bluuebunny/crossref_metadata_2025 huggingface dataset:
Protection of Adult Mice against Tumor Challenge by Immunization with Irradiated Adult Skin or Embryo Cells
Authors: Bendich Aaron , Borenfreund Ellen , Stonehill Elliott H.
Published: December 2022

Water-soluble mouse embryo antigens, not detectable in normal adult visceral tissues, were found in each of 99 different cancerous tissues in 21 mouse strains (1, 2). Rabbit antiserum against the embryo constituents, absorbed with extracts of adult mouse internal organs, served as the diagnostic reagent in the agar double-diffusion precipitin tests which were employed for detection. Positive reactions were obtained not only with extracts of embryos and all cancers examined, but with extracts of the skin of adult animals as well.

Chromatographic estimation of the molecular weights of the antigens from embryos and tumors (65,000 to 70,000) revealed them to be about five times larger than the cross-reacting components in adult skin (10,000 to 15,000). The skin antigens were not detected in embryo viscera or in tumors, even in a sensitive radial immunodiffusion assay with the above antiserum (manuscript in preparation).
Physical and Chemical Studies on Human Properdin Purified by Elution from Zymosan and by Affinity Chromatography
Authors: Minta Joe O. , Lepow Irwin H.
Published: December 2022

Properdin purified to homogeneity from human serum after adsorption to and elution from zymosan (J. Immunol., 100: 142, 1968) is a glycoprotein containing about 9.8% carbohydrate. Preliminary analyses indicate 3.8% hexose, 1.5% hexosamine, 3.8% sialic acid, and 0.7% fucose. The V̄ calculated from amino acid composition and corrected for carbohydrate content was 0.70. Approach to equilibrium ultracentrifugation then yielded a molecular weight (m.w.)1 of 184,000 ± 12,000. In 6 M guanidine hydrochloride, properdin eluted from a 4% agarose column as a single protein peak with an estimated m.w. of 45,000, suggesting dissociation into four non-covalently linked subunits of similar or identical m.w. Since the possibility exists that zymosan-purified properdin may be in an altered state, properdin has been isolated by affinity chromatography. A column was charged with the IgG fraction of monospecific rabbit anti-human properdin which had been coupled to CNBr-activated Sepharose 4B (1:50 w/w) in 0.06 M barbital buffer, pH 9.6.
The C3 Feedback Cycle of C Activation: The Role of the C3b Inactivator (KAF)
Authors: Lachmann P. J. , Nicol P. A. E.
Published: December 2022

The immunochemical depletion of KAF from serum has been brought about by using purified F(ab')2 anti-KAF. The depleted serum undergoes a spontaneous activation of its alternate pathway, converting its GBG and depleting the co-factors required to form the “cobra venom C3 convertase.” All C3 becomes converted to C3b and if red cells are present C3 becomes fixed to them. This situation exactly parallels that seen in vivo in the unique KAF-deficient patient. The mechanism by which KAF depletion leads to activation of the alternate pathway has been shown to lie in its failure to activate C3b, since if C3 is first depleted from serum with F(ab')2 anti-C3 subsequent KAF depletion produces no alternate pathway activation. In serum depleted of C3 it is similarly found that neither zymosan, endotoxin nor inulin can induce the formation of a C3 convertase. It is therefore clear that the alternate pathway is in reality a C3-feedback pathway.
Isolation of Human C4 by Affinity Chromatography with IgG-Coupled Sepharose 6B
Authors: Patrick R. A. , Mernitz Janice , Bing D. H.
Published: December 2022

At least four examples exist which demonstrate an association of C4 with serum proteins. It has been previously reported that C4 forms association products with itself and with human 7S γ-globulin in the presence of C1s̄. Native C4 is known to complex with C2 in a reversible fashion. This reversible association of C2 and C4 is converted to a firm complex which is due to the proteolytic action of C1s̄. Finally, C4 is believed to react with receptor sites on antibody molecules as well as erythrocyte membrane receptors during immune hemolysis.

Human IgG was covalently linked to Sepharose 6B under basic conditions by use of cyanogen bromide. The resultant Sepharosecoupled IgG was stored at 4°C with 0.005% sodium azide until ready for use. Approximately 1 mg of IgG was bound per 20 mg Sepharose. Crude C4 from a pseudoglobulin fraction of human serum and subsequently eluted from DEAE-cellulose at pH 7.3 was used for studies with insoluble IgG.
Hereditary C6 Deficiency in Man
Authors: Leddy J. P. , Frank M. M. , Gaither T. , Heusinkveld R. S. , Breckenridge R. T. , Klemperer M. R.
Published: December 2022

An 18-year-old black female (D.B.) in good general health was found to have no hemolytic activity in repeated serum CH50 titrations. Functional assays yielded normal values for all C components except C6. C6 was not detectable a) by functional assays utilizing either stepwise buildup of C components on EA or reconstitution of C6-deficient rabbit serum, or b) by immunodiffusion against antiserum to rabbit C6. Hemolytic activity was restored by addition of functionally pure C6. Hemolytic C (CH50) titers on family members were normal. By specific functional assay, however, both parents and five of six available siblings had approximately half-normal serum C6 levels. The other sibling was normal. C6 deficiency in the proband could not be accounted for by a serum inhibitor; C6 was also absent in D.B. plasma. Biologic properties of D.B. serum include: a) absent bactericidal activity against Salmonella typhi 0 901 with or without added rabbit antibody; b) normal generation of chemotactic activity for human neutrophils in the presence of endotoxin or aggregated IgG; c) ability to sensitize appropriate cells for immune adherence or agglutination by anti-C3 Coombs serum; and d) inability to lyse PNH red cells in either acid hemolysis or “sugar water” tests.
The Chemotactic Factor Inactivator in Normal Human Serum
Authors: Till G. , Ward P. A.
Published: December 2022

Normal human serum contains a chemotactic factor inactivator (CFI) that irreversibly inhibits the neutrophil chemotactic activity associated with fragments of C3 and C5, the C complex, and the bacterial chemotactic factor derived from Escherichia coli. Physicochemical features of CFI include the following: it is present in the pseudoglobulin fraction of serum, soluble in ammonium sulfate at 60% saturation, and elutes in at least two different zones in ion exchange chromatography and in Sephadex gel filtration. Electrophoretically CFI appears in positions of α- and β-globulins. CFI reacts in an enzymatic-like manner: the inactivation of chemotactic activity is progressive with time, is temperature-dependent, and is maximal at pH 7.3. CFI is heat labile, being destroyed at 56°C after ½ hr. Its activity is not blocked by 0.02 M EDTA, 0.02 M phenanthroline, soybean trypsin inhibitor, DFP or ε-aminocaproic acid. Caseinolytic activity and carboxypeptidase-B-like activity (measured by hydrolysis of hippuryl-l-arginine) are lacking in CFI preparations.
Receptors for Bound C3 on Neutrophil Membrane Fragments
Authors: Henson Peter M.
Published: December 2022

Neutrophils from rabbit and man have the property of adhering to particles (e.g., erythrocytes or zymosan) to which C3 is bound. Phagocytosis and release of granule constituents may follow. To study the receptors involved, we obtained plasma membrane fragments from these cells. A purified population of neutrophils was trace labeled with 125I anti-neutrophil antibody, disrupted in sucrose, and the nuclear debris, granules and soluble proteins were separated from the plasma membranes on sucrose gradients. Membrane fragments were labeled with 125I with lactoperoxidase and H2O2 and washed with saline. The fragments were examined by electron microscopy. Labeled membrane fragments from rabbit and human neutrophils were incubated with erythrocytes or zymosan, previously coated with C, which were then washed and counted. The fragments bound to EAC1423 but not to EAC142. Control preparations of rabbit erythrocyte membranes did not bind to EAC1423. The binding was partly temperature dependent and that of rabbit neutrophil membranes was inhibited by EDTA.
Isolation of the Seventh Component of the C System from Human Serum
Authors: Arroyave Carlos M. , Müller-Eberhard Hans J.
Published: December 2022

Isolation of C7 from human serum was carried out by a combination of Bio-Rex 70, DEAE-32, DEAE and CM 32 chromatography and preparative Pevikon block electrophoresis. The yield in terms of hemolytic activity was 0.5 to 1.5%. Purification was 1030-fold. The isolation and radiolabeled protein was identified by disc electrophoresis as a single protein and the C7 hemolytic activity could be localized to the position of this band. By two different methods the m.w. was determined to be 120,000. By immunoelectrophoresis the protein was found to be a slowly migrating β-globulin. The concentration in whole human serum was quantitated by the Mancini method to be 47 to 61 µg/ml. The efficiency of binding to sensitized sheep erythrocytes was found to be 5- to 10-fold greater when the protein was offered to EAC1-3 cells in presence of C5 and C6, rather than to EAC1-6 cells alone. Complete proportionality was obtained between -ln(1-y) and the dose of C7 protein employed.
The Terminal Components of the Nurse Shark (Ginglymostoma cirratum) C System
Authors: Jensen Joerg A. , Fuller Laphalle , Iglesias Elena
Published: December 2022

The preterminal (t1) and terminal (t2) C-components were isolated from nurse shark serum in functionally pure form. Preparative procedures included: a) low ionic strength precipitation (µ = 0.006, pH 7.6) where the activities stayed in the supernatant; b) DE-52; c) hydroxyapatite column chromatography; and d) gel filtration (G-200). In step (b) t1 eluted before t2 (µ = 0.006 and 0.06 at pH 7.6, respectively). In step (c) the order was reversed, t2 eluted with 0.2 M PO4, t1 with 0.5 M PO4 at pH 6.8. Functional purity was obtained after step (c). On a calibrated Sephadex G-200 column both components eluted with an IgG marker. The m.w. of t2 was determined by polyacrylamide gel electrophoresis; it was 185,000.

Both components were assayed hemolytically in two systems: 1) with a stable intermediate complex consisting of sheep E optimally sensitized with natural shark antibody and reacted sequentially with purified C1n and two shark serum fractions containing at least two still undefined C-components; and 2) with EAC1gpC4-C7hu.
Cerebrospinal Fluid C
Authors: Propp Richard P.
Published: December 2022

Determination of C3 concentration in unconcentrated cerebrospinal fluid by electroimmunodiffusion was possible only when an immunofixation layer was added by using antibody to the original gel antibody against C3. Recently we have been able to measure C4 in this manner, but in this situation the original running time is critical and must not be excessive since then the precipitate dissolves. This technique was applied to the study of paired sera and cerebrospinal fluids. Native C3 and C3 conversion products may be visualized by antigen-antibody crossed electrophoresis after concentration of cerebrospinal fluid 40-fold by ultrafiltration, providing the immunofixation procedure is used.

Ten normal specimens of cerebrospinal fluid had a mean (± 1 S.D.) of 378 ± 159 µg/100 ml for C3, with a mean C3/albumin of 1.49 ± 0.18%. Eight normal spinal fluids had a mean for C4 of 274, with a mean C4/albumin of 1.23%.
Initiation of Oxidative Metabolism in Human Neutrophils (PMN) by C Activated through an Alternate Pathway
Authors: Strauss Ronald G. , Mauer Alvin M. , Spitzer Roger E. , Asbrock Theresa , Stitzel Ann E.
Published: December 2022

An oxidative metabolic burst comparable to that after phagocytosis was initiated in normal PMN by exposing them to sera incubated with powdered inulin. In these sera a change in the electrophoretic mobility of the C3PA from beta to gamma indicated activation of the alternate pathway, while consumption of C3 indicated C activation. An increase of hexose monophosphate shunt activity in PMN greater than 5-fold indicated stimulation. Increases in O2 consumption, NBT dye reduction, and degranulation confirmed the metabolic burst. PMN metabolism was stimulated by inulin in fresh sera and in sera depleted of C4 from patients with active hereditary angioneurotic edema. Stimulation did not occur in the absence of serum, in sera heated to 50°C or 56°C for 30 min, in 0.02 M EDTA serum, in sera depleted of C3 or C3PA by adsorption at equivalence with monospecific antisera. The chemotactic response of these stimulated PMN was defective, suggesting altered membrane function.
Lysosomal Enzyme Release from Human Leukocytes: Mediation by the Alternate Pathway of Complement Activation
Authors: Goldstein Ira M. , Brai Melchiorre , Osler Abraham G. , Weissmann Gerald
Published: December 2022

The alternate pathway of complement activation has recently assumed an important role in the mediation of inflammation and tissue injury. We have found that complement activated via this pathway interacts with human polymorphonuclear leukocytes (PMN) in the absence of particulates and stimulates the selective release of lysosomal enzymes. Fresh human serum, after treatment with zymosan or cobra venom factor, yields a fluid phase component which induces cytochalasin B-treated PMN to release β-glucuronidase without leakage of cytoplasmic lactate dehydrogenase. Involvement of the alternate pathway of complement activation is evidenced by failure of serum to yield enzyme-releasing activity after pretreatment with rabbit antihuman C3 proactivator, 0.01 M EDTA but not 0.01 M EDTA, 0.02 M salicylaldoxime, 0.02 M hydrazine or by heating to 56°C for 30 min. Levels of C3 proactivator in treated sera correlated inversely with and perturbs the PMN plasma membrane sufficiently to cause lysosomal membrane perturbation, fusion, and ultimately lysosomal enzyme extrusion by a process of “reverse endocytosis.”
Effects of Pyran Copolymer on Leukemogenesis in Immunosuppressed AKR Mice
Authors: Hirsch Martin S. , Black Paul H. , Wood Mary L. , Monaco Anthony P.
Published: December 2022

Treatment with anti-lymphocyte serum twice weekly from 4 months of age accelerated leukemogenesis in AKR mice. In contrast, monthly injections of Pyran copolymer retarded development of leukemia both in immunologically intact and suppressed animals of this strain. These studies provide further evidence for the absence of immunologic tolerance to viral or tumor antigens in AKR mice, and for the efficacy of Pyran copolymer as a prophylactic agent against virus-associated oncogenesis in immunosuppressed hosts.
Myelination Inhibition Factor: Its Absence in Sera from Subhuman Primates Sensitized with Myelin Basic Protein
Authors: Seil Fredrick J. , Rauch Helene C. , Einstein Elizabeth Roboz , Hamilton Ann E.
Published: December 2022

Experimental allergic encephalomyelitis was induced in subhuman primates by inoculation with either whole monkey brain plus Freund's adjuvant or with purified bovine myelin basic protein combined with Freund's adjuvant. Sera from the whole brain-sensitized primates, when incorporated into tissue culture nutrient medium and applied continuously to cerebellar cultures from the day of explantation on, prevented the formation of myelin in the explants. Sera from basic protein-sensitized primates, on the other hand, failed to inhibit myelination when applied in like fashion to cerebellar tissue cultures, confirming a previous demonstration of a similar lack of myelination-inhibiting activity in sera from basic protein-sensitized guinea pigs. The results suggest that the tissue culture myelination inhibition factor is induced by a component of whole central nervous system tissue other than the encephalitogenic protein.
Complement As a Regulator of Interactions Between Immune Complexes and Cell Membranes
Authors: Miller Gary W. , Nussenzweig Victor
Published: December 2022

Soluble immune complexes prepared with limited amounts of complement and injected into mice bind to circulating cells, and are then rapidly released into the plasma. The complex release is complement-dependent. Complexes prepared with excess complement do not bind to the circulating cells or to mouse lymphocytes in vitro; thus the target of the complement activity is the complex, not the cell membrane. Since complement can act to either increase or decrease the affinity of immune complexes for cell membranes, depending upon various factors, we propose that complement serves as a sensitive regulator of interactions between immune complexes and cell membranes.
Characterization of Antibodies Specific for Polyglycerophosphate
Authors: Frederick G. Thomas , Chorpenning Frank W.
Published: December 2022

The immunoglobulin classes of antibodies specific for polyglycerophosphate (PGP) in human, guinea pig, rabbit and rat sera detected by passive hemagglutination of antigen-coated erythrocytes were characterized by column chromatography, ion-exchange chromatography, 2-mercaptoethanol sensitivity, and precipitation with monospecific antisera. All serum samples from normal (uninjected) humans and guinea pigs had anti-PGP antibodies in both the IgM and IgG classes. Normal rats were divided between those with antibodies in both classes and those with antibodies in the IgM class only. All normal rabbits had antibody in the IgM class only and produced antibody in both IgM and IgG only after injection (i.p.) of antigen. It is suggested that the results in rabbits may be due to presentation of antigen to different immunocompetent cell populations or an involvement of antigen-specific T cells in the production of the IgG antibody. All guinea pig IgG anti-PGP antibodies, whether from normal or antigen-injected animals, were restricted to the IgG1 subclass. Guinea pigs injected with certain other antigens produced antibodies in both subclasses. A hypothesis is presented to account for this difference.
Combined Detection of Human T and B Lymphocytes by Rosette Formation with Sheep Erythrocytes and Zymosan-C3 Complexes
Authors: Mendes Nelson F. , Miki Silvia S. , Peixinho Zulma F.
Published: December 2022

A new method for detecting human complement receptor (B) lymphocytes by utilizing rosette formation with zymosan particles treated with mouse complement (ZC) has been described in this paper. The method has been compared with rosette formation with human erythrocytes sensitized with rabbit antibody and mouse complement (HEAC). T cells have been detected by rosette formation with sheep erythrocytes (E). The combined detection of both B and T lymphocytes could be performed by using as indicator a mixture of ZC and E. Mean percentage values for E and ZC were 60.5 and 15.2 by using the combined determination to study lymphocytes from 30 normal subjects. The mean percentage value for HEAC was 19.5. Only a small proportion of lymphocytes (1 to 2%) formed mixed rosettes with ZC and E. This paper further demonstrates the use of this combined indicator for the study of the distribution of B and T cells in tissue sections.
The Inactivation of Human C1 by Benzamidine and Pyridinium Sulfonylfluorides
Authors: Bing David H. , Cory Michael , Doll Michael
Published: December 2022

Affinity chromatographically purified C1̄ was demonstrated to be inhibitable by a variety of benzamidine and pyridinium fluorosulfonyl-substituted compounds. Four of the compounds tested afforded greater than 50% inhibition at concentrations between 10-6 and 10-5 M. The data indicated that inhibition was greatest with those compounds which contained a substituted benzamidine and sulfonyl-fluoride group. The site of action was the esterase activity associated with the C1.
Human Lymphocyte Membrane Immunoglobulin as a Reflection of Maturation
Authors: James Karen K. , Hurtubise Paul E. , Macpherson Colin R. , Murphy Samuel G.
Published: December 2022

Peripheral blood lymphocytes can be divided into two subpopulations on the basis of surface antigens and membrane receptors. It is generally well recognized that lymphocytes that contain readily detectable surface or membrane-bound immunoglobulin (M-Ig)3 are B lymphocytes (1). The published data on the normal range of B lymphocytes in various studies have not taken into account the possibility of age-related changes in the number and class distribution of human B lymphocytes (2–12). This study was designed to investigate both the number and class distribution of M-Ig on human B lymphocytes obtained from normal subjects in the 1st through the 8th decade of life.

The following points are demonstrated: 1) Children (ages 1 to 10) have B lymphocytes bearing IgM (M-IgM) in significantly greater numbers than adults ages 21 to 90 (t = 6.03, p < 0.001). 2) Lymphocytes bearing IgG (M-IgG) in children appear decreased when compared to adults (t = 2.37, p < 0.02).
Separation of T and B Lymphocytes by Nylon Wool Columns: Evaluation of Efficacy by Functional Assays in Vivo
Authors: Trizio Domenico , Cudkowicz Gustavo
Published: December 2022

A study was made of antibody-forming cell precursors (B cells) and thymus-derived helper cells (T cells) of mouse spleen filtered through nylon wool columns. Two populations of spleen cells were separated: effluent cells nonadhering to the nylon fibers, and nylon-adherent cells which could be eluted from the columns by mechanical agitation. The presence or absence of functional lymphocytes was demonstrated by transplantation into irradiated syngeneic recipients, challenge with antigen, and assay for specific antibody-forming cells. By transplanting each of the column-filtered subpopulations separately, in combination, and with either bone marrow or thymus cells (known sources of B or T lymphocytes, respectively), it was possible to establish that B and T cells were separated by the column filtration; the effluent population was enriched with functional helper cells and depleted of precursor cells, whereas the nylon-adherent population was enriched with competent precursors and depleted of helpers.
Type-Specific Antibodies in Human Chlamydia Trachomatis Infections of the Eye
Authors: Briones Odeon C. , Hanna Lavelle , Jawetz Ernest , Dawson Chandler R. , Ostler H. Bruce
Published: December 2022

Sera from humans with ocular chlamydial infections were examined for type-specific antibodies by immunofluorescence before and after extensive cross-absorption with type-specific antigens of Chlamydia trachomatis. In trachoma-inclusion conjunctivitis infections ascribable to types C, A, or F, and occasionally types B and E, the antibody response was so narrow and specific as to make serologic diagnosis of the infecting agent feasible. In other ocular or oculongenital infections with types B, Ba, E, and D, and with lymphogranuloma venereum, there was only a broad antibody pattern. In experimental infections with types F or D, the initial antibody response of volunteers was narrowly directed at the dominant type-specific antigen of the infecting strain; then it broadened with time to include cross-reactive antibodies. In sequential infections with different types, the antibody to the first infecting type was recalled to a higher level by the second, heterotypic infection. In some individuals, treatment with tetracycline drugs resulted both in regression of clinical activity and decrease in antibody titer. Perhaps such a serologic response may indicate loss of antigen and cure.
Studies on Monoclonal Antibodies
Authors: Stone Marvin J. , Fedak Jennifer E.
Published: December 2022

Factors influencing the precipitation of a mixed cryoglobulin formed by the interaction of a Waldenström macroglobulin antibody (IgMSie) with its antigen (human IgG) have been studied. Binding of FabµSie fragments to a G1, κ myeloma protein in the analytical ultracentrifuge showed that the IgG antigen was effectively univalent. Consequently, IgM(IgG)10 is the largest potential complex that can form in this system. Experiments performed by the method of equilibrium molecular sieving demonstrated that primary affinity of antibody combining site for antigenic determinant exhibits modest temperature dependence similar to that observed in other antigen-antibody systems. Solvent ionic strength and change in pH near neutrality did not significantly alter the association constant. By contrast, precipitation of the IgMSie-IgG complex was profoundly affected by variation in temperature, ionic strength, and pH. Larger complexes were required for precipitation to occur when the reaction was carried out under non-optimal environmental conditions or when certain IgG antigens were employed. These results indicate that the cryoprecipitating phenomenon is explicable by the finite size and limited solubility of the IgM-IgG immune complexes.
Resistance to Tumor Growth Mediated by Listeria Monocytogenes. Destruction of Experimental Malignant Melanoma by Lm-Activated Peritoneal and Lymphoid Cells
Authors: Youdim Said
Published: December 2022

A murine experimental model of nonspecific tumor destruction mediated by cells activated by Listeria monocytogenes (LM) is described. B16 melanoma growth is prevented or suppressed in the syngeneic host when tumor cells are inoculated in contact with viable LM. In vitro, cultured B16 cells are destroyed by LM immune peritoneal or splenic cells in the presence of the bacterial antigen(s). Activation of LM immune cells in vitro is immunologically specific. Replacement of LM by sheep red blood cells or bovine serum albumin in the in vitro cultures aborts the cytotoxic effect. Further, no tumor cell killing is obtained when thioglycollate-induced or normal peritoneal cells are substituted for LM immune cells in the in vitro cultures. Normal spleen cells in the presence of LM are weakly cytotoxic for B16 cells. Normal peritoneal cells plus LM or LM alone are not. Elimination of thymus derived “T” cells by anti-θ C3H or rabbit anti-mouse brain serum (RAMB) abrogated the cytotoxic effect. Therefore, LM-induced tumor destruction probably occurs through nonspecific mechanism(s) consequent to activation of host “T” cells by specific immune reactivity to LM antigen(s).
Antibody in the Sera of Tumor-Bearing Mice that Mediates Spleen Cell Cytotoxicity toward the Autologous Tumor
Authors: Blair Phyllis B. , Lane Mary-Anne , Mar Pearl
Published: December 2022

Pretreatment of MTV-induced BALB/cfC3H mammary tumor cells with autologous serum results in increased spleen cell cytotoxic activity and the recruitment of previously inactive spleen cells to cytotoxic activity against the target cells. These recruiting antibodies are tumor-specific for individual tumors; pretreatment with such serum of target cells of an MTV-induced mammary tumor obtained from a different BALB/cfC3H female results in blocking of spleen cell activity. The autologous recruiting factors are active at dilutions of 1000 or more of whole serum and are found in the 19S fraction after gel filtration.
Effect of Lectins on the Levels of cAMP and cGMP in Guinea Pig Lymphocytes: Early Responses of Lymph Node Cells to Mitogenic and Non-Mitogenic Lectins
Authors: Burleson David G. , Sage Harvey J.
Published: December 2022

Guinea pig lymph node lymphocytes were reacted with two mitogenic lectins (LcL-A or PHA) and two nonmitogenic lectins (AbL or WGA), and the levels of cAMP and cGMP were determined during 30 min of culture. Various concentrations of LcL-A and PHA caused modest (1.5 to 2-fold) rises in cAMP levels. However, the rise in cAMP did not correlate with the degree of mitogenicity. AbL caused a greater rise in cAMP than the mitogenic lectins whereas WGA had no effect on cAMP levels. None of the four lectins causes an appreciable (i.e., greater than 1.5-fold) change in cGMP levels. There does not seem to be any positive correlation between early changes of cAMP and/or cGMP and mitogenesis.
Adjuvant Regulation of T Cell Function
Authors: Reinisch Carol L. , Gleiner Nancy A. , Schlossman Stuart F.
Published: December 2022

The effect of complete Freund's adjuvant (CFA) on distinct T cell functions was investigated. Adjuvant was found to suppres the generation of cytolytic T cells in vivo when mixed with allogeneic P815 cells before immunization of C57BL/6 mice. Inoculation of the mice with either adjuvant or adjuvant emulsified with allogeneic cells resulted in whole splenic populations or immunoabsorbent-purified T cells that did not generate cytolytic activity in vitro against allogeneic cells. Mixing T cells from normal and adjuvant-treated mice before in vitro sensitization resulted in suppression of lytic activity. However, memory T cells were not subject to the same suppressive regulation as were precytotoxic T cells since adjuvant had no effect on subsequent boosting of memory.

Adjuvant augmented the humoral response to allogeneic cells since immunization of mice with P815 cells in adjuvant resulted in prolonged cytotoxic antibody levels. This response was predominantly IgG, reflecting the enhanced activity of another T cell subpopulation, helper T cells.

We conclude that the effect of adjuvant is compartmentalized among T cell subpopulations. Activity of precytotoxic T cells is diminished by suppressor cells, whereas helper T cell function is augmented. Memory T cells do not appear to be under the same regulatory control as precytotoxic cells since adjuvant does not suppress the boosting of memory T cell responses in vitro.
Generation of Cytotoxic T Lymphocytes in Vitro
Authors: Fitch Frank W. , Engers Howard D. , Cerottini Jean-Charles , Bruner K. Theodor
Published: December 2022

Irradiated cells obtained from MLC at the peak of the CTL response caused profound suppression of generation of CTL when added in small numbers at the initiation of primary MLC prepared with normal spleen cells. The inhibitory activity of the MLC cells was not affected by irradiation (1000 rads) but was abolished by treatment with anti-θ serum and complement. The suppression was immunologically specific. The response of A (H-2a) spleen cells toward C3H (H-2k) alloantigens was suppressed by irradiated MLC cells obtained from MLC prepared with A spleen cells and irradiated C3H-stimulating cells, whereas the response of A spleen cells toward DBA/2 (H-2d) alloantigens was affected relatively little. However, if irradiated C3H × DBA/2 F1 hybrid spleen cells were used to stimulate A spleen cells in MLC, addition of irradiated MLC cells having cytotoxic activity toward C3H antigens abolished the response to both C3H and DBA/2 antigens. The response to DBA/2 antigens was much less affected when a mixture of irradiated C3H and DBA/2 spleen cells was used as stimulating cells. Thus, the presence of MLC cells having cytotoxic activity toward one alloantigen abolished the response to another non-cross-reacting antigen only when both antigens were present on the same F1 hybridstimulating cells. This suppression of generation of CTL by irradiated MLC cells apparently involves inactivation of alloantigen-bearing stimulating cells as a result of residual cytotoxic activity of the irradiated MLC cells. This mechanism may be active during the decline in CTL activity noted in the normal immune response in vivo and in vitro.
Humoral Immunity in Aged Mice
Authors: Segre Diego , Segre Mariangela
Published: December 2022

Mixed cultures of DNP-primed spleen cells from old and young BC3F1 mice in diffusion chambers yielded significantly fewer secondary PFC than expected from the numbers of PFC found when young and old spleen cells were cultured alone. There was a negative correlation between the log of the number of PFC found in cultures of old spleen cells alone and the degree of immunosuppression exerted by the cells. The immunosuppressive effect was abolished by pretreatment of the old spleen cells with anti-θ serum and complement. Thus, the effect was attributed to the presence of large numbers of suppressor T cells in the spleen of primed old mice. No suppression was found in mixed cultures of spleen cells from old unimmunized mice and young primed mice.
Genetic Control of Immune Response against Random Copolymers of Glutamic Acid and Alanine (GA) and Tyrosine (GT) in Inbred Mice
Authors: Merryman Carmen F. , Muarer Paul H.
Published: December 2022

An immune response (Ir) gene(s) controlling responses to synthetic random copolymers of two amino acids of glutamic acid and alanine (GA) in mice has been well documented. This specific Ir gene is linked to the major histocompatibility complex of the species. Mice of the H-2a, b, d, f, k, r and s haplotypes produce high titer antibodies and thus are high responders; whereas mice of the H-2 haplotypes j, ja, p and q produce no detectable antibody and are nonresponders. The response is “all or none” and high response is dominant.

Attempts to detect an Ir-GT gene (glutamic acid and tyrosine) after immunization with two GT random copolymers were unsuccessful. However, GT antibody could be produced after immunization with GT-methylated bovine serum albumin aggregates.
Visual Detection of Carcinoembryonic Antigen on Surfaces
Authors: Giaever Ivar
Published: December 2022

An attempt is made to detect carcinoembryonic antigen (CEA) at clinically interesting concentrations by using a simple immunologic surface test. Antibodies to CEA are detected in a direct test at concentrations below 1 ng/ml. The sensitivity of this assay is mainly limited by diffusion to the reacting surface. CEA is detected in an inhibition test at concentrations down to 20 ng/ml. The sensitivity of this inhibition test is limited by the average equilibrium constant in solution of the antibody-antigen reaction.
An IgG-Fc Receptor Induced in Cytomegalovirus-Infected Human Fibroblasts
Authors: Keller Robert , Peitchel Ricki , Goldman John N. , Goldman Margaret
Published: December 2022

Cytomegalovirus- (CMV) infected cultured human fibroblasts incubated with normal human serum were shown to react with IgG by direct and indirect fluorescent antibody tests. The binding of IgG to infected cells was unrelated to the presence of anti-CMV complement fixing (CF) antibodies and was not observed with uninfected cells. The reaction was first observed 36 hr after infection as diffuse cytoplasmic staining which by 72 hr became localized into a dense perinuclear structure. The cytoplasmic fluorescence was not detected with anti-IgA or anti-IgM fluorescent conjugates. The reaction was seen with purified IgG and Fc fragments but was only minimally detectable with Fab fragments. It occurred in fibroblasts infected with three standard laboratory strains of CMV and seven recent CMV isolates from patients and was observed in six separate lots of human foreskin fibroblast cultures as well as in WI-38 cells. We conclude that CMV infection induces the formation of a IgG receptor in human fibroblasts.
A New Alloantigenic System Associated with the Mls Locus in the Mouse
Authors: Tonkonogy Susan L. , Winn Henry J.
Published: December 2022

An antiserum prepared by injecting C3H/HeJ mice with CBA/J tissues has been shown to react with cell surface components that are not part of any previously described system of serologically detectable alloantigens. The antiserum, which is designated AST-101, acts selectively in cytotoxic tests carried out with lymphoid cells, killing B cells, but not T cells. Phagocytic cells found in peritoneal exudates are also killed by AST-101 and complement in vitro; the sensitivity of other cell types has not been determined. Strain distribution does not indicate any association of the AST-101 system with H-2, Ly, or Thy systems; genetic analysis reveals close linkage with the mouse minor MLC-stimulating (Mls) locus. Serologic analysis also points to a close association between antigens reactive with AST-101 and the products of the Mls genes.
Factors Influencing the Isolation of Membrane Immunoglobulins from T and B Lymphocytes
Authors: Cone Robert E.
Published: December 2022

The optimal conditions for the extraction and isolation of immunoglobulins from the cell membrane (M-Ig) of T and B lymphocytes radioiodinated with lactoperoxidase were studied. A combination of 10 M-urea, 1.5 M acetic acid and the non-ionic detergents Triton X-100 or Nonidet P-40 (NP-40) was found to overcome the tendency for membrane proteins to precipitate after extraction in urea-acetic acid alone. However, detection of T cell Ig was more dependent on detergent concentration than B cell M-Ig. Less T cell Ig was detected when the detergent concentration was increased from 0.1% to 1.0%, whereas the amount of B cell M-Ig was not affected by higher detergent concentrations. All polyvalent and anti-κ-chain antisera tested reacted with T and B cell M-Ig, however, anti-µ-chain sera bound B cell M-Ig only. Disc electrophoresis (PAGE) of reduced and alkylated T M-Ig revealed polypeptide chains with mobilities of Ig µ and L chains and additional polypeptides with apparent m.w. of 45,000 and 30,000. The latter two polypeptides were present in nonspecific precipitates also. PAGE analysis of B cell M-Ig revealed µ-chains, a second heavy chain with an apparent m.w. of 65,000 as well as L chains, and, in addition, polypeptides with apparent m.w. of 45,000 and 30,000.

It is suggested that T cell immunoglobulin is structurally distinct from B cell membrane Ig and may share some properties characteristic of integral membrane proteins.
Macrophage Requirement for the in Vitro Response to TNP Ficoll: A Thymic Independent Antigen
Authors: Chused Thomas M. , Kassan Stuart S. , Mosier Donald E.
Published: December 2022

The in vitro antibody-forming cell response of mouse spleen cells to the thymic independent synthetic polymer, TNP-Ficoll, was found to require the presence of either macrophages or 2-mercaptoethanol. The TNP-Ficoll response and that to the thymic dependent antigen, sheep erythrocytes, demonstrated the same degree of macrophage dependence. However, macrophage-depleted spleen cells which did not produce plaque-forming cells responded normally to T and B cell mitogens. Comparison of the antibody-forming cell response of macrophage-depleted spleen cells in the presence and absence of 2-mercaptoethanol indicated that either macrophages function 10 to 100 times more efficiently in its presence or that 2-mercaptoethanol partially replaces the function of macrophages.
The Effect of Corticosteroids on Human IgG Synthesis
Authors: McMillan R. , Longmire R. , Yelenosky R.
Published: December 2022

The effect of prolonged, high-dose corticosteroid therapy on total IgG synthesis rates by human bone marrow and splenic leukocytes was studied in patients with immune thrombocytopenia. Marrow IgG production rates begin to decrease 3 weeks after beginning therapy and reach levels approximately one-fourth of pre-treatment rates after 6 weeks. No effect of corticosteroids on splenic IgG production rates could be shown. Marrow IgG synthesis rates in these patients were on the average from 3 to 10 times greater than corresponding splenic production rates and, in addition, appeared to correlate with serum IgG levels. These data suggest that the marrow is an important contributor to the total body IgG pool; and since corticosteroids appear to suppress marrow IgG production, this may be one reason for their therapeutic usefulness in antibodymediated diseases.
Antibodies to the Triplet Codons AAA, AAC, and AUG: Reactions with Nucleic Acids
Authors: D'Alisa Rose , Erlanger Bernard F.
Published: December 2022

Antibodies specific for the trinucleotide codons AAA, AUG, and AAC were examined for their reactions with nucleic acids. Anti-AUG and anti-AAC precipitated denatured DNA. Anti-AAA did not, and moreover, the binding of a tritiated AAA derivative to anti-AAA was not inhibited by denatured DNA. Radioligand-binding studies showed that anti-AAA was highly specific for the triplet sequence, some cross-reactions occurring with di-A and tetra-A but little with A and poly(A). The anti-codons did not precipitate whole yeast RNA, but binding could be demonstrated with Escherichia coli 3H-rRNA. Anti-AAC and anti-AUG (but not anti-AAA) bound E. coli 14C-tRNA. Anti-AUG was found to inhibit protein synthesis in an in vitro system in which rabbit reticulocyte mRNA was being translated. Inhibition was abolished by absorption of the antibody with AUG-RSA. The results of these and previous experiments (reference 1) are interpreted to indicate that antibodies can recognize trinucleotide sequences but not with absolute specificity, and it is suggested that antibodies to longer nucleotide sequences might show specificity comparable to that shown by antibodies to polysaccharide and peptide sequences.
Transfer of Hypogammaglobulinemia in two Inbred Chicken Strains by Spleen Cells from Bursectomized Donors
Authors: Palladino M. A. , Lerman S. P. , Thorbecke G. J.
Published: December 2022

Transfer of lymphoid cells from agammaglobulinemic donor chickens into 3- to 4-week-old irradiated recipients that had been surgically bursectomized at 2 to 3 weeks of age significantly depressed the ability of the antibody-forming apparatus to recover from irradiation. Antibody production to Brucella abortus and sheep erythrocytes remained much below control levels in BX donor-cell recipients of both chicken strains studied (SC and FP). Progressive loss of serum IgM and IgG was observed primarily in the FP strain resulting in complete agammaglobulinemia within 4 to 6 weeks after transfer of BX donor spleen cells. Serial transfer of this phenomenon was demonstrated in the SC strain.
Activation of Alternate Pathway of Complement by Rabbit F(ab′)2 Fragment
Authors: Fujita T. , Nagaki K. , Inai S.
Published: December 2022

Immune precipitate formed with egg albumin and F(ab′)2 fragment of rabbit anti-egg albumin (F) activated the alternate pathway in normal human serum. F inactivated C3 and C5 without effecting on the early acting components in serum.

On incubation with normal human serum, F combined with several factors (X) to form an intermediate complex, designated FX. FX had an ability to inactivate C3 and C5. The time at which FX showed maximal activity to cleave C3 and C5 (Tmax) was 10 to 20 min at 37°C. Tmax depended on the amount of F added to serum, and increased amount of F shortened Tmax. FX seemed to be composed of at least three factors, factor B, properdin, and C3, judging from the facts that anti-factor B, anti-properdin, or anti-C3 inhibited C3- and C5-cleaving activity of FX. When preformed FX was incubated at 37°C, it decayed to an inactive form, FXd, at 120 min.
A Spin Probe Study of Complement Induced Changes in Membrane Lipids
Authors: Giavedoni E. B. , Mason R. P. , Dalmasso A. P.
Published: December 2022

We reported previously that complement lysis of antibody-sensitized sheep erythrocytes (EA) inhibits the dissociability of membrane (M) lipids by 2.4 M KSCN, in contrast to M lipids from EA lysed hypotonically (Fed. Proc., 34:965, 1975). We interpreted this finding as indicating a complement induced perturbation of M lipids. We now have applied electron spin resonance spectroscopy to investigate changes in the lipid fluidity of M from complement-lysed EA. The spectrum of 5-nitroxide methyl stearate after incorporation into sheep erythrocyte M is characterized by two outer hyperfine extrema. The following parameters, which are sensitive to the motional freedom of the spin label, were measured: The separation of the low-field from the high-field hyperfine extrema (2Az′), the half width at half height of the low-field extrema (Δ1) and of the high-field extrema (Δh). Each of these parameters were similar in M from hypotonically lysed E, EA, and EA treated with C6 deficient rabbit serum.
Complement-Induced Ultrastructural Membrane Lesions: Requirement for Late-Acting Components
Authors: Packman C. H. , Rosenfeld S. I. , Weed R. I. , Leddy J. P.
Published: December 2022

Complement-induced ultrastructural ring-shaped changes in EA have been known since 1963. Initially it was thought that the entire C sequence was necessary for this phenomenon. However, Polley et al., using purified human C components added stepwise to EA, found ring structures on the membrane after the EAC14235 step. Stepwise addition of C6–C9 did not produce more rings, but lysis required the full C sequence. Polley also noted rings on EA exposed to C6-deficient (C6D) rabbit serum, but not C5D mouse serum. Other workers have been unable to find rings on either sheep E or liposomes by using C6D rabbit serum as C source. We have re-examined this problem by using several human sera each genetically deficient in one of the late components.

Sheep EA (108/ml) were incubated for 1 hr at 37°C with an equal volume of a 1:2 dilution of each C-deficient serum (C5D, C6D, C7D, C8D), normal human serum (NHS) and serum heated at 56°C for 30 min (CΔ).
Structural and Genetic Studies on Chicken 7S Immunoglobulin Allotypes
Authors: Wakeland Edward K. , Benedict Albert A. , Abplanalp Hans A.
Published: December 2022

We have previously reported that chicken 7S immunoglobulin (Ig) heavy (H) chain allotypes (CS-1 locus) segregate as phenogroups in F2 progeny. Specificity CS-1.1 formed a phenogroup with CS-1.4 in inbred chicken line UCD 2, and a second phenogroup with CS-1.3 in line UCD 3. To determine whether these phenogroups were formed by combinations of specificities on the same, or on separate subclasses of 7S Ig, their distribution on the 7S Ig molecules of birds homozygous for 7S Ig allotypes was analyzed by radioimmunoassay. Anti-CS-1.1 and anti-CS-1.3 alloantisera each bound more than 94% of line UCD 3 125I-7S Ig. Similar results were obtained with alloantisera to CS-1.1 and CS-1.4 with 125I-7S Ig from line UCD 2. These results indicate that both phenogroups were formed by combinations of specificities present on the same H chain. Thus, each phenogroup described, probably, is the product of a single structural gene which is responsible for more than 94% of the 7S Ig H chain constant regions.

In F1 hybrids with the genotype CS-1.1,1.3/CS-1.2, two populations of serum 7S Ig molecules were detected by direct and sequential binding analysis with specific alloantisera. One population of 7S Ig contained specificities CS-1.1 and CS-1.3, but not CS-1.2; while the second population carried only specificity CS-1.2. Therefore, each population was exclusively the product of one parental allele. Consistent with a genetic regulatory mechanism involving allelic exclusion, no 7S Ig containing allotypes produced by both alleles was detected.
α-Fetoprotein and Human Lymphocyte Subpopulations
Authors: Gupta Sudhir , Good Robert A.
Published: December 2022

Peripheral blood lymphocytes were incubated with varying concentrations of α-fetoprotein and enumerated for total and “active” T lymphocytes, B lymphocytes, and their proliferative responses to phytohemagglutinin. No significant effect was observed on total T or B lymphocyte proportions. However, there was a dose-related increase in proportions of the so called “active” T lymphocytes. the response of human lymphocytes to phytohemagglutinin was markedly depressed. The alteration in the proportion of active T cells and the inhibition of T lymphocyte response to phytohemagglutinin by α-fetoprotein occurred at higher concentrations than are present in amniotic fluid, serum of pregnant women, or serum of adults, but well within the range reached in fetal serum. The immunoregulatory role of α-fetoprotein is discussed.