bluuebunny/crossref_metadata_2025 huggingface dataset:
Endogenous Synthesis of Membrane b5 by Lymphocytes from Rabbits Recovering from b5 Allotype Suppression
Authors: Harrison Michael R. , Jones Patricia P. , Mage Rose G.
Published: December 2022

In the rabbit, production of b-locus allotypic determinants present on kappa-type light chains can be suppressed by perinatal exposure to anti-b5 antisera (allotype suppression) (1, 2). We have previously demonstrated that b5-bearing lymphocytes are deleted in totally suppressed rabbits with no circulating b5, and that in spontaneous escape from suppression the appearance of b5-bearing lymphocytes immediately precedes the appearance of detectable serum b5 (3).

However, during recovery from allotype suppression, the proportion of peripheral blood lymphocytes with membrane b5 approaches normal levels, while circulating b5 levels remain chronically and disproportionately depressed (3). Because there is this unusual discrepancy between the number of b5-bearing lymphocytes and the amount of circulating b5 in rabbits recovering from allotype suppression, and because passive adsorption of circulating b5 molecules to lymphocytes not themselves capable of synthesizing b5 could account for the inappropriately large number of b5-bearing cells, we have investigated these lymphocytes for their ability to synthesize actively their own membrane b5 Ig after enzymatic stripping of the cell surface.
Histoincompatibility Recognition Reaction by Irradiated or Mitomycin-Treated Parental Cells against Hybrid Spleen Cells Produces Hybrid Cell Proliferation and Illusion of Reaction of Hybrid against Parent
Authors: Kennedy James C. , Ekpaha-Mensah J. Ampoe
Published: December 2022

The addition of heavily irradiated or mitomycin-treated spleen cells to cultures containing syngeneic spleen cells and foreign erythrocytes may stimulate to some extent the hemolytic plaque-forming cell responses of these cultures, but the addition of semi-allogeneic irradiated or mitomycin-treated spleen cells may cause much greater stimulation. This semi-allogeneic stimulation is completely reciprocal; the plaque-forming cell responses of F1 hybrid spleen cell cultures often are stimulated as much after the addition of non-proliferating parental cells as are the responses of parental spleen cell cultures after the addition of non-proliferating F1 hybrid cells. The possibility that an in vitro manifestation of hybrid resistance might be involved was ruled out. It was found that irradiated or mitomycin-treated cells, although unable to proliferate, were still able to initiate a recognition reaction against histoincompatible cells, releasing in the process a soluble factor which stimulated proliferation of cells of the antibody-forming series. The significance of this finding for “one-way” mixed lymphocyte reactions is discussed.
Fractionation of Antibodies to the Pneumococcal Polysaccharides by Affinity Chromatography
Authors: Cheng William C. , Fraser Kevin J. , Haber Edgar
Published: December 2022

Antibodies to pneumococcal polysaccharides type III and VIII were fractionated by use of cross-reacting immunoadsorbents and gradient elution with cellobiose and NaCl. The polysaccharides were first derivatized with p-nitrobenzyl bromide. After reduction of the nitro group, they were linked to bovine γ-globulin by diazotization and coupling. The protein-polysaccharide complex was then linked to activated sepharose to form an immunoadsorbent. In other experiments, immunoadsorbents were synthesized from polysaccharides which had first been either subject to aminoethylation or to amidation. In some instances, partial acid hydrolysis was carried out on the completed immunoadsorbent. These adsorbents had a high capacity for antibody. By use of several different immunoadsorbents, as well as varying elution programs with cellobiose and NaCl, complex antibody mixtures could be resolved with the isolation of components of unique electrophoretic mobility. Light chains isolated from these preparations were shown to have a single amino acid sequence at the N-terminus. It is apparent that many antibodies to differing determinants on the relatively simple polysaccharide antigens may be elicited and separated on the basis of these properties.
Studies of Biologic and Serologic Activities of Rabbit-IgG Antibody Depleted of Carbohydrate Residues
Authors: Williams Ralph C. , Osterland C. Kirk , Margherita S. , Tokuda Sei , Messner Ronald P.
Published: December 2022

Rabbit IgG preparations containing antibody activity against individual strains of Staphylococcus aureus, Escherichia coli, group A β-hemolytic streptococci, and pneumococci were digested with Diplococcus pneumoniae endoglycosidase enzyme with removal of more than half of Ig-associated carbohydrate. In addition specifically purified rabbit anti-bovine serum albumin (BSA) IgG was similarly treated. After carbohydrate removal no appreciable change in opsonic, complement fixing, or agglutinating activity was noted in anti-staphylococcal, anti-E. coli, or anti-streptococcal IgG preparations, whereas complete loss of both complement fixing and opsonic activity occurred with rabbit IgG containing anti-pneumococcal antibody activity. No change in passive cutaneous anaphylactic reactions was noted after carbohydrate depletion of immunoabsorbent isolated rabbit anti-BSA IgG antibody. These findings indicate heterogeneous influences of carbohydrate residues on biologic activities of various antibody populations.
The Role of the Thymus during the Induction of Tolerance of a Thymus-Dependent Antigen
Authors: Phillips-Quagliata Julia M. , Bensinger Donald O. , Quagliata Franco
Published: December 2022

The effect of neonatal thymectomy on the induction of tolerance of a thymus-dependent antigen was investigated. It was found that some thymectomized (Tx) rats failed to become tolerant after injection of sufficient bovine serum albumin (BSA) to make sham-thymectomized (STx) rats tolerant. This suggested that in the absence of the thymus, thymus-derived (T) cells already in the periphery before thymectomy, as well as bone marrow-derived (B) cells, failed to become tolerant. This hypothesis was tested by injecting mature T cells in the form of non-adherent, adult, syngeneic peripheral blood lymphocytes (PBL) during the period of tolerance induction. This increased the incidence of high responders to challenge in the Tx, BSA-injected (Tx-BSA) group while tolerance was again established in the STx-BSA group. Thymocytes did not substitute for the thymus in facilitating tolerance induction in PBL although thymocytes themselves appeared to become tolerant. Tx, saline-injected (Tx-Sal) recipients of mixtures of PBL and thymocytes made considerably higher responses to BSA than STx-Sal recipients of the same populations. These results suggest that thymocytes and PBL have different requirements for tolerance induction; the thymus itself or a sub-population of T cells, not adequately represented in PBL or thymocytes, exerts a suppressive effect both in the normal immune response and during tolerance induction.
Migration Inhibition of Rat Peritoneal Cells Induced by Streptococcal Mucopeptides. Characteristics of the Reaction and Properties of the Mucopeptide Preparations
Authors: Heymer Berno , Bültmann Burkhard , Schachenmayr Walter , Spanel Reinhard , Haferkamp Otto , Schmidt Willard C.
Published: December 2022

Preparations of mucopeptide were solubilized from formamide-extracted residues of groups A and B streptococcal cell walls by sonication and purified by column chromatography on Sephadex. Other preparations were made by digestion of particulate mucopeptide with lysozyme and Streptomyces albus muramidase. Analysis of these materials showed chemical similarity and a range of molecular size estimated to be greater than 20 × 106 to less than 5 × 103. The larger components reacted serologically as precipitinogens and the smaller as haptenic inhibitors. Mucopeptide preparations of larger molecular size produced migration inhibition reactions (MIR) with peritoneal cells from non-sensitized rats and guinea pigs. In general, MIR and dermonecrotic reactions decreased with decreasing molecular size. Anti-mucopeptide antibody only slightly reduced mucopeptide-induced MIR. Isolation of a migration inhibition factor by Sephadex chromatography from supernatants of cell cultures incubated with mucopeptide was unsuccessful. Stimulation of cells with Bayol F was required to produce cells susceptible to MIR. Under the experimental conditions for demonstration of MIR, mucopeptide did not directly cause lysis or reduce viability of the peritoneal cells used in the test system.
Sequence Variations in b9 Light Chains as Potential V Region Genetic Markers
Authors: Thunberg Allen L. , Lackland Henry , Kindt Thomas J.
Published: December 2022

Amino terminal sequence analyses of b9 light chains from rabbit antibodies to streptococcal carbohydrate have revealed the presence of glutamic acid at position N16 of light chains of the VκIII length. Glutamic acid has not previously been reported at position N16 of any light chain. Quantitative analysis of N-terminal residues of light chains from non-immune b9 IgG samples suggests that glutamic acid is present as a minor component at position N16 in b9 light chains of the V III length. Length determinations for quantitation of subgroups of b9 light chains were carried out by quantitative analysis on the sequencer and verified by determining the yield of PTH-2-14C-SCMC at each step. This refinement is possible because of the invariant cysteine at position N23 found in all light chains thus far sequenced.

Sequence variations such as Glu at position N16 within light chains of a given subgroup can provide genetic markers for following the inheritance of the variable region of rabbit antibody light chains.
Differential Effect of Neuraminidase on the Immunogenicity of Viral Associated and Private Antigens of Mammary Carcinomas
Authors: Simmons Richard L. , Rios Angelyn
Published: December 2022

Firmly established transplantable C3H/HeJ mammary carcinomas can be inhibited by host challenge with Vibrio cholerae neuraminidase (VCN) treated tumor cells. The effect is totally immunospecific, even VCN-treated tumors bearing shared mammary tumor virus (MTV) antigen cannot induce the regression. Thus, VCN is capable of increasing the immunogenicity of the private, unique-unshared tumor antigens on mammary carcinomas; VCN is incapable of increasing the immunogenicity of the shared MTV associated tumor antigen.
Evaluation of Cell-Mediated Cytotoxic Reactivity Against Tumor Associated Antigens with 125I-Iododeoxyuridine Labeled Target Cells
Authors: Oldham Robert K. , Herberman Ronald B.
Published: December 2022

An in vitro lymphocyte cytotoxicity tissue culture assay utilizing 125I-iododeoxy-uridine (125IUdR) labeled target cells is described. Because of the labeling characteristics of 125IUdR this isotope appears to offer advantages over previously used labels. Toxicity of the isotope is a factor to be evaluated with each cell line. The method of calculation of cytotoxicity lends some insight into the spontaneous cell death rate of tissue culture systems as used for cytotoxicity. This assay is of value in detecting tumor-associated antigens in both animal and human tumor systems.
The Immune Response of Inbred Strains of Mice to a “High-Conjugation” and a “Low-Conjugation” 2,4 Dinitrophenyl-Glutamic Acid57-Lysine38-Alanine5 (DNP-GLA5) Copolymer
Authors: Warner Carol M. , Maurer Paul H. , Merryman Carmen F. , Schmerr Mary Jo F.
Published: December 2022

The response of inbred mouse strains C57BL/6J and C3H/HeJ to a “high-conjugation” (75%) and a “low-conjugation” (τ;0, ≤10%) 2,4 dinitrophenyl-poly (glu57lys38ala5) copolymer, DNP-GLA5, was tested. The C57BL/6J mice were non-responders to the DNP group at doses from 0.07 µg to 82 µg of each antigen. The C3H/HeJ mice were responders to the DNP group. There was no qualitative difference between the two antigens in eliciting both primary and secondary antibody responses. Although there was a large range in response to the DNP group by the C3H/HeJ mice, there was a distinct gap between non-responder and responder mice. The response to the DNP group, when coupled to GLA5, in these two strains of mice, parallels the response to the carrier, GLA5.
Production and Properties of Migration Inhibitory Factor and Interferon in the Circulation of Mice with Delayed Hypersensitivity
Authors: Youngner J. S. , Salvin S. B.
Published: December 2022

Further evidence is presented of the coordinate production and similar properties of two mediators, migration inhibitory factor (MIF) and interferon, in the circulation of BCG-infected mice inoculated with specific antigen (Old Tuberculin, OT). In addition, the interferon elicited by OT in BCG-infected mice is shown to be a distinct molecular species of viral inhibitor and is designated “Type II interferon.” The designation “Type I interferon” is used to describe the viral inhibitor produced in BCG-infected or control mice given nonspecific stimuli such as bacterial lipopolysaccharide (LPS) or Newcastle disease virus (NDV). Although Type II interferon possesses the biologic attributes required to classify a viral inhibitor as “interferon,” it, as well as MIF, differs markedly from Type I interferon in stability at pH 2 and at 56°C, range of species specificity, and antigenicity. Antibodies against highly purified L cell interferon induced by NDV (anti-Type I interferon antibodies) neutralize the activity of interferons elicited in mice by nonspecific stimuli such as LPS or NDV. In contrast, the antiviral activity of Type II interferon and the macrophage-inhibitory activity of MIF, elicited by specific antigen in mice with delayed hypersensitivity, are not affected by high concentrations of antibody against Type I interferon. Evidence is presented which emphasizes the striking similarities in properties of MIF and Type II interferon, although it is not possible to determine whether the two mediators are the same or different protein molecules.
Mapping the H-2-Linked Genes Governing, Respectively, the Immune Responses to a Glutamic Acid-Alanine-Tyrosine Copolymer and to Limiting Doses of Ovalbumin
Authors: Dunham Edward K. , Dorf Martin E. , Shreffler Donald C. , Benacerraf Baruj
Published: December 2022

Addendum. Since these experiments were completed, mice of the strains A.TA and A.TB have become available, and their capacity to respond to GAT has been determined. Both these strains are recombinants within the H-2 complex between A.CA (H-2f, responders to GAT) and A.TH (H-2th, nonresponders to GAT) and carry the H-2 haplotype KfSs-SlpthDth. Groups of four of five mice of the recombinant and parental strains were immunized with GAT as before except that the second injection was given on day 21 and the secondary bleeding taken on day 28. Means and standard errors of the per cent of antigen bound by 1:5 dilutions of the secondary plasma samples were determined as before with the following results: A.CA, 55 ± 15 (three animals); A.TH, 9 ± 9 (four animals); A.TA, 87 ± 3 (three animals); and A.TB, 79 ± 3 (four animals).
The Cellular Basis for Tolerance or Immunity to Bovine γ-Globulin in Mice
Authors: Das Saroj , Leskowitz Sidney
Published: December 2022

BALB/c and DBA/2 mice were shown to have markedly different susceptibility to tolerance induction with bovine γ-globulin (BGG), the latter strain becoming tolerant at doses greater than 0.2 mg, while the former required more than 20 mg. When cells from BALB/c spleens were transferred to lethally irradiated DBA/2 recipients, the animals behaved like the normal DBA/2 in respect to tolerance induction, i.e., doses of 0.2 mg or more of BGG readily produced unresponsiveness. Similarly, when DBA/2 spleen cells were transferred to irradiated BALB/c recipients, behavior toward tolerance induction was like normal BALB/c, i.e., tolerance was not produced with doses less than 20 mg.

Analysis of response to SIII and typing of T cells by appropriate antisera showed that recipients were true chimeras in respect to B and T cells.

It was felt that the behavior of these irradiated reconstituted animals depended on the existence of a radiation-resistant macrophage which, in the case of BALB/c recipients, had the capacity to process small amounts of immunogenic material in BGG to make an immune response before concurrent induction of tolerance by monomeric BGG was complete. This was borne out by the greater capacity of BALB/c mice to filter out immunogenic material in BGG passaged in vivo.
Tolerance and Contact Sensitivity to Dnfb in Mice
Authors: Phanuphak Praphan , Moorhead John W. , Claman Henry N.
Published: December 2022

Tolerance and sensitivity to a potent contact sensitizer were studied in vivo and in vitro.

Contact hypersensitivity to 2,4-dinitro-1-fluorobenzene (DNFB) was induced in BALB/c mice by two daily paintings with 0.5% DNFB. The degree of hypersensitivity was assayed 4 days after the second painting by measuring the increments of ear thickness 24 hr after challenging the ears with 0.2% DNFB. Ear swelling was maximum at 24 hr and the histology was compatible with delayed type hypersensitivity.

Specific tolerance to DNFB was produced by a single i.v. injection of 15 mg of 2,4-dinitrobenzene-1-sulfonic acid sodium salt (DNB·SO3Na) 7 days prior to sensitization. DNB SO3Na given 2 days before, the same day as or 1 or 3 days after sensitization resulted in only partial unresponsiveness. Tolerance and sensitization were also measured by in vitro stimulation of DNA synthesis in cultured lymphoid cells. Lymph node cells taken 5 days after sensitization were specifically stimulated by DNP-BSA (but not by BSA) in vitro as assayed by increase of 3H-thymidine uptake. This response was dependent on the presence of T cells. Cells from non-sensitized mice responded poorly to DNP-BSA stimulation. Lymph node cells from tolerant mice responded 71 to 87% less than did sensitized cells. The results are discussed in terms of possible mechanisms of tolerance. In the context of this model of contact sensitivity, this appears to be tolerance at the T-cell level.
Suppressive Effects of in Vivo Immunization on PHA Responses in Vitro
Authors: Gershon Richard K. , Gery Igal , Waksman Byron H.
Published: December 2022

The PHA response of spleen cells from mice immunized with heterologous γ-globulin, serum albumin or erythrocytes 1 to 2 days previously may be severely impaired. The impairment disappears within a week. Inclusion of the specific antigen in culture can cause further suppression or unresponsiveness. The suppression of the PHA response was increased by increasing the immunizing dose of antigen, the amount of antigen in culture, the amount of PHA used for stimulation, the number of cells in culture, the use of an adjuvant in immunization, and the use of more stimulatory sera in culture. The PHA response of spleen cells from mice given a tolerogenic form of bovine γ-globulin (BGG) was suppressed in the presence of BGG, although less so than that of mice given an immunogenic form of the antigen. The tolerogenic form was, however, more suppressive when added to the culture. The ability of a relatively small number of antigen reactive cells to markedly affect the PHA response suggests that suppressive cell interactions play an important causative role.
Lymphocyte Plasma Membranes
Authors: Zimmerman Barry
Published: December 2022

Plasma membranes prepared from rat thymocytes were used to immunize rabbits and after 6 months the resulting sera had cytotoxic titers of 1:1280 to 1:2560 for rat thymocytes. With the lactoperoxidase-catalyzed iodination reaction the thymus plasma membranes were labeled with 125Iodine, solubilized in non-ionic detergent, and immuno-precipitated with the cytotoxic rabbit anti-rat thymus sera. These sera bound 3.5 to 4.4% of the total acid insoluble counts while control sera bound 0.6%. Subsequently, the precipitates were solubilized, reduced, and characterized on SDS containing polyacrylamide gels. The labeled antigenic component proved to have a m.w. ∼45,000. The same antigenic material could be labeled by in vitro incubation of rat thymus cells in 14C amino acids. Absorption studies suggested that this antigenic membrane constituent was shared by all lymphoid cells and represented a major immunodominant area on lymphocyte surfaces during heterologous immunization.
Immunopathologic Aspects of Infection with Lagos Bat Virus of the Rabies Serogroup
Authors: Tignor Gregory H. , Shope Robert E. , Gershon Richard K. , Waksman Byron H.
Published: December 2022

After intracerebral inoculation of Lagos bat virus, immunosuppression by either Cytoxan or thymectomy and sublethal irradiation prolonged survival and decreased the incidence of paralytic signs in random-bred Charles River mice. A similar effect after Cytoxan treatment was observed in BALB/c mice, but not in C57BL/6, or A strain mice. Weanling hamsters treated with Cytoxan showed only a decrease in the incidence of paralytic signs. The administration of Lagos bat virus antibody to immunosuppressed Charles River mice on day 8 after inoculation of a high dose of virus was followed by rapid paralysis and early death. Preliminary attempts to apply this model to rabies virus indicate that biologic differences among strains of rabies may be an important determinant for detection of the measurable effect of immunosuppression.
The Effect of Metabolic Inhibitors and Hydroxylamine on the Immune Response in Mice to Mycobacterial Ribonucleic Acid Vaccines
Authors: Youmans Anne S. , Youmans Guy P.
Published: December 2022

Metabolic inhibitors were used in an attempt to determine the manner by which mycobacterial ribonucleic acid (myc. RNA) immunizes against tuberculosis. Inhibitors of protein synthesis (streptomycin sulfate, chloramphenicol, cycloheximide) and inhibitors of DNA-dependent RNA polymerase (rifampin) had no effect on the immune response of mice vaccinated with myc. RNA. All of these compounds except streptomycin sulfate, however, inhibited antibody formation under the same conditions. In contrast, inhibitors of deoxyribonucleic acid (DNA) synthesis significantly reduced the immune response. Five-fluoro-deoxyuridine (FUDR), which inhibits thymidine synthetase, when injected just before vaccination, reduced the immune response but had no effect if injected 18 hr after vaccination. These results indicated that early synthesis of DNA was important, and that DNA once formed was not affected by FUDR. Ethidium bromide, proflavine and chloroquine were each mixed with myc. RNA and injected into mice. Under these circumstances the immune response was significantly lowered. In addition, chloroquine was injected at various times and significant reduction in the immune response occurred only if it were injected 1 hr or later after vaccination. This compound binds with DNA, and shows again the necessity for the presence of DNA in the immune process. Actinomycin D injected at the time of vaccination or 1 hr after vaccination had no effect on the immune response. However, if actinomycin D was injected 18 hr after vaccination or injected daily for 4 days after vaccination, a marked reduction in the immune response occurred. These results indicated that actinomycin D, which inhibits RNA transcription may have inhibited the replication of myc. RNA. A mutogenic agent, hydroxylamine, also reduced significantly the immunogenic activity of myc. RNA. These results closely paralleled those obtained by other investigators in studies aimed at eliciting the mode of replication of oncogenic viruses. The replication of these RNA tumor viruses is unique in that DNA synthesis is required for viral RNA replication early after infection, and there is a requirement for RNA transcription. The similarity between the manner by which myc. RNA immunizes and RNA tumor viruses replicate is discussed, and a comparison also is made of the two diseases, tuberculosis and tumors formed by oncogenic viruses. These results strongly suggest that myc. RNA may immunize against tuberculosis by forming a DNA template in the host cell which then transcribes specific information for the replication of myc. RNA, and thereby transforms a normal cell into a specific immune cell. If correct, this would provide a new concept of the nature of cellular immunity of tuberculous infection.
Immunofluorescence Studies on Thymocytotoxic Antibody from New Zealand Black Mice
Authors: Parker Leroy M. , Chused Thomas M. , Steinberg Alfred D.
Published: December 2022

Natural thymocytotoxic antibody (NTA) which occurs in high titer in New Zealand Black (NZB) mice was compared with AKR anti-θ (C3H) antiserum for ability to induce redistribution of C57BL/6 thymocyte surface antigen. NTA reacted with both thymocytes and brain cells bearing either the θ-AKR or θ-C3H alloantigens and was found to be an IgM antibody. Anti-θ (C3H) was purified to obtain the IgG fraction which was used in these studies. Neither NTA nor anti-θ antibody induced cap formation alone, however, both formed caps upon the addition of specific anti-immunoglobulin sera. Studies involving the sequential exposure of thymocytes to both antibodies demonstrated that θ and NTA reactive antigen co-migrate on the thymocyte surface. These studies suggest that NTA reacts with a determinant common to both the θ-AKR and θ-C3H alloantigens.
Comparative Studies of Immunoglobulin Receptors and Antibody-Dependent Cell Cytotoxicity (ADCC) in Rat Lymphoid Organs
Authors: Kedar Eli , de Landazuri Manuel Ortiz , Fahey John L.
Published: December 2022

The capability of rat lymphoid cells to destroy antibody-coated target cells (antibody-dependent cell cytotoxicity, ADCC) and to form rosettes with immunoglobulin-coated erythrocytes (EA) was compared. Because both processes require binding of a lymphoid cell to antibody associated with cellular antigen, we undertook to determine if both assays detect the same cell population. Peripheral blood lymphocytes, spleen and lymph node cells had both activities, but there was no correlation between the number of rosettes and the ADCC activity. Blood lymphocytes had the highest cytotoxic potential, while spleen cells produced the highest percentage of rosettes. Bone marrow cells were devoid of cytotoxic activity though many of the cells were able to form rosettes. Thymus cells were neither cytotoxic nor able to form rosettes. Following treatment of spleen cells with 1) anti-rat Ig serum, 2) high concentrations (τ1 mg/ml) of trypsin and Pronase (but not papain), or phospholipase-C, the number of rosettes was strongly diminished, whereas the cytotoxic activity was unimpaired. The ADCC activity was not altered after removal of 60% of the rosetteforming cells by nylon column, but was greatly reduced when 85% of the rosette-forming cells were removed by anti-SRBC serum.

These observations indicate that: 1) there is no correlation between the percentage of EA rosette-forming cells and the ADCC activity in various lymphoid tissues, 2) these activities were not equally susceptible to enzymes' action, 3) the cytotoxic activity is mediated by a subpopulation of the EA rosette-forming cells, and 4) probably fewer immunoglobulin receptor sites per unit surface membrane are required for ADCC cytotoxicity than for rosette formation.
Induction of Tolerance to Coccidioidin in Newborn Guinea Pigs
Authors: Ibrahim Ali Bin
Published: December 2022

The tolerogenic effect of the coccidioidal antigens was studied by injecting newborn guinea pigs, within 24 hr of birth, with killed hyphal cells of Coccidioides immitis in complete Freund's adjuvant (CFA). Such animals, when injected as adults with killed hyphal cells of C. immitis in CFA, did not respond with delayed hypersensitivity (DH) to coccidioidin as did other normal animals of the same age group, but they did exhibit DH to tuberculin. The tolerance to coccidioidin lasted for 3 to 6 months. Injection of newborns as late as 3 days after birth also induced tolerance which lasted for 30 to 60 days. Peritoneal macrophages from the coccidioidin-tolerant animals migrated normally in the presence of coccidioidin but were inhibited by tuberculin or PPD. The adoptive transfer of coccidioidin sensitivity to tolerant animals by intraperitoneal injection of peritoneal exudate cells from sensitized adult guinea pigs was possible. This transferred sensitivity was temporary and lasted for 3 to 5 days.
Glomerulonephritis in the Graft-vs-Host Reaction: Serologic Demonstration of Anti-Host Antibodies
Authors: Kano Kyoichi , Beldotti Lorraine , Milgrom Felix , Schwartz Robert S.
Published: December 2022

Antibodies against C57BL/6 cells were detected in serum and renal eluates of (C57BL/6 × DBA/2)F1 mice that developed nephritis after receiving injections of DBA/2 spleen cells. This model of glomerulonephritis provides an example of immune complex deposits consisting of histocompatibility antigens and antibodies against histocompatibility antigens.
Purification of Human T and B Lymphocytes
Authors: Greaves Melvyn F. , Brown Geoffrey
Published: December 2022

Analysis of immune responses involving distinctive cell types has been greatly facilitated by the development of technology for separating and purifying different lymphocyte populations (1–5). In conjugation with the use of markers which identify selective expression of cell surface components (6, 7) this approach provides an efficient means of studying multi-component immune responses. Marker systems identifying human thymus derived2 (T) and bone marrow-derived (B) lymphocytes are now well advanced (reviewed in 8–10) and we report here the application of various markers to determine the efficacy of separation techniques aimed at providing a simple and reproducible protocol for the preparation of effectively pure human T and B lymphocytes for functional studies.

Materials and Methods. All cell separations were performed on human tonsil cell suspensions since these were available daily from University College Hospital ENT Theatre and provided a convenient source of over 2 × 108 viable lymphocytes per tonsil.
Preparation and Properties of Anti-Sulfatide Serum
Authors: Hakomori Sen-Itiroh
Published: December 2022

Antisera against sulfatide were prepared and their properties were examined. The specificity was directed toward the galactopyranosyl-3-sulfate residue.
Presence of Antibody-Forming Cell Precursors in the Mouse Adherent Spleen Cell Population
Authors: Loughman Barbara E. , Farrar John J. , Nordin Albert A.
Published: December 2022

It is generally accepted that primary in vitro immunization of mouse spleen cells involves the synergistic participation of at least three different cell types (for reviews see 1 and 2). The initial separation of mouse spleen cells into two populations on the basis of their selective adherence to plastic culture dishes was reported by Mosier (3). Neither the adherent nor the non-adherent sub-populations, when cultured in vitro, respond to sheep erythrocytes (SRC).1 When these two subpopulations are recombined, the response to SRC is restored. It was later established that the non-adherent cell population contains both bone marrow- (B) and thymus- (T) derived lymphocytes. Although the non-adherent population contains T-helper cells and precursors to the plaque-forming cells (P-PFC), its nonresponsiveness is due to the lack of accessory cells. Restoration of the responsiveness of non-adherent cells by the addition of adherent cells has led to the assumption that the adherent cell monolayer provides little other than accessory cell function.
Protection against Autoimmune Thyroiditis in Guinea Pigs by Thyroglobulin Antiserum
Authors: Sharp Gordon C. , Wortis Henry H. , LaRoque Robert L. , Mullen Helen , Kyriakos Michael
Published: December 2022

Repeated administration of hyperimmune rabbit antiserum to guinea pig thyroglobulin (GPTG) consistently protected guinea pigs from developing autoimmune thyroiditis when studied 23 days following active immunization with GPTG in complete Freund's adjuvant. Rabbit antisera to various non-thyroid antigens were much less effective in causing inhibition. The suppressive effect noted with antiserum to keyhole limpet hemocyanin (KLH) differed from anti-GPTG suppression in that it was transient and associated with reduced delayed hypersensitivity to GPTG. Protective effects of anti-GPTG persisted at 156 days even in the presence of intact delayed hypersensitivity to GPTG. Transfer of hyperimmune anti-GPTG during the first 10 days after active immunization still resulted in protection, but hyperimmune anti-GPTG given late or early anti-GPTG given throughout the experiment failed to protect.
Antisera to Human Cytomegaloviruses Prepared in the Guinea Pig: Specific Immunofluorescence and Complement Fixation Tests
Authors: Huang Yung-Tsun , Huang Eng-Shang , Pagano Joseph S.
Published: December 2022

Specific, high titered antisera to human (C-87 and AD-169) and simian (GR-2757) cytomegaloviruses were prepared in guinea pigs and used in complement fixation (CF) and immunofluorescence (IF) tests. There were no detectable cellular antibodies in the antisera. Both the CF and the IF tests revealed that the two human strains are closely related or identical and have little if any interspecies cross-reactivity with the simian strain.
Immuno-Regulatory Role of Spleen Localizing Thymocytes
Authors: Gershon Richard K. , Lance Eugene M. , Kondo Kazunari
Published: December 2022

After inoculation into lethally irradiated F1 mice, parental thymocytes reacting to host antigens, synthesize deoxyribonucleic acid in the spleens and lymph nodes of the recipient mice. Removal of the spleen localizing fraction of the thymocyte inoculum, by splenectomizing recipients 3 hours after cell injection, markedly alters the response of the cells in the lymph nodes without affecting the number of chromium 51 labeled cells which localize in the nodes. The effects of splenectomy may be bidirectional; increases or decreases of the lymph node response may ensue. At higher thymocyte doses splenectomy leads to an increased lymph node response, apparently by diminishing suppressive interactions among the inoculated cells. When spleen removal increases the response of the cells in the lymph nodes, reinjection of the cells removed with the spleen suppresses the response. The addition of F1 thymocytes to the inocula of parental cells can alter the response of the parental cells. Such effects of the F1 cells, which occur in the recipients lymph nodes, can be prevented by removal of the spleen localizing fraction of the thymocyte inoculum. Delaying splenectomy by 24 to 48 hr diminishes its effect. We suggest spleen localizing T cells may play an important role in immuno-regulation.
Identification of Cells Infiltrating the Minor Salivary Glands in Patients with Sjögren's Syndrome
Authors: Chused Thomas M. , Hardin John A. , Frank Michael M. , Green Ira
Published: December 2022

Patients with Sjögren's syndrome (SS) have mononuclear cell infiltration, tissue destruction and eventual fibrosis of the salivary and lacrimal glands. The type of mononuclear cell infiltrating the minor salivary glands of the lip in three patients with SS has been determined. Sheep erythrocytes coated with 19S antibody and activated third component of complement were found to adhere to human bone marrow-derived (B) cells and monocytes in tissue sections. Erythrocytes coated with 7S antibody adhered only to monocytes. The localization of these reagents in tissue sections was used to demonstrate that the major portion of mononuclear cells infiltrating SS salivary tissue are B cells and that they are arranged in a particular way in the infiltrate.
Polyribitol-Phosphate: An Antigen of Four Gram-Positive Bacteria Cross-Reactive with the Capsular Polysaccharide of Haemophilus Influenzae Type B
Authors: Argaman Meir , Liu Teh-Yung , Robbins John B.
Published: December 2022

Polysaccharide extracts of whole bacteria of the species Staphylococcus aureus, Bacillus pumilus, Lactobacillus plantarum and Streptococcus faecium were found to precipitate with Haemophilus influenzae type b (HITB) antiserum. This precipitation was due to the polyribitol-phosphate component of these bacteria reacting with the type b capsular polysaccharide antibodies of the HITB antiserum. Immunization of rabbits and burros with formaldehyde-fixed cross-reacting bacteria and blood-borne infections in humans caused by S. aureus (coagulase positive) induced precipitating and bactericidal HITB antibodies. Preliminary analysis of the HITB capsular polysaccharide revealed ribitol in addition to ribose providing an explanation for this cross-reaction. The ubiquitous distribution of Gram-positive bacteria containing polyribitol-phosphate in their cell-wall teichoic acid suggests that these bacteria may be responsible in part for the wide prevalence of antibodies to HITB capsular polysaccharide in the normal human population.
Specificity of IgM and IgM Antibodies after Challenge with Antigenically Related Togaviruses
Authors: Westaway E. G. , Della-Porta A. J. , Reedman Beverley M.
Published: December 2022

Rabbits were immunized with a group A or a group B togavirus and challenged 6 weeks later with a related togavirus from a different subgroup. IgG antibodies in antisera cross-reacted broadly at all times in hemagglutination-inhibition tests with group B viruses whereas within group A they were specific for the primary virus until challenge after which they reacted with both primary and challenge viruses. In contrast, IgM antibodies were clearly specific within both groups for the most recent immunizing virus; however, IgM memory was expressed when the challenge and primary viruses belonged to the same antigenic subgroup. The antigenic composition of both groups of togaviruses is discussed in relation to their envelope proteins, and hypotheses are proposed to account for the antibody specificities of the observed immunoglobulin responses.
Similarity of Migration Inhibitory Factor(s) Produced by Human Lymphoid Cell Line and Phytohemagglutinin and Tuberculin-Stimulated Human Peripheral Lymphocytes
Authors: Papageorgiou Photini S. , Sorokin Carol F. , Glade Philip R.
Published: December 2022

The migration inhibitory factor(s) (MIF) derived from the supernatants of an established human lymphoid cell line (PGLC-33H) and from human peripheral lymphocytes stimulated in vitro by phytohemagglutinin (PHA) and tuberculin (PPD) were characterized and compared. Separation of these supernatants by differential ultrafiltration showed peak inhibitory activity for all three in the fraction containing molecules of the size 10,000 to 50,000. Separation by Sephadex gel filtration (G-75) revealed similar elution profiles for these MIF. Peak inhibitory activity eluted between enzyme markers of m.w. 17,000 and 25,000. The MIF activity contained in these supernatants was found to be heat stable (56°C for 30 min) and chymotrypsin sensitive. The MIF derived from human long term lymphoid cell lines appears to be similar to the MIF produced by human peripheral lymphocytes stimulated in vitro by mitogen and antigen.
Lymphocyte Interferon Production and Transformation after Herpes Simplex Infections in Humans
Authors: Rasmussen Lucille E. , Jordan George W. , Stevens David A. , Merigan Thomas C.
Published: December 2022

Antigen-stimulated lymphocyte transformation, as measured by 3H-thymidine incorporation, and interferon production were studied serially in humans after Herpes virus hominis (HSV) disease. Lymphocytes were isolated by filtration through a nylon fiber column and added to isologous macrophage cultures prepared 1 week previously. Macrophage-lymphocyte interaction was required for optimum interferon production. A selective increase in interferon production was associated with disease occurrence in some individuals. The increase in interferon production by lymphocytes was maximum between 2 and 6 weeks after disease. In contrast, lymphocyte transformation and serum antibody levels were similar throughout a 12-week period after disease and appeared to reflect prior immune status rather than acute infection. Some subjects with HSV disease failed to produce a detectable interferon response within the 2- to 6-week period after disease. In these individuals, lesion recurrence was more frequent when compared to subjects producing greater quantities of interferon after disease.
Regulation of Homocytotropic Antibody Formation in the Rat
Authors: Okumura Ko , Tada Tomio
Published: December 2022

Chemical and physicochemical properties of the previously reported “antigen-specific T cell factor” that negatively regulates hapten-specific homocytotropic antibody (HTA) response in the rat were investigated. This factor, which is extractable from mechanically disrupted thymocytes obtained from rats hyperimmunized with dinitrophenylated Ascaris suum extract (DNP-As), showed a strong inhibitory effect on hapten-specific HTA formation in recipient animals that had been x-irradiated and immunized with DNP-As so as to produce high and persistent HTA formation. The treatment of the thymocyte extract with proteinases and nucleases revealed that the active factor is not digestible by DNase and RNase, but is destroyable by trypsin and Pronase, indicating the protein nature of the T cell factor. In sucrose density gradient ultracentrifugation, the activity sedimented slower than IgG hemagglutinin. In gel filtration with Sephadex G-100, the inhibitory activity was not detected in the fraction eluted with a void volume, but was found in a fraction eluted in the range between about 35,000 and 60,000 daltons. By electrophoresis in agar the activity was located in regions corresponding to rat α- and β-globulins but not in regions corresponding to albumin and γ-globulin. These results indicate that the antigen-specific inhibitory T cell factor is a protein with a smaller molecular weight than other immunoglobulins, further supporting previous findings that the T cell factor is not a usual immunoglobulin but may be a unique receptor of T cells.
Antibody-Mediated Target Cell Lysis by Nonimmune Cells: The Use of Anti-Immunoglobulin to Distinguish Effector Cell Populations
Authors: Resch Klaus , Gelfand Erwin W. , Prester Marlot
Published: December 2022

The role of a goat IgG anti-rabbit immunoglobulin on antibody-mediated target cell lysis by nonimmune cells was investigated. Using a homologous system of rabbit effector cells and rabbit anti-fowl erythrocyte serum, two populations of cells effective in the lysis of antibody-coated target cells could be differentiated by anti-immunoglobulin. In the presence of the anti-immunoglobulin, the cytotoxicity caused by phagocytic cells was suppressed. The IgG or (Fab′)2 fraction of the anti-immunoglobulin enhanced target cell destruction initiated by purified lymph node and spleen lymphocytes and to a lesser extent that initiated by purified thymus cells. Monovalent (Fab) anti-immunoglobulin decreased the cytotoxic activity of all effector cells. The mechanism of enhancement appeared to be due to cross-linking between effector lymphocytes and antibody-coated target cells by the divalent anti-immunoglobulin.
Interaction of Anti-Thymocyte Serum with Hematopoietic Stem Cells
Authors: Mookerjee B. K. , Azzolina Luigi , Poultar Lyndall
Published: December 2022

In vitro incubation of spleen cell suspensions in dilutions of anti-thymocyte globulin (ATG) leads to abrogation of the capacity of such cells to mount a graft-vs-host reaction; however, such incubation leads to considerable inactivation in the content of the hematopoietic stem cells. By prior absorption of the ATG preparation with mouse liver or spleen cells, it was found possible to eliminate the toxicity of ATG toward hematopoietic stem cells without any reduction in its immunosuppressive potency. Administration of ATG to intact animals was found to stimulate endogenous colony formation. Intravenous injection of ATG led to early and significant elevation in the serum level of colony-stimulating factor in the injected animals when compared to that after similar injection of normal horse globulin. Short or long term in vivo administration of anti-thymocyte serum did not lead to reduction in the number of spleen-colonizing stem cells in the bone marrow of the treated animals or in the capacity of such cells to reconstitute lethally irradiated syngeneic animals.
Interaction of Complement with Antigen-Antibody Complexes of Rabbit Allotypic Antigens and Their Corresponding Antibodies
Authors: Kakinuma Mitsuaki
Published: December 2022

Multiple precipitin lines are observed in agar gel when rabbit immunoglobulin possessing α-locus determinants is allowed to react with its corresponding antiserum in double diffusion tests. One of these lines was proven to be complement-dependent. Coexistence of complement-dependent and complement independent precipitin lines in this particular allogenic system suggests that heterogeneity existed with respect to the avidity of antibodies direct to defined determinant groups.
Effect of Rifamycin-SV Derivatives on Immunologic Responses in Culture
Authors: Osoba David
Published: December 2022

Both rifampicin and 4-N-demethyl rifampicin (AF/AP) inhibited the formation of plaque-forming cells to sheep erythrocytes at concentrations that did not affect the formation of myeloid colonies in culture. Two other rifamycin-SV derivatives, 4-N-benzyl demethyl rifampicin (AF/ABP) and 2,6-dimethyl-4-benzyl-4-demethyl rifampicin (AF/ABDP CIS) had no significant effect on either the formation of plaque-forming cells or myeloid colonies. Rifampicin and AF/AP also inhibited the incorporation of tritiated thymidine in one-way mixed leukocyte reactions both by mouse spleen cells and by human peripheral blood leukocytes, and AF/AP markedly inhibited the generation of cytotoxic effector cells in the mixed leukocyte reaction by mouse spleen cells. The action of AF/AP on the formation of plaque-forming cells appears to affect an earlier cellular function than does rifampicin. Although the precise nature of the pharmacologic effect of these drugs on the formation of antibody-forming cells and on the generation of cytotoxic effector cells is unknown, the inhibitory effect of at least some of the members of this family of drugs on viral RNA-dependent DNA polymerase (reverse transcriptase) suggests that reverse transcriptases may be involved in antibody formation by normal cells of the immunologic system.
Comparison and Yield of Antigen- or Mitogen-Induced Human Lymphotoxins
Authors: Boulos Galal N. , Rosenau Werner , Goldberg Melvin L.
Published: December 2022

The yield of lymphotoxin (LT) in cultures of human lymphocytes stimulated with a mitogen, phytohemagglutinin (PHA), is about 60 times greater than that obtained by antigenic stimulation with mixed lymphocyte cultures. Lymphotoxin induced under both conditions behaved identically when we employed electrophoresis on 9% acrylamide gel (pH 8.6 or 7.4), ultracentrifugation in 3 to 10% sucrose density gradients, and testing for pH stability. Both LT were neutralized in like degree by the addition of anti-LT serum against purified PHA-induced human LT. Thus, utilizing multiple analytical methods based on diverse principles, we found no differences in the two LT, strongly implying that both types are indeed identical. The two major implications of this conclusion are: 1) Although under natural in vivo conditions, only antigen-induced LT would be operative, in the light of our findings, mitogen-induced LT may also be used in biologically meaningful experiments. 2) Furthermore, the larger yield of mitogen-induced LT represents a great advantage in purification of the lymphocyte mediator present in the starting material only in small amounts. In addition, it is of interest that 89% of the cytotoxic activity of crude LT induced by PHA stimulation can be neutralized by antiserum prepared against purified LT, an indication that the bulk of the cytotoxicity in crude LT appears to be due to the cytotoxin comprising our purified LT preparation.