bluuebunny/crossref_metadata_2025 huggingface dataset:
Tumor-Associated Antigens of Rat Moloney Sarcoma Cells
Authors: Leung Kiu , Feldman Joseph D.
Published: December 2022

Rat Moloney sarcoma cells (MST) were pulsed with 35S-l-methionine for 10 and 60 min and lysed by vortexing in 0.5% deoxycholate, 0.5% NP40, 0.02 M Tris, 0.05 M NaCl, pH 7.5, for 30 sec. The lysate was centrifuged at 16,300 × G for 10 min and the supernatant was co-precipitated with Ig fractions of normal BN serum, normal Lewis serum, BN antiserum to Moloney sarcoma cells (BNaMST), BN antiserum to tumor-associated antigens (BNaTAA), BN antiserum to murine leukemia virus (BNaMuLV), BN antiserum to p30 (BNap30), BN antiserum to gp70 (BNagp70), Lewis antiserum to BN (LeaBN), and BN antiserum to BC5 tumor (BNaBC5). With BNaTAA and BNaMST, a cytoplasmic TAA with m.w. 85,000 was detected. In addition, BNaTAA detected three other species of cytoplasmic TAA with m.w. 220,000, 170,000, and 39,000.
Immunologic Suppression After Oral Administration of Antigen
Authors: Mattingly Jerome A. , Waksman Byron H.
Published: December 2022

Rats given 1010 sheep erythrocytes (SRBC) orally were found to contain specific suppressor cells to SRBC in their Peyer's patches (PP) and mesenteric lymph nodes (MLN) after 2 days of feeding. After 4 days of feeding, similar suppressor cells were found in the thymus and spleen, but they were missing in the PP or MLN. These suppressor cells effectively blocked IgM and IgG plaque-forming cell responses to SRBC in Mishell-Dutton cultures and delayed-type-hypersensitivity responses to SRBC when transferred to syngeneic recipients, but they did not affect responses to horse erythrocytes. The orally induced specific suppressor cells appeared to be T2 cells since their activity was eliminated by in vivo treatment of SRBC-fed rats with anti-rat lymphocyte serum but not by adult thymectomy. Because carrageenan partially relieved the suppression observed in culture, the actual suppressive mechanism may also involve a macrophage.
Membrane Proteins of the P388D1 Macrophage Cell Line: Isolation of Membrane Polypeptides That Bind to the Fc Portion of Aggregated IgG
Authors: D'Urso-Coward Maria , Cone Robert E.
Published: December 2022

Lactoperoxidase-catalyzed cell surface radioiodination was employed to radiolabel membrane polypeptides of a murine macrophage cell line P388D1. Optimal conditions for radioiodination of P388D1 cells were determined and were found to differ from conditions used to label lymphoid cells. SDS-polyacrylamide gel electrophoresis of detergent soluble membrane polypeptides revealed that 9 to 10 molecular species from 1 × 105 to 0.15 × 105 daltons were labeled. Radioiodinated, Triton X-100 extracted P388D1 membrane polypeptides were subjected to affinity chromatography on aggregated IgG:Sepharose columns. Elution of the bound polypeptides and analysis by SDS-polyacrylamide gel electrophoresis revealed polypeptides with an apparent molecular size of 8 × 104, which possess binding affinity for the Fc portion of aggregated IgG. The 8 × 104 dalton membrane polypeptides do not readily aggregate, are resistant to degradation, are not composed of disulfide-linked subunits, and do not appear to contain much carbohydrate. Cellular binding characteristics paraleled the binding of soluble receptor for sieved fractions of aggregated IgG suggesting that these polypeptides may be responsible for the in situ binding of aggregated IgG to P388D1 cells.
Types of Cells Involved in Antigen-Stimulated and Spontaneous Rosette Formation with Toxoplasma Gondii
Authors: Noel Masihi K. , Werner H.
Published: December 2022

Antigen-binding cells were identified by using rosette formation of Toxoplasma gondii and defined lymphoid populations under different experimental conditions. Treatment of immunized spleen cell suspensions with anti-Thy 1 serum plus complement inhibited 5 to 29% of the rosette-forming cells (RFC). Higher numbers of thymus-derived lymphocyte-RFC were obtained after incubation at 4°C and by the centrifugation method than by simple incubation at 20°C. RFC were also observed with thymocytes. Combined treatment with anti-Thy 1 serum plus complement and depletion of adherent cells indicated that the major proportion, 46 to 70%, of RFC were B cells. Spleenocytes of nu/nu mice formed similarly high numbers of rosettes. Spontaneous RFC were observed in nonimmunized mice with both spleen and thymus populations. Numbers of rosettes varied considerably depending on the method and the source of cell population used. Removal of adherent cells from spleen suspensions resulted in RFC reduction of 14 to 25% in immunized and 14 to 33% in nonimmunized animals. Pretreatment with anti-mouse immunoglobulin inhibited completely the spleen and spontaneous thymus RFC and partially the thymus RFC in immunized animals.
In Vitro Immune Response of Human Peripheral Lymphocytes
Authors: Ohara Junichi , Kishimoto Tadamitsu , Yamamura Yuichi
Published: December 2022

Stimulation of human peripheral blood lymphocytes (PBL) with pokeweed mitogen (PWM) induced consistent increases of intracellular levels of cyclic AMP and cyclic GMP within 15 min. Increases of cyclic AMP were observed in both B and T lymphocyte populations, but increase of cyclic GMP was observed only in the B lymphocyte population. The addition of anti-µ antibody to B cells abolished PWM-induced increase of cyclic GMP without any effect on cyclic AMP response. Anti-δ antibody did not show any inhibitory or stimulatory effect on PWM-induced increase of cyclic GMP or cyclic AMP. Pretreatment of B cells with anti-µ antibody at 37°C for 1 hr inhibited PWM-induced increase of cyclic GMP, whereas pretreatment with anti-µ antibody at 4°C did not show any inhibitory effect on PWM-induced increase of cyclic GMP. The effect of anti-µ-pretreatment was reversible and pretreated cells were recovered from the inhibitory effect of anti-µ antibody after 36 hr culture.
Regulation of Antibody Response in Different Immunoglobulin Classes
Authors: Kishimoto Tadamitsu , Hirai Yoshikatsu , Suemura Masaki , Nakanishi Kenji , Yamamura Yuichi
Published: December 2022

IgE class-specific suppressor factors (IgE-TsF) released from DNP-Myc-primed T cells showed their suppressor effect in an antigen nonspecific manner, although DNP-specific stimulation was required for their induction. The suppressor activity of IgE-TsF was absorbed with the alloantiserum directed to the subregions between I-A and I-E of the H-2 complex. IgE-TsF from BALB/c mice did not show their suppressor effect on the IgE response of C57BL/6J and vice versa. The suppressor activity of IgE-TsF was removed with a DNP-OA-primed B lymphocyte population including an increased number of IgE-B cells, but not with T cells. Incubation of hapten-primed B cells with IgE-TsF for 3 hr at 37°C selectively abolished the anti-hapten IgE antibody response in the adoptive transfer experiment, indicating that the target cells of IgE-TsF were IgE-B cells.
Regulation of the Immune Response to Tumor Antigen
Authors: Perry Linda L. , Benacerraf Baruj , Greene Mark I.
Published: December 2022

The nature and function of suppressor factor(s) elaborated by suppressor T cells in response to certain chemically induced tumors have been further defined. Thus, suppressor factor(s) specific for the S1509a methylcholanthrene-induced fibrosarcoma have been shown to bear determinants encoded by the I-J subregion of the murine MHC since suppressive activity is removed by passage of the factor through an immunoadsorbent composed of anti-I-Jk coupled to Sepharose. No loss of activity was observed after passage of factor through control columns composed of normal mouse globulin. Furthermore, activity could be recovered from the relevant immunoadsorbent by elution with high salt.

The administration of crude suppressor factor(s) to normal animals for 4 days resulted in the development of a population of suppressor cells that act in a manner analogous to the suppressor cell population used for production of factor. These factor-induced suppressor cells are T cells and exhibit an antigen specificity similar to that displayed by the tumor-induced suppressor cells. Thus, tumor-specific suppressor factor(s) bear I-J determinants and are capable of inducing the appearance of suppressor T cells in the nontumor-bearing host, which may then act in a specific manner to limit host responsiveness to tumor antigen.
The Effect of T Cell Growth Factor on the Generation of Cytolytic T Cells
Authors: Baker Paul E. , Gillis Steven , Ferm Mary M. , Smith Kendall A.
Published: December 2022

The development of cytotoxic effector cells through primary allogeneic mixed tumor-lymphocyte culture (MTLC) was found to be accompanied by the production of T cell growth factor (TCGF). Addition of supplemental TCGF to MTLC resulted in the generation of significantly greater quantities of effector cells, and these effector cells displayed augmented cytotoxic activity. The TCGF-induced effect could not be duplicated by the addition of fresh medium or a mitogenic concentration of concananvalin A. Although TCGF augmented the proliferation of antigen-nonreactive cells, antigen-reactive cells appeared to be preferentially stimulated by TCGF. Finally, it was shown that depletion of TCGF from MTLC resulted in an impairment of proliferation and differentiation of cytotoxic effector cells. These findings demonstrate that soluble factors are involved in the regulation of in vitro cell-mediated immune responses in an analogous manner to similar factors that have been shown to regulate humoral immune responses. Therefore, the forces affecting TCGF production may modulate the amplitude of a T cell-mediated cytolytic response.
Structural Studies on the Murine Ia Alloantigens
Authors: Cook Richard G. , Uhr Jonathan W. , Capra J. Donald , Vitetta Ellen S.
Published: December 2022

Murine splenocytes were radiolabeled with 3H- and 14C-amino acids; the Ia alloantigens encoded by the I-A and I-E/C subregions were isolated by immunoprecipitation and analyzed for structural variation by polyacrylamide gel electrophoresis. The I-A subregion products (k, d, and b haplotypes) are composed of two polypeptides, α and β, with m.w. of 34,000 and 26,000 daltons, respectively. Haplotype-associated differences in m.w. were detected in the I-E/C products of the k, r, p, and d haplotypes. The α and β chains of E/Ck and E/Cr are 34,000 and 28,000 daltons, respectively; E/Cp and E/Cd molecules are composed of 31,000 and 29,000 dalton polypeptides. Thus, there is both subregion (I-A vs I-E/C) and haplotype (E/Ck, E/Cr vs E/Cd, E/Cp) associated variation in the m.w. of the Ia alloantigens. Additionally, the covalent vs noncovalent association of the Ia subunits was examined and it was found that the α and β chains of both I-A and I-E/C are not covalently associated. However, the I-A α and β chains tend to associate through disulfide bonds during detergent lysis; the presence of alkylating agents during cell lysis prevents this association, and only free α and β chains are observed under nonreducing conditions.
Properties of H-2L Locus Products in Allogeneic and H-2 Restricted, Trinitrophenyl-Specific Cytotoxic Responses
Authors: Levy Robert B. , Shearer Gene M. , Hansen Ted H.
Published: December 2022

The role of the recently defined L antigen (a second D region product) in allogeneic and TNP-specific syngeneic primary CML responses has been investigated. The lysis by anti-L specific cytotoxic effector cells was not inhibited when the target cells were pretreated with an antiserum directed against K and D, whereas an antiserum against L completely abrogated this response. Therefore, H-2L products are recognized on the target cells independently of H-2K and H-2D locus products. Both A.SW cells as well as B10 cells were found to respond to Ld alloantigens, in addition to Dd alloantigens when stimulated by cells differing only in the D region. The results of cold target blocking and antiserum inhibition experiments failed to detect cytotoxic cells with specificity for L antigens in association with TNP, under conditions in which TNP-specific effectors to K and D antigens were demonstrable. These findings suggest that there is a more limited involvement of H-2L locus products than the H-2K or H-2D locus products in the induction and specificity of these responses.
5′-Nucleotidase Activity in Subpopulations of Rat Lymphocytes
Authors: Barton Randall W. , Goldschneider Irving
Published: December 2022

Rat lymphocyte populations were assessed for 5′-nucleotidase activity by both biochemical and histochemical methods. Enzyme-specific activity was enriched in lymphocyte plasma membrane fractions and was higher in lymph node and spleen lymphocytes than in thymocytes. Histochemical reactions on sections of spleen and lymph node revealed strong activity in the thymus-dependent regions, periarteriolar lymphoid sheath in spleen, and paracortes in lymph node; whereas the thymus-independent regions, follicles, and germinal centers were negative. In cellular depletion experiments, with three different methods to detect 5′-nucleotidase, it was observed that the depletion of T cells, but not B cells, was accompanied by a loss of enzyme activity and a decrease in the percentage of nucleotidase-positive cells. The results suggest that, among members of the lymphocyte series, high 5′-nucleotidase activity is selectively associated with the plasma membranes of peripheral T cells in the rat.
Modulation of Immune Response by Bacterial Lipopolysaccharide (LPS): Cellular Basis of Stimulatory and Inhibitory Effects of LPS on the in Vitro IgM Antibody Response to a T-Dependent Antigen
Authors: Uchiyama Takehiko , Jacobs Diane M.
Published: December 2022

The role of thymus-derived lymphocytes (T cells) in LPS modulation of T cell-dependent antibody responses has been investigated. We have assessed the effect of LPS on the primary anti-TNP response to TNP-SRBC of cultures of whole spleen cells or T cell-depleted spleen cells that were supplemented with various subpopulations of carrier-primed (SRBC) spleen cells. The TNP-PFC response was enhanced in the presence of irradiated SRBC-primed spleen cells by addition of 0.16 to 20 µg/ml LPS, but inhibition was observed when irradiation of primed cells was omitted. Enhancement but no inhibition occurred when added primed cells were first passed through a nylon wool column. LPS-mediated enhancement was dependent on a T cell in the primed population. These results suggest that LPS modulation of antibody synthesis is dependent on two populations of antigen-specific cells that have opposing effects on B cell responses to a T-dependent antigen: a helper cell that is irradiation resistant, nonadherent to nylon wool, and sensitive to anti-T cell serum, and a suppressor cell that is irradiation sensitive and adherent to nylon wool.
Regulation of the Immune Response to Tumor Antigen
Authors: Greene Mark I. , Perry Linda L.
Published: December 2022

Suppressor T cells arising during the development of certain murine methylcholanthrene-induced fibrosarcomas have previously been shown capable of limiting only those effector responses generated against the homologous tumor. Thus, S1509a-induced suppressor T cells inhibit immune reactivity only to the S1509a tumor in S1509a immune mice and have no effect on the rejection of SAI tumors in SAI-immune animals. In contrast to this is the cross-reactivity of effector cells in this system, whereby animals rendered immune to either the S1509a or SAI sarcoma are equally capable of rejecting a challenge of the opposite tumor. The specificity of suppression has been further defined in the present study, which demonstrates that S1509a-induced suppressor cells can inhibit responsiveness only to the S1509a sarcoma, even in the simultaneous presence of both the S1509a and SAI tumors. Furthermore, the suppressor factor that is obtainable from suppressor T cells demonstrates a similar precise specificity in its ability to limit selectively reactivity only against the inducing tumor, regardless of the simultaneous expression of antigens on other tumors recognized by cross-reactive effector cells. These results suggest that the antigenic determinants recognized by effector and suppressor T cells are different, and may provide a model for further dissection of suppressor cell function in vivo.
Expression of I-A and I-E,C Region-Coded Ia Antigens on Functional B Cell Subpopulations
Authors: Frelinger Jeffrey A. , Hibbler Frederic J. , Hill Sharon W.
Published: December 2022

Ia antigens from specific subregions have been examined on functional B cell populations. Expression of both I-A and I-E,C region antigens was demonstrated on cells required for both lipopolysaccharide mitogenesis and polyclonal activation. Similar I-A and I-E,C subregion expression was found on cells required for response to the T-independent antigen, polyvinylpyrrolidone. TNP-specific IgM and hen egg lysozyme-specific IgG plaque-forming cells also express I-A and I-E,C region antigens. No evidence was found for an Ia- population responsive in the systems tested. Further, no evidence of preferential expression of I-A or I-E,C region antigens was observed in any system examined. Therefore, it appears that B cells express both I-A and I-E,C region-coded Ia antigens.
The Mouse Mutant “Motheaten”
Authors: Sidman Charles L. , Shultz Leonard D. , Unanue Emil R.
Published: December 2022

The developmental status of B and T lymphocytes was examined in the mouse mutant “Motheaten”. At all ages, these mice have reduced levels of B cells but similar levels of T cells, as compared to control littermates. Motheaten mice do not develop the mature forms of B cells that appear in control mice at about 3 weeks of age. Instead, B cells from motheaten mice are characterized by a paucity of surface IgD and complement receptors, defective capping behavior of surface Ig, and a decrease in the low-to-intermediate peak of fluorescence intensities seen with a fluorescence-activated cell sorter. Unlike immature B cells from normal mice, immature B cells from motheaten mice are not inactivated by anti-Ig interactions in vitro. However, this may be due to an enhanced in vitro development of new Ig-positive cells. Finally, in vivo, motheaten mice show a spontaneous differentiation of B lymphocytes to Ig-secreting plasma cells, starting at about 3 weeks of age. Our current hypothesis is that the block in B cell development seen in motheaten mice may be apparent rather than real and instead may reflect an overall enhancement of the B cell developmental pathway. The lack of mature-type B cells in motheaten mice is thought to be due to a nonspecific influence driving them to the Ig-secreting plasma cell stage as soon as they are formed.
µ-Chains from a Nonsecretor B Cell Line Differ from Secreted µ-Chains at the C-Terminal End
Authors: Williams P. Brock , Kubo Ralph T. , Grey Howard M.
Published: December 2022

Peptide analysis and carboxypeptidase-A digestion have been applied to compare the C-terminal primary structure of µ-chains from nonsecreting and secreting human B cell lines. The expected release of C-terminal tyrosine was seen with internally labeled secreted µ-chains; however, only a small amount of tyrosine was released from internally labeled nonsecreted µ-chains. The greater quantity released of other hydrophobic amino acids Phe, Val, and Leu, together with differences in the kinetics of release suggested that tyrosine was not the C-terminal residue. Since these other amino acids are not found near the C-terminal sequence of secreted µ-chains (-Ser-Asp-Thr-Ala-Gly-Thr-Cys-Tyr-COOH), these results indicate a structural difference between these two forms of µ-chains. Further evidence of a structural difference between these two forms of µ-chains was obtained by the analysis of cyanogen bromide peptides. The C-terminal octapeptide from myeloma and secreted internally labeled µ-chains could be isolated by Sephadex G-50 chromatography and high voltage electrophoresis. In contrast, no such peptide could be obtained from nonsecreted internally labeled µ-chains. These results also strongly indicate that nonsecreted µ-chains (i.e., membrane) possess a different C-terminal structure from those that are secreted. These data have led to the proposal of a hydrophobic C-terminal trailer sequence being responsible for the affinity of nonsecreted µ-chain for the plasma membrane.
B Lymphocyte Expression of a Recognition Locus for Human Monocytes and Macrophages
Authors: Guerry Dupont , Dusak Betsy A. , Schreiber Alan D. , Cooper Richard A.
Published: December 2022

Monocyte and macrophage interactions with other cell types are critical to certain immune responses. Such interactions usually require close physical contact between mononuclear phagocyte and target cell. We have explored the recognition (binding) of cultured human proliferative B lymphocytes by monocytes and macrophages. These studies demonstrate that monocytes have the capacity to recognize a variety of B lymphoblast and lymphoblastoid cell lines. This capacity is enhanced by the generation of macrophages in culture but does not extend to the PMN. Binding does not occur at 4°C and depends on the presence of either Ca or Mg ions. Normal peripheral blood lymphocytes, the B cells of chronic lymphocytic leukemia, mitogen-transformed T cells, and three T lymphoblast cell lines are not bound by monocytes. Recognition does not require identity at the histocompatibility locus, proceeding in allotypic as well as autologous situations. The recognition locus of proliferating B cells is trypsin sensitive and regenerable with incubation in the absence of serum. Regeneration of this structure is blocked by the protein synthetic inhibitor cycloheximide. B cell interaction with monocytes is not mediated by immunoglobulin G, by exogenously derived complement (C) or C3b or C3d from any source, by heterologous bovine antigens, by Epstein-Barr virus-determined antigens, or by currently defined, B cell-specific, cell-surface determinants. This locus may provide one means of initiating cell-cell interactions important in the host response to viral induced or neoplastic proliferation of B lymphocytes.
Immune Response to a Mammary Adenocarcinoma
Authors: Huber S. A. , Lucas Z. J.
Published: December 2022

Serum from Fischer rats immunized with 107 cytolytic spleen cells from syngeneic rats bearing the 13762A mammary adenocarcinoma inhibits specific cell-mediated cytotoxicity (CMC). The blocking activity is first detected 5 to 7 days after immunization; peak activity is reached 5 days later. Immunization with spleen cells depleted of T lymphocytes by heterologous anti-T lymphocyte serum and complement abrogates the blocking response. The blocking factor specifically inhibits lysis of the mammary tumor; it does not inhibit lysis of a Maloney sarcoma virus-induced tumor cells (MSVT) or allogeneic fibroblasts by specifically immune spleen cells. Sephacryl S-200 chromatography and immunoadsorption to rabbit anti-rat IgG or anti-rat µ-chain columns indicate that 10 days after immunization, 30% of the blocking activity is IgM and 70% is IgG.

The IgG-blocking activities from animals bearing the 13762A tumor and from animals immunized with lymphocytes from tumor-bearing animals have identical inhibitory specificities. Ascites tumor inocula contained up to 14% host lymphocytes. Depletion of T lymphocytes from the inocula abrogate the early appearance of IgG-blocking activities and prolong animal survival. We interpret these results to suggest that the IgG-BF in sera from 13762A mammary tumor-bearing rats is induced by a small number of immune T cells contaminating the ascites tumor inoculum. It is considered likely, but remains unproven, that anti-receptor antibody-like blocking factors also occur in spontaneously arising tumors.
Adherent Cell Function in Murine T Lymphocyte Antigen Recognition
Authors: Rosenwasser Lanny J. , Rosenthal Alan S.
Published: December 2022

The mechanisms by which adherent cells, presumably of mononuclear phagocytic lineage, influence in vitro antigen-specific activation of murine T lymphocytes was examined. Two distinct functions for macrophages could be discerned. One macrophage function is dependent on a soluble factor produced by cultured adherent cells and is most easily studied with complex multideterminant antigens. This factor is neither antigen-specific nor MHC-restricted in its action in that PEC, regardless of haplotype, produce factor in the absence of antigen. A second function, antigen-specific T cell activation, is seen when antigens of more restricted heterogeneity are used, such as those under the control of Ir genes. This latter activity demands identity or partial identity between the antigen-presenting cell and the primed T cell, thus suggesting an additional specific, genetically restricted function for macrophages in in vitro antigen recognition. Whether these adherent cell functions are mediated by all or distinct subsets of cells was not established.
Regulation of Human T Cell Colonies by an Inducing Activity (TCI) Produced by Normal Human and Malignant Mouse Cells
Authors: Gerassi Esther , Sachs Leo
Published: December 2022

Unfractionated human peripheral blood lymphocytes seeded at 5 × 105 cells per 35-mm Petri dish with phytohemagglutinin (PHA) can be induced to form T cell colonies with a cloning efficiency of 1%. Fractionated cells with T cell markers did not form any colonies at this seeding level. T cells were induced to form colonies either by increasing the number of cells seeded, or by adding conditioned medium (CM) from syngeneic or allogeneic T cells cultured with PHA. This CM induced a cloning efficiency of 10% at a seeding level of 5 × 103 cells per Petri dish. The induction of T cell colonies by CM did not require the presence of phagocytic or B cells. The T cell colony-inducing activity in the CM appears to be a trypsin-sensitive protein(s) and was not substituted by thymopoietin. Incubation of T lymphocytes with this inducing activity for 30 min was sufficient to induce colony formation.

Colony-inducing activity for human T cells was also found in CM from normal human unfractionated lymphocytes incubated with concanavalin A, pokeweed mitogen, or lipopolysaccharide, human peripheral blood cells adherent to a Petri dish, Krebs mouse ascites tumor cells, and MGI+D+ mouse myeloid leukemia cells cultured with PHA. The results indicate that the colony-inducing activity for normal human T cells (TCI) can be found in CM from normal human T lymphocytes, human adherent cells, and some malignant mouse cells.
The Structural Heterogeneity among Degraded γ Heavy Chain Disease Proteins
Authors: Franklin Edward C.
Published: December 2022

The amino terminal sequence of seven newly studied and three previously reported γ Heavy Chain disease proteins show that they begin at several sites ranging from position 215–234 in the hinge region. The presence of more than one species of molecule in several of the sera and the susceptibility of this region to proteolytic cleavage, taken together with the finding of a blocked Heavy Chain disease protein in the serum of one and the cells of another patient support the concept that these molecules are the result of proteolytic cleavage of typical Heavy Chain disease proteins.
Evidence that the “Mouse Mastocytoma” Cell Line (MCT-1) Is of Rat Origin
Authors: Siraganian Reuben P. , Metzger Henry
Published: December 2022

A rat basophilic leukemia arose in a Wistar rat fed B-chlorethylamine and could be transplanted by subcutaneous inoculation into newborn rats (1). Because these cells (RBL-1) have been adapted for tissue culture (2–4) and have receptors for rat and mouse IgE (2–8), they have become very useful for studies of the IgE receptor (reviewed in 9) and for studies on mediators (e.g., ECFA and SRS-A) (10–13). However, these cells could not be triggered for physiologic histamine release via an IgE-mediated mechanism nor by noncytotoxic doses of compound 48/80 or the calcium ionophore A23187 (14).

Five mastocytomas were found among 99 mice of several different strains that had been treated with pristane and Abelson virus (J. G. Pumphrey, M. Potter, R. Risser and R. P. Siraganian, Manuscript in preparation). These tumors have the morphologic characteristics of mast cells, contain granules, and produce histamine and serotonin.
Discordance among Cell-Mediated Cytolytic Mechanisms in Cancer Patients: Importance of the Assay System
Authors: Catalona William J. , Ratliff Timothy L. , McCool Robert E.
Published: December 2022

ADCC and SCMC directed against Change cell targets that are mediated by lymphocytes having properties characteristic of K cells were impaired in cancer patients. In contrast, ADCC directed against CRBC targets that is mediated by both K cells and macrophages was normal in cancer patients, whereas SCMC against CRBC that is mediated primarily by macrophages was increased. Thus, there was a discordance among cytotoxic mechanisms in cancer patients with K cell-mediated cytotoxic function being impaired and macrophage-mediated cytotoxicity being enhanced. Regression analysis suggested that these perturbations of cytotoxic function occurred independently. Cancer patients had an increased proportion of circulating macrophages and a decreased proportion of eosinophils, but these abnormalities did not correlate significantly with ADCC or SCMC.
Idiotypic Analysis of Anti-Gat Antibodies
Authors: Ju Shyr-Te , Dorf Martin E. , Benacerraf Baruj
Published: December 2022

The humoral response to poly-(L-glutamic acid60, L-alanine30, L-tyrosine10), GAT, in mice is further characterized by both idiotype and fine specificity analyses. The common idiotype on murine anti-GAT antibodies (CGAT) was identified in anti-GAT antisera from seven additional strains of mice. These data confirm that the CGAT idiotype can be induced in all inbred strains of mice.

Using a partially inbred strain of mice selected for their ability to respond to poly-(L-glutamic acid50, L-tyrosine50), GT, we demonstrated that the GT copolymer is capable of inducing antibodies that express the CGAT idiotype. In contrast, antisera directed against poly-(L-glutamic acid60, L-alanine40), GA, bind GAT but lack CGAT idiotype. These results indicate that GAT molecules contain determinants either similar or identical to those on GT molecules, which are responsible for the induction of CGAT idiotypic antibodies. We also demonstrated that both GAT responder and nonresponder strains of mice can produce anti-GAT antibodies with similar fine specificity patterns and CGAT idiotype. In addition, we demonstrate that antibodies of the IgM, IgG1 and IgG2 immunoglobulin classes express the CGAT idiotype.
in Vivo Cooperation of Murine Lymphocytes Sharing One Haplotype of the Transplantation Genes: A Requirement for Tolerance of the Nonshared Haplotype
Authors: Marušić Matko , Perkins Eugene H.
Published: December 2022

The ability of mouse thymus-dependent (T) and bone marrow-derived (B) lymphocytes to cooperate in the (humoral) rejection of the rat Yoshida ascites sarcoma (YAS) was investigated. Mice of two hybrid constitutions that had one parent of the same strain and the other of a different strain were used. T and B cells thus shared one haplotype of the transplantation genes, but the other haplotype was different so that allogeneic recognition could take place in both directions. To investigate the influence of allogeneic recognition on T-B cell cooperation, mice were also made mutually tolerant by establishing long-term radiation chimeras. Erythrocytes and spleen cells of the chimera were shown by serologic analysis to be donor-type. When the chimera was to serve as tumor host and B cell source, it was thymectomized before irradiation and reconstitution with Thy 1.2 antisera and complement-treated bone marrow (TIR). The evidence of tolerance was markedly reduced anti-host-type reactivity in short-term in vivo [3H]thymidine uptake studies. Successful cooperation, manifested by YAS rejection in TIR mice given splenic T cells, was seen a) whenever the transferred T cells and the cells of the TIR recipient were syngeneic, and b) when the T cells were nonsyngeneic with the cells in TIR recipients but were obtained from chimeric donors and injected into chimeric recipients. Chimerism of only T cell donors or TIR recipients was not sufficient for YAS rejection to be induced. The data suggest that the allogeneic recognition on either T or B lymphocytes may thwart otherwise successful cell cooperation.
Stimulation by Granulocyte-Macrophage Colony-Stimulating Factor of Leishmania Tropica Killing by Macrophages
Authors: Handman Emanuela , Burgess Antony W.
Published: December 2022

The intracellular protozoan parasite Leishmania tropica was found to survive unharmed and to multiply for several days in normal mouse peritoneal macrophages. In contrast, when infected monolayers were treated with GM-CSF, there was a continuous decrease in the percentage of infected cells, reaching less than 10% on day 4 in culture, compared to about 30% in normal controls. Microscopic observations showed an increased number of dead parasites in GM-CSF treated infected cells. Within 5 hr of incubation with GM-CSF, almost 40% of intracellular parasites showed morphologic damage, compared to less than 10% in untreated cells. Pretreatment of macrophage monolayers with pure GM-CSF before infection led to an increased level of phagocytosis of L. tropica parasites as reflected by the percentage of infected cells and the increased number of parasites in each infected cell. GM-CSF treated cultures showed 73% infected cells containing a mean of five parasites per cell, as compared to controls in which only about 50% of macrophages were infected with only two parasites per cell. The number of dead parasites per cell was 5-fold higher in the GM-CSF treated cultures at 2 hr. After 24 hr the percentage of infected GM-CSF treated cells was less than one-third that in the control cultures.
Synergistic Cytotoxicity
Authors: Muchmore A. V. , Decker J. M. , Blaese R. M.
Published: December 2022

In a previous paper we demonstrated that freshly obtained human plasma contained a heat labile nonantibody factor that induced human mononuclear cells to become nonspecifically cytotoxic toward xenogeneic but not allogeneic RBC targets. We now present evidence that this factor has a loose affinity for human monocytes and human T cells and can arm them to kill xenogeneic RBC targets. Furthermore, proteolytic enzymes markedly enhance this arming effect. This ability to be armed by a heat labile component found in fresh human plasma and the fact that proteolytic enzymes markedly enhance cytotoxicity clearly dissociate this model of nonspecific cytotoxicity from previously reported NK models.
Soluble Helper Factor(s) Participates in the Generation of Cell Mediated Cytotoxicity Directed against Syngeneic Tumor Cells
Authors: Fyfe Derek A. , Finke James H.
Published: December 2022

Data presented here suggest that one mechanism by which cytotoxic T lymphocytes are generated against certain syngeneic tumors may involve collaboration between pre-killer T cells and helper T cells via a soluble helper factor. Furthermore both lymphocyte-defined as well as cytotoxicity-defined antigens expressed on tumor cells may be involved in stimulating the generation of specific cytotoxic T cells. A lymphoma cell line (CH-1) derived from an SJL mouse stimulates both a syngeneic proliferative and cytotoxic response. However, heattreatment of the CH-1 cells, which is known to functionally inactivate lymphocyte-defined antigens, reduced significantly the ability of such cells to stimulate proliferation and the generation of cytotoxic T cells. The addition of supernatant fluids from either mixed lymphocyte cultures or from cultures in which syngeneic lymph node cells were sensitized against CH-1 tumor cells, restored the ability of heat-treated tumor cells to stimulate a full syngeneic cytotoxic response. The degree of T cell specific cytotoxicity that was generated was dependent in part on the concentration of helper factor(s) added to the cultures. Furthermore, supernatant fluids containing helper activity were not able to stimulate responder cell cytotoxicity in the absence of heat-treated CH-1 cells. The helper activity, detected in cultures where SJL lymph node cells were stimulated by CH-1 cells, appears to be similar to that present in supernates from allogeneic MLC since neither are antigen specific nor H-2 restricted in their ability to assist in the development of cytotoxic T cells.
Suppression of Cytotoxic Response to Histoincompatible Cells
Authors: Gorczynski Reginald M. , MacRae S.
Published: December 2022

The kinetics of appearance of the precursors of SuppA cells (capable of inhibiting CTLp → CTL) or SuppB cells (capable of inhibiting (stem cells → CTLp) in neonatal mice, as well as the appearance of SuppA/SuppB cells in mice given neonatal innoculations of semiallogeneic spleen cells has been investigated. The data obtained are consistent with the idea that SuppA cells have a natural role to play in the induction of neonatal tolerance, whereas SuppB cells may be more important for the maintenance of the tolerant state. Unlike the level of SuppB cells, the level of SuppA cells in tolerant mice seems to be modulated by the presence of the tolerizing determinants. Data are provided to show that SuppB cells, once induced in tolerant mice, can adoptively transfer specific allograft unresponsiveness to newborn syngeneic mice in the absence of added tolerizing antigen, whereas SuppA cells are not able to do so. These data fit the notion that SuppB cells may be responsible for the phenotype of clonal deletion.
Mechanisms of Lipopolysaccharide-Initiated Rabbit Platelet Responses
Authors: Morrison David C. , Oades Zenaida G.
Published: December 2022

The mechanism of bacterial lipopolysaccharide- (LPS) initiated, complement- (C) mediated rabbit platelet lysis has been examined. The results of these studies support our previous observations that activation of the alternative C pathway is required for platelet lysis and that preparations of LPS that activate only the classical pathway (e.g., lipid A) do not cause lysis. The temporal relationship of the interaction of the LPS with the platelet before the addition of plasma suggests a time-dependent association of the LPS with the platelet. On the basis of a number of experiments, including inhibition with polymyxin B, treatment of LPS with alkali, and blocking experiments with polysaccharide-free LPS preparations, it is concluded that the lipid A region of the LPS molecule is responsible for attaching the LPS to the platelet. Finally, a comparison of the activity of lipid A-associated protein-LPS complexes with protein-free LPS demonstrated that an equivalent extent of platelet lysis was achieved with one-one hundredth the concentration of the former as that required for protein-free LPS. The data suggest that LAP facilitates attachment of the LPS to the platelet.
Lymphocyte Function in Experimental African Trypanosomiasis
Authors: Wellhausen Sam R. , Mansfield John M.
Published: December 2022

Cellular mechanisms of immunosuppression were studied in C57BL/6 mice chronically infected with the human pathogen Trypanosoma rhodesiense. Mice lost the capacity to mount primary in vitro splenic PFC responses to sheep erythrocytes (SRBC) within 3 days of infection. The addition of normal mouse spleen cells to infected spleen cell cultures failed to restore responsiveness and, in addition, resulted in the acquisition of unresponsiveness by the normal cells. Parasites present in infected spleen cell cultures were not directly responsible for the suppression observed. Immunosuppression was attributable to suppressor cells present in infected spleens, as assessed by the ability of infected spleen cells to suppress normal spleen cell responses to SRBC. The suppressor cell population could be removed on nylon wool or Sephadex G-10 columns, was insensitive to treatment with anti-Thy 1.2 serum and C, and could be enriched for by adherence to tissue culture dishes. We suggest, therefore, that the splenic suppressor cell is a macrophage. The suppressor macrophage may be present in low concentration within a larger nonsuppressive pool of splenic macrophages since suppressor effects were not evident when <104 adherent cells were transferred to normal cells in dose-response studies. There was good correlation between the appearance of immunosuppression and the presence of suppressor cells in that infected mouse spleen cells exhibited suppressor cell capacity at the time when such cultures became unresponsive to SRBC. Suppressor cells were absent, however, from lymphoid organs other than the spleen, and infected lymph node cell cultures were capable of generating a significant primary in vitro PFC response to SRBC. On the basis of our studies, we suggest that the spleen is the major target organ for immunosuppression in T. rhodesiense infected mice, and that suppression is attributable to the emergence of a suppressor macrophage population that is restricted to the spleen.
Phenotypic and Functional Effects of the Motheaten Gene on Murine B and T Lymphocytes
Authors: Davidson Wendy F. , Morse Herbert C. , Sharrow Susan O. , Chused Thomas M.
Published: December 2022

Lymphoid cells from C57BL/6 mice homozygous for the me gene exhibit multiple phenotypic and functional abnormalities from as early as one week of age. In the B cell population these include a reduction in the frequency of detectable surface Ig+ cells, alterations in the level of expression of surface IgM and IgD, and increase in the frequency of large cells, plasma cells and TNP-specific plaque forming cells, production of anti-thymocyte antibody, and a greatly decreased response to LPS. Together these findings provide strong evidence for polyclonal activation of B cells. The high level of expression of xenotropic MuLV gp70 by me/me spleen and lymph node cells provides further evidence for lymphoid cell activation.

In preliminary studies, me/me T cells appeared to be phenotypically and functionally less affected by the me gene. The distribution of Thy 1.2 on the surface of spleen and lymph node T cells varied from low to normal and the mitogenic responses to Con A and PHA were depressed. It remains to be determined what the basic deficiency in me/me mice is and whether it affects primarily B cells or all lymphoid cells.
Turnover and Shedding of Ia Antigens by Murine Spleen Cells in Culture
Authors: Emerson Stephen G. , Cone Robert E.
Published: December 2022

Lactoperoxidase-catalyzed radioiodination was used to examine the metabolic fate of surface Ia antigens on murine spleen cells in culture. Ia antigens, detected predominately on splenic B lymphocytes, were lost from the cultured cells with biphasic kinetics: a 4 to 6 hr rapid phase, t½ = 5 hr followed by slow release through 20 hr, t½ = 30 hr. The rapid loss of Ia antigens observed was abolished by both harsh iodination conditions and nonphysiologic incubation conditions. The rapid decline in Ia activity was shown to be due to shedding of intact Ia antigens from the cell and to a predominant release of IA subregion-coded proteins. Release of Ia antigens from the cell was accomplished by replacement at the cell surface, and thus reflected net membrane Ia turnover. Ia shedding was shown to be extremely temperature dependent, reflecting both a comparatively high activation enthalpy and entropy requirement for turnover.
Production of Antibodies Against Phosphocholine, Phosphatidylcholine, Sphingomyelin, and Lipid A by Injection of Liposomes Containing Lipid A
Authors: Schuster Brian G. , Neidig Martin , Alving Barbara M. , Alving Carl R.
Published: December 2022

Liposomes were investigated as a combined carrier and adjuvant to promote the immunogenicity of lipid A derived from endotoxin. Lipid A-containing liposomes, or lipid A alone, were injected once into rabbits either subcutaneously or i.v. Antiserum activity was measured by complement-dependent damage to appropriate indicator liposomes containing trapped glucose. Lipid A in liposomes produced a greater immune response against lipid A than that obtained by injecting identical amounts of lipid A alone, and the response to the liposome-lipid A combination was equivalent to that obtained by a frequently used alternative method, in which the antigen consisted of acid-treated bacterial cells coated with lipid A.

Antiserum, or purified IgG antibodies, from rabbits immunized with liposomes containing lipid A also reacted with liposomes lacking lipid A. The antibodies were inhibited by phosphocholine, and the activity was adsorbed by phosphatidylcholine, or ceramide phosphocholine (sphingomyelin). The anti-phosphocholine antibodies did not cross-react with lipid A, but their binding to liposomes was inhibited by the presence of anti-lipid A antibodies on the surface of the liposomes. An “anti-liposome” response was not observed when either lipid A alone, acid-treated bacterial cells coated with lipid A, or liposomes lacking lipid A were injected.

We conclude that one injection of liposomes containing lipid A produces an immune response against lipid A that is equivalent to that produced by other methods of immunization. Injection of liposomes containing lipid A also produces an immune response against phosphocholine, phosphatidylcholine, and sphingomyelin.
Cyclical Changes in Susceptibility of A Myeloma Tumor (LPC-1) to Immune Destruction
Authors: Celis Esteban , Hale Arthur H. , Russell John H. , Eisen Herman N.
Published: December 2022

One of six transplantable ascites tumors of BALB/c mice was found to become periodically resistant to cytotoxic T lymphocytes (CTL). About 12 days after LPC-1, a myeloma tumor, was transplanted it became resistant to lysis by allogenic CTL (anti-H-2d) and by CTL directed to trinitrophenyl groups or minor histocompatibility antigens. Susceptibility to lysis by all of these CTL was regained within 2 to 4 days after transfer of the resistant cells to a fresh BALB/c host. These changes were recurrent: in each transplantation cycle the early LPC-1 cells were susceptible and the late cells were resistant to CTL. Analyses with antisera (B10 anti-B10.D2) showed that the serologically recognized products of the H-2d haplotype were reduced about 10-fold on the LPC-1 cells that were resistant to CTL.
Lymphocyte Activation by the Tumor-Promoting Agent 12-O-Tetradecanoylphorbol-13-Acetate (TPA)
Authors: Abb Jochen , Bayliss Gary J. , Deinhardt Friedrich
Published: December 2022

TPA, a highly active tumor-promoting agent, is an effective mitogen for primate peripheral blood lymphocytes. Optimal stimulation of human lymphocytes was obtained 4 days after the addition of TPA at a concentration of 7.5 ng/ml. Lymphocyte fractionation experiments demonstrated that both T and B cells incorporated 3H-thymidine significantly in response to TPA. Lymphocyte blastogenesis was not due to the reactivation of latent herpesviruses by the tumor promoter, since similar responses to TPA were obtained with virus-genome positive or negative cells. Increased levels of DNA synthesis were observed when TPA was added to marmoset, baboon, rhesus monkey, or chimpanzee peripheral blood lymphocytes. Canine peripheral blood lymphocytes and spleen cells from guinea pigs, rats, and mice were not stimulated by TPA. These observations suggest that TPA-induced lymphocyte blastogenesis may be useful for studies of lymphocyte activation and of the molecular mechanisms of action of tumor-promoting phorbol esters.