bluuebunny/crossref_metadata_2025 huggingface dataset:
Non-T Cell Killing of Mammary Tumor Cells by Spleen Cells: Secretion of Antibody and Recruitment of Cells
Authors: Blair Phyllis B. , Lane Mary-Ann
Published: December 2022

Both T cell-mediated killing and non-T cell-mediated killing of target MTV-induced mammary tumor cells can be detected in microcytotoxicity assay tests of spleen cells from mice immunologically responsive to either the histocompatibility antigens or the virus-associated antigens of the target cells. The non-T cell-mediated cytotoxicity is antibody-dependent; otherwise inactive cells (null cells) can be recruited to activity by target cell-specific factors obtained from the supernatant of short-term cultures of sensitized B cells or provided by the introduction of a small number of sensitized B cells to the wells of the assay plate.
Antibody-Dependent Cell-Mediated Cytotoxicity in Humans
Authors: Trinchieri Giorgio , Bauman Peter , de Marchi Mario , Tökés Zoltán
Published: December 2022

Mononuclear cells from human peripheral blood are able to destroy alloantibody-coated target cells of a human lymphoblastoid line. The effector cell is a nonadherent, non-phagocytic, medium sized lymphoid cell. This cell is not detectable in early human fetuses. Its activity is destroyed by x-ray treatment in vitro but is not inhibited by anti-immunoglobulin sera. Its efficiency in antibody-dependent cell-mediated cytotoxicity is rapidly lost in culture and this loss is prevented by stimulation with allogeneic cells.
Mitogenic Effect of Bacterial Peptidoglycans Possessing Adjuvant Activity
Authors: Damais C. , Bona C. , Chedid L. , Fleck J. , Nauciel C. , Martin J. P.
Published: December 2022

Two purified peptidoglycans extracted from E. coli or B. megaterium strongly stimulated the spleen lymphocytes of rabbits and of normal or nude mice. Both preparations can substitute for Mycobacteria in Freund's complete adjuvant. The peptidoglycan extracted from M. lysodeikticus by a similar procedure which lacks adjuvant activity, did not induce blast transformation. However, the monomere of the E. coli peptidoglycan was devoid of mitogenicity although it has also a marked adjuvant activity.
Resistance to Herpes Simplex Type 2 Virus Induced by an Immunopotentiator (Pyran) in Immunosuppressed Mice
Authors: Morahan Page S. , McCord Ronald S.
Published: December 2022

BALB/c female mice depleted of T lymphocytes by adult thymectomy, lethal irradiation, and bone marrow reconstitution (TXB mice) are approximately 10 times more susceptible than normal mice to i.v. or intravaginal infection with herpes simplex virus type 2. Treatment with the immunopotentiator pyran is effective in protecting both normal or TXB mice against lethal infection with the herpes virus. The results suggest a possible role for immunopotentiator-activated macrophages in the increased resistance afforded mice by treatment with pyran.
Induction of Mouse Type-C RNA Virus by Lipopolysaccharide
Authors: Greenberger Joel S. , Phillips S. Michael , Stephenson John R. , Aaronson Stuart A.
Published: December 2022

BALB/c mouse cells contain genetic information for xenotropic and mouse cell-tropic endogenous type-C RNA viruses. Halogenated pyrimidines induce both viruses from BALB/c embryo fibroblasts and spleen cells in tissue culture. Lipopolysaccharide, and to a lesser extent concanavalin-A, are shown to activate specifically xenotropic virus release by BALB/c spleen cells.
Human Lymphocyte Subpopulation: Early and Late Rosettes
Authors: Yu David Tak Yan
Published: December 2022

In the process of forming SRBC rosettes of human lymphocytes, two types were distinguished, the early and late rosettes. The former appeared immediately whereas the latter required further incubation at 4°C. In this study, the two types of cells were separated by gradient centrifugation and studied separately. It was found that the proportion of caps in the late rosettes was much greater than in the early ones. Treatment of the former by trypsin, AET, and neuraminidase decreased this proportion so that it resembled the late one. It was postulated that some of the differences in the early and late RFC lie in the differences in the content of membrane sulphydryl and sialic acid groups and distribution of SRBC receptors.
Secondary Cell-Mediated Cytotoxic Response to Syngeneic Mouse Tumor Challenge
Authors: Holden Howard T. , Kirchner Holger , Herberman Ronald B.
Published: December 2022

When C57BL/6 mice previously immunized with murine sarcoma virus (MSV) were challenged with a Rauscher virus-induced lymphoma, RBL-5, a secondary cell-mediated cytotoxic response could be detected by the 51Cr release cytotoxicity assay. The level and distribution of the secondary cytotoxic response was affected by the route of challenge. Animals injected i.p. demonstrated a high level of cytotoxicity in the peritoneal exudate cells 3 days after challenge and subsequently cytotoxicity was detected in most lymphoid organs, although at lower levels. However, when the animals were challenged intramuscularly in the leg, the response was not detected as rapidly and furthermore cytotoxic lymphocytes were found only in the draining lymph node and not in other lymphoid organs. Treatment of the effector cells with anti-ϑ and complement showed the secondary response to be predominately dependent on T cells. In addition, the cytotoxicity was specific in that cells lacking cross-reacting antigens were not killed by these attacker cells from mice undergoing a secondary response.
Hyperacute Rejection and Enhancement of Mouse Skin Grafts by Antibodies with a Distinct Specificity
Authors: Jansen Jan L. J. , Koene Robert A. P. , v. Kamp Gerard J. , Hagemann Jacq. F. H. M. , Wijdeveld Paul G. A. B.
Published: December 2022

Pure 7S IgG2 was prepared from B6AF1 anti-B10.D2 ascites fluid by affinity chromatography. This preparation was extensively absorbed with B10.D2 red blood cells to remove the antibodies directed against the serologically defined antigens of the major histocompatibility complex. After absorption the serum had retained in vitro cytotoxic activity against 45% of B10.D2 spleen cells. The absorbed and unabsorbed 7S IgG2 antibodies were tested for their capacity to induce enhancement and, in the presence of rabbit complement, hyperacute rejection of B10.D2 skin grafts in B6AF1 recipient mice. The enhancing capacity of 7S IgG2 was completely unimpaired after absorption. However, the absorbed preparation was unable to induce hyperacute rejection of the grafts even if very high doses were administered. The results show that hyperacute rejection of skin grafts in this model is mediated by antibodies directed against the serologically defined antigens (i.e., H-2K antigens) of the H-2 complex. They further support the hypothesis that immunologic enhancement is mediated by antibodies directed against Ir-region-associated antigens.
Induction of Mouse Homocytotropic Antibodies to Timothy Pollen Antigens
Authors: Fairchild Sally S. , Malley Arthur
Published: December 2022

The IgG1 and IgE homocytotropic antibody responses of LAF1 and C3H mice to timothy pollen antigens are defined. Both mouse strains responded to low doses of crude timothy pollen extract (WST) or a major antigen of timothy pollen coupled to a purified fraction of Ascaris suum (Antigen B-Ascaris). Titers in LAF1 mice were greater than those in C3H mice. Regardless of the immunogen, antigen B was the major determinant recognized by the homocytotropic antibodies; PCA titers with WST or antigen B for challenge were equivalent and PCA activity could be inhibited by antigen D, a dialyzable fraction of timothy pollen possessing the antigen B determinant in monovalent form. The possible usefulness of antigen D for in vivo and in vitro studies of specific immune suppression of cellular activity is discussed.
Class Differences in the Antibody Response of HLC and LLC Mice to Sheep Red Blood Cells
Authors: Silver Donald M. , Chai Chen K.
Published: December 2022

The immune response to sheep red blood cells (SRBC) has been studied in two mouse lines selectively bred for a difference in leukocyte levels. Low leukocyte count (LLC) mice responded to multiple injections of 4 × 107 SRBC with a predominant IgM response whereas high leukocyte count (HLC) mice produced IgG almost exclusively, even after a single injection of SRBC. F1 mice responded like the LLC parent and backcross data implied genetic control at multiple loci. No correlation was found between the class of antibody produced in response to SRBC and the leukocyte count in backcross mice. Large numbers of plaque-forming cells (PFC) were detected in the spleens of HLC mice relative to the number detected in LLC spleens after primary and secondary immunizations with SRBC.
Antigenic Selection of IgG Precursor Subpopulations
Authors: Elliott L. H. , Roszman T. L.
Published: December 2022

The temporal increase in the antigen-binding characteristic of the IgG antibody synthesized after primary immunization is thought to result from antigen preferentially selecting those precursors of the antibody-forming cells with higher affinity receptors. We have investigated this regulatory role of antigen on the selection and expression of IgG precursor subpopulations in rabbit lymph node populations undergoing the secondary anti-bovine serum albumin (BSA) response in vitro. The data indicate that although immunologic memory increased between 1 and 4 months after immunization there was not an increase in antigen sensitivity. We also determined the avidity of the IgG antibody secreted by PFC elicited in the cell cultures. Although the avidity of antibody secreted by PFC elicited with low concentrations of antigen was 2-fold greater than when high concentrations of antigen were employed, analyses of the avidity subpopulations of IgG PFC revealed that memory potential was not restricted to high affinity-primed cells. Moreover, populations of PFC that gave differing avidity values were similar in terms of their plaque diameters and IgG secretion rates.
X Chromosome-Linked Defect of CBA/HN Mice in Production of Tumor-Reactive Naturally Occurring IgM Antibodies
Authors: Martin Sue Ellen , Martin W. John
Published: December 2022

Tumor-reactive naturally occurring antibodies (NOA) could be readily detected in sera of many mouse strains including congenitally athymic (nude) and germ free (gf) mice. Mice of the CBA/HN strain, however, were found to possess low or undetectable levels of NOA against a wide range of tumor cell lines. Genetic studies indicated that the defect in production of tumor-reactive NOA in CBA/HN mice was largely determined by the absence of an X chromosome-linked gene and is probably similar to the known X chromosome defect of this mouse strain in their antibody response to thymus-independent antigens. In spite of the low level of tumor-reactive NOA, CBA/HN mice do not have a high incidence of spontaneous tumors. These findings suggest that if tumor reactive NOA are involved in immune surveillance against malignancy they are unlikely to act directly in a quantitative manner in the detection and elimination of autochthonous tumors.
A Surface Membrane Determinant Shared by Subpopulations of Thymocytes and B Lymphocytes
Authors: Stout Robert D. , Yutoku M. , Grossberg A. , Pressman D. , Herzenberg Leonard A.
Published: December 2022

Utilizing a quantitative fluorescence assay with the fluorescence-activated cell sorter (FACS), we have demonstrated that a rabbit antiserum obtained by immunization with cells of a mouse IgM-producing plasma cell tumor (MOPC104E) is reactive with at least two surface determinants, designated Th-B and ML2, on subpopulations of normal murine lymphocytes. The ML2 determinant is restricted to B lymphocytes. The Th-B determinant is shared by splenic B lymphocytes and a large subpopulation of thymocytes, the latter of which express a 3-fold higher density of Th-B on their surface than do the B lymphocytes. Neither Th-B nor ML2 were found on peripheral T cells or on brain, liver, or kidney cells. The available evidence suggesting that Th-B may be a stem cell determinant that is lost upon maturation is discussed.
T Cell Products Activating Stem Cells: Further Studies on the Origin and Action of the Factor(s)
Authors: Cerny Jan , Waner Elizabeth B. , Rubin Arnold S.
Published: December 2022

We studied the capacity of bone marrow hemopoietic stem cells to form colonies (colony forming units, CFU) in the spleens of irradiated recipient mice following incubation of the cells with T cell-derived mediators in short-term culture in vitro. The mediators were a) stem cell-activating factor (SAF) present in crude form in the supernatant from phytohemagglutinin (PHA); stimulated lymphocytes, and b) immunoenhancing factor (IEF) obtained in partially purified form from the supernatant of antigen-stimulated lymphocytes and characterized by its ability to enhance antibody formation in vitro. Both SAF and IEF increased the number of CFU in the cultured bone marrow cell suspension. However, only SAF, and not IEF, significantly stimulated DNA synthesis in the cultured cells. Furthermore, SAF appears to activate CFU in vitro, rather than merely to promote the survival of active CFU stem cells. Experiments with SAF and bone marrow from different strains of mice indicated that the biologic activity was not restricted by K and Ia regions of H2 complex.
Radioresistance of Cells Responsible for Delayed Hypersensitivity Reactions in the Mouse
Authors: Kettman J. , Mathews M. C.
Published: December 2022

The cells responsible for delayed hypersensitivity reactivity in the mouse act in a radioresistant fashion only if such irradiated cells are injected directly into the site of antigen challenge. Intravenous transfer of irradiated, primed spleen cells does not achieve a transfer of sensitivity to show delayed type hypersensitivity reactions. Transfer of sensitivity by the intravenous route can be effected by injection of non-irradiated spleen cells from primed mice.
Separation of Lymphocytes by Specific Adherence to Cellular Monolayers Containing Protein a of Staphylococcus Aureus
Authors: Ghetie Victor , Sjöquist John
Published: December 2022

A technique was developed for the isolation of lymphocytes according to their surface antigenic markers. The method was based on the general reaction of protein A (SpA from S. aureus) with the Fc-part of an IgG antibody. Monolayers of S. aureus or SpA-coated sheep red blood cells (SpA-SRBC) fix antibody-charged cells specifically; non-fixed cells are easily removed by washing. Alternatively, the monolayers can be treated with a cell surface specific antibody prior to addition of non-charged cells. Monolayer-adhered cells were detached by the addition of lysostaphin (bacterial monolayer) or ammonium chloride (SpA-SRBC monolayer). As an example, Ig-bearing cells were isolated from mouse spleen lymphocytes using a specific rabbit anti-mouse Ig serum for charging either the cells or the monolayers.

The recovery of Ig-bearing cells was approximately 84%. The purity of the cells was approximately 83% and the viability 89%. Antibody-SpA complexes on the surface of isolated cells were removed either by trypsin treatment or by cultivation. Ig on the cell surface is restored after 16 hr of cultivation.
Difference in B Cell Mitogen Responsiveness Between Closely Related Strains of Mice
Authors: Rosenstreich David L. , Glode L. Michael
Published: December 2022

Spleen cells from C3H/HeJ mice respond very poorly to the mitogenic action of endotoxin (LPS) in vitro whereas cells from a very closely related strain, C3H/HeN respond very well. An analysis of the genetic similarity of these two strains was performed. There is no significant MLR measurable between cells of these strains under a variety of culture conditions. They do not manifest a significant graft-vs-host response nor do they reject reciprocal skin grafts. Because of this genetic similarity, it is possible to reconstitute the LPS response of nonresponder mice by an adoptive transfer of spleen cells from C3H/HeN mice. This strain pair should therefore prove useful in analyzing the genetic and cellular basis of endotoxin reactivity.
Existence of the VHIII Subgroup in Rabbit Antibody Heavy Chains
Authors: Wang Lom-Seng , Hora James , Friedenson Bernard
Published: December 2022

Sequence analysis of CNBr fragments from the heavy chains of a rabbit anti-p-azobenzoate antibody of restricted heterogeneity has revealed a VHIII-like heavy chain sequence which was present in 6 to 10% of the heavy chain preparation.
Comparative Studies on Monotypic IgMλ and IgGκ from an Individual Patient
Authors: Hopper John E.
Published: December 2022

A comparative idiotypic antigenic analysis of an IgMλ and an IgGκ paraprotein obtained from sera of an individual patient, Br, revealed the presence of very similar idiotypic determinants associated with the VH regions of the Br µ- and γ-chains. In addition, the IgGκ protein expressed light (L) chain-associated idiotypic determinants which were not evident on the IgMλ protein or its isolated L chains.

Extensive tests of specificity revealed that the shared VH idiotypic determinants were not present in large weight amounts of heterologous nonspecific IgM and IgG, nor on Ig molecules contained in a large number of normal and myeloma sera.
Biologic Properties of Transplantation Immune Sera
Authors: Duc H. T. , Kinsky R. G. , Kanellopoulos J. , Voisin G. A.
Published: December 2022

The influence of the course of immunization on the facilitating-enhancing activity of antibody classes has been studied by passive enhancement of growth of A/JAX sarcomas in CBA and IC mice and of C57BL/6 EL 4 leukemia in BALB/c mice. The influence of dilution of antibodies and complexing to antigens was also studied.

During immunization (with several boosters), the enhancing capacity of sera increased together with 7S IgG antibody activity, but showed no correlation with 19S IgM antibody activity. It also was mercaptoethanol resistant. Fractionation and monospecific immunoabsorption showed IgG1 to be more enhancing than an equal number of hemagglutinating units of IgG2a.

When concentrated on a small amount (105) of target sarcoma I cells, complement-fixing IC anti-A antibodies were even inhibitory on Sa I allografted to IC recipients.

Progressive dilutions reversed this situation, IgG1 activity disappearing and IgG2 acquiring enhancing activity. After complexing to corresponding antigens IgG2 also (and immune sera with inhibitory properties) acquired enhancing properties.

These results may provide a basis for understanding the discrepancies between the results of several groups of authors studying the class(es) of enhancing antibodies.
Human Lymphocyte Alloantigen(s) Similar to Murine Ir Region-Associated (Ia) Antigens
Authors: Arbeit Robert D. , Sachs David H. , Amos D. Bernard , Dickler Howard B.
Published: December 2022

An anti-HL-A human alloantiserum contained antibodies specific for B lymphocyte alloantigen(s). These antigens were distinct from HL-A antigens and, unlike the latter, were closely associated with B lymphocyte Fc receptors. By analogy to the mouse, we suggest that these antigens are associated with an Ir gene region linked to the HL-A locus.
The Requirement for Macrophage-Lymphocyte Interaction in T Lymphocyte Proliferation Induced by Generation of Aldehydes on Cell Membranes
Authors: Greineder Dirk K. , Rosenthal Alan S.
Published: December 2022

Guinea pig T lymphocyte proliferation induced by sodium periodate (NaIO4) or neuraminidase-galactose oxidase (NG) occurs when lymphocytes and macrophages are cultured together after treatment of either purified T lymphocytes or macrophages with these agents. Regardless of which cell initially bears the modified surface carbohydrate, lymphocyte proliferation requires the presence of viable homologous macrophages and fails to occur when they are replaced with fibroblasts, erythrocytes, L2C leukemia cells, thymocytes, PMN, line I hepatoma cells, or murine macrophages. Lymphocyte proliferation resulting from NaIO4 or NG treatment of lymphocytes is diminished when these cells are treated with proteolytic enzymes or aged in in vitro culture for 48 hr. By contrast, proteolytic enzyme treatment or in vitro aging has no effect on the ability of NaIO4 or NG-treated macrophages to induce lymphocyte proliferation. The requirement for macrophage-lymphocyte interaction in NaIO4 or NG-induced lymphocyte proliferation is indicative of a central role for the macrophage in the initiation of T lymphocyte proliferation.
Complete Local Tumor Regression with Antibody to Fibrin Fragment E
Authors: Schlager Seymour I. , Dray Sheldon
Published: December 2022

Rabbit antibody to fibrin fragment E (FFE) was used in an immunotherapy model for the treatment of the line-10 ascites variant of a diethylnitrosamine-induced hepatoma in strain 2 guinea pigs. When 0.75 or 1.0 mg of an IgG preparation containing anti-FFE antibody was injected s.c. 6 and 16 days after the injection of a uniformly lethal dose of line-10 tumor cells, complete regression of the i.d. growing tumor was observed in all 18 strain 2 guinea pigs treated. Thus, this therapy appears to be more effective than any BCG or other immunotherapeutic regimen thus far reported for this tumor. No significant anti-tumor effect was noted when normal rabbit IgG or smaller doses (0.25 or 0.50 mg) of the anti-FFE IgG preparation were used. The injection sites exhibited an inflammatory response for 7 to 10 days characterized by erythema and hemorrhage. Since all animals were treated after the metastatic progression of the tumor is known to frequently occur, the long-term tumor-free survival of these animals as well as their resistance to subsequent tumor challenge indicate that the anti-FFE antibody therapy led to systemic tumor immunity.
Studies of Surface Immunoglobulins on Human B Lymphocytes
Authors: Kumagai Katsuo , Abo Toru , Sekizawa Tsuyoshi , Sasaki Mutsuo
Published: December 2022

Lymphocyte preparations isolated from the human peripheral blood were exposed to different acid pH or incubated at 37°C and the presence of immunoglobulin (Ig) on the cell surface was examined by immunofluorescence (IF) tests. Subsequently, such treated cells were incubated in the autologous serum or in the purified IgG, IgA or IgM proteins and their ability to bind each class of Ig was examined. The results showed that IgG molecules dissociated from large proportions of IgG-positive cells upon exposure to pH 4 at 1°C for 1 min or upon incubation at 37°C for 20 min. The cells from which IgG had been dissociated could again combine with IgG, whereupon the number of positive cells increased, being restored to the number equivalent to or higher than those before acid or 37°C treatment. These results indicated that the treatment could elute the cell-bound IgG present on the cell and that the receptor sites were not degraded by the treatment and could combine with IgG. These cell-bound IgG were observed not only on the monocytes, but also on the small lymphocytes. It was also found that certain proportions of mononuclear cells carried the cell-bound IgA that could be dissociated with acid pH or 37°C. No cell-bound IgM was observed on any mononuclear cells. Microscopic observations before and after acid or 37°C treatment revealed that the staining distribution of the cell-bound IgG and IgA on the cell was granular, appearing as a discontinuous fluorescence ring and forming multiple aggregates but no typical polar caps on warming. In contrast, IgG, IgA, and IgM stable to acid or 37°C treatment were found on the lymphocytes but not on the monocytes, and their staining distribution was uniformly diffuse, appearing as a continuous ring and forming a typical cap on warming. Exposure of the cells to pH 4 or to 37°C could also elute the cell-bound IgG passively adsorbed to the human lymphoid cells in a culture, but did not affect the intrinsic S.Ig on the lymphoid cells in a culture or on the lymphoma cells. These results indicate that the exposure of the cells to acid pH or to 37°C may enable us to detect unfailingly S.Ig lymphocytes by removing the cell-bound IgG and IgA present on the monocytes and/or lymphocytes. Thus, an average value of approximately 10% was obtained for the S.Ig lymphocyte in the lymphocyte preparations from 11 healthy individuals. In addition, the results provided the evidence that, even in normal peripheral blood lymphocytes, there may be a population of B lymphocytes which lack the S.Ig but carry the cell-bound Ig.
Peculiar Secretory IgA System Identified in Chickens
Authors: Watanabe Hiroshi , Kobayashi Kunihiko , Isayama Yasuro
Published: December 2022

A homologue of a free secretory component (SC) was identified in chicken intestinal secretion by criteria based on its antigenic relationship with intestinal secretory IgA (SIgA), molecular size, sugar content, and electrophoretic mobility, as well as its elution characteristic from ion-exchange chromatography. SC was obtained in a form free from IgA from the intestinal secretion by salting out and DEAE chromatography, followed by density ultracentrifuguation or Sephadex G-200 gel-filtration. However, the free SC revealed some antigenic deficiency when compared to bound SC of intestinal SIgA and showed a failure of binding to serum-type-polymeric IgA of biliary IgA in vitro.

Several kinds of chicken external secretions were examined for detection of SC and immunoglobulin classes of IgG, IgA, and IgM. In spite of the wide distribution of immunoglobulins in the external secretions, SC antigen could be detected only in intestinal secretion. Most IgA in the secretions had a molecular structure of a tetramer of serum-type IgA, lacking in SC and having 17S to 18.5S and 600,000 to 700,000 daltons. On the other hand, IgA in the intestinal secretion showed close similarity to the mammalian SIgA, associated with SC and having 11.2S and 350,000 daltons. Presence of antibody activity in the intestinal IgA to avian reovirus was confirmed by plaque reduction tests.
Production of Interferon by Immune Lymphocytes Exposed to Herpes Simplex Virus-Antibody Complexes
Authors: Fujibayashi Takashi , Hooks John J. , Notkins Abner L.
Published: December 2022

Splenic leukocytes from rabbits immunized with herpes simplex virus (HSV) incorporated significant amounts of 3H-TdR and produced interferon after in vitro incubation with HSV antigens. In contrast, splenic leukocytes from nonimmune rabbits did not incorporate 3H-TdR or produce interferon when incubated with HSV antigens. Antigenantibody complexes consisting of HSV antigens and anti-HSV antibody also were capable of stimulating immune leukocytes to incorporate 3H-TdR and to produce interferon.
The Xenogeneic Effect
Authors: Farrar John J.
Published: December 2022

Modified Marbrook culture vessels with two chambers separated by a 0.2-µ porosity membrane have been utilized to show that antigen-stimulated human lymphocytes produce a soluble factor(s) which restores the ability of thymectomized, irradiated, and bone marrow-protected mice to mount a primary IgM plaque-forming cell response in vitro. In the initial experiments, the human lymphocytes plus antigen (sheep erythrocytes) were cultured in the lower chambers of the Marbrook vessels and the T celldeficient mouse spleen cells plus sheep erythrocytes were cultured in the upper chambers. The response of the spleen cells was shown to be enhanced as a function of the number of human lymphocytes in the lower chambers. In subsequent experiments, the human lymphocytes were challenged with allogeneic lymphocytes or activated with a variety of T cell mitogens. Supernatants from these cultures, when placed in the lower chambers of the Marbrook vessels, were also capable of reconstituting the antibodyforming cell response of the mouse B cells. The results of the experiments are discussed in relation to a model of B cell induction which incorporates a non-antigen-specific “helper” T cell.
Studies on the Induction and Expression of T Cell-Mediated Immunity
Authors: Kedar Eli , Bonavida Benjamin
Published: December 2022

Two non-overlapping populations of alloimmune cytotoxic T cells with specificity for tumor-associated antigens (TAA) and for histocompatibility antigens (H-2) were characterized by two independent methods. The heterogeneity of cytotoxic cells was demonstrated in spleen cells derived from BALB/c (H-2d) mice sensitized to EL-4 (H-2b) tumor and from C57BL/6 (H-2b) mice sensitized to G-35 (H-2d) tumor cells. Adsorption of immune lymphocytes on monolayers prepared with cells bearing the sensitizing H-2 antigens abrogated the in vitro cell-mediated cytotoxicity (CMC) directed against 51Cr-labeled normal target cells (spleen cells or ConA-activated spleen blasts), whereas significant cytolytic activity to the corresponding 51Cr-tumor cells was still retained. Likewise, in competitive inhibition assays, CMC to 51Cr-tumor target cells was only partially inhibited by unlabeled normal cells, whereas CMC to 51Cr-normal target cells was completely abrogated. These results suggested that alloimmune cytotoxic lymphocytes are heterogeneous and can be subdivided into two independent populations of restricted specificity.

Several experiments suggested that the effector cell population directed against TAA can no longer elicit a graft-vs-host (GVH) reaction in vivo. This was demonstrated by adoptive transfer into lethally-irradiated allogeneic recipients of cytotoxic or primed spleen cells fractionated on host target cell monolayers. Furthermore, these results demonstrated that both effector cells and memory cells possess high affinity binding receptors to corresponding H-2 antigens. The potential use of fractionated immune lymphocytes sensitized to tumor allografts in adoptive immunotherapy is discussed.
The Humoral Response of Mouse Spleen Cells to Two Types of Sheep Erythrocytes
Authors: McCarthy Margaret M. , Dutton Richard W.
Published: December 2022

There is multigene control of the response of mouse spleen cells to two types of sheep erytrocytes (H and L). Discriminator strains of mice make a much higher response to extra antigens found only on H SRBC than to the shared antigens found on both types of erythrocytes. Non-discriminator strains respond only to the shared antigens, making a response as great as the discriminator response to the extra antigens.

The cell type in which this genetic control is expressed was investigated by a number of methods for studying the role of the macrophage, T cell, and B cell in the production of anti-SRBC response.

The results provide no evidence that the gene(s) operate at the macrophage, T cell, or any other accessory cell level. The data from B cell-limiting dilution experiments are compatible with genetic expression at the B cell level.
Studies on the Mechanism of Suppression of Delayed Hypersensitivity by the Antischistosomal Compound Niridazole
Authors: Daniels Jerry C. , Warren Kenneth S. , David John R.
Published: December 2022

Niridazole given in a single oral dose of 100 mg/kg to guinea pigs sensitized to ortho-chlorobenzoyl chloride-bovine γ-globulin (OCB-BGG) regularly abolished delayed cutaneous reactivity. Little effect was observed, however, when cells from these animals were tested in vitro with either direct or indirect assays for migration inhibitory factor (MIF). On the other hand, sera taken from nonsensitized guinea pigs after they had received 100 mg/kg of niridazole markedly diminished antigen-induced inhibition of migration of sensitized peritoneal exudate cells in vitro. The immunosuppressive effects of such sera could not be produced by niridazole itself, thereby suggesting an effect of niridazole metabolites. This suppressive activity was readily removed from the serum by dialysis. The active serum blocked the production of MIF by sensitized lymph node cells but did not affect the action of preformed MIF on macrophages. The effect of this serum was reversible; lymph node cells incubated for 24 hr with active serum, then washed and reincubated with antigen in normal serum, produced normal amounts of MIF. These studies suggest that metabolites of niridazole, but not the parent compound itself, suppress delayed hypersensitivity in guinea pigs and prevent MIF production by lymphocytes without affecting the macrophage response to MIF.
Functional Activities of Rosette Separated Human Peripheral Blood Leukocytes
Authors: Dean Jack H. , Silva John S. , McCoy James L. , Leonard Connie M. , Cannon Grace B. , Herberman Ronald B.
Published: December 2022

Subpopulations of human peripheral blood leukocytes were isolated by rosette formation and tested for functional activity. E -rosette-forming cells (E-RFC) and EAC-RFC were separated from non-rosetting cells by sedimentation on Ficoll-Hypaque gradients. The efficiency of the method and the purity of the resulting subpopulations were high. E-RFC responded to PHA Con A, allogeneic leukocytes, and PPD, with higher levels of proliferative reactivity than the unseparated lymphocytes while E-RFC depleted, EAC-RFC, and null cells showed only low levels of reactivity. Reactivity to PWM and tetanus toxoid was also restricted to the E-RFC subpopulation, but was lower than that of unseparated cells. A staphylococcal antigen preparation triggered lymphoproliferative reactivity in the E-RFC, E-RFC deleted, EAC-RFC, and the null cell subpopulations. 51Cr release lymphocyte cytotoxicity against a human lymphoblast target cell line was found in the E-RFC and null cell fractions but was not observed with the EAC-RFC subpopulation.
Antibody-Dependent Cytolytically Active Human Leukocytes: An Analysis of Inactivation Following in Vitro Interaction with Antibody-Coated Target Cells
Authors: Ziegler H. Kirk , Henney Christopher S.
Published: December 2022

A leukocyte population consisting of approximately 85% lymphocytes, prepared from human peripheral blood by centrifugation through a Ficoll-Hypaque gradient, was studied for its capacity to destroy antibody-coated human liver (Chang) cells in vitro.

Cytolysis was a rapid event: increased ionic flux (86Rb) from the target cell occurred within 10 min of the addition of effector cells. Kinetic analysis of target cell destruction (51Cr release) was compatible with a “one hit” hypothesis, thereby indicating that cytolysis resulted from a single collision with an effector cell. The initial rate of cytolysis was linear and related to the number of leukocytes added, but lysis at all of the leukocyte to target cell ratios tested ceased after 5 hr. The number of target cells killed at that time was directly proportional to the number of leukocytes added.

While the lytic capacity of the effector population was totally depleted after incubation with antibody-coated target cells, cytotoxicity was not affected by co-culturing leukocytes with Chang cells treated with pre-immune serum. The cytotoxic effector cells functioning in this antibody-dependent lytic system are thus to be contrasted with killer T cells, whose lytic activity is not compromised by interaction with homologous target cells.

It was estimated that approximately 4% of the leukocyte population employed could kill antibody-coated Chang cells, a figure consistent with the estimated frequency of “null” cells within human peripheral lymphocytes.
Heterogeneity of Human T Lymphocytes to Bind Sheep Erythrocytes and Mitogenic Responses of Their Subpopulations
Authors: Sasaki Mutsuo , Sekizawa Tsuyoshi , Takahashi Hisashi , Abo Toru , Kumagai Katsuo
Published: December 2022

Heterogeneity of human T lymphocytes was examined by the rosette formation of sheep red blood cells (E-rosette) in two different preaction media: fetal calf serum (FCS) and gelatin Veronal buffer (GVB). Nearly all the T lymphocytes could form rosettes in FCS at 1°C (FCS-R) independently of the incubation time. In contrast, rosette formation in GVB (GVB-R) was apparently dependent upon the incubation time and could not be used in the enumeration of the total T cells even after long incubation. Among the possible subpopulations, a fraction (T1) that could form GVB-R as well as FCS-R, and the other fraction (T2) that could form only FCS-R were partially purified by rosette formation and by density gradient on a Ficoll-Isopaque layer. In contrast to the T cell-depleted fraction (B) which formed EAC rosettes in a majority of cells and E rosettes in a few cells, only a small portion of cells from both the T1 and T2 fractions could form EAC-rosettes. T1 cells possessed high responses to PHA as estimated by the counts of 3H-TdR incorporation as well as the stimulation indices. In contrast, T2 cells possessed a lower responsiveness to the mitogen, and B fraction cells could not respond at all. These results suggested that a positive correlation exists between the ability of E rosette formation and responsiveness to PHA in the T cell subpopulations. Preliminary experiments also suggested that percentages of T1 cells highly responsive to PHA may decrease and, therefore, those of T2 cells less responsive to the mitogen may increase in the T cell population of the patients with some immunodeficiencies.
Lack of Inverse Relationship Between the Net Charge of T-Independent Immunogens and the Responding Spleen Cells
Authors: Falkenberg Frank W. , Karniely Yuval , Schmitt-Verhulst Anne-Marie , Mozes Edna , Sela Michael
Published: December 2022

Previous studies have demonstrated an inverse relationship between the net electrical charge of immunogens and the antibodies they elicit. This correlation was found to be expressed at the cellular level. It has been shown that thymus-derived cells may recognize immunogens on the basis of their overall electrical charge. In this study, charged T-independent copolymers composed of tyrosine, glutamic acid, and lysine of the D configuration were prepared in order to find out whether this net charge phenomenon holds also for immunogens which do not require helper T cells for generation of immune response.

Spleen cells were fractionated over negatively charged glass bead columns, and their immunocompetence was tested by transferring them into irradiated recipient mice which were immunized with the dinitrophenylated acidic or basic T-independent carrier. No differences in the responsiveness to the dinitrophenyl group on either carrier could be detected in recipients of unfractionated or fractionated cells on the charged columns.

Similar results were obtained with the negatively charged T-independent branched polypeptide poly(DTyr,DGlu)-poly(DPro)--poly(DLys). Filtration of spleen cells through glass bead columns did not affect the immune response potential of the recipient mice. In contrast, a significant reduction was observed in the frequency of positive responses in recipients of filtered cells, which were immunized with the negatively charged T-dependent poly(DTyr,DGlu)-poly(DPro)--poly(DLys). Thus, the inverse net charge phenomenon holds only for T-dependent antigens.
Determinants of the Hierarchy of Humoral Immune Responsiveness During Ontogeny
Authors: Sherwin W. K. , Rowlands D. T.
Published: December 2022

A model system of ontogeny was utilized to investigate the development of humoral immunity in both AKR and BALB/c mice. Lethally irradiated adult mice were reconstituted with syngeneic fetal or neonatal liver. These mice were immunized at various times after reconstitution with a series of eight antigens: the bacteriophages F2, øX-174, and T4; the hapten carrier complexes 2,4 dinitrophenyl-bovine serum albumin and fluorescein-bovine serum albumin; and the small proteins: hen egg lysozyme, sperm whale myoglobin, and bovine pancreatic ribonuclease. Subsequent antibody production to the antigens was assayed with either a direct or a modified bacteriophage neutralization technique. Individual mice responded to the various antigens in a sequential pattern which was basically the same for all mice within each strain. However, there was a marked difference between the two strains in the time at which they developed responsiveness to myoglobin.

In order to begin to delineate the separate roles played by B and T cells in the generation of this hierarchical response pattern during ontogeny, the development of anti-DNP and anti-FTC activity was examined in carrier-primed mice. Results of this experiment indicated that functional B cell specificities for the two haptens arise at different times during ontogeny. Further studies are needed to determine whether the hierarchical pattern of immune responsiveness observed for the other antigens is a function of sequential appearance of B cell specificities, T cell specificities, or both.
Ly Antigens: Markers of T Cell Function on Mouse Spleen Cells
Authors: Hirst John A. , Beverley P. C. L. , Kisielow Pawel , Hoffmann Michael K. , Oettgen Herbert F.
Published: December 2022

By the use of Ly antisera and complement it is possible to remove subpopulations of mouse T cells from spleen cell suspension. It is shown that these Ly antisera are specific for the Ly alleles carried by the cells. Helper cells from primed or unprimed mice and cells which respond to Con A express Ly-1 antigen but little or no Ly-2 antigen (Ly-1+ phenotype) whereas cells which respond to PHA express both Ly-1 and Ly-2 antigens (Ly-1+2+ phenotype).
Augmentation of Proliferation and in Vitro Production of Cytotoxic Cells by 2-Me in the Rat
Authors: Soulillou Jean-Paul , Carpenter Charles B. , Lundin Andrew P. , Strom Terry B.
Published: December 2022

Low concentrations (10-5 to 10-8 M) of 2-mercaptoethanol (2-ME) greatly enhance the proliferation of allogeneic cells in the rat mixed lymphocyte culture (MLC). Studies were undertaken to determine the mode of action of 2-ME. MLC proliferation can occur in the absence of serum proteins (fetal calf serum, FCS) only if 2-ME is present; however, a synergistic effect is present with FCS plus 2-ME, with a 3-fold increase in 3HTdR incorporation with FCS concentrations as low as 0.1%. Kinetic studies show no shift in the peak of proliferation (92 hr) when comparing cultures with and without 2-ME; however, 2-ME-supplemented cultures have significant 3HTdR uptake at 24 hr, and the peak amount of uptake at 92 hr is two to four times higher. Delayed addition of 2-ME until 92 and 166 hr produces a further increase in 3HTdR uptake, indicating that the entire effect is not expressed at the time of allogeneic recognition. L-ascorbic acid, another reducing agent which lacks sulfhydryl groups, elicits a much lower effect on DNA synthesis than does 2-ME. The cytotoxicity of cells harvested from MLC supplemented with 2-ME is increased without loss of target specificity, whereas the same concentration of 2-ME has no direct effect upon the cytotoxicity assay except at higher concentrations where 2-ME suppresses cytotoxicity.
Measurement of Absolute Amounts of Antigen-Specific Human IgE by a Radioallergosorbent Test (RAST) Elution Technique
Authors: Schellenberg R. Robert , Adkinson N. Franklin
Published: December 2022

A technique for the absolute quantification of antigen-specific human IgE is described. It employs elution of a calculable amount of antigen-specific IgE from an allergosorbent-antibody complex by means of alkaline pH treatment, followed by measurement of the IgE content of the eluate with a modified radioimmunosorbent test (RIST). With this method IgE antibody directed against the benzylpenicilloyl determinant of penicillin (BPO) was measured quantitatively in sera from seven penicillin allergic patients. IgE specific for ragweed antigen E was measured in sera from 33 ragweed allergic patients. Values obtained for IgE anti-BPO ranged from 19 to 1806 ng/ml and comprised from 1.3 to 27.5% of total serum IgE. Values of IgE anti-antigen E ranged from 9 to 1807 ng/ml, comprising from 3 to 84% of total serum IgE.

Excellent correlation (r = 0.99; p <0.001) was obtained for both antigen systems between values determined by the RAST elution technique and by simple RAST assay with interpolation from a reference serum of known specific IgE content as determined by the elution technique. This suggests that the RAST elution technique may be needed only for primary standardization of reference sera.
Studies on T Cell Clonal Expansion
Authors: Kamat Ramesh , Henney Christopher S.
Published: December 2022

Adult C57BL/6 mice immunized i.p. with an allogeneic mastocytoma cell line (P815 of the DBA/2 strain) develop cytolytically active T cells. Activity peaks at 10 to 11 days and falls rapidly thereafter. We have observed an anomalous in vitro behavior of effector cell populations harvested during this decline. The lytic activity of spleen cells harvested 12 to 18 days after alloantigen was markedly augmented after culture for 24 hr at 37°C. The augmented lytic activity was caused by T cells and specificity exhibited was identical to the pre-culture population. No augmentation of lytic activity occurred when cells were cultured at 4°C or when they were treated with a protein synthetic inhibitor (pactamycin) at concentrations which did not compromise cytotoxic expression by the starting population. Augmentation of cytolysis was independent of DNA synthesis and did not require the presence of added antigen.

When immune lymphoid cell populations were depleted of effector cells by adsorption onto allogeneic monolayers and then cultured, new killer cells differentiated within a 24-hr period. This process was not augmented by the addition of antigen to the nonadherent cell population.

We interpret these demonstrations to show: i) that there is a pre-killer T cell whose differentiation into an effector cell requires de novo protein synthesis but not cell proliferation, and ii) that the in vivo fall in lytic activity during the primary immune response is at least partially caused by suppression of the differentiation of this pre-killer cell.

Furthermore, we conclude that the pre-killer cell is distinct from a memory T cell because: i) its conversion to an effector cell is antigen-independent and ii) because, unlike the memory cell, pre-killers do not bind avidly to allogeneic cell monolayers.
Histocompatibility and T-B Cell Cooperation in Mouse Radiation Chimeras
Authors: Sado Toshihiko , Kamisaku Hitoko
Published: December 2022

Cooperative interaction between histocompatible as well as histoincompatible T and B cells in vivo was studied with B cell-bearing syngeneic radiation chimeras receiving syngeneic, semiallogeneic, or allogeneic A and T cells in various combinations. The results indicated the following: i) Histoincompatible T and B cells normally did not cooperate successfully for generation of antibody-forming cells. ii) Semiallogeneic (C3BF1) T cells cooperated successfully with parent-type (C3H) B cells developed in parent-type syngeneic chimeras (C3H/C3H), whereas parent-type (C3H as well as C57BL) T cells failed to cooperate with F1 (BC3F1) B cells developed in F1 syngeneic chimeras (BC3F1/BC3F1). iii) T cells obtained from C57BL/C3H or C57BL/C3BF1 chimeras, which were most likely donor (C57BL)- derived, cooperated successfully with C3H-derived B cells developed in C3H/C3H chimeras. iv) Evidence was obtained suggesting that stimulation of antibody response by allogeneic effect in the absence of syngeneic or semisyngeneic helper T cells did not take place in this experimental system. v) With the use of congenic resistant strains, it was shown that alloantigens controlled by H-2 loci, in particular by the K-I region of this gene complex, constituted the barrier of cooperative interaction between histoincompatible T and B cells. vi) Cooperation between H-2 compatible, non-syngeneic T and B cells was also disturbed in varying degrees depending on the strain combinations, thus indicating that cell surface antigens controlled by non-H-2 loci also had a significant role for the cooperative interaction between T and B cells. Problems associated with these findings were discussed.