bluuebunny/crossref_metadata_2025 huggingface dataset:
Cell Surface Immunoglobulin
Authors: Melcher Ulrich , Uhr Jonathan W.
Published: December 2022

125I-membrane IgM, 125I-membrane IgD-like molecules, and their constituent chains from iodinated murine splenocytes were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The apparent m.w. of the heavy chains decreased as the acrylamide concentration was raised. The membrane µ-chain had a slower mobility than did µ-chain from secreted IgM. Unreduced IgM and IgD-like molecules had mobilities consistent with an H2L2 structure. Intact IgD-like molecules were replaced after overnight dialysis by molecules with the properties of HL. Unreduced surface IgM had a slower mobility than that of monomeric IgM obtained by partial reduction of secreted IgM.
The Class-Specific Immunoglobulin Composition of Fluids Obtained from Various Levels of the Canine Respiratory Tract
Authors: Kaltreider H. Benfer , Chan Maria K. L.
Published: December 2022

Sensitive and specific radioimmunoassays capable of accurately measuring the three major classes of canine immunoglobulins in the 50 to 200 ng/ml range were developed. The concentrations of IgG, IgA, and IgM in samples of fluid obtained from various levels of the canine respiratory tract (stimulated saliva, tracheal washes, and bronchopulmonary lavage supernatants) and in serum were determined by using the radioimmunoassays. The composition of the immunoglobulin classes, expressed as per cent of the total immunoglobulin content of each sample, differed markedly among the respiratory tract-derived fluids and between respiratory fluids and serum. Pilocarpine-stimulated saliva contained predominantly IgA while bronchial washes contained predominantly IgG. The contents of IgA and IgG in tracheal washes were intermediate between those of stimulated saliva and bronchial washes. There appeared to be a progressive decrease in the content of IgA (83% to 19%) and a progressive increase in the content of the IgG (6.3% to 75%) as sampling proceeded from the upper to the lower respiratory tract. The mean IgG/IgA ratios were as follows: stimulated saliva, 0.1; tracheal washes, 2.1; bronchial washes, 4.1; and serum, 23.2. All respiratory tract-derived fluids contained small amounts of IgM. Local and systemic immunization with sheep erythrocytes failed to alter the per cent composition of immunoglobulins in bronchial washes during the primary immune response. The results demonstrate major differences in the composition of immunoglobulins derived from the upper as opposed to the lower respiratory tract. The findings suggest that functional differences may exist in the immunologic apparatus associated with various levels of the respiratory tract.
In Vitro Response of Subpopulations of Human Tonsil Lymphocytes
Authors: Delespesse G. , Duchateau J. , Gausset Ph. , Govaerts A.
Published: December 2022

Human tonsil lymphocytes have been separated into three subpopulations of cells: purified B cells and two subsets of purified T cells (F1 and F2). B cells were obtained by rosetting with neuraminidase treated SRBC. F1 and F2 were separated by filtration on a nylon wool column using different speeds of elution. Purified B cells contained less than 5% T cells, the T cells preparations contained less than 5% B cells for F1 and 10 to 15% for F2, respectively. A significant contamination in cells not identified by any B or T marker was observed in purified B cells and in F1. Adherent cells enhanced the response of each lymphocyte population to PHA and Con A. This explained the paradoxically low responsiveness of the purified T cells. Purified B cells did not respond to these mitogens in different culture conditions. However, a small B cell response was observed when they were cultured in the presence of mitomycin-treated T cells. Striking was the enhancing effect of B cells on the T cell response to PHA and Con A. This enhancing effect was observed even when B cells were treated with mitomycin or depleted in adherent cells. The comparison of the F1 and F2 response suggested that they contained distinct types of T cells.
Neuraminidase Reversal of Resistance to Lysis of Herpes Simplex Virus-Infected Cells by Antibody and Complement
Authors: Tompkins W. A. F. , Seth P. , Gee S. , Rawls W. E.
Published: December 2022

The susceptibility to lysis by antibody and complement was examined in four human cell lines. The cells were infected with herpes simplex virus type 1 and lysis was assessed by the 51Cr release test by using antibodies to herpes simplex virus and guinea pig serum as a source of complement. The four cell lines were found to differ in their susceptibility to lysis, although virus replication was readily demonstrated in the different cell lines. By indirect immunofluorescence, no differences in the expression of virus antigens at the surface of the cells could be found between the different cell lines. Treatment of cells with neuraminidase markedly enhanced the sensitivity of the cells which were relatively insensitive to lysis. The enhancement of susceptibility to lysis by neuraminidase occurred if cells were treated before reaction of the cells with antibody and if the cells were reacted with antibody before treatment with the enzyme. No enhancement was observed when cells were reacted with antibody and complement before neuraminidase treatment. Neuraminidase treatment did not seem to enhance appreciably the quantity of antibody which reacted at the cell surface. The observations suggest that surface properties of certain cells render the cells resistant to lysis by antibody and complement and that the resistance to lysis can be abrogated by treating the cells with neuraminidase.
Activation of C1r by Proteolytic Cleavage
Authors: Ziccardi Robert J. , Cooper Neil R.
Published: December 2022

C1r was unable to cleave and activate proenzyme C1s unless first incubated at 37°C in the absence of calcium before the addition of C1s. The acquisition of ability to activate C1s was associated with, and paralleled by, cleavage of each of the two noncovalently bonded 95,000 dalton chains of the molecule into disulfide linked subunits of 60,000 and 35,000 daltons, respectively. Thus, C1r is converted from an inactive form into an enzyme, C able to cleave and activate C1s by proteolytic cleavage in marked analogy to the activation of several other complement enzymes. Trypsin was also found to cleave C1r but at a different site, and its action did not lead to C1r activation.

C1r activation was inhibited by calcium, polyanethol sulfonate, C1 inactivator, and DFP but not by a battery of other protease inhibitors. C1 inactivator inhibited C by forming a complex with C1r via sites located on the light chain of the molecule. In other studies, cleavage of C1r was not accelerated by the addition of C or C1s.

C1r and C were found to have the same m.w., sedimentation coefficient, and diffusion coefficients. They differed, however, in charge with C1r migrating as a β-globulin and C as a γ globulin on electrophoresis in agarose. The amino acid composition of C1r and of each of the two polypeptide chains of C was determined. Both chains contained carbohydrate.

Proteolytic cleavage of the C1r molecule was found to occur on addition of aggregated IgG to a mixture of C1q, C1r, and C1s in the presence of calcium. Neither C1q, C1s nor aggregated IgG alone, nor C nor C induced C1r cleavage. Liquoid, an inhibitor of C1 activation, inhibited C1r cleavage. Thus, proteolytic cleavage of C1r appears to be a biologically meaningful event occurring during the activation of C1.
The Role of Lymphotoxin in Target Cell Destruction Induced by Mitogen-Activated Human Lymphocytes in Vitro
Authors: Kramer Sandra L. , Granger Gale A.
Published: December 2022

The in vitro destruction of phytohemagglutinin (PHA) coated β L cells by non-immune human lymphocytes was resolved into two distinct phases—lymphocyte dependent and lymphocyte independent. The initial or lymphocyte-dependent phase occurred within the first 2 hr and proceeded equally well at 34 and 37°C. The amount of lymphotoxin (LT) secreted by PHA-activated human lymphocytes in vitro to PHA stimulation was the same at 34 and 37°C. Antiserum and complement inactivation of the aggressor lymphocytes at various intervals revealed that target cell lysis was lymphocyte independent. However, the latter phase was temperature dependent, i.e., proceeding at the permissive temperature of 37°C, but inhibited at the restrictive temperature of 34°C. Further experiments revealed that LT-induced destruction had the same temperature sensitivity as target cell cytolysis occurring during the lymphocyte-independent step. Trypsin treatment of target cells during an early period of the lymphocyte-independent phase protected the target cell from subsequent death, indicating the aggressor lymphocyte has deposited a cytotoxic effector material on its surface. These results suggest the lymphocyte-dependent stage involves the processes required for the induction of LT synthesis and secretion. The actual cytolysis occurring during the lymphocyte-independent stage may be caused by LT or LT-like material(s) deposited on the target cell surface by the mitogen-activated human lymphocyte.
Isolation of Stable and Unstable (“Suppressed”) β2-Microglobulin-Deficient Clones from a Lymphoid Cell Line
Authors: Pious Donald , Soderland Carl , Weagant Stephen , Poulik Dave
Published: December 2022

After selection with anti-β2m and C against a human lymphoid line, about 10% of surviving clones manifested decreased cytotoxic sensitivity to anti-β2m (“suppression”) which gradually reverted to normal after 35 to 55 cell doublings. Antiserum alone induced suppression. Exposure of suppressed clones to DMSO substantially reversed suppression. One clone, surviving selection, had a stable decrease in anti-β2m sensitivity; it possibly represents a β2m mutation.
Immune Serum-Mediated Cytotoxicity Against Trypanosoma Rhodesiense
Authors: Diggs Carter , Flemmings Barbara , Dillon James , Snodgrass Ralph , Campbell Gary , Esser Klaus
Published: December 2022

The metabolic integrity of Trypanosoma rhodesiense can be assessed in vitro by the extent of incorporation of radiolabeled leucine into trichloroacetic acid precipitable material or into material retained after filtration on a glass fiber filter. Incorporation is an approximately linear function of time, and the rate of incorporation is linearly dependent on cell concentration in the presence of normal rat serum. Incorporation is completely prevented if the organisms are reacted with fresh serum from animals immunized with gamma-irradiated parasites; the degree of inhibition is a function of the dose of immune serum used. This serum-mediated cytotoxic activity is abrogated by heating the serum, but can be fully restored by addition of fresh rat or guinea pig serum to the heated immune serum. The serum activity arises promptly after one to four immunizing doses of irradiated parasites, falls to lower levels by 1 month, but persists for at least 2 ½ months, and is unaffected by challenge with viable trypanosomes.
Synthesis of Gm Allotypes by Human Tonsillar Lymphocytes in Culture: Concordance with Serum Phenotype
Authors: Litwin Stephen D. , Hütteroth Thomas H. , Balaban Linda
Published: December 2022

Isolated human tonsillar lymphocytes were cultured with pokeweed mitogen, phytohemagglutinin, and without mitogen for 9 to 28 days. IgK, Gm(a) and Gm(f) were then quantitated in the cell suspensions. In cultures of cells derived from persons whose blood was heterozygous for IgGl allotype antigens Gm(a+f+), approximately equal amounts of Gm(a) and Gm(f) were found. In cultures of cells of Gma or Gmf homozygotes, there was complete concordance between the Gm allotype antigens produced by the cultures and the donor's serum phenotype—with no instance, either at zero time or at culture termination in which a Gm antigen was detected which was absent from the donor's serum. It was concluded that in vitro genetic allotype synthesis in tonsillar lymphocytes during short-term culture mirrored accurately in vivo Gm expression.

IgK and Gm antigen synthesis was highest in the flasks containing pokeweed mitogen although both phytohemagglutinin and no-mitogen control flasks showed, in certain experiments, proliferation and an increase in the Ig per viable cell. It was observed that no-mitogen flasks contained twice as much allotype antigen as did phytohemagglutinin flasks suggesting an inhibition of Ig synthesis associated with the mitogen. The tonsillar lymphocytes, under the experimental conditions employed, were shown by a radio-incorporation and immunoprecipitation technique to be synthesizing polyclonal Ig de novo, at the termination of the cultures.
The Defect in µ Heavy Chain Disease Protein GLI
Authors: Franklin Edward C. , Frangione Blas , Prelli Frances
Published: December 2022

Amino acid sequence studies of µHCD protein GLI reveal the absence of the amino terminal 130 residues. The molecule starts at Ala 131 and contains Cys 140, normally involved in the H-L disulfide bridges. Except for the deletion and presumably an extra H-H disulfide bridge involving Cys 140, the remainder of the µ-chain appears normal.
Factors Influencing the Response of Mice to Type III Pneumococcal Polysaccharide
Authors: Warr Gregory W.
Published: December 2022

Sir: I would like to comment on a number of issues raised by your correspondents Baker and Prescott (1), in their letter entitled “The Basis for Conflicting Results Obtained in Studies on the Plaque-Forming Cell Response to Type III Pneumococcal Polysaccharide.”

Their argument can, I believe, be summarized in the following points.

Anti-lymphocyte serum (ALS)-induced enhancement of the response to Type III pneumococcal polysaccharide (SIII) has only been observed (2–4) by using indicator erythrocytes sensitized, by the chromic chloride method, with purified SIII (5), but not by using indicator erythrocytes sensitized with the filtrate from a culture of pneumococci (6–8) as described originally by Askonas et al. (9). This culture-filtrate method of sensitizing target erythrocytes detects primarily response to a contaminating antigen present in small amounts in the immunizing preparation of SIII, but present in large amounts in the culture filtrate.
Antibody-Dependent Cell-Mediated Cytotoxicity in Humans
Authors: Trinchieri Giorgio , De Marchi Mario
Published: December 2022

Different classes of protease inhibitors and substrates were tested for their effect on the ability of human lymphocytes to mediate antibody-dependent cytotoxicity (Ab CMC). All the inhibitors tested (serine esterase inhibitors, chloromethyl ketone derivatives of tosyl-amino acids, synthetic protease substrates), except for the naturally occurring protease inhibitors (derived from soybean, lima bean, and porcine pancreas), were able to suppress, or to reduce significantly, the cytotoxicity. In the absence of a direct demonstration of an esterase activity, sensitive to the action of the inhibitors in the effector lymphocytes, careful controls were used to restrict the possibility that some nonspecific effect of the drugs was being interpreted. Particularly, the dependence of the inhibition of cytotoxicity as an effect of drugs on membrane transport mechanisms or on energy metabolism was excluded. The similarity between results obtained with compounds of different chemical characteristics and different molecular mechanisms of action supports a specific effect of the inhibitor on cellular esterase(s) or possibly protease(s). The fully reversible inhibition obtained with serine esterase inhibitors suggests that the relevant enzymes are activated only after effector-target cell interaction; the irreversible effect of chloromethyl ketone derivatives, however, does not allow the participation of already activated enzymes to be excluded.

The results presented in this study on the probable role of cellular esterases, on cation requirement and on the sequence of biochemical steps in Ab CMC add a new element to the analogy between this cellular phenomenon and different types of cytotoxicity or other immunologically induced cellular reactions, suggesting that the biochemical mechanisms of cytotoxicity may partly reflect a common pattern of cellular response to external stimuli.
Characterization of T and B Antigen-Binding Cells for β-Galactosidase
Authors: Swain Susan , Modabber Farrokh , Coons Albert H.
Published: December 2022

The results reported in this paper demonstrate that the enumeration of cells binding β-galactosidase (Z) as an antigen, revealed by subsequent substrate hydrolysis, is an excellent method for the detection and study of antigen-binding cells (ZBC). The binding found is specific and is restricted to a small number of lymphocytes that bind a large number of Z molecules via surface receptors. Such ZBC were found at mean frequencies of 150 per 106 in the thymus and 200 to 300 per 106 in the spleen. The binding cells of both organs were heterogenous with individual ZBC binding from 105 to 106 molecules of enzyme as determined by substrate hydrolysis, although this might well be an overestimate of the number of actual receptors. The profiles for the frequency of ZBC binding different numbers of molecules were nearly identical for thymus and spleen, in contrast to descriptions of the binding of many other antigens. Receptors responsible for Z binding appear to be superficially located on the cell since they are trypsin-sensitive to a large extent and are not increased by fixation.
Age-Associated Involution of Cellular Immune Function
Authors: Pachciarz Judith A. , Teague Perry O.
Published: December 2022

Autoimmune-susceptible (A/J and C57BL/6J) and non-autoimmune-susceptible (DBA/1J) strains of mice were adult thymectomized (aTx). Groups of mice, treated by aTx, aTx and 330 rads (R), aTx and 650 R, and 330 R or 650 R only, were compared to normal age-matched control (normal and sham Tx) mice. T cell functions were then followed as measured by in vitro spleen cell responsiveness to the T cell mitogens phytohemagglutinin (PHA) and staphylococcal enterotoxin B (SEB) in a long-term study. A decline in spleen cell responsiveness with natural aging up to 18 months of age was seen to PHA in all three strains and to SEB in A/J and C57BL/6J mice. Adult Tx alone was followed by an accelerated decline in spleen cell responsiveness to PHA and SEB in all three strains of mice. Irradiation combined with aTx-reduced spleen cell responsiveness to SEB in A/J and DBA/1J mice. Irradiation with or without aTx diminished the bone marrow stem cell pool as measured by exogenous colony-forming units, and the hematocrit of all three strains of mice. Thus, the natural decline in cellular immunity with aging was accelerated by aTx in both autoimmune-susceptible and non-autoimmune-susceptible strains of mice. These results emphasize that physiologic thymic function(s) must continue throughout life in order to maintain T cell function.
IgM Complex Receptors on Subpopulations of Murine Lymphocytes
Authors: Lamon E. W. , Andersson B. , Whitten H. D. , Hurst M. M. , Ghanta V.
Published: December 2022

Murine lymphocytes from spleen, lymph node, and thymus were examined for IgM complex receptors. Lymphocytes from all three organs were found to bind SRBC sensitized with IgM from various sources including: primary anti-SRBC serum, murine and rabbit anti-Escherichia coli LPS sera, and a murine IgM myeloma (MOPC 104E). Rosette formation by lymphocytes with IgM-sensitized SRBC was inhibited by soluble antigen-IgM complexes but not by IgM or antigen alone. Rosette formation was also inhibited by human IgM (Fc)5µ but not by Fab µ. Antiserum and complement treatment of the cells and subsequent recovery of the viable cells by trypsinization, filtration, and washing revealed the IgM rosette-forming cell (RFC) in the thymus to be a T cell. Spleen on the other hand was found to contain both B and T cells capable of binding IgM sensitized SRBC. Removal of both B and T cells from spleen cell suspensions eliminated all IgM RFC. The IgM complex receptor was found to be trypsin insensitive. Anti-Ig column fractionation enriched IgM RFC in spleen and lymph node suspensions passed through the columns, whereas cells bearing surface Ig, IgG complex receptors, and C3 receptors were retained in the columns.
Purification and Characterization of a Liver-Specific Antigen
Authors: Mihas Anastasios A. , Hirschowitz Basil I. , Saccomani Gaetano
Published: December 2022

A liver-specific antigen (F-antigen) previously demonstrated in saline extracts of BALB/c mouse liver by double immunodiffusion was isolated and characterized. The antigen was found widely distributed among mammals but absent from avian and frog liver extracts. In immunoelectrophoresis it had an electrophoretic mobility similar to that of serum β2-globulins, was relatively thermolabile, and was precipitated at 30 to 70% saturated ammonium sulfate concentration. Evidence was presented that this antigen is a protein or a moiety closely associated with protein. Gel-filtration on Sephadex G-200 revealed liver-specific antigenicity in the second peak. Ion-exchange chromatography on DEAE-Sephadex A-50 revealed four peaks of which only the third one exhibited liver-specific antigenicity. This active peak contained 11 polypeptides on SDS polyacrylamide gel electrophoresis. After electrophoresis on acrylamide gel in the absence of SDS, antigenic activity was detected on one fast-moving band. Extraction of the protein band followed by SDS gel electrophoresis showed one major component of m.w. 75,000 and two minor bands of m.w. 72,000 and 93,000, respectively.
Reaginic Antibody Formation in the Mouse
Authors: Takatsu Kiyoshi , Ishizaka Kimishige
Published: December 2022

Intravenous injections of urea-denatured ovalbumin (UD-OA) into OA-primed high responder mice suppressed the antibody response not only to the priming antigen but also to subsequent immunization with dinitrophenyl derivatives of OA (DNP-OA). The transfer of normal spleen cells or OA-primed spleen cells into UD-OA-treated animals did not restore the capacity of responding to DNP-OA to form anti-DNP IgE and IgG antibodies. The transfer of splenic T cell fraction from the UD-OA-treated animals into normal syngeneic mice diminished both IgE and IgG antibody responses of the recipients to DNP-OA. The B cell-rich fraction from the same donors failed to affect the anti-hapten antibody response and enhanced anti-carrier (OA) IgG antibody response of the recipients. It was also found that the transfer of T cell-rich fraction of OA-primed spleen cells failed to suppress antibody response of the recipients to DNP-OA. The results indicated that spleen cells of UD-OA-treated mice contained suppressor T cells which are distinct from helper cells. Suppressive activity of T cells in the UD-OA treated animals was specific for OA. The transfer of the T cell-rich fraction failed to suppress anti-DNP antibody response of the recipients to DNP-KLH.
Changes in C3 Metabolism during Protozoan Infection (Babesia Rodhaini) in Rats
Authors: Chapman William E. , Ward Peter A.
Published: December 2022

Metabolism of the third component of complement (C3) and IgG was measured in rats before and during infection with the hemosporidium agent Babesia rodhaina. In the course of infection, hypocomplementemia and immune complex nephritis developed. During babesial infection, in most animals the half-life of C3 fell sharply, as did serum levels of C3; the catabolic rate for C3 sharply increased, whereas the synthetic rate sharply decreased. In contrast, the catabolic rate for IgG remained unchanged. The alteration in the metabolism of C3 in the face of nonparallel changes in IgG metabolism suggests that abnormal glomerular filtration and increased vasopermeability cannot explain the findings. Babesial infection in the rat provides a useful model for the study of acquired C3 metabolic defects that have been observed in humans with immune complex diseases.
Effects of C-Reactive Protein on Platelet Function
Authors: Fiedel Barry A. , Gewurz Henry
Published: December 2022

C-reactive protein (CRP) is an acute phase reactant which shares numerous functional characteristics with the immunoglobulins. In the present study CRP was found to inhibit the aggregation of human platelets stimulated by either modified human immunoglobulin or thrombin. This effect did not involve chelation of calcium or cytotoxicity, and was overcome by larger amounts of the aggregating agents. CRP also inhibited the activation but not the activity of platelet factor 3 and the release of β-glucuronidase. Thus, CRP can inhibit multiple platelet reactivities. We suggest that this property of CRP may play an important role in the control of platelet responsiveness during reactions of inflammation, defense, and repair.
The Binding of Complement by Complexes Formed Between a Rabbit Antibody and Oligosaccharides of Increasing Size
Authors: Jaton Jean-Claude , Huser Hans , Riesen Walter F. , Schlessinger Joseph , Givol David
Published: December 2022

Immune complexes formed between a homogeneous rabbit antibody to type III pneumococcal polysaccharide and a series of oligosaccharides of varying size derived from it were prepared and tested for their ability to fix guinea pig hemolytic complement.

Antibody and either tetra-, hexa-, or octasaccharide formed only monomeric antibody-hapten complexes and did not show any complement binding. A dodecasaccharide and a 16-sugar residues oligomer formed dimer and trimer immune complexes. These complexes were also unable to fix complement. However, as the size of the sugar oligomers was increased to about 21 sugar residues per oligosaccharide molecule or more, the resulting complexes exhibited substantial complement binding, concomitant with the formation of antigen-antibody aggregates higher than trimers.

On the other hand, an independent study carried out with the same material suggested changes in the conformation of the Fc moiety in the antibody molecule upon addition of oligosaccharide ligands as small as a 16-residue unit. Since the resulting complexes hardly exhibited any complement binding, ligand-induced conformational changes in the Fc part of the antibody molecule appears to be an insufficient condition per se for triggering complement fixation.
The Significance of Varying SRBC/Lymphocyte Ratio in T Cell Rosette Formation
Authors: Chisholm Rex L. , Tubergen David G.
Published: December 2022

The incubation ratio of sheep red blood cells (SRBC) to lymphocytes is a critical factor in rosette formation, whereas the length of time SRBC and lymphocytes are incubated together does not significantly affect the percentage of lymphocytes forming rosettes. The graph obtained by plotting percentage of rosette formation against the ratio of SRBC to lymphocytes is similar to that resulting from the formation of bimolecular complexes. If rosette formation is analogous to formation of bimolecular complexes, maximal rosette formation occurs when the system is saturated, i.e., with excess SRBC, and is a measure of the total capacity of a lymphocyte population to form rosettes. In addition, the percentage of rosette formation observed at a limiting SRBC/lymphocyte ratio gives an indication of the avidity of the lymphocytes for SRBC. This interpretation may provide an explanation for the difference between the “active” and “total” rosettes. When the log of the SRBC/lymphocyte ratio is plotted against percentage of rosette formation, a straight line is obtained, suggesting that within a given normal lymphocyte sample, T cell subsets with different avidities are not detected by rosette formation at different SRBC/lymphocyte ratios.
Macrophage Requirements of CR- and CR+ B Lymphocytes for Antibody Production in Vitro
Authors: Hoffmann Michael K. , Hämmerling Ulrich , Simon Markus , Oettgen Herbert F.
Published: December 2022

A Sephadex G-10 column coated with antigen-antibody complexes and complement retains complement receptor-bearing (CR+) mouse spleen cells. The effluent is rich in thymus-derived cells (T cells), and contains bone marrow-derived cells (B cells) which carry surface immunoglobulin (Ig), Ir-associated antigen (Ia), and Fc receptors, but no complement receptors (CR-). Although both unfractionated and CR- B cell populations are capable of producing antibody to red cell antigens, they differ in their requirements for the initiation of the response. Unfractionated B cells cooperate with primed as well as unprimed helper T cells; macrophages are required for this cooperation but can be replaced by 2-mercaptoethanol. CR- B cells cooperate with primed but not with unprimed T cells provided macrophages are added to cultures. After addition of culture supernatant from BCG-activated macrophages CR- B cells cooperate with both unprimed and primed T helper cells.
Immunologic Mechanisms in the Pathogenesis of Virus-Induced Murine Leukemia
Authors: Proffitt Max R. , Hirsch Martin S. , Ellis David A. , Gheridian Beatriz , Black Paul H.
Published: December 2022

Thymocytes from preleukemic mice persistently infected with Moloney murine leukemia virus (MuLV-M-carriers) were vigorously autoaggressive toward normal syngeneic target cells; they exhibited a graded response to allogeneic cells, but they spared xenogeneic cells or syngeneic cells infected with MuLV-M or MuLV-G (Gross). Syngeneic target cells infected with nononcogenic lymphocytic choriomeningitis virus (LCMV), or transformed by the chemical carcinogen 3-methylcholanthrene were not similarly spared. This phenomenon, apparently induced by MuLV-M, is not associated with all persistent virus carrier states. Thymocytes from mice persistently infected with LCMV or with the lactic dehydrogenase virus (LDHV) failed to demonstrate an autoaggressive behavior. That transplantable lymphoma cells (derived from MuLV-M-carriers) were autoreactive in a pattern similar to thymocytes from preleukemic mice suggests a unique role for MuLV in the events leading from altered recognition of “self” to lymphoma.
Identification of T and B Cell Subpopulations in Human Peripheral Blood: Electrophoretic Mobility Distributions Associated with Surface Marker Characteristics
Authors: Kaplan Joel H. , Uzgiris E. E.
Published: December 2022

The electrophoretic mobility distributions of human peripheral blood lymphocytes isolated on Ficoll-Hypaque gradients were characterized by laser Doppler spectroscopy. Three major subpopulations were spectrally resolved, due to differences in their mobility, when an electric field was applied to the scattering cuvette. The fastest component (centered at 2.35 µm/sec/V/cm for 25°C, 0.28 M sucrose medium of 0.005 ionic strength) passed through nylon fiber columns and was identified as a T cell subpopulation. The slowest component (1.85 µm/sec/V/cm) which was further enriched by a one-step rosette procedure with sheep erythrocytes, reacted with activated-complement (C3) and antiserum to human immunoglobulins and was therefore identified as a B cell subpopulation. The intermediate component (centered at 2.15 µm/sec/V/cm) appears to be another T cell subpopulation. Although cells of this component did not pass through nylon fiber columns, they did rosette with sheep erythrocytes. Furthermore, these cells did not appear to have surface immunoglobulins or complement receptors.
Synchronous Regulation of VH and CH Allotypes among Ig Molecules in Multi-Heterozygous Rabbits Suppressed with Anti-VH Allotype Antibody: Emergence of IgA from Suppression Prior to IgG and IgM
Authors: Eskinazi Daniel P. , Gilman-Sachs Alice , Knight Katherine L. , Dray Sheldon
Published: December 2022

Heterozygous rabbits of genotype a1n81f73g74/a2n82f71g75 were suppressed at birth for the VH region al allotype. At 8 weeks of age, quantitative analysis of serum IgG, IgM, and IgA molecules showed that the VHal specificity was effectively suppressed in the three classes of Ig and that the suppression was extended to the CH region n81 specificity on µ-chains as well as to the CH region f73 and g74 specificities on αf and αg chains. At 26 weeks of age, analysis of serum IgG and IgM molecules showed that a1 was still suppressed to approximately the same extent in both Ig classes and the suppression was still extended to the CH region n81 specificity. However, at 26 weeks, the percentage of molecules with a1 specificity had doubled among serum and colostral IgA molecules and this increase was extended to the CH region f73 and g74 specificities. Thus, the suppressed allotypes reappeared first among IgA molecules. Our data are consistent with a regulatory mechanism which controls and synchronizes the expression of the VHa and the CH allotypes expressed on the same heavy chain. The order of the re-expression of the suppressed allotypes with respect to Ig class may allow further definition of selective regulatory mechanisms for the synthesis of Ig classes.
An Improved Method for Isolation of H-2 and Ia Alloantigens with Immunoprecipitation Induced by Protein A-Bearing Staphylococci
Authors: Cullen Susan E. , Schwartz Benjamin D.
Published: December 2022

Mouse alloantigens, including the histocompatibility (H-2) and immune response linked (Ia) antigen molecules, can be readily isolated by a new precipitation technique utilizing protein A-bearing, fixed Staphylococci. The antigens are prepared by radiolabeling mouse spleen cells with 3H-leucine, solubilizing with the non-ionic detergent Non-Idet P-40, and purifying by affinity chromatography with lentil lectin coupled to Sapharose. The antigen preparations are mixed with appropriate alloantisera, and immune complexes formed in the mixture are then bound by the protein A sites on the Staphylococci. The organisms, Staphylococcus aureus, Cowan I strain (ATCC 12598), are heat inactivated and fixed, and are a stable, uniform IgG-binding reagent, especially when stored frozen. The antigen-antibody complexes are easily eluted from the organisms for electrophoretic analysis. The precipitation mediated by the organisms is more efficient, rapid, and artifact-free than the traditional “sandwich” precipitation technique involving anti-globulin reagents.
Studies on the Immune Response to Fixed Antigens. Preferential Induction of Helper Function with Heavily Trinitrophenylated Sheep Erythrocytes, and Glutaraldehyde-Treated Sheep Erythrocytes
Authors: Kahan Melvyn , Berman-Goldman Ruth , Saltoun Rita , Naor David
Published: December 2022

Mice primed with heavily trinitrophenylated sheep red cells (TNP128SRC) or glutaraldehyde-treated sheep red cells (G-SRC) developed an early helper function mediated by thymus-derived cells. Such mice were able to produce high secondary responses to both hapten and carrier after challenge 2 days after priming, with lightly trinitrophenylated SRC (TNP 0.14SRC). However, the primary response of the TNP 128SRC or G-SRC-primed mice were very low to undetectable, and their secondary responses were also low when the challenge antigen was administered 4 days after priming or later. Inhibitory humoral factor(s) which were induced in the primed animals appeared responsible for the decreased capacity of primed mice to mount a secondary response when challenged later than 2 days after priming. Transfer of spleen cells from TNP 128SRC-primed mice to sublethally irradiated recipients circumvents their exposure to inhibitory humoral factor(s) present in intact animals allowing them to react with challenge antigen. Enriched populations of T cells, but not B cells, were able to transfer this early immunologic memory to irradiated recipients. The theoretical and practical implications of these results are discussed.
Antigenic Analysis of the Major Structural Protein of the Mason-Pfizer Monkey Virus
Authors: Schochetman G. , Boehm-Truitt M. , Schlom J.
Published: December 2022

The major internal protein, p27 (m.w. 27,000 daltons) of the Mason-Pfizer monkey virus (MPMV) was purified by gel filtration and ion-exchange chromatography and then used to develop a radioimmunoassay (RIA). This RIA was specific for MPMV because no immunologic cross-reactivity was observed between p27 of MPMV and 13 different RNA tumor viruses of mammalian and avian origin. However, the p27 of MPMV grown in three different primate cells exhibited identical antigenic cross-reactivity. In addition, significant levels of p27 were found only in MPMV-infected cells. These results indicate that synthesis of p27 is induced after virus infection and that p27 represents a viral-coded protein.
Fine Structure of Three Different Anti-Fluorescein Combining Sites: Induced Circular Dichroism of Hapten Bound to Autologous and Heterologous Recombinants
Authors: Gollogly John R. , Cathou Renata E.
Published: December 2022

We have previously reported the sequential appearance in hyperimmunized rabbits of three distinct types of antifluorescein-combining sites that can be distinguished by the characteristic induced circular dichroism (CD) of the bound hapten, fluorescein. Such induced CD depends on the configuration of the surrounding residues, and, in the case of anti-fluorescein, can be localized to the configuration of the sub-site which binds the hydroxyxanthenone moiety of fluorescein. In the present investigation, we have studied the fine structure of both autologous and heterologous recombinant sites and of the free chains by measuring the induced CD of bound hapten. Heavy chain dimers at pH 5.4 bound fluorescein that displayed a weak negative CD band which is different from that observed in any of the native antibodies. No induced CD was observed with light chains. Thus, the hydroxyxanthenone subsite does not exist intact in any of the isolated chains. Fluorescein bound to purified autologous recombinants, when studied at pH 7.5, exhibited CD spectra very similar to those observed for the original antibodies. Most significantly, fluorescein bound to purified heterologous recombinants, prepared in all possible combinations, showed CD spectra most similar to those exhibited by sites from which the heavy chain was derived. Thus, the microenvironment of the subsite appears to be due primarily to the heavy chain.
Humoral Immunity in Experimental Syphilis
Authors: Bishop Nancy H. , Miller James N.
Published: December 2022

Evidence of a role for humoral factor(s) in immunity to experimental syphilis has been provided by the demonstration that passive immunization of rabbits by daily i.v. injections of immune serum significantly delays the appearance and markedly diminishes the severity and duration of lesions which develop after challenge with Treponema pallidum. Five rabbits were injected daily over 37 days with 3 ml/kg body weight of pooled immune rabbit serum injection with 1.1 × 103T. pallidum, Nichols strain, at each of four sites. The animals developed atypical lesions of short duration after an average delay in onset of 28 days short duration after an average delay in onset of 28 days beyond the development of typical lesions in control animals similarly injected with nonimmune serum or saline. The failure of passive immunization to provide complete protection was evident not only in the development of the atypical lesions, but also in the demonstration of disseminated infection in the tissues of three of the four surviving animals 7 months after challenge. The possibility that incomplete protection may have been due to 1) insufficient immune serum levels, 2) intracellular location of T. pallidum, and/or 3) cell-mediated mechanisms is discussed.
Analysis of Complement-Dependent Antibody-Mediated Lysis of Target Cells Acutely Infected with Measles
Authors: Hicks John T. , Klutch Michael J. , Albrecht Paul , Frank Michael M.
Published: December 2022

A sensitive assay for the detection of complement-dependent cytolytic measles-antibodies in monkey and human sera is described. The high sensitivity of the technique is dependent upon the use of Vero or MA-104 cells acutely infected with a high-titered, non-syncitiogenic strain of attenuated measles virus. Cytolytic antibody titers of serum can be determined either by endpoint dilution in the presence of a standard dilution of heterologous (guinea pig) complement or by percentage of lysis of a specific number of target cells at a standard dilution of immune serum (1:32). The latter technique proved expedient in the comparison of cytolytic activities of large numbers of serum samples. Clearly, the alternative complement pathway is effective in lysis of these cells, but neither the classical nor the alternative pathway alone appears to be as effective as both pathways combined. No prozone effect occurred with homologous complement, and inhibition of lysis was eliminated by washing sensitized cells before addition of heterologous complement. Lysis of measles-infected cells untreated with measles antibody occurred in the presence of both normal and C4-deficient guinea pig serum at high concentrations, but no such activity occurred with non-immune human or rhesus serum. Antibody-mediated complement-dependent lysis of measles-infected cells may be an important immune defense against acute measles infection.
Antibody-Mediated Basophil Accumulations in Cutaneous Hypersensitivity Reactions of Guinea Pigs
Authors: Askenase Philip W. , Haynes J. David , Hayden Betty J.
Published: December 2022

Cutaneous basophil hypersensitivity (CBH) was studied in guinea pigs by using simplified histologic techniques. Animals immunized with oxazolone or picryl conjugates of keyhole limpet hemocyanin (KLH) emulsified with complete (CFA) or incomplete Freund's adjuvant (IFA) were found to have hapten-specific cutaneous basophil reactions when skin tested at 1 week with oxazolone or picryl chloride contact painting or intradermal injection of oxazolone or picryl-conjugated human serum albumin, respectively. Thus, hapten-specific cutaneous basophil reactions were present in guinea pigs immunized with CFA for classical delayed hypersensitivity, and in animals immunized with IFA for Jones-Mote reactions.

Hapten-specific 24-hr cutaneous basophil reactions were passively transferred with immune serum from donors sensitized with conjugates of oxazolone or picryl-KLH in CFA or IFA, and with serum from oxazolone contact-sensitized animals as well. As little as 0.5 ml sera obtained from donors 1 week after immunization could systemically transfer cutaneous basophil reactions. It is likely that hapten-specific cutaneous basophil reactions are mediated by small quantities of serum antibodies.

We conclude that antibody-mediated cutaneous basophil reactions may be distinctive hypersensitivity responses that can be distinguished from classical anaphylactic, Arthus, and delayed hypersensitivities. It is suggested that CBH reactions are heterogeneous and that antibody products of B lymphocytes, and factors probably derived from T lymphocytes, play a role in basophil accumulations at cutaneous hypersensitivity reactions.
Chemotactic Responses of Normal Human Basophils to C5a and to Lymphocyte-Derived Chemotactic Factor
Authors: Lett-Brown Michael A. , Boetcher David A. , Leonard Edward J.
Published: December 2022

Peripheral blood basophils from normal human subjects responded chemotactically to the complement component C5a and also to the supernatant fluid from cultured human lymphocytes stimulated by mitogens or specific antigen. Basophil chemotactic factor (BCF) was detected in stimulated lymphocyte cultures within 24 hr and increased for the first 3 days. BCF was found after stimulation of lymphocytes by B or T cell mitogens as well as by specific antigen. The amount of BCF in culture fluids was independent of cell proliferation. Monocyte chemotactic factor (MCF) and BCF could not be separated from one another by Sephadex or DEAE chromatography. Both activities were eluted from Sephadex G-100 columns in a m. w. region of about 15,000 daltons and were found after ion exchange chromatography on DEAE in a peak eluted by 0.1 M NaCl. These findings suggest that mitogen or antigen-stimulated lymphocytes secrete a molecule that is chemotactic for both monocytes and basophils. Thus, in the absence of a basophil-specific chemotactic molecule, the preferential accumulation of basophils in certain types of delayed hypersensitivity skin reactions must be due to additional factors such as basophil chemotaxis augmentation factor.
The Xenogeneic Effect
Authors: Farrar John J. , Fuller-Bonar Janet
Published: December 2022

Studies were conducted on the mechanism of action of a soluble mediator which was generated in human mixed lymphocyte cultures and assayed for helper activity in T cell-deficient murine spleen cell cultures. The mediator, termed xenogeneic reconstitution factor or XRF, restored the anti-sheep erythrocyte plaque-forming cell response of spleen cells from thymectomized, lethally irradiated, and syngeneic bone marrow transplanted (TxB) mice. It was found that in order to obtain a maximal antibody response, the XRF had to be present sometime during the first 24 to 40 hr of the induction process. XRF was also required during the last 24 hr of the incubation period. A striking synergistic effect was obtained by combined exposure of the T cell-deficient spleen cell cultures to XRF during the first and last days of culture. These data suggested a bi-modal mechanism of action of XRF, and raised the possibility that in addition to providing a signal to the B cells, XRF-mediated activation of the residual TxB splenic T cells was crucial to the successful restoration of the antibody response. Treatment of the nonadherent splenocytes from the TxB mice with anti-T cell serum and guinea pig complement completely abrogated the antibody response of these cells in the presence of adherent spleen cells, sheep erythrocytes, and the helper factor XRF. The antibody response of this combination of cells, antigen, and XRF was reconstituted by a population of unimmunized murine T cells which, in the absence of XRF, were totally unable to restore responsiveness. These results suggest that B cell activation to the formation of antibody involves the mandatory co-participation of two functionally distinct helper activities.
Enhancement of Early Human E Rosette Formation by Cholinergic Stimuli
Authors: Galant Stanley P. , Lundak Robert L. , Eaton Linda
Published: December 2022

The effects of the cholinergic stimuli carbamylcholine (carbachol) and dibutyrl cyclic guanosine monophosphate (DBCGMP) were determined on both ‘early’ and ‘total’ E rosette formation. Ficoll-Hypaque-separated lymphocytes were preincubated with either carbachol or DBCGMP over a 10-3 M to 10-13 M dose range. Both agents significantly enhanced ‘early’, but not ‘total’ E rosette formation. Peak enhancement above control values occurred at 10-7 M (72%) and 10-9 M (69%) for carbachol and 10-5 M (70%) and 10-7 M (70%) for DBCGMP. Kinetic studies showed a rapid onset of enhancement (2.5 min) for carbachol, whereas DBCGMP required 15 min for significant enhancement to occur. The muscurinic nature of carbachol enhancement of E rosettes was demonstrated. Atropine at 10-7 M completely abolished the carbachol effect while showing little inhibition of the DBCGMP effect on rosette formation.

These studies indicate that the cholinergic stimuli carbachl and DBCGMP significantly enhance the ‘early’ E rosette former in man. Human T lymphocytes appear to have functional cholinergic receptors that can be blocked by the muscurinic antagonist atropine. The role of the cyclic nucleotides and their stimulants on the immune system is incompletely understood, but it would appear that they are extremely important in the differentiation and function of the T lymphocyte. E rosette formation may be a useful model in man for studying the effects of the cyclic nucleotides on the human T lymphocyte.