bluuebunny/crossref_metadata_2025 huggingface dataset:
Specific Susceptibility of Sensitized (Memory) B Cells to Suppression and Antigenic Alteration by Murine Leukemia Virus
Authors: Cerny Jan , Waner Elizabeth B.
Published: December 2022

Mice infected with murine oncogenic RNA viruses, either Friend virus (FV) or Moloney leukemia virus (MuLV-M) reacted well to a primary immunization with antigen(s) of Vibrio cholerae; both the kinetics and magnitude of the direct (IgM) primary vibriolytic antibody plaque forming cell (PFC) response in the spleen was the same as in non-infected control mice. In contrast, the anamnestic PFC (IgM) response of mice preimmunized with cholera and later (2 to 9 months following priming) infected with either FV or MuLV-M, before the secondary challenge, was inhibited 90% or more when compared to the anamnestic response of controls. Hemolytic PFC response to a single (primary) immunization with sheep red blood cells (sRBC, an antigen to which laboratory mice have a natural immune background) was suppressed by infection with both viruses in a manner similar to the suppression of the secondary vibriolytic response.

The suppressive effect of FV, and the lack of it, on these different immune responses was correlated with the appearance of FV-induced cell membrane antigen, FVMA, on respective individual PFC from the spleen. FVMA was detected by inhibition of PFC in the presence of specific antiserum, anti-FVMA, and complement (C).

In particular, about 80% of the FV-infected mice with an anamnestic response to a secondary challenge with cholera antigen had a significant decline in the proportion of PFC (33% or more) following incubation with anti-FVMA plus C. A similar degree of inhibition was found with hemolytic (anti-sRBC) PFC from FV-infected mice. In contrast, primary vibriolytic PFC from FV-infected mice were not inhibited by anti-FVMA, i.e., they did not carry the virus-induced antigen.

These results indicate absolute correlation between viral suppression of a given clone of immunocompetent cells, and antigenic alteration of individual antibody-forming cells due to the virus infection. Furthermore, the absence of both functional inhibition and virus-induced antigen(s) in the primary immunocompetent cells and the presence of both these virus-related functions in the memory cells (either experimentally sensitized cholera memory cells or naturally sensitized sRBC memory cells) strongly suggested that the latter are selectively susceptible to oncornavirus.
Affinity Chromatography of Human Leukocyte and Diploid Cell Interferons on Sepharose-Bound Antibodies
Authors: Berg Kurt , Ogburn Clifton A. , Paucker Kurt , Mogensen K. Erik , Cantell Kari
Published: December 2022

Interferons produced in human peripheral leukocytes (LE) and foreskin fibroblast (FS-4) cells were subjected to affinity chromatography on Sepharose-bound globulins from rabbits immunized with these interferons. Anti-LE interferon sera neutralized both interferons, but titers against FS-4 interferon were consistently lower than those against LE interferon. Anti-FS-4 interferon sera neutralized only FS-4 but not LE interferon. Accordingly, affinity columns constructed with anti-FS-4 globulin excluded LE but not FS-4 interferon, whereas those prepared with anti-LE interferon globulin bound and eluted both LE and FS-4 interferons. Purification of native interferons of both types on anti-LE interferon-Sepharose ranged from 680- to 3,600-fold and recoveries from 72 to 126%. Specific activities of eluate pools varied from 4 to 30 × 106 reference (B, 69/19) units per milligram protein.
Cleavage of the Third Complement Component (C3) and Generation of the Spasmogenic Peptide, C3a, in Human Serum Via the Properdin Pathway: Demonstration of Inhibitory As Well As Enhancing Effects of Epsilon-Amino-Caproic Acid
Authors: Vogt Walther , Schmidt Gisa , Lynen Rainer , Dieminger Lothar
Published: December 2022

Human and guinea pig serum loses hemolytic activity of the third complement component (C3) during incubation at 37°C. The loss is due to specific C3 cleavage involving the properdin system. This is concluded from the finding that C3 inactivation is prevented by EDTA, by elimination of properdin factor B, and unimpaired in C4 deficient guinea pig serum.

In the presence of 1 M ε-amino-caproic acid (EACA) spontaneous C3 cleavage is considerably enhanced and accompanied by the appearance of biologically active C3a. Lower concentrations of EACA inhibit rather than enhance C3 cleavage in serum. The inhibitory effect of EACA is due to interference with the interaction of the properdin factors and their action on C3 as demonstrated in various systems: the assembly of an active C3-cleaving complex on zymosan, its regeneration after decay by factor D and factor B (GBG), cleavage of GBG by C3b and factor D in systems of purified components, and cleavage of C3 by preformed properdin complexes. Reactions involving the cobra venom factor were likewise depressed. In all these systems EACA was inhibitory even at 1 M concentration. No single step in the development or action of an active properdin system was found to be enhanced by 1 M EACA. The enhancing effect of high concentrations of EACA on C3 cleavage in serum may be explained by its observed inhibition of C3b inactivator (C3bINA). This factor controls the properdin system by destroying C3b. In serum the inhibitory effect of 1 M EACA on C3bINA appears to allow escape of the properdin system from its control and thus to increase its net activity toward C3 despite inhibition of the enzymic reactions proper. At lower concentrations the effect of EACA on C3bINA is apparently less significant; therefore, at low concentrations of EACA, its inhibitory effects on C3 cleavage by the properdin system in serum predominate.
In Vitro Response of Mouse Spleen Cells to the Solid Phase Immunogen DNP-O-Bio-Gel
Authors: Trump Gary N.
Published: December 2022

The in vitro immunogenicity of the solid-phase hapten, dinitrophenyl-ornithine-Bio-Gel (DNP-O-Bio-Gel), was investigated in cultures of mouse spleen cells. Appropriate combinations of cells and immobilized hapten were determined. Large numbers of direct anti-hapten plaque-forming cells (PFC) were generated when 1 × 107 C57BL/6 or C57BL/10 spleen cells were cultured with 4 × 103 DNP-O-Bio-Gel beads. Specificity studies of the responses of cultured spleen cells to DNP-O-Bio-Gel yielded the following results: soluble DNP-ornithine or DNP-bovine γ-globulin inhibited the induction of anti-hapten PFC by DNP-O-Bio-Gel; neither dinitrophenyl-Bio-Gel (DNP-Bio-Gel) nor ornithine-Bio-Gel (O-Bio-Gel) induced anti-hapten responsiveness; furthermore, neither DNP-Bio-Gel nor O-Bio-Gel inhibited the induction of PFC by DNP-O-Bio-Gel. It was concluded, from the results of these specificity experiments, that a spacer, ornithine, is required for immunogenicity of immobilized DNP; and that the Bio-Gel bead, itself, acts solely as a physical carrier for the hapten.
The Interferon Refractory State
Authors: Borden Ernest C. , Prochownik Edward V. , Carter William A.
Published: December 2022

When L929 cells were treated with samples collected from cells or mice producing interferon, almost complete refractoriness to subsequent interferon induction was observed. The relationship of this refractoriness-inducing product (RIP)5 to the antiviral activity of the samples was determined. Using interferon in the pretreatment sample as a measure of RIP concentration, a semilog plot of the pretreatment interferon titer and interferon subsequently produced, resulted in an approximately linear relationship between 10 and 100 units of interferon in the pretreatment sample. This relationship was largely independent of the multiplicity of Newcastle disease virus (NDV) used to produce interferon in the treated monolayers.

A 2- to 6-fold temporal differentiation in the refractoriness-inducing activity and the antiviral activities was obtained both in vitro and in vivo. Supernatants collected 48 hr after NDV infection of L929 cells had 2- to 4-fold more refractoriness-inducing activity than those collected earlier. In serum of ICR mice given poly rIn:rCn-poly-d-lysine, a quantitatively similar temporal increase in refractoriness-inducing activity was observed. Treatment of L929 cells with cycloheximide resulted in a 4-fold increase in interferon production. At equivalent interferon concentrations, supernatants from the cycloheximide-treated monolayers inhibited interferon production 4-fold less than did comparable supernatants from non-cycloheximide-treated monolayers. Soluble cell extracts inhibited interferon production to a greater degree than did extracellular preparations. The protein nature of the refractoriness-inducing activity was suggested by its trypsin sensitivity. These findings suggest that the refractoriness-inducing and antiviral activities of interferon preparations result from different proteins.
Thyroglobulin-Binding Lymphoid Cells in Obese Strain (OS) Chickens
Authors: Richter E. , Wick G.
Published: December 2022

The binding of chicken thyroglobulin-coated chicken red blood cells by splenic, thymic, and bursal lymphoid cells was analysed in Obese strain (OS) chickens with spontaneous autoimmune thyroiditis and normal white Leghorn controls aged 1 week to 2.5 yr. Chicken erythrocytes coated with pneumococcal polysaccharide SIII served as controls. The specificity of thyroglobulin-rosette-forming cells was verified by inhibition experiments. OS chickens showed significantly higher counts of thyroglobulin-rosette-forming cells in the spleen and thymus as compared to normal controls while no such difference was found for SIII-rosette-forming cells. The values obtained with bursal lymphoid cells were in the same range in both OS and normal white Leghorn birds. The appearance of thyroglobulin-rosette-forming cells and their peak values clearly preceded the maximum frequency and severity of thyroditis and the peak of the thyroglobulin antibody curve. It is concluded that the presence of thyroglobulin rosette-forming cells is a prerequisite for the future development of spontaneous autoimmune thyroiditis in the OS. Inhibition studies with specific turkey anti-chicken bursa and thymus cell sera revealed the B cell nature of active thyroglobulin-rosette-forming cells and suggested that passive rosette-forming cells were of T cell origin.
Rat Cell Surface Antigens
Authors: Callahan Gerald N. , Dewitt Charles W.
Published: December 2022

A cell surface antigen was isolated from soluble extracts of rat lymphoid cells. Molecular weight of the purified antigen was estimated by SDS gel electrophoresis to be 10 to 20,000 daltons. Physical as well as immunologic properties of this antigen do not resemble those of any known antigenic product of the major histocompatibility locus of the rat. It is therefore proposed that it is a rat minor histocompatibility antigen.
The Suppressive Effect of Immunization on the Proliferative Responses of Rat T Cells in Vitro
Authors: Bash Jerry A. , Waksman Byron H.
Published: December 2022

DNA synthesis (3H-thymidine incorporation) of DA rat spleen and lymph node cells cultured with PHA or (DA × Lewis)F1 lymph node cells was profoundly inhibited by pretreatment with large i.v. doses of antigen (τ;100 mg OA or τ; 1010 SRBC) 24 hr before culture. This effect, seen as early as 3 to 6 hr after antigen administration, had disappeared within 7 days. Similar depressed spleen cell responses were seen 9 days after footpad injection of 200 µg OA in complete adjuvant. Lymph node responses showed a similar inhibiton only in the presence of specific antigen in culture. In all experiments in which antigen administration resulted in decreased responses, removal of glass wool adherent cells restored responsiveness to normal. The failure of purified macrophages to reestablish inhibition favored the interpretation that an adherent suppressor cell distinct from the macrophage is activated by antigen in vivo to regulate T cell responses in vitro.
The Modulation of Spontaneous and Anti-Ig-Stimulated Motility of Lymphocytes by Cyclic Nucleotides and Adrenergic and Cholinergic Agents
Authors: Schreiner George F. , Unanue Emil R.
Published: December 2022

Translational movement in B lymphocytes was stimulated by anti-Ig antibody. Drugs that are presumed to elevate cyclic AMP stopped this stimulated motility. Such was the case with dibutyryl cAMP and theophylline, cholera enterotoxin, and isoproterenol, a β adrenergic agonist. Conversely, in the absence of anti-immunoglobulin antibody, cyclic GMP and the cholinergic drugs acetylcholine and carbamylcholine increased spontaneous motility of lymphocytes, with the B class of lymphocytes demonstrating greater responsiveness. The increase in motility brought about by cholinergic drugs was totally stopped by atropine, suggesting that the B lymphocyte surface contains a cholinergic receptor.

The inhibition of anti-immunoglobulin-stimulated movement produced by cyclic AMP was not observed if the cells were first incubated with colchicine, the microtubular-disrupting drug. This suggests that the cyclic AMP-decreased motility was brought about by the increased stabilization of microtubules. Lymphocyte motility was dissociable from other early events subsequent to binding of anti-immunoglobulin antibodies: patching, capping, and endocytosis of complexes were unaffected by cyclic AMP, cyclic GMP, or drugs of the adrenergic or cholinergic systems.
Identification of a Colon-Specific Antigen (CSA) in Normal and Neoplastic Tissues
Authors: Goldenberg David M. , Pegram Charles A. , Vazquez Jacinto J.
Published: December 2022

An antigen has been isolated from a human signet-ring cell carcinoma serially growing in hamsters, GW-39, by saline, PCA, or phenol extraction, and has been found immunologically identical to a similarly extracted substance in normal human or hamster colon. No other hamster or human tissues or cells were found to contain this antigen, for which reason we have termed it colonspecific antigen, or CSA. CSA has been found to be distinct from the major blood group-specific antigens and from other colon tumor-associated antigens, such as CEA, CCA-II, and CCA-III. It thus seems that a colon organ-specific antigen can be synthesized by this particular human tumor system.

Hamsters immunized with CSA could reject cheek pouch grafts of GW-39 tumors, and tumor rejection by these animals correlated with their anti-CSA antibody titers. Preliminary characterization of CSA suggested that it is a glycoprotein on the cell surface having a molecular size of 30,000 to 50,000 daltons. It is proposed that CSA may play a role in the diagnosis of mucin-producing adenocarcinoma of the colon and in ulcerative colitis.
Specific Stimulatory and Cytotoxic Effects of Lymphocytes Sensitized in Vitro to Either Alloantigens or Tumor Antigens
Authors: Kall Mary Ann , Hellström Ingegerd
Published: December 2022

Lymph node cells (LNC) or spleen cells from normal BALB/c mice were sensitized in vitro by co-cultivation with mitomycin C-treated allogeneic sarcoma cells, and their reactivity measured with a microcytotoxicity assay. Following 5 days of sensitization, a specific cytotoxic effect against tumor cells carrying the sensitizing alloantigens was observed; this is in agreement with previously published findings. Lymphoid cells tested after 3 days of sensitization, however, specifically increased the number of target cells remaining in the Microtest plates at the conclusion of the microcytotoxicity assays.

LNC from normal BALB/c mice were also sensitized in vitro against tumor specific antigens using four different methylcholanthrene-induced sarcomas of BALB/c origin. After 6 days of sensitization the LNC were specifically cytotoxic for the immunizing tumor cell line. After 3 days of sensitization, however, the LNC gave a specific increase of the number of remaining target tumor cells.
Studies on the Synthetic Capacity and Antigenic Expression of Glutaraldehyde-Fixed Target Cells
Authors: Bubbers J. Eric , Henney Christopher S.
Published: December 2022

Mastocytoma cells (P815 of the DBA/2 strain) treated with increasing concentrations of glutaraldehyde were concurrently evaluated for their ability to incorporate exogeneous uridine, thymidine, and amino acids. The antigenic expression and membrane integrity of these treated cells were assayed by measuring their susceptibility to lysis by antibody and complement and by T-effector cells.

The concentrations of glutarladehyde required to affect target cell antigen display were greater than those required to inhibit totally the cell's protein and nucleic acid synthetic processes. Thus, P815 cells treated with 0.15% glutaraldehyde for 10 sec were unable to incorporate either amino acids or nucleotides into macromolecules, but were readily lysed by effector T cell populations, and by alloantibody in the presence of complement. Target cells treated with glutaraldehyde concentrations in excess of 0.25% for 10 sec were resistant to both forms of immune lysis.

In keeping with these observations, monolayers of P815 cells treated for 10 sec with 0.15% glutaraldehyde, were capable of specifically depleting T-effector cells from a cytolytically-active spleen cell population. After treatment with higher concentrations of glutaraldehyde (0.3%), however, the monolayers lost their capacity to adsorb effector cells.

Although P815 cells treated with glutaraldehyde continued to exhibit H-2d alloantigen, neither these cells nor glutaraldehyde-treated DBA/2 spleen cells induced significant blastogenesis or stimulated the production of cytolytically active effector cells in mixed leukocyte cultures.
Detection of Cell-Bound Immunoglobulins by a Radioisotopic Micro-Mixed Hemadsorption Reaction with Technetium-99m-Labeled Erythrocytes
Authors: Wood Gary W. , Barth Rolf F.
Published: December 2022

The mixed hemadsorption (MHA) reaction detects antibodies reactive with cell surface antigens by means of antiglobulin-coated indicator erythrocytes. We have developed a radioisotopic modification which employs sheep erythrocytes (SRBC) that have been prelabeled with technetium-99m (99mTc), a high specific activity metastable γ-emitter of short half life. The 99mTc MHA reaction was performed on human and murine cells cultured in Microtest II plates with six replicate wells per serum dilution. Antibody activity in species-specific xenoantisera and mono- and polyspecific alloantisera was detected in high titer. The sensitivity of 99mTc micro-mixed hemadsorption was 2 times that of the visual assessment of mixed hemadsorption, 100 to 200 times that of the 125I-mixed antiglobulin reaction and 500 to 1000 times more sensitive than indirect immunofluorescence. The assay system was applied successfully to confirm the species of origin of a panel of previously karyotyped human and mouse cell lines. Our results indicate that the 99mTc micro-mixed hemadsorption method is a rapid, sensitive, quantitative test for the detection of cell surface antigens and membrane reactive antibodies.
Receptor Sites for Antigen-Antibody Complexes on Cells Derived from Solid Tumors: Detection by Means of Antibody Sensitized Sheep Erythrocytes Labeled with Technetium-99m
Authors: Wood Gary W. , Gillespie G. Yancey , Barth Rolf F.
Published: December 2022

Surface receptor sites for the Fc portion of antigen-antibody complexes were demonstrated on cells derived from three methylcholanthrene-induced fibrosarcomas, one of strain C3H and two of strain BALB/c origin, two spontaneously occurring malignant melanomas (B16 in strain C57BL/6 and Harding-Passey in strain BALB/c mice), a Moloney sarcoma virus-induced tumor of strain BALB/c origin and the Walker 256 carcinosarcoma of Holtzman rats. Primary cell cultures derived from these tumors adsorbed technetium-99m labeled, antibody-sensitized sheep erythrocytes (99mTc EA) as determined either by visual scoring of adherence or radioisotopic quantitation. Depending upon the tumor tested, from 20% to greater than 95% of the target cells absorbed 99mTc EA. All cells lost their reactivity after 1 or 2 passages in vitro, but this was regained after a single passage in vivo. Indicator erythrocytes coated with F(ab′)2 fragments of the sensitizing sheep erythrocytes (SRBC) antiserum did not adhere thereby demonstrating that the hemadsorption required an intact Fc portion of the antibody molecule. Adherence of 99mTc EA was blocked by soluble immune complexes prepared with ovalbumin and rabbit antibody directed against it and Escherichia coli 055:B5 lipopolysaccharide and mouse antibody directed against it. Normal rabbit or mouse serum, immune serum, or antigen alone did not block adherence of 99mTc EA thereby demonstrating that the receptors had greater affinity for immune complexes than for either antigen or antibody alone. The existence of membrane receptors on tumorderived cells which react with the Fc portion of antigen-antibody complexes may provide an explanation for the mechanism by which immune complexes are capable of blocking cell-mediated tumor cell destruction irrespective of whether the receptors are on the tumor cells themselves or on admixed lymphocytes and macrophages.
Immune Response to Liposomal Model Membranes: Restricted IgM and IgG Anti-Dinitrophenyl Antibodies Produced in Guinea Pigs
Authors: Uemura Kei-Ichi , Claflin J. Latham , Davie Joseph M. , Kinsky Stephen C.
Published: December 2022

Guinea pigs were immunized with liposomal model membranes actively sensitized with 2,4-dinitrophenyl-aminocaproylphosphatidyl-ethanolamine. The immune response was characterized by the formation of both IgM and IgG anti-DNP plaque-forming cells (PFC) which paralleled the appearance of anti-DNP antibodies in the serum. Plaque inhibition by DNP-lysine indicated that the PFC produced after liposomal immunization fell in a much narrower range of avidity groups than the PFC obtained after immunization with DNP-albumin and, in this regard, resembled MOPC 315. The restricted nature of the serum IgG anti-DNP antibodies was confirmed by isoelectric focusing and revealed the expression of a limited number of clones in all cases.
Impaired Macrophage Function in Reiend Virus Leukemia: Restoration by Statolon
Authors: Levy Michael H. , Wheelock E. Frederick
Published: December 2022

Phagocytic and migratory functions of peritoneal macrophages from Friend virus (FV) leukemic mice are significantly depressed as compared with normal controls. Leukemic macrophages exposed in vivo and in vitro to statolon, an extract of the mold Penicillium stoloniferum, shown previously to suppress FV erythroleukemia, regain normal function and release reduced amounts of FV. Statolon's in vivo restoration of leukemic macrophage function is paralleled by restoration of humoral immune competence. Statolon induces interferon in vitro but its effects on leukemic macrophages are probably direct, since restoration of macrophage function occurs at dosage levels far below those that induce interferon. These studies suggest that macrophages play an integral role in both the pathogenesis and the statolon-induced suppression of FV disease.
Partial Characterization of Leukocyte Inhibitory Factor by Concanavalin A-Stimulated Human Lymphocytes (LIFCon A)
Authors: Rocklin Ross E.
Published: December 2022

Leukocyte inhibitory factor (LIF) obtained from normal human lymphocytes stimulated by Concanavalin A was characterized by Sephadex gel filtration, disc electrophoresis, isopycnic centrifugation, enzymatic treatment and by immunoabsorbent studies. LIF activity was found after Sephadex G-100 gel filtration in a fraction containing molecules having the size of albumin (68,000 daltons). This is the same region where LIF activity derived from antigen stimulated lymphocytes has previously been found to elute. On disc gel electrophoresis at pH 9.1, peak LIF activity was eluted from the acrylamide gel fraction containing molecules that migrate with albumin. In isopycnic centrifugation studies, human LIF was shown to have a buoyant density similar to that of pure protein. Human LIF was inactivated by treatment with chymotrypsin but not by neuraminidase treatment. LIF activity could not be removed by incubating the material on columns containing Sepharose 6b coupled with anti-human serum albumin or anti-Fab. The characteristics of human LIF and migration inhibitory factor were compared.
IgG on Lymphocyte Surfaces; Technical Problems and the Significance of a Third Cell Population
Authors: Winchester R. J. , Fu S. M. , Hoffman T. , Kunkel H. G.
Published: December 2022

Direct immunofluorescence performed with the F(ab′)2 fragment of rabbit antibodies to IgG revealed that membrane bound IgG was only rarely found on the surface of small peripheral blood lymphocytes (PBL). In contrast whole antibodies to IgG used in fluorescence gave much higher levels of cells with IgG surface staining. This staining resulted from secondary IgG binding, in part due to the uptake of newly formed immune complexes. IgM-and IgD-bearing cells were brightly stained in relatively similar percentages by both the whole and F(ab′)2 forms of the class-specific antibodies; they constitute the principal membrane Ig of PBL.

Evidence was obtained indicating that a special population of cells with Fc receptors but lacking membrane Ig was primarily involved in the high IgG binding. This population also formed sheep erythrocyte rosettes when optimal conditions were utilized.
Antibody-Mediated Suppression of Grafted Lymphoma
Authors: Shin Hyun S. , Hayden Michael , Langley Stephen , Kaliss Nathan , Smith Mary Ruth
Published: December 2022

Syngeneic or allogeneic mice pretreated with sublethal whole-body irradiation were rendered incapable of suppressing the growth of grafted tumor cells sensitized with alloantibody. The growth of sensitized tumor cells was suppressed when they were mixed with donor effector cells from mice syngeneic or allogeneic to the recipients and then were inoculated in irradiated recipients. Three donor-host combinations were used to study the suppression of the murine lymphoma 6C3HED indigenous to C3H mice. These were C3H donor cells in C3H recipients, C57BL/6 donor cells in C3H recipients, or C57BL/6 donor cells in C57BL/6 recipients. In all three combinations, macrophages obtained from an inflammatory exudate, exudate lymphocytes not bearing θ antigen, and platelets were, in descending order of effectiveness, consistently active in restoring antibody-mediated suppression of tumor growth in irradiated hosts. Prior irradiation of the transferred lymphocytes somewhat diminished their effectiveness. Freeze-thawed or heat-killed macrophages (but not freeze-thawed platelets or lymphocytes) were effective in restoration. Peripheral blood mononuclear leukocytes and splenic lymphoid cells were not active in the recipients syngeneic to the donor cells but were active in recipients allogeneic to the donor cells. Polymorphonuclear leukocytes isolated from peripheral blood or an inflammatory exudate were not active. Intact thymic function seems unimportant since antibody-mediated suppression took place as effectively in thymectomized mice as in normal controls.
Studies on the Control of Antibody Synthesis
Authors: Kim Young Tai , Greenbaum Donald , Davis Paul , Fink Steven A. , Werblin Theodore P. , Siskind Gregory W.
Published: December 2022

The effect of boosting on antibody affinity was studied in a haptenic system. Generally, boosting results in the prompt synthesis of high affinity anti-hapten antibody. However, repeated boosting frequently leads to a decrease in the amount and affinity of the serum antibody. Repeated boosting with hapten on a carrier different from that used for priming selectively stimulates synthesis of the highest affinity anti-hapten antibody and does not result in a decrease in affinity. Priming with soluble antigen without adjuvants results in the synthesis of low affinity antibody. After such priming, boosting stimulates low affinity antibody synthesis and repeated boosting leads to a moderate increase in antibody affinity.
The Allogeneic Effect: M-Locus Differences Substitute for Differences in the H-2 Major Histocompatibility Complex
Authors: Röllinghoff Martin , Wagner Hermann
Published: December 2022

The primary immune response against sheep red blood cells in T cell-deficient spleen cell cultures from nude mice was tested in the absence and presence of allogeneic spleen cells. The allogeneic spleen cells differed either in regard to the major histocompatibility complex (H-2) or only with respect to the M-locus. Surprisingly the M-locus different spleen cells were almost as efficient in enhancing the anti-sheep red blood cell response in nude cultures as were the cells differing on the complete H-2 complex. Evidence is presented that AKR anti-θ serum sensitive T cells are responsible for the M-locus-dependent effect described. This effect is shown to be mediated by a factor released from actived T cells stimulated in M-locus different mixed lymphocyte cultures. Since almost identical parameters have been observed both in the M-locus-dependent situation as in the “classical” allogeneic situation we concluded that an allogeneic effect can be induced by T cells responding to a complete set of the major histocompatibility complex (H-2) as well as to lymphocyte-activating determinants (M-locus) alone.
Transplantable Immunoglobulin-Secreting Tumors in Rats
Authors: Querinjean Pierre , Bazin Hervé , Kehoe J. Michael , Capra J. Donald
Published: December 2022

Most of the spontaneous ileocecal immunocytomas of LOU/C/Wsl strain rats secrete large amounts of monoclonal immunoglobulins. Amino acid sequence studies were undertaken on the heavy chains of 13 of these proteins and on pooled heavy chains from LOU rats to define the structural variability in the amino terminal region of the rat heavy chain. Characteristic VHIII sequences were found in both the myeloma proteins and the heavy chains from the pool. The data confirm previous conclusions concerning the rat VHIII subgroup that had been derived from studies of pooled heavy chains from Sprague-Dawley rats. In particular phylogenetically associated residues, identical to those previously determined in the Sprague-Dawley pool, were found in the LOU/C/Wsl rat myeloma proteins.

There were two unexpected findings: 1) hypervariability at position 16 of the rat VHIII myelomas as contrasted with more than 90% glycine in the pool, and 2) a striking homology of one rat myeloma heavy chain with the variable region of the MOPC 315 mouse α-chain.

The findings show that the rat myeloma system provides useful proteins for sequence analysis but indicate that some sequence pattern differences can be detected between the myeloma proteins and pooled immunoglobulins from this species.
Fractionation of Functional Lymphocytes Sensitized to Basic Encephalitogen on Derivatized Collagen and Gelatin Gels
Authors: Webb Cynthia , Teitelbaum Dvora , Rauch Helene , Maoz Arieh , Arnon Ruth , Fuchs Sara
Published: December 2022

The lymph node cells of basic encephalitogen (BE)-sensitized guinea pigs were fractionated on derivatized collagen and gelatin gels. The population of cells specifically reactive to this antigen can be isolated from derivatized gelatin gels and retain their viability and functionality as assayed in vitro. The specific binding of BE-sensitized cells to BE-derivatized gels comprised between 1 and 2% of the cells applied per plate. The ratio of sensitized cells bound to non-sensitized cells bound ranged between 4 and 6. The viability and functionality of adherent cells detached from collagen gels after enzymatic degradation were impaired. In contrast, the responses obtained with the adherent cell population released from the gelatin gels, by melting at 37°C, were equal or greater than those of the original unfractionated lymph node cell cultures. Furthermore, it was possible to obtain a nonadherent cell population which was virtually completely depleted of the capacity to respond to the sensitizing antigen.
Modulation of Cyclic AMP in Purified Rat Mast Cells
Authors: Sullivan Timothy J. , Parker Keith L. , Eisen Seth A. , Parker Charles W.
Published: December 2022

Changes in intracellular and extracellular rat mast cell adenosine 3′:5′ monophosphate (cAMP) concentrations during stimulation of histamine release by 48/80 were studied. There was a rapid and progressive fall in intracellular cAMP beginning within 10 sec after the addition of 48/80. The lowest cAMP values were obtained at 10 min, with return to control levels by 30 min. The fall in cAMP was dose-related with progressive decreases in 10-min cAMP measurements as the 48/80 concentration was increased from 0.25 to 1.00 µg/ml. There was a graded increase in histamine release over the same concentration range. Attempts to demonstrate significant amounts of cAMP in the medium during 48/80 stimulation were unsuccessful, indicating that the changes in cAMP intracellularly are not due to altered cellular permeability. There was a general correlation between the ability of pharmacologic agents to sustain high intracellular levels of cAMP in the presence of 48/80, and inhibition of histamine release. Theophylline (20 mM) which increased cAMP levels 2-3-fold prevented a detectable decrease in cAMP after 1 µg/ml 48/80 (measured at 10 min) and almost completely inhibited histamine release. Prostaglandin E1 (27 µM) also raised cAMP levels, decreased the 48/80-induced fall in cAMP (by 31%), and partially inhibited histamine release (by 42%). Epinephrine increased mast cell cAMP levels, but did not prevent the subsequent 48/80-induced decrease in cAMP and did not inhibit histamine release. Carbamylcholine (1 nM), adenine (1 µM), and diazoxide (10 µM) lowered mast cell cAMP and potentiated 48/80 induced release.

In view of previous studies from this laboratory indicating that 48/80 stimulates mast cell phosphodiesterase, it seems likely that the 48/80-induced fall in cAMP is due, at least in part, to increased cAMP destruction. Since agents which prevent the fall in cAMP inhibit histamine release, it is apparent that cAMP is an important part of the control mechanism of histamine secretion. On the other hand, it cannot be concluded that a decrease in cAMP alone is sufficient to produce a response since carbamylcholine, diazoxide, and adenine which lower cAMP do not alter histamine release unless 48/80 is also present.
The Induction of Delayed Hypersensitivity in Guinea Pigs to Poly U and Poly A:U
Authors: Casavant Conrad H. , Youmans Guy P.
Published: December 2022

Guinea pigs were sensitized to poly U and poly A:U so that subsequent stimulation of spleen cells from these immunized animals with poly U and poly A:U resulted in the production of migration inhibitory factor (MIF). MIF was also produced when spleen cells from animals immunized with poly A:U were cultured in the presence of mycobacterial RNA or whole viable H37 Ra cells. Negative dermal reactions were observed when guinea pigs immunized with poly A, poly U, or poly A:U were skin tested with these same synthetic nucleotides.
Properties of Conjugated Protein Immunogens Which Selectively Stimulate Delayed-Type Hypersensitivity
Authors: Coon John , Hunter Robert
Published: December 2022

If bovine serum albumin (BSA) is covalently conjugated with dodecanoic acid, its ability to stimulate delayed-type hypersensitivity (DTH) is enhanced whereas its ability to stimulate antibody production is suppressed. Conjugates of BSA prepared with negatively charged succinyl groups and positively charged amidine groups were unable to stimulate any detectable DTH to BSA even though their ability to bind anti-BSA antibody in a radioimmunoassay was similar to that of dodecanoyl-BSA which stimulated strong, sustained DTH. By analyzing a series of conjugates of BSA with fatty acids of varying chain length, we found that the enhancement of the ability of a preparation to stimulate DTH to BSA could be separated from the suppression of its ability to stimulate antibody formation. The ability of a preparation to stimulate DTH correlated with the covalent conjugation of a large number of hydrophobic groups whereas the suppression of the ability to stimulate antibody formation correlated with covering of antibody-combining sites as measured in a radioimmunoassay.
Preparation and Characterization of Rabbit IgG Fractions
Authors: Schlamowitz Max , Kaplan Morris , Shaw Anita R. , Tsay David D.
Published: December 2022

Rabbit IgG has been purified and further fractionated by DEAE-cellulose column chromatography to yield three fractions, Fr-I, -II and -III-1 that span a broad range in the heterogeneous molecular population of IgG. Immunodiffusion established that these fractions are free from contamination by IgA, IgM, transferrin, and hemopexin. The three fractions were assayed for neutral hexose and sialic acid; Fr-III-1 and Fr-I yielded the highest and lowest values for these components, respectively. The order of elution from DEAE-cellulose and the electrophoretic mobilities of the fractions correlate with their content of sialic acid. Digestion of Fr-I with papain is much more rapid and complete than that of Fr-III-1. Treatment with neuraminidase to remove sialic acid from each fraction does not affect the comparative susceptibility to digestion by papain. The fractions can also be distinguished by the relative amounts of H-L half molecules and H chains formed under identical conditions of reduction with cysteine and by differences in their ability to bind with IgG receptors on the fetal rabbit yolk sac membrane.

The results are discussed in the context of the structures of IgG in the heterogeneous population of molecules and the possible relation to the oligosaccharide moieties of the IgG molecules in that population.
Alkaline Phosphatase in Differentiating Guinea Pig Thymocytes: An Ultracytochemical Study
Authors: Ruuskanen Olli J. , Pelliniemi Lauri J. , Kouvalainen Kauko E.
Published: December 2022

Alkaline phosphatase (AP) was demonstrated ultracytochemically on the cell membrane of guinea pig thymocytes. AP usually covers the whole cell surface. In a small subpopulation of thymocytes the cell membrane is totally AP-negative. These cells are located by light microscopy in the thymic medulla and have been shown previously to possess the migratory properties of mature thymocytes. This AP-negative subpopulation most probably represents the T cell population in the thymic medulla. No distinct morphologic differences could be observed between these AP-positive and AP-negative thymocytes. Additionally, a small AP-negative cell population with characteristics of immature lymphoid cells was noted. These lymphoid cells, which are seen more numerously in the fetal thymus, might represent the differentiating stem cells in the guinea pig thymus.
Cell Surface Immunoglobulin. XIII. Distribution of IgM and IgD-Like Molecules on Small and Large Cells of Mouse Spleen
Authors: Goodman S. A. , Vitetta Ellen S. , Melcher Ulrich , Uhr Jonathan W.
Published: December 2022

The distribution among murine spleen cells of a newly described class of surface immunoglobulin (Ig) with properties similar to human IgD was studied. Splenocytes were separated on the basis of size and the surface Ig on large cells (sedimenting faster than 6 mm/hr in a 1 × G velocity gradient) and small cells (sedimenting between 2.5 and 3.0 mm/hr) was analyzed. Spleen cells from young animals had virtually only IgM on the large cells but had substantial amounts of IgM and the IgD-like molecule (IgD) on small cells. Spleen cells from older animals, which have larger amounts of IgD, had IgM and IgD on both cell types; however, the amount of IgD relative to IgM on the large cells was always substantially less than that on the small ones. These observations taken together with those of other investigators support the hypothesis that a large lymphocyte with surface IgM is the precursor of a small lymphocyte with both surface IgM and IgD.
Immunochemical and Chemical Studies on Streptococcal Group-Specific Carbohydrates
Authors: Coligan John E. , Schnute William C. , Kindt Thomas J.
Published: December 2022

Structural studies on the carbohydrates of Groups A, C, and A-variant (AV) streptococci have utilized periodate oxidation, permethylation analysis, and immunochemical comparison of intact and periodate-oxidized polysaccharides. The data indicate that a similar 1,2- and 1,3-linked rhamnose chain is present in both the A and AV carbohydrates. The group A carbohydrate contains in addition N-acetylglucosamine residues at nonreducing terminals, whereas the AV is a homopolymer of rhamnose. There is some evidence that Group C carbohydrate contains the same rhamnose chain, but structural comparisons to the A and AV carbohydrates are complicated by the presence of intrachain N-acetylgalactosamine residues. Periodate oxidation and permethylation analysis show that while approximately 50% of the N-acetylgalactosamine of the Group C carbohydrate occupies terminal positions, the remainder is present as 1,3-linked units. Removal of the nonreducing terminal hexosamine units from the Group A carbohydrate by periodate treatment significantly enhanced its cross-reactivity with AV antiserum, whereas no enhancement was observed after similar treatment of the Group G carbohydrate. The data indicate the presence of an α-1,3-linked N-acetylgalactosamine disaccharide at the nonreducing terminal of the Group C carbohydrate.
Control of Proliferation in Mitogen Responsive Human Peripheral Blood Lymphocytes: Restimulation of Cells Stimulated by Sodium Periodate
Authors: Norin Allen J. , Strauss Bernard S.
Published: December 2022

Human peripheral blood lymphocytes are stimulated to a greater extent by sodium periodate when cells are incubated in medium containing human serum than when incubated in medium with fetal calf serum. NaIO4 stimulation can be reversed by treatment with sodium borohydride but cells already committed to division are not affected by borohydride treatment. Maximal commitment to DNA synthesis of a NaIO4 oxidized cell suspension occurs after about 28 hr of incubation in medium. The committal time after periodate stimulation is identical to that after stimulation with concanavalin A. Cells treated with periodate and then reduced with borohydride immediately after oxidation are refractory to further periodate stimulation. Cells stimulated with periodate and then incubated for 6 hr before treatment with borohydride can be restimulated with periodate, indicating a turnover of membrane sites in the 6 hr period. Periodate-stimulated cells divide only once in response to the stimulation. The progeny of cells which were stimulated with periodate can be restimulated by treatment with either periodate or concanavalin A.
The Mechanism of Basophil Histamine Release Induced by Antigen and by the Calcium Ionophore A23187
Authors: Lichtenstein Lawrence M.
Published: December 2022

The mechanism of human basophil histamine release by the calcium ionophore A23187 has been compared to that induced by the interaction of antigen with cell bound IgE antibody. Ionophore induced histamine release (Ion. H.R.) occurs with the leukocytes of both normal and allergic donors. It is completely calcium dependent; LaCl3 inhibits both Ion. H.R. and antigen induced histamine release (Ag. H.R.) at about 10-7 M. The kinetics of Ion. H. R. suggest that this process has no “desensitization” phase as does Ag. H.R. and the ionophore is fully active on antigen-desensitized cells. Pharmacologic studies indicate that dibutyryl cyclic AMP and agents which increase endogenous cyclic AMP levels do not inhibit Ion. H.R. as they inhibit the early stages of Ag. H.R. Of the agents which affect microtubules, colchicine inhibits and D2O enhances Ion. H.R. in a manner which is qualitatively similar but quantitatively less marked than their effects on Ag. H.R. The metabolic antagonist 2-deoxyglucose inhibits both Ion. H.R. and Ag. H.R. in a similar fashion. Based on these data and the observation that cells pretreated with ionophore show a marked (synergistic) enhancement of Ag. H.R. we conclude that Ion. H.R. has a similar or identical mechanism to the later stages of Ag. H.R. but “short circuits” the cyclic AMP-associated events of Ag. H.R.
Rejection of Mammary Adenocarcinoma Cell Tumors in DBA/2 Mice Immunocompromised by Thymectomy and Treatment with Antithymocyte Serum
Authors: Likhite Vilas V.
Published: December 2022

Syngeneic mammary adenocarcinoma cells were mixed with killed Corynebacterium parvum organisms and were then transplanted subcutaneously into groups of normal and immunocompromised mice. The tumors formed at the site of injection exhibited normal growth for approximately 12 days followed by rapid and lasting rejection in both normal and immunocompromised animals. The control mice were protected against reinjections of 104 to 108 cells of the same line of tumor cells. The immunocompromised mice exhibited protection to reinjection of 104 cells of the tumor line initially employed. Microscopic examination of histologic sections of 10-day-old tumors in immunocompromised mice arising from administration of tumor cells and C. parvum mixtures revealed infiltration of only macrophages whereas those from normal animals exhibited both macro-phages and lymphocytes. The data suggested that primary rejection of these tumors in immunocompromised mice may be the result of macrophage activity together with humoral immunity. These studies also revealed that 59 to 75% of immunocompromised mice exhibit protection against reinjection of the same line of tumor cells if twice-weekly treatment (i.p.) with killed C. parvum was included after inoculation of tumor cells mixed with C. parvum vaccine. These mice, unlike their saline-treated peers, exhibit delayed hypersensitivity reactions which are associated with proliferation of θ antigen-bearing lymphocytes. They also have intact tumor-directed humoral immune reactions.