bluuebunny/crossref_metadata_2025 huggingface dataset:
Cell-Mediated Immunity to Friend Virus-Induced Leukemia
Authors: Ting Chou-Chik , Bonnard Guy D.
Published: December 2022

Primary and secondary cell-mediated cytotoxic responses to FBL-3 cells, a syngeneic Friend virus-induced leukemia in C57BL/6 mice, could be generated by in vitro techniques as tested by the 125IUdR release assay. The specificity of the cytotoxic reactions appeared to be directed against the Friend type-specific antigen and the FMR (Friend, Moloney, Rauscher) antigen which were also the major antigens for transplantation immunity to FBL-3. In comparison to the primary cytotoxic response, the secondary cytotoxic response was accelerated (detected at an earlier time after sensitization), enhanced (gave much higher levels of cytotoxicity), was also longer lasting, and could be induced by a wide dose range of tumor cells. The secondary response could only be induced with lymphocytes obtained from regressors that were resistant to FBL-3 challenge; lymphocytes from mice with progressive tumor growth had no detectable secondary response. It was found that both induction phase and the effector phase of cytotoxic responses were T cell dependent. The characteristics of these reactions were thus very similar to those obtained with in vivo immunization or challenge, providing a good correlation with in vivo tumor immunity.
Regulatory Serum Lipoproteins: Regulation of Lymphocyte Stimulation by a Species of Low Density Lipoprotein
Authors: Curtiss Linda K. , Edgington Thomas S.
Published: December 2022

Low density lipoproteins (LDL) containing apolipoprotein B were separated from 15 fresh normal human serum pools by three independent isolation methods including sequential ultracentrifugal flotation, affinity chromatography, and polyanion precipitation. A discrete subpopulation of LDL (LDL-In) was isolated which possessed comparable inhibitory activity for PHA, PWM, and allogenic cell stimulated human lymphocytes in vitro at concentrations of 1 to 10 µg protein/1 × 105 lymphocytes/0.25 ml culture. LDL-In was characterized by a mean buoyant density of 1.055 g/ml in KBr, a m.w. of 2 to 3 × 106 daltons and a composition of 20 to 25% protein and 75 to 80% lipid with β electrophoretic mobility. The biologic activity of LDL-In was non-cytotoxic, independent of mitogen concentration, and dependent upon the concentration of serum in the culture assay.

The effect was temporally dependent requiring approximately 24 hr for induction of a stable suppressed state. Suppression was reversible with shorter periods of exposure to LDL-In. LDL-In did not inhibit lymphocytes at periods greater than 19 hr after stimulation, suggesting that LDL-In may influence metabolic events associated with the inductive phase of lymphocyte activation by lectins and allogeneic cells. LDL-In was clearly distinguishable from T lymphocyte E rosette inhibitory factor since it did not influence E rosette function of lymphocytes. The physicochemical and biologic properties of LDL-In clearly distinguish this regulatory lipoprotein from previously described immunoregulatory factors.
Effect of Murine Cytomegalovirus on the in Vitro Responses of T and B Cells to Mitogens
Authors: Selgrade Maryjane K. , Ahmed Aftab , Sell Kenneth W. , Gershwin M. Eric , Steinberg Alfred D.
Published: December 2022

Both in vitro and in vivo murine cytomegalovirus (MCMV) infection depressed the responses of lymphocytes to both B and T cell mitogens. The possibilities that macrophages or nonspecific T cell inhibition of B cells might account for the depressed responses were eliminated. In vitro data suggested that B cell responses are more susceptible to this depression than T cell responses. The possibility that the depression of T cell responses is not a direct effect of viral infection of lymphocytes is discussed.

To investigate further the interaction between B and T lymphocytes and MCMV, mice with B and T cell deficiences were studied. A comparison of the susceptibility of athymic Nu/Nu mice and T cell competent Nu/+ littermates to MCMV showed that the LD50 for Nu/Nu mice is 10-fold lower than that for Nu/+ mice, but Nu/+ mice given an LD50 of virus died much sooner after infection than Nu/Nu mice given an LD50. Pathogenic mechanisms responsible for death may be different in these two groups of mice. Similarly the MCMV LD50 for B cell-deficient mice (treated with goat anti-mouse IgM serum) was 10-fold lower than the LD50 for mice treated with normal goat serum, but given an LD50 of virus, the latter died sooner after infection than the former. In contrast, there was little difference between the LD50 or time of death after MCMV infection of CBA × DBA F1 male mice (which are deficient in their response to thymic independent antigens) and their normal littermates, the CBA × DBA F1 female mice.
Studies on the Terminal Stages of Antibody-Complement Mediated Killing of a Tumor Cell
Authors: Boyle Michael D. P. , Ohanian Sarkis H. , Borsos Tibor
Published: December 2022

A comparison of the events leading from T* to dead cells and E* to E ghost and hemoglobin was made. Transformation of T* to dead cells was inhibitable by EDTA, NaF, histamine, DMSO, cytochalasin B and 3′5′ cAMP. Transformation of E* to E ghost and hemoglobin was only inhibited by EDTA.
Effect of Six Weekly Transfusions on Canine Marrow Grafts: Tests for Sensitization and Abrogation of Sensitization by Procarbazine and Antithymocyte Serum
Authors: Weiden Paul L. , Storb Rainer , Slichter Sherrill , Warren Reed P. , Sale George E.
Published: December 2022

We have previously shown that blood transfusions can immunize a dog and lead to rejection of a subsequent marrow graft despite lethal total body irradiation (TBI). Sensitization to histocompatibility antigens induced by two prior transfusions of whole blood could be overcome by a regimen of procarbazine and anti-thymocyte serum (ATS) preceding TBI. The current study investigated a) whether this regimen could abrogate sensitization induced by six weekly transfusions given from days -50 to -15 preceding a marrow graft, and b) whether platelet survival studies and two in vitro tests of immunity could predict marrow graft rejection.

All donor-recipient pairs were histoincompatible, unrelated, and of different breed. Twenty-two recipients received platelet concentrate transfusions and eight received whole blood transfusions. Recipients were given 1200 R TBI and a graft of marrow and peripheral blood leukocytes from the transfusion donor on day 0. Three of 15 recipients (20%) given procarbazine, 12.5 mg/kg i.v. on days -8, -6, and -4, and ATS, 0.6 ml/kg subcutaneously on days -7, -5 and -3, rejected their grafts, whereas 11 of 15 dogs (73%) not given procarbazine and ATS rejected their grafts (p < 0.01).

Serum lymphocytotoxic antibodies, peripheral leukocyte migration inhibition, and in vivo donor platelet reccvery and survival were studied in those recipients receiving six weekly transfusions and in 18 other recipients receiving a single donor transfusion 3 months before marrow grafting. No significant correlation was found among these in vitro and in vivo tests of sensitization. Sensitization to marrow grafts was not reliably detected by the presence of cytotoxic antibodies or leukocyte migration inhibition. Platelet survival, however, was positively correlated with the results of marrow grafting in 12 of 15 (80%) evaluable recipients (p ∼ 0.15).

This study confirms, with a different immunization schedule, that procarbazine and ATS can abrogate sensitization to histocompatibility antigens induced by multiple prior transfusions in histoincompatible, unrelated canine donor-recipient pairs and thus permit successful engraftment of donor marrow.
Inhibition of Antigen-Induced B Lymphocyte Activation in Vitro by Cytochalasin B
Authors: Roberts Robert L. , La Via Mariano F.
Published: December 2022

Cytochalasin B (CB), a fungal metabolite which disrupts microfilaments. and will inhibit capping and other events requiring membrane movement, suppressed the production of antibody-forming cells (AFC) in both mouse whole spleen cultures immunized with sheep erythrocytes (SRBC) and in mouse B lymphocyte cultures immunized with the thymic independent antigen DNP-Ficoll (DF). CB at a concentration of 1 µg/ml inhibited the AFC response by more than 90% in spleen cell cultures immunized with SRBC. This inhibition was completely reversible by removal of CB up to 24 hr after the start of culture. Spleen cells cultured in the presence of CB for the first 48 to 72 hr had a decreased AFC response similar to that of cultures in which SRBC had been withheld for the same period of time. Incubating whole spleen cell or B lymphocyte cultures immunized with DF for as short as 6 hr decreased the AFC response more than 60%. Antibody secretion, cell viability, and antigenicity of the SRBC and DF were not affected by CB.

The results of these experiments favor the concept that movement of surface receptors is necessary in activating lymphocytes to differentiate into AFC. The differential response to CB observed in SRBC and DF stimulated cultures makes the technique employed a useful tool to study membrane events occurring between antigen interaction with surface receptor and the initiation of differentiative events.
Diversity among Rabbit Antibody Light Chain Amino Terminal Sequences
Authors: Cannon L. Edward , Margolies Michael N. , Strosberg A. Donny , Chen Francis W. , Newell John , Haber Edgar
Published: December 2022

The amino terminal (positions 0 to 20) amino acid sequences of 22 rabbit antibody light chains are reported and compared with 44 amino terminal sequences previously described in the literature. Rabbit κ light chains demonstrate three different amino terminal chain lengths and considerable variability at the amino terminal end. In relating amino terminal sequence to b locus allotype, allotype-specific residues (amino acid residues unique to all light chains having a given allotype) were not identified, although certain residue alternatives were unique to some of the b5 and b9 light chains.

There was no correlation of antibody specificity with amino terminal sequence. In the most striking example, identical amino terminal sequences were associated with two different carbohydrate specificities (type VIII pneumococcal polysaccharide and streptococcal group A carbohydrate) and an aminophenyltrimethylammonium specificity. An obligatory association of amino terminal sequence with complete variable region framework sequences was not observed. Thus, it is not always possible to predict the framework homology among a given pair of light chains by examination of their amino terminal sequences.

The limited diversity seen among the variable regions of murine myelomas that have the same antigen-binding properties is not found in the light chains of specifically elicited rabbit antibodies.
Humoral Immunity in Experimental Syphilis
Authors: Bishop Nancy H. , Miller James N.
Published: December 2022

Evidence for a humoral mechanism in immunity to experimental syphilis was provided by the demonstration of immune rabbit serum factor(s) capable of inactivating virulent Treponema pallidum, Nichols strain, in an in vitro-in vivo neutralization test. After intratesticular infection, rabbits were bled periodically and their resistance to reinfection was determined by challenge with T. pallidum. The results of challenge showed that resistance to reinfection begins to develop by 11 days after infection, becomes complete by 3 months, and persists for at least 2 years. In the neutralization test, a mixture of treponemal suspension and serum from the infected animals was incubated anaerobically at 34°C and the virulence of the treponemes was determined by intradermal inoculation into normal rabbits. Complete inactivation of treponemes by immune serum required heat-stable and heat-labile (56°C, 30 min) serum components and 16 hr of incubation, and was accelerated by pre-incubation of the treponemes for 4 hr with nonimmune serum but not by 100 µg/ml of added lysozyme. Serum-neutralizing activity, first demonstrable 1 month postinfection, was quantitated by a neutralizing endpoint (NEP). A relatively close quantitative correlation was shown between the development of resistance to symptomatic reinfection and the appearance and persistence of both TPI antibody and neutralizing serum factor(s). The nature of the serum factor(s), the mechanism of treponemal inactivation, and the application of the test in assessing the immune status are discussed.
Studies on Glycopeptide Released by Trypsin from Sheep Erythrocytes
Authors: Kitao Takeshi , Takeshita Masazumi , Hattori Kenichi
Published: December 2022

Pretreatment of sheep erythrocytes with trypsin abolishes their specific binding and rosette formation with human T lymphocytes. A glycopeptide containing sialic acid is released from the intact sheep erythrocytes by incubation with trypsin and purified. This glycopeptide contains activity that can be bound to T lymphocytes and produces inhibition of rosette formation. This component with a m.w. of about 10,000 contains galactose, acetylglucosamine, acetylgalactosamine, sialic acid, and serine. These results suggest that the glycopeptide released by trypsin treatment may contain the site of the T cell receptor of sheep erythrocytes.
Isolation and Partial Characterization of AgF-1:A Rat Lymphocyte Membrane Antigen
Authors: Williams Phillip B. C. , DeWitt Charles W.
Published: December 2022

A genetically defined, serologically identified antigen of the rat lymphocyte membrane (AgF-1) has been isolated. Viable spleen and lymph node cells, prepared by Ficoll-Hypaque density centrifugation from Fischer rats, were radioiodinated with soluble lactoperoxidase. Extracts obtained with Nonidet P-40 were shown to contain numerous radiolabeled proteins including cell-surface globulin. AgF-1 was isolated from these extracts by precipitation with a highly specific alloantibody in conjunction with xenospecific anti-globulin antibody and polyethylene glycol (PEG). The use of PEG greatly increased the efficiency of the double antibody technique.

The putative antigenic peak was eluted from sodium dodecyl sulfate polyacrylamide gels (SDS-PAGE) and specific antigenic activity was recovered. Removal of the SDS from these eluates was achieved by equilibration with urea and passage over an anion exchange resin. Renaturation, as evidenced by specific inhibition of complement-mediated cytotoxicity, occurred upon the removal of urea by dialysis. The m.w. of the purified antigen was estimated to be 35 to 40,000 daltons by SDS-PAGE and was unaffected by reduction with 2-mercaptoethanol. Amino acid composition was roughly similar to those reported for the major histocompatibility antigens of the rat and other species.
Antibody to Murine Leukemia Virus: Genetic Control Linked to the H-2 Locus in PL Mice
Authors: Nowinski Robert C. , Doyle Theresa
Published: December 2022

Mice of a wide variety of inbred strains naturally produce antibody against endogenous ecotropic leukemia viruses (MuLV) (1, 2). This immune response appears to be strain specific, however, since mice of high leukemic strains and mice of some low leukemic strains do not produce appreciable amounts of antibody against MuLV. The failure of mice of high leukemic strains to produce antibody against MuLV is not the result of immunologic tolerance induced by persistent viral replication, since F1 hybrids of mice of high and low leukemia strains produce high titers of antiviral antibody (3, 4). Recently, we have described that the immune nonresponsiveness of AKR mice is genetically controlled by an immune response (Ir) gene in the IXth linkage group (3). We describe here a similar observation with mice of the high leukemic PL strain, thus suggesting that a genetically controlled immune nonresponsiveness to MuLV may be a general characteristic of mice of high leukemic strains.
When Is a Mouse “Old”?
Authors: Walford Roy L.
Published: December 2022

Sir: In recent years the discipline of gerontology—at one time a domain occupied almost entirely by physiologists approaching retirement—has achieved sufficient maturity that investigators experienced primarily in other areas have begun to pursue investigations in the realm of “aging” research. By “aging” I refer to late-life changes, although in another context the term might equally apply to any temporal alterations in function regardless of where they fall in the animals' lifespan (i.e., to a special case of developmental biology).

There are two reasons for an awakened interest in gerontology on the part of immunologists: 1) analogous to the neonatal period, the period of advanced age tends to emphasize or dissect certain features of the immune response (suppressor cell activity, for example), and may present opportunities for fundamental studies in immunology itself; 2) the immune functional changes which occur during aging may contribute substantially to the pathogenesis of those diseases which show a peak incidence in late life, including cancer, vascular disease, maturity-onset diabetes, and others (1).
Generation of Specific Helper Cells and Suppressor Cells in Vitro for the IgE and IgG Antibody Responses
Authors: Ishizaka Kimishige , Adachi Toshihiro
Published: December 2022

Normal splenic lymphocytes from BDF1 mice were cultured on ovalbumin (OA)-bearing syngeneic peritoneal adherent cells for 5 days and their subsequent helper function was tested by an adoptive transfer technique. Lymphocytes harvested from the culture were mixed with DNP-KLH-primed spleen cells and transferred into irradiated syngeneic mice followed by challenge with DNP-OA. The results showed that the cultured lymphocytes has helper function for both IgE and IgG anti-DNP antibody responses. Depletion of mast cells and T cells in the peritoneal adherent cell preparations did not affect the generation of helper cells in the culture. The helper function of the cultured lymphocytes was abolished by the treatment with anti-ϑ antiserum and complement and was specific for ovalbumin. The OA-specific helper T cells were generated in vitro by the culture of a T cell-rich fraction of normal spleen on T cell-depleted OA-bearing peritoneal cells. If the normal splenic lymphocytes or T cell-rich fraction were cultured with 10 µg/ml of OA in the absence of macrophages, cultured lymphocytes lacked helper function. The transfer of splenic lymphocytes or splenic T cells cultured with soluble OA to normal non-irradiated mice, however, suppressed both IgG and IgE antibody responses of the recipients to subsequent immunization with D-NP-OA. The suppressor cells were sensitive to anti-ϑ antiserum and complement and their activity was specific for OA. The cultured cells transferred into normal mice did not suppress anti-hapten antibody response to DNP-KLH. Normal lymphocytes cultured on OA-bearing macrophages and had helper function in adoptive transfer experiments failed to suppress antibody response of nonirradiated recipients to DNP-OA. The results indicate that OA-bearing macrophages primed T cells and generated helper T cells, whereas the culture of normal lymphocytes with soluble OA in the absence of macrophages generated suppressor T cells.
The Ultrastructure of C1t, a Subcomponent of the First Component of Complement: An E.M. and Ultracentrifuge Study
Authors: Pinteric L. , Assimeh S. N. , Kells D. I. C. , Painter R. H.
Published: December 2022

C1t is a 9.5 S α1 glycoprotein that has been shown to be a fourth subcomponent of the first component of complement. The m.w. of C1t was found to be 233,000 by sedimentation equilibrium in the ultracentrifuge. A subunit m.w. of 23,000 was obtained by sedimentation equilibrium in 5.95 M guanidinium chloride. No change in either m.w. was produced by prior reduction and alkylation. In the electron microscope characteristic pentagonal figures of 85 Å diameter were observed together with rod-like figures which appear to be stacked assemblies of the pentagonal figures. These observations lead us to propose that C1t is a noncovalent, decameric protein with the subunits disposed at the vertices of two regular pentagons joined at one of their faces. A possible relationship between C1t and the P-component of amyloid is discussed.
Identification of a Butanol-Extractable Human Placenta-Specific Antigen with Alkaline Phosphatase Activity
Authors: Chang Chin-Hai , Angellis Dennis
Published: December 2022

n-Butanol extracts of whole-term placenta from different individuals were prepared, and used as immunogens to raise heterologous hyperimmune sera in rabbits. Upon immunoelectrophoresis the anti-placenta antisera could recognize at least six antigenic components in the placental extract even after they had been completely absorbed with pooled male serum proteins. However, the antisera so absorbed, designated (-PMS) antisera, could still react strongly with several normal adult tissue extracts including kidney. Systematic and quantitative absorptions of the (-PMS) antisera were thus further carried out with individual butanol extracts of normal adult liver, lung, intestine, stomach, kidney, bone, pancreas, spleen, heart, cerebrum, cerebellum, breast, and packed red cells, as well as a composite extract containing equal amounts of each of the 13 adult tissue extracts. Of the six antigenic components in the placental extracts reacting with the (-PMS) antisera the only one which retained its reactivity with the antisera throughout exhaustive absorptions was associated with alkaline phosphatase activity. This immunologic and enzymologic identity was confirmed with homogeneous placental alkaline phosphatase. Extracts from each of three placentae injected into three pairs of rabbits all produced an identical antibody reaction with the unique determinant(s) of placental alkaline phosphatase. The same identity of precipitin reaction was also found with extracts of 14 other placentae against each of these antisera. It thus firmly establishes that placental alkaline phosphatase is a characteristic placenta-specific fetal protein.
Cross Reactivity of Human β2-Microglobulin with Human Granulocyte Colony-Stimulating Activity
Authors: Price G. B. , McCulloch E. A. , Till J. E.
Published: December 2022

Effects of antisera to human β2-microglobulin (β2m) on factors able to stimulate colony formation in culture by human granulopoietic progenitor cells were investigated. The colony-stimulating activity (CSA) present in media conditioned by cultures of human peripheral leukocytes was suppressed by treatment with anti-β2m. This inhibition was not due to a direct effect on the granulopoietic progenitor cells; controls to test for cytotoxicity and for noncytotoxic inhibition of the progenitor cells by anti-β2m yielded negative results. These experiments provide evidence for a relationship between human CSA and β-microglobulin, and suggest a possible analogy between molceules involved in the in vitro regulation of granulopoiesis and products of the major histocompatibility gene complex.
Two Functionally Distinct Anti-Tumor Effector Cells Isolated from Primary Murine Sarcoma Virus-Induced Tumors
Authors: Holden Howard T. , Haskill J. Stephen , Kirchner Holger , Herberman Ronald B.
Published: December 2022

The ability of cells from primary MSV-induced tumors to function as effector cells in vitro was evaluated. Host cells were isolated by enzymatic disaggregation of the tumor and fractionated by sedimentation velocity at unit gravity on a Ficoll gradient. Characterization of these cells indicated that 30 to 40 % were T lymphocytes, about 50% were macrophages and less than 5% were B lymphocytes. Two different functional activities were mediated by these cells: cytolysis, as measured by the CRA, and inhibition of proliferation, as measured by the GIA. The effector cells in the CRA were T cells with sedimentation velocities of 3.5 to 4.0 mm/hr, whereas those cells which mediated the GIA were presumably macrophages and displayed a heterogeneity in size with two peak sedimentation velocities, one at 4.0 mm/hr and another at 6.0 mm/hr. Activity by the effector cells in the CRA was antigen specific in contrast to the activity in the GIA which was directed against cells which did not carry detectable cross-reacting antigens.
Complement-Mediated Release of Histamine from Human Basophils
Authors: Grant J. Andrew , Settle Linda , Whorton Elbert B. , Dupree Elton
Published: December 2022

Release of histamine from human basophils was induced by activation of complement using zymosan. The histamine-releasing factor resembled C5a on the basis of m.w. (15,000) as well as previous studies showing inactivation by anti-C5. Complement-induced release of histamine was compared with allergic release of histamine which is mediated through appropriate allergens and reaginic IgE. Previously we demonstrated that the former reaction occurred more quickly. Both reactions were inhibited by drugs which increase intracellular concentrations of cAMP3 (theophylline, prostaglandin E1, and histamine) or which mimic the action of cAMP (its dibutyryl derivative). Calcium was required for complement-mediated release of histamine and an increasing response was observed up to physiologic concentrations (2 mM). Magnesium (0 to 1 mM) did not affect the amount of histamine released. Also, glycolysis was probably required for optimal release by complement, since both 2-deoxyglucose and iodoacetamide were inhibitory. When basophils were partly enriched by depletion of neutrophils and eosinophils, the percentage of histamine released by complement was unchanged. Finally, it was shown that activated complement desensitized basophils from responding to a second challenge by the same stimulus. Cross-desensitization was not observed between complement and pollen allergens.
Structure and Biologic Functions of Human IgD
Authors: Kermani-Arab Vali , Burger Denis R. , Leslie Gerrie A.
Published: December 2022

Human peripheral lymphocytes with the capacity to be stimulated by anti-δ exhibited in PHA responsiveness when cultured with anti-δ 1 or 12 hr before PHA exposure over cells exposed to PHA alone. When these lymphocytes were preincubated with PHA 1 or 12 hr before anti-δ activation, no augmentation of the PHA response was seen. In addition, lymphocytes from donors with a high PHA response (low anti-δ activation) failed to show an enhancing effect on PHA responsiveness when pretreated with anti-δ. Moreover, anti-µ showed no synergistic effect on PHA responsiveness. This study is the first to indicate that anti-δ-activated cells enhance PHA responsiveness.
Cold Activation of Complement
Authors: Kondo Motoharu , Hosokawa Keimei , Masuda Masasuke
Published: December 2022

Determination of the complement titer in the serum and plasm of 120 patients with chronic liver diseases showed that in eight (7%) patients with cirrhosis of the liver, chronic active or chronic inactive hepatitis complement in the serum was less than half in the plasma. The dissociation of complement serum and plasma was due to cold activation of the classical pathway of complement in vitro since serum drawn from these patients at 37°C lost hemolytic activity in 4 hours when transferred to a cold environment. Neither HB antigen nor cryoglobulin participated in this phenomenon. The activation of complement in the cold could be prevented by increasing the ionic strength, or by adding vitamin E or, to a lesser extent its vehicle HCO-60, while heparin, Trasylol, soybean trypsin inhibitor, or hirudin had no effect. Trans-AMCHA prevented activation in one case. It is speculated that a factor appearing as a result of blood clotting is able to activate the classical pathway of complement in the cold; it is probably not related to Hageman factor (factor XII), factor VII, thrombin, kallikrein.
Neonatal Infection with Mouse Thymic Virus
Authors: Cross Sue S. , Morse Herbert C. , Asofsky Richard
Published: December 2022

Neonatal infection with mouse thymic virus (TA), a murine herpes virus, produced extensive but temporary necrosis of the thymus which was maximal at 10 to 14 days of age. Studies of precursor and amplifier cells mediating graft-vs-host (GVH) reactivity of thymocytes, spleen cells (SC), and lymph node cells (LNC) of normal and TA-infected mice were made at 4 and 8 weeks of age. Infection with TA resulting in a profound reduction (70 to 80%) in the direct GVH reactivity of thymocytes at both ages; by comparison, the capacity of thymocytes to produce synergy when combined with normal LNC was normal at 8 weeks. Direct GVH reactivity of SC was depressed 90% 4 weeks after infection with TA but returned to near normal at 8 weeks. Direct GVH reactivity of LNC from TA-infected mice was normal at 4 and 8 weeks of age, but amplifier T cell activity in LNC was markedly depressed at 8 weeks. These results demonstrate that TA has highly selective effects upon subpopulations of T cells in thymus and lymph node.
Suppression of Allogeneic Lymphomas in Spleens of Irradiated Mice: Importance of the D End of the H-2 Complex
Authors: Bonmassar Enzo , Cudkowicz Gustavo
Published: December 2022

Dispersed lymphoma cells were transplanted into heavily irradiated mice of a series of congenic resistant strains. Newly synthesized deoxyribonucleic acid was subsequently measured to estimate tumor cell proliferation in spleen and liver. Despite host irradiation, growth of lymphoma cells was suppressed 4 to 7 days after transplantation in the spleen of mice of certain allogeneic strains, but it was barely affected in the liver. The genetic differences resulting in “localized” resistance were mapped in the D region of the H-2 complex. Differences at other regions, including those most important for transplantation resistance in nonirradiated mice (K, I), were inconsequential for lymphoma growth in irradiated hosts. There was a striking resemblance with hemopoietic histoincompatibility, i.e., the rejection of foreign bone marrow grafts by irradiated mice, suggesting that the products of strong Hh gene(s) were expressed in the lymphoma cells.
Early Steps in Specific Tumor Cell Lysis by Sensitized Mouse T Lymphocytes
Authors: Martz Eric
Published: December 2022

In a previous study of the mechanism of specific target cell lysis by alloimmune cytolytic T lymphocytes (CTL), we established that the target cell becomes irreversibly programmed to lyse within a few minutes after contact with the CTL. We here show that at each point in time, the level of specific release of the potassium analog, 86Rb, equals the percentage of target cells which have been programmed to lyse. It is also shown that specific release of 86Rb is more rapid than that of a small metabolite of similar weight, 14C-nicotinamide, which in turn is specifically released more rapidly than 51Cr. Thus, an electrolyte-permeable lesion is produced in the target cell membrane within minutes of contact with the CTL. Since measurements of 86Rb release, unlike measurements of programming for lysis, do not involve exposure of the cells to EDTA and vigorous shearing forces, the present observations corroborate and extend, by an independent and gentler method, our previous conclusion that the CTL effects crucial and irreversible changes in the target cell within minutes after contact. The present results are consistent with the possibility that the first, and perhaps the only damage administered directly by the CTL is a membrane lesion permeable to electrolytes and possible to small molecules.
The Kinetics of Interferon Production by Mouse Lymphocytes and Its Modulating Effect on the Virus Plaque-Forming Cell Assay As a Quantitative Method to Determine Activated Lymphocytes
Authors: Heine Jochen W. , Adler William H.
Published: December 2022

The validity of the virus plaque-forming cell (V-PFC) assay as a means to quantitate stimulated mouse T cells is dependent on the rate and amount of coinciding interferon production which varies with different strains of mice.

In addition, the V-PFC assay demonstrates that the kinetics of cellular activation by mitogens initially involves about 0.01% of responding cells and possibly recruitment of other cells which include cells requiring a longer lag period for activation.
Rapid Detection of Mycoplasma Antibody
Authors: Goldschmidt Barbara L. , Menonna Joseph P. , Dowling Peter C. , Cook Stuart D.
Published: December 2022

Mycoplasma pneumoniae is a significant nonviral cause of acute respiratory disease in children and young adults. The clinical expression of mycoplasmal respiratory tract infection ranges from minor upper respiratory symptoms to acute bronchitis and atypical pneumonia. Indistinguishable respiratory syndromes are commonly caused by several viral agents (1). Mycoplasma pneumonia responds to antibiotics in contrast to the other common respiratory pathogens, but treatment is often inappropriate because mycoplasma infections cannot be distinguished from other causal agents without laboratory aids. Laboratory confirmation by culture is of limited diagnostic use because the organism grows very slowly and may be found in the absence of clinical infection (2, 3). Thus, serodiagnosis, demonstrating changing levels of complement-fixing (CF)3 antibodies to mycoplasma, has become the most widely accepted routine diagnostic tool (4). The CF test, although fairly sensitive and reliable, is a tedious, cumbersome technique, especially for the small laboratory.
The Role of Circulating Interferon in the Modifications of Immune Responsiveness by Mouse Hepatitis Virus (MHV-3)
Authors: Virelizier Jean-Louis , Virelizier Anne-Marie , Allison Anthony C.
Published: December 2022

MHV-3 modifies the humoral immune response to SRBC. During acute infections timing was critical: infecting mice before antigen administration led to immunodepression. Simultaneous injection with virus and SRBC resulted in immunostimulation. Persistent MHV-3 infections were associated with a chronic immunodepression. The presence of circulating interferon (IF) was well correlated with these modifications. IF peaking before antigen was associated with immunodepression whereas IF secretion after antigen was associated with immunostimulation. Low, permanent levels of IF were associated with chronic immunodepression. Since IF is, up to now, the only product of activated lymphocytes that has been shown to modulate immune responses, our results suggest that induction of IF by MHV-3 may be the main mechanism by which this virus modifies immune responsiveness. Moreover, we have shown that MHV-3 infection in susceptible mice diminishes the secretion of lymphocyte IF in response to Sendai virus. In these animals, the thymus cortex was profoundly depleted although the thymus medulla remained unchanged. The MHV-3 infection may, therefore, interfere with a subpopulation of IF-secreting lymphocytes. The possible physiologic role of such lymphocyte subpopulation in terms of host-virus relationships is discussed.
In Vitro Induction of F1 Hybrid Anti-Parent Cell-Mediated Cytotoxicity
Authors: Shearer Gene M. , Garbarino Carol A. , Cudkowicz Gustavo
Published: December 2022

Spleen cells from normal (C57BL/6 × DBA/2)F1 mice were sensitized in vitro for 5 days with irradiated C57BL/6 or DBA/2 parental stimulating cells. Effector cells were generated which specifically lysed 51Cr-labeled targets (leukemia or mitogen-stimulated lymphoid cells) H-2-matched with the parental genotype used for sensitization. The response of F1 spleen cells to the C57BL/6 parent was stronger and more reproducible than that to the DBA/2 parent. The kinetics of generation of effector cells were similar for the F1 anti-parent and an F1 anti-allogeneic response. However, the magnitude of the F1 anti-C57BL/6 cytotoxic response was considerably lower than the F1 response to allogeneic cells. The ratio of responder to stimulator cells in the cultures was more critical for the former than for the latter response. Several lots of fetal bovine serum were found to be adequate for supplementing the medium in the induction of F1 hybrid anti-parent and anti-allogeneic cytotoxic effector cells.

Based on these and other studies, it would appear that the F1 hybrid anti-parent cytotoxic response provides an in vitro model of murine hemopoietic graft rejection in vivo. This response may be elicited by a mechanism distinct from T cell-mediated cytotoxicity and involve different subpopulations of spleen cells.
Immune Response of Mice to the Thy-1.1 Antigen: Intra-H-2 Mapping of the Complementary Ir-Thy-1 Loci
Authors: Zaleski Marek B. , Klein Jan
Published: December 2022

The primary immune response to the Thy-1.1 antigen was measured by a plaque assay that detected cells producing antibodies lytic for AKR thymocytes. F1 hybrids which were b/d heterozygous at the entire H-2 complex or at only some of its regions were studied. All F1 hybrids which were b/d heterozygotes at the K, IA and possibly IB regions displayed a complementary effect of the alleles at the Ir-Thy-1A and Ir-Thy-1B loci. The complementary effect was indicated by the significantly higher responses of the F1 hybrids than the responses of their parental strains. No complementary effect could be demonstrated in F1 hybrids which were b/d heterozygous at the IB, IC, S and D regions, but not at the K and IA regions. The results excluded participation of the IC, S and D regions in the phenomenon of the complementation. Furthermore, one of the Ir-Thy-1 loci could be mapped to the left of the IB subregion, most likely in the IA subregion of the H-2 complex. The second Ir-Thy-1 locus could be mapped to the left of the IC subregion in either the IA or IB subregion.
Participation of Cyclophosphamide-Sensitive T Cells in Graft-vs-Host Reactions
Authors: Schwartz Anthony , Orbach-Arbouys Simone , Gershon Richard K.
Published: December 2022

Cyclophosphamide (CY)3 is toxic for a population of mouse T cells which is active in graft vs host (GVH) responses. This observation contrasts with previous reports which indicate that treatment of mice with CY regimens, similar to those used in this study, enhances the reactivity of T cells which mediate delayed type hypersensitivity to sheep red blood cells. The GVH-inducing capacity of T cells is not altered by removal of B cells by nylon wool columns. We suggest that CY-sensitive T cells may be “regulatory cells,” and that the effect they have depends on, among other things, the nature of the stimulating antigen to which the cells are responding and/or the assay used for study.
Immunosuppression by Spleen Cells from Moloney Leukemia
Authors: Stiller Ronald A. , Cerny Jan
Published: December 2022

The mechanism by which spleen cells from mice bearing established Moloney leukemia suppress the in vitro anti-SRBC PFC response and the mitogen-induced proliferative response was investigated. Suppression of the PFC response was clearly evident when leukemic spleen cells (SMuLV) were mixed with normal responder spleen cells (SN) at a ratio of 1:10. The suppressor cells were most effective when added to the responders during the first 24 to 48 hr of culture. Similarly, suppression was seen when responders were treated with homogenate fractions prepared from SMuLV. No suppression of the PFC response was demonstrated when responder cells were mixed with supernatants harvested from cultures of SMuLV. In contrast, the incorporation of 3H-thymidine by mitogen-stimulated B and T cells was inhibited by supernatants collected from cultures of both SN and SMuLV, however, this suppressive effect was abrogated after dialysis of the supernatants. In addition, responder cells were unaffected when separated from suppressors by a 100-nm Millipore filter in double Marbrook tissue culture chambers. The results of this study lend considerable support to the hypothesis that suppression in the Moloney leukemia model system is not due to the effect of extracellular mediators, but rather is the result of direct cellular interactions between the suppressor and responder cells. The exact nature of the suppressor cell has yet to be determined.
An Indirect Radioimmunoassay for Thymus Leukemia (TL) Antigens
Authors: Esmon Naomi L. , Russell Little J.
Published: December 2022

An indirect radioimmunassay for thymus leukemia (TL)2 antigens has been developed and its specificity documented. The assay makes use of anti-TL antibodies produced in congenic mice (A-Tlab) and radioiodinated purified rabbit anti-mouse IgG. Using this assay, differences can be detected in the amounts of antigen expressed on thymocytes of the three known phenotypes (TL.1,2,3; TL.2; TL-) of inbred mouse strains. Significant differences are also detected in comparison of the thymocytes from homozygous TL.1,2,3 mice (A-Tlaa) and heterozygotes from Tlaa and Tlab parents. Optimum conditions for the assay have been established. Its ability to detect antigens on glutaraldehyde-fixed cells and the binding of noncytolytic antibodies on both viable and fixed cells are documented. The assay has also been used to quantitate the changes in TL antigen expression on cells incubated in anti-TL antisera under conditions of antigenic modulation.
Development of Monoclonal IgA and an Apparent IgG in a Patient with Macroglobulinemia: Sharing of Individually Specific Antigenic Determinants among IgM, IgA, and IgG
Authors: Fair Daryl S. , Schaffer Susan , Krueger Robert G.
Published: December 2022

Patient CM, who initially was diagnosed as having macroglobulinemia (IgM, κ) was subsequently found to develop a monoclonal IgA(κ) protein. Rabbit antisera directed against the patient's IgAm and IgM were rendered specific for individual antigenic (ind) determinants. The anti-IgAm and IgM ind sera reacted with both 131I labeled monoclonal proteins in a double antibody radioimmunoassay (RIA). In addition, both monoclonal immunoglobulins inhibited the reaction between labeled immunoglobulin and both ind antisera, and statistical analysis of the data suggested that the shared ind determinants were identical. The IgG fraction of patient CM's serum also contained a component which competed with both monoclonal IgA (CM) and IgM (CM) in the RIA specific for ind determinants. Analysis of serum samples taken over a 2-year period revealed that, in addition to IgM, both the IgA and IgG components possessing the shared ind determinant(s) were present in low concentrations in the earliest sample, although not detected by conventional techniques. The monoclonal IgA and the igG component were found to increase in concentration over this time interval with a concomitant decrease in IgM. The regulation of immunoglobulin expression with respect to the proposed models of gene organization in antibody-producing cells was discussed.
Circadian Rhythmic Plaque-Forming Cell Response of Spleens from Mice Immunized with SRBC
Authors: Fernandes Gabriel , Halberg Franz , Yunis Edmond J. , Good Robert A.
Published: December 2022

Studies under controlled conditions of lighting and temperature revealed clear evidence of circadian periodicity with respect to the number of PFC present in the spleens of BALB/c mice 3 or 4 days after immunization with SRBC. Striking differences in proliferative responses of spleen lymphocytes to PHA or Con A were also observed at two different circadian times. Large proliferative responses occurred at the time when injection of antigen and/or sampling for PFC yielded a low PFC formation (early in the daily dark span) and small proliferative responses occurred at the time when antigen injection and sampling yielded high formation of PFC (early in the daily light span). These findings indicate that care should be taken to control the circadian timing of stimulation and sampling in studies of immune responses, and that rhythmically varying aspects constitute a new dimension of immunologic processes awaiting further analysis in both the circadian and weekly spectral regions.
The Suppressive Effect of Interferon on the Ability of Mouse Spleen Cells Synthesizing IgE to Sensitize Rat Skin for Heterologous Adoptive Cutaneous Anaphylaxis
Authors: Ngan Johnson , Lee Spencer H. S. , Kind Leon S.
Published: December 2022

Mouse spleen cells which are synthesizing IgE can sensitize rat skin for HACA.2 When such cells are treated with mouse interferon, their ability to sensitize rat skin is reduced or completely inhibited. Two hundred PRD50 units of interferon reduced HACA reactions by more than 50%; 600 units gave complete inhibition. Interferon may act by inhibiting IgE synthesis and/or release.
Immunologic Cross-Reactions Between the Specifically Localizing Cells (SLC) Generated in Response to Sheep RBC and the Red Cells of Other Species
Authors: Thursh Donald R. , Chan Keith S.
Published: December 2022

Some of the lymphoid cells selectively incorporating radioactive thymidine 3 days after primary immunization with SRBC which are capable of specifically localizing in the lymph nodes of adoptively immunized syngeneic recipients challenged with SRBC (SLC-SRBC)2 are also capable of specifically localizing in lymph nodes challenged with the red blood cells of a variety of other mammalian species. The one nonmammalian RBC tested, CRBC, failed to cross-react with SRBC by this parameter even qualitatively, although it remains possible that a feeble cross-reaction might be demonstrated with large enough experimental groups. As expected, the magnitude of the observed cross-reactions seems to vary inversely with the phylogenetic distance between the species, with ORBC showing the strongest cross-reaction (41 to 49%), ARBC showing the weakest (8 to 13%), and BRBC and HRBC occupying intermediate positions (13 to 20%). Cross-reactions between secondary anti-SRBC antibodies and the red cells of the other species were weak and inconstant, suggesting that the observed cross-reactions of specific localization may involve T cells more than B cells. During the course of these studies, it was also possible to verify experimentally our impression that with existing methods of selectively labeling SLC it is possible to study the phenomenon of specific localization using only one population of labeled cells, providing certain essential controls are employed.
Coexistence of Helper and Suppressor Activities in Carrier-Primed Spleen Cells
Authors: Chan Eva L. , Henry Claudia
Published: December 2022

Both helper and suppressor activities for an in vitro IgG anti-hapten (Lac) response to Lac-HRBC were demonstrated in the same population of carrier (HRBC)-primed spleen cells. The relative radiosensitivity of suppressor activity permitted the selective removal of suppression and the demonstration of help. Both activities appear to be T cell-dependent and antigen-specific. Dilution analysis showed that help and suppression are mediated by distinct populations of cells.