bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of Properdin in the Alternate Pathway of C Activation
Authors: Götze Otto , Müller-Eberhard Hans J.
Published: December 2022

Properdin (P) was isolated from the euglobulins of human serum. Isolation was accomplished without the use of zymosan by a series of column chromatographic steps, including passage over CM-Sephadex C-50, DEAE-cellulose, Sepharose 6B and CM-cellulose. The final product was homogeneous by Ouchterlony analysis, immunoelectrophoresis, and polyacrylamide gel electrophoresis. Upon immunoelectrophoresis in 1% agarose at pH 8.6, P migrated toward the cathode similar to the behavior of P in aged serum (McLean and Michael, Proc. Soc. Exp. Biol. Med., 1972). The isoelectric point of P was above 9.5, the S-rate, determined by sucrose density gradient ultracentrifugation, was 5.5S. These properties are comparable to those reported by Pensky et al. (J. Immunol., 1968). There was no detectable change in the electrophoretic mobility of isolated C3PA or isolated C3 when either of these components was incubated with P.
Formation, Characterization and Function of a Stable Fluid Phase C5-C9 Complex
Authors: Kolb William P. , Müller-Eberhard Hans J.
Published: December 2022

Previous studies in this laboratory have allowed the formulation of a model for the molecular arrangement of C5, C6, C7, C8, and C9 on the surface of cells undergoing immune cytolysis with an assigned cumulative m.w. of 995,000. To verify directly the existence of a C5–C9 complex, serum samples containing radiolabeled terminal components were activated at 37°C with EA, antigen-antibody complexes, CVF, inulin or zymosan. Subsequent sucrose density gradient ultracentrifugation showed that all treatments cited led to the formation, in varying degrees, of rapidly sedimenting material which incorporated C5, C6, C7, C8, and C9, but not C3. The reaction was inhibited by 0.01 M EDTA and 0°C. The complex had a sedimentation coefficient of 22.4S, a diffusion coefficient of 1.98 × 10-7 cm2 sec-1 and thus a calculated m.w. of 1.04 × 106.
Further Studies on the Effect of Zinc Ions on Guinea Pig C
Authors: Amiraian Kenneth
Published: December 2022

The C titer of guinea pig serum, when treated twice with 0.3 volume of Chelex-100 (Dowex A-1), was reduced from 146 CH50/ml to 12 CH50/ml. Upon addition of 0.00015 M Ca++ and 0.0005 M Mg++, the treated serum had 121 CH50/ml. When adjusted to 6 × 10-6 M Zn++, in the presence of optimum Ca++ and Mg++, the titer was 143 CH50/ml. The optimum zinc ion concentration is dependent upon the concentration of sensitized cells (5 × 10-6 M Zn++ for 5 × 108 cells/7.5 ml). The effect of Zn++ is unrelated to the enhancement of guinea pig C by the zinc metalloenzyme carboxypeptidase A. Cadmium and nickel ions are also effective in the presence of optimum CA++ and Mg++. Cobalt has only a slight effect while manganese has no effect on guinea pig C. Kinetic studies indicate that zinc ions exert an inhibitory effect early in the immune hemolytic reaction and that enhancement is seen only after 16 min at 36°C.
Mechanism of Activation of C1 by Trypsin
Authors: Loos M. , Borsos T.
Published: December 2022

Trypsin activates C1 in serum as well as C1 bound to EAC4. At 37°C about 500 µg trypsin/ml are necessary for optimal activation of C1 in whole serum; 1 µg trypsin/ml is necessary to activate C1 in diluted serum (1:3000) and C1 bound to EAC4. The activation of precursor C1 (EAC14) to C1̄ (EAC1̄4) with or without trypsin is a time and temperature dependent reaction. The activation energy of the reaction EAC14 to EAC1̄4 is 7.5 kcal/mole and in the presence of trypsin 7.6 kcal/mole; the temperature coefficients (Q10) of both reactions are 1.57 and 1.55, respectively. Although there was no difference in the activation energies of both reactions, the reaction velocities were different. At a given temperature the value of the reaction velocity of the EAC1̄4 generation in the presence of trypsin was twice as high as in the reaction without trypsin.
Molecular Restriction of Anti-DNP Antibodies Induced by Dinitrophenylated Type III Pneumococcus
Authors: Montgomery Paul C. , Pincus Jack H.
Published: December 2022

Rabbits were hyperimmunized with ε-DNP-lysine covalently coupled to type III encapsulated pneumococci (DNP-Pn vaccine). After a preimmunization with pneumococcal vaccine followed by immunization with DNP-Pn vaccine, 11 of 13 rabbits produced anti-DNP antibodies with titers ranging from 0.28 to 1.30 mg/ml. The affinities of the anti-DNP antibodies were low (105 liters/mole). The molecular heterogeneity of 7 of the 11 responding rabbits, as determined by isoelectric focusing, was markedly reduced. Upon continued immunization some rabbits maintained or increased their restriction while others exhibited a loss of restriction and evidence of molecular heterogeneity. Although titers and heterogeneity generally increased during prolonged immunization, association constants for the anti-DNP antibodies remained in the range of 105 liters/mole. The present experiments indicate that a bacterial carrier can have a marked effect on the structural complexity of anti-DNP antibodies and provide a new means of obtaining structurally restricted anti-hapten antibodies.
Alternative Potentiating and Inhibitory Effects of GVH Reaction on Formation of Antibodies Against a Thymus-Independent Polysaccharide (SIII)
Authors: Byfield Patricia , Christie George H. , Howard James G.
Published: December 2022

The direct PFC response to SIII (type 3 pneumococcal polysaccharide) has been studied in adult (C57BL × CBA-T6T6) F1 mice in which graft-vs-host (GVH) reaction was induced by parental strain lymphoid cells. The response was potentiated for 3 to 6 days after injection of lymphoid cells when SIII was injected either separately or coupled to the cells. An analysis of this effect revealed: a) that it requires recognition of the host by viable donor thymus-derived (T) lymphocytes and their subsequent proliferation (the reverse, host-vs-graft, situation was ineffective); and b) that it seems to be attributable to a different T cell function than that responsible for specific “helper” activity.

On the other hand, F1 mice examined 6 to 109 days after injection of 8 × 107 parental spleen cells were totally unresponsive to SIII. Furthermore, there was no response to SIII in lethally irradiated mice repopulated with spleen cells from GVH animals or with a mixture of GVH and normal spleen cells. Treatment of the GVH cells with anti-ϑ serum partially alleviated this suppressive effect.

We conclude that potentiation and suppression are both attributable to T cell activity, and that they seem to differ qualitatively rather than quantitatively. Macrophages may be involved in an intermediary effector capacity in the suppressive phenomenon.
Genetics of the Immune Response
Authors: Pauli Raimund Di
Published: December 2022

Cross-reactivity of IgM antibodies, against a lipopolysaccharide, has been used as a marker for the variable part of these antibodies. BALB/c and DBA/2An differ in the antibody specificity produced against the same lipopolysaccharide. Backcross data are compatible with a control by a single gene. About half of the F1 animals resembled the BALB/c parent and the other half the DBA/2An parent. This suggests that only one allele is expressed in heterozygous animals.
Presidential Address to the American Association of Immunologists, Delivered in Atlantic City, New Jersey, April 10, 1974
Authors: Benacerraf Baruj
Published: December 2022

Fellow immunologists, I would like to express my deep appreciation for the great honor you have bestowed upon me by electing me President of our Association. I particularly value the privilege of addressing you today, as it has been the custom for previous Presidents of the American Association of Immunologists. Presidential addresses constitute a considerable challenge, since traditionally neither the subject matter nor the form has been defined either by our bylaws or by past custom. I have debated with myself for several weeks what would be a matter worthy of your attention today. A survey of previous presidential addresses revealed that former Presidents of our Association have taken this opportunity most frequently to discuss some phase of their work which best exemplified their scientific contributions to immunology. Many others have chosen to make a historical survey of the development of the field of immunology or of the progress of our Society.
Effects of Interferon on Antibody Synthesis in Vitro
Authors: Gisler Roland H. , Lindahl Pernilla , Gresser Ion
Published: December 2022

Mouse interferon preparations inhibited the antibody response to sheep erythrocytes (SRBC) in spleen cell cultures. The effect was seen when the cultures were pretreated with interferon for 6 hr, or when interferon was added up to 40 hr after addition of SRBC. The factor responsible for this inhibition could not be dissociated from the antiviral activity of interferon by standard physicochemical means. The application of a mosaic cell culture system suggested that the inhibitory effect of interferon was due to an action on B cells. Interferon treatment of T cells and macrophages did not affect their normal function as helper cells in this system. To our knowledge, these findings constitute the first example of an effect of interferon on B lymphocytes.

In addition, it was found that in “low-responder” composite cultures, pretreatment of the B cell population with low amounts of interferon led to a considerable enhancement of the antibody response, whereas higher concentrations of interferon gave a similar degree of inhibition as that found for “high-responder” cultures.

Late addition of mouse interferon preparations to SRBC-stimulated spleen-cell cultures augmented the number of plaque-forming cells (PFC). This effect was most pronounced when interferon was added only 4 hr before the plaques were counted on day 4. This increase was followed on day 5 by a decrease in the number of PFC compared with that of control cultures.
Immunologic Reactions to Haptens on Autologous Carriers
Authors: Walters Curla S. , Moorhead John W. , Claman Henry N.
Published: December 2022

Antigen-induced lymphocyte proliferation in vitro was used to investigate changes in receptor affinity of cells from mice immunized with haptens coupled to autologous carriers. We attempted: 1) to show a differential sensitivity to antigen in vitro after priming with low or high doses of antigen in vivo; 2) to assess the in vitro response to different doses of antigen at various times after priming with a single dose of antigen; 3) to demonstrate high affinity cells by blocking the in vitro response with low doses of free hapten. Using all three procedures, we were unable to demonstrate either a selection of high affinity cells or a maturation of cells with high affinity receptors with time after immunization. The results suggest that in mice, hapten coupled to autologous carriers are weak immunogens and are unable to prime a majority of cells with high affinity receptors.
Effect of Carrier Priming on the Distribution of Anti-Hapten Antibodies between IgG1 and IgG2 Isotypes in the Hyperimmune Guinea Pig
Authors: Liu Sammy H. , Koo Peter H. , Cebra John J.
Published: December 2022

Priming of guinea pigs with a soluble carrier protein, keyhole limpet hemocyanin, can have a marked influence on the isotype distribution between IgG1 and IgG2 of anti-dinitrophenyl or anti-p-azobenzenearsonate antibodies produced upon subsequent immunization with hapten-carrier conjugates. Lowdose carrier priming (40 µg) causes a marked shift from IgG2 to IgG1 anti-hapten antibody without affecting the total amount of antibody produced compared with the anti-hapten response of animals directly immunized with conjugates. High-dose carrier priming (300 µg) enhances the overall anti-hapten response by causing an increase in the amount of IgG2 anti-hapten antibody produced while an equivalent amount of IgG1 antibody to that made after low dose carrier priming is also synthesized. Preliminary studies indicate that injection of a low dose of soluble carrier depresses MIF release by carrier-reactive T lymphocytes. We postulate that such prior injection of carrier also affects the balance between the “helper function” reactivity of carrier-specific T cells and the sensitivity to stimulation of different isotope-committed B cells.
Recurrent Experimental Allergic Encephalomyelitis in the Lewis Rat
Authors: McFarlin Dale E. , Blank Susan E. , Kibler Robert F.
Published: December 2022

Experimental allergic encephalomyelitis (EAE)3 is an immunologically mediated disease produced in genetically susceptible animals by the injection of central nervous system (CNS) tissue or myelin basic protein emulsified in complete Freund's adjuvant. Because of the demyelinating character of the CNS lesions, EAE has served as a useful working model for the human demyelinating diseases, in particular multiple sclerosis. One limitation of the experimental model has been that, unlike multiple sclerosis, it is primarily a monophasic illness. Although recurrences have been induced in EAE (1, 2), spontaneous relapses have been infrequent and unpredictable (3). During studies of the factors involved in the immunoregulatory mechanisms of EAE in the Lewis Rat (4) we observed that, by providing supportive care during the acute episode, the majority of rats rapidly recovered and approximately 1 week later developed a second clinical episode, albeit milder than the first. The frequency of the second episode was age related.
Studies on the Human T-Lymphocyte Population
Authors: Owen Frances L. , Fanger Michael W.
Published: December 2022

The IgG fraction of a goat antibody prepared against human infant thymus cells and extensively absorbed with pooled human red blood cells, insolubilized human Ig and cells from a patient with chronic lymphocytic leukemia (ATG), was used to study human peripheral lymphocyte populations. ATG inhibited the response of human peripheral lymphocytes to concanavalin A (Con A) but not to pokeweed mitogen (PWM). The F(ab′)2 and Fab′ fragments of ATG were as effective as ATG in inhibiting Con A induced 3H-thymidine incorporation. ATG did not interact with Con A or the receptor for Con A on the lymphocyte but was inhibitory in the absence of complement as a result of its apparent selective cytotoxicity. The antibody also inhibited 3H-thymidine incorporation in the one-way mixed lymphocyte reaction, again without affecting the response of these cells to PWM. The specificity of ATG for the T-lymphocyte population was also suggested by fluorescent labeling experiments which indicated that ATG reacted with only 50 to 80% of human peripheral lymphocytes. These results suggest that ATG is directed toward antigens unique to the human T-lymphocyte population. Moreover, it appears that ATG can be used to study human T-lymphocyte responses and to follow the isolation of T-specific antigens.
Similarities of Fc Receptors in Human Malignant Tissue and Normal Lymphoid Tissue
Authors: Tønder Olav , Morse Paul A. , Humphrey Loren J.
Published: December 2022

The Fc receptor activity in various tissues has been studied using hemadsorption to cryostat sections of fresh frozen tissue or rosette formation with single cells in suspension. Indicator cells (EA) were sheep erythrocytes sensitized with various amounts of rabbit IgG antibodies (A). Malignant tissues and cells showed very similar behavior to lymphoid tissue and peripheral blood lymphocytes and monocytes, although adsorption of non-complexed A to monocytic areas of spleen was quantitatively stronger than to B lymphocytic areas and malignant tissues. Receptor activity measured as hemadsorption and the percentage of rosetting cells varied with the degree of sensitization of EA with lymphocytes, e.g., from 58% rosettes using EA sensitized with 1 agglutinating unit of A, to less than 1% with EA 1/16 unit. This indicates that Fc receptor is not a property of distinct subpopulations of the reactive cell types. Apparently, the presence and availability of this receptor fluctuate in or on each cell. Treatment of tissue sections or single cells with neuraminidase in various concentrations showed a similar enhancing effect on malignant cells, lymphocytes, and monocytes. Apparently, the Fc receptors possessed by the three cell types are similar in nature and properties.
Clonal Heterogeneity of the Anti-Erythrocyte Autoantibody Responses of NZB Mice
Authors: De Heer David H. , Edgington Thomas S.
Published: December 2022

The pathogenetic anti-erythrocyte autoantibody responses of NZB mice were characterized at the level of the autoantibody secreting B lymphocyte. Clonal heterogeneity was evaluated by reference to certain phenotypic expressions of immunoglobulin heavy chain constant and variable region genes. The anti-X autoantibody response appeared completely restricted to the NZB strain and was predominantly of IgG class. Specific anti-X B2 lymphocytes exhibited marked clonal diversity in regard to immunoglobulin class and heterogeneity of autoantibody affinity as determined by competitive plaque inhibition assays. The anti-HB autoantibody response, a genetically non-restricted response, was predominantly of IgM class; splenic anti-HB B2 lymphocytes also exhibited marked clonal diversity in respect to immunoglobulin class and binding affinity of the secreted autoantibody. Neither response exhibited a maturational increase in homogeneity similar to that observed in anti-sheep red cell responses by NZB mice. These data indicate that heterogeneous sets of B lymphocytes are recruited in these responses, an observation consistent with the concept that the immunogenesis of these pathogenetic autoimmune responses may be immunoregulatory in nature.
A Fragment from the Constant Portion of IgG Obtained by Peptic Digestion at High Temperatures
Authors: Seon B.-K. , Pressman D.
Published: December 2022

Two myeloma IgG immunoglobulins and normal human IgG immunoglobulin were digested with pepsin at various temperatures for various times. After digestion at temperatures exceeding 60°C, all the proteins studied showed a distinctive anodic arc which had not been observed after digestion at 37°C. An immunoglobulin fragment corresponding to this arc was isolated from the peptic digests of the myeloma immunoglobulins and the normal immunoglobulin by gel filtration on a Sephadex G-150 column. The amino acid compositions of these high temperature fragments from the three immunoglobulins were as consistent as the compositions of the constant portions of different IgG molecules.

Sodium dodecyl sulfate (SDS) acrylamide gel electrophoresis of the reduced fragment of IgG (Col) showed only two bands. That of normal human IgG showed two strong bands at the identical positions to the two bands of IgG (Col), in addition to a weak third band. The molecular weights of these two bands correspond to the approximate size of a domain, i.e., 12,000 and 14,000. An immunodiffusion experiment indicated that the high temperature fragment was derived from the constant portions of light and heavy chain. This fragment sedimented as a single symmetrical peak with a s20,w value of 3.3 in pH 8.6 Veronal buffer (ionic strength 0.10). It appears to be a new fragment which has not yet been reported.
Transformation of Human Peripheral Lymphocytes by Galactose Oxidase
Authors: Dixon James F. P. , Parker John W. , O'Brien Richard L.
Published: December 2022

Human peripheral lymphocytes can be transformed by treatment with galactose oxidase alone. Prior treatment with neuraminidase enhances this effect. The aldehyde blocking agents thiocarbohydrazide, hydroxylamine, dimedone, and sodium borohydride block transformation when they follow, but not when they precede, galactose oxidase treatment. Thus, as is the case for periodate-induced lymphocyte transformation, the formation of free aldehyde at the cell surface would seem to be a critical event in the triggering of transformation by this agent.

The degree of transformation is highly variable from individual to individual, and also for the same donor at different times. However, the lymphocytes of some people give a consistently poor response to galactose oxidase. Similar results have been obtained for periodate-induced transformation of human lymphocytes, but to date this is unexplained.
Listeria Cell Wall Fraction: A B Cell Adjuvant
Authors: Campbell Priscilla A. , Schuffler Carol , Rodriguez Gilberto E.
Published: December 2022

A crude cell wall-rich fraction of Listeria monocytogenes (LCWF) has previously been shown to induce resistance to Listeria infection in mice, to be a murine B cell mitogen, and to be an immunologic adjuvant. Data reported here show that LCWF stimulates immune responses by three different murine B cell populations that respond poorly or not at all to thymus-dependent antigens when T cells are severely depleted or absent. These B cell populations include nude mouse spleen cells, anti-θ serum-treated spleen cells and bone marrow-derived spleen cells. In addition, sheep erythrocytes coated with LCWF are converted to thymus-independent antigens. The adjuvant effect of LCWF on B cell populations is not enhanced by the presence of T cells, nor does LCWF increase in vivo education of T cells to sheep erythrocyte antigens. Thus, a direct effect of LCWF on T cells could not be demonstrated, suggesting that LCWF does not act on T cells. Because of these LCWF effects on B cells, the possible role of B cells in resistance to infection by Listeria monocytogenes is discussed.
Inhibition of Mitogen-Induced Lymphocyte Transformation by Local Anesthetics
Authors: Ferguson Ronald M. , Schmidtke Jon R. , Simmons Richard L.
Published: December 2022

Chlorpromazine (CPZ) and lidocaine were added to cultures of mouse spleen cells stimulated by concanavalin A (Con A), phytohemagglutinin (PHA), pokeweed mitogen (PWM) and lipopolysaccharide (LPS). Concentrations of CPZ greater than 5 × 10-6M and concentrations of lidocaine greater than 2 × 10-3M totally inhibited the mitogenic responses to all four mitogens. Minimal inhibitory concentrations of neither drug interferred with cell viability as determined by trypan blue uptake or 51Cr release. The effects were totally reversed by the removal of the drugs from the culture. Addition of the drug at intervals after mitogen exposure demonstrated that the inhibited event occurred relatively soon after exposure to the mitogen. For example, the addition of lidocaine or CPZ more than 24 hr after Con A stimulation had no effect on tritiated thymidine incorporation. Elevated concentrations of cyclic AMP, cyclic GMP (or their derivatives) or calcium ions could not reverse the local anesthetics inhibition. The known membrane active actions of these drugs and the rapid reversibility of the effect strongly support the idea that the local anesthetics act on the surface membrane of lymphocytes. Binding of radiolabeled Con A or LPS to lymphocyte membranes in the presence of lidocaine or CPZ was not inhibited. The possibility exists that CPZ and lidocaine disorganize cell membranes so as to interfere with the surface membrane elaboration or action of a second messemger, or interfere with cell-cell interactions.
Lysis of Tumor Cells by Antibody and Complement
Authors: Boyle Michael D. P. , Hoanian Sarkis H. , Borsos Tibor
Published: December 2022

The ascites form of a chemically induced guinea pig hepatoma, line-10, was resistant to killing in vitro by xenogeneic antibody and guinea pig complement. Pretreatment of line-10 cells with certain proteolytic enzymes rendered them susceptible to the killing action of antibody and guinea pig complement. The effects of enzyme pretreatment were dependent on enzyme concentration, temperature, and could be blocked by addition of competitive or non-competitive inhibitors. The effect of the enzyme treatment could be reversed by incubating the treated cells at 37°C (but not at 0°C), in the absence of the enzyme. Effective enzymes included ficin, bromelain, pronase, elastase, papain, trypsin, collagenase, lipases type I and type VI, and the neuraminidase preparation isolated from Clostridium perfringens. The activity of the lipase preparations and the neuraminidase preparation isolated from Clostridium perfringens appeared to be caused by proteolytic enzyme contamination. Enzyme preparations that proved ineffective in rendering the line-10 cells sensitive to killing by antibody and guinea pig complement included DNase, RNase, β-glucuronidase type 6A or type B10, hyaluronidase type V or type VI, and pectinesterase.

Enzyme pretreatments that rendered the cells sensitive to killing by antibody and guinea pig complement also enhanced the sensitivity of the cells to the killing action and human complement. In addition the neuraminidase enzyme preparations from Vibrio cholorea or influenza virus increased the sensitivity of the line-10 cells to killing by antibody and human complement, but not to killing by antibody and guinea pig complement.
Humoral Immunity in Aged Mice
Authors: Segre Mariangela , Segre Diego
Published: December 2022

The secondary response of young-adult and old BC3F1 mice to DNP was assessed by boosting spleen cells from primed mice with DNP-KLH, propagating them in diffusion chambers implanted into irradiated recipient mice, and by counting the DNP-specific PFC. There was no difference between young-adult and old cells in the antigen dose response and in the kinetics of the immune response. There were marked differences in the magnitude of the peak response. The response of 2-year-old mice was significantly smaller than the response of young-adult mice. Two and one half-year-old mice had the smallest response. In the old mice, there was an inverse correlation between the number of nucleated cells in the spleen and the log of the number of PFC.
Development of Rat Mast Cells in Vitro
Authors: Ishizaka Teruko , Okudaira Hirokazu , Mauser Linda E. , Ishizaka Kimishige
Published: December 2022

Mast cells were differentiated by long-term culture of rat thymus cells on rat embryonic fibroblast monolayers. Mature mast cells obtained in the culture were morphologically similar to normal peritoneal and thoracic mast cells and possessed specific receptors for IgE on their surface. In culture, blast cells appeared on the monolayer several days after seeding of thymus cells. These cells developed into young mast cells in the monolayer and became free in the culture medium with maturation. Receptors for IgE were detected on the surface of mastoblasts which contained a small amount of metachromatic granules. Evidence was obtained which suggested that the number and/or affinity of the receptors for IgE increases with maturation of mast cells. It was found that some mast cells differentiated from monolayers of embryo cells without seeding thymus cells. The present experiments, however, clearly showed that mast cells can be differentiated from thymus cell culture without monolayer. It appears that both thymus and embryo tissues contain precursors of mast cells.
Serum Factors Activating the Alternative Complement Pathway in Autoimmune Disease: Description of Two Different Factors from Patients with Systemic Lupus Erythematosus
Authors: Arroyave Carlos M. , Wilson Merlin R. , Tan Eng M.
Published: December 2022

Serum factors which activated the alternative pathway of complement were detected in 10 of 26 patients with systemic lupus erythematosus (SLE), three of 18 patients with mixed connective tissue disease, one patient with scleroderma, and one with Sjögren's syndrome. This activation was detected by conversion of factor B and C3 into split products and by lysis of glutathione-sensitized human erythrocytes under conditions where classical pathway activation was blocked. The serum factors capable of activating the alternative pathway could be separated into 7S and 19S-type molecules by sucrose density gradient ultracentrifugation. In two patients with SLE, serum factors were isolated by ion-exchange chromatography, preparative electrophoresis, and molecular sieve chromatography. The serum 7S factor was immunochemically identical to the C3 nephritic factor (C3NeF) of hypocomplementemic chronic glomerulonephritis. Antisera to other complement components failed to react with this purified material. The 7S factor was able to activate the alternative pathway in C2 deficient serum, and in normal human serum under conditions where classical pathway was blocked, but was unable to do so in sera depleted of factor B, factor D, and C3. The activity could be removed by antiserum to C3NeF. In contrast, the serum 19S activator of the alternative pathway was not reactive with antiserum to C3NeF. It had a fast gamma mobility on electrophoresis and the activity could be removed by absorption with anti-IgG and anti-IgM. It was suggested that the 19S factors could consist, in part, of immune complexes.
Radiolabeling of Immunoglobulin without Loss of Antibody Activity—Use of 14C-Phenylisothiocyanate with Human Cytotoxic Antibody
Authors: Levy Nelson L. , Dawson Jeffrey R.
Published: December 2022

Radioiodination of immunoglobulins, particularly human cytotoxic IgG, induced a marked loss of antibody activity. Phenylisothiocyanate (PITC) readily reacts with α-amino groups and the ε-amino groups of lysines to form phenylthiocarbamyl derivatives. Since PITC is commercially available with 14C, 3H, and 35S substitutions, it provided an alternative means for labeling immunoglobulin which preserved antibody activity. There were approximately 80 PITC binding sites per IgG molecule, and 14C-PITC was bound with an efficiency in excess of 80%. With as many as 40 PITC molecules bound per IgG molecule, full cytotoxic activity was retained by both anti-HLA isoantisera and human anti-melanoma autoantisera. Even with 70 to 80 molecules of PITC bound per IgG molecule, over 80% of the antibody activity remained. Radiolabeling of antibody with 3H, 14C, or 35S-substituted PITC will greatly facilitate studies of antibody binding, particularly to cell surface antigens.
Differentiation of Lymphoid Cells: B Cell As a Direct Target and T Cell As a Regulator in Lipopolysaccharide-Enhanced Induction of Immunoglobulin Production
Authors: Shinohara Nobukata , Kern Milton
Published: December 2022

The cells involved in the stimulatory effect of bacterial lipopolysaccharide (LPS) on the induction of immunoglobulin (Ig) production by rabbit spleen cells cultured in the absence of antigen has been analyzed. Addition of LPS caused a several-fold enhancement of both DNA synthesis and Ig production. These enhanced activities were not significantly affected by depletion of adherent cells in the spleen cell population. Although inactivation of splenic T cells by anti-thymocyte serum (ATS) treatment did not affect the enhancement of DNA synthesis due to LPS, such treatment did adversely affect the enhancement of Ig production by LPS. Furthermore, the enhancement of Ig production of ATS-treated spleen cells by LPS was found to be dependent on the number of thymocytes added. In addition, the prior incubation of ATS-treated spleen cells with LPS resulted in effective enhancement of Ig production when such ATS-treated spleen cells and thymocytes were combined after removal of LPS. An identical experiment, except that thymocytes instead of ATS-treated spleen cells received the prior incubation with LPS, did not result in enhancement of Ig production. Finally, the enhanced Ig production due to LPS was inhibited by hydroxyurea, a known inhibitor of cellular DNA synthesis. The relationship between the mitogenic activity of LPS on B cells, the regulatory function of T cells, and the enhancement of Ig production by LPS is discussed in relation to the contrasting reports concerning the cellular target of LPS.
Specific Suppression of the Immune Response by Hgg Tolerant Spleen Cells
Authors: Doyle Michael V. , Parks D. Elliot , Weigle William O.
Published: December 2022

After adoptive transfer, the spleen cells from mice made tolerant to human γ-globulin (HGG) specifically suppress the immune response of normal spleen cells. However, this suppressive activity in the spleen cells of tolerant mice is only present for a brief period after treatment with tolerogen. Spleen cells from animals injected 10 days earlier with tolerogen reduce the immune response of an equal number of normal spleen cells by 75%. Spleen cells from mice made tolerant 40 days previously are, however, no longer suppressive, even though they remain completely unresponsive. These data suggest that active suppression of antigenreactive cells is not the mechanism responsible for maintaining tolerance to HGG, but rather is only transiently associated with the tolerant state. Further evidence in favor of the separation of the tolerant state from suppressive activity is that complete suppression of the normal spleen cell response requires either a high ratio of tolerant to immune competent cells or a delay in the antigenic challenge of the reconstituted recipients. By contrast, such manipulations are not required to demonstrate the complete unresponsiveness of the tolerant cells after adoptive transfer.
Influence of Colony Type on Susceptibility of Gonococci to Killing by Human Serum
Authors: McCutchan J. Allen , Levine Sharon , Braude Abraham I.
Published: December 2022

The virulence of gonococci is related to colony morphology and has been attributed to pili found only on the virulent types. Because complement-mediated serum killing is a major defense against other Gram-negative bacteria, we tested the resistance to normal human serum of virulent and avirulent types from two strains. Virulent types were more resistant than avirulent types. This stable, relative serum resistance of virulent colony types persisted during prolonged passage on agar. In addition, virulent types acquired complete resistance after exposure to serum which was rapidly lost during passage on agar. This acquired, complete resistance helps explain the observation that freshly isolated gonococci are resistant to their host's serum, but lose this resistance when passed on agar. The capacity to become resistant to the serum of newly infected patients may contribute to virulence of T1 and T2 gonococci.
Evidences for a “New” C1 Inhibitor, Factor S, Released from Leucocytes
Authors: Bernard A. , Boumsell L. , Good R. A. , Borsos T. , Day N. K.
Published: December 2022

Pretreatment of sheep erythrocytes (E) with supernatants of short-term cultures of mouse spleen or thymus cells induces the following interactions with guinea pig complement: 1) Supernatant pretreated EAlim (S.EAlim) are less susceptible to lysis by excess complement. S.EAC1̄4 yield a lower C2 titer, have an increased Tmax, but an unchanged C2 decay when compared with medium-pretreated EAC1̄4. Lysis of already formed EAC1̄4̄2̄is not inhibited. Therefore the inhibition affects C2 activation. 2) E, pretreated with supernatant (S.E), are able to fix C1̄, as detected by transfer to EAC4: In order to generate the same number of C1̄-fixing sites by using IgM hemolysins, five to six times the optimal amount is required. Yet S.EC1̄ cannot be lysed by the other components added in excess. S.EC1̄ can remove C4 from the fluid phase with the same efficiency as EIgMC1̄ but S.EC1̄ treated with C4 cannot remove C2. 3) Native C1̄ binds to S.E: S.E and EIgM remove C1 from the fluid phase with the same efficiency; fixation of 1̄ on S.E is decreased after incubation with native C1̄
Complement Lesions in Cell Membranes from Joint Effusions of Various Types of Arthritis
Authors: Dourmashkin R. R. , Patterson S.
Published: December 2022

Electron microscopy of differentially centrifuged synovial fluid from patients with differing types of arthritis revealed a few membranes bearing complement lesions. This finding occurred equally in patients with seronegative arthritis as well as in patients with rheumatoid arthritis and positive rheumatoid factor. By incubating effusion leukocytes in vitro with effusion fluid, and peripheral leukocytes with syngeneic serum complement, under conditions in which classical complement activation was inhibited, it could be shown that this finding was through activation of the alternative pathway, by means of lysosomal enzymes. It is not clear where, in inflammatory exudates, is the site of action; it may be on the internal membranes of already damaged cells, as no detectable cytotoxic effect could be found by using 86Rb as a marker for specific release in PMN leukocytes. By means of hemolysis of indicator cells, lysosomal enzyme was found to have the net effect of inhibiting the lysis of indicator cells in the presence of complement activated by the alternative pathway.
Stimulation of Murine B Lymphocytes by Isolated C3b
Authors: Hartmann K.-U. , Bokisch V. A.
Published: December 2022

Purified human C3b added to mouse lymphocyte suspensions triggers activation of B lymphocytes. Activation of up to 50% of the cells was detected by increased incorporation of 3H-thymidine, blast cell formation, and antibody secretion (polyclonal development of plaque-forming cells).

Activation was observed in suspensions of lymph node and spleen cells of all mouse strains tested, including DBA/2, BALB/c, C3H/HcJ and Nu/nu, the congenitally thymus deficient strain. Removal of the rapidly adhering or thymusderived cells did not reduce activation. Activation was dependent on the dose of added C3b, which was varied between 1 and 50 µg/0.5 ml culture, and was potentiated by the presence of fetal calf serum. Addition of soluble rabbit anti-C3 reduced the stimulatory activity of C3b, and Sepharose-bound anti-C3 removed the activity from the C3b solution. C3a or C3c did not activate the lymphoid cells, and addition of C3b inactivator (partially purified from human serum) strongly inhibited the stimulation.
Activation of Guinea Pig B Lymphocytes by Fragments of the Third Component of Complement
Authors: Koopman W. J. , Sandberg A. L. , Wahl S. M. , Mergenhagen S. E.
Published: December 2022

Recent evidence suggests that the third component of complement (C3) may play an important role in regulating immune function at the cellular level. Fragments of C3, derived by proteolysis of purified guinea pig C3, were tested for their capacity to induce mediator production and proliferation in guinea pig lymphocytes in vitro. Trypsin digestion of C3 yielded C3a and C3b (1% trypsin, 2 min, 20°C) or C3c and C3d (2% trypsin, 1 hr, 20°C). These fragments were separated either by Sephadex G-75 (C3a and C3b) or Bio-Gel P-100 (C3c and C3d) column chromatography and were characterized both immunoelectrophoretically and functionally by rosette inhibition. Guinea pig B lymphocytes produced a mononuclear chemotactic factor when stimulated with C3b and, to a lesser extent, with native C3, as demonstrated by the presence of chemotactic activity in supernatants assayed in modified Boyden chambers. These fragments did not induce cell proliferation.
Different Behaviour of Native and Purified C1q, a Subunit of the First Component of Complement
Authors: Loos M. , Hill H. U.
Published: December 2022

The first component of complement (C1) consists of three distinct subunits, C1q, C1r, and C1s. Two different forms of C1r and C1s were described, a proesterolytic (native) and esterolytic (active) form. Our experiments suggest that C1q also exists in two different forms. After centrifugation of undiluted human serum on a sucrose gradient (5% to 30%, pH 7.35, µ = 0.06, 16 hr, 4°C, 39,000 rpm) C1 activity was found in the 19S region. After centrifugation of diluted human serum (1:50 in 5% sucrose buffer, pH 7.35, µ = 0.06) no C1 activity was found. The loss of C1 activity was due to the loss of C1q activity, whereas the activity of the subunits C1r and C1s was detected in the 7S and 4S region, respectively. Under similar conditions the hemolytic activity of purified C1q was not affected. Addition of purified C1q to diluted human serum led to a reassociation of purified C1q with C1r and C1s in serum as tested hemolytically as well as by ultracentrifugation.
Techniques of Fractionating Complement in-Vivo by the Use of Activated Complement Components to Trigger the C3b Feedback Cycle
Authors: Mak L. W. , Lachmann P. J. , Majewski J.
Published: December 2022

Cobra venom factor is a highly effective complement-activating agent in vivo by virtue of its ability to trigger the C3b feedback cycle. An investigation has been made to see whether triggering of the C3b feedback cycle by activated autologous complement components could similarly provide a mechanism for in vivo complement activation.

It has been found that C4oxy2 added to serum triggers the C3b feedback cycle, effectively in vitro. Similar C3b, Bb can be used as the triggering enzyme.

Experiments have been performed in vivo in rats showing that significant complement activation accompanied by the leukocyte changes characteristic of CVF treatment can be produced with the altered complement components. Such techniques promise to be applicable to complement (in)activation in vivo in man.
Participation of the Alternative Pathway in Complement Metabolism in Systemic Lupus Erythematosus
Authors: Wilson M. R. , Arroyave C. M. , Tan E. M.
Published: December 2022

In order to determine if in vivo activation of the alternative pathway of complement was present in systemic lupus erythematosus (SLE), highly-purified Factor B was labeled with radioactive iodine (125I), and its metabolism studied in 11 patients with SLE and 6 control subjects. Serum factors, which could activate the alternative pathway of complement in in vitro assays were demonstrated in 6 of 11 patients with SLE. These serum factors were detected by lysis of glutathione-sensitized human erythrocytes and by C3 and Factor B conversion in the presence of EGTA (10 mM) and MgCl2 (0.3 mM) conditions which block activation of the classical pathway but permit activation of the alternative pathway. All 6 patients with serum factors capable of activating the alternative pathway in vitro, had in vivo evidence of alternative pathway activation as measured by an increased fractional catabolic rate (FCR) of Factor B.
Absence of demonstrable effects of thymocytotoxic antibodies on serum thymic hormone level in NZB mice
Authors: Gershwin M. Eric , Steinberg Alfred D.
Published: December 2022

Sir: We are pleased to read the new data provided by Bach, Dardenne, and Bach on the critical question: Is a decrease in thymic hormone(s) levels responsible for the development of autoimmunity in New Zealand mice? There is no question that these workers have provided the world with important ideas and information regarding autoimmunity and thymic factor(s). We believe, however, that Drs. Bach and co-workers have misinterpreted the thrust of our paper. Contrary to their assertion above, we do not conclude that the decrease in “thymic hormone levels” is due to NTA. Rather, we conclude that insufficient information is available to implicate decreased thymic hormone(s) levels as the cause of autoimmune disease in New Zealand mice. Most difficult to explain are the immunologic abnormalities in NZB and especially in NZB/NZW mice before the fall in thymic hormone(s) as measured by the mouse rosette assay.
Immunologic Characteristics in Relation to High and Low Leukemogenic Activity of Radiation Leukemia Virus Variants
Authors: Haran-Ghera Nechama , Rubio Nazario
Published: December 2022

Comparative analysis of immune responses induced by two variants of the radiation leukemia virus having high (A-RadLV) or low (D-RadLV) leukemogenic potential was performed. Effective immunization could be induced only with RadLV variants having low leukemogenic potential when injected into adult mice of specific strains, (D-RadLV in C57BL/6 mice and A-RadLV in (BALB/c × C57BL/6)F1 mice). Immunologic cross-reactivity among the RadLV variants and the leukemic cells induced by them was demonstrated in several systems. Transplantation resistance could be induced in certain strains against challenge of leukemic cells induced by both RadLV variants. Isoantisera, raised in the suitable strains, could neutralize effectively the leukemogenic activity of both A-RadLV and D-RadLV. Leukemic cells induced by each of the variants shared cell-surface antigens. Circulating anti-tumor antibodies, induced by both variants in the appropriate strains, lysed efficiently the target leukemic cells.
Activation of the Classical Complement Pathway by Fc Fragment of Human IgA
Authors: Burritt Mary F. , Calvanico Nickolas J. , Mehta Satish , Tomasi Thomas B.
Published: December 2022

Intact Fcα fragments from five human IgA myeloma proteins were produced by two different methods, high temperature trypsinolysis and IgA protease digestion. Three of the Fcα fragments activated the classical pathway as determined by the titers of the individual complement components. Two other fragments did not fix complement by either pathway.
Physicochemical Characterization of Complement-Interacting Factors from Taenia taeniaeformis
Authors: Hammerberg Bruce , Williams Jeffrey F.
Published: December 2022

Complement-fixing activity was washed from the surface of metacestodes of Taenia taeniaeformis and characterized physicochemically by enzymatic digestion, gel filtration, anion exchange chromatography, ultracentrifugation, and gel electrophoresis. The results provided evidence of an association between antihemolytic activity and macromolecules containing carbohydrate, protein, sulphate, and hexosamine, heterogeneous in net negative charge but resistant to proteolysis and β-elimination reactions, and free of sialic and uronic acids. The most active fraction was eluted from DEAE-cellulose with 0.3 M NaCl in 0.01 M Tris-HCl, pH 8.0, and when parasites were incubated with a source of 35SO4--, radioactivity was incorporated into molecules eluting under identical conditions. Active chromatographic fractions precipitated with protamine sulphate, suggesting that they were polyanionic in nature. There was some evidence of dissociation of the active substance on polyacrylamide gels where fast moving sulphate-containing bands were observed which no longer reacted with protamine or C.

The results are consistent with the possibility that the active substance is a polysulfated proteoglycan. Molecules of this nature have been detected on the surface of a variety of infectious organisms, and are known to interact with C nonimmunologically. Their location at host-parasite interfaces may have strategic significance in evasion of immune rejection and this possibility is discussed in relation to current understanding of immunity to T. taeniaeformis.
T Cell Tolerance in the Chicken
Authors: Grebenau Mark D. , Thorbecke G. Jeanette
Published: December 2022

T cell-mediated delayed hypersensitivity (DH) to human γ-globulin (HGG) can be induced in chickens by subcutaneous injection of the antigen in complete Freund's adjuvant (CFA). In the present work, it has been demonstrated that specific tolerance of the cells mediating this DH can readily be induced in both normal and bursectomized (BX) FP strain chickens by simple i.v. injection of soluble antigen, regardless of the presence of antibody production to the tolerogen. A significant degree of tolerance at the DH and helper T cell levels could be generated in BX birds by injection of as little as 0.5 mg of HGG; such a dose only induced tolerance in normal birds when it had been previously deaggregated by ultracentrifugation. Regular, nondeaggregated antigen could produce tolerance in normal animals, but only at doses of >5 mg. The tolerizing injection induced a primary antibody response in normal birds in all cases, but a secondary response could not be obtained in animals rendered tolerant at the T cell level.

Establishment of tolerance appeared to be very rapid, and animals remained refractory to induction of DH for at least 3 weeks after the tolerizing injection. The mode in which the antigen was presented to the animals appeared to be crucial in determining whether tolerance or sensitivity would be established.