bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of Activated Macrophages in Specific and Nonspecific Cytostasis of Tumor Cells
Authors: Krahenbuhl James L. , Remington Jack S.
Published: December 2022

Peritoneal macrophages but not peritoneal lymphocytes from mice chronically infected with the intracellular protozoan parasites Toxoplasma gondii and Besnoitia jellisoni had a marked cytostatic effect on a variety of tumor target cells. Cytostasis was quantitatively measured as an inhibition in uptake of tritium labeled thymidine (3H-TdR) by L cells, EMT-6 adenosarcoma, and lymphomas syngeneic for DBA/2 and C57BL mice within 6 hr after culture in vitro with protozoal-activated macrophages. Slight or no cytostasis was effected by normal macrophages from unstimulated mice or mice injected i.p. with starch or thioglycollate. Whereas the cytostatic capabilities of protozoal activated macrophages were directed nonspecifically against several types of tumor target cells regardless of histocompatability, specific cytostasis was demonstrated by macrophages from inbred mice immunized with allogeneic mouse lymphoma cells.
Mouse Strain-Specific Idiotypy and Interstrain Idiotypic Cross-Reactions
Authors: Briles David E. , Krause Richard M.
Published: December 2022

Genetic studies with the crosses and backcrosses of SWR/J and A/J mice indicate that the idiotypic determinants of Group A streptococcal antibodies segregate in a Mendelian fashion. The idiotypic reactions were inhibited with phenyl-β-N-acetyl-glucosaminide which contains immunodominant determinants of the Group A carbohydrate.

Some of the absorbed idiotypic antisera to SWR/J and A/J Group A antibodies failed to detect strain-specific idiotypic determinants. Such anti-idiotypic sera exhibited interstrain idiotypic cross-reactions with the Group A antibodies from a number of different mouse strains. These idiotypic cross-reactions were also inhibited by phenyl-β-N-acetyl-glucosaminide, a finding which suggests that these interstrain idiotypic cross-reactions are dependent at least in part on site-associated determinants.
Reaginic Antibody Formation in the Mouse
Authors: Okudaira Kirokazu , Ishizaka Kimishige
Published: December 2022

An adoptive transfer system for an IgE anti-hapten antibody response was established in DBA/1 mice with dinitrophenylated ovalbumin (DNP-OA) and keyhole lympet hemocyanin (DNP-KLH) as antigens. Maximal anti-DNP IgE antibody response was obtained by transfer of DNP-primed cells and carrier-specific cells for the secondary carrier to irradiated syngeneic mice, followed by challenge with DNP-secondary carrier conjugate in saline, a procedure which does not induce a primary IgE antibody response. Recipients of hapten-primed cells or carrier-specific cells alone showed meager or no IgE antibody response. Effect of aluminum hydroxide gel (alum) with antigen for carrier immunization and hapten priming was studied by the adoptive transfer technique. It was found that immunization of donors of carrier-specific cells with OA in saline, which did not induce either IgG or IgE antibody response, resulted in a high level of helper function for both IgE and IgG anti-hapten antibody responses in the adoptive response of DNP-KLH-primed cells to DNP-OA. Similarly, addition of a low dose alum with KLH was sufficient for carrier priming for helper function with respect to the anti-DNP IgE antibody response. It was shown, however, that alum had a significant effect on hapten priming. Recipients of the spleen cells from donors primed with DNP-OA or DNP-KLH plus a high dose (1 mg) alum gave much higher anti-hapten IgE antibody response than the recipients of DNP-primed cells obtained by priming with the same protein in saline. Animals primed with DNP-KLH with a low (50 µg) dose alum gave IgG but no IgE secondary response. In spite of the presence of both carrier-specific helper cells and hapten-primed precursors for IgE antibody response, these animals gave rise to a secondary IgG antibody response but failed to show IgE antibody response upon a booster injection with DNP-KLH in saline. If their spleen cells were transferred to irradiated animals, the recipients showed high IgE and low IgG antibody responses upon injection of DNP-KLH in saline. These results suggest that secondary IgE antibody response is enhanced in irradiated recipients.
A New One-Step Method for the Functional Assay of the Fourth Component (C4) of Human and Guinea Pig Complement
Authors: Gaither Thelma A. , Alling David W. , Frank Michael M.
Published: December 2022

A simple one-step method for the molecular titration of functional C4 has been developed. This method utilizes serum from the C4D strain of guinea pigs as a reagent supplying all components except C4 in relative excess. Guinea pig C4 titers obtained with this method are slightly higher than titers obtained with classical functional method. In assay of human C4, titers obtained are more than 4-fold higher than those obtained with the classical functional method.

The correlation between the classical and C4D methods of C4 titration is excellent and the C4D method has less inherent variability as shown by an analysis of variance. Some of the factors which determine the end point in C4 titration by this method have been examined in detail as has been the usefulness of the method for sera of various species. The method has been used in a longitudinal study of C4 levels in nine normal subjects. In this group the range of normal titers was wide. Most, but not all, individuals showed remarkable stability in their C4 titers over long periods of time.
In Vitro Antibody Formation by Human Tonsil Lymphocytes
Authors: Watanabe Takeshi , Yoshizaki Kazuyuki , Yagura Takayasu , Yamamura Yuichi
Published: December 2022

In vitro antibody formation by human lymphocytes was observed against hapten-protein conjugates by using human palatine tonsil cells in the presence of an optimum concentration of 2-mercaptoethanol. Relatively pure populations of B and T lymphocytes were prepared by the method of Ficoll-Hypaque gradient centrifugation to investigate cellular mechanism of antibody formation by human lymphocytes. Maximum response to hapten-protein conjugates was observed in vitro when T and B cells were mixed at an optimal ratio. Little response occurred when each cell alone was cultured with antigen. Depletion of glass-adherent cells also resulted in diminution of the anti-hapten response. These data indicated that three kinds of cells—T cells, B cells, and glass-adherent cells—are essentially necessary for in vitro primary antibody formation by human tonsil cells.
Lymphocyte Plasma Membranes
Authors: Zimmerman Barry
Published: December 2022

In the present study it was shown that heterologous anti-human lymphocytic sera (ALS) specifically bound isotopically labeled surface antigens. The antigen-binding capacity of ALS was quantitative and different antisera had differing potencies and specificities. The antigens bound by ALS were species-specific when cross-reactivity was tested between phylogenetically distant species. There was, however, partial sharing of the lymphocyte antigens with human liver, kidney, and brain, although not with red cells and serum. Antigenic differences were demonstrated between human thymus lymphocytes (THY) and cultured human lymphoblasts (CHL) with ALS raised against thymus [ALS(THY)] or lymphoblasts [ALS(CHL)]. Differences between the antigens bound from labeled THY and labeled CHL were also noted on polyacrylamide-SDS gel electrophoresis of the solubilized precipitates. When lymphocytes from peripheral blood (PBL) and tonsil (TON) were compared to THY and CHL with ALS (THY) and ALS (CHL) the PBL resembled THY whereas TON gave results intermediate to THY and CHL.
Immunologic Reactions to Haptens on Autologous Carriers
Authors: Walters Curla S. , Claman Henry N.
Published: December 2022

Specific tolerance to the hapten NIP was induced in mice with a highly substituted conjugate the NIP-MGG (NIP-mouse γ-globulin). A correlation was made between tolerance at the serum level (as measured by the absence of NIP antibody) and tolerance at the cellular level as measured by absence of antigen-specific DNA synthesis. Different schedules were used for tolerance induction. Hapten-specific tolerance could be induced if soluble NIP-MGG was injected from 8 days before to 2 days after immunogen. At these times, tolerance was seen by both depression of serum anti-NIP antibody and suppression of antigen-driven DNA synthesis in vitro.

Both partial cross-stimulation and partial cross-tolerance were seen. In cross-stimulation, cells from mice immunized with NIP-MSA responded in vitro to NIP-MGG-this response was greatly reduced if the mice were also tolerized in vivo with soluble NIP-MGG. In cross-tolerance, cells from mice immunized with NIP-MSA and tolerized with NIP-MGG showed a reduced response to NIP-MSA in vitro.

We interpret these reults to mean that NIP-specific cells were generated in vivo by immunization, that these cells also responded to hapten in vitro, and that they could also be tolerized in vivo. In addition, however, there are cells specific for new antigenic determinants (NAD) formed by coupling NIP to carriers. These NAD-specific cells can also be generated in vivo and stimulated in vitro but are not susceptible to cross-stimulation or cross-tolerance by NIP on other carriers.
Radioresistance of Some Biologic Properties of Mouse Mast Cells
Authors: Kojima Somei , Ovary Zoltan
Published: December 2022

Mast cells taken 11 or 25 days after a sublethal irradiation (540 R) formed rosettes with antibody-coated sheep erythrocytes in the same number as in control non-irradiated mice. The threshold of passive cutaneous anaphylaxis reactions with mouse IgG1 or IgE antibody 2 to 7 days after irradiation was the same in sublethally (540 R), or lethally (780 R), irradiated mice as it was in controls.
The Effect of Estrogen on the Incorporation of 3H-Thymidine by PHA-Stimulated Human Peripheral Blood Lymphocytes
Authors: Ablin Richard J. , Bruns Gailon R. , Guinan Patrick , Bush Irving M.
Published: December 2022

The incorporation of 3H-thymidine by PHA-stimulated human peripheral blood lymphocytes from healthy adult males was significantly (p 0.002) suppressed in the presence of diethylstilbestrol diphosphate. The observed suppression of blastogenesis in the presence of exogenous estrogen suggests that the palliative effects of estrogenic therapy in the clinical treatment of patients with hormonally-responsive tumors may be countered by its adverse effect of the host's cell-mediated immunologic responsiveness.
Basophilic Leukocytes in Delayed Hypersensitivity Reactions
Authors: Katz Stephen I. , Heather Christopher J. , Parker Darien , Turk John L.
Published: December 2022

In guinea pigs, basophilic leukocytes constitute a high proportion of the cells infiltrating allergic contact, Jones-Mote and tuberculin-like reactions, when elicited 1 week after sensitization with 2,4 dinitrofluorobenzene, ovalbumin (OA) in Freund's incomplete adjuvant (FIA) and OA in Freund's complete adjuvant (FCA), respectively. When animals are pretreated with one dose (300 mg/kg) of cyclophosphamide, 3 days before sensitization, allergic contact and Jones-Mote reactions, elicited 7 days after sensitization, are increased in intensity and induration and prolonged, whereas the tuberculin-like reactions are somewhat reduced in intensity. The percentages of basophils infiltrating all these altered reactions are markedly reduced. Thus, delayed-in-onset skin reactions may occur in guinea pigs immunized with proteins in FIA or FCA or in contact-sensitized animals in the presence or absence of basophils.
Immunologic Functions of Isolated Human Lymphocyte Subpopulations
Authors: Chess Leonard , MacDermott Richard P. , Schlossman Stuart F.
Published: December 2022

A Sephadex G-200 immunoabsorbent column bound with pure rabbit antihuman Fab was used to fractionate human peripheral lymphocytes. Two distinct populations of cells were quantitatively recovered. The cells passing directly through the column without binding were T cells, as judged by both immunofluorescent and E rosetting properties. On the other hand, the retained cells which could be specifically and quantitatively eluted with human Ig were τ;97% B cells, as judged by the fluorescence and rosetting criteria. Both T and B cells incorporated 3H-thymidine significantly in response to phytohemagglutinin, concanavalin A, and pokeweed mitogen, while neither population proliferated in response to endotoxin or antihuman Fab. The blast cells induced by mitogen in T or B cell cultures retained their cell surface characteristics. T cell blasts were devoid of surface Ig while τ;90% B cell blasts had Ig on their cell surface.
Antibody-Induced Redistribution of Measles Virus Antigens on the Cell Surface
Authors: Joseph Barry S. , Oldstone Michael B. A.
Published: December 2022

Antibody specific for measles virus can redistribute (cap) measles virus antigens on infected cells in culture. Capping of measles virus antigens at the cell surface requires active cell metabolism and multivalent antibody. Capping was inhibited with oligomycin D, vinblastine sulfate, colchicine, and cytochalasin B, suggesting that ATP production and functioning microfilaments and microtubles are involved in the capping process.
Humoral and Cell-Mediated Immune Responses to Two Hepatitis B Virus Antigens in Guinea Pigs
Authors: Gerety Robert J. , Hoofnagle Jay H. , Barker Lewellys F.
Published: December 2022

The ability of two purified, particulate hepatitis B virus antigens to induce antibodies and delayed cutaneous hypersensitivity responses was studied in Hartley guinea pigs. Hepatitis B surface antigen (HBsAg) was purified by several methods from the plasmas of human, chronic HBsAg carriers, while hepatitis B core antigen (HBcAg) was purified from chimpanzee liver by ultracentrifugation. Guinea pigs immunized with the HBsAg preparations developed anti-HBs in the absence of anti-HBc but delayed cutaneous hypersensitivity reactions to HBsAg were seen clearly only in animals immunized with high doses of the most highly purified HBsAg preparation. Guinea pigs immunized with the HBcAg preparation developed anti-HBc in the absence of anti-HBs, and showed delayed cutaneous hypersensitivity reactions to HBcAg in the absence of reactions to a chimpanzee liver-derived control antigen. No skin reactivity was seen to HBcAg in guinea pigs immunized with HBsAg. Similarly, guinea pigs immunized with HBcAg showed no skin reactivity to HBsAg. It was concluded that both HBsAg and HBcAg are capable of inducing distinct humoral and cell-mediated immune responses in Hartley guinea pigs.
Suppressive Effect of Interferon on the Humoral Immune Response to Sheep Red Blood Cells in Mice
Authors: Brodeur Bernard R. , Merigan Thomas C.
Published: December 2022

Swiss-Webster mice injected intravenously with interferon preparations 2 days before primary or secondary immunization with sheep erythrocytes showed a significant suppression of their hemolysin and agglutinin responses as measured in microtiter assays. Interferon treatment in vivo also inhibited DNA synthesis induced in mouse lymphocytes studied in vitro by concanavalin A, phytohemagglutinin, and lipopolysaccharide. In addition, it suppressed the in vitro primary and secondary response to sheep erythrocytes. It is possible that this inhibition of the immune response by interferon is significant in the immunosuppression observed during viral infection or after administration of nonspecific mitogens, or perhaps even in the control of the normal immune response.
Development of Humoral Immunity in Lethally Irradiated Mice Reconstituted with Fetal Liver
Authors: Sherwin William K. , Rowlands David T.
Published: December 2022

A model system of ontogeny was utilized to investigate the development of humoral immunity. Lethally irradiated adult C3H mice were reconstituted with syngeneic fetal liver. These mice were immunized at various times after reconstitution with a series of eight antigens: the bacteriophages F2 T4 and øX-174; the hapten carrier complexes 2,4 dinitrophenyl-bovine serum albumin and fluorescein bovine serum albumin; and the small proteins, hen egg lysozyme, sperm whale myoglobin, and bovine pancreatic ribonuclease. Subsequent antibody production to the bacteriophage was assayed using a bacteriophage neutralization technique. To detect antibody directed against the haptens or small proteins, the appropriate hapten or protein was first coupled to T4, and then a modified bacteriophage neutralization technique was employed. Individual mice responded to the various antigens in a sequential pattern which was basically the same for all mice within the strain. This hierarchy of responsiveness could not be explained on the basis of different immunogenicities of the antigens nor on different sensitivities of the antibody assays. Instead, we propose that the hierarchy results from the ordered maturation of lymphopoietic elements in the fetal liver inoculum.
Complement-Induced Histamine Release from Human Basophils
Authors: Siraganian Reuben P. , Hook William A.
Published: December 2022

The activation of human serum complement by incubation with zymosan generates C5a which releases histamine from autologous basophils. The characteristics of the C5a-induced histamine release were investigated. It is similar to IgE-mediated reactions in requiring Ca++ and in being inhibited by EDTA. However, it has marked differences from IgE-mediated reactions. C5a, at all concentrations, released histamine completely in less than 2 min. The C5a reaction has a narrower pH optimum than antigen-induced release and occurs well at 17° to 37°C but not at 0°C. The optimal reaction temperature is 25° to 30°C. Unlike the antigen-induced release, no two-stage activation with C5a for the release of histamine could be demonstrated. There was additive release between C5a- and IgE-mediated reactions. Leukocytes could be desensitized to the C5a-mediated reaction by 1) incubating the cells at 37°C for 45 min, 2) pretreating the leukocytes with activated serum in the presence of EDTA, and 3) adding the activated serum to the leukocytes at 0°C before transferring to the optimal reaction temperatures. Cells desensitized to the complement-induced release have normal reactions to IgE-mediated histamine release. In parallel experiments, cells from allergic donors desensitized for IgE-mediated reactions by incubation with antigen under sub-optimal conditions release histamine normally upon the addition of C5a. The results indicate that histamine release by C5a involves a mechanism of basophil activation that is different from the pathway involved in the IgE-induced reaction.
Tolerance and Contact Sensitivity to DNFB in Mice
Authors: Claman Henry N.
Published: December 2022

Immunologic unresponsiveness (tolerance) was induced in a mouse model of contact sensitization to DNFB. The ability to induce tolerance varied with the chemical reactivity of the tolerogen; DNFB was highly tolerogenic, DNBSO3 was moderately tolerogenic, and DNP-lysine was not tolerogenic. Although DNFB is considered a highly reactive compound, tracer studies of injected DNFB* showed that it was rapidly excreted. Further studies were therefore done with DNFB attached to mouse erythrocytes. Tolerance to DNFB-RBC was highly specific in vivo; mice tolerant to DNFB showed normal reactivity to TNCB (picryl chloride). Cells of mice tolerant to DNFB-RBC were also unresponsive to DNBSO3 in vitro. Tolerance to DNFB, DNBSO3, and DNFB-RBC all required time to develop, suggesting that an active process was involved.
Tumor-Associated Blocking Factors: Isolation from Sera of Tumor-Bearing Mice
Authors: Tamerius J. , Nepom J. , Hellström I. , Hellström K. E.
Published: December 2022

An isolation and partial purification of tumor-associated blocking factors from the sera of tumor-bearing mice is described. Columns for affinity chromatography were prepared by coupling syngeneic tumor-immune antibodies to Sepharose 4B. Passage of serum through such immunoadsorbent columns removed all blocking activity from tumorbearers' sera; subsequent elution of the absorbent with 3 M NaSCN recovered the activity. The blocking material was further purified on Sephadex G-200. The data provide evidence for the presence of antigen in tumor-associated blocking factors and are compatible with the hypothesis that blocking factors often consist of antigen and antibodies in the form of immune complexes.
Influence of Protein Restriction on Immune Functions in NZB Mice
Authors: Fernandes Gabriel , Yunis Edmond J. , Good Robert A.
Published: December 2022

The influence of a low protein (6%) diet on the immunologic function of NZB mice was investigated. The low protein intake was associated with decreased weight gain in both male and female NZB mice. The mice fed the low protein diet did not develop splenomegaly, which generally occurs by 7 to 10 months of age in NZB mice fed a normal amount of protein. Further, 7- to 10-month-old NZB mice fed the low protein (6%) diet, maintained: 1) more vigorous antibody production to sheep red blood cells; 2) greater capacity to produce graft-vs-host reactions, and 3) more vigorous cell-mediated “killer” cell immunity after immunization against DBA/2 mastocytoma cells than did NZB mice on a normal (22%) protein diet. The decrease of PHA and Con A response which normally occurs with aging in NZB mice was abrogated to some degree by protein restriction. However, response to LPS, which also declines with age in NZB mice, did not appear to be influenced by diet. The low protein diet inhibited the rise of IgG1 levels, prevented the increase of IgM levels and the fall of IgA levels. The low protein diet, although delaying the onset of hemolytic anemia, did not prevent the development of this autoimmune disease and did not significantly prolong life span.

These findings tend to dissociate the progressive deficit of T cell immunity in NZB mice from the genetically determined propensity to develop autoimmune hemolytic anemia. The findings further indicate that it is not yet possible to generalize concerning the relationship of nutrition to deficiencies in immunologic function.
Regulation of the Immune Response to Tumor Antigens
Authors: Fujimoto Shigeyoshi , Greene Mark I. , Sehon Alec H.
Published: December 2022

Immunosuppressor (IS) cells were found to be generated in tumor-bearing animals (TBA) within 24 hr after inoculation of tumor cells of the methylcholanthrene-induced Sarcoma 1509a and appeared to persist in the hosts as long as the tumor was progressing. However, IS cells disappeared with 5 days after extirpation of the tumor.

Increasing doses of thymus cells of TBA increased the degree of suppression of tumor rejection in immune syngeneic animals. Ten million thymus cells of TBA were capable of suppressing significantly the tumor rejection. The IS cells were detected in the thymuses, spleens, and draining lymph nodes, as well as in bone marrow of TBA, but could not be detected in the peripheral circulating blood. Since immunosuppressive activity of bone marrow cells from TBA was entirely abolished by the in vitro treatment of the cells with anti-θ serum and complement, the observed immunosuppression appears to be mediated by the T cell population.

Hydrocortisone treatment of TBA did not abolish the suppressive activity of the thymus or spleen cells of the treated TBA, thus suggesting that IS cells were cortisoneresistant.

The suppressive activity was found to be associated with cells of lowest density on centrifugation through a discontinuous density gradient of Ficoll. The results of this study, taken together with those presented in the preceding article, suggest that IS cells belong to a T cell subpopulation (immunosuppressor T cells). The relationships between immunosuppressor T cells and other T cell subpopulations, i.e., cytotoxic (effector) or helper (amplifier) T cells, remain to be elucidated in further experiments.
Production of Goat, Rat, and Guinea Pig Antisera to Murine IgE
Authors: Lang G. M. , Lee W. Y. , Carter B. G. , Sehon A. H.
Published: December 2022

The study of immunoglobulins (Ig)3 has been facilitated by the production of specific anti-Ig sera (1–4). For each of the six of the known classes and subclasses of mouse Ig, there exists a myeloma synthesizing the corresponding paraprotein, which has been used for the preparation of specific antiserum. Myeloma proteins have also been used to study the IgE class of antibodies in man (5) and in the rat (6), but to date no mouse IgE (IgEm) myeloma has been discovered. Nevertheless, one report has been published in which it was claimed that rat anti-mouse IgEm was produced (7) by immunization of Lewis rats with syngeneic peritoneal cells coated with IgEm; the mouse reagin has been adsorbed onto these cells by incubating them with mouse reaginic serum (MRS). However, in spite of extensive efforts, this procedure could not be reproduced in this laboratory.
Augmentation of Cell-Mediated Cytotoxicity Against Syngeneic Gross Virus-Induced Lymphoma in Rats by Phytohemagglutinin and Endotoxin
Authors: Glaser Moshe , Djeu Julie Y. , Kirchner Holger , Herberman Ronald B.
Published: December 2022

Normal W/Fu rat spleen cells cultured for 1 day with mitogenic concentrations of phytohemagglutinin (PHA) or with a wide range of concentrations of endotoxin (LPS) exerted cytotoxic activity against the syngeneic Gross virus-induced lymphoma, (C58NT)D, and also against other tumor cells in a 4-hr 51Cr release assay. This activity did not involve release of detectable lymphotoxins, nor did it require the presence of additional mitogen in the media during the cytotoxicity assay. The cytotoxic activity induced by PHA was dependent on the presence of T cells while the activity induced by LPS was not dependent on T cells and appeared to require the presence of macrophages. Both PHA and LPS also augmented the cytotoxic activity of spleen cells immune against (C58NT)D tumor cells. With submitogenic concentrations of PHA, only specific cytotoxicity against (C58NT)D cells was augmented. In contrast, LPS and mitogenic concentrations of PHA caused nonspecific augmentation. Mitogenic concentrations of PHA and LPS also augmented the antibody-dependent cytotoxicity of normal W/Fu rat spleen cells against (C58NT)D cells in the presence of syngeneic antiserum.
Anti-Immunoglobulin Stimulation of Murine Lymphocytes
Authors: Weiner Howard L. , Moorhead John W. , Claman Henry N.
Published: December 2022

The in vitro proliferative response of normal mouse spleen cells to anti-immunoglobulin (Anti-Ig) reagents was found to be an age-associated phenomenon. The response usually appears in mice when they reach 5 to 7 months of age and is rarely seen in younger animals. Anti-Ig induced proliferation was observed by using two different antisera—one polyvalent, prepared against mouse antibody-antigen complexes and one prepared against mouse IgM myeloma. Both antisera were shown to be specific for B cells by cytotoxicity and immunofluorescent staining. Glassbead separation of spleen cells showed that the anti-Ig-induced proliferation was a B cell response.
Studies on the Antigenic Determinants of the Thy-1.2 Alloantigen as Expressed by the Murine Lymphoblastoid Line S-49.1 TB·2·3
Authors: Johnson Brian J. , Kucich Umberto N. , Maurelli Anthony T.
Published: December 2022

Sialic acid has been shown to be part of the antigenic determinant of the Thy-1.2 alloantigen as expressed by the murine cell line S-49.1 TB·2·3 (S-49). This conclusion is based on the loss of cytotoxic inhibitory activity by the action of neuraminidase on the Thy-1.2 alloantigen. Also, sialic acid has the ability to inhibit the cytotoxic assay of AKR anti-C3H Thy-1.2 serum for S-49 cells. Further evidence for the protein nature of the Thy-1.2 alloantigen is apparent by trypsin digestion. Possibly the Thy-1.2 alloantigen as expressed on S-49 cells is a glycoprotein.
Suppression of Antibody Responses in Allogeneic Mice by Products of Lymphoid Tissue
Authors: Yonkosky Donna , Buffett Rita F. , Bennett Michael
Published: December 2022

Mice were irradiated, infused with thymocytes and immunized with a variety of antigens, i.e., sheep or horse red blood cells (SRBC or HRBC), diphtheria toxoid (DT) or bovine γ-globulin (BGG). The spleen cells (T.Spleen cells) were harvested 5 days later and cellfree extracts were prepared. The extracts contained an allogeneic suppressive factor (ASF) that was capable of inhibiting IgM antibody responses of allogeneic or semi-allogeneic unirradiated mice. ASF had to be injected within 24 hr of immunization to be effective and a single injection delayed, rather than abolished, the antibody response at the cellular level. However, daily injections of ASF resulted in persistent suppression of antibody response. ASF activity was antigen nonspecific, i.e., the antigen used to stimulate ASF production did not have to be the same as the antigen used to test for ASF activity. C3H T.Spleen extracts were even immunosuppressive when prepared by exposure to C3BF1 alloantigens only; such extracts suppressed antibody responses of C3BF1 and DBA/2 mice.

C3H ASF was removed from extracts after incubation with C3BF1 spleen cells but not after incubation with C3H spleen cells. C3BF1 spleen cells which had been preincubated with C3H ASF were unable to generate antibody-forming cells upon transfer to irradiated C3BF1 host mice. This suggests that the ASF molecule may be or include receptors for alloantigens.

The immunogenetic requirements for ASF activity were evaluated by injecting extracts from C3H, C57BL, C3BF1 and BALB/c T.Spleen cells into C3H, CBA, C57BL, BALB/c, DBA/2, A or C3H.A recipient mice. All extracts tested had ASF activity. However, all allogeneic recipients were not suppressed by the extract material. The suppressive activity of ASF seemed to require two (or more) antigenic differences between donors and recipients of extract material, an H-2K or I antigen difference and a second antigen difference, possibly Ig-1. In the limited numbers of strain combinations tested, T.Spleen extracts suppressed IgM antibody response only if exposed to H-2 and Ig-1 antigens, e.g., BALB/c (H-2d, Ig-1a) ASF suppressed A (H-2a, Ig-1e) but not C3H.A (H-2a, Ig-1a) or DBA/2 (H-2d, Ig-1c). Separate ASF molecules may react with separate antigens on the cell surface, i.e., with H-2 and γG2a. Alternatively, one ASF molecule may react with two structurally associated antigens. If the latter is correct, it is conceivable that the β2-microglobulin which is non-covalently linked to the major component of H-2 molecules expresses allotypic antigens coded for by Ig-1 and β2-microglobulin is one of the antigens recognized by ASF.
A Receptor for Activated C3 in the Normal and Abnormal Human Renal Glomerular Capillary Wall
Authors: Gelfand M. C. , Shin M. L. , Frank M. M. , Nagle R. , Green I.
Published: December 2022

Most immunologically mediated renal disease results from the deposition of antigen-antibody-complement complexes within the glomerular capillary wall, yet the cause of this selective deposition is unknown. It has been shown by Gelfand, Frank, and Green that a tissue-bound receptor for activated C3 (C3b) is found in the normal human renal glomerular capillary wall. This receptor binds SRBC coated with IgM and anti-SRBC antibody and complement. The glomerular C receptor is specific for C3b, because EA coated with purified guinea pig or human C1, C14, C142 or C1423d are not bound. In this report we demonstrate the receptor for C3b located in the luminal surface of the capillary wall of the normal human renal glomerulus. We have also examined abnormal human kidneys harboring either immunologically or non-immunologically mediated lesions for the presence of C3b receptor, and the results were correlated with in vivo deposition of immunoglobulins and C3.
Hereditary C2 Deficiency: HLA Gene Complex Associations with Recombinant Events
Authors: Glass D. , Gibson D. J. , Carpenter C. B. , Schur P. H.
Published: December 2022

Ten families with genetically determined C2 deficiency have been studied for HLA gene complex associations. Nine of the propositi were heterozygous and one homozygous for the deficiency; all 10 also had rheumatic disease. C2 deficiency was found to be associated with the HLA-A10,B18 haplotype in seven families and with A2,B18; AW25,B18; and A10,BW40 haplotypes in the other three. Mixed lymphocyte culture (MLC) was used to define the HLA-D locus (MLC, mixed leukocyte reaction (MLR)) among C2-deficient individuals in order to obtain additional information as to the localization of the C2 gene on the HLA chromosome. A sibling pair, identical for the HLA-A and B loci, with the C2 heterozygous-deficient state linked to the A10,B18 haplotype in both, were mutually non-responsive in mixed lymphocyte cultures. Another sibling pair also identical for the A and B loci were, by contrast, mutually stimulatory in MLC. One sibling is C2 heterozygous deficient and the other is C2 normal.
Quantitative Requirements for C3 in Biologic Reactions: Forssman Cutaneous and Systemic Anaphylaxis, Reversed Passive Arthus Cutaneous Vasculitis, and Bactericidal and Opsonic Activity
Authors: Glovsky M. M. , Newman S. , Ghekiere L. , Alenty A.
Published: December 2022

In order to block the systemic Forssman anaphylactic reaction, guinea pig C3 activity had to be depleted by greater than 99% by using the C3 inactivator purified from cobra venom. When C3 functional hemolytic activity was greater than 200 units/ml (normal C3 activity = 20,000 to 160,000 units), guinea pigs uniformly died of anaphylaxis. Both the hemorrhagic reaction characteristic of Forssman cutaneous vasculitis and the reversed passive Arthus cutaneous vasculitis were suppressed when C3 activity was reduced to less than 1000 units/ml (approximately 1 to 3% of normal guinea pig C3 activity.) Using homozygous human C3-deficient serum from two separate patients, we studied bactericidal activity against E. coli 0111 and opsonic activity with Escherichia coli K-12, Staphylococcus aureus, and Staphylococcus albus. Serum bactericidal activity against E. coli 0111 was restored to normal with 50 units of human C3. A more pronounced opsonic defect was found with S. aureus and S. albus than with E. coli.
Cold Activation of the Classical Complement Pathway found in the Patients with Chronic Liver Diseases
Authors: Kitamura H. , Nagaki K. , Inai S.
Published: December 2022

Serum CH50 of 8 of 61 tested patients with chronic liver diseases were extremely low whereas CH50 of EDTA- or heparin-plasma were within the normal range. Serum and plasma complement profiles of these patients showed that hemolytic activities of C4 and C2 in serum were significantly lower than those in plasma.

When venous blood was obtained from the patient and clotted at different temperatures for 3 hr, C4 activity of serum separated at 37°C was as high as that of EDTA-plasma but C4 activity of the serum separated at 21°C or 4°C was extremely decreased. Also it was found that when patient serum was incubated with the serum of a normal subject, C4 activity in normal serum was decreased at 4°C, but the inactivation was inhibited in the presence of EDTA or heparin or when incubation was carried at 37°C. These results indicated that low serum CH50 found in some of our patients with chronic liver diseases might be due to the cold activation of the classical complement pathway in vitro.
C Dependent Immune Lysis of Virus Altered Plasma Membranes: Dependence on the Alternative C Pathway and Factor B
Authors: Perrin L. H. , Joseph B. S. , Oldstone M. B. A.
Published: December 2022

Antibody-mediated C dependent lysis of cell lines infected with herpes type 1 (HeLa cells), herpes type 2 (HeLa), influenza Å (Chang), measles (HeLa) and mumps (Vero) were analyzed for C participation. Lysis occurred with immune human sera, Mg++ EDTA immune sera, and immune sera depleted of C4 or treated with Fab anti-C4. Lysis did not occur with non-immune sera, Mg++EDTA sera, sera heated 50°C for 25 min, sera depleted of Factor B, or sera treated with Fab anti-Factor B. Lysis was restored to heated sera and Factor B immunodepleted sera by addition of Factor B but not by C2. Properdin and Factor D depleted immune sera lost significant lytic activity, but to a lesser extent than serum depleted of Factor B. Addition of depleted factors in physiologic concentrations partially restored the lytic activity.

Further studies showed that lysis of measles infected HeLa cells was induced by both immune IgG and F(ab')2 but not Fab' in the presence of a non-antibody containing human C source.
Reaction-Mechanisms Of C3NeF
Authors: Peters D. K. , Amos N. , Sissons J. G. P. , Lachmann P. J.
Published: December 2022

The cofactor requirements of C3NeF have been examined by using purified factors, C3, B, and D. Our experiments indicate that in the presence of NeF two C3-cleaving enzymes can be generated: when D is not added to the reaction mixture, B cleavage was not observed, whereas the addition of quantities of D sufficient to allow C3b feedback activation by purified C3b or CVF led to B cleavage. Both the B cleaving and the non-B cleaving reactions were inhibited (in a dose and time-dependent fashion) by preincubation of the C3 source with C3b inactivator (KAF) indicating that C3b rather than C3 is required. We envisage two enzymes (NeF) and (NeF), NeF functioning (as suggested by Fearon and Austen) to stabilize them. Thus, in the presence of NeF trace amounts of C3b—provided, for example, by spontaneous C3 breakdown or by other plasma proteolytic enzymes (“C3 tickover”)—would lead to the generation of an efficient C3 convertase.
Synthesis of C4 by C4 Deficient NIH-Albany Strain Guinea Pig Radiation Chimeras: Restoration of the Classical Pathway of Complement
Authors: Rudofsky U. H. , Webster R. O. , Pickering R. J.
Published: December 2022

We have detected significant hemolytic C4 activity after transplant of normal guinea pig bone marrow into lethally irradiated guinea pigs genetically deficient in C4. Each of five C4-deficient guinea pigs received 640 rads total body irradiation. They were immediately injected intracardially with 93 to 200 × 106 allogeneic nucleated marrow cells from Albany strain normal guinea pigs. The recipients were followed by sequential white cell counts and titrations of total complement, C1, C4, and C2. C4 activity, initially undetectable, was present as early as 24 days after transplantation and reached maximum levels by 45 to 60 days. C4 protein was also identified by using monospecific anti-guinea pig C4 serum. Amounts of C4 were sufficient to restore classical pathway function. C1 activity, initially 2 S.D. below normal, rose dramatically to normal or greater than normal levels. C2 activity also increased. These data demonstrate that allogeneic donor stem cells protected the lethally irradiated C4-deficient recipients and these allogeneic radiation chimeras subsequently synthesize circulating functional C4.
Complex Formation of Properdin Factor B with C3b, an Essential Step in the Generation of the C3b Dependent C3 Cleaving Enzyme of the Properdin System
Authors: Vogt W. , Dames W. , Schmidt G. , Dieminger L.
Published: December 2022

Earlier studies demonstrated that in the presence of Mg++ human factor B interacts with C3b as obtained by trypsin treatment of human C3, and only subsequent to this interaction can be cleaved and activated by Factor D̄. Mg++-dependent binding of highly purified B to C3b was demonstrated by the use of agarose to which trypsin-activated C3̄ had been fixed (Vogt et al., Z. Immunitaet forsch 149:440, 1975). Highly purified human C3b has now been prepared by treating partially purified C3hu with an insolubilized CVFB̄ complex, and fractionating the product on DEAE-cellulose and OH-apatite. The C3b preparation gave a single precipitation line in immunoelectrophoresis with anti-whole human serum, developed fusing precipitation bands with anti-C3c and anti-C3d sera in double immunodiffusion and did not react with antiserum against the B antigen of human C3. C3b and Mg++ were essential for cleavage of B by D̄ and for generation of C3-cleaving activity.
Characterization of Two Species of Fc Fragments Obtained from Rat IgG2a by Prolonged Papain Digestion
Authors: Arend William P. , Hermodson Mark A. , Teller David C.
Published: December 2022

Prolonged papain digestion of rat IgG2a produced two molecular species of Fc fragments, termed Fc(I) and Fc(II). Studies by gel filtration chromatography and polyacrylamide gel electrophoresis in SDS/urea indicated that the two subunit polypeptide chains in each Fc preparation were associated by non-covalent bonds only. By analytical ultracentrifugation Fc(I) was found to have a m.w. of 47,100 and a sedimentation coefficient of 4.08S. Fc(II) had a m.w. of 39,800 and a sedimentation coefficient of 3.83S. The m.w. for the subunit chains of Fc(I) and Fc(II) were 25,300 and 20,300, respectively, as determined by analytical ultracentrifugation under dissociating conditions. Calculation of the frictional coefficient ratios indicated that both Fc fragments possessed compact globular structures.

The difference in size between these two Fc fragments probably was due to a loss of some carboxy-terminal residues in Fc(II). Both Fc fragments possessed nearly identical amino-terminal amino acid sequences. Papain cleavage occurred primarily between residues 233/234 and 234/235. The carbohydrate compositions of the two species of Fc fragments were similar.

It was concluded that under acid and reducing conditions papain cleavage of rat IgG2a occurred to the carboxy-terminal side of the hinge region. Prolonged papain digestion led to secondary attack in the carboxy-terminal end of the Cγ3 domain at an unidentified site, or sites, producing a stable second species of Fc fragments.
Catabolism and Biologic Properties of Two Species of Rat IgG2a Fc Fragments
Authors: Arend William P. , Webster Douglas E.
Published: December 2022

Prolonged papain digestion of rat IgG2a has recently been shown to yield two species of Fc fragments, termed Fc(I) and Fc(II). The results of structural studies indicated that Fc(II) fragment was 15 to 20% smaller than Fc(I), probably secondary to loss of a carboxy-terminal peptide.

The effects of these structural alterations on the catabolism and biologic properties of the Fc fragments were determined. The results of catabolic experiments indicated that after injection into normal rats Fc(I) fragments were retained in the circulation and slowly catabolized whereas Fc(II) fragments rapidly underwent filtration in the kidneys. In nephrectomized rats, however, both Fc(I) and Fc(II) fragments possessed identical slow rates of catabolism, as determined by serum disappearance and whole body catabolic experiments. Fragment pFc′, corresponding to Cγ3 domain, was different from either of the Fc fragments in exhibiting rapid rates of catabolism in both normal and nephrectomized rats. Fc(I) was more active in complement fixation and in adherence to the Fc receptor on monocytes in comparison with Fc(II).

These results support the conclusion that catabolism of Fc and maintenance in circulation are separate processes influenced by different structures in the Fc fragment. The catabolism of Fc is controlled by structures in the Cγ2 domain. This process probably is not related either to complement fixation or to adherence to the Fc receptor on monocytes. Some correlations between the structure and biologic properties of these Fc fragments are discussed.
A New Histogenetic Method for Minor Histocompatibility Antigen Typing
Authors: Hauptfeld Věra , Klein Jan
Published: December 2022

A new method for typing minor H antigens is described. The method is based on the observation that effector T cells, sensitized in vivo to minor H antigens and then rechallenged in vitro, will lyse labeled target cells in the CML assay, provided that the target, the sensitizing, and the responding cells share the same allele at H-2K or H-2D loci. In this method, two congenic lines differing at a minor H locus are cross-immunized (by skin grafting, injection of lymphoid cells, or, preferably, by a combination of both treatments), rechallenged in vitro, and tested against a panel of strains carrying the same H-2 haplotype as the two lines. Target cells of the panel carrying the same minor H antigens as the strain against which the effector cells were sensitized are lysed; target cells not carrying these antigens are unaffected. The feasibility of the method was demonstrated by testing for H-3 antigens of a panel of H-2b-bearing strains. The new method is faster and more economical than the classicial F1 method which has been used until recently.
Bullous Pemphigoid Antigen: Isolation from Normal Human Skin
Authors: Diaz Luis A. , Calvanico Nickolas J. , Tomasi Thomas B. , Jordon Robert E.
Published: December 2022

A protein moiety from epidermal PBS-soluble products was isolated by gel filtration (Bio-Gel A-1.5m) and ion exchange chromatography (DEAE-cellulose). This protein (A-1-Epid) was not retarded by DEAE-cellulose in Tris-HCl buffer, 15mM, pH 8.1. By IEP against an antiserum to epidermal antigens, it showed a single cathodal arc. On disc electrophoresis, at low pH (4.3) a single band was apparent. On SDS gels this protein demonstrated two bands, one with a molecular weight of 20,000, and the second with a molecular weight of 9,200. This purified antigen was able to block the staining of the basement membrane zone produced by bullous pemphigoid antibodies on monkey esophagus and normal human skin with the use of indirect immunofluorescence. This study also demonstrates that bullous pemphigoid antigen (A-1-Epid) and a second epidermal protein (A-2-Epid) are present in the PBS-soluble products of human esophageal mucosa, saliva, and urine. These antigens appear to be unrelated with the blood group substances or secretor status of the donors.
Cyclophosphamide-Induced Specific Suppression of the Protective Immune Response to Rodent Malaria
Authors: Wells Robert A. , Diggs Carter L. , Phillips S. Michael
Published: December 2022

Nonspecific and specific chemosuppression of the immune response to Plasmodium berghei protective antigens were investigated. Specific immunosuppression was defined operationally as the selective suppression of the protective response to the parasite in mice injected with a combination of γ-irradiated infected mouse erythrocytes (γPb) and cyclophosphamide (CY) with continued responsiveness to sheep erythrocytes (SRBC). After initial treatment (γPb + CY), mice were injected with γPb in potentially immunogenic doses. These and appropriate control animals were later challenged with nonirradiated infected mouse erythrocytes. The influence of the initial treatment regimens on the protective response was evaluated by parasitemia, and mortality was observed after challenge. Specificity of suppression was measured by evaluating the ability of mice to produce antibody to SRBC.

Both specific and nonspecific suppression of the protective response to malaria were noted. Initial treatment with drug alone resulted in increased parasitemia and mortality and suppression of the SRBC antibody synthesis in drug-pretreated immunized mice as compared with immunized mice not pretreated with the drug. On the other hand, suppression of the response to the parasite, but not to SRBC, in animals pretreated with γPb + CY was clearly greater than that induced by drug alone. Thus, animals treated with malarial antigen and cyclophosphamide develop a measurable specific immunosuppression. These studies indicate that immunity to malaria is influenced by both cyclophosphamide alone (general immunosuppression) and cyclophosphamide in combination with antigen (specific immunosuppression) in a manner analogous to other immune responses.
Antibody-Mediated Suppression of Tumor Growth
Authors: Johnson Robert J. , Pasternack Gary R. , Shin Hyun S.
Published: December 2022

The effectors that cooperate with IgG1 in causing tumor suppression were identified as exudate macrophages and blood platelets in animals syngeneic and allogeneic to the tumor. Lymphocytes were not active in suppressing tumor growth in the presence of IgG1 antibody, although they were active with whole antiserum.