bluuebunny/crossref_metadata_2025 huggingface dataset:
Complement Inhibitor(s) Released by Leukocytes
Authors: Bernard Alain , Boumsell Laurence , Borsos Tibor , Good Robert A. , Day Noorbibi K.
Published: December 2022

Sheep erythrocytes pretreated with supernatants of mouse spleen or thymus cells become resistant to lysis by guinea pig complement. The inhibitory activity (IA) reduces the utilization of C2 by EAC. Because IA binds to the surface of sheep erythrocytes and does not inhibit C1̄ irreversibly, it is probably a hitherto undescribed inhibitor of complement.
Complement Inhibitor(s) Released by Leukocytes
Authors: Bernard Alain , Boumsell Laurence , Borsos Tibor
Published: December 2022

Mouse spleen or thymus cells in short-term culture release a factor, designated S, that binds to sheep erythrocytes (E). Supernatant-treated sheep erythrocytes (SE) are capable of fixing and transferring the activated first component of guinea pig complement. SEC1̄, however is not capable of initiating hemolysis by the rest of the complement components. SE is capable of binding but not activating native C1; native C1 bound to SE seems irreversibly inhibited. Evidence is presented that S may be the same factor as the previously described inhibitor released by mouse spleen or thymus cells that inhibits the utilization of C2 by EAC1̄4.
Carrier-Induced Tolerance to Nucleic Acid Antigens
Authors: Stollar B. David , Borel Yves
Published: December 2022

BALB/c and SJL mice were treated with nucleosides-IgG1 as a tolerogen, before either primary or secondary immunization with nucleosides-keyhole limpet hemocyanin. Nucleoside-specific responses were measured serologically by a modified Farr assay, with either 14C-labeled denatured DNA or nucleosides-131-I-labeled BSA as test antigen. Specificity of the response was tested by hapten inhibition experiments.

Multiple doses of nucleosides-IgG1 tolerogen given before the primary or secondary immunization effectively suppressed the secondary and tertiary anti-nucleoside responses. The tolerogen did not suppress the response to an unrelated hapten-KLH conjugate. The IgG alone did not suppress the anti-nucleoside response of BALB/c mice to nucleosides-KLH. Single doses of tolerogen before the primary or secondary immunization were less effective. Residual antibody in partially suppressed BALB/c mice showed changes in specificity as compared to controls. Suppression of the secondary response of SJL mice was measured much more readily by binding of nucleosides131-I-BSA than by binding of denatured DNA. This reflected an altered specificity of the residual antibody; in control animals, antibodies were directed against all four nucleosides, whereas the antibodies of partially suppressed animals were directed only against guanosine.

Suppression of anti-nucleic acid antibody responses may have therapeutic application in the management of systemic lupus erythematosus.
Temporal Dependence of the Inhibition by Cytochalasin B of Antigen-Initiated Migration Inhibition Factor Production
Authors: Sasson S. Z. Ben , Rosenthal Alan S.
Published: December 2022

The effects of cytochalasin B on the antigen recognition process have been examined using an in vitro assay of macrophage-inhibition factor (MIF) production by activated guinea pig T lymphocyte-enriched populations. The data indicate that cytochalasin B inhibits MIF production if added at the initiation of culture but not if added 2 hr after antigen. These data are interpreted as consistent with either an inhibition by cytochalasin B of the physical association of antigen-bearing macrophage and lymphocytes or with an inhibition of the access of antigen to the T lymphocyte receptor.
Immunity to Toxoplasma Gondii Induced in Vitro in Non-Immune Mouse Macrophages with Specifically Immune Lymphocytes
Authors: Sethi Krishan K. , Pelster Bernd , Suzuki Naoyoshi , Piekarski Gerhard , Brandis Henning
Published: December 2022

Male and female CBA mice were used to study in vitro the mechanisms involved in the development and expression of cellular immunity to toxoplasma infection. The lag phase preceding toxoplasma division was delayed in nonimmune macrophages obtained from peritoneal cavities stimulated with thioglycollate. Specific anti-toxoplasma activity was conferred on nonimmune macrophages incubated with toxoplasma-immune spleen lymphocytes and soluble toxoplasma antigen. Treatment of immune spleen cell populations with anti-θ serum plus complement abolished completely their activity of conferring anti-toxoplasma activity on nonimmune macrophages, demonstrating that the essential cells were T lymphocytes. The mediator(s) responsible for the acquisition of immunity to toxoplasma in the nonimmune macrophages were soluble. Heat-inactivated, toxoplasma-immune serum rendered toxoplasmas incapable of infecting nonimmune macrophages of fibroblasts. The findings are related to previous investigations of induced immunity in animals and man.
IgG-Dependent Clearance of Red Blood Cells in IgG Myeloma Patients
Authors: Scornik Juan Carlos , Salinas Mario Cesar , Drewinko Benjamin
Published: December 2022

Red blood cells sensitized with IgG antibodies are rapidly cleared from the circulation of myeloma patients, in spite of serum IgG concentrations several times higher than normal. In contrast, in vitro phagocytosis and cytotoxicity of IgG-sensitized erythrocytes are inhibited by dilutions of normal fresh serum. It is concluded that the current explanations derived from in vitro experiments are insufficient to explain the IgG-dependent clearance of red blood cells.
Specific Partial Depletion of Graft-vs-Host Activity by Incubation and Centrifugation of Mouse Spleen Cells on Allogeneic Spleen Cell Monolayers
Authors: Mage Michael G. , McHugh Louise L.
Published: December 2022

Spleen cells (from BALB/c mice immunized with the C57BL/6 lymphoma EL4, or from non-immune BALB/c) were incubated on monolayers of [C57BL/6 × BALB/cF1 (B6CF1) spleen cells on polylysine-coated polystyrene Petri plate, for ½ hr or for 1 hr at 37°C followed by centrifugation of the monolayers for 5 min at 70 × G to 110 × G at 34 to 37°C. Control monolayers were BALB/c spleen cells. As measured by the Simonsen spleen weight assay in neonatal mice, graft-vs-host (GVH) activity was partially depleted in cell populations nonadherent to B6CF1 monolayers. Residual GVH activity of these nonadherent cells was about half that of cells incubated on the control syngeneic monolayers (the mean of eight experiments was 49% ± 11% S.D.). Two or three consecutive cycles of incubation and centrifugation did not significantly diminish the residual GVH activity, suggesting that spleen cells with GVH activity are heterogeneous with respect to binding to allogeneic target cells under the above conditions. Cell populations nonadherent to third-part [A × AL]F1 monolayers retained full activity, and cell populations partially depleted of GVH activity in B6CF1 neonates had full activity in third-party [BALB/c × AL]F1 neonates.
Structural and Genetic Basis of the in Vivo Immune Response to TNP-LPS
Authors: Jacobs Diane M.
Published: December 2022

TNP-lipopolysaccharide (TNP-LPS) is a potent T-independent antigen in vivo, inducing a TNP-PFC response in T-depleted animals. The structural integrity of the lipid A-KDO portion of the LPS carrier molecule appears to be required since the haptenated LPS from Salmonella minnesota Re595 is immunogenic whereas the haptenated derivative of base hydrolyzed LPS is not. The immune response is not associated with any of the common histocompatibility types, but does depend on the ability of the host strain to respond to LPS. C3H/HeJ mice are not killed by low doses of LPS and give a poor PFC response to TNP-LPS. Lethality and immunogenicity are dominant responses in hybrids of C3H/HeJ and responder mice. The structural and genetic requirements for the response to TNP-LPS suggest an active role for the carrier in the immunogenicity of this T-independent antigen.
Identification and Properties of J Chain Isolated from Catfish Macroglobulin
Authors: Mestecky Jiri , Kulhavy Rose , Schrohenloher Ralph E. , Tomana Milan , Wright Genesis P.
Published: December 2022

After the cleavage of disulfide bonds of macroglobulin isolated from channel catfish (Ictalurus punctatus), an electrophoretically fast-moving polypeptide, which resembled human J chain, was released. On a Sephadex G-200 column equilibrated in 5 M guanidine, the elution position of the J chain overlapped with the descending part of the L chain peak. Further purification was achieved by DEAE ion-exchange chromatography. The isolated polypeptide, which had a molecular weight of 14,800 ± 500, as determined ultracentrifugally by sedimentation equilibrium in 5 M guanidine, contained 7% carbohydrate with one residue of fucose, two of mannose, one of galactose, two of glucosamine, and one of sialic acid per chain. A comparison of catfish and human J chain amino acid analyses showed the former to have a higher content of serine, glycine, and phenylalanine and a lower content of aspartic acid, isoleucine, and arginine. Tryptic peptide maps of catfish and human J chains revealed very few common peptides. Rabbit and guinea pig antisera to human J chain did not cross-react with catfish J chain. Untreated, resuced and alkylated, S-sulfonated, or cyanogen bromide cleaved macroglobulin from the gar (Lepisosteus osseus) contained no polypeptide analogous to either catfish or human J chain by the criteria employed in this study.
Triggering Mechanism of B Lymphocytes
Authors: Watanabe Takeshi , Kishimoto Tadamitsu , Miyake Tadao , Nishizawa Yoshio , Inoue Hideo , Takeda Yoshiro , Yamamura Yuichi
Published: December 2022

In vitro stimulation of rabbit lymphocytes with anti-immunoglobulin antibody (anti-Ig) induced ornithine decarboxylase (ODC) (EC4.1.1.17) in the cells. The enzyme activity reached the maximum at 8 hr after stimulation of the cell with antibody and returned to the unstimulated level after 20 hr. The optimum concentration of antibody to produce the maximum activity of ODC was 25 µg/ml, which was in the same range that required for the maximum production of IgG and the highest rate of DNA synthesis. ODC was again induced with soluble factor (SF) in the cells that had been previously treated with anti-Ig antibody for 24 hr. The treatment of the cells with anti-thymocyte serum and complement did not affect the activity of the enzyme induced by anti-Ig or by SF, indicating that enzyme induction was an event occurring in B cells and not in T cells. The addition of mitotic inhibitors, such as hydroxyurea (HU) or cytosine arabinoside (Ara-C), to the culture with anti-Ig did not inhibit the production of ODC. The presence of cytochalasin-B with anti-Ig did not inhibit, but rather enhanced, the synthesis of ODC. On the other hand, the presence of HU or cytochalasin-B with soluble factor completely suppressed the induction of ODC. These results suggested that intracellular events that trigger the induction of ODC by the interaction of Ig receptor with anti-Ig may be different from the events that induce the same enzyme triggered by SF.
Receptors for IgG and Complement in Human Spleen Lymphoid Cells. Preferential Binding of Particulate Immune Complexes through Complement Receptors
Authors: Scornik Juan Carlos , Drewinko Benjamin
Published: December 2022

Human lymphoid spleen cells attached to Petri dishes by poly-l-lysine bind 51Cr-labeled erythrocytes coated with IgG antibodies or complement but not uncoated erythrocytes or those coated with IgM antibodies. The number of erythrocytes bound through complement receptors is several times larger than that bound through IgG receptors. Increasing up to five times the number of IgG molecules on the red blood cells only leads to a slight increase of binding. However, the addition of complement to the IgG-coated erythrocytes increases 10 times the binding to spleen cells, even in the presence of an excess of normal IgG. These results can be explained by postulating that there is a larger number (or greater affinity) of spleen cell receptors for complement than that of spleen cell receptors for IgG.
Lymphoid Cell Subpopulations
Authors: Howe Michael L. , Cohen Lewis
Published: December 2022

Mixtures of isogenic thymocytes (TC) and lymph node cells (LNC) were shown to exhibit synergistic responsiveness to M and H-2 alloantigens in the mixed lymphocyte interaction (MLI). With respect to the kinetics and magnitude of proliferation and effector cell generation, the response occurring in synergizing cultures closely resembled that of optimal numbers of LNC or spleen cells (SC). In addition, the antigen specificity of effector cells generated by synergizing cultures was similar to that of effectors derived from cultures containing optimal numbers of responding SC. LNC-TC mixtures also exhibited synergy in response to the phytomitogens concanavalin A and pokeweed mitogen but not to phytohemagglutinin. Weakly positive synergy was observed in response to bacterial lipopolysaccharide. It is proposed that the phenomenon of synergy is not restricted to cultures containing mixtures of LNC and TC but also occurs in cultures containing optimal numbers of LNC or SC as a result of interactions between subpopulations of lymphocytes contained within these tissues.
Immune Response to a Syngeneic Mammary Adenocarcinoma
Authors: Fortner G. William , Kuperman Oded , Lucas Zoltan J.
Published: December 2022

The rat mammary adenocarcinoma 13762A is weakly immunogenic in syngeneic hosts. Transplantation immunity is not developed after injection of irradiated tumor cells. Two cell types were isolated from the tumor: one grew slowly forming solid implants; the second grew rapidly in ascites form, even in allogeneic hosts. Spleen cells cytotoxic for a tissue culture derivative of the tumor were produced in animals after injection of either cell type indicating a common antigen.

The kinetics of tumor growth and production of cytotoxic spleen cells were compared in syngeneic and allogeneic animals. In vivo rejection of the tumor in allogeneic hosts did not correlate with the in vitro assay of spleen lymphocyte cytotoxicity. Furthermore, the cytotoxic response of spleen cells from syngeneic tumor-bearing hosts parallels that found in allogeneic hosts.
The Humoral Response of Mouse Spleen Cells to Two Types of Sheep Erythrocytes
Authors: McCarthy Margaret M. , Dutton Richard W.
Published: December 2022

It was previously shown that there is multigene control of the response of mouse spleen cells to two types of sheep erythrocytes (H and L). Discriminator strains of mice make a much higher response to extra antigens found only on H SRBC than to the shared antigens found on both types of erythrocyte. Non-discriminator strains respond only to the shared antigens, making a response as great as the discriminator response to the extra antigens. In this study we have shown that the response to the extra antigen(s) on H SRBC is under the genetic control of a gene(s) located in the VH region near or to the left of the A5a, α1,3-dextran and NP markers.
Inhibition of Tumor Cell Proliferation: A Second Role for Suppressor Cells?
Authors: Thomas David W. , Talmage David W. , Joberts Walden K.
Published: December 2022

An antimitotic factor for mouse tumor cells was isolated from the supernatant fluids of antigen-stimulated ovalbumin-immune mouse spleen cells. A similar antimitotic factor was obtained from the supernatant fluids of phytohemagglutinin-stimulated non-immune spleen cells. The inhibitor prevented the multiplication of mouse L929 fibroblast cells in vitro and the in vivo proliferation of Ehrlichascites tumor cells. Inhibition was not due to cytotoxic effects and the antimitotic effects were reversible. The antimitotic activity may be species-specific since the purified factor obtained from mouse spleen cells had no effect on human or monkey cell lines. This factor appears to be the same as the T lymphocyte-dependent suppressor for antibody production that we described previously. Several implications for the production of the suppressor in response to tumor cells that may be beneficial or harmful to the host are discussed.
Quantitation of Mouse Immunoglobulin Allotypes by a Modified Solid-Phase Radioimmune Assay
Authors: Bosma Melvin J. , Marks Robert , De Witt Carol L.
Published: December 2022

Allotype-reactive polystyrene tubes were made by first coating their inner surface with immunoglobulin (Ig) solutions containing the allotype of interest; this was followed by the application of anti-allotype antibody. The latter was covalently cross-linked to the adsorbed allotype antigen by means of glutaraldehyde to prevent the dissociation of antibody from the surface of the tube in subsequent steps (glutaraldehyde treatment did not affect the specificity of these tubes). Under conditions of antibody excess, allotypereactive tubes were treated next with 125I-labeled 7S Ig from standard solutions in order to estimate the concentration of Ig allotypes in these solutions. Then, to determine serum allotype concentrations, we used the standard solutions and the serum unknown to carry out competition experiments between labeled and unlabeled allotype antigen. Quantitation of serum Ig allotypes in this way featured a high signal to noise ratio (30 to 100), an error estimate of 3 to 7%, and sensitivity in the nanogram range.
Genetic Complementation of Histocompatibility-Linked Ir Genes in the Rat
Authors: Günther Eberhard , Rüde Erwin
Published: December 2022

Different inbred rat strains that are congenic with respect to the major histocompatibility system (H-1) show either a low, an intermediate, or a high immune response to the synthetic polypeptide (H,G)-A--L. Two out of four low responder H-1 haplotypes are found to complement intermediate responsiveness to high responsiveness. This effect is observed for the primary and secondary response and is detectable at the humoral and cellular level, i.e., antibody production and anamnestic response in vitro. Interallelic and intergenic complementation models are outlined. The ensuing implications for H-linked Ir gene control of immune responsiveness and for the structure of major histocompatibility systems are discussed.
Immunosuppression of Normal Lymphoid Cells by Serum from Mice Undergoing Chronic Graft-vs-Host Disease
Authors: McMaster Robert , Levy Julia G.
Published: December 2022

Spleen cells from F1 mice undergoing chronic graft-vs-host (GVH) reaction, induced by injection of parental cells, were shown to be immunosuppressed since their in vitro responses to the mitogens concanavalin A (Con A) and bacterial lipopolysaccharide (LPS) were substantially lower than control animals. Serum, from mice undergoing GVH, when cultured in vitro with normal spleen cells was immunosuppressive. The proliferative response to Con A and allogeneic cells of normal syngeneic, allogeneic, and parental spleen cells was 90% suppressed when serum from mice undergoing chronic GVH was added in comparison to the addition of serum from untreated F1 mice. Similarly, the in vitro antibody response to a T-dependent antigen was impaired; however, the antibody response to a T-independent antigen was not impaired. These results indicate that T cell functions are more sensitive than are B cell functions to immunosuppressive factors in the serum of mice undergoing GVH.
DNA Release As a Direct Measure of Microbial Killing
Authors: Friedlander Arthur M.
Published: December 2022

A new method for quantification of microbial killing is presented, based upon the assumption that release of DNA from an organism can be taken as direct evidence of cell death. The assay was applied to measurement of serum bactericidal activity. A “serum-sensitive” Escherichia coli, whose DNA was pre-labeled with [14C] thymidine, released 95 to 100% of its radioactivity upon exposure to human serum for 120 min. This was accompanied by a fall in viability with <0.1% of the bacteria surviving. Heating serum to 56°C for 30 min completely abolished both DNA release and killing. Normal serum did not release DNA from a “serum-resistant” Salmonella typhimurium whereas exposure to ampicillin caused both significant killing and DNA release. This assay is highly specific, sensitive, and rapid.
Effect on Tolerance Induction of the Mode of Attachment of the Hapten to the Carrier
Authors: Paley Robert S. , Leskowitz Sidney , Borel Yves
Published: December 2022

In this study we have investigated the role played by the mode of attachment of the hapten on autologous IgG in the induction of carrier-determined tolerance. Three conjugates of the dinitrophenyl hapten and mouse γ-globulin were prepared: 1) DNP-ɛ-lysine-MGG, 2) DNP-azo-MGG (which binds to histidine and tyrosine), 3) DNP-must-MGG (which binds to the carboxyl groups of glutamic or aspartic acid). DNP was also bound to the immunogenic carrier keyhole limpet hemocyanin (KLH) by the three different linkages. The three DNP mouse γ-globulin conjugates were used to induce tolerance and immediately thereafter the animals were immunized with various DNP-KLH conjugates. Antibody response was tested by a modification of the Jerne plaque assay. It was found that DNP-ɛ-lysine-MGG induced tolerance to both DNP-ɛ-lysine-KLH and to DNP-azo-KLH. In contrast, neither DNP-azo-MGG nor DNP-must-MGG induced tolerance to DNP-ɛ-lysine-KLH or to DNP-azo-KLH and DNP-must-KLH, respectively. Clearance studies showed no differences in the catabolic rate of the various hapten mouse γ-globulin conjugates as compared to unconjugated MGG. However, although DNP-must-MGG and DNP-azo-MGG were not tolerogenic, they were immunogenic.

In conclusion, this study demonstrated that not only the nature of the carrier but the mode of hapten binding to this carrier are critical for the induction of hapten specific tolerance.
Age-Related Changes in Cell-Mediated Immunity in BALB/C Mice
Authors: Walters Curla S. , Claman Henry N.
Published: December 2022

The effect of increasing age on various tests of cell-mediated immunity was investigated in BALB/c mice both in vitro and in vivo with four different assay systems. The following results were obtained. 1) In contact sensitivity to DNFB, old mice (age 60 to 80 weeks) showed no differences in sensitization when compared to young adult mice (age 8 to 12 weeks). (In contrast, old NZB/W mice showed impaired contact sensitization when compared with young NZB/W mice.) 2) Unlike the reaction in contact sensitivity, cells from old BALB/c mice were defective in eliciting a graft-vs-host reaction. This was true also when a partially purified population of T cells was transferred. 3) In the mixed lymphocyte reaction, cells from old mice were as efficient or better than cells from young adult BALB/c mice in responding to or stimulating allogeneic cells. 4) Responses to PHA and Con A (both T cell mitogens) were greatly reduced when old cells were cultured as compared with cells from young adult mice. Thus, we have found that within the same batch of mice, increasing age was associated with increased capabilities in some measures of cell-mediated immunologic function and decreased capabilities in other measures of the same.
Isolation of the Most Immunoreactive Antigens of Echinococcus Granulosus from Sheep Hydatid Fluid
Authors: Pozzuoli Roberto , Plantelli Mauro , Perucci Carlo , Arru Efisio , Musiani Piero
Published: December 2022

This paper describes a simplified procedure for obtaining purified Echinococcus granulosus antigens from sheep hydatid fluid by using affinity chromatography on concanavalin A-Sepharose. The presence of two “major” antigens (4 and 5) was confirmed. Antigen 5 was isolated by preparative polyacrylamide gel electrophoresis. Antigen 4, eluted by diffusion from the gel, was seen to be “contaminated” by antigen 5 and was isolated by using anti-5 Sepharose-linked serum. These two major antigens were then tested separately against the sera of hydatidosis patients by using very simple immunologic tests. The best results were obtained in passive hemagglutination with antigen 4. Antigen 4 is the most immunoreactive parasitic antigen; antibodies against it were found in the sera of all hydatidosis patients showing a positive reaction. Apart from the direct use of this antigen in serologic tests, it appears possible to standardize the most frequently used and commercially available antigenic materials by titrating this component.
Specifically Cytotoxic Human and Mouse Lymphood Cells Induced with Antibody or Antigen-Antibody Complexes
Authors: Saksela E. , Imir T. , Mäkelä O.
Published: December 2022

Human or mouse lymphoid cells could be “armed” with anti-NIP antibodies to become cytotoxic to NNP-conjugated fowl erythrocytes (NNP and NIP are closely related haptens). The arming factor was neutralized by a sufficient concentration of NIP-BSA (twice the concentration causing maximal precipitation) but low concentrations (e.g., 7% of the maximal precipitation concentration) increased the arming capacity.
Maintenance of Lymphocyte Surface Ig by Mitogen Stimulation in Vitro
Authors: Teodorescu Marius , Buchholz Donna M. , Dray Sheldon
Published: December 2022

In 4 to 24 hr cultures of rabbit lymphoid cells in medium supplemented with autologous serum, most B cells lost their surface Ig as assayed by rosette formation with anti-Ig antibody-coated erythrocytes. This loss was prevented by adding selected mitogens such as streptococcal mitogen (SM), lipopolysaccharide, and concanavalin A or by supplementing the medium with fetal calf serum. When SM was added at various times to the cultures (1, 2, 3, or 4 hr), it was effective in maintaining the approximate level of Ig-bearing cells present at the time of its addition but was ineffective in restoring the level of Ig-bearing cells present at the time the cultures were intiated. Very small, submitogenic doses of SM were sufficient to maintain the level of Ig-bearing cells. The data suggest that lymphocytes require continuous stimulation to maintain their surface receptors.
Oxisuran and Immune Reactions: Mediation of Oxisuran Action by the Adrenal Glands
Authors: van Dijk Hans , Bakker Irma A. , Testerink Jan , Bloksma Nanne , Willers Jan M.
Published: December 2022

Oxisuran was tested in a TAR system and for the DH on SRBC. Besides immunosuppressive properties in the TAR and DH also mentioned by other authors, stimulation was found after prolonged administration. No effect of oxisuran on the humoral response was observed. The action of oxisuran was suggested to be indirect, effected by stimulation of the PAG system. Adrenalectomy of mice made animals non-liable for oxisuran action measured in the TAR assay. Oxisuran caused an increase of the corticosterone plasma level with a peak after four daily injections. This was paralleled by a peak in adrenal gland weight and a minimum in thymus size. The reduction in these effects after five injections (tachyphylaxis) suggests that a feedback mechanism is involved. Comparison of thymus weights after oxisuran and corticosterone administration indicates that a daily dose of 100 mg oxisuran/kg corresponds with 1.5 mg corticosterone/kg. The effects of oxisuran in the TAR and on the DH and the feedback phenomenon could be imitated with 1.5 mg corticosterone/kg. The mononuclear phagocyte has been proposed as the main target cell of oxisuran action, mediated by the PAG system.
IgE-Induced Release of A Platelet-Activating Factor from Rabbit Lung
Authors: Kravis T. C. , Henson P. M.
Published: December 2022

Sensitized rabbit lung fragments release a platelet-activating factor (PAFL) after challenge with specific antigen or monospecific antibody to rabbit IgE. This release requires calcium and is less evident in lungs from rabbits producing IgG as well as IgE antibody. The PAFL released from lung stimulates the secretion of serotonin from washed rabbit platelets. PAFL is distinguishable from ADP or thrombin and has properties similar to PAF derived from basophils (PAFB). It is not, however, identical to PAFB since rabbit platelets specifically desensitized to PAF still respond by releasing serotonin if stimulated with PAFL.
Absolute Macrophage Dependency of T Lymphocyte Activation by Mitogens
Authors: Rosenstreich David L. , Farrar John J. , Dougherty Suanne
Published: December 2022

A T lymphocyte subpopulation that contains only 0.3% macropages and less than 2% B lymphocytes has been prepared from guinea pig lymph node cells by the use of two different types of adherence columns. This subpopulation does not proliferate in response to the mitogens Con A or PHA unless additional macrophages are added. The means by which macrophages restore T cell responsiveness to PHA has been investigated. Macrophages appear to function via two distinct mechanisms in this experimental situation. The first mechanism involves the binding of PHA to the macrophage followed by the “presentation” of the mitogen to the T lymphocyte in a manner that induces cell activation. This presentation function requires that the macrophage be viable and metabolicially active.

The second mechanism by which macrophages function is by the elaboration of a soluble factor or factors. The presence of these factors has been reliably and reproducibly demonstrated by using a double-chambered, Marbrook-type tissue culture vessel. This soluble factor can induce activation of T lymphocytes with surface bound PHA in the apparent absence of any form of macrophage presentation. In contrast, the function of this factor is clearly distinct from that of the reducing agent, 2-mercaptoethanol, (2-ME) since 2-ME does not enable this T cell subpopulation to be activated by mitogens.

On the basis of these observations, we propose that two distinct signals are required to activate this T lymphocyte subpopulation. One signal is delivered by the interaction of the mitogen with the T cell surface, and the second singal is delivered by a soluble factor(s) produced by macrophages. Whether all types of T lymphocytes require two signals to be activated, remains to be established.
Studies on the Mechanism of Lymphocyte-Mediated Cytolysis
Authors: Thorn Richard M. , Henney Christopher S.
Published: December 2022

The role of protein synthesis in the mechanism of T cell-mediated cytolysis has been re-investigated. Cytolytically active (C57BL/6 anti-DBA/2) spleen cells treated with pactamycin (10-7 M to 10-6 M) exhibited suppressed protein synthesis (100 ± 5%), but unimpeded lytic activity. Drug-treated effector cells, incubated for prolonged (up to 24 hr) periods of time in the presence and absence of antigen, showed no significant diminution of lytic activity although the incorporation of 3H-leucine into protein was totally ablated. Studies with emetine, another irreversible inhibitor of protein synthesis, gave identical results. These findings are difficult to reconcile with the hypothesis that effector T cells cause lysis via a soluble protein mediator.
In Vitro Generation of Suppressor Cell Activity: Suppression of in vitro Induction of Cell-Mediated Cytotoxicity
Authors: Hodes Richard J. , Hathcock Karen S.
Published: December 2022

It was observed that when normal mouse spleen cells were cultured alone in vitro (precultured) for 3 to 7 days, these cells lost the ability to generate cell-mediated cytotoxicity (CML) during subsequent in vitro sensitization with allogeneic spleen cells, trinitrophenyl (TNP)-modified syngeneic spleen cells, or syngeneic tumor cells. These precultured cells, which were themselves unable to generate CML, were also shown in mixing experiments to suppress, actively, the generation of CML by freshly explanted spleen cells. Suppression occurred at the sensitization phase of CML, and not at the effector level; supernatants from suppressive precultured cells were not suppressive. Suppression was totally abrogated by the treatment of spleen cells with a T cell-specific rabbit anti-mouse brain serum and complement (RαMB + C) either before or after preculturing, suggesting that a T cell was essential both to the generation of suppressor activity and to its expression. Suppressor activity was entirely absent in precultured nylon wool column-nonadherent spleen cells, a T cell-enriched population containing most of the RαMB + C-sensitive cells in the spleen. Precultured nylon column-adherent cells (T cell-depleted) did have suppressive activity, and a mixture of nylon-adherent and nylon-nonadherent cells was as suppressive after preculture as the precultured unseparated spleen. Moreover, the ability of nylon-adherent spleen cells to generate suppressive activity during preculturing was abrogated by treatment with RαMB + C. Thus, the “spontaneous” generation of CML-suppressive activity was dependent upon a limited subpopulation of splenic T cells isolated in the nylon columnadherent fraction. The relationship of these data to a previously described synergy between subpopulations of normal mouse spleen in the generation of CML is discussed, and the findings related to other suppressor systems described in the literature.
Enhancement of Reaginic and Hemagglutinating Antibody Production by an Extract of Bordetella Pertussis Containing Histamine Sensitizing Factor
Authors: Lehrer Samuel B. , Vaughan John H. , Tan Eng M.
Published: December 2022

The effect of an extract containing the histamine-sensitizing factor (HSF) of Bordetella pertussis on the immune response of mice to ovalbumin was investigated with respect to dose of antigen and adjuvant. Of particular interest was the enhancement of reaginic antibody production. In comparison to the Al(OH)3 induced production of reaginic antibody where low doses of antigen and adjuvant yield high titers of reagin, the HSF extract demonstrated optimal adjuvant activity at high doses of both adjuvant and antigen. The reaginic antibody response was maximal usually by 2 to 3 weeks post-immunization and persisted for long periods of time. The hemagglutinating antibody response was maximal at 8 to 10 weeks post-immunization. The initial treatment of mice with HSF extract plus antigen resulted in the production of memory cells since a subsequent immunization with ovalbumin alone evoked a secondary reaginic antibody response. These observations may have implications in clinical allergy since substances similar to the pertussis factor might be produced by other microbial organisms and these substances could modulate the immunologic response of individuals to common allergens.
Cellular Receptors for Lymphotoxin: Correlation of Binding and Cytotoxicity in Sensitive and Resistant Target Cells
Authors: Tsoukas Constantine D. , Rosenau Werner , Baxter John D.
Published: December 2022

The binding of radioactively labeled lymphotoxin (LT) to both lymphotoxin-sensitive and -resistant cell clones was examined. The sensitive clone had a low-capacity, high-affinity (“specific”) binding component, the curve of which closely followed the cytotoxicity curve of the lymphocyte mediator. The capacity of this binding component was calculated to be about 600 molecules of LT/cell. In addition, there was a low-affinity, high-capacity (“nonspecific”) binding component. In striking contrast, the high-affinity, low-capacity (“specific”) component was absent or greatly diminished from the resistant clone, whereas the low-affinity, high-capacity (“nonspecific”) component was present at a similar level as in the sensitive cells. These binding characteristics closely resemble those observed by us and other investigators working with a variety of steroid hormones in steroid-sensitive and -resistant cell lines.
Antigenic Determinants of Adenovirus Capsids
Authors: Willcox Nicholas , Mautner Vivien
Published: December 2022

Evidence is presented that the type-specific antibody to the adenovirus hexon is not simply the antibody with the highest avidity for cross-reactive determinants, but is a distinct, minority population that recognizes separate determinants. To quantify it, we have developed an inhibition method with radio-immunoprecipitation (RIP) as a sensitive assay for the type-specific antibody that remains after all the excess of cross-reactive antibody has been blocked by heterologous antigen.

During the primary response, 0.1 to 1% of antibody to types 2 or 5 hexon is type-specific, but after boosting, this population may reach 10 to 20%. Antibody to fiber is more than 70% type-specific during primary and secondary responses. The cross-reacting antibody can be removed on immunoabsorbent columns without affecting the virus neutralization titer of the serum.
Cell-Mediated Immunity to Friend Virus-Induced Leukemia
Authors: Ting Chou-Chik , Kirchner Holger , Rodrigues Dennis , Park Joseph Y. , Herberman Ronald B.
Published: December 2022

By employing the 125IUdR release cytotoxicity assay, we have been able to measure the primary and secondary cell-mediated cytotoxic response of C57BL/6 mice to FBL-3 cells, a syngeneic Friend virus-induced leukemia. It was found that the secondary cell-mediated cytotoxic response occurred more rapidly after challenge (within 3 days) than the primary response, and the levels of reactivity were considerably higher. As in the primary response, the secondary cytotoxic reactivity of spleen cells was T cell dependent, being eliminated by pretreatment with anti-θ antibody plus complement. However, the secondary reactivity of pertioneal exudate (PE) cells was not entirely T-cell dependent. The specificity of the secondary cytotoxic response was analyzed by primary or secondary immunization with various tumor cells and by testing of cytotoxic lymphocytes against a variety of target cells. When spleen cells were used for testing, only tumor cells induced by Friend, Moloney, or Rauscher (FMR) leukemia viruses could produce secondary cell-mediated cytotoxic responses against FBL-3 cells. This correlated well with the specificity observed in the in vivo tumor transplantation protection studies. Similarly, spleen cells immune to FBL-3 had appreciable cytotoxicity against tumor cells induced by FMR viruses. The FBL-3 immune mice also gave significant protection against the challenge of FMR leukemias. When PE cells were used for testing, they gave a broader spectrum of reactivity. These cells usually gave higher levels of cytotoxicity against tumor cells induced by FMR viruses, but also gave less, but appreciable, cytotoxicity against non-FMR tumors. The latter reactivity might be related to the antigens induced by the murine endogenous type C viruses.
Kinetics of the Antibody Response to Type III Pneumococcal Polysaccharide (SSS-III)
Authors: Jones Jeffrey M. , Amsbaugh Diana F. , Prescott Benjamin
Published: December 2022

A simple method was described for the preparation of 125I-labeled type III pneumococcal polysaccharide (SSS-III*) with a high specific radioactivity which retained the physical and immunologic properties of native SSS-III. SSS-III* was used to study the serum and tissue levels of antigen, as well as its excretion, after i.p. injection. When an optimally immunogenic dose (0.5 µg) of antigen was given, τ;90% of the injected antigen was excreted during the first 3 days after injection; however, after day 3, the SSS-III* which remained in each mouse was firmly bound to various tissues, and <5 ng SSS-III* was released into the circulation daily. SSS-III* was also used in a Farr test to measure serum antibody levels; the kinetics for the appearance of PFC/spleen and serum antibody levels were measured at 24-hr intervals after immunization with 0.5 µg of antigen. Maximum PFC/spleen were observed on day 4 after immunization whereas the peak serum antibody level was seen on day 5. The decay of serum antibody level from its maximum value was much slower than that of the PFC/spleen. The data describing the distribution of SSS-III in vivo and the measurement of serum antibody levels indicated that treadmill neutralization was not a factor in determining the serum antibody levels after immunization with an optimally immunogenic dose of SSS-III.
Purification of the Sixth and Seventh Component of Human Complement without Loss of Hemolytic Activity
Authors: Podack Eckhard R. , Kolb William P. , Müller-Eberhard Hans J.
Published: December 2022

Procedures for the isolation of the human complement proteins C6 and C7 have been described. These procedures allow isolation of the two proteins without any loss of hemolytic activity. Apparent activity gains of 160% and 140% were observed for C6 and C7, respectively, when the activity of the isolated proteins was compared with their activity in serum. The recovery of C6 was 3.5 to 11% and that of C7 was 7 to 13% of the amount present in serum. C6 has a m.w. of 128,000 and an electrophoretic mobility at pH 8.6 of -2.6 × 10-5 cm2 s-1 v-1. C7 has a m.w. of 121,000 and an identical electrophoretic mobility. With 3 × 107 assay cells, 63% hemolysis was achieved with 1 ng of C6 and 3.8 ng C7. On polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and after reduction with mercaptoethanol, C6 and C7 behaved as single polypeptide chain proteins.
Identification of a Macrophage-Specific Cell Surface Antigen
Authors: Stinnett J. Dwight , Kaplan Alan M. , Morahan Page S.
Published: December 2022

A mouse-specific macrophage antigen (MSMA) was identified in NP-40 extracts of 125I-radiolabeled mouse peritoneal macrophages by using a rabbit anti-mouse macrophage serum (AMS) and SDS-polyacrylamide gel electrophoresis. The antigen was shown to have a m.w. of 83,000 daltons and was present on both normal and “activated” peritoneal macrophages. MSMA was also present on syngeneic adherent spleen cells, allogeneic peritoneal macrophages, a mouse macrophage cell line (P388D1), and exhibited some cross-reactivity with peritoneal macrophages from closely related species (rats and hamsters). MSMA was not present on nonadherent peritoneal exudate cells, spleen cells, erythrocytes, thymocytes, or bone marrow cells. Extensive absorptions of AMS with thy - mocytes and erythrocytes from mice were necessary to remove other antibodies that reacted with other mouse membrane antigens. An antiserum directed against a specific membrane antigen has great potential in elucidating structure-function relationships with regard to a number of macrophage activities.
Spontaneous Cytotoxicity (SCMC) of Normal Human Lymphocytes against a Human Melanoma Cell Line: A Phenomenon Due to a Lymphotoxin-Like Mediator
Authors: Peter Hans H. , Eife Rudolf F. , Kalden Joachim R.
Published: December 2022

Spontaneous cell-mediated cytotoxicity (SCMC) of normal human lymphocytes against various allogenic tumor cell lines has recently been identified as a non-T lymphocyte function. In this study evidence is presented that SCMC against a human melanoma cell line (IGR3) involves a nonspecific lymphotoxin-like mediator(s) (LT), which is rapidly produced by lymphocytes that are retained on IgG-anti-IgG columns. LT was shown to inhibit in 48-hr cultures the DNA synthesis of growing IGR3 and HeLa cell monolayers. Furthermore, in short term 51Cr-release assays using IGR3 target cells, LT increased strongly the SCMC of normal allogeneic lymphocytes, although it exhibited little cytotoxicity by itself in this assay. LT was detectable in cell-free supernatants harvested after 6 hr from co-cultures of IGR3 melanoma cells and normal effector lymphocytes, but not in supernatants from melanoma cells alone or lymphocytes alone. Lymphocyte preparations that had been passed through IgG-anti-IgG columns had lost the capacity to generate LT and were poor effectors in SCMC. However, in the presence of LT, a small proportion of null cells, which pass through IgG-anti-IgG columns, was capable of inducing strong SCMC. Absorption of the supernatants, containing LT activity, with an insolubilized rabbit-anti-human IgG antiserum did not remove the mediator, suggesting that it is not an immunoglobulin.
The Role of the Capsular Polysaccharide in the Activation of the Alternative Pathway by the Pneumococcus
Authors: Winkelstein Jerry A. , Bocchini Joseph A. , Schiffman Gerald
Published: December 2022

Previous studies have shown that when pneumococci are incubated in normal, nonimmune serum, they activate the alternative pathway and opsonically active C3b is fixed to the surface of the organism. Other studies have demonstrated that C3-dependent opsonization via the alternative pathway plays a significant role in the nonimmune host's defense against the pneumococcus. The present studies concern the role of the capsular polysaccharide in initiating the activation of the alternative pathway by the pneumococcus.

Some pneumococcal capsular polysaccharide types, but not all, are able to activate the alternative pathway. Soluble purified capsular polysaccharide types 1, 4 and 25 activate the alternative pathway, whereas types 2, 3, 14, and 19 do not. Since the capsular polysaccharides exist in their native form attached to the pneumococcal surface, selected capsular polysaccharides were also tested for their ability to activate the alternative pathway when attached to a particulate carrier, sheep erythrocytes. Capsular polysaccharide types 2 and 3 failed to activate the alternative pathway when attached to sheep erythrocytes, paralleling the results obtained when these capsular polysaccharides were in solution. In contrast, the type 25 capsular polysaccharide not only activated the alternative pathway when attached to sheep erythrocytes, as it had when in solution, but is also initiated alternative pathway-mediated lysis of the erythrocytes.

The capsular polysaccharide is not required for the activation of the alternative pathway by the pneumococcus. Although all types of encapsulated pneumococci are able to activate the alternative pathway, not all the purified capsular polysaccharide types are able to do so. In addition, a nonencapsulated pneumococcus, derived originally from a type 2 organism, activates the alternative pathway as well as a fully encapsulated type 2 pneumococcus.
Isolation and Characterization of Rabbit Antibodies Directed against Group a Allotypic Determinants
Authors: Aasted Bent , Sogn John A. , Kindt Thomas J.
Published: December 2022

The L chains of rabbit antibodies directed against group a allotypes exhibited in many instances a high degree of homogeneity as measured by L chain banding patterns in alkaline urea polyacrylamide gels. Amino terminal sequence analyses were carried out on six L chain preparations from antibodies isolated from individual antisera or from pools of antisera. The same major amino terminal sequence, Ala-Val-Val-Met, was observed for each of these preparations indicating that anti-allotype antibodies preferentially select L chains from a single subgroup.

The antiallotype antibodies were isolated from antisera by elution from IgG immunoadsorbent columns in yields ranging from 0.3 to 1 mg/ml antibody. The specificity of the isolated antibodies was demonstrated by radioimmune assays. Certain fractions were contaminated with a protein that had properties similar to rabbit serum albumin. This contamination was minimized by preadsorption of the antisera. The antibodies were primarily of the IgG class as shown by immunoelectrophoresis and by m.w. of the H and L chains on SDS gels.
The Effect of Thiols on the Immunologic Release of Slow Reacting Substance of Anaphylaxis
Authors: Orange Robert P. , Moore Elizabeth G.
Published: December 2022

In the presence of L-cysteine, a selective and marked enhancement of the in vitro, immunologic release of slow reacting substance of anaphylaxis (SRS-A) from human peripheral leukocytes, sensitized monkey lung fragments, and sensitized guinea pig lung fragments was observed. In the rat, cysteine, but not sodium sulfide, enhanced the calcium ionophore (A23187)-induced release of SRS-A in vitro from mixed rat peritoneal cells and in vivo from the rat peritoneal cavity. Pretreatment of rats with cysteine also enhanced the IgGa- and anti-rat IgE-mediated release of SRS-A in vivo in the rat. These studies indicate a common biochemical mechanism involved in the formation and release of SRS-A from these different tissues and cells and further confirm the observation that the rat mast cell is not a major source of SRS-A in the rat.