bluuebunny/crossref_metadata_2025 huggingface dataset:
The Antibody Response to Myelin Basic Protein (BP) in Lewis Rats: The Effect of Time, Dosage of BP, and Dosage of Mycobacterium Butyricum
Authors: Day Eugene D. , Pitts Olin M.
Published: December 2022

Through the use of a sodium sulfate method of radioimmunoassay the influence of variables such as the dosage of myelin basic protein (BP), the dosage of Mycobacterium butyricum, and the presence or absence of Bordetella pertussis was studied with reference to time of appearance and final level of anti-BP antibody in Lewis rats. Animals injected with constant amounts of mycobacteria (250 µg) and variable amounts of BP (2 to 50 µg) in Freund's complete adjuvant showed antibody levels which were proportional to the dosage of BP injected. The appearance of experimental allergic encephalomyelitis (EAE) could not be correlated with antibody level. When doses of BP were held constant (2.5 µg) while doses of injected mycobacteria were varied (5 to 500 µg), animals receiving higher amounts of mycobacteria developed antibody later and at lower levels than animals receiving an intermediate dosage of 50 µg. Parameters found to have very little influence upon antibody levels were the administration of 6.4 × 109 B. pertussis organisms as an ancillary adjuvant and the frequency of animal bleeding. Mycobacteria control injections in complete Freund's adjuvant without BP were shown to induce anti-BP antibody formation above normal, particularly at lower doses, but adsorption studies showed no evidence for BP-mycobacteria cross-reactivity. The hypothesis was raised and discussed that a) a normal animal contains a few active B-cells which produce low levels of anti-BP antibody, that b) this non-proliferative B-cell response is antagonistic to and/or competitive with the cell-mediated EAE-producing T-cell response, that c) the injection of BP antigen and mycobacteria in high enough doses in complete Freund's adjuvant simultaneously neutralizes the normal protective responses and sets the stage for EAE, and that d) the later proliferative B-cell response usually comes too late to be inhibitory.
Molecular Analysis of the Reaction of C9 with EAC1-8: Reaction of C9 with EAC1-8
Authors: Kitamura Hajime , Inai Shinya
Published: December 2022

The hemolytic dose-response curve of either C9gp or C9hu using EAC1-8gp was shown to be concave to the abscissa representing the concentration of C9, but that of either C9gp or C9hu using EAC1-8hu was sigmoidal. When the number of molecules of C9 bound to EAC1-8 was calculated by subtracting the number of residual C9 molecules, which was not bound to the cells after reaction with EAC1-8, from that of added C9, it was found that one molecule of C9 bound to EAC1-8gp could generate about one site, but four or more molecules of bound C9 per cell might be necessary to generate one site on EAC1-8hu. Kinetic studies using EAC1-8hu and C9gp revealed that the generation of E* and lysis of E* proceeded with lag phase and the maximal velocities of these reactions were not proportional to the concentration of C9gp and EAC1-8hu. It was concluded from these results that EAC1-8gp could be lysed by one molecule of either C9gp or C9hu, and that more than one molecule of C9gp or C9hu might be necessary to produce a membrane lesion responsible for the hemolysis of EAC1-8hu.
Cellular Immunity to Leishmania Donovani
Authors: Skov Clare B. , Twohy Donald W.
Published: December 2022

C57BL/6J mice were infected with Leishmania donovani after a series of treatments that were designed to deplete thymus-dependent (T) cell populations. Deprived mice did not acquire immune resistance to the parasite. The capacity to resist infection was restored by either lymph node cells (LNC) or thymocytes, but 20 to 25 times more thymocytes than LNC were required. Reconstitution of immunocompetence by both cell types was inhibited by in vivo treatment with azathioprine or anti-brain associated θ serum. Limiting dilution experiments showed that constant minimal numbers of LNC combined with graded numbers of thymocytes, or vice versa, conferred greater protection than expected from the additive effect of both cell types acting independently. Cultured peritoneal macrophages, harvested from normal and reconstituted mice 44 days after infection, suppressed parasite multiplication. Macrophages from infected deprived mice were unable to prevent parasite multiplication in vitro. These results provide strong evidence that acquired resistance of mice to L. donovani involves T-T cell synergy and relies principally on cellular immunty.
Rat-Mouse Radiation Chimeras: Characterization of an Antibody-Mediated Graft-Vs-Host Reaction
Authors: Kateley John R. , Gengozian Nazareth
Published: December 2022

A modification of the Jerne hemolysis-in-agar technique was used to demonstrate antibody-mediated graft-vs-host (GvH) hemolytic activity in lymphoid tissues of rat-mouse radiation chimeras. Initial foci of hemolytic activity were detected on the 3rd day in spleens of lethally x-irradiated mice infused with rat spleen cells (950 R-RSp chimeras). The peak GvH foci response occurred near the end of the 1st week when 70% of 950 R-RSp chimera spleens examined contained an average of 18 to 21 foci per spleen. GvH hemolytic activity was also detected in mice reconstituted with rat bone marrow (950 R-RBM chimeras); however, only 40% of these chimeras contained splenic foci (nine foci per spleen) on day 8, the peak day of response. Mesenteric lymph nodes from 950 R-RSp chimeras, but not 950 R-RBM chimeras, also contained GvH hemolytic foci. Specificity experiments demonstrated that the GvH hemolytic reaction could be inhibited with rabbit anti-rat IgM serum and host (C3BF1) antigenic cell fragments. Indirect (IgG) hemolytic activity was also observed in spleens of 950 R-RSp chimeras 6 to 10 days after treatment. At least 90% of indirect foci appeared coincident with direct foci. Rat IgM and IgG containing cells beneath hemolytic foci were observed by combining the fluorescent antibody and foci techniques. To determine whether the foci response reflects an in vivo sensitization of donor cells, spleen cell transfer experiments were done. The results indicate that spleen cells from 950 R-RSp chimeras were more effective in accelerating mortality and anemia and induced a higher incidence of Coombs' positive red cells in lethally irradiated isogeneic recipients than spleen cells from 950 R-RBM chimeras; spleen cells from either chimera were more effective in accelerating GvH indices than spleen and marrow cells from normal rats.
Spontaneous and Methylcholanthrene-Enhanced Thyroiditis in Buf Rats
Authors: Silverman David A. , Rose Noel R.
Published: December 2022

The immunization of Buffalo strain (BUF) rats with rat thyroid extract and Bordetella pertussis vaccine without complete Freund's adjuvant resulted in experimental autoimmune thyroiditis (EAT) in 90% of the rats by the 10th day after immunization. The treatment did not elicit thyroiditis in Lewis strain (LEW) rats. This susceptibility to the induction of EAT without the use of complete Freund's adjuvant was inherited by F1 hybrids of BUF and LEW. Neither histamine sensitizing factor from B. pertussus nor Escherichia coli vaccine could substitute for whole pertussis vaccine.
Dissociation of the Immunologic Release of Histamine and Slow Reacting Substance of Anaphylaxis from Human Lung Using Cytochalasins A and B
Authors: Orange Robert P.
Published: December 2022

Cytochalasins A and B at low doses resulted in enhancement of the antigen-induced and anti-IgE-mediated release of histamine from human lung fragments in vitro. Both compounds appeared to effect a dose-dependent inhibition of the antigen-induced release of slow reacting substance of anaphylaxis. Although the effects observed with cytochalasin B were at least partially reversible, the effects of cytochalasin A could not be readily reversed. The cytochalasins appeared to be more effective when present during the antigen-dependent activation phase of mediator release rather than during the calcium-dependent release phase.
An in Vitro Method for Study of Human Lymphocyte Cytotoxicity against Mumps-Virus-Infected Target Cells
Authors: Andersson Torbjörn , Stejskal Vera , Härfast Bengt
Published: December 2022

A chromium release assay was used to study lymphocytemediated cytotoxicity against mumps virus-infected target cells in vitro. Purified lymphocytes from randomly selected donors killed significantly more virus-infected Vero cells than non-infected cells. Lymphocyte-target cell ratios of 50 to 100:1 and incubation period from 16 to 20 hr were optimal for determination of cytotoxicity. The lymphocyte induced chromium release was not obviously correlated with serum mumps hemagglutination-inhibition titers of the effector cell donors. However, the lymphocyte reaction against virus-infected target cells seems to have an immunologic basis. Thus, the more pronounced susceptibility of mumps-infected target cells as compared to non-infected cells was not due to a cytocidal effect of the virus, since spontaneous isotope release from both target cells was the same. Also, cord blood lymphocytes which had exhibited good cytotoxicity when induced either with phytohemagglutinin or with antiserum against target cell antigens were not cytotoxic for mumps-infected Vero cells. Moreover, the lymphocyte reaction against virus-infected target cells could be inhibited by high concentrations of hyperimmune rabbit antimumps sera. On the other hand, lower concentrations of antisera specifically potentiated the lymphocyte-mediated isotope release from mumps-infected target cells.
Characterization of Cytotoxic Effector Cells in the Mouse Mammary Tumor System
Authors: Lane Mary-Ann , Roubinian Jirayr , Slomich Maxwell , Trefts Park , Blair Phyllis B.
Published: December 2022

Cell types involved in the immune response in vitro to MTV-induced BALB/cfC3H mammary tumors have been studied with techniques designed to inactivate or deplete different cytotoxic effector cells from spleen cell populations. The minimal activity of spleen cells from neonatally MTV-infected virgin BALB/cfC3H females is dependent upon the presence of T cells. Spleens from multiparous BALB/cfC3H females have a more active T cell population, and, in addition, a non-T cell population which is active early in the culture period. Responses of spleen cells from multiparous BALB/cfC3H females bearing small tumors are similar to those of tumor-free multiparous females. However, after the tumors grow to large size, the activity of the non-T cell population is no longer detectable. The cytotoxic activity of spleen cells from BALB/c females older than 14 weeks (supposedly not infected with MTV) is not dependent upon the presence of T cells. No activity is shown by spleen cells from BALB/c females younger than 14 weeks. Positive control spleen cells were obtained from C3H female mice who had been immunized to BALB/c histocompatibility antigens by a skin allograft; both a T cell and a non-T cell response were found.
Similar Idiotypic Specificity for the Membrane IgD and IgM of Human B Lymphocytes
Authors: Fu S. M. , Winchester R. J. , Kunkel H. G.
Published: December 2022

Certain cases of chronic lymphocytic leukemia possess monoclonal bands in the serum. Idiotypic antibodies to the isolated IgM protein of one such case demonstrated that the leukemic lymphocytes carried the identical specificity on their lymphocytes. Both the lymphocyte IgM and the IgD possessed this same specificity. This was demonstrated best through the use of rhodamine-conjugated Fab fragments of IgM- and IgD-specific antisera which were both capped by the idiotypic antiserum.
A Radioimmunoassay for Human Antigen-Antibody Complexes in Clinical Material
Authors: Cowdery John S. , Treadwell Perry E. , Fritz Robert B.
Published: December 2022

Soluble antigen-antibody complexes have been detected by a radioimmunoassay with rheumatoid factor as the primary antibody to bind 125I immune complexes or heat aggregated IgG. Bound complexes are precipitated by rabbit anti-human IgM. The binding of the rheumatoid factor to the iodinated altered IgG can be inhibited by preincubation of the rheumatoid factor with human antigen-antibody complexes. The degree to which immune complexes inhibit the binding of rheumatoid factor to heat aggregated IgG has been shown to be dependent upon the antigen-antibody ratio of the complexes, with complexes formed at or near the point of equivalence exhibiting maximum inhibitory activity. Sera from individuals suffering from certain connective tissue disorders were found to inhibit the binding of rheumatoid factor to 125I aggregated IgG; sera from normal individuals did not inhibit this reaction when used at comparable dilutions. This procedure can be used as a relatively simple, reproducible means of detecting immune complexes in clinical specimens. The assay is sufficiently sensitive to detect 125 ng of soluble immune complexes.
Quantitative Determination of Allotypic and Idiotypic Markers with Antisera Coupled to N-Hydroxysuccinimide-Activated Sepharose
Authors: Gottlieb A. Bendix , Seide R. K. , Kindt T. J.
Published: December 2022

N-hydroxysuccinimide activated Sepharose beads (HAS) were used to immobilize allotypic and idiotypic antisera for use in radioimmunoassays. These HAS antisera were compared to antisera solidified with ethylchloroformate (ECF) with respect to their reactivity, specificity, and ability to detect subspecificities of the group a allotypes. The HAS antisera rated favorably in all comparisons with ECF antisera and are far superior with respect to ease of preparation and retention of antibody activity.
The Release of Four Mediators of Immediate Hypersensitivity from Human Leukemic Basophils
Authors: Lewis Robert A. , Goetzl Edward J. , Wasserman Stephen I. , Valone Frank H. , Rubin Robert H. , Austen K. Frank
Published: December 2022

The availability of a patient with basophilic leukemia manifesting 75 to 90% mature basophils permitted the use of a cell concentration sufficient to generate and release mediators upon interaction with a calcium ionophore in quantities adequate for their physicochemical characterization. The mediators were defined in terms of their physicochemical characteristics: slow reacting substance of anaphylaxis (SRS-A) by purification through silicic acid chromatography and inactivation by arylsulfatase; eosinophil chemotactic factor of anaphylaxis (ECF-A) by its gel filtration through Sephadex G-25 and inactivation by subtilisin and not trypsin; and platelet-activating factor (PAF) by its inherent binding to albumin. Both ECF-A and histamine were present in their preformed state, and for histamine it was possible to establish that the concentration per cell was comparable to that of normal human basophils. Dibutyryl cyclic AMP suppressed release of histamine and SRS-A, indicating that their availability was under a control similar to that observed with normal cells subjected to immunologic activation. The demonstration that a suspension of leukemic human basophils contained the preformed mediators, histamine and ECF-A, and generated SRS-A and PAF for release along with histamine and ECF-A, after activation with a calcium ionophore, establishes that a single cell type can serve as a source of the four recognized mediators of immediate-type hypersensitivity.
Phylogeny of Complement Components in Non-Human Primates
Authors: Schur Peter H. , Connelly Anne , Jones T. C.
Published: December 2022

The antigenic properties and functional activities of complement components were analyzed in primates to determine their relative evolutionary development. The sera of eight different sub-human primate species were examined by double diffusion in agar and compared to a pool of human serum with rabbit and goat antisera to human complement components C1q, C1s, C4, C2, C3, C5, C6, C8, C9, properdin, factor B (B), and C1 inhibitor. There are no apparent antigenic differences in complement proteins between man and the apes except for C1q. Old World monkeys are antigenically deficient in C1q, C1s, C9, and variably deficient in C4, C3, and C8. New World monkeys are antigenically deficient in all components (measured) except C5, C6, and properdin factor B. Prosimians are antigenically deficient in all components. Functional analyses of complement components showed similar levels in man and primates, except in Prosimians. There is a dissociation between hemolytic assays and antigenic analyses, suggesting that functional sites may be separate from antigenic sites.
Characterization of Chemotactic Activity Produced in Vivo by a Cell-Mediated Immune Reaction in the Guinea Pig
Authors: Postlethwaite Arnold E. , Snyderman Ralph
Published: December 2022

A system that allows repeated sampling of peritoneal fluid at various time intervals has been adapted to study mechanisms of leukocyte accumulation in vivo. Application of this technique in guinea pigs exhibiting delayed hypersensitivity (DH) to horse radish peroxidase (HRPO) has allowed characterization of some events after i.p. challenge with the sensitizing antigen. Within 24 hr of the administration of HRPO i.p. to such animals there is a significant increase in the number of peritoneal macrophages and in the chemotactic activity (CTX) for macrophages in the sampled peritoneal fluid. At 48 and 72 hr the CTX returns to the pre-challenge level and i.p. macrophages appear to be actively phagocytic.

Molecular sieve chromatograms of concentrated peritoneal fluid obtained 24 hr after i.p. challenge with HRPO and of supernatants derived from immune spleen cells cultured in the presence of HRPO in vitro revealed that the major portion of CTX for homologous macrophages eluted in the region of the 12,500 dalton protein marker. The partially purified CTX obtained from peritoneal fluid and supernatants of spleen cell cultures was heat stable (56°C for 30 min) and was destroyed by trypsin digestion.

These data demonstrate that a chemotactic factor for macrophages, similar to a lymphocyte-derived chemotactic factor (LDCF) obtained in vitro, is present in vivo at the site of a cell-mediated immune reaction. Moreover, these observations demonstrate the feasibility of studying the kinetics of leukocyte accumulation and the production of mediators of inflammation at the site of well defined immunologic reactions in vivo.
Quantitative Comparison of Techniques Used to Measure Complement-Mediated Cytotoxicity of Nucleated Cells
Authors: Detrick-Hooks Barbara , Borsos Tibor , Rapp Herbert J.
Published: December 2022

The cytotoxicity of nucleated cells by specific antibody and complement can be quantitated in vitro by several methods. Trypan blue exclusion, 51Cr release, inhibition of uptake of 3H-thymidine, and inhibition of colony formation are the four assays that we used to quantitatively compare the C-mediated cytotoxicity of Chinese hamster lung (CHL) cells. CHL cells were sensitized with either guinea pig or rabbit anti-CHL antisera, and exposed to guinea pig, human, or rabbit C. We found that the relative activities of the antibodies and the complements were not dependent on the method of measurement of cytotoxicity, i.e., a given pair of antiserum-complement combination had the same antiserum-complement titer, regardless of the assay used to measure cytotoxicity. However, the titers of the different antibody-complement pairs differed from each other.
Increased Immunologic Reactivity between Human Glomerular Basement Membrane and Group A Type 12 Streptococcal Cell Membrane after Carbohydrase Treatment
Authors: Blue William T. , Lange Charles F.
Published: December 2022

Anti-streptococcal cell membrane and anti-human glomerular antisera were evaluated on normal human kidney sections by an indirect immunofluorescent test both before and after absorptions with homologous and heterologous membrane preparations. The observed cross-reactions were potentiated by removal of carbohydrate units from both membrane-types by carbohydrase, a mixture of carbohydrate-cleaving enzymes.
Migration Inhibitory Factor and Type II Interferon in the Circulation of Mice Sensitized with Mycobacterial Components
Authors: Salvin S. B. , Ribi E. , Granger D. L. , Youngner J. S.
Published: December 2022

Migration inhibitory factor (MIF) and Type II interferon were released into the circulation after mice had been sensitized i.v. with a bacillus of Calmette and Guerin (BCG) cell-wall-in-oil vaccine and then challenged four weeks later with 50 mg old tuberculin. At least two components of BCG, protein (PPD) and lipid (P3), have been identified which are essential for the type of sensitization in which mediators are released after appropriate challenge. The route of sensitization had a marked effect on the development of delayed footpad reactions, release of lymphokines, and resistance to infection with virulent tubercle bacilli. Sensitization by the subcutaneous route tended to induce more pronounced delayed footpad reactions, whereas the i.v. route of sensitization was associated with maximum release of mediators. A close correlation existed between the conditions of sensitization that resulted in maximum production of lymphokines (MIF and Type II interferon) and those that caused protection against aerosol challenge with a virulent strain of Mycobacterium tuberculosis.
Immunofluorescent Studies on Antibodies Directed to a Buried Membrane Structure Present in Lymphocytes and Erythrocytes
Authors: Winchester R. J. , Fu Shu Man , Winfield J. B. , Kunkel H. G.
Published: December 2022

Brief digestion of human peripheral blood lymphocytes by vibrio cholera neuraminidase (VCN) revealed hidden components of the membrane. Autologous human serums contained antibodies directed to these components that were readily demonstrated by immunofluorescence. Antibodies of similar specificity were found in all normal serums. The antibodies were principally of the IgM variety with lesser amounts of the IgG class present. They were equally active at 4°C and 37°C. The VCN revealed membrane determinants were present in normal B and T lymphocytes, monocytes, lymphocytes of patients with chronic lymphatic leukemia and cells of lymphoid lines. The newly revealed determinants slowly disappeared upon culture of the lymphocytes.

These hidden components were similarly demonstrated in erythrocyte membranes and represent the T antigen long known for the red blood cells. Absorption by either VCN treated autologous lymphocytes or erythrocytes removed all of the antibodies capable of reacting with both cell types. Absorption by VCN digested isologous lymphocytes removed all reactivity with autologous lymphocytes.
A Lectin Model for Unresponsiveness
Authors: Chauvenet Allen R. , Scott David W.
Published: December 2022

A mitogen model for unresponsiveness has been studied. In 72-hr cultures the lowest supraoptimal dose of Concanavalin (Con A) showing no stimulation of rat thymocytes above background was 160 µg/ml. This dose was also found to inhibit substantially (50%) the high spontaneous DNA synthesis by thymocytes during the first 24 hr of culture. This effect was most significant 12 to 24 hr after addition of high-dose Con A. It was not necessary for the cells to remain continuously with high-dose Con A, since washing out unbound Con A after 1 hr still led to reduced 3H-thymidine incorporation at 24 or 72 hr. Furthermore, high-dose Con A treatment suppressed DNA synthesis when added at 48 hr to cultures optimally stimulated with Con A. Interaction with high-dose Con A appears to inhibit the ability of cells to initiate new rounds of DNA synthesis without blocking ongoing DNA replication. Removal of Con A with α-methyl-D-mannopyranoside (mannose) led to only partial reversal of these effects after 1 hr although “unresponsiveness” was reversible at 15 min. Suppression was not due to cytotoxic or agglutinating effects of the high-dose Con A. Two × 106 thymocytes exposed to 160 µg of Con A for 1 hr bound 1.7 µg, 86% of which was elutable with mannose. There was no reduction in the ability of high-dose treated cells to bind a subsequent optimal dose of Con A. Since Con A binds to essentially all thymocytes, this system is useful in following early events in the induction of lectin “unresponsiveness” as a model for immunologic tolerance.
Canine Radiation Chimeras: An Attempt to Demonstrate Serum Blocking Factors by an in Vivo Approach
Authors: Schroeder Maria-Louise , Storb Rainer , Graham Theodore C. , Weiden Paul L.
Published: December 2022

An attempt was made to demonstrate in vivo the presence of blocking factors in the serum of canine chimeras with grafts of chimeric skin onto the marrow donor as an indicator system. Even very large amounts of chimeric serum (one-fourth of the plasma volume infused daily from day 2 before skin grafting until rejection) failed to modify first and third set skin graft rejection patterns. Thus, serum blocking factors could not be demonstrated by this in vivo approach.
Lack of B Lymphocyte Depletion from Murine Spleen Cell Populations by a Human γ-Globulin, Anti-Human γ-Globulin Column System
Authors: Karpf Michael , Gelfand Michael C. , Handwerger Barry S. , Schwartz Ronald H.
Published: December 2022

It has been reported previously that enriched populations of mouse thymus-dependent (T) lymphocytes could be prepared by the use of a column procedure that is believed to remove selectively cells with receptors for antigen-antibody complexes. In our hands, this procedure does not selectively remove B lymphocytes but rather masks the surface immunoglobulin. Analysis of the column-passed cells revealed an apparent decrease in the percentage of immunoglobulin-bearing cells but little decrease in either the percentage of complement receptor lymphocytes or the mitogenic response to lipopolysaccharide. Furthermore, rabbit immunoglobulin could be detected on the surface of 35 to 45% of the cells emerging from the columns prepared with rabbit anti-human γ-globulin (HGG). After overnight incubation, 30 to 40% of the emerging cells reexpressed mouse immunoglobulin on their surfaces. Adsorption of the antisera used to make the columns with mouse γ-globulin (MGG) diminished the depleting capacity of the columns. We conclude from these observations that antibodies in the anti-HGG sera, cross-reactive with mouse immunoglobulin, account for the specious depletion of B lymphocytes observed when cells are passed over these antigen-antibody complex columns.
Preliminary Evidence for Cell-Mediated Immunity in Cross-Protection Among Group A Arboviruses
Authors: Peck Richard D. , Brown Arthur , Wust Carl J.
Published: December 2022

Unfractionated and T cell-enriched populations from spleens of mice immunized with Sinbdis virus confer cross-protection against Semliki forest virus challenge in recipient mice. Both Group A viruses are serologically related, but not by tests for cross-neutralizing antibody. The protection in mice persisted for three consecutive cell passages in uninfected mice. The results suggest that cell-mediated immunity accounts for cross-protection.
Regulation of Antibody Response in Vitro
Authors: Kishimoto Tadamitsu , Ishizaka Kimishige
Published: December 2022

Attempts were made to induce antibody response of hapten-primed cells by stimulation with anti-immunoglobulin (Ig) antibody and cellfree supernatant (CFS) which contained nonspecific enhancing factor. Mesenteric lymph node cells were obtained from rabbits which were primed with dinitrophenylated ragweed antigen (DNP-Rag), and the primed cells were incubated in vitro with either anti-rabbit γ-chain or anti-Fab antibody for 24 hr. After washing, the cells were cultured in CFS which was obtained from the culture of DNP-Ascaris (DNP-Asc)-primed lymph node cells with free carrier (Asc). The results showed that hapten-primed cells were triggered for IgG anti-DNP antibody response by the two reagents, i.e., anti-Ig antibody and CFS, both of which did not share any antigenic specificity with the priming antigen (DNP-Rag). Neither anti-Ig alone nor CFS alone induced anti-hapten antibody response. Anti-Ig and CFS triggered not only hapten-primed B cells but also the other IgG-bearing (B) cells for IgG synthesis. Lymph node cells of unprimed animals were activated for IgG synthesis by the stimulation with anti-Ig followed by culture in CFS. Evidence was obtained that anti-Ig antibody has to be divalent for the activation of B cells. Stimulation of DNP-primed cells with the F(ab′)2 fragment of the antibody and CFS-induced anti-hapten antibody response, whereas the Fab′ fragment of the anti-Ig plus CFS failed to do so. The results suggested that bridging of cell-surface Ig receptors by multivalent ligand is the initial step of B cell activation. Another evidence for the activation of hapten-primed B cells by anti-Ig was obtained by supplemental immunization of DNP-Asc-primed animals with the Fc fragment of goat IgG. Stimulation of mesenteric lymph node cells from these animals with goat anti-rabbit-Ig alone (no CFS) resulted in the formation of anti-DNP IgG antibody. The results indicated that lymphocytes primed for goat IgG collaborated with DNP-primed IgG-B cells when the lymph node cells were stimulated with goat anti-Ig.
The Location of Regions in Guinea Pig and Bovine Myelin Basic Proteins Which Induce Experimental Allergic Encephalomyelitis in Lewis Rats
Authors: Martenson Russell E. , Levine Seymour , Sowinski Richard
Published: December 2022

Myelin basic proteins and peptides derived from them by limited cleavage with pepsin were tested for their ability to induce experimental allergic encephalomyelitis (EAE) in Lewis rats. The encephalitogenicity of the weakly active bovine protein was found to be associated with both halves of the molecule, peptides (1–88) and (89–169). Of the four smaller derivatives of peptide (1–88), peptides (1–36), (43–88), (1–42), and (37–88), only the last two were active. This demonstrated that the overlap region consisting of residues 37–42 (sequence Asp-Ser-Leu-Gly-Arg-Phe) constitutes an encephalitogenic determinant. Of the two smaller derivatives of peptide (89–169), peptides (111–169) and (89–152), only the last was active. This indicated that the second encephalitogenic determinant begins between residues 88 and 111 and ends before residue 153. This region contains the sequence Leu-Ser-Leu-Ser-Arg-Phe (residues 108–113), which is strikingly similar to that of the first encephalitogenic determinant. Studies involving the extremely encephalitogenic guinea pig protein demonstrated that virtually all of the activity was recovered in the peptides corresponding to bovine peptides (37–88) and (43–88). These peptides, but not those comprising the remainder of the protein, were active in inhibiting the passive transfer of EAE with lymph node cells from donors immunized with guinea pig spinal cord.
Allergic Encephalomyelitis in the Reputedly Resistant Brown Norway Strain of Rats
Authors: Levine Seymour , Sowinski Richard
Published: December 2022

Brown Norway (BN) rats are much less susceptible to experimental allergic encephalomyelitis (EAE) than Lewis rats. Nevertheless, BN rats developed severe EAE, even paralysis, when immunized with rat spinal cord and carbonyl iron adjuvant. Complete Freund's adjuvant (CFA) was much less effective. The use of both CFA and pertussis vaccine with rat cord was moderately, but not consistently, effective. Guinea pig spinal cord was weakly encephalitogenic to BN rats with all adjuvant combinations. We were not able to produce EAE in BN rats with purified myelin basic protein from either rat or guinea pig. Inoculations directly into lymph nodes or into the blood stream proved that the low susceptibility of BN rats was not due to lack of absorption from the site of inoculation, but may be related to peculiarities of processing antigen in draining lymph nodes. The severity of EAE in F1 hybrids was intermediate between the BN and Lewis parental strains when tested with an immunizing procedure of appropriate strength. The fact that F1 hybrids were less reactive than Lewis mandates modification of the theory that susceptibility to EAE is inherited through a single autosomal dominant gene.
Characterization of Immune Response and Mixed Lymphocyte Reactions in Selected Intra-H-2 Recombinant Strains
Authors: Dorf M. E. , Plate J. M. D. , Stimpfling J. H. , Benacerraf B.
Published: December 2022

The genes controlling the immune response to the random linear terpolymers GAT and GLpro have been mapped in the Ir-1A or Ir-1B subregions with five lines of H-2a/H-2a recombinant mice. The I region also contains a third subregion, I-C, which codes for a lymphocyte-activating determinant, Lad2, controlling mixed lymphocyte reactivity. The mixed lymphocyte responses associated with I-C region disparity are compared with the stimulation noted with other H-2 region differences. Unidirectional lymphocyte activitation was associated with the Lad2 determinants in selected combinations.
Reaginic Antibody Formation in the Mouse
Authors: Okudaira Hirokazu , Ishizaka Kimishige
Published: December 2022

DBA/1 mice were primed with dinitrophenyl-keyhole lympet hemocyanin (DNP-KLH) included in aluminum hydroxide gel, and the adoptive anti-DNP IgE antibody response of their spleen cells was studied by transfer into irradiated syngeneic mice. As expected, depletion of T cells by anti-ϑ antiserum and complement abolished the response of the spleen cells to homologous antigen. If the same T-depleted spleen cells were cultured with DNP-KLH for 24 hr and then transferred into irradiated mice, anti-DNP IgE antibody response was obtained. It was also found that DNP-KLH-primed spleen cells were triggered for IgE antibody response if they were cultured for 24 to 48 hr with DNP heterologous carrier conjugate, such as DNP-bovine γ-globulin or a copolymer of D-tyrosine, glutamine, and lysine. Transfer of the cells cultured with the antigen into irradiated recipients resulted in the formation of anti-DNP IgE antibody. The DNP-KLH-primed cells, cultured for 24 hr in the absence of antigen and then treated with DNP-BGG at 0°C, also gave an adoptive IgE antibody response. if the same DNP-KLH-primed cells were treated with the DNP heterologous carrier conjugate at 0°C or injected into irradiated recipients together with the antigen, no IgE antibody response was obtained. The results indicated that T cell dependency of the IgE antibody response is diminished by culture of DNP-KLH-primed cells for 24 to 48 hr.
Neural Antigens and Induction of Myelination Inhibition Factor
Authors: Seil Fredrick J. , Kies Marian W. , Bacon Martha
Published: December 2022

Rabbits were sensitized or immunized with a variety of central nervous system antigens, including bovine spinal cord, bovine, monkey, human, guinea pig, rabbit, and rat S myelin basic proteins, and a polypeptide derived from guinea pig basic protein. The animals were observed for development of experimental allergic encephalomyelitis, and their sera were collected at varying intervals after inoculation and evaluated for presence of precipitating anti-basic protein antibody and for their ability to inhibit myelin formation in cerebellar tissue cultures. The resulting complete dissociation between development of experimental allergic encephalomyelitis, the presence of antibasic protein antibody and the occurrence of myelination inhibition factor suggests that myelination inhibition factor is not involved in the pathogenesis of experimental allergic encephalomyelitis, and argues against a role for anti-basic protein antibody as an antimyelin factor in vitro.
Heavy Chain-Producing Variants of a Mouse Myeloma Cell Line
Authors: Morrison Sherie L. , Scharff Matthew D.
Published: December 2022

Eleven variants producing only heavy (H) chains have been isolated from the P3 cell line after mutagenesis with either ICR-191 or nitrosoguanidine. The H chains produced by these variants are similar both in molecular weight and by peptide maps with the parental H chain. H chains dimerize intreacellularly, are not secreted but persist as stable cytoplasmic proteins. No evidence for the continued synthesis of light (L) chains or L chain fragments has been found in these variants. These studies show that, in the P3 cell line, the synthesis of H chains can continue in the absence of detectable L chain synthesis.
Lymphoid Cell Adenylate Cyclase Activity after X-Irradiation and Cortisone Treatment
Authors: Kemp Robert G. , Duquesnoy René J.
Published: December 2022

Cortisone injection and x-irradiation of normal mice lead to extensive destruction of thymus and spleen lymphocytes. The lymphocytes resistant to these treatments have basal adenylate cyclase activities that are the same as or slightly lower than the cells of untreated animals. On the other hand, the ability of the cyclase of the cortisone-resistant and radioresistant lymphocytes to respond to stimulation by epinephrine is greatly diminished. The results provide an explanation for the previously observed lack of epinephrine sensitivity of the cyclase of lymphoid cells of immunodeficient pituitary dwarf mice and suggest that the depressed lymphoid cell population of the dwarf is the result of an inability to recover from stress-related lymphoid involution. It is also suggested that the more mature population of lymphoid cells of normal mice is less responsive to hormonal influences.
The Production of Anti-Guinea Pig Lymphokine Antibody
Authors: Yoshida Takeshi , Bigazzi Pierluigi E. , Cohen Stanley
Published: December 2022

Rabbit antiserum to lymphokine-containing supernatant fluids has been prepared by means of a two-stage immunization procedure. Migration inhibiting factor (MIF)-containing and control supernatants were obtained from guinea pig lymphocyte cultures. Both were fractionated on Sephadex G-100. Fractions of control supernatants corresponding to active fractions from the MIF-rich material were used to immunize rabbits. The anti-“control” antibody so obtained was conjugated to agarose beads to prepare an immunoadsorbent column. The MIF-rich Sephadex fractions were passed through this column to remove materials corresponding to those found in control supernatants and which could react with the anti-control antibody. The eluted MIF-rich fraction was concentrated and used to immunize a second group of rabbits. The anti-“lymphokine” antiserum so obtained was studied for its effect in vitro and in vivo. It was found capable of suppressing the activity of MIF, macrophage chemotactic factor, and skin reactive factor. It could also suppress delayed hypersensitivity skin reactions in actively immunized guinea pigs.
Synergistic Effects on DNA Synthesis of Phytohemagglutinin or Concanavalin A and Lipopolysaccharide in Human Peripheral Blood Lymphocytes
Authors: Schmidtke Jon R. , Najarian John S.
Published: December 2022

Mixtures of phytohemagglutinin (PHA) or Concanavalin A (Con A) and lipopolysaccharide (LPS) with human peripheral blood lymphocytes (PBL) resulted in a synergistic effect on DNA synthesis as measured by increased 3H-thymidine uptake. These data were compared to the mitogenic response of human PBL to PHA, Con A, or LPS alone. Various combinations of PHA and Con A usually suppressed DNA synthesis in human PBL. Titration experiments revealed that increasing amounts of LPS, in the presence of a constant amount of Con A, resulted in corresponding increases in the synergistic response of human PBL. Conversely, increasing amounts of Con A, in the presence of a constant amount of LPS did not lead to increases in the amount of synergism. The results are discussed in terms of the participation of additional cells in the synergistic response of human PBL.
Antibody-Dependent Cytolysis of Chicken Erythrocytes by an in Vitro-Established Line of Mouse Peritoneal Macrophages
Authors: Walker William S. , Demus Annmarie
Published: December 2022

An in vitro-established line of mouse peritoneal macrophages (IC-21) was tested for its ability to mediate the cytolysis of 51chromiun-labeled chicken erythrocytes. In the presence of specific antibody, but independently of complement, the macrophages phagocytized and lysed labeled erythrocytes. The phagocytic process proved to be functionally distinct from the cytolytic reaction as demonstrated by enhanced cytolysis in the presence of iodoacetate, an inhibitor of phagocytosis.

This cell line, because of its effector activity in antibody-dependent cell-mediated immune reactions, will be useful in characterizing the mechanism(s) involved in macrophage-mediated cytolysis.
Immunologic Properties of Bacterial Lipopolysaccharide (LPS): Correlation between the Mitogenic, Adjuvant, and Immunogenic Activities
Authors: Skidmore Barry J. , Chiller Jacques M. , Morrison David C. , Weigle William O.
Published: December 2022

Bacterial lipopolysaccharide (LPS) was demonstrated to have the capacity in mice to enhance the response to soluble bovine serum albumin (BSA) and to interfere with the induction of tolerance to human γ-globulin (HGG). These adjuvant activities were shown to occur under conditions in which LPS could also function as a B cell mitogen. This positive correlation was established by utilizing two experimental situations in which LPS was non-mitogenic for spleen cells. Thus, on the one hand, it was found that LPS did not function as an adjuvant in C3H/HeJ mice, a unique strain whose spleen cells were also unresponsive to LPS-induced mitogenesis. On the other hand, in strains which did respond to LPS mitogenically, LPS failed to function as an adjuvant when it was chemically altered to reduce its in vitro mitogenic activity. A correlation was also observed between mitogenesis and the capacity of LPS to function as a specific immunogen in mice. In contrast to the sustained and prolonged plaque-forming cell response that was observed in mice whose spleen cells were also responsive to LPS-induced mitogenesis, the response was relatively transient in the C3H/HeJ strain. These results are discussed in view of the possible in vivo modes of action of LPS.
Effect of Polycations and Polyanions on Behavior of Sheep Red Blood Cell Rosettes of Human Lymphocytes
Authors: Yu David Tak Yan , Pearson Carl M.
Published: December 2022

The role of cell surface charges in the behavior of sheep red blood cells (SRBC) rosettes with human lymphocytes was investigated by using various polycations and polyanions. Polycations (DEAE-dextran, polybrene, poly-l-lysine, spermadine) at appropriate concentrations (1) invariably increased the proportions of early rosettes. This was so irrespective of whether it was the SRBC or the lymphocytes that were being treated with the reagents. (2) The rosettes so formed also consisted of a greater number of red blood cells per rosette (3). On incubating the rosettes at 37°C, they dissociated by capping. The rates of dissociation and capping were retarded by polycation treatment. (4) The resistance to rosette disruption was increased. These properties were very similar to those observed on cells treated with neuraminidase. Treatment of cells with the polyanions heparin and dextran sulfate induced the opposite effects: reduced the number of rosettes and increased the rate of dissociation, and decreased the resistance of rosettes to disruption. It was concluded that alterations of cell surface charges could drastically influence SRBC-lymphocyte interactions. To investigate whether rosette inhibitors could do so by altering cell surface charges, cell were treated with anti-lymphocytic sera, 2,4-dinitrophenol and sodium azide. All three agents inhibited rosette formation. Whereas DEAE-dextran could partially reverse the inhibition by the first two agents, it failed to do so with the third. This suggested that the first two may have inhibited rosette formation by altering the surface charges.
Suppression of Reaginic Antibody Formation
Authors: Lee Weng Y. , Sehon Alec H.
Published: December 2022

By the use of the adoptive transfer system in syngeneic mice it was demonstrated that: i) spleen cells of animals tolerized by the i.v. injection of DNP8-MγG 1 month before adoptive transfer into x-irradiated recipients remained unresponsive when further challenged with the sensitizing antigen (DNP2.3-OA administered i.p. with Al (OH)3); by contrast, a significant response (typical of a secondary response) was obtained in recipients of cells from mice which had received only the sensitized antigen; ii) the anti-hapten IgE secondary response of primed cells was not affected by the presence of cells of tolerized mice when these two types of cells were administered and challenged together in x-irradiated recipients; iii) the transfer of spleen cells or serum of mice, tolerized 20 days before harvesting, into intact syngeneic recipients did not interfere with the normal development of the anti-DNP IgE response of the latter; iv) the anti-hapten responses of tolerized animals receiving spleen cells from normal or primed mice together with the sensitizing antigen resulted, respectively, in a primary or secondary response; (v) the unresponsive state of tolerized cells, achieved by immunosuppression of the original donors with respect to the haptenic determinant, was maintained even after two serial exposures of the cells to the immunizing antigen in two consecutive adoptive cell transfers into x-irradiated recipients. These findings provide a strong basis for the interpretation that the hapten-specific tolerance induced in mice by treatment with DNP8-MγG involved the elimination or inactivation of hapten-specific IgE-producing cells or the blockade of the receptors of these cells.
Reticulum Cell Sarcoma: An Effector Cell in Antibody-Dependent Cell-Mediated Immunity
Authors: Ralph Peter , Prichard John , Cohn Melvin
Published: December 2022

A transplantable, murine reticulum cell sarcoma is described which exhibits the cytologic, adherence, and phagocytic properties of macrophages. It forms specific rosettes with erythrocytes in the presence of the corresponding anti-serum. The ascites cells mediate antibody-dependent cellular immunity as assayed by release of radioactivity from 51Cr-labeled erythrocytes. The contribution of contaminating host cells in the cytotoxic reaction was ruled out by growing the tumor in F1 mice and removing the host cells by anti-H2 serum and complement. The tumor cells have receptors for IgG2a and IgG2b immunoglobulins. The availability of a pure population of effector cells in the immune system allows study of the biochemical processes pursuant to lysis of foreign cells.
Constant-Region Cysteine-Containing Peptides of b4 and b9 Rabbit κ-Chains Isolated by a New Diagonal Mapping Procedure
Authors: Goodfliesh Robert M.
Published: December 2022

A diagonal mapping technique for the selective isolation of cysteine-containing peptides was introduced about 10 years ago by Brown and Hartley (1, 2). Peptides are first separated by electrophoresis in one dimension on filter paper. The paper is then dried and exposed to performic acid vapors which cleave and oxidize cystine residues into two cysteic acid residues. The electrophoretic mobility of cysteic acid-containing peptides usually changes because the peptides are smaller and cysteic acid residues are negatively charged. A second electrophoresis is run at right angles to the first dimension. Peptides which do not contain cysteine have the same mobility in both dimensions and lie on a 45° diagonal. Cysteine-containing peptides “map” off the diagonal. I have developed a new version of this technique which is faster and more specific for cysteine-containing peptides. Proteins are first completely sulfitolyzed (3) and then enzymatically digested; cysteine and cystine residues are thus present as negatively charged S-sulfo cysteinyl derivatives (—S—SO3-).