bluuebunny/crossref_metadata_2025 huggingface dataset:
DRw Antisera React with Activated T Cells
Authors: DeWolf William C. , Schlossman Stuart F. , Yunis Edmond J.
Published: December 2022

Nylon wool-purified T cells appear to be nonreactive in a lymphocytotoxicity assay with HLA-DRw antisera and complement before cell activation. However, after activation in mixed lymphocyte culture, responder cells express determinants that are strongly reactive with DRw alloantisera after 6 days and gradually disappear by 16 to 18 days. Restimulation of the primed cells resulted in re-expression of the blast determinants. Mitogenic stimulation with Con A or purified PHA (HA-17) also resulted in temporary expression of these determinants; reactivity usually conformed to DRw genetic restriction; however, occasional extra reactions occurred that were variable depending on the method of activation (i.e., MLC, Con A, or HA-17). The results suggest the presence of additional allospecificities within some of the DRw antisera that react with “Ia-like” antigens on activated cells from unique subsets of T cells. Whether these DRw antisera contain antibodies against T cells or agains activation or differentiation T cell antigens is not as yet clear.
Human B Cell Differentiation
Authors: Bich-Thuy Le Thi , Brochier Jean
Published: December 2022

In vitro human plasma cell generation induced by both T-dependent (PWM) and T-independent (NWSM) mitogens was found to be suppressed by peripheral blood lymphocytes preincubated with human aggregated IgG. T cells, but not B lymphocytes, were able to mediate the suppressive activity; since aggregated (Fab)′2 fragments were found unable to generate suppressor cells, it was concluded that the suppressor cell was a T lymphocyte bearing Fcγ receptors. These cells appeared to be largely radiosensitive. In most cases the proliferative responses remained unchanged. Since NWSM-induced activation is not dependent on the presence of T cells, these results show that, at least in this case, T cells exert their suppressor function directly on B lymphocytes. Whether PWM-induced B cell differentiation is suppressed by the same mechanism or/and by inactivation of T helper lymphocytes remains under investigation.
Somatic Cell Hybrids of Mouse Myeloma Cells and B Lymphocytes from a Patient with Agammaglobulinemia: Failure to Secrete Human Immunoglobulin
Authors: Schwaber Jerrold F. , Rosen Fred S.
Published: December 2022

Somatic cell hybrid clones were isolated from the fusion of RPC5.4 mouse myeloma cells and B lymphocytes from a patient with common varied agammaglobuinemia. The patient has B lymphocytes that synthesize immunoglobulin but fail to secrete immunoglobulin. The hybrid character of the six clones was established by examination of metaphase chromosome spreads. Most of the hybrid clones expressed mouse and human surface immunoglobulin. All of the clones synthesized immunoglobulin of mouse and human parental origin. Mouse parental immunoglobulin was secreted, whereas the human parental immunoglobulin was not secreted. Human light chain molecules were secreted as part of hybrid H2L2 molecules formed with mouse heavy chains. Human heavy chains had a reduced m.w. in comparison to the mouse heavy chains. Kinetic experiments indicated that human Ig was synthesized in amounts that were comparable to the mouse Ig. Pulse-chase experiments showed that that the intracellular human Ig was removed from the cytoplasm, probably by degradation. These experiments demonstrate that the hybrid cells are an in vitro model of naturally occurring failure of immunoglobulin secretion from agammaglobulinemia. The failure of fusion with mouse myeloma cells to complement the secretion defect suggests that these B cells produce an altered immunoglobulin molecule that is not programmed for secretion.
Mechanisms of Leukemogenesis
Authors: Schenk Paul J. , Howe Michael L.
Published: December 2022

Examined in this paper is the capacity of 334C murine leukemia virus (MuLV) to stimulate the generation of virus-specific cytotoxic effector cells in mice of the C57BL/6 strain that are relatively resistant to Friend, Moloney, and Rauscher (FMR) MuLV-induced leukemia, and in BALB/c mice that are relatively susceptible to leukemia induced by FMR MuLV. Generation of cytotoxicity requires in vivo administration of the virus followed by in vitro culture of lymphoid cells from virus-injected animals. Lymphoid cells from MuLV-resistant C57BL/6 donors develop high levels of specific cytotoxicity after secondary in vitro stimulation with syngeneic MuLV-induced tumor cells. Cells derived from these same donors, cultured in the absence of MuLV-induced tumor cells, fail to exhibit cytotoxicity. Secondary in vitro stimulation of lymphocytes from MuLV-susceptible BALB/c animals results not only in generation of cytotoxic reactivity against syngeneic MuLV-induced tumor cells but also induces apparently autoreactive effector cells capable of lysing other H-2d tumor cells as well as normal peritoneal cells bearing H-2d antigens. Moreover, generation of cytotoxicity by BALB/c lymphocytes occurs whether or not MuLV-induced tumor cells are included in the secondary culture system.
Immune Suppression in vivo with Antigen-Modified Syngeneic Cells
Authors: Sherr David H. , Cheung Nai-Kong V. , Heghinian Krikor M. , Benacerraf Baruj , Dorf Martin E.
Published: December 2022

Intravenous administration of syngeneic spleen cells coupled with the palmitoyl derivative of fowl γ-globulin (p-FγG) results in a profound state of FγG-specific tolerance in C57BL/6 mice. Administration of p-FγG coupled syngeneic cells specifically reduces both the primary and secondary hapten and carrier-specific PFC responses to TNP-FγG. Since the haptenic response is affected, the tolerance functions at the level of the FγG-specific helper T cell. As few as 103 p-FγG spleen cells carrying only 1 ng of p-FγG can induce tolerance. At least a 2-day-induction period is required. This nonresponsiveness is long lived, lasting over 120 days. Spleen cells from tolerized mice can transfer suppression to normal syngeneic recipients. Treatment of tolerant spleens with anti-Thy 1.2 antiserum + C eliminates the suppressor cell activity. In addition, thymocytes and purified splenic T cells from tolerized mice can transfer suppression to normal recipients. Thus, at least a component of this nonresponsiveness is mediated by suppressor T cells. The requirement of antigen association with cell membrane components and the general applicability of this method of inducing T cell nonresponsiveness are discussed.
The Structural Basis of a Hapten-Inhibitable κ-Chain Idiotype
Authors: Vrana Mark , Rudikoff Stuart , Potter Michael
Published: December 2022

The complete κ-chain variable region sequences from two murine myeloma proteins (U-61 and E-109) with binding specificity for β(2-1) fructofuranosyl moieties have been determined. The two κ-chains differ from each other by four amino acids. Earlier studies with recombinant molecules have assigned three different idiotypes to these two light chains. One of these determinants is hapten inhibitable and is presumed to be in or near the combining site, whereas the remaining two are not hapten inhibitable. Analysis of the four sequence differences found between these two chains in conjunction with a hypothetical three-dimensional model of the anti-fructan antibodies reveals that the hapten-inhibitable idiotype appears to be determined by the association of two amino acids found in complementarity-determining regions I and III. One of the nonhapten-inhibitable determinants could be caused by any of the three substitutions occurring in the framework portion of the variable region. The second of these determinants appears to result from the three-dimensional proximity of more than one framework amino acid.

A comparison of the U-61 and E-109 structures with partial sequences from several other light chains from fructofuranose-binding proteins revealed a total of eight differences occurring both in framework and complementarity-determining regions. The identification of Ser in two proteins at residue 30 and Asn at this position in three other proteins suggests that despite the great degree of homology exhibited by these light chains at least two germline genes would be required to code for these molecules.
Is Genetically Related Macrophage Factor (GRF) a Soluble Immune Response (Ir) Gene Product?
Authors: Erb Peter , Meier Barbara , Feldmann Marc
Published: December 2022

The possibility that the antigen-presenting “macrophages” interacting with helper cells either directly or via the intermediary action of a soluble factor consisting of Ia antigen and a fragment of immunogen, termed GRF (genetically related factor), are a site of Ir gene action was investigated by using the synthetic polypeptide antigen (T,G)-A—L. It was found that T cells from (responder × nonresponder) F1 mice were stimulated by responder “macrophages” or GRF derived from these cells but not by the nonresponder macrophages of GRF from these cells. This suggests that the defect in helper cell induction in nonresponders is at the level of the presenting cell and that the macrophage factor GRF is a soluble Ir gene product. This conclusion was supported by the observation that there was normal presenting cell and GRF function in nonresponders, mouse strains such as CBA that yield helper cells and helper factor with (T,G)-A—L and have defects elsewhere.
Two Migration Inhibitory Factors Differ in Density and Susceptibility to Neuraminidase and Proteinases
Authors: Remold Heinz G. , Mednis Alma D.
Published: December 2022

It has been shown that migration inhibitory factor (MIF) from guinea pigs can be separated into two subfractions. These are pH3-MIF (isoelectric point of 3.0 to 4.5 and apparent m.w. of 65,000) and pH5-MIF (isoelectric point of 5.0 to 5.5 and apparent m.w. of 25,000 to 40,000). Upon centrifugation of pH3-MIF in CsCl gradients, activity was found in the density range of 1.38 to 1.46 g/ml. pH5-MIF banded in CsCl gradients in fractions with a density ranging from 1.29 to 1.38 g/ml. Thus, pH3-MIF bands with glycoproteins having a high carbohydrate content. pH5-MIF has a density similar to pure proteins or to glycoprotein with low carbohydrate content. This interpretation is reinforced by the finding that pH3-MIF is neuraminidase sensitive, whereas pH5-MIF is neuraminidase resistant. Furthermore, the activity of pH5-MIF but not that of pH3-MIF was destroyed by treatment with trypsin. In contrast, chymotrypsin inactivated both MIF species. Thrombin and plasmin, on the other hand, did not inactivate pH5-MIF or pH3-MIF. The activity of pH5-MIF was also destroyed by incubation with peritoneal exudate cells, whereas the activity of pH3-MIF was unaltered after incubation with peritoneal exudate cells. Reaction of the peritoneal exudate cells with diisopropylfluorophosphate abolished their ability to inactivate pH5-MIF. Thus, inactivation of MIF is caused by a cell-associated serine protease(s) that may play a role in the regulation of the macrophage's response to one MIF species.
Studies of Congenitally Immunologic Mutant New Zealand Mice
Authors: Gershwin M. Eric , Castles James J. , Erickson Kent , Ahmed Aftab
Published: December 2022

Congenitally athymic (nude) mice on an NZB, NZW, and BALB/c background were produced by repetitive selective backcrossing. F'12 generation nude mice of these three strains were compared to their littermate nu/+ controls with respect to survival, histology, blood counts, splenic surface markers, response to mitogens, spontaneous plaque-forming cells, and appearance of naturally occurring thymocytotoxic antibodies (NTA). Under specific pathogen-free conditions, NZB nude mice survive less than 3 weeks, dying of a runting-like disease with infection by local normally noninvasive organisms. A contributing factor to this premature death is the relative absence of T cell progenitor populations in the NZB nude vs NZW nude or BALB/c nude groups. Furthermore, NZB nude mice have a significantly earlier appearance of NTA than nu/+ littermates and likewise appear to have heightened spontaneous polyclonal B cell responses against the haptens dansyl, nitroiodophenyl, trinitrophenyl, 2,4 dinitrophenyl, and sulfonate. It is suggested that NZB mice have several critical immunologic defects, including abnormalities of thymic epithelial cells, T cell differentiation pathways, and chronically polyclonal activated B cell populations. These defects interact to produce the clinical expression of autoimmunity.
Determination of the Number of Lytic Sites in Biconcave and Spheroid Erythrocyte Ghosts After Complement Lysis
Authors: Bauer Johann , Podack Eckhard R. , Valet Guenter
Published: December 2022

C5b-9 binding to erythrocyte membranes was quantitated with the aid of C7-depleted serum, reconstituted with 125-J-C7. The number of C5b-9 complexes/cell was correlated with the transformation of ghosts from the biconcave to the spheroid shape by the combination of flow cytometry and sucrose density gradient ultracentrifugation. At low complement levels the mean number of C5b-9 complexes ranged from 300 to 1200 complexes/erythrocyte ghost, and biconcave as well as spheroid ghosts were found simultaneously in the same sample. The separation of both types of ghosts by sucrose density gradient centrifugation showed that the irreversible transition from the biconcave to the spheroid form occurred at a critical value of 850 C5b-9 complexes/erythrocyte ghost. Ghosts bearing less than 850 C5b-9 complexes were biconcave, and resealed ghosts bearing more than 850 C5b-9 complexes were spheroid and leaky. The spheroid transition is abrupt; no intermediate forms could be detected. The spheroid transformation is interpreted as a direct interaction between the C5b-9 complexes and membrane proteins resulting in a disturbance of the cytoskeleton of the erythrocyte ghost followed by spheroidization.
The Activation of the Alternative Complement Pathway by Fetal Calf Serum and Mouse Thymocytes
Authors: Eisenberg Robert A. , Theofilopoulos Argyrios N. , Dixon F. J.
Published: December 2022

Mouse thymocytes activated the alternative complement pathway of mouse serum in the presence of heated fetal calf serum. The activation required C3 from the fetal calf serum but was independent of antibody either in the murine or bovine serum. No other murine cells tested, including erythrocytes, peripheral blood lymphocytes, lymph node cells, spleen cells, and various cultured cell lines, activated the alternative complement pathway as effectively as thymocytes. In addition, sera from species other than cows could not substitute for fetal calf serum. The C3 deposited on thymocytes was in the form of both C3b (immune adherence positive) and C3bi (conglutinable). We propose that the basis of activation in this system is the specific protection of bovine C3b on mouse thymocyte surface.
Insulin-Specific Murine Antibodies of Limited Heterogeneity
Authors: Kapp Judith A. , Strayer David S. , Robbins Paul F. , Perlmutter Roger M.
Published: December 2022

Immune responses by mice to porcine and bovine insulins are controlled by dominant genes linked to the H-2 gene complex. Mice bearing the H-2d haplotype respond to porcine insulin, whereas mice bearing other H-2 haplotypes do not; mice bearing the H-2b,d,v haplotypes respond to bovine insulin, whereas other strains do not. Analysis of serum antibody by isoelectric focusing demonstrates that antibodies specific for both porcine and bovine insulin are of restricted heterogeneity, suggesting that heterologous insulins stimulate a limited number of B cell clones. Banding patterns (spectrotypes) of sera from individuals of a given responder strain are remarkably uniform and these spectrotypic profiles are strain associated. In addition, phenotypic expression of spectrotypes is codominant in F1 hybrid mice and is controlled principally by allotype-linked genes. Analysis of cross-reactivity patterns demonstrates that all anti-porcine insulin antibodies bind bovine insulin and the majority of anti-bovine insulin antibodies bind porcine insulin. The minority of anti-bovine insulin antibodies that fails to bind porcine insulin may be specific for the A chain loop region of bovine insulin. These observations suggest that murine anti-insulin antibodies will provide a valuable model system for the identification of variable region genes that encode binding sites specific for protein molecules.
Increased Prostaglandin Production by Human Monocytes after Membrane Receptor Activation
Authors: Passwell Justen H. , Dayer Jean-Michel , Merler Ezio
Published: December 2022

Human monocytes in culture release small amounts of prostaglandin E (PGE) into the medium. Addition of Fc fragments of IgG to human monocyte monolayer cultures results in a marked increase in PGE release; Fab fragments, monomeric IgG, and human serum albumin have no effect. An IgG1 myeloma has no effect on PGE levels but addition of the heat aggregated protein results in a marked increase of PGE secretion. Exposure of the cells to Con A, which binds to a specific monocyte plasma membrane receptor, also results in a large increase in PGE release. The magnitude of the increase in PGE secretion produced by exposure of the monocytes to these ligands greatly exceeds the stimulation observed after the addition of antigen-activated mononuclear cell supernatants, zymosan, Sephadex beads, or endotoxin, to monocyte cultures. Prostaglandin E2 (PGE2) accounts for approximately 70% of the total prostaglandins released by stimulated cells.

After addition of Indomethacin to monocyte cultures, the stimulatory effects of the ligands on PGE release are inhibited. Addition of Con A to monocyte cultures results in an increased incorporation of [3H]-arachidonic acid into PGE2. These results suggest that this ligand stimulates synthesis as well as release of this prostaglandin.
A Role for Fatty Acid Composition of Complex Cellular Lipids in the Susceptibility of Tumor Cells to Humoral Immune Killing
Authors: Schlager Seymour I. , Ohanian Sarkis H.
Published: December 2022

Treatment of line-10 tumor cells with metabolic inhibitors (adriamycin, actinomycin D) or hormones (epinephrine, prednisolone) increases and decreases, respectively, the sensitivity of the cells to antibody-C mediated killing. Concomitantly, drug treatment inhibits the synthesis by the cells of specific complex lipids (cardiolipin, phosphatidyl choline, and triglycerides). Hormone treatment of these cells stimulates their synthesis of phosphatidyl choline and phosphatidyl serine and inhibits cardiolipin, phosphatidyl ethanolamine, and cholesteryl ester synthesis. Analysis of the newly synthesized fatty acids in the lipids extracted from drug-treated cells showed that the drug-induced sensitivity to antibody-C killing was accompanied by an increase in the amount of unsaturated fatty acids (C18:1, C18:2, and C18:3) in the cellular lipids. Cells washed free of drug and recultured in drug-free medium regained their resistance to antibody-C killing; these reverted cells had regained their ability to synthesize complex lipids and showed a fatty acid composition similar to that of untreated control cells. The effect of hormone treatment on the sensitivity of the cells to antibody-C killing could not be correlated with detectable changes in fatty acid synthesis. The results suggest that the drugs that increase the sensitivity of the cells to antibody-C killing may do so by altering the physical properties of the cell membrane through an effect on the synthesis of the fatty acid constituents of cellular lipids. Hormone treatment does not appear to affect the physical properties of the membrane by this mechanism.
Kinetic Analysis of Human Spontaneous Cell-Mediated Cytotoxicity
Authors: Pollack Sylvia B. , Emmons Sandra L.
Published: December 2022

We tested whether the kinetics of human spontaneous cell-mediated cytotoxicity (SCMC) fit the enzyme-substrate model for cellular cytotoxicity. As predicted by the model, the maximal rate of killing (Vmax) was proportional to the number of effector cells added, whereas the apparent affinity constant, K1/2, was a function of the source of effector cells, and remained unchanged when the number of effector cells was doubled. The linear initial velocity of lysis was maintained for only 90 min; thereafter, lysis proceeded at a lower rate. Thus the model strictly applies only during the initial phase.

Two lines of evidence suggested that NK cells that mediate SCMC function as recycling catalysts. Zone analysis of the log-dose relationship for SCMC indicated that effector cells are present in negligible amounts compared to the substrate. Although the rate of killing decreased with time, it appeared that each NK cell killed more than one target. When the same effector cell population was tested on two different targets, K562 and Chang, different K1/2 values were obtained. This result suggested that the two cell types are recognized differently, e.g., because they bear different determinants. Finally, we demonstrated formally that immune complexes competitively inhibit SCMC. If the enzyme-substrate analog holds, then FcR on NK cells are implicated either to be the “active site” for SCMC or to be closely linked to the active site.
Genetic Control of Immune Response to Sperm Whale Myoglobin in Mice
Authors: Okuda Kenji , Twining Sally S. , David Chella S. , Atassi M. Zouhair
Published: December 2022

The genetic control of T lymphocyte proliferative response to the five synthetic antigenic sites of myoglobin, two synthetic nonantigenic control peptides, and one “nonsense” peptide was determined in independent and recombinant strains of mice. In all the strains examined, the nonantigenic control peptides and the “nonsense” peptide did not invoke a response in myoglobin-primed mice. Further, when mice were not primed with whole myoglobin, no response was obtained with any of the antigenic sites. Haplotypes H-2d, H-2f, and H-2s are higher responders to sites 1 and 2, whereas haplotypes H-2d and H-2s are high responders to site 5. Response to site 3 may be controlled by a non-H-2-linked gene. Site 4 can stimulate H-2b and H-2k haplotypes that are nonresponders to the whole myoglobin. Studies with the recombinant strains suggested that Ir genes to sites 1 and 2 map in the I-A subregion and I-C subregion and were designated Ir-Mb-1,2(A) and Ir-Mb-1,2(C). Ir genes to sites 4 and 5 mapped only in the I-A subregion and were designated Ir-Mb-4(A) and Ir-Mb-5(A). These studies suggest that individual antigenic sites in a molecule are controlled by unique Ir genes.
Human Leukocyte Fc (IgG) Receptors: Quantitation and Affinity with Radiolabeled Affinity Cross-Linked Rabbit IgG
Authors: Alexander Elaine L. , Titus Julie A. , Segal David M.
Published: December 2022

Quantitative studies have been made of Fc receptors on human leukocytes derived from peripheral blood, thymus, tonsil, and spleen. The relative affinities and average numbers of receptors per cell were determined by measuring the binding of 125I-labeled, affinity cross-linked trimers of rabbit IgG to various populations of cells. In parallel, the sizes of receptor-bearing populations were determined by fluorescence microscopy. Fc receptors could be detected on leukocytes from peripheral blood and spleen, but not from tonsil or thymus. In the peripheral blood, the highest density of receptors was found on polymorphonuclear leukocytes; a subpopulation of lymphocytes had somewhat fewer receptors per cell, and circulating monocytes had the lowest receptor density. Among splenocytes, most of the receptors were found on myeloid cells and monocytes. In all populations, the affinity of Fc receptors for the trimer was about the same. At 0°C the average value for the association constant was 5 × 107 M-1.
Fractionation and Immunologic Assessment of KCl-Extracted Cardiac Antigens in Coxsackievirus B3 Virus-Induced Myocarditis
Authors: Paque Ronald E. , Straus David C. , Nealon Timothy J. , Gauntt Charles J.
Published: December 2022

Hypertonic salt extracts prepared from the heart tissues of adolescent CD-1 mice were fractionated on Sephadex G-100 columns. Two separate fractions were obtained. Fraction I, containing the antigenic immunoreactive activity, was able to inhibit the migration of CVB3-PPD immune mouse peritoneal exudate cells (IMPEC) as well as PEC from mice infected with CVB3 virus alone. Fraction II did not have antigenic activity as assessed by the agarose droplet cell migration inhibition assay. As controls, Fraction I prepared from the livers or spleens of CVB3-infected CD-1 mice was unable to inhibit the migration of CVB3 IMPEC. Unimmunized or “normal” mouse peritoneal exudate cells (NMPEC) were not inhibited by Fraction I. Antibodies prepared against Fractions I and II were unable to neutralize CVB3m virus in the plaque reduction test, and polyacrylamide gel analysis revealed multiple bands in 10% SDS gels.
The Genetics of PLT Response
Authors: DeWolf William C. , Carroll Peter G. , Mehta Cyrus R. , Martin Sharon L. , Yunis Edmond J.
Published: December 2022

PLT response is restricted by the HLA-D region. The present study was undertaken to help define the role of HLA-DRw in PLT restimulation. Haplotype-primed intrafamily PLT cells were made against specificities HLA-Drw1, HLA-DRw3, and HLA-DRw7; each PLT was then restimulated with cells from a 35-member unrelated panel. Restimulation values for each PLT were subjected to bimodal clustering analysis. In addition, blocking experiments were performed with other intrafamily and homozygous typing cell PLT after preincubation with B cell alloantisera. The results show a high correlation (0.881 ≤ r ≤ 1.00) between the HLA-DRw specificity of the priming haplotype and the HLA-DRw specificity of unrelated panel cells that restimulate in PLT. When stimulating cells were absorbed with the corresponding DRw alloantisera or p29,34 heteroantiserum (against B cell specific antigens), PLT restimulation was significantly blocked. However, the PLT cells treated with antisera showed no effect. The results strongly suggest that HLA-DRw is the principal PLT-stimulating determinant.
Glucocorticoids Inhibit Trypsin-Induced DNA Release from Phytohemagglutinin-Stimulated Blood Lymphocytes
Authors: Distelhorst Clark W. , Rogers John C.
Published: December 2022

Trypsin-induced DNA release from phytohemagglutinin (PHA)-stimulated human lymphocytes is inhibited by different glucocorticosteroid compounds at low pharmacologic concentrations, in a dose-dependent manner, and in order of the known anti-inflammatory potency of the different preparations, In contrast, PHA-stimulated cell growth is 100- to 1000-fold less sensitive to inhibition by the same glucocorticoids. Nonglucocorticoid steroids have little effect on either DNA release or cell growth except at high concentrations. Inhibition of DNA release appears to be mediated through glucocorticoid receptors since progesterone, which is ineffective alone, competitively inhibits the effect of dexamethasone. The glucocorticoid effect on DNA release is tightly coupled to the initial, PHA-induced stimulus. Glucocorticoids are maximally effective when added to cultures 1 hr before PHA. When added 6 hr after PHA, their effect is minimal or absent, even though they are then continuously present until DNA release is measured 5 days later. Lymphocytes from certain donors in these studies were resistant to glucocorticoids; these individuals all had allergies, including asthma, allergic rhinitis, and bee sting hypersensitivity.
Complement-Mediated Solubilization of Immune Precipitates Prepared with Antibodies of Different Avidity
Authors: Stassen Frans L. , Beek Cor J.
Published: December 2022

Complement-mediated solubilization of immune precipitates prepared with HSA and rat IgG anti-HSA has been quantitatively analyzed. Early and late IgG anti-HSA antibodies were obtained 27 and 49 days after immunization, respectively. Immune precipitates prepared with early IgG anti-HSA were solubilized by rat serum to a larger extent than complexes prepared with late IgG anti-HSA. The affinities for HSA of the early and late antibodies were not significantly different. The quantitative differences in solubilization were neither due to differences in the Ab/Ag ratios of the immune precipitates, nor appeared to be brought about by changes in the distribution of the antibodies over the IgG subclasses. The avidity of the late IgG anti-HSA antibodies was higher than the avidity of the early IgG antibodies. Presumably, the avidity of the antibodies greatly affected the complement-mediated solubilization of the immune precipitates. In addition, the solubilization was found to be dependent on the conditions employed to prepare the immune precipitates.
Role of Surface IgM and IgD in the Functional Differentiation of Human B Lymphocytes: Effect of Papain Treatment
Authors: Dosch Hans-Michael , Kwong Shirley , Tsui Florence , Zimmerman Barry , Gelfand Erwin W.
Published: December 2022

Short-term treatment of normal human B lymphocytes with low concentrations of papain resulted in selective and reversible removal of sIgD determinants, whereas HLA and Ia-like antigens, sIgM as well as receptors for E, C3, and FcIgG were unaffected. When studied for their capacity to generate antigen-specific direct PFC, papain-treated (δ-) B cells were highly sensitive to inactivation by even low concentrations of antigen. In addition, these cells were impaired in their ability to cooperate normally with T-helper cells or their humoral product(s).
C3e: An Acidic Fragment of Human C3 with Leukocytosis-Inducing Activity
Authors: Ghebrehiwet Berhane , Müller-Eberhard Hans J.
Published: December 2022

We have isolated a leukocytosis-inducing peptide of degraded human C3 that has the electrophoretic behavior of prealbumin and a m.w. of 10,000 to 12,000. The peptide has been designated C3e. It was generated by two methods: Incubation of isolated C3 with 2% (w/w) trypsin for 120 min at 37°C, or incubation of serum at 37°C for 5 days under sterile conditions. C3e preparations obtained by the two methods were virtually indistinguishable with respect to molecular size, amino acid composition, and immunochemical properties. The electrophoretic mobility of C3e at pH 8.6 is -7.5 × 10-5 cm2 V-1 sec-1. Anti-C3e reacts with C3e, C3, C3b, and C3c, but not with C3a or C3d. Since the β-chain of C3 is not affected by the liberation of C3e, it is concluded that C3e arises from cleavage of the portion of the α-chain that is part of the C3c fragment. Purified C3e was found to mobilize leukocytes from bone marrow upon perfusion of isolated rat femur. Intravenous injection of C3e into rabbits caused a 2- to 3-fold increase in the number of circulating leukocytes after 60 min which lasted 2 hr. Leukocytosis was preceded by mild leukopenia. Leukocytosis-inducing activity was abrogated by treating C3e preparations with solid phase anti-C3. Incubation of heparinized rabbit blood with radioiodinated C3e resulted in preferential binding of C3e to polymorphonuclear leukocytes. Intradermal injection of C3e into rabbits evoked increased vascular permeability after 15 to 30 min. The evidence indicates that C3e is a fragment of C3 with biologic activity distinct from that of other C3 fragments.
Functional Studies of Fc Receptor-Bearing Human Lymphocytes: Effect of Treatment with Proteolytic Enzymes
Authors: Perussia Bice , Trinchieri Giorgio , Cerottini Jean-Charles
Published: December 2022

In order to analyze the role that Fc receptors (FcR) play in spontaneous cell-mediated cytotoxicity (SLMC), lymphocytes from single donors were tested in parallel for their ability to form rosettes with IgG antibody-coated erythrocytes (EA), and to mediate antibody-dependent cytotoxicity (ADCC) and SLMC. Experimental conditions were chosen in which the expression of FcR at the cell surface is altered (i.e., after enzymatic treatment or reaction with immune complexes). Trypsintreated lymphocytes still expressed FcR and were able to mediate ADCC, but SLMC was completely abolished. Pronase treatment, on the contrary, destroyed FcR and both ADCC and SLMC. When the enzyme-treated cells were cultured for up to 48 hr FcR was reexpressed at the cell surface, and both lytic activities were regained. SLMC from B cell-deprived trypsin-treated cells was recovered in culture in spite of the presence of F(ab′)2 anti-human F(ab′)2 antibodies in the culture medium and throughout the test. Moreover, incubation of the trypsintreated cells in medium containing autologous serum could not reconstitute SLMC. When lymphocytes were reacted with immune complexes, a modulation of both ADCC and SLMC was observed. In this case, the lytic activities could not be recovered even after incubation at 37°C. A partial inhibition of the modulation of FcR was observed when the cells were reacted with immune complexes in the absence of Ca++ ion. The possible existence of two different mechanisms operating in ADCC and SLMC is discussed.
Further Evidence for the Antibody Nature of C3 Nephritic Factor (C3NeF)
Authors: Daha Mohamed R. , van Es Leendert A.
Published: December 2022

C3 nephritic factor (C3NeF), found in the sera of some patients with membranoproliferative glomerulonephritis, has been shown to be composed of two heavy and two light chains, like IgG; in addition it shares antigenic determinants with IgG. Purified C3NeF binds to the amplification convertase of complement, C3b,Bb, and thereby prevents decay of its C3-cleaving potential. The capability of C3NeF to bind to C3b,Bb was used as a means for purifying C3NeF to homogeneity. The investigation described in this report suggests that binding of C3NeF to C3b,Bb occurs via the Fab portion of the molecule. Pepsin treatment of eight C3NeF preparations resulted in an average loss of 76% of C3NeF functional activity. Papain treatment induced a loss of approximately 90%. The decrease in functional activity could be attributed to the accelerated rate of dissociation of 125I-F(ab′)2 and 125I-Fab fragments from stabilized cell-bound C3b,Bb. The dissociation rate of 125I-F(ab′)2 from C3b,Bb was comparable with the decay of the functional activity of C3b,Bb stabilized by F(ab′)2 or Fab fragments of C3NeF. Although these results suggest that the stabilizing activity of C3NeF is mediated by the Fab portion of the molecule, it was found that the Fc portion also contributes to its functional activity.
Chemical Cross-Linking of Ia Alloantigen α and β Chains with Dimethyl 3,3′-Dithiobispropionimidate
Authors: Dancey George F. , Cutler John , Schwartz Benjamin D.
Published: December 2022

We have examined the polypeptide chain composition of membrane-bound and detergent-solubilized Ia antigens using the chemical cross-linking reagent dimethyl 3,3′-dithiobispropionimidate (DTBP). Products of the I-E/C subregion of the major histocompatibility complex, which were solubilized from spleen cells with the detergent NP-40 and partially purified by affinity chromatography on lentil lectin-agarose, could be almost completely cross-linked by DTBP. Thus, the characteristic 33,000 m.w. (α) and 28,000 (β) polypeptide chains seen on sodium dodecylsulfate polyacrylamide gels disappeared and a major new species of 60,000 m.w. appeared after cross-linking. When isolated and reduced with 2-mercaptoethanol, the 60,000 m.w. peak was found to be comprised of α and β chains. Similar results were obtained when I-E/C, as well as I-A, α and β chains were cross-linked on the cell surface. These data demonstrate that the α and β chains of the Ia antigens exist primarily in the form of a dimer both in detergent solution and in situ.
T Cell Recognition in the Mixed Lymphocyte Response
Authors: Ahmann Gerald B. , Nadler Paul I. , Birnkrant Alan , Hodes Richard J.
Published: December 2022

The ability of subpopulations of murine spleen cells to stimulate a mixed lymphocyte response (MLR) was studied. It was found that T cells (nylon-nonadherent spleen cells) and B cells [G-10 passed and treated with rabbit anti-mouse brain serum (RAMB) and complement (C)] were poor stimulators of an MLR. In contrast, whole spleen cells or B cells plus adherent cells (RAMB +C-treated spleen cells) produced good stimulation. However, a non-T, radiation-resistant splenic adherent cell (SAC) population was up to 20 to 50 times more efficient as a stimulator of an MLR on a per cell basis than an unseparated spleen population. These SAC were shown to express Ia determinants encoded by genes in I-A and I-E/C. These results suggest that Ia+ SAC may be the predominant stimulating cells in spleen cell populations, and the preferential target for T cell recognition in cell interaction events.
The Relationship Between Surface Immunoglobulin Isotype and Immune Function of Murine B Lymphocytes
Authors: Zan-Bar I. , Strober S. , Vitetta E. S.
Published: December 2022

We developed a quantitative procedure to examine the ability of memory B cells to give rise to AFC, and to more memory B cells (propagation of memory) after antigenic stimulation. Sublethally irradiated (BALB/c × C57BL/Ka)F1 mice were given an excess of BSA-primed T cells, graded numbers of DNP-primed B cells, and challenged with DNP-BSA in saline. The anti-DNP response of these adoptive (intermediate) hosts measures the production of AFC. After a rest period of 8 to 12 weeks, spleen cells from these intermediate hosts were transferred to sublethally irradiated final hosts. The latter animals were also given an excess of primed T cells and challenged with DNP-BSA. The anti-DNP response of the final hosts measures the ability of memory B cells to propagate memory.

The relationship between the expression of surface immunoglobulin isotype (IgM, IgD, IgG), and the ability to produce AFC or propagate memory was studied by purifying populations of IgM+, IgD+, and IgG+ memory B cells on the FACS. The experimental results show that IgM+, IgD+, and IgG+ cells give rise to AFC, but that only the IgD+ subset can propagate memory. This suggests that memory B cells lacking surface IgD undergo terminal differentiation after antigenic stimulation, but that those bearing IgD are capable of self-renewal.
Induction of Allograft Tolerance after Total Lymphoid Irradiation (TLI): Development of Suppressor Cells of the Mixed Leukocyte Reaction (MLR)
Authors: Slavin S. , Strober S.
Published: December 2022

We searched for the presence of suppressor cells of the MLR in C57BL/Ka → BALB/c chimeras. The chimeras were made with total lymphoid irradiation (TLI) and marrow transplantation. Spleen cells from the old chimeras inhibited the MLR of BALB/c responder cells against C57BL/Ka stimulator cells. Inhibition was specific for the stimulator cells, since no effect on the MLR was observed with C3H or BALB.C3H stimulator cells. Maximal inhibition was achieved when the responder cells in the MLR shared the H-2 haplotype of the chimeric recipient. Spleen cells obtained from chimeras young 30 to 40 days after BM transplantation inhibited the MLR nonspecifically, since similar marked inhibition was observed regardless of the H-2 haplotype of the responder or stimulator cells. The finding of antigen-specific and nonspecific suppressor cells is similar to that observed in mice rendered tolerant to bovine serum albumin after treatment with TLI.
The Heterogeneous Distribution of 5′-Nucleotidase Among Rabbit Lymphocytes
Authors: Gregory Stephen H. , Kern Milton
Published: December 2022

Assay of 5′-nucleotidase activity revealed a nonuni-form distribution of enzyme activity among rabbit lymphoid tissues. Although 5′-nucleotidase activity was readily observed in rabbit spleen cells, it was essentially undetectable in cells derived from thymus, blood, bone marrow, and appendix. Popliteal lymph node cells, on the other hand, exhibited approximately 6% of the specific activity observed in spleen. Cytochemical reactions carried out on splenocyte smears showed that only 20% of the spleen cells in the total population possessed demonstrable 5′-nucleotidase activity. The results of cell separation studies indicated that the bulk of the activity was not principally associated with either T cells, B cells, or macrophage cells. However, the association of some enzyme activity with subpopulations of these cell types could not be excluded by this kind of analysis. Autoradiographic studies indicated that 5′-nucleotidase did not afford a selective advantage to those splenocytes possessing the enzyme inasmuch as both enzyme-bearing and enzyme-nonbearing cells, when present in the same population, readily incorporated label derived from extracellular [3H]adenosine-labeled 5′-AMP. Thymus cells, which did not contain a 5′-nucleotidase-bearing population were unable to utilize extracellular 5′-AMP.

The distribution of two other plasma membrane-associated enzymes was also explored. Whereas nucleotide pyrophosphatase was more uniformly distributed, the distribution of phosphodiesterase I was quite heterogeneous yet distinguishable from that observed for 5′-nucleotidase.
Immunoregulation of Localized and Disseminated Murine Myeloma: Antigen-Specific Regulation of Mopc-315 Stem Cell Proliferation and Secretory Cell Differentiation
Authors: Rohrer James W. , Lynch Richard G.
Published: December 2022

Tumor development, MOPC-315 stem cells, and M315-secretory cells were quantitated in carrier-primed BALB/c mice that had been challenged subcutaneously or i.v. with mixtures of TNP-carrier and TNP-binding MOPC-315 cells. We observed that tumor incidence, myeloma stem cells, and secretory myeloma cells were: i) suppressed in mice in whom carrier-specific suppressor T cells had previously been induced and ii) initially enhanced in mice with carrier-specific helper T cells. The early enhancement in mice with carrier-specific helper T cells was followed by progressively declining myeloma stem cell frequencies and regression of established tumors. These studies demonstrate that T cell-derived immunoregulators of host origin can be focused onto localized and disseminated malignant B cells and specifically regulate the expansion and differentiation of the neoplastic clone.
Suppressor Activity of T Lymphocytes from Infants Assessed by Co-Culture with Unfractionated Adult Lymphocytes in the Pokeweed Mitogen System
Authors: Miyawaki Toshio , Seki Hidetoshi , Kubo Minoru , Taniguchi Noboru
Published: December 2022

T lymphocytes were isolated by E rosette sedimentation from cord blood lymphocytes and from the peripheral blood lymphocytes of healthy children of various ages. Fc(IgG) receptor-bearing T lymphocytes were enumerated by rosette formation with ox erythrocytes coated by IgG fraction of rabbit antisera. Relative proportions of Fc(IgG) receptor-bearing T cells in T cell-enriched populations of cord blood and newborn infants were higher than those in later life and decreased to near healthy adult levels by 3 months of age. Only a few of lymphocytes in cord blood were differentiated into immunoglobulin(Ig)-producing cells in vitro by poke-weed mitogen-(PWM) stimulation. The generation of Ig-producing cells in peripheral blood lymphocytes increased with age and reached about half the adult mean at 3 years of age.

The suppressor activity of T lymphocytes were evaluated by co-culture with unfractionated adult lymphocytes on PWM-system. The PWM-induced differentiation of adult B lymphocytes into Ig-producing cells was markedly suppressed by the addition of T lymphocytes from cord blood. The suppressor activity of cord T lymphocytes was dose dependent to some extent and appeared to be equally effective on the generation of Ig-producing cells of three major classes, IgG, IgA, and IgM. Such suppressor activity of T lymphocytes was observed throughout infancy and gradually decreased with age. At 2 years of age or later, the suppressor activity of T lymphocytes, as evaluated by co-culture with adult lymphocytes, was not consistently demonstrated.
Lymphocytes Surface Membrane Immunoglobulin in Myeloma
Authors: Gebel Howard M. , Hoover Richard G. , Lynch Richard G.
Published: December 2022

MOPC-315 is a BALB/c plasmacytoma that produces an IgA anti-TNP antibody (M315). 315/+ is a line derived from MOPC-315 and differentiates during in vivo growth from M315-producing, nonsecreting, lymphocytoid cells to large M315-secreting plasmacytoid cells. Studies were conducted to determine whether the small M315-bearing mononuclear cells, which account for more than one-third of the circulating mononuclear cells in mice with 315/+ tumors, were myeloma cells or host lymphocytes. Experiments conducted in CBF1 mice demonstrated that the circulating M315-bearing cells were of F1, rather than tumor, origin. Serologic and ultrastructural studies demonstrated that the cells were post-thymic T lymphocytes. After proteolytic removal, surface M315 was reexpressed in vitro by 315/+ cells, but not by T cells. M315-bearing T cells accounted for 0 to 4% of circulating mononuclear cells in mice with 315/P, a variant of MOPC-315 in which all cells synthesize M315, but only 2% of the cells are secretory. There was no obvious relationship between the frequency of M315-bearing lymphocytes and development of the humoral immunodeficiency that accompanies myeloma. These findings 1) identify an association between high levels of M315 secretion and development of M315-bearing T cells; 2) favor the view that M315 is acquired, rather than produced by host T cells; 3) raise the possibility that T cells with IgA-Fc receptors may be increased in mice with 315/+ tumors; and 4) are discussed relative to the mechanisms that have been proposed in the literature to account for the occurrence and significance of circulating, M-component-bearing lymphoid cells in murine and human myeloma.
An Immunologic Approach for the Detection of Tumor Cells in the Peripheral Blood of Patients with Malignant Lymphoma; Implications for the Diagnosis of Minimal Disease
Authors: Ligler Frances S. , Vitetta Ellen S. , Smith R. Graham , Himes Joyce B. , Uhr Jonathan W. , Frenkel Eugene P. , Kettman John R.
Published: December 2022

Lymphocytes were isolated from infiltrated lymph nodes and blood of eight patients with malignant lymphocytic lymphoma. All patients had normal numbers of circulating lymphocytes with normal morphology. None of the patients had serum paraproteins. The isolated lymphocytes from patients and controls were stained with affinity purified antibody specific for κ- or λ-light chains and analyzed by using a fluorescence activated cell sorter. Cells from infiltrated nodes stained for only one light chain type at high levels of fluorescence intensity. An increase in the number of brightly stained cells bearing the light chain type found in the involved node was readily detected in the peripheral blood of all but one of the patients with lymphoma. Similar changes in the ratio of κ+- to λ+-cells were not observed in 41 samples from normal individuals and patients with other diseases.
In Vitro Immune Response of Human Peripheral Lymphocytes
Authors: Hirano Toshio , Kishimoto Tadamitsu , Kuritani Taro , Muraguchi Atsushi , Yamamura Yuichi , Ralph Peter , Good Robert A.
Published: December 2022

An antiserum to a human T leukemia cell line, HSB, absorbed with the homologous B cell line, SB, reacted with three of five T cell lines, none of five B cell lines, and none of three other human hematopoietic cell lines. The absorbed antiserum (αHSB) killed 23 to 56% normal peripheral blood lymphocyte (PBL) T cells, depending on the donor, but not B cells. Treatment of purified PBL T cells with αHSB and complement (C) had no specific effect on their mitogenic responses of PHA and Con A mitogens, but partially inhibited MLC reactions. αHSB and C had no effect on helper T cells or B lymphocytes in pokeweed mitogen (PWM)-induced immunoglobulin (Ig) production. When continuously present in the culture, αHSB at 1:400 dilution almost completely inhibited IgG and IgM production induced by PWM. T cells pretreated with αHSB for 3 days and washed were capable of inhibiting Ig production by normal PBL at ratios less than 1:100. In contrast, αHSB-treated B cells had no effect on PWM induction of Ig. The suppressor T cells induced by αHSB carried an antigen recognized by the antiserum on their surface as shown by removal of suppression by pretreatment with αHSB and C. αHSB-treated T cells secreted a soluble factor suppressing Ig production by PWM-stimulated PBL.