bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunogenic Properties of Modified Antigen E
Authors: Takatsu Kiyoshi , Ishizaka Kimishige , King Te Piao
Published: December 2022

It has been shown that regweed antigen E loses its major antigenic determinants after denaturation in 8 M urea, but urea-denatured (UD) antigen and an α-polypeptide chain isolated from the denatured molecules are capable of priming mouse T cells specific for native antigen. Weekly injections of 10μg UD antigen or α-chain into antigen E-primed antimals depressed the ongoing IgE antibody response, whereas injections of the same dose of antigen E failed to depress the antibody response. It was found by adoptive transfer experiments that helper activity of antigen E-primed splenic T cells was depressed by the treatment of the donors with either modified antigen or native antigen E. The same treatment of antigen E-primed animals depressed the DNA synthetic response of their splenic T cells to antigen E. The treatment of antigen E-primed animals with UD antigen resulted in a decrease of antigen E-specific IgE-B cells and IgG-B cells in their spleen, whereas the treatment with native antigen expanded the B cell populations. In view of the results obtained in the mouse, cellular basis for the immunologic effects of hyposensitization treatment is discussed.
Immunologic Functions of Isolated Human Lymphocyte Subpopulations
Authors: Chess Leonard , Levine Herbert , MacDermott Richard P. , Schlossman Stuart F.
Published: December 2022

Sephadex G-200 anti-human Fab column chromatography and rosette depletion techniques were used to isolate three distinct subpopulations of human lymphocytes: 1) T cells which are surface Ig negative and E rosette positive, 2) B cells which are surface Ig positive and E rosette negative, and 3) a “Null” cell population which is both surface Ig negative and E rosette negative. All populations were analyzed for their capacity to develop surface Ig and synthesize Ig in vitro. Greater than 50% of cells in the Null cell population developed surface Ig by day 3 of cell culture. Furthermore, in vitro, the Ig content of the Null cell population, as well as their capacity to secrete Ig in culture, becomes comparable to that produced by B cells. In contrast, cultured T cells neither develop surface Ig nor secrete Ig in culture. These data strongly support the idea that the Null population contains a subset of Ig-producing B cells.
Cell-Mediated Immune Responses in Vitro
Authors: Peavy Duane L. , Pierce Carl W.
Published: December 2022

The conditions for generation of simultaneous and independent cytotoxic lymphocyte (CL) responses to each of two sets of alloantigens of limited cross-reactivity by mouse spleen cells in vitro have been investigated. Responder spleen cells were incubated with mitomycin C-treated C57BL/6 (H-2b) or DBA/2 (H-2d) stimulator spleen cells and day 5 CL responses were assayed with 51Cr-labeled EL-4 leukemia (H-2b) and P815 mastocytoma (H-2d) as target cells. Spleen cells from mice of the various H-2 haplotypes tested differed greatly in their ability to develop specific CL responses against alloantigens on the stimulator spleen cells and in the degree of cross-reactive cytotoxic activity against target cells bearing alloantigens not present on the stimulator spleen cells. In contrast to the other strains examined, DBA/1 (H-2q) spleen cells developed specific CL responses to either H-2b or H-2d alloantigens without exhibiting significant cross-reactive activity on the inappropriate target cell. The CL responses to H-2b and H-2d alloantigens by DBA/1 spleen cells were comparable in magnitude and had similar stimulator cell-dose requirements. Further, DBA/1 spleen cells developed CL responses of normal magnitude simultaneously against both target cells when incubated with both mitomycin C-treated C57BL/6 and DBA/2 stimulator cells.
Cell-Mediated Immune Responses in Vitro
Authors: Peavy Duane L. , Pierce Carl W.
Published: December 2022

The role of cellular proliferation in the development of cytotoxic lymphocyte (CL) responses in one-way mixed lymphocyte reactions was investigated by using tritiated thymidine of high specific activity to kill proliferating cells. To develop maximum CL responses, responding lymphoid cells must proliferate for approximately 72 hr; thereafter, precursors of CL appear to differentiate into active CL without further proliferation. Different alloantigen-sensitive precursor cell populations participate in the CL responses to each of two sets of stimulating alloantigens. When cells responding to one set of alloantigens were selectively destroyed after incorporating the hot thymidine, the surviving cells retained the capacity to develop a normal CL response to the second set of alloantigens.
Release of Bound Immunoglobulin from SSPE Brain by Acid Elution
Authors: Weil Marvin L. , Leiva William A. , Heiner Douglas C. , Tourtellotte Wallace W.
Published: December 2022

SSPE brain homogenate extracted at pH 7.4 yields immunoglobulin with a 4- to 5-fold greater hemagglutination inhibition activity per microgram of IgG than serum from the same patient. Serial washing of the homogenate results in a low level steady-state release of IgG. Elution of the washed sediment with pH 2.5, 0.1 M glycine buffer results in a 2- to 3-fold increase in recovery of hemagglutination inhibition activity with a greater hemagglutination inhibition activity per milligram of IgG than the IgG recovered by phosphate-saline extraction at pH 7.4.
Antigen Suppression of the in Vitro Development of Plaque-Forming Cells to Autologous Erythrocyte Antigens
Authors: Lord Edith M. , Dutton Richard W.
Published: December 2022

Peritoneal cell cultures from NZB and C57BL/6 mice develop large numbers of PFC directed against antigens present on bromelain-treated isologous erythrocytes. The development of these autoimmune PFC can be suppressed by the addition of small numbers of BrMRBC at the start of the culture period. The possibility that the development of the plaques is prevented by the presence of antigen in vivo is discussed.
Generation of Cytotoxic T Lymphocytes in Vitro
Authors: Fitch Frank W. , Engers Howard D. , MacDonald H. Robson , Cerottini Jean-Charles , Brunner K. Theodor
Published: December 2022

Reexposure of day 14 murine mixed leukocyte culture (MLC) populations to the original irradiated allogeneic stimulating spleen cells has previously been found to result in the rapid generation of cytolytic T lymphocytes (CTL) associated with a net increase in cultured cell number. Under the experimental conditions used, day 5 MLC cells appeared unable to respond to the allogeneic stimulus. In order to characterize further the development of the potential for anamnestic reactivity during the course of MLC, C57BL/6 spleen cells were incubated with irradiated (1000 rads) DBA/2 spleen cells (primary MLC) for up to 3 weeks. At various time intervals after the onset of the primary MLC, the surviving cells were collected and reexposed, at varying cell concentrations, to irradiated DBA/2 spleen cells (secondary MLC). At daily intervals thereafter, CTL activity was assessed using a quantitative 51Cr-release assay system.

A paradoxic effect of responding cell concentration on generation of CTL activity was observed; relatively greater increase in CTL activity was observed as the concentration of responding cells was decreased over a 100-fold range. This effect was more pronounced with responding cells reexposed to antigen after primary MLC for 20 days, but was observed even with normal cells. The apparent unresponsiveness of day 5 MLC cells to alloantigen restimulation could be overcome by simple dilution of responding cells. Cytotoxic activity at the time of restimulation with antigen seems to be a major factor determining the magnitude of the secondary response. Since intact cells bearing alloantigens are required for the generation of CTL in MLC, residual cytotoxic cells reduce the effective antigenic stimulus by destroying stimulating cells. This effect of concentration of responding cells on generation of CTL in MLC complicates interpretation of experiments investigating the role of “inhibitor” and “helper” cell in cell-mediated immune responses occurring in vitro.

Under optimal conditions, the highest CTL activity and the largest increase in total cell number was observed 4 days after restimulation of day 10 MLC cells. On a per cell basis, the lytic activity was up to 4 times greater than that observed at the peak of a primary response, and the number of viable cells recovered was nearly 20 times higher than that at the onset. Such secondary MLC are thus a convenient source of lymphoid cells selected primarily on the basis of proliferation induced by alloantigens.
Specific Tumor Immunity Induced with Soluble Materials: Restricted Range of Antigen Dose and of Challenge Tumor Load for Immunoprotection
Authors: Pellis Neal R. , Kahan Barry D.
Published: December 2022

Immunogenic materials were solubilized from two methylcholanthrene-induced fibrosarcomas using 3 M KCl and were assessed for antigen activity in an immunoprotection assay. Mice which had been pretreated with a single injection of extract (0.1 to 2.0 mg protein) were challenged 10 days later with 103, 104, or 105 tumor cells. Optimal protection, as evidenced by survival, tumor incidence, and prolonged latent period before neoplastic outgrowth, was found after pretreatment with 0.5 mg of crude KCl extract and challenge with 104 tumor cells. These results further reinforce the weak nature of tumor-specific antigens as immunogens and show that only in very restricted circumstances can solubilized preparations administered without adjuvant induce immunoprotection in syngeneic hosts.
Mouse Mononuclear Cell Chemotaxis
Authors: Boumsell Laurence , Meltzer Monte S.
Published: December 2022

The chemotactic responses of peripheral blood monocytes and peritoneal macrophages to lymphocyte- and complement-derived stimuli were studied in vitro. Monocyte and macrophage chemotactic responses to these stimuli were different in both dose-response and time course. These differences provide a useful functional marker between these two cell populations.
C3 Nephritic Factor (C3NeF): Stabilization of Fluid Phase and Cell-Bound Alternative Pathway Convertase
Authors: Daha Mohamed R. , Fearon Douglas T. , Austen K. Frank
Published: December 2022

C3 nephritic factor (C3NeF) defined by the capacity of nephritic serum and its fractions to initiate loss of the B antigen of C3 in normal serum was purified from the serum of three different donors and shown to function by stabilization of membrane-bound and fluid phase alternative pathway C3 convertase. C3NeF converts cell-bound C3B̅ sites in a dose-related manner to C3B̅(NeF) sites, which exhibit and approximate 10-fold increase in half-life. The linear relationship between the C3NeF input and the residual hemolytic sites on EAC43B present after incubation for 20 min at 30°C, during which labile C3B̅ sites, have decayed, indicates that the number of residual C3B̅ sites is directly related to the dose of C3NeF. The capacity of C3NeF to stabilize the C3B̅ convertase in a temperature- and dose-dependent manner, which is independent of binding or consumption of C3NeF, in a fluid phase reaction mixture of 125I-B, 131I-C3, and D̄ permits isolation of a 10S complex containing radiolabeled C3 and B and exhibiting C3 convertase activity on an exogenous C3 source. Thus, the stabilizing effect of C3NeF is not limited to membrane-bound C3B̅ but is also sufficient to permit recovery of a fluid phase C3 convertase formed during the interaction of C3, B, and D̄.
Experimental Autoimmune Orchitis: in Vitro Induction of an Autoimmune Disease
Authors: Wekerle H. , Begemann M.
Published: December 2022

Experimental autoimmune orchitis (EAO) can be induced in vitro. Normal lymph node lymphocytes cultured with autologous dissociated testis cells form rosette-like aggregates and later undergo blast transformation and proliferation. These stimulated lymphocytes cause in vivo EAO lesions, when injected into syngeneic recipients. Moreover, their autoimmune reactivity can be monitored by an in vitro cytostasis assay. Density gradient analysis of the early lymphocyte-testis cultures reveals that the autoimmune reactive lymphocytes are enriched in the rosette populations. It therefore appears that testicular self-antigens are recognized by clonally preformed autologous lymphocytes.
C1q Inhibition of the Interaction of Collagen with Human Platelets
Authors: Cazenave J. -P. , Assimeh S. N. , Painter R. H. , Packham M. A. , Mustard J. F.
Published: December 2022

Human C1q, isolated in pure state after affinity chromatography on IgG-Sepharose, inhibited collagen-induced aggregation and release of 14C-Serotonin from prelabeled human platelets. Platelet aggregation induced by ADP or thrombin was not inhibited by C1q. Also, the adherence of platelets to glass surfaces was significantly diminished by C1q. In contrast, aggregated C1q mimicked the effect of collagen in causing platelet aggregation and release of serotonin. It appears that monomeric C1q, which has structural similarities to collagen competes with collagen for specific sites on the platelet surface.
A Properdin System Intermediate Formed by Zymosan and Serum at 0°C
Authors: Yukiyama Yasushi , Lew Frances T. , Waks Harriet S. , Osler Abraham G.
Published: December 2022

The interaction of guinea pig or rat serum with Z at 0°C leads to the formation of a properdin pathway intermediate, ZPI, whose activity is assessed by its capacity to deplete the titer of cobra venom inducible lysis in diluted rat serum. The formation of ZPI does not require C1 or C2 and is not diminished by prior absorption of the serum with Z. In contrast, removal of P suppresses ZPI formation. Both Ca++ and Mg++ are essential cofactors for the formation of this intermediate whose function is abrogated in the presence of anti-C3, anti-P, but not by anti-Factor B. Since C4-deficient guinea pig serum is also effective in ZPI formation, the data suggest that ZPI is an alternative pathway intermediate containing both P and C3.
The Effect of Neonatal Rat Graft-vs-Host Disease (GVHD) on Fc Receptor Lymphocytes
Authors: Clancy John , Tønder Olav , Boettcher Carol E.
Published: December 2022

The level of Fc receptor rosette-forming lymphocytes (Fc-RFL) was examined in spleen and lymph node cell suspensions from neonatal DA and BN rats inoculated within 24 hr of birth with either allogeneic L (experimental) or syngeneic (control) lymphoid cells. In addition, these levels were compared to fetal and neonatal animals that received no injection. The indicator cells (EA) were sheep erythrocytes sensitized with one-half the concentration of the highest dilution of rabbit anti-sheep erythrocyte IgG(A) which agglutinated an equal amount of a 1% suspension of E. Care was taken to exclude scoring macrophages by injecting colloidal carbon at least 1 hr before killing the test animals. The spleen of 19-day DA fetal rats exhibited a level of 19.3% Fc-RFL, similar to that of animals having received adult syngeneic cells at birth (40.0%) by day 7. Thereafter the level of Fc-RFL did not vary appreciably between these two groups. However, as early as 2 days after inoculation there was a significantly greater number of Fc-RFL in the spleen of experimental DA neonates compared to controls. The lymph nodes of experimental animals did not exhibit a significantly greater number of Fc-RFL until day 6 with both tissue compartments peaking at day 10 and remaining significantly higher than controls until death. In neonatal BN animals significantly higher levels of Fc-RFL in experimental animals were not evident as early and peaked later (day 12) in both tissue compartments but again these differences remained until dealth. Cytotoxic alloantisera demonstrated that on days 6, 10, and 12 most, if not all, of the Fc-RFL were host in origion in both DA and BN GVHD, with a very significant host plasma cell response in such GVHD animals. One-micron tissue section revealed the presence of a great number of plasma cells especially prominent in the medulla of lymph nodes with the cortex of lymph nodes and white pulp of the spleen markedly depleted of lymphocytes indicative of cytotoxicity.
Age-Dependent Changes in Sensitivity to Antigen: Its Relationship with the Antigen Processing System
Authors: Carter Brian G. , Rector Edward S.
Published: December 2022

The relationship between the antigen processing or accessory cell system and the maturation, with age, of antibody-producing capability was investigated in the mouse. This was done by analyzing the antibody responses given by immunocompetent cells from neonatal and from adult mice in an identical antigen-processing system environment. Specifically, 4 × 107 normal spleen cells from either 12-day-old or adult mice were challenged with varying numbers of SRC in adult irradiated syngeneic recipients. The subsequent IgM and IgG2a PFC responses for both age groups were analyzed in terms of antigen dose-antibody response curves. Analyses of these curves indicate: 1) the dose of antigen required to elicit the optimal antibody response is essentially identical for both age groups and (2) the bandwidths obtained using neonatal donors are significantly narrower than those obtained with adult donors. Whereas, intact neonatal and adult mice exhibit differences in antigen optimal doses, these differences are eliminated when the immunocompetent cells are stimulated in the presence of identical antigenprocessing systems. It is concluded that maturation of the antigen-processing system results in an increased sensitivity to antigen. Examination of the bandwidths of the dose-response curves revealed that immunocompetent cells from the young mice, either in situ or in adult irradiated recipients exhibited narrower bandwidths than did their adult counterparts. Thus, the increase in bandwidth observed with age is attributed to changes in the population of immunocompetent cells—perhaps a reflection of increased diversity, and is not due to the antigenprocessing system.

Quantitation of the antigen-processing function using accessory cell-depleted and partially restored mice indicated that when IgG responses were compared a higher frequency of accessory cells was demonstrated in adult spleens as opposed to neonatal spleens.
A Thermostable Antigen Characteristic for Carcinogen-Induced Rat Intestinal Tumors
Authors: Abeyounis C. John , Milgrom Felix
Published: December 2022

Precipitation tests using properly absorbed rabbit antisera revealed that PCA extracts of DMH-induced rat jejunal and colonic adenocarcinomas contain an antigen that is not detectable in extracts of normal rat tissue or other rat tumors. This rat tumor antigen was detected in extracts of normal rat colon by employing the much more sensitive procedure of tanned cell hemagglutination inhibition, however, the concentration of the antigen in tumor was about 250-fold higher than in normal colon.

Similar to human CEA, the rat tumor antigen was found to be heat stable, contained glycoprotein, migrated as a β-globulin, and appeared at a high concentration in fetal intestines, however, the rat tumor antigen was serologically distinct from human CEA.
Regulation of Helper Cell Activity by Specifically Adsorbable T Lymphocytes
Authors: Rubin Bent
Published: December 2022

Mice immunized with soluble proteins such as human serum albumin (HSA) or ovalbumin (OA) develop in their spleens antigen-specific T and B lymphocytes. These populations of lymphocytes can be separated from each other by different means; e.g., treatment with anti-θ antiserum and complement removes selectively T lymphocytes, whereas passage through glass bead columns coated with mouse immunoglobulin (Ig): anti-Ig complexes creates a relatively pure population of T lymphocytes. During the course of such separation studies it was observed that the helper capacity of HSA (or OA) immune mouse spleen cells after Ig:anti-Ig column passage frequently was higher than expected from the enrichment in θ-positive cells. In addition, after adsorption onto antigen coated Bio-Gel beads this effect was even more pronounced, i.e., and increase in the relative helper capacity of about 3 or 4 times compared with an increase in the content of θ-positive cells from about 30% to 40 to 50% after adsorption.

The present results will demonstrate that the increased helper capacity was a specific phenomenon which was regulated by θ-positive cells. The regulatory cells specifically adsorbed onto antigen-coated Bio-Gel beads have not been successfully eluted by EDTA or excess-free antigen so far, and they were still adsorbed after pre-incubation with anti-Ig antibodies under conditions where specific B lymphocyte adsorption was almost prevented.
Regulation of the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Morse Herbert C. , Prescott Benjamin , Cross Sue S. , Stashak Philip W. , Baker Phillip J.
Published: December 2022

Mice of different ages were evaluated with respect to their ability to give a plaque-forming cell (PFC) response to Type III pneumococcal polysaccharide (SSS-III), as well as the degree of amplifier and suppressor thymus-derived (T) cell activity present. Although the magnitude of the PFC response to an optimally immunogenic dose of SSS-III for 2- and 3-week old mice was only 7% and 14%, respectively, of that produced by adult (8-week old) mice, values comparable to those of adult animals were attained by 4 weeks of age; no significant changes in the ability to respond to SSS-III occurred thereafter. Amplifier T cell activity, which was minimal at 2 to 4 weeks of age, matured slowly and did not reach a maximum until 8 to 10 weeks of age. By contrast, suppressor T cell activity appeared to be fully developed at least as early as 2 weeks of age; here, the inhibitory effects produced could be abrogated by depletion of T cells, indicating that the unresponsiveness induced by such cells does not result in the depletion or irreversible inactivation of B cells capable of responding to SSS-III. These findings suggest that the inhibitory effects of suppressor T cells are predominant in young mice and that such cells may play an important role in determining the ease with which unresponsiveness is induced in neonates, and in the prevention of autoimmune disease. Also, studies conducted with adult-thymectomized mice showed that both amplifier and suppressor T cells, once seeded to the periphery, are stable and do not depend upon the presence of intact thymus for the expression or renewal of their activity.
Diversity of the Antibody Response to the Different Antigenic Determinants on the Hemagglutinin Subunits of Influenza Viruses
Authors: Laver W. Graeme , Downie Jean C. , Webster Robert G.
Published: December 2022

Sera from rabbits hyperimmunized with hemagglutinin (HA) subunits isolated from the A/Port Chalmers/73 (H3N2) strain of influenza virus showed great differences in their cross-reactions with different strains of influenza virus. In hemagglutination-inhibition tests, some sera reacted to about the same titer with A/Port Chalmers/73 and A/Hong Kong/68 viruses, suggesting that these two strains were very closely related. Other sera, which reacted to high titer with A/Port Chalmers/73 virus, had only a low titer with the Hong Kong/68 strain, suggesting that the two viruses were distantly related.

Evidence suggested that these diverse cross-reactions were due to widely different ratios, in the different sera, of antibodies to the “common” and the “specific” antigenic determinants on the HA subunits. Thus, some rabbits gave a stronger response to the “common” determinants than to the “specific”, whereas in others, the reverse seemed to be the case.

Sera from human volunteers injected with A/Port Chalmers/73 inactivated or subunit influenza virus vaccines, or from people infected with Port Chalmers/73 virus, contained, in most cases, antibodies predominantly to the “common” antigenic determinants on the HA subunits. These sera reacted to higher titer with Hong Kong/68 virus than with the Port Chalmers/73 strain. Absorption of these sera with Hong Kong/68 virus totally removed all detectable antibody, suggesting that they contained no antibody to the “specific” determinants on Port Chalmers/73 HA. Paradoxically, absorption of the sera with Port Chalmers virus did not remove all antibodies, suggesting that the sera contained antibodies to the “specific” determinants on Hong Kong/68 HA.
A Microtiter Solid-Phase Radioimmunoassay for Hepatitis a Antigen and Antibody
Authors: Purcell Robert H. , Wong Doris C. , Moritsugu Yasuo , Dienstag Jules L. , Routenberg John A. , Boggs Joseph D.
Published: December 2022

A microtiter solid phase radioimmunoassay for hepatitis A antigen (HA Ag) and antibody (anti-HA) was developed. The test was more sensitive than immune adherence hemagglutination for detecting HA Ag and almost as sensitive for detecting anti-HA. The specificity and sensitivity of reagents were examined and optimum conditions for the test were determined. Radioimmunoassay, immune adherence hemagglutination, and immune electron microscopy were compared for detecting anti-HA. A serologic response to HA Ag was detected in paired sera from patients with type A hepatitis but not from patients with type B or non-A, non-B hepatitis by all three techniques.
Activation of Purified Human Thymus-Derived (T) Cells by Mitogens
Authors: Schmidtke Jon R. , Hatfield Stephen
Published: December 2022

Thymus-derived (T) cells from peripheral blood were purified by rosette formation with neuraminidase-treated sheep red blood cells (SRBC) and centrifugation on Ficoll-Hypaque. T cells recovered from the pellet were freed of SRBC by treatment with Tris-NH4Cl. T cells purified by this method showed a diminished ability to take up 3H-thymidine (3H-TdR) after mitogen stimulation when compared to the mitogenic response of an equal number of autologous peripheral blood mononuclear lymphocytes (PBL). Autologous monocytes restored the capacity of purified T cells to take up 3H-TdR in the presence of phytohemagglutinin (PHA) or Concanavalin A (Con A). The effect was proportional to the number of monocytes added. Similar restorative effects could be obtained with allogeneic or xenogeneic monocytes. These data suggest that the mitogenic stimulation of human PBL and Con A may reflect the participation of more than one cell type: the T cells and monocyte and that the genetic origin of the monocyte is not critical for augmentation of the mitogenic activation of human T cells.
Hapten Concentrations in the Circulation of Animals Actively Immunized with Hapten-Protein Conjugates
Authors: O'Neill Sean P. , Robb Thomas E.
Published: December 2022

Animals immunized with hapten-protein conjugates subsequently circulate high concentrations of hapten bound by antibody. The levels of hapten detected are capable of significantly reducing antibody titer in the sera of immunized animals. In the case of steroid-protein conjugates, the main source of increased plasma steroid concentration is the immunizing conjugate, although a contribution from increased host secretion may also occur.

The results for rabbits immunized with digoxin-BSA indicate that the appearance of circulating digoxin followed the appearance of circulating antibody to digoxin. Appearance of digoxin in circulation appears to coincide with the operation of the immune response and may be related to macrophage activity. Similar conclusions are drawn from results obtained for circulating morphine in the serum of a sheep immunized with morphine-BSA. Injected hapten-protein antigens are probably processed by macrophage to produce low molecular weight haptenic fragments which are maintained in circulation for prolonged periods in the form of antibody-hapten complexes.
The Immune Response to Dextran in BALB/c Mice
Authors: Chen Jenn C. , Leon Myron A.
Published: December 2022

Normal and congenitally athymic mice respond to immunization with dextran, thus indicating that the response is thymus independent. Athymic mice have lower plaque-forming cell (PFC) response than normal mice 3 days after immunization, but similar PFC response as normal mice by 5 days. Seven to 10 days after immunization, athymic mice maintain a high level of PFC response whereas the PFC response declines sharply in normal mice.

Concanavalin A (Con A), added at initiation of in vitro culture, enhances the response of spleen cells from primed normal mice. Treatment with methyl-α-D-mannoside (MAM), a competitive inhibitor of binding of Con A, immediately after addition of Con A, abolishes the enhancement. Treatment with MAM, 3 to 6 hr after Con A addition, partially blocks the enhancement. The enhancing effect is dependent on the dose of Con A and the duration of in vivo priming before in vitro culture.

Con A does not enhance the in vitro PFC response of cells from primed athymic mice. Supernatants from Con A-treated non-primed normal spleen cell cultures enhance the PFC response of “nude” cells, thereby indicating that enhancement by Con A is mediated through a soluble T cell product(s) with no apparent antigen specificity.

Hydroxyurea, an inhibitor of DNA synthesis, blocks the enhancement by Con A of cells primed for 1 to 3 days. Release of hydroxyurea block after 19 hr incubation does not restore the PFC response, thereby indicating the responding anti-dextran PFC precursor cells are engaging actively in DNA synthesis. PFC response of primed cells taken at the peak of anti-dextran response (day 5) shows that they are less susceptible to the hydroxyurea block. Con A enhances the PFC response of cells primed for 5 days even in the absence of DNA synthesis.
Cellular Mechanism of Endotoxin Unresponsiveness in C3H/HeJ Mice
Authors: Glode L. Michael , Scher Irwin , Osborne Barbara , Rosenstreich David L.
Published: December 2022

B cells from C3H/HeJ mice fail to respond to an endotoxin (LPS K235) which is mitogenic for normal mice including the closely related C3H/HeN strain. The cellular basis for this unresponsive state has been investigated. The C3H/HeJ mice have normal numbers of B cells, which are capable of normal responses to other B cell mitogens, such as polyinosinic acid (Poly I). Addition of normal macrophages or spleen cells fails to reconstitute the normal response. Furthermore, neither macrophages nor spleen cells from the C3H/HeJ strain suppress the normal C3H/HeN spleen cells. Finally, spleen cells enriched for B cells by the removal of macrophages or T cells demonstrate the same differences in responsiveness to LPS. These results indicate that LPS unresponsiveness is a defect of the B cell itself and not due to suppressor cells or the absence of helper cells.

When LPS is added to Poly I-stimulated cultures, there is additional enhancement of the response of normal C3H/HeN spleen cells. However, LPS causes a dose-dependent suppression of the Poly I response of C3H/HeJ spleen cells. This suppression is dependent on the time of addition of LPS to the Poly I-stimulated cultures. These data are interpreted as indicating that the binding of LPS to the membrane of C3H/HeJ B cells results in their inactivation or suppression, and that this is the basis of LPS unresponsiveness in this mouse strain.
Regulation of Antibody Response in Vitro
Authors: Kishimoto Tadamitsu , Ishizaka Kimishige
Published: December 2022

The effect of elevation of an intracellular cyclic AMP level on in vitro anti-hapten antibody response was studied, by using mesenteric lymph node cells of rabbits which were primed with dinitrophenylated Ascaris antigen (DNP-Asc) or DNP-ragweed antigen (DNP-Rag). The anti-hapten antibody response was induced by stimulation of the primed B cells by either DNP-heterologous carrier conjugate or anti-immunoglobulin (anti-Ig) for 24 hr (first stage), followed by 6-day culture of the activated cells in the presence of nonspecific enhancing factor (second stage). The stimulation with anti-Ig induced IgG anti-hapten antibody response and enhanced the formation of total IgG. Addition of dibutyryl cyclic AMP or aminophylline with anti-Ig or DNP-heterologous carrier during the first stage enhanced IgG anti-hapten antibody response. The optimal concentration of these reagents for the enhancement was 5 × 10-4 M to 10-3 M. The presence of 5 × 10-6 M prostaglandin E1 during the first stage also enhanced the antibody response. Similarly, the presence of dibutyryl cyclic AMP or aminophylline during the stimulation of DNP-Rag-primed cells with homologous antigen (first stage) enhanced the antibody response. If the same concentration of dibutyryl cyclic AMP or aminophylline was added together with enhancing soluble factor during the second stage after the stimulation of the primed cells with anti-Ig, both the antibody response and the formation of IgG were suppressed. The antibody response of DNP-Rag-primed cells stimulated with homologous antigen was also suppressed if dibutyryl cyclic AMP or aminophylline was added during the subsequent culture (second stage). Evidence was obtained that suppression of antibody response by cyclic AMP during the second stage is probably due to inhibition of the proliferation of B cells. Neither of these drugs suppressed the formation of enhancing soluble factor from the carrier-specific cells stimulated with the homologous carrier. The results obtained in the present experiments suggested that stimulation of hapten-primed B cells with anti-γ chain in the presence of an optimal concentration of dibutyryl cyclic AMP resulted in the formation of a significant amount of IgG anti-DNP antibody without participation of T cells.
Characterization of Cells That Suppress the Cytotoxic Activity of T Lymphocytes
Authors: Clark David A. , Phillips R. A. , Miller R. G.
Published: December 2022

Target cell lysis by sensitized cytolytic T lymphocytes (CTL) may be conveniently quantitated by 51Cr release. By fitting to the formula, P (% specific release) = 100 (1-e-N αt), one obtains α, the relative frequency of CTL in N lymphoid cells. Using a microassay and murine sarcoma target cells, we observed an unexpected decrease in lysis whenever effectors obtained from a graft-vs-host reaction were tested at high concentrations. This inhibition was not observed with CTL generated by an MLC reaction. Inhibition could not be explained by nonspecific mechanical ‘crowding’, reutilization of released isotope, suppression of released from dead target cells, or the particular strain combination and target used. By modifying the formula to allow suppression of CTL by a stochastic cell-cell interaction with a suppressor cell, we found that P = 100 (1-e-N ate- Nγ) adequately fitted the data, where Nγ is proportional to inhibitor content. An 18- to 24-hr incubation at 37°C but not 4°C allowed selective depletion or enrichment of inhibitors; in mixing experiments, both parameters Nαt and Nγ behaved stoichiometrically as independent cellular properties. The inhibitor was resistant to concentrations of anti-T cell (RAMB) serum + complement which killed CTL. A similar inhibitor arose in vivo during an anti-tumour allograft response. The ability to quantitate CTL and inhibitor activities from titration curves provides a technique for studying the identity and mechanism of suppressor cells acting at the effector stage of cell-mediated immunity.
Hereditary C2 Deficiency: Diagnosis and HLA Gene Complex Associations
Authors: Gibson David J. , Glass David , Carpenter C. B. , Schur Peter H.
Published: December 2022

Nine families with genetically controlled C2 deficiency have been described where the propositii and family members are heterozygous C2 deficient. The diagnosis of hereditary C2 heterozygous deficiency was suspected on the basis of analysis for CH50, C2 protein, and C2 function, and then confirmed by family studies. Analysis of HLA antigens in these families supported the close association of C2 deficiency and HLA-A10 and/or B18, particularly the latter. Analysis by MLC studies revealed recombination in one family between the HLA and B and D loci and in another family probable recombination between the D and C2 complement loci. Therefore, the order of the loci on the sixth chromosome is likely to be C2 complement, HLA-D, HLA-B, HLA-A.
Increased Cytolytic T Lymphocyte Activity Induced by 2-Mercaptoethanol in Mixed Leukocyte Cultures: Kinetics and Possible Mechanisms of Action
Authors: Harris John W. , MacDonald H. Robson , Engers Howard D. , Fitch Frank W. , Cerottini Jean-Charles
Published: December 2022

The 20- to 50-fold increase in cytolytic T lymphocyte (CTL) activity caused by the addition of 50 µM 2-mercaptoethanol (2-ME) at the onset of a one-way murine mixed leukocyte culture (MLC) between C57BL/6 and DBA/2 splenic lymphocytes appears to be unrelated to early events in the culture: if 2-ME was present for the first 24 hr of culture only, there was no increase on day 4, but if addition of 2-ME was delayed until the last 24 hr of culture, the CTL activity was almost as high as that of cultures that were exposed to 2-ME for the entire 4-day culture period.

The increase of CTL activity caused by delayed addition of 2-ME (“2-ME rescue”) was used to investigate the mechanism by which the thiol induces differentiation of CTL from precursor cells. 2-ME rescue was mimicked by two other thiols, dithiothreitol and cysteamine phosphate, but at higher concentrations. Because the latter compound has no free sulfhydryl group until it diffuses into cells and is enzymatically dephosphorylated, we conclude that thiols may increase the differentiation of CTL from precursor cells by an intracellular process involving free sulfhydryl groups rather than by interaction with membrane sulfhydryls or destruction of inhibitor cells or their products.

Cell separation experiments indicated that 2-ME rescue was independent of the presence of B lymphocytes and of adherent cells (macrophages) and was restricted to a subpopulation of T lymphocytes that developed into large lymphoid precursor cells during the first 3 days in culture even without 2-ME. The development of this subpopulation required DNA synthesis between 24 and 72 hr after the onset of MLC. When 2-ME was added to day -3 MLC, CTL activity increased slightly as early as 4 hr later, but the major increase occurred during the second half of the 24 hr “rescue” period. Because this increase was inhibited by cytosine arabinoside (ARA-C), it seems likely that DNA synthesis is associated with and may be required for the differentiation of large precursor lymphoid cells into CTL after the addition of 2-ME.
A Phosphorycholine Idiotype Related to TEPC 15 in Mice Infected with Ascaris Suum
Authors: Brown Alan R. , Crandall Catherine A.
Published: December 2022

A serum component, binding antigens having phosphorylcholine (PC) determinants were induced in several strains of mice by infection with Ascaris suum. This component was isolated and demonstrated to be an IgM (κ) anti-PC antibody having idiotypic determinants in common with the IgA PC-binding myeloma protein TEPC 15. Rabbit anti-idiotypic antisera prepared with this component had idiotypic specificity for TEPC 15 and cross-idiotypic recognition of MOPC 167 and McPC 603, all IgA PC-binding myeloma proteins. The antisera also recognized determinants not present on TEPC 15.

IgM and idiotype levels were quantitated by radial immunodiffusion and PC-specific antibody measured by hemagglutination (HA) with sheep erythrocytes coated with pneumococcal-C-polysaccharide. Mean IgM levels ranged from 2.5 to 8.7 mg/ml, idiotype from 0.54 to 5.3 mg/ml; and HA titers from 1:512 to 1:130,000 in different mouse strains. The high PC-specific antibody response was duplicated by immunization with dead ascaris larvae or by infection with two other nematode species.
Studies on Tolerance Induction in Vitro
Authors: Schaefer Annette E. , Fishman Marvin , Adler Louise T.
Published: December 2022

The induction of specific immunologic tolerance and its transfer has been studied in the in vitro response of unprimed rabbit spleen cells to solubilized phage antigens. Evidence presented shows that only viable cells transfer tolerance and that the responsible cells appear devoid of membrane-bound antigen but are sensitive to anti-thymocyte serum. It appears, therefore, that although the in vitro response to solubilized S-T2 has no demonstrable requirement for T “helper” cells, it is subject to the action of T “suppressor” cells.
Temporary Inhibition of Antibody-Dependent, Cell-Mediated Cytotoxicity by Pretreatment of Human Attacking Cells with Ammonium Chloride
Authors: Yust Israel , Smith Richard W. , Wunderlich John R. , Mann Dean L.
Published: December 2022

Treatment of lymphoid cell preparations with ammonium chloride buffers to remove contaminating erythroid cells markedly reduced the ability of these cells to function as attacking cells in antibody dependent cell mediated cytotoxicity tests (ADCC). The lymphoid cells, however, retained their ability to adhere to antibody-coated target cells Recovery of function 20 hr after exposure to ammonium chloride suggests a metabolic rather than receptor effect of these salts on the ADCC function of lymphocytes.
The Induction of Hapten-Specific Immunologic Tolerance and Immunity in B Lymphocytes
Authors: Fidler John M.
Published: December 2022

An improved adoptive transfer procedure has been developed for the thymus-independent conjugate TNP-LPS. This technique involving delayed immunization results in increased sensitivity and makes possible studies of various cell types, the response of which is normally very low or which are difficult to manipulate experimentally in situ.

The standard adoptive immune response of adult spleen cells to TNP-LPS was low in comparison to the primary AFC response of intact mice. Adult bone marrow cells gave a 30-fold lower and neonatal spleen cells an 8-fold lower adoptive response than an equivalent number of adult spleen cells. If the administration of antigen was delayed past the normal time of immunization (1 hr after cell transfer), the resulting AFC response of adult spleen, adult bone marrow, and neonatal spleen cells was enhanced. The peak response occurred with a delay interval of 4 days, and the magnitude decreased with greater delay. Part of the reason for the decline was an acceleration in the attainment of a peak response in mice given antigen past the 4-day optimum. The enhancement of the adult splenic response was not restricted to the antigen TNP-LPS, and was a transferrable phenomenon which did not require antigen to persist past the 4-day optimum delay period. The ineffectiveness of host pre-irradiation indicated that host recovery alone was not involved. Although the results are more compatible with B cell proliferation causing the enhancement, a combination of effects may be involved.
Two Immunochemically and Physicochemically Distinct Secretory Components from Rat Exocrine Secretions
Authors: Cambier John C. , Butler J. E.
Published: December 2022

Two proteins were isolated from rat milk by using ion-exchange and gel filtration chromatography and shown to be secretory components by the criteria of: a) occurrence as free proteins in exocrine secretions, as well as bound to exocrine IgA while absent from serum and serum IgA, b) immunohistochemical localization in the epithelial cells of the gut, c) dissociability from and in vitro binding to 11 S SIgA, d) cross-reactivity with antisera to mouse and bovine secretory components, and e) significant peptide homology with bovine secretory component.

The secretory components were immunochemically and physicochemically distinct as shown by: a) the lack of cross-reactivity between their specific antisera and differential reactivity with anti-bovine FSC and anti-mouse SC, b) different electrophoretic mobilities, c) DEAE elution profiles, d) carbohydrate contents, e) molecular weights, and f) apparent polypeptide chain constitution. Another interesting difference in the two is that only one of the components dissociates spontaneously from SIgA. Possible functions of two secretory components are discussed.
Induction of Immunologic Tolerance in Nursing Neonates by Absorption of Tolerogen from Colostrum
Authors: Halsey John F. , Benjamin D. C.
Published: December 2022

Deaggregated human γ-globulin (DHGG) injected into female mice within 24 hr after delivery of a litter enters the colostrum and is absorbed intact through the intestine by nursing neonates. This absorbed HGG was present in the neonatal circulation at concentrations of 0.3 to 0.6 mg/ml of serum under the experimental conditions used. This absorption of HGG by the nursing neonate resulted in a complete, specific, tolerant state to HGG. This tolerant state was stable upon adoptive cell transfer and could not be abrogated by transfer of normal syngeneic spleen cells.
Local Immunization in the Mammary Glands of the Rabbit
Authors: Hurlimann Jean , Lichaa Mireille
Published: December 2022

The effects of a local immunization with bovine serum albumin in the left mammary glands of the rabbit have been compared with those of an intramuscular (i.m.) immunization. The ability of several tissues to synthesize antibodies was studied in vitro with or without in vitro antigenic stimulation, and the antibody class in the plasmocytes of these tissues was determined by immunofluorescence. After i.m. immunization, the serum antibodies synthesized by the spleen and the lymph nodes pass into the milk by transudation; there is no antibody synthesis in the mammary glands. After immunization in the left mammary glands, there is antibody synthesis in the left mammary glands and also in the spleen and lymph nodes. In the mammary glands the antibodies belong essentially to the IgG class and are synthesized by the plasmocytes that infiltrate the granulation tissue around the injection sites. The fragments of the immunized glands are capable of an in vitro secondary response analogous to that made by the spleen and lymph nodes. Thus the local immunization of the mammary glands does not stimulate the secretory immunologic system but elicits an antibody synthesis in the granuloma as in any other tissue. This local immunization also provokes a traffic of sensitized cells to the same lymphoid tissues as an i.m. immunization does.
The Suppressive Effect of Immunization on the Proliferative Responses of Rat T Cells in Vitro
Authors: Bash Jerry A. , Singer Andrew M. , Waksman Byron H.
Published: December 2022

Rats given large i.v. doses of ovalbumin or sheep erythrocytes manifest suppressed spleen cell responses (3H-thymidine incorporation) to PHA within hours. Removal of glass wool-adherent cells totally restores responsiveness to that of normal nonadherent spleen cell cultures. Carrageenan, selectively toxic for macrophages, partially restores responses of antigen-suppressed spleen cells in culture, suggesting a supportive role for macrophages in the suppression phenomenon. Treatment of donors with low doses of cyclophosphamide (20 to 50 mg/kg) at the time of antigen injection abrogates the ability of their spleen cells to suppress the responses of normal cells to PHA. The low dose of cyclophosphamide required indicates a target other than the B cell or macrophage and suggests the possibility that cyclophosphamide eliminates the suppressor T cell component of the macrophage-T cell complex.
Antigen-Specific and Nonspecific Mediators of T Cell/B Cell Cooperation
Authors: Harwell Lee , Kappler John W. , Marrack Philippa
Published: December 2022

T cell-containing lymphoid populations produce a nonantigen-specific mediator(s) (NSM) which can replace T cell helper function in vitro in the response of B cells to sheep red blood cells (SRBC), but not to the hapten-protein conjugate, trinitrophenyl-keyhole limpet hemocyanin, (TNP-KLH). NSM produced under three conditions: 1) stimulation of KLH-primed cells with KLH; 2) allogeneic stimulation of normal spleen cells; and 3) stimulation of normal spleen cells with Con A (but not PHA) are indistinguishable on the basis of their biologic activity and m.w., estimated as 30 to 40,000 daltons by G-200 chromatography. Production of NSM is dependent on the presence of T cells. The action of NSM on B cells responding to SRBC in the presence of 2-mercaptoethanol is unaffected by severe macrophage depletion. Extensive absorption of NSM with SRBC failed to remove its activity, confirming its nonantigen-specific nature.