphocytes from one patient with chronic lymphatic leukaemia. The IFL reaction was negative with cells from a leukaemic lymphoid T‐cell line (Molt‐4). Lymphocyte fractionation experiments gave no evidence of a selective reactivity of MP sera with B or T lymphocytes of peripheral blond. Removal of cold agglutinins from the MP sera by absorption with human O erythrocytes significantly reduced the IFL reactivity with lymphocytes and lymphoblastoid cells, indicating the presence of I antigen on these cells. Furthermore, lymphoblastoid cells but not Molt lymphoid cells absorbed out most of the cold erythroagglutinins.
The Immunosuppressive Effect of Extracorporeal Irradiation of the Blood Evaluated by Blast Transformation and Membrane Immune Fluorescence Tests
Authors: BIRKELAND S. A. , MOESNER J. , WIIK A.
Published: June 2006

The immunosuppressive effect of extracorporeal irradiation of the blood was investigated by in vitro blast transformation tests on circulating lymphocytes and on lymphocytes isolated from lymph nodes and by B‐lymphocyte counts before, during, and after the treatment. Two consecutive patient series, from a controlled clinical trial of the effect of extracorporeal irradiation of the blood on the course of kidney transplantation, were studied. B‐cell depletion was found to be relatively larger than the total lymphocyte depletion, in vitro transformation tests on both circulating and lymph node lymphocytes showed large individual variations. There were no characteristic correlations between the results of these tests or with the number of circulating lymphocytes. Nor were there any characteristic changes in these tests during treatment. These results, together with previous experience, seem to contraindicate the value of irradiation of the blood as an immunosuppressive treatment.
Genetic Control of B‐Cell Responses
Authors: COUTINHO A.
Published: June 2006

The responsiveness of spleen cells from C3H/HeJ and C3H/Tif mice to lipopolysaccharide (LPS) was compared. Around 1,000‐fold higher concentration of LPS were required to minimally activate HeJ cells, as compared with Tif high‐responder cells, to both proliferation and polyclonal antibody secretion. However, HeJ cells did respond to higher LPS concentrations (100 and 1000 μg/ml). This selective pattern of responsiveness to LPS was also observed in the polyclonal responses of both strains to LPS in vivo. Furthermore, the unresponsiveness of HeJ spleen cells was found to depend on a pure B‐cell defect in the capacity to interact and/or generate triggering signals on interaction with LPS. Thus, adherent cells, thymus‐derived lymphocytes, serum factors, and other nonspecific conditions inherent in spleen cell suspensions of low‐responder mite were not responsible for suppressing a putative B‐cell response to LPS. The present findings are compatible with the possibility that the defect in C3H/HeJ B cells reflects the absence of a structure on the cell surface membrane that is functionally responsible for mediating LPS triggering.
Evidence of Oligomeric IgA in Canine Thymus Cysts
Authors: SANDHOLM M. , KAARTINEN M. , OKSANEN A.
Published: June 2006

The content of canine thymic cysts present in involuted thymuses was studied. The major component of the cyst fluid consisted of an electrophoretically heterogeneous molecule population having cross‐reactivity with IgA and a high natural antibody activity against DNP and NIP. The major IgA fraction had a sedimentation coefficient of 10S, suggesting a dimeric form. High natural antibody activity was seen also in the 12S‐16S area on ultracentrifugation, which indicates that polymers larger than dimers also occur. However, semiquantitative analysis showed that dimeric IgA is probably a major component of canine thymic cyst fluid.
Antibody Activity of Heavy and Light Chains and Recombined IgG of Human IgG Anti‐D
Authors: FØRRE Ø. , MICHAELSEN T. E. , NATVIG J. B.
Published: June 2006

Four human sera with high titers of IgG anti‐D were studied in recombination experiments with isolated heavy and light chains. The binding to the D antigen measured by hemagglutination with O,R1R2 erythrocytes was 30–60 times weaker isolated anti‐D heavy chains and with hybrids of anti‐D heavy chains and pooled light chains than with the parent anti‐D IgG. Light chains and hybrids of light chains from anti‐D IgG and pooled heavy chains showed no binding. This indicates that the heavy chains are more important in antigen binding than the light chains in this system. It was also shown that the idiotypes need both heavy and light chains of the anti‐D molecule to give a positive reaction with specific anti‐idiotypic antiserum.
μ Heavy‐Chain Disease—A Defect in Immunoglobulin Assembly Structural Studies of the χ Chain
Authors: FRANGIONE B. , FRANKLIN E. C. , PRELLI F.
Published: June 2006

μ‐chain protein GLI is a pentameric molecule with an amino‐terminal deletion comprising 130 residues. The half‐cysteine residue (position 140) which forms the H‐L disulfide bridge in normal IgM is present. Instead of being joined to the L chain, it presumably exists as an additional inter‐H‐H disulfide bridge. The χ Bence Jones protein is of normal size and present in two forms: as monomers and dimers. The carboxy‐terminal half‐cysteine of the monomer is bound to cysteine. Possible reasons for failure of assembly between μ and L chains are briefly discussed.
Reassembly of Immunoglobulin M Heavy and Light Chains In Vitro
Authors: SCHROHENLOHER R. E. , HESTER R. B.
Published: June 2006

Reduced and alleviated monoclonal IgM was fractionated into μ and light (L) chains by gel chromatography in 1N acetic acid. Equimolar mixtures of the chains formed a noncovalently bonded structure in 0.01M sodium acetate buffer, pH 4.1, that had the properties of a half subunit. The latter reassociated into a subunit‐like structure after transfer into 0.08M sodium phosphate buffer, pH 7.5. The similarity of the reconstituted IgM subunit (IgMs) to that of the native molecule was established by its physicochemical and immunochemical properties. Comparable products were obtained on reassembly of the alkylated μ and L chains from several other monoclonal IgM. The presence of active binding sites for IgG on subunits reconstituted from the chains of proteins with anti‐IgG activity further indicated correct assembly of the μ and L chains. High yields of subunit‐like products were also obtained by assembly of μ chains from one protein and L chains from another. Evidence was obtained that L chains of appropriate specificity can substitute for the homologous chain in the formation of the active site. Heterogeneous mixtures of high molecular weight products were generated from μ and L chains that were not alkylated. Reduction and alkylation demonstrated that the products represented polymers of reconstituted IgMs. Significant levels of anti‐IgG activity were detected in the polymeric IgM generated from the chains of active proteins by precipitation with aggregated IgG.
Naturally Occurring Polymers of IgA Lacking J Chain
Authors: TOMASI T. B. , CZERWINSKI D. S.
Published: June 2006

Two of twenty IgA myeloma proteins studied were found to lack J chain. Both IgA proteins contained dimers and higher polymers (trimers, tetramers, pentamers) in proportions similar to those found in most classical ‘J‐positive’ proteins. Both the ‘J‐negative’ proteins contained bound albumin and alpha‐1 anti‐trypsin (α1AT), and reduction with mercaptoethylamine caused a release of albumin and α1AT concomitant with depolymerization of the higher polymers of IgA. These proteins formed complexes with secretory component (SC) in vitro, indicating that the presence of J chain is not a requirement for SC binding.
Restriction of Human Immune Antibodies to Heavy‐Chain Variable Subgroups
Authors: NATVIG J. B. , FØRRE Ø. , MICHAELSEN T. E.
Published: June 2006

Human immune antibodies such as anti‐Rh and anti‐Kell antibodies were tested in hemagglutination and hemagglutination inhibition experiments for VH subgroup composition. A striking VH subgroup restriction was found in several of these groups of antibodies. In the majority of cases there was a restriction to one particular VH subgroup for a single antibody specificity in a given individual. In some cases there was also an overall restriction to one particular subgroup for antibodies with the same antibody specificity. This was particularly pronounced for anti‐D antibodies, which were primarily restricted to VHII, and for the anti‐Kell, which was particularly related to VHIII. Subgroup‐specific antigens for all the main VH subgroups were blocked on combination of the antibody molecule with antigen. No relation was found between VH restriction and restriction to IgG subclass, or genetic markers or χ and λ light‐chain type.
Complete Amino Acid Sequence of the Variable Domains of Two Human IgM Anti‐Gamma Globulins (Lay/Pom) with Shared Idiotypic Specificities
Authors: CAPRA J. D. , KLAPPER D. G.
Published: June 2006

On the basis of extensive shared idiotypic specificities, two human IgM anti‐gamma‐globulins (Lay/Pom) were selected for complete amino acid sequence analysis of their variable domains. Previous studies on the variable regions of the heavy chains of these proteins had shown but eight amino acid differences, only one of which was within a complementarity‐determining hypervariable region. The complete amino acid sequence of the variable regions of the light chains of these two proteins is the subject of this report. Protein Lay is a typical VχI protein with only five ‘framework’ differences when compared with protein Roy. Protein Pom is best classified as a VχII, but in the ‘framework’ there are 16 differences between it and protein Ti. Although there are extensive differences in the first hypervariable region, the second and third light‐chain hypervariable regions have an identical sequence. The finding of two identical light‐chain and two identical heavy‐chain hypervariable regions in these two proteins, which were selected on the basis of their combining specificities and their idiotypic cross‐reactions, strongly implicates hypervariable regions in the constitution of the idiotypic determinants and the antibody combining site. Additionally, the finding of identical hypervariable regions in light chains of different V‐region subgroups fulfills a prediction of the gene‐interaction concept of antibody variability.
Factor B of the Alternative Complement Pathway on Human Lymphocytes
Authors: HALBWACHS L. , LACHMANN P. J.
Published: June 2006

A factor on human lymphocytes has been identified as factor B of the alternative pathway. Lymphocytes can replace factor B in the fluid phase formation of C3 convertase with cobra venom factor (CVF). This lymphocyte activity is inhibited by specific anti‐human factor B, and it is shown by Burkitt lymphoma cell lines cultured in the absence of any factor B source. After the reaction with CVF all the C3‐converting activity is found in the cell supernatant, and the same cells can undergo several successive cycles of ‘activation’ by CVF. Factor B is distinct from the C3b receptor, and its presence could not be detected antigenically on the lymphocyte membrane. It may be secreted by the cells, but the reaction was not affected by sodium azide or cytochalasin B. No detectable factor B activity was found in the culture medium of cells grown in the absence of CVF.
Nephritic Factor: Its Structure and Function and Its Relationship to Initiating Factor of the Alternative Pathway
Authors: SCHREIBER R. D. , GÖTZE O. , MÜLLER‐EBERHARD H. J.
Published: June 2006

Nephritic factor (NF) has a molecular weight of 170,000 and is composed of two disulfide‐linked 85,000‐dalton chains. NF assembles the fluid‐phase C3 convertase from Factors B and D, C3, and magnesium by physically incorporating itself into the enzyme complex. NF exerts its stabilizing effect on the cell‐bound C3/C5 convertase, EC, by physically associating itself with this complex. On decay of the cell‐bound enzyme NF is released into the fluid phase and retains its binding and stabilizing potential. Its activity is resistant to diisopropylfluorophosphate treatment. Because NF causes agglutination of EC, it must be endowed with more than one binding site.
Reversible Activation of Proactivator (Factor B) of the Alternative Pathway Without Cleavage of the Molecule
Authors: DAY N. K. , SCHREIBER R. D. , GÖTZE O. , MÜLLER‐EBERHARD H. J.
Published: June 2006

The serum of a patient (M.C.) with chronic glomerulonephritis and renal deposits of IgA, C3, and properdin converted C3 on overnight exposure to 0°C. The cold reaction was dependent on immunoglobulin, initiating factor, Factors B and D, and magnesium but not on properdin. Factor B, the C3‐cleaving enzyme in this reaction, was used in zymogen form. After participation in this reaction, Factor B zymogen in M.C. serum could be fully activated by cobra venom factor (CVF) at 37°C. That activation without fragmentation was not due to an abnormal form of Factor B was shown by its typical cleavage on incubation of M.C. serum with CVF or C3b or after depletion of C3b inactivator. The evidence indicates that in the cold reaction only the initial C3 convertase of the alternative pathway is formed and that this enzyme is responsible for the observed C3 consumption.
Studies of Ia Antigens on Murine Peritoneal Macrophages
Authors: SCHWARTZ R. H. , DICKLER H. B. , SACHS D. H. , SCHWARTZ B. D.
Published: June 2006

Murine peritoneal cells, both induced and noninduced, were examined For Ia antigens by a variety of techniques. Complement‐mediated cytotoxicity and indirect immunofluorescence, analyzed by both visual microscopy and the fluorescence‐activated cell sorter, detected Ia antigens on the surface of an average of 8% to 15% of cells with the morphologic and functional characteristics of macrophages. Internal radioisotope labeling studies showed that these antigens were actually synthesized by the macrophages. The antigens were borne on molecules which consisted of two components with apparent molecular weights of roughly 33,000 and 25,000 daltons. At least some of these molecules existed as a two‐chain structure of 58,000 daltons linked by disulfide bonds. Although macrophage Ia antigens appeared to be structurally similar to the Ia antigens found on spleen cells, the radioisotope labeling studies indicated that the quantity of labeled Ia‐bearing molecules isolated from peritoneal macrophages was at most 1/15 that found for B lymphocytes. In addition, anti‐Ia antisera failed to inhibit the binding of heat‐aggregated immunoglobulin to the Fc receptor of macrophages. Thus, Ia antigens appear to be present at very low levels in macrophage populations, similar to the low levels of Ia antigens found in T‐lymphocyte populations. These studies suggest that Ia antigens exist on only a subpopulation of peritoneal macrophages. Alternatively, all cells in the population might bear small amounts of Ia antigens with only a fraction having sufficient numbers of molecules to be detected by the assay systems used.
Studies with B‐Cell Allo‐ and Hetero‐Antisera: Parallel Reactivity and Special Properties
Authors: WINCHESTER R. J. , WANG C. Y. , HALPER J. , HOFFMAN T.
Published: June 2006

The expression of B‐cell antigens on various cell populations was studied through the use of human alloantisera and with heteroantisera raised to preparations of the alloantigen bearing molecules isolated from B‐cell lines. The allo‐ and heteroantisera competed with each other in blocking experiments and gave closely parallel results, reacting with normal and leukemic B lymphocytes, monocytes, K‐rosette‐negative acute lymphatic leukemias, all acute and certain chronic myclogenous leukemias, and a minor population of cells in fetal spleen and liver. These highly immunogenic surface components appeared to comprise the dominant B‐cell‐specific plasma membrane determinants. Neither type of antiserum reacted with any but a minor population of normal or pokeweed‐mitogen‐transformed T cells, fetal thymic lymphocytes, E‐rosette‐positive acute lymphatic leukemias, or Sezary‐cell leukemia. Through the use of these antisera evidence was obtained that Fc‐receptor‐bearing Ig‐negative lymphocytes were divisible into two groups according to the presence or absence of the B‐cell antigens. Both hetero‐ and allo‐antisera blocked binding of immune complexes or antibody‐coated ox erythrocytes to Fc receptors on B cells. F(ab')2, fragments of the hetero‐antibodies strongly inhibited antibody‐dependent cell‐mediated killing.
Antibody‐Dependent Lymphocyte‐Mediated Cytotoxicity
Authors: SIMCHOWITZ L. , SCHUR P. H.
Published: June 2006

The effector cells in antibody‐dependent lymphocyte‐mediated cytotoxicity (ADLMC) have been studied. The cells do not adhere to glass and bear Fc receptors independent of surface Ig and complement receptors. Whereas monocytes have IgG1 and IgG3 Fc receptors, these Fc receptor lymphocytes appear to have receptors for aggregates of all four IgG subclasses.
The Effect of Human Chorionic Gonadotropin and Placental Lactogen on Lymphocyte Transformation In Vitro
Authors: MORSE J. H.
Published: June 2006

Crude preparations of human chorionic gonadotropin (hCG) inhibited phytohemagglutinin (PHA) and allogeneic cell (MLC) lymphocyte stimulation of human peripheral blood lymphocytes (PBL) in vitro as measured by [3H]thymidine incorporation, whereas purified hCG was not inhibitory except at very high concentrations. Sephadex G‐100 chromatography of crude hCG showed that the inhibitory effect on PHA lymphocyte stimulation eluted in those fractions after hormonally active hCG. In addition, purified human placental lactogen (hPL) did not inhibit lymphocyte transformation except at very high concentrations. These data suggest that the immunosuppressive activities found in crude preparations arc not due to the hormones but to contaminating material.
Short‐Term Tissue Culture of Two Nonsecretory Human Myelomas
Authors: NILSSON K. , KILLANDER D. , KILLANDER J. , MELLSTEDT H.
Published: June 2006

Bone marrow cells from two patients without detectable monoclonal immunoglobulin (Ig) in serum and urine but with the clinical picture of plasma cell myeloma were cultivated in vitro. Immunofluorescence studies of cultured living and fixed bone marrow cells showed no signs of Ig production in one of the cases, whereas in the other case cytoplasmic kappa chains were detected, which, however, were not expressed at the surface of living cells. Cells from the latter patient were also subjected to kinetic, ultrastructural, and functional studies in vitro. The fraction of myeloma cells incorporating tritiated thymidine in vitro decreased gradually during prolonged culture, indicating a continuous cell death. The morphological characterization revealed many similarities between this nonsecretory myeloma and classical myelomas, although the frequency of cells with cisternae of endoplasmic reticulum distended by a granular material was unusually high, as was the frequency of ‘flaming’ myeloma cells. ‘Flaming’ cells were not labeled by tritiated thymidine, suggesting that they are nonproliferative end cells. Studies of the Ig synthesis by gel diffusion analyses of supernatant and cell lysates from [14C]leucine‐labeled cultures agreed with the immunofluorescence studies that the myeloma cells were nonsecretory.
Carcinoembryonic Antigen and Cystic Fibrosis Protein in Blood from Cystic Fibrosis Homozygotes and Heterozygote Carriers
Authors: WILSON G. B. , BURDASH N. M. , ARNAUD P. , MONSHER M. T. , FUDENBERG H. H.
Published: June 2006

Carcinoembryonic antigen (CEA) activity was measured by radioimmunoassay in blood from cystic fibrosis (CF) homozygotes, heterozygote carriers of CF, normal healthy controls, and other patient controls with carcinomas involving gastrointestinal organs. All samples were also screened by electrofocusing for cystic fibrosis protein (CFP), a metabolic marker previously shown to be associated with the CF gene. Significantly increased levels of CEA activity were found in all CFP‐positive groups; however, with one exception all patient controls with marked increases in CEA activity were CFP‐negative. Immunodiffusion of perchloric acid extracts of CEA‐like material from heterozygote carrier blood indicated that the CEA‐like material, which was elevated in homozygotes and heterozygotes for CF, showed only partial identity with two separate CEA preparations obtained from colon carcinomas and was not identical to either A, B, or O (H) blood group substances. This glycoprotein material did, however, react with three different anti‐CRA antisera. Our finding of an abnormally increased glycoprotein in cystic fibrosis, taken together with previous reports demonstrating abnormalities in the carbohydrate portion of glycoproteins found in various exocrine secretions in CF, further suggests that the primary defect in this disease is manifested partly as a defect in glycoprotein metabolism. This defect may result from an abnormality in one or more of the glycosyltransferases, possibly caused by a more primary defect in polyamine metabolism.
Characterization of Anti‐Lymphocyte Antibodies in Inflammatory Bowel Disease
Authors: HENDERSON C. A. , GREENLEE L. , WILLIAMS R. C. , STRICKLAND R.G.
Published: June 2006

Sera from patients with inflammatory bowel disease (IBD) showing broad lymphocytotoxic or lymphocyte‐binding activity were subjected to additional analysis to further characterize their properties. Lymphocytotoxins appear to be antibodies predominantly of IgM class as determined by [1] 2‐mercaptoethanol sensitivity, [2] serum fractionation by sucrose density gradient sedimentation, Sephadex G‐200 filtration, and DEAE ion exchange chromatography, and [3] absorption by anti‐Ig immunoadsorbent columns. Lymphocyte‐binding antibody was found to be IgG, IgA, and IgM, as determined by indirect immunofluorescent staining of acetone‐fixed lymphocytes. Individual IBD sera showed marked variability in occurrence of cytotoxic and binding antibodies when tested against the same donor panel of lymphocytes. These studies emphasize the marked heterogeneity of anti‐lymphocyte antibodies occurring in IBD.
Production of Monospecific Antisera Against Antigenic Components ofMycobacterium bovis(BCG)
Authors: HARBOE M. , CLOSS O. , DEVERILL J.
Published: June 2006

Monospecific antisera against four different antigenic components ofMycobacterium bovis(BCG) were produced by immunization of rabbits with precipitates cut out of gels after crossed immunoelectrophoresis (CIE). An important feature is to use a semipurified mycobacterial antigenic preparation, or a cross‐reactive antigen, to obtain a simpler pattern in CIE so that a clean precipitate, free from contaminating antigens, can be obtained for immunization. The findings with one particular antigen are shown, and the use of such antisera is discussed.
Isolation and Characterization of Sea Star Factor
Authors: PRENDERGAST R. A. , LIU S. H.
Published: June 2006

Sea star factor (SSF) harvested from the echinoderm Asterias forbesi celomocytes is a potent inhibitor of the primary immune response to T‐dependent antigens and also suppresses concanavalin‐A‐induced mitogenesis. We have isolated and purified a basic protein from this source which has a molecular weight of approximately 38,000 daltons and is responsible for these biological activities. After complete reduction, SSF was shown to be composed of a single pair of heavy and light chains. Preincubation of spleen cells with concanavalin A does not inhibit binding of SSF; however, specific binding can be inhibited by preincubation with unlabeled SSF. These data suggest that the binding site for SSF is not the same as that for concanavalin A.
Maternal Immunocompetence II. Proliferative Responses of Maternal Lymphocytes In Vitro and Inhibition by Serum from Pregnant Rats
Authors: HARRISON M R
Published: June 2006

Peripheral blood and splenic lymphocytes from interstrain (L × BN) or intra‐strain (L × L) primigravida rats were equivalent to those from virgin L female in their in vitro DNA synthetic responses to paternal strain cells (BN), to unrelated allogeneic cells (ACI), and to the mitogen phytohemagglutinin (PHA). Heat‐inactivated serum from pregnant L rats, when compared to serum virgin or postpartum L rats, regularly suppressed the in vitro response of L lymphocytes to paternal and allogeneic cells. The response of L cells to PHA was not suppressed. The degree of inhibition was related to the final concentration of pregnant serum in culture, concentrations above 2% producing more than 80% inhibition mixed lymphocyte reaction. The inhibiting sera were not cytotoxic by a sensitive 51Cr release assay. Histoincompatibility between mother and fetus is not required for production of this inhibitory effect since it is consistently present in intrastrain (L × L) pregnant rats. Among interstrain pregnant rats, the degree of inhibition is influenced by, but not Specific for paternal‐strain alloantigens. Thus, the proliferative function of thymus‐derived (T) lymphocytes from pregnant intrinsically normal, but the response to allogeneic cells can be altered by factors present in pregnant serum.