bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on the Question of Conventional Immunoglobulin on Thymocytes from Primitive Vertebrates
Authors: Yamaga Karen M. , Kubo Ralph T. , Etlinger Howard M.
Published: December 2022

An unexpected cross-reactivity between trout immunoglobulin (Ig) and keyhole limpet hemocyanin (KLH) was observed. Rabbit antisera to KLH were capable of binding to radioiodinated trout Ig and, conversely, antitrout Ig reacted with KLH. The cross-reactive antibodies were not found in preimmune sera and did not arise because of a common contaminant in the two immunizing preparations. The molecular basis of the cross-reactivity was found to reside in the carbohydrate moieties. Isolated glycopeptides from KLH and trout Ig were efficient inhibitors of the croos-reactivity. Furthermore, l-fucose was capable of inhibiting the cross-reactivity, whereas other monosaccharides tested did not. Absorption of anti-KLH with trout Ig and anti-trout Ig with KLH effectively removed the cross-reactive antibodies and only slightly affected the titer to their respective homologous antigens. Antibodies with specificity for l-fucose were isolated from anti-KLH and anti-trout Ig sera by passage over affinity columns and elution with the monosaccharide.
Fc-Receptor Heterogeneity of Human Suppressor T Cells
Authors: Hayward Anthony R. , Layward Lorna , Lydyard Peter M. , Moretta Lorenzo , Dagg Martha , Lawton Alexander R.
Published: December 2022

Concanavalin A (Con A) stimulated the IgM-binding subpopulation of human T cells (TM) to suppress the pokeweed mitogen-induced differentiation of B lymphocytes to plasma cells.

Control TM cells that had not been Con A activated did not suppress. The degree of suppression was related to the number of Con A-TM cells added to the cultures and it was abolished by irradiation of the T lymphocytes either before or after the 24-hr culture period with Con A. Suppression did not require the presence of TG cells, whose suppressor potential has been previously established. These findings indicate that suppressor activity is not confined to the TG subpopulation but may be expressed by TM cells also.
Molecular Heterogeneity of Secretory IgA Anti-DNP Antibodies
Authors: Montgomery Paul C. , Skandera Cheryl A.
Published: December 2022

Evidence is presented that demonstrates that the microscale sucrose isoelectric focusing technique can be used to assess the molecular heterogeneity of secretory IgA anti-DNP antibodies. The data indicate that substitution of the DNP group on the bacterial carrier, pneumococcus, can limit the structural complexity of secretory IgA anti-DNP antibodies. The data support the concept that factors governing secretory IgA antibody heterogeneity are analogous to those influencing the heterogeneity of serum IgG antibodies.
Independent Stimulation of Motility and the Oxidative Metabolic Burst of Human Polymorphonuclear Leukocytes
Authors: Bass David A. , Dechatelet Lawrence R. , Mccall Charles E.
Published: December 2022

It has recently been suggested that a single stimulus to the membrane of the polymorphonuclear neutrophil leukocyte (PMN) produces a sequential, stereotyped response involving motility, degranulation, and the oxidative metabolic burst, and conversely, that the chemotactic response is dependent upon the stimulation of the hexose monophosphate shunt (HMPS). We have used small, synthetic substances, known to cause either increased motility or the metabolic burst, to examine whether these events can be stimulated independently. Phorbol myristate acetate (PMA) is a surface active agent that causes marked stimulation of iodination, superoxide production, chemiluminescence, and the HMPS. Such stimulation by PMA did not alter random or directional motility of PMN in the chemotaxis-under-agarose assay. Also, preincubation of PMN with PMA did not deplete their energy source for chemotaxis as demonstrated by a normal chemotactic response to zymosan activated serum. N-formylmethionyl peptides (f-met-phe, f-met-leu-phe) caused a dose-related stimulation of random and directional motility of PMN, but only a very slight stimulation of the HMPS, protein iodination, superoxide production, or chemiluminescence, and this minimal response occurred at more than 1000 times the concentration needed for stimulation of motility. These results indicate that stimulation of motility in the metabolic burst may involve separate events at the membrane of the PMN and that the events are not necessarily interdependent.
Human Cytotoxic Response in Vitro to Trinitrophenyl-Modified Autologous Cells
Authors: Shaw Stephen , Shearer Gene M.
Published: December 2022

The present study is a survey of classes of self determinants that are recognized in association with the trinitrophenyl hapten (TNP) by human T cell effectors sensitized in vitro with TNP-modified autologous cells. Specificity was investigated by studies of direct lysis and cold target cell competition. Interpretation of the results of direct lysis experiments was facilitated by the use of interaction analysis. Studies of effector populations from six siblings demonstrated that on the average approximately 40% of the cytotoxic activity was specific for polymorphic determinants of which most or all were HLA linked. Studies of effectors from six unrelated donors, selectively matched for HLA-A and -B locus determinants, demonstrated that some of the cytotoxic activity was directed against HLA-A and/or -B locus-associated polymorphic determinants, but that the serologically-defined HLA-A and -B locus antigens were not the only polymorphic determinants recognized. Studies by cold target cell competition demonstrated that some of the cytotoxic activity was directed against TNP in association with relatively nonpolymorphic antigens. Thus, there was evidence for considerable diversity of the classes of self determinants that human T cells can recognize in association with TNP. Furthermore, the possibility of immune response gene control of the cytotoxic response to these determinants was suggested by differences between donors in their response to TNP-modified self determinants.
Measurement of Antigen-Specific IgG Autoantibody Production in Vitro
Authors: Eilat Dan , Reeves J. Patton , Steinberg Alfred D. , Schechter Alan N.
Published: December 2022

This report describes the development of a direct, highly sensitive and reproducible microassay for measuring picogram amounts of IgG antibody produced in spleen cultures of NZB/NZW female mice and specific for a well defined nucleic acid antigen (native ssRNA). The spontaneously synthesized antibodies were extensively purified from the culture supernatants. The isolated IgG anti-RNA antibodies had a high affinity, limited heterogeneity, and were specific for RNA as compared with DNA. Spleen cell cultures produced quantities of anti-RNA antibodies sufficient to account for a large proportion of the circulating anti-RNA antibodies in the whole animal. However, our results provide no evidence for the recently published suggestion (Sawada et al., 1977. J. Immunol. 119:355) that autoreactive lymphocytes are released from normal immunoregulatory control during in vitro culture conditions.
The Expression of Ia Antigens on Immunocompetent Cells in the Guinea Pig
Authors: Yamashita Uki , Shevach Ethan M. , Thomas David W.
Published: December 2022

In the present study we examined the expression of I-region-associated (Ia) antigens by guinea pig T lymphocytes stimulated in vitro with antigen-pulsed macrophages. Treatment of lymph node (LNL) or peritoneal exudate (PEL) T cells taken directly from immune animals with anti-Ia serum and complement (C) dramatically reduced their proliferative response to antigen-pulsed macrophages when determined on the 4th day of culture. In contrast, the response of immune T cells that had been selected by culture for a week with antigen-pulsed macrophages and restimulated in a second culture was not affected by anti-Ia and C treatment. This same result occurred with selected LNL or PEL that were initially treated before the selection culture with either normal serum or anti-Ia serum and C. LNL became resistant to anti-Ia serum and C treatment by 3 days of culture whereas antigen-specific PEL were still sensitive at that time. These results indicate that in an immune animal two antigen-specific T cell subpopulations are generated based on their sensitivity to anti-Ia serum and C treatment, but that only the resistant population is selected by in vitro culture. In addition, we demonstrated that the Ia-negative T cell population can only be activated by histocompatible antigen-pulsed macrophages.
Nonspecific Suppressive and Cytostatic Activities Mediated by Human Colonic Carcinoma Tissue or Cultured Cell Extract
Authors: Remacle-Bonnet Maryse M. , Pommier Gilbert J. , Luc Chantal , Rance Roselyne J. , Depieds Raymond C.
Published: December 2022

We have previously shown that soluble extracts from human colonic carcinoma (SCE) were potent inhibitors of the PHA-induced DNA synthesis of normal allogeneic peripheral lymphocytes. From the present study, SCE appeared to have a broad and nonspecific range of activity. The SCE-suppressive effect was observed whatever the lymphoid organ or the animal species used as a source of stimulated lymphocytes. Moreover, we always obtained a complete inhibition of the lymphoproliferative response to all tested mitogens including PHA, Con A, PWM, and, for animal lymphocytes, LPS. In addition to the suppressive activity on lymphocytes, SCE also inhibited proliferation of cultured human CCL6 embryonic intestine cells and HT29 colonic carcinoma cells, and of cultured rat fibrosarcoma cells.

Soluble extracts from HT29 cells (SCCE) were able to mimic the nonspecific suppressive and cytostatic activities of SCE. The suppressive activity was also found in extracts from hepatic metastases (SHME) of a primary colonic tumor. In contrast, soluble extracts from normal liver or nonmalignant colonic mucosa did not interfere with cell proliferation.

These data suggest that both SCE and SCCE contain molecular components(s) that can inhibit a wide variety of proliferating cells, including stimulated lymphocytes.
Biosynthesis of Immunoglobulin in Human Immunoproliferative Diseases
Authors: Hammerton Kerie , Cooper David A. , Duckett Margaret , Penny Ronald
Published: December 2022

The kinetics of synthesis and secretion of immunoglobulin by bone marrow cells from 32 patients with multiple myeloma (17 IgG, 8 IgA, 7 Bence Jones only) and two patients with benign monoclonal gammopathy have been compared. Optimal conditions have been defined for quantitatively measuring the incorporation of radioactive leucine into intracellular and secreted protein and immunoglobulin.

In cells continuously labeled with [3H]-leucine the rate of protein synthesis in the intracellular lysate was constant over the 24-hr-culture period. The amount of labeled intracellular immunoglobulin increased linearly during the initial 2 to 4 hr and then remained constant, indicating saturation of the intracellular pool of immunoglobulin. Labeled protein and immunoglobulin were secreted linearly after an initial lag phase of 2 to 8 hr. The major part of the secreted protein was identified as the myeloma protein and was proportional to the percentage of myeloma cells in culture; this was particularly the case for IgG and Bence Jones myeloma patients. By linear regression analysis the amount of myeloma protein secreted by the cells was directly proportional to the amount synthesized within the cell and in addition correlated with the synthesis and secretion of total protein. These neoplastic plasma cells synthesize and secrete myeloma protein as a major product and appear to be comparable to normal antibody-producing plasma cells.
In Vitro Activation of Suppressor Cells from Spleens of Mice Treated with Radioactive Strontium
Authors: Merluzzi Vincent J. , Levy Elinor M. , Kumar Vinay , Bennett Michael , Cooperband Sidney R.
Published: December 2022

Mice were treated with two 100-µCi injections of 89Sr to deplete marrow-dependent (M) cells. Mice so treated responded normally to immunization with sheep red blood cells (SRBC) in vivo; moreover, spleen cells from 89Sr-treated mice were able to respond to SRBC after infusion into irradiated recipient mice. However, spleen cells from mice treated with 89Sr did not respond to SRBC in vitro and mixtures of normal spleen cells with the latter were also not able to respond in vitro. The discrepancy between in vivo and in vitro responses was abolished by culturing spleen cells for 24 hr before testing their ability to respond to SRBC in the adoptive transfer in vivo. Pretreatment of spleen cells from 89Sr-treated mice with 1000 R of γ-radiation lessened their suppressive activity. The suppressor cells were detected in spleens of athymic nude mice treated with 89Sr. The suppressive activity, after the 24-hr culture period, was not abolished by irradiation and was active in vivo as well as in vitro. Thus, depletion of M cells by 89Sr results in the appearance within the spleen of thymus-independent suppressor cells, which require a short period of in vitro cultivation before becoming functionally active.
Cell-Mediated Cytotoxicity Against Virus-Infected Target Cells in Humans
Authors: Santoli Daniela , Trinchieri Giorgio , Lief Florence S.
Published: December 2022

In a 51Chromium release assay performed without immune sera, purified lymphocytes from randomly selected healthy donors killed significantly more allogeneic virus-infected than noninfected fibroblasts. An incubation period of 18 to 20 hr was optimal for determining cytotoxicity, but increased cytotoxic activity against infected cells was observed as early as 6 hr from the start of the test. No correlation could be found between donors' antibody titers to any virus tested and the cytotoxic efficiency of their effector cells: virus-infected cells were lysed by both seropositive and seronegative individuals. With several different cell fractionation techniques, the effector cell in this system could not be distinguished from the human natural killer cell, which is a lymphocyte with receptors for the Fc fragment of IgG but with no surface immunoglobulin. When lymphocytes were separated on the basis of ability to form rosettes with neuraminidase or AET (2-aminoethylisothiouronium bromide hydrobromide)-treated sheep erythrocytes, the majority of cytotoxic activity was consistently recovered in the nonrosetting fraction. A portion of it, however, was always present in the rosetting fraction and was, again, mediated by lymphocytes carrying receptors for the Fc fragment of IgG.
Triggering of Cultured Neoplastic Mast Cells by Antibodies to the Receptor for IgE
Authors: Isersky Chaviva , Taurog Joel D. , Poy George , Metzger Henry
Published: December 2022

Cell surface receptors for IgE were isolated from detergent lysates of iodinated, IgE-saturated, rat basophilic leukemia cells by precipitation with anti-IgE antibodies followed by chromatography at acid pH. The isolated material showed a single 125I-band (m.w. ∼58,000) on gel electrophoresis in sodium dodecyl surfate and was used to immunize a rabbit. The resulting antiserum was reacted with lysates of surface iodinated mouse or rat tumor mast cells. Analysis of the precipitates on (10%) gel electrophoresis revealed one major peak comprising >80% of the detectable counts and having an estimated m.w. of ∼58,000. The antiserum reacted with detergent-solubilized and cell-bound receptors in the presence of absence of excess IgE; it also inhibited the binding of 125I-IgE. Cultured mouse mastocytoma cells never exposed to IgE released 3H-serotonin when incubated with F(ab′)2, but not Fab′ fragments of the antiserum, which had been rigorously freed of IgE and anti-IgE. The release was inhibited in the presence of excess IgE, was Ca++ dependent, and equaled 80% of the maximum obtained with IgE and anti-IgE. We conclude that aggregation of the receptors for IgE provides the critical signals for cell activation.
Subclass Restriction of Murine Anti-Carbohydrate Antibodies
Authors: Perlmutter Roger M. , Hansburg Daniel , Briles David E. , Nicolotti Robert A. , David Joseph M.
Published: December 2022

Examination of the subclass distribution of murine antibodies directed against groups A and C streptococcal carbohydrate, α-(1→3) dextran and phosphocholine yields the surprising observation that these carbohydrate antigens stimulate IgG responses largely restricted to the rare IgG3 subclass. This subclass restriction is particularly impressive in light of the low circulating levels of IgG3 in nonimmune mouse serum and the failure of a variety of other antigens including proteins and aromatic haptens to stimulate IgG3 antibody production. Attempts to alter the subclass restriction of antibodies with carbohydrate specificity by immunization with carbohydrate-coupled protein have been unsuccessful and indicate that immunoregulation of subclass expression probably occurs at the level of the antibody forming (B) cell. It is therefore conceivable that VH regions of murine immunoglobulins may be restricted to particular IgG subclasses. A similar type of subclass restriction has been reported in human and rat anti-carbohydrate antibodies. This recruitment of a minor immunoglobulin isotype by carbohydrate antigens in several species further supports the concept of immunoregulation at the level of subclass, and suggests that these and other mammals may share a structurally similar isotype with perhaps a common evolutionary origin.
Mechanism of Allosuppression: Evidence for Direct Suppression of Responding B Cells
Authors: Swain Susan L.
Published: December 2022

The mechanism of allosuppression has been investigated. As previously described, negative allogeneic effects result when T cells recognize MHC-encoded alloantigens on responding lymphocytes, preventing the generation of a secondary anti-PFC response in vitro. Several experiments suggested that this suppression was not due to the generation of cytotoxic effectors. First, effective suppression occurred only when T cells, either unprimed or alloantigen activated, were added during the first 24 hr of the responding culture. Even previously generated cytotoxic effectors were relatively ineffective when added late in the responde. Furthermore, no detectable cytotoxic effectors were found in supporessed cultures.

The major target of suppression was the responding B cell. Only B cells carrying alloantigens (thus recognized by the T cells) were suppressed; bystander B cells were little affected. Thus allosuppression appears to involve the recognition by T cells of alloantigens on responding B cells and direct suppression of some early event in the development of these B cells into PFC.

The responses of primed B cells were found to be preferentially sensitive to the suppressive effects of allo-T cells, whereas the response of unprimed B cells was influenced preferentially by the helper effects of alloantigen-activated T cells. It is possible that the state of differentiation of the B cell may determine the outcome of the interaction with regulatory T cells.
Differential Endotoxin Sensitivity of Lymphocytes and Macrophages from Mice with an X-Linked Deffect in B Cell Maturation
Authors: Rosenstreich David L. , Vogel Stefanie N. , Jacques Allen , Wahl Larry M. , Scher Irwin , Mergenhagen Stephan E.
Published: December 2022

The in vitro sensitivity of B lymphocytes and macrophages derived from (CBA/N × DBA/2N) F1 male mice, which carry an X-linked recessive gene that produces defective B cell maturation, was compared to phenotypically normal F1 female mice. B lymphocytes of F1 males exhibit an abnormal mitogenic response to LPS in serum-free culture conditions, which is partially reversed in the presence of serum. In contrast, both resident and thioglycollate-induced peritoneal macrophages of F1 male mice respond normally to LPS. In response to LPS in vitro, F1 male macrophages produce the monokine, lymphocyte-activating factor (LAF) and release prostaglandins. Furthermore, F1 male macrophages are sensitive to the lethal effects of LPS. Therefore, the defective CBA/N gene appears to be expressed only in B lymphocytes and not in macrophages. Since F1 male mice are normally sensitive to the lethal and adjuvant effects of LPS in vivo, these findings suggest that a mature B lymphocyte population is not required for these effects and support the role of the macrophage in the mediation of LPS-induced lethality and adjuvanticity.
Persistent Activity of Suppressor Cells in T Cell-Mediated Immune Response
Authors: Polak Ladislav , Rinck Claude
Published: December 2022

Tolerance to dinitrochlorobenzene contact sensitivity induced by i.v. injection of dinitrobenzenesulfonic acid in guinea pigs is a long-lasting phenomenon (up to 1 year). The tolerogen, however, was traceable in the circulation only up to 3 months after its application. In spite of that, tolerance was adoptively transferred by parabiosis 6 months after being induced. Moreover, active suppressor cells eliminated by cyclophosphamide treatment are able to regenerate in those adoptively tolerized animals.

These results indicate that the tolerogenic injection stimulates precursors of suppressor cells to generate active suppressor cells and memory cells of suppression. The further formation of active suppressor cells from memory cells seems to be tolerogen independent, but the existence of specific stimulator cells for suppression may be considered. These cells may bind undetectable small amounts of tolerogen.

The recovery of suppression might, however, be also due to recovery of suppressor cells which were temporarily inactivated but not destroyed by cyclophosphamide treatment.
Characterization and Genetic Control of the Immune Response to Synthetic Polypeptide Antigens of Defined Geometry
Authors: Singh Bhagirath , Fraga Ester , Barton Moira A.
Published: December 2022

Both humoral and cell-mediated immune responses to the synthetic helical hapten-carrier conjugate poly-Glu-Tyr-Lys(TNP)-(Glu-Tyr-Ala)5 were found to be linked to the major histocompatibility locus in mice and guinea pigs. The responder mouse strains (H-2d haplotype) showed a primary IgM response with an IgG component appearing after the secondary immunization. The antibody response was accompanied by a positive DTH reaction in responder strains. Nonresponder mice (H-2b or H-2k haplotypes) showed neither IgM nor IgG antibodies and the DTH reaction was negative. Administration of the antigen as a complex with an immunogenic carrier was not effective in inducing a response in nonresponder mice. In guinea pig studies, it was found that strain 2 animals were able to mount an antibody response against the TNP-hapten and a DTH response against the polypeptide backbone. Strain 13 animals gave no anti-TNP antibodies at the lower dose levels and DTH activity was entirely negative for all doses of immunizing antigen. Replacement of the TNP hapten by the arsanilazo dipeptide derivative, BOC-gly-ARA-tyrosine, converted the nonresponder strain 13 guinea pigs into complete responders showing antibody and DTH reactions to both the hapten and the polypeptide backbone.
Antibodies to Guinea Pig Lymphokines
Authors: Sorg Clemens , Geczy Carolyn L.
Published: December 2022

It was shown previously, that an antiserum directed against highly purified fractions of migration inhibitory factor inhibits delayed hypersensitivity reactions in vivo and in vitro. Using radiolabeling techniques we determined that the anti-lymphokine serum reacted primarily with three lymphocyte activation products (m.w. 60,000, 45,000, and 30,000) all of which had a similar isoelectric point of 5.2. The cellular origin of this material was investigated. It was found that activated B cells, B leukemia cells (L2C), and growing fibroblasts produced material of a similar m.w. as analyzed on SDS-PAGE. No cross-reaction was found with radiolabeled products of activated murine and human lymph node cells and of SV 40-infected African green monkey kidney cells. The isoelectric point of the reactive material from B cells, leukemia cells, and fibroblasts was determined at 5.2. In addition to material with pI 5.2, lymph node cells also produced material with pI 3.5 to 4.5, which focused at pH 5.0 to 5.4. After neuraminidase treatment macrophage migration inhibitory activity in fibroblast culture supernatants could be absorbed specifically to insolubilized anti-lymphokine antibody. These findings suggests that lymphoid and nonlymphoid cells are capable of producing molecules whose physicochemical and functional properties appear to be identical.
Inhibition of Migration of Tumor Cells in Vitro by Lymphokine-Containing Supernatants
Authors: Cohen M. C. , Goss A. , Yoshida T. , Cohen S.
Published: December 2022

Lymphokine-containing supernatants derived from seven different human lymphoid cell lines and lymphokine-containing supernatants from concanavalin Astimulated murine lymphocytes were found to be capable of reversibly inhibiting the migration of tumor cells in vitro. The tumor cell lines used in these studies were the P815 mastocytoma, Ehrlich ascites, Walker carcinosarcoma, Hepatoma 129, and Sarcoma 37. Preliminary physicochemical evidence suggests that the mediator, here termed TMIF, is distinct from MIF. In any case, these results suggest the possibility that lymphokines other than lymphotoxin or macrophage-activating factors may play a role in tumor immunity.
Intracellular Control of Human Neutrophil Secretion
Authors: Henson Peter M. , Zanolari Bettina , Schwartzman Nannette A. , Hong Steven R.
Published: December 2022

Human neutrophils released the granule constituents myeloperoxidase and lysozyme, but not the cytoplasmic enzyme lactic dehydrogenase, when pretreated with cytochalasin B and stimulated with purified human C5a. Prior exposure to C5a before the cytochalasin B, however, abrogated the subsequent secretory process. Interaction of neutrophils with C5a was shown to result in a concentration-dependent rapid desensitization that could not be overcome by later addition of cytochalasin B or of cytochalasin B and C5a. The effect was relatively stimulus specific in that neutrophils desensitized in this manner could be induced to release granule enzymes by casein or by complement-coated zymosan particles. Cytochalasin B effects on neutrophils appear to mimic those of surface binding of soluble stimuli such as C5a and immune complexes. It is suggested that desensitization in concert with surface stimulation may represent an important intracellular mechanism for limiting neutrophil secretion.
Structural Studies of the H-2D Products of the Mouse Mutant BALB/c-H-2Ddb and the Parental Strain BALB/cKh-H-2Dd
Authors: Nairn Roderick , Nathenson Stanley G.
Published: December 2022

The tryptic peptide profile characteristics of the H-2D glycoprotein, isolated by immunoprecipitation from the MHC mutant mouse strain BALB/c-H-2Ddb, were compared with those of the H-2D molecule from the parent strain BALB/cKh-H-2Dd. At each stage of purification these molecules exhibited identical biochemical properties and on peptide mapping we observed that the Ddb molecule showed no detectable peptide differences from the Dd molecule of the nonmutant parent. These data thus support the concept that the site of mutation in this mutant strain, although located in the D region of the MHC, is distinct from the gene coding for molecules bearing the H-2.4 private specificity.
Role of Blocking Antibody in Disseminated Gonococcal Infection
Authors: Allen Mccutchan J. , Katzenstein David , Norquist Donna , Chikami Gary , Wunderlich Annette , Braude Abraham I.
Published: December 2022

Gonococci (GC) isolated from patients with disseminated infection (DGI) are uniformly resistant to killing by normal human serum (NHS), but are killed in large numbers by normal rabbit serum (NRS). Since resistance to human serum is necessary for DGI, we examined the mechanism of resistance by GC to NHS.

Our bactericidal assay exposes 10-fold dilutions of 108 plate-grown GC/ml to undiluted serum and measures the number of GC the serum can kill. Killing is expressed as the reduction in GC compared with paired, heat-decomplemented controls.

NRS sterilized >105 GC/ml of six DGI strains. When NRS was mixed (1:1) with nonbactericidal NHS, killing was reduced to <103 GC/ml. This potent inhibitor of rabbit serum could be adsorbed to preincubated and washed GC. The heat (65°C) lability of the inhibitor suggested blocking antibody. Human cord serum containing maternal IgG, but no IgM or IgA, blocked rabbit serum as effectively as normal adult or matched maternal serum. IgG fractions added to rabbit serum reduced killing of DGI strains, but IgM and IgA did not. Absorption of NHS with resistant strains increased killing of the same and other resistant strains and removed the ability to block killing by rabbit serum. Absorption with sensitive strains decreased killing. Thus, resistant strains adsorb blocking antibody and sensitive strains adsorb killing antibody.

We conclude that NHS contains an IgG blocking antibody that competes with natural bactericidal antibody in man and rabbits. We believe that natural IgG binds to certain gonococci possessing the appropriate receptor and enables them to disseminate in the blood by protecting them from killing by antibody and complement.
In Vitro Growth of Murine T Cells
Authors: Rosenberg Steven A. , Spiess Paul J. , Schwarz Susan
Published: December 2022

Supernatants of murine lymphoid cultures stimulated with Concanavalin A contain factor(s) capable of sustaining continuous growth of murine lymphoid cells in vitro. Methods for the optimal generation of these growth factors (GM) have been defined, including number of cells required, optimal concentration of Concanavalin A, incubation time, and strain differences in GM production. With GM, cells have been grown for over 15 10-fold generations over a 2-month period. The growing cells express the θ surface marker and do not express surface Ia antigens.
Studies on the Terminal Stages of Immune Hemolysis
Authors: Boyle Michael D. P. , Langone John J. , Borsos Tibor
Published: December 2022

The inhibitory effects of metal salts on the formation and transformation of E* to ghosts was studied. Ferrous and ferric salts inhibited the binding of C9 to EAC1–8 cells but had no other inhibitory role in the reaction. Uranyl, copper, and zinc salts inhibited the transformation of the EAC1–9inserted intermediate to E ghosts. This inhibition occurred at a stage prior to detectable damage as measured by 86Rb or hemoglobin release and at a step prior to that inhibitable by high molarity EDTA. Consequently a further step in the reaction sequence of E* to ghosts was identified. The reaction sequence leading from EAC1–9 to ghosts can be summarized as follows:
Synthesis of IgM by Cells of NZB and SWR Mice and Their Crosses
Authors: Manny Noga , Datta Syamal K. , Schwartz Robert S.
Published: December 2022

Spontaneous and LPS-induced secretion of IgM by cultured spleen cells of NZB and SWR mice were measured after a 4-hr pulse with 3H-leucine. Similar experiments were done with fetal liver cells of NZB, CBA, BALB/c, and C57BL/6 mice. In short-term (4-hr) cultures there was marked hypersecretion of IgM by NZB cells relative to SWR cells. This abnormality was attributed to two factors: an increased number of IgM-containing cells and an increased secretion of IgM per cell. These two factors segregated as independent phenotypes in the progeny of the crosses. Liver cells from 18- to 19- day NZB fetuses matured into IgM hypersecretors in vitro in the absence of T cells. This phenomenon was uninfluenced by addition of thymocytes to cultured fetal liver cells. By contrast with the foregoing, NZB spleen cells maintained in vitro for 5 days, with or without LPS, made as much IgM during a 4-hr pulse with 3H-leucine as identically treated SWR cells. Con A-induced suppressor cells from NZB spleen cells inhibited LPS-stimulated IgM synthesis normally. There appear to be two populations of B cells in NZB mice: abnormal, spontaneously activated hypersecretors of IgM and cells that respond normally to LPS. The former population seems to arise early in life under the influence of two or three different genetic systems and functions independently of T cells.
Two Spontaneous BALB/c Lymphomas Synthesize IgM: Monomers and Half Molecules Are Isolated and Characterized Whereas Another Molecule Resembles IgD
Authors: McKeever Paul E. , Jin Kim K. , Nero Gregory B. , Laskov Reuven , Merwin Ruth M. , Logan Weltha J. , Asofsky Richard
Published: December 2022

Spontaneous lymphomas of BALB/c mice, both in vivo tumors and cell lines established in long term tissue cultures, were investigated for their ability to synthesize IgM by using radiolabeled amino acid precursors. Immunoglobulins manufactured by lymphomas K46 and L10A had the m.w. of monomeric IgM and IgM half molecule. Both of these molecules could be immunoprecipitated with class-specific anti-IgM but not anti-IgA or anti-IgG. When precipitated with polyvalent anti-Ig L10A synthesized monomeric immunoglobulins that migrated as two peaks in contrast to their single counterpart precipitated with anti-IgM. The second peak migrated in the region expected for IgD. Monomer and half molecules were composed of similar ratios of µ-chains to light chains linked by disulfide bonds. The µ2L2 monomer of these B cell lines migrated slightly slower in SDS PAGE than a µ2L2 secreted by a myeloma. Thus, these lymphomas synthesize immunoglobulins with the chemical and antigenic characteristics typical of monomeric membrane-attached IgM and IgM half molecules, plus a molecule resembling IgD on L10A only. Lymphoma assembly of monomeric IgM may follow the same initial biosynthetic sequence as myeloma assembly.
The Induction of TNP-Altered, Self-Reactive Human Cytotoxic T Cells by Soluble Factors: The Role of Ia Antigens
Authors: Friedman Steven M. , Kuhns Janet , Irigoyen Oscar , Chess Leonard
Published: December 2022

This report demonstrates that human peripheral blood T lymphocytes, triggered by allogeneic cells or soluble antigens, elaborate helper factor(s) that promotes the in vitro differentiation of TNP altered-self reactive human CTL. Helper factor(s) alone is not sufficient for the generation of these killer cells, but requires the presence of TNP-derivatized autologous stimulators during sensitization. Additional experiments were performed with antisera to a human Ia-like antigen, p23, 30. These studies indicate that human Ia-like antigens play an important role in both the induction of helper factor(s) and in the functional activity of preformed helper factor(s) molecules.
Genetic Study of Mouse Sensitivity to MHV3 Infection: Influence of the H-2 Complex
Authors: Lévy-Leblond E. , Oth D. , Dupuy J. M.
Published: December 2022

Genetic study of acute and chronic mouse hepatitis virus type 3 disease was carried out in segregating generations of a cross involving a susceptible (C57BL/6) and a resistant (A/J) mouse strain. The data obtained indicate that one or two recessive genes may be involved in resistance of acute and chronic diseases but suggest that the genes involved in both diseases are different. In this cross, no correlation was observed between H-2 and acute or chronic disease.

In mice of congenic lines, however, A/Sn (H-2a), A.SW (H-2a), A.BY (H-2b), and A.CA (H-2f), it appeared that the presence of the H-2f allele conferred to heterozygote as well as to homozygote animals the capacity to resist the development of chronic disease. It seems, therefore, that MHV3 sensitivity in mice is under the influence of at least two major genes: one for the acute disease and the other, H-2 linked, for the chronic disease.
Synergy between T Cell-Replacing Factor and Bacterial Lipopolysaccharide (LPS) in the Primary Antibody Response in Vitro: A Model for Lipopolysaccharide Adjuvant Action
Authors: Jacobs Diane M.
Published: December 2022

Unfractionated spleen cells, B cells from normal mice, and nu/nu spleen cells respond to the addition of bacterial lipopolysaccharide (LPS) and T cell-replacing factor (TRF) by production of plaque-forming cells (PFC) in excess of the number expected from the addition of LPS and TRF separately. This synergistic activity is dependent on the presence of the antigen, SRBC. Supernatants of both allogeneic spleen cell mixtures and spleen cells cultured with Con A are effective and synergize best at concentrations suboptimal for their ability to act as TRF alone. Culture supernatants of unstimulated normal or fractionated cell populations are ineffective. Synergy is not dependent on the presence of macrophages in the cultures. Purified LPS free from active contaminants, as well as commercially available LPS, show synergy with TRF. Synergy was seen when TRF was added at initiation of culture or 24 hr later. It is suggested that synergy is the equivalent of LPS adjuvant activity, that the role of T cells in LPS adjuvanticity is that of a conventional cooperating cell, and that LPS acts as an adjuvant by inducing B cells to become more sensitive to T cell helper factors.
Lipid-Modified Antigens
Authors: Janeway C. A. , Horowitz M. , Dailey M. O. , Hunter R. A. , Wigzell H.
Published: December 2022

Dodecanoic acid side chains were coupled to the antigen 2,4-dinitrophenyl-bovine serum albumin (DNP-BSA) to make DNP-BSA-lipid. T cells from guinea pigs primed with DNP-BSA-lipid proliferate in vitro when stimulated by most, but not all, DNP-carrier conjugates. Furthermore, DNP-BSA-lipid is a potent stimulator of T cells from DNP-mycobacteria primed animals. This results from its ability to stimulate virtually all DNP-reactive T cells present in such animals, whereas unmodified DNP-BSA stimulates only DNP-BSA reactive clones. When DNP-BSA-lipid was presented to T cells by first absorbing it to peritoneal exudate cells (PEC), it was found that the PEC and the immune T cells had to share I region genes for stimulation to occur. DNP groups can be shown to be present on the surfaces of DNP-BSA-lipid pulsed PEC, as determined by binding of radioactive anti-DNP antibody. Unlabeled anti-DNP will inhibit the binding of labeled anti-DNP to the pulsed PEC, but the presence of the anti-DNP in culture will not prevent stimulation of DNP-reactive T cells by the same pulsed PEC. Taken together, these data suggest that lipid modification of DNP-BSA leads to an increase in its affinity for cell surfaces. Furthermore, they may indicate that this antigen can mimic other DNP-protein antigens by means of a nonspecific association with I region gene products on the surfaces of antigen-presenting cells.
A Cell Population in nu/nu Spleen Can Prevent Generation of Cytotoxic Lymphocytes by Normal Spleen Cells against Self Antigens of the nu/nu Spleen
Authors: Miller Richard G. , Derry Helen
Published: December 2022

Spleen cells from athymic nu/nu mice contain two kinds of physically separable active cells that can have very different effects on the generation of CL (cytotoxic lymphocytes) by normal LN cells in an in vitro response against allogeneic stimulator cells. They can provide an accessory cell required for the activation of CLP (cytotoxic lymphocyte precursor cells) which need not be H-2 identical to the CLP and will function normally even when H-2 identical to the stimulator cells. They can also provide a suppressor cell that prevents the activation of CLP that can recognize the H-2 of the nu/nu mouse. Thus, with A, B, and C to represent three H-2 different mouse strains, a culture containing CLP from strain A and nu/nu spleen cells from strain B or strain (A × B)F1 will produce CL against strain C or (A × C)F1 stimulator cells but not against strain B or strain (A × B)F1 stimulator cells unless the suppressor cell is first removed. It is proposed that the in vivo role of the suppressor cell in a normal mouse is to prevent the activation of CLP reactive against self.
In Vitro Analysis of Allogeneic Lymphocyte Interaction
Authors: Delovitch Terry L. , Sohn Uik
Published: December 2022

Allogeneic effect factors (AEF) were produced across an I-J subregion incompatibility. The helper activity of these AEFs is H-2 restricted since they help B cells only of the stimulator haplotype and of other haplotypes that carry the same I-J subregion gene(s) as the stimulator haplotype. Immunoadsorption studies demonstrate that they consist of I-J determinants derived initially from the GVHR host and MLR stimulator cells and not the GVHR donor and MLR responder cells used to generate AEF. It is postulated that the genetic restriction of AEF helper activity is mediated in part by the ability of the GVHR activated donor T cells to acquire, in vivo, recipient T cell and/or macrophage derived I-J determinants. Cellular adsorption studies indicate that AEF helper activity may be adsorbed by B cells, but neither T cells nor macrophages, of the stimulator haplotype. The results suggest that an I-J-positive AEF interacts with an I-J subregion controlled complementary recognition structure on a target B cell and, after antigenic stimulation, activates that B cell to IgG antibody synthesis.
Hybrid Resistance to BALB/c Plasmacytomas
Authors: Clare Walker Mary , Phillips-Quagliata Julia M.
Published: December 2022

Genetic control of hybrid resistance to the BALB/c plasmacytoma MPC-11 was investigated. The results indicate that a single dominant autosomal gene or gene complex, which segregates independently of H-2 and the coat color c and b-loci, controls resistance to this tumor. This gene has the same strain distribution pattern in the CXB Bailey recombinant inbred strains as three unlinked genes, H-2, Ly-4, and Ea-4. It is possible, therefore, that it could be linked to either of the latter two loci. Strains that carry a positive allele for resistance are C57BL/10 and all of its congenic resistant partners tested, C57BL/6, C57L, C57BL/Ks, AKR, and DBA/1. BALB/c and its congenic resistant partners are presumed to carry a negative allele of the gene for resistance to MPC-11. Strains such as SJL, DBA/2, and A and its congenic resistant partners, which form susceptible hybrids with BALB/c, could carry either the negative allele of the gene for resistance, like BALB/c, or could carry both a positive allele of the gene and some other gene conferring susceptibility on the hybrids. Heterozygosity within the H-2 complex increases resistance only in the presence of this non-H-2 linked gene for resistance, and the effect maps to the left of the H-2D region.
Effects of Arachidonic Acid and Other Unsaturated Fatty Acids on Mitogenesis in Human Lymphocytes
Authors: Kelly James P. , Parker Charles W.
Published: December 2022

The effect of fatty acids and other lipids on mitogenic responses in cultured human peripheral blood lymphocytes was studied. Several-fold enhancement of tritiated thymidine incorporation was observed at 0.1 to 5.0 µg/ml concentrations of arachidonic acid. Other unsaturated fatty acids produced less marked changes. Increased responsiveness was demonstrable in a variety of media including RPMI 1640 supplemented with 10% fetal calf serum. Changes were also observed in uridine incorporation, total cell number, and blast transformation, indicating that the effect was not on thymidine transport or pool size per se. Arachidonic acid failed to affect PHA binding, indicating that the lectin-cell interaction was not being altered. Higher concentrations of fatty acids were inhibitory.
In Vitro Studies on the Biosynthesis of Rabbit C3: Evidence for the Synthesis of a Nascent Single Chain Precursor C3
Authors: Patel Feisal , Minta Joe O.
Published: December 2022

Biosynthesis of C3 in rabbits was studied by examining the incorporation of 14C or 3H-labeled amino acids into C3 immunoprecipitable materials by liver organ cultures and by the in vitro cell-free translation of isolated liver poly A-mRNA in the rabbit reticulocyte system. On SDS polyacrylamide gel electrophoresis, 14C-C3 immunoprecipitated from the extracellular culture medium migrated with an apparent m.w. of 200,000 daltons. In the presence of 2-mercaptoethanol it was found to dissociate into two polypeptide chains, the α chain, m.w. 130,000 and the β chain, m.w. 84,000. On the other hand, 14C-C3 immunoprecipitated from the postmitochondrial supernatant of the homogenized organ explants (intracellular C3) migrated as a single protein on SDS gels even in the presence of reducing agents.

Poly A-mRNA isolated from liver by phenol/chloroform extraction and column chromatography on oligo-(dT) cellulose was shown to direct the incorporation of 3H-leucine into TCA-precipitable protein in a concentration and time-dependent manner. Under optimal conditions the addition of exogenous mRNA resulted in a 5- to 6-fold increase in protein synthesis. The immunoprecipitated 3H-C3 migrated as a single peak of m.w. 200,000 under both reducing and nonreducing conditions. These results, taken in conjunction with the results obtained from the liver organ culture experiments, would suggest that C3 is initially synthesized as a single chain precursor protein and later modified to give the two-chain form found in serum.
Anti-DNA Activity of IgG F(ab′)2 from Normal Human Serum
Authors: Rubin Robert L. , Carr Ronald I.
Published: December 2022

The components of normal human serum (NHS) which bound DNA in a standard assay for anti-DNA antibody were characterized. IgG was the major detectable protein isolated from NHS by affinity chromatography on DNA-cellulose. A second adsorption of the whole serum IgG with DNA-cellulose did not remove additional γ-globulin indicating that only a very small fraction of the IgG was capable of binding DNA. This binding activity was largely restricted to denatured DNA. IgG (Fab′)2 bound DNA as well as the intact molecules demonstrating the antibody-like nature of the IgG binding. These results suggest that IgG antibody to denatured DNA is a normal component of human serum.
Circulating and Tissue-Bound Immune Complex Formation in Murine Malaria
Authors: June Carl H. , Contreras Carmen E. , Perrin Luc H. , Lambert Paul H. , Miescher Peter A.
Published: December 2022

Immune complex formation during Plasmodium berghei infection of OF1 mice was investigated. Circulating immune complexes (CIC) were detected by the Clq-binding assay and the conglutinin-binding solid-phase assay in lethal or drug-limited infections. CIC appeared on day 9 of infection, peaked on day 11, and disappeared only after complete cure of the infection. Analysis of the immune complexes detected by the Clq-binding assay revealed the following characteristics: sedimentation coefficients of 13S to 21S, resistance to DNAse, and selective removal by filtration through protein A bound to Sepharose. Glomerular deposits of IgM preceded the appearance of CIC, whereas deposits of IgG and C3 were concomitant with the appearance of CIC. Tissue-bound immunoglobulins were also found in the choroid plexus. The appearance of anti-malarial antibodies and malarial antigens in the serum was closely associated with a depression of C3 levels and the presence of CIC. Drug treatment was followed by normalization of C3 levels, and clearance of both CIC and malarial antigens.