bluuebunny/crossref_metadata_2025 huggingface dataset:
Evidence for the Presence of Components of the Alternative (Properdin) Pathway of Complement Activation in Respiratory Secretions
Authors: Robertson J. , Caldwell J. R. , Castle J. R. , Waldman R. H.
Published: December 2022

Activation of the alternative complement pathway by respiratory secretory IgA was demonstrated by incubating purified, aggregated preparations of serum and secretory IgA with neat human serum. No depletion of the early components (C1-4) was observed, but 63 and 70% of C3-9, respectively, were consumed. The C3-9-consuming capacity of heat-aggregated nasal secretions from an IgA-deficient volunteer was compared with heat-aggregated nasal secretions from a normal volunteer known to have secretory IgA. The normal nasal secretions consumed C3-9, whereas the IgA deficient secretions did not. Reconstitution of the nasal secretions from the IgA-deficient volunteer with purified secretory IgA produced alternative pathway activation.

Factor B of the alternative complement pathway was found to be present in 16 of 18 bronchoalveolar lavage samples (BALF) from normal volunteers. Simultaneous measurement of lavage and serum albumin and Factor B concentrations rendered it unlikely that Factor B was merely a transudative product from serum in half the samples but rather suggested that it may be a component of lower respiratory tract secretions. The presence of an intact alternative complement pathway in BALF was indicated by showing that cobra venom factor and endotoxin cleaved functionally pure human C3 when mixed with BALF, but had no effect on C3 in the absence of BALF.
An in Vitro Immuno-Enzymatic Assay of Tumor Antigens in the Mouse with β-Galactosidase
Authors: Celada Franco , Giorgio Natali Pier , Radojkovic Jasna
Published: December 2022

A method for detection of the primary binding of soluble tumor-associated antigens by antibodies has been developed by using an enzyme immunoassay (EIA). A heteroantiserum was produced by injecting tumor cells from a chemically induced murine sarcoma into rabbits, and antibodies reacting with most normal tissue components were removed by exhaustive in vivo absorption. A soluble preparation of tumor cells, obtained by 3 M KCl extraction, was conjugated to β-galactosidase from Escherichia coli. The antibody binding was measured by determining the enzyme activity that could be separated by anti-antibody coprecipitation. The reaction follows saturation kinetics, and nonlabeled antigen can be readily quantitated by inhibition. The present method detects determinants common to several MC-induced tumors on the same mouse strain but absent in normal cells and nonrelated tumors in addition to individual tumor-specific transplantation antigens. The sensitivity and simplicity of the new method compare favorably with a binding assay that utilizes radioactive iodine as a label. Thus, EIA becomes a flexible tool for the further characterization and purification of these antigens.
Suppression of Reaginic Antibody Formation
Authors: Lee Weng Y. , Sehon Alec H.
Published: December 2022

Reaginic antibodies to the benzylpenicilloyl determinant (BPO) and ovalbumin (OA) were induced readily in B6D2F1 mice by a single i.p. injection of either 1 or 10 µg of BPO4-OA suspended with 1 mg of Al(OH)3 in 0.5 ml of saline. Administration of conjugates consisting of the hapten coupled to the isologous, nonimmunogenic murine γ-globulins (MγG), i.e., BPO9-MγG, BPO11-MγG, or BPO12-MγG, resulted in complete and specific suppression of the induction of the anti-BPO reaginic antibody response without affecting, however, the level of reaginic antibodies to OA. Further study of the effect of epitope density on the immunologic properties of BPOx-MγG revealed that a) the lightly haptenated conjugates, BPO1-MγG and BPO2.9-MγG, were not immunosuppressive, b) the conjugates, BPO4.3-MγG and BPO19-MγG, were partially tolerogenic, and c) the heavily haptenated conjugate, BPO31-MγG, was nontolerogenic. Moreover, most importantly, the ongoing anti-BPO response in sensitized mice was readily abrogated by either four daily or four weekly injections of BPO9-MγG. The immunosuppressive effect of BPO12-MγG conjugates was dose dependent, complete suppression being achieved with 200 µg of the tolerogen. The unresponsiveness to BPO of spleen cells from immunosuppressed donors was also maintained in adoptive cell transfer experiments in spite of the additional administrationof the immunizing antigen under conditions expected to yield a secondary IgE response. Hence, it is suggested that, with special precautions to prevent unleashing an anaphylactic shock, treatment of penicillin-sensitive individuals with polyvalent conjugates of an appropriate number of BPO groups per human γ-globulin molecule would constitute a rational immunotherapeutic procedure for the abrogation of the allergic response to BPO.
An Appraisal of Fc Receptors on Human Peripheral Blood B and L Lymphocytes
Authors: Lobo Peter I. , Horwitz David A.
Published: December 2022

Human circulating lymphocytes with easily detectable surface immunoglobulin have been divided into two populations, B cells and L cells. This second population lacks membrane-incorporated Ig, but has a receptor for membrane-labile cytophilic IgG. In this study purified B and L lymphocytes were examined for Fc receptors that bind aggregated IgG and IgG complexed to erythrocytes. Purified lymphocyte populations were prepared by nylon columns and by negative selection with rosetting techniques. L lymphocytes bound aggregated guinea pig and human IgG, and formed rosettes with human erythrocytes sensitized with Ripley IgG (EA). Treatment of L lymphocytes with trypsin had no effect on the receptors for IgG. B lymphocytes did not bind EA and attachment of aggregated IgG was variable; up to one-third of these cells fixed aggregated human IgG to the cell membrane. Trypsin treatment abolished binding of Agg-IgG to B cells in sharp contrast to its effect on L cells. Furthermore, double-label immunofluorescence studies showed that cells with both membrane-incorporated Ig and receptors for aggregated guinea pig IgG were rare. These studies indicate that human peripheral blood B lymphocytes lack a high affinity, trypsin-resistant Fc receptor that is present on L lymphocytes.
Replication of Dengue-2 Virus in Cultured Human Lymphoblastoid Cells and Subpopulations of Human Peripheral Leukocytes
Authors: Theofilopoulos Argyrios N. , Brandt Walter E. , Russell Philip K. , Dixon Frank T.
Published: December 2022

We studied the susceptibility of four human lymphoblastoid cell lines (HCL) and of subpopulations of circulating peripheral human leukocytes to dengue-2 virus infection. HCL with B cell characteristics (Raji, Wil 2WT, 8866), B-type peripheral lymphocytes, and macrophages were productively infected by dengue-2 virus. In contrast, an HCL with T cell characteristics (MOLT-4), T type peripheral lymphocytes, and polymorphonuclear (PMN) cells did not become infected and replicate dengue-2 virus. PMN cells did not adsorb dengue-2 virus, suggesting lack of viral receptors. However, T-type cultured lymphoblasts and T-type peripheral lymphocytes adsorbed dengue-2 virus, suggesting that the block in viral replication involves some stage of infection occurring after adsorption. Permissiveness of B-type HCL to dengue-2 virus infection was dependent on the virus seed used but the virus titers obtained among the susceptible HCL varied. HCL infected persistently with dengue-2 virus have been established. Human peripheral lymphocytes inoculated after cultivation for 3 days in complete medium alone or complete medium supplemented with mitogens replicated dengue-2 virus. In contrast, unstimulated peripheral lymphocytes inoculated immediately after isolation adsorbed dengue-2 but did not support its replication. Mitogen-treated and untreated macrophages replicated dengue-2 virus equally well. The efficiency of dengue-2 virus replication by macrophages was higher than that of peripheral lymphocytes but lower than that of HCL.
In Vitro Evaluation of Immunity to Coxiella Burnetii
Authors: Hinrichs David J. , Jerrells Thomas R.
Published: December 2022

The rickettsial organism, Coxiella burnetii, is readily phagocytized by macrophages maintained in vitro. This phagocytosis does not lead to destruction of the organism but rather to intracellular multiplication within the macrophage. Specific antiserum added to the macrophage culture before or after infection, or reacted directly with C. burnetii organisms, fails to control subsequent intracellular replication. Macrophage cultures can be treated with lymphocyte products so that intracellular multiplication by C. burnetii is resisted. These lymphocyte products can be obtained from the culture supernatants of sensitized lymphocytes and antigens or from culture supernatants of Concanavalin A and normal lymphocytes. The activation activity of the lymphocyte supernatants paralleled their content of MIF.
Genetic Control of the Immune Response in Rats to the Known Sequential Polypeptide (Tyr-Glu-Ala-Gly)n
Authors: Luderer Albert A. , Maurer Paul H. , Woodland Robert T.
Published: December 2022

In inbred rats, the antibody response to the known sequential polypeptide (Tyr-Glu-Ala-Gly)n (T-G-A-Gly)n is under the control of two independently assorting loci; (co) dominant, Ag-B-linked Ir-(T-G-A-Gly) I, controlling qualitative responsiveness, and a non-Ag-B-linked modifier locus termed Ir-(T-G-A-Gly) II, controlling the level of antibody produced. The antibody response to (T-G-A-Gly)n was solely IgG and the level of antibody produced was dependent upon Ir-(T-G-A-Gly) II for phenotypic response type.
Depression of Macrophage Function by a Factor Produced by Neoplasms: A Mechanism for Abrogation of Immune Surveillance
Authors: Pike Marilyn C. , Snyderman Ralph
Published: December 2022

The possibility that macrophages mediate surveillance against the development of neoplasms has been receiving increasing support. The acquisition, by neoplastic cells, of the capacity to subvert macrophage function may be an important mechanism by which they escape destruction by the host and become established tumors. Indeed, animals implanted with syngeneic neoplasms developed depressed macrophage migratory ability in vivo and chemotactic responsiveness in vitro. It therefore seemed plausible that neoplasms might be capable of producing inhibitors of macrophage function. The present report describes the identification of such a low molecular weight (6,000 to 10,000), heat-stable inhibitor of murine macrophage accumulation in vivo and chemotaxis in vitro. The inhibitor of macrophages was present in four different murine neoplasms but not present in normal liver, spleen, or inflammatory exudate cells and did not affect PMN chemotaxis in vitro. When given with low numbers of neoplastic cells, the inhibitor increased both the frequency of tumor development and rate of tumor growth. By producing inhibitors of macrophage function, neoplasms may escape initial host surveillance mechanisms.
Independent Effects of IgG and Complement upon Human Polymorphonuclear Leukocyte Function
Authors: Goldstein Ira M. , Kaplan Howard B. , Radin Allen , Frosch Matthew
Published: December 2022

Particle ingestion by polymorphonuclear leukocytes (PMN) is promoted by cell surface recognition and binding of fragments of the third component of complement (C3) and Fc regions of certain immunoglobulin (IgG) molecules. In order to determine the influence of these specific ligand-surface membrane interactions upon other PMN functions, we have employed nonphagocytosable particles (serum-treated Sepharose beads) coated with fragments of C3 and/or IgG, and have investigated whether these provide a sufficient stimulus for the metabolic changes and degranulation that ordinarily accompany phagocytosis by PMN. Sepharose 4B activates complement in fresh normal serum and consequently is coated with fragments of C3 (confirmed by immunoelectrophoretic evidence of factor B and C3 conversion and by immunofluorescence). Adsorbed IgG could be removed from serum-treated Sepharose by boiling in 2 M NaCl without significantly influencing bound complement. We have found that normal human PMN recognize and adhere to Sepharose beads coated with fragments of C3 and consequently are stimulated to increase their oxidative metabolism (measured as superoxide anion generation). This PMN response occurred in the absence of IgG but could be amplified if this immunoglobulin was also present on the bead surfaces. Both adherence and metabolic stimulation could be blocked by treatment of the beads with F(ab′)2 anti-C3. In contrast to metabolic stimulation, degranulation (selective extracellular release of lysosomal constituents) was observed only when PMN encountered both C3 fragments and IgG on the beads. This response could be blocked by treating beads with either F(ab′)2 anti-C3 or F(ab′)2 anti-IgG. These results indicate that cell surface stimulation of PMN is not an “all or none” phenomenon and that certain vital functions of these cells may be mediated or modulated independently by immunoglobulins and complement.
Chemotactic and Bactericidal Capacities of Human Newborn Monocytes
Authors: Kretschmer Roberto R. , Stewardson Prudence B. , Papierniak Cynthia K. , Gotoff Samuel P.
Published: December 2022

We evaluated the chemotactic and bactericidal capacities of human cord blood monocytes, and the ability of cord lymphocytes and sera to generate chemotactic (LDCF) and opsonic factors for monocytes. Our results suggest that the intrinsic locomotive capacity, and the receptor system for LDCF, are adequately developed in the newborn monocyte. Moreover, newborn lymphocytes appear to produce adequate amounts of LDCF, capable of attracting adult monocytes. Poor chemotaxis was observed only when cord monocytes were exposed to supernatants of cord lymphocytes, which suggests that both contribute to this abnormal response. An inhibitory factor for which only cord monocytes have appropriate receptors, or a subtle difference in sensitivity of monocytes and strength of chemotactic factor between adults and newborns would explain these results. Bactericidal capacity of cord monocytes against Escherichia coli K-12 opsonized by either cord or adult serum was comparable to that of adult monocytes.
Mitogen-Stimulated Glutaraldehyde-Fixed Spleen Cells: Ability to Stimulate in the Mixed Lymphocyte Reaction and Generate Effector Cells in Cell-Mediated Lympholysis
Authors: Lightbody James J. , Kong Yi-Chi M.
Published: December 2022

Mouse spleen cells treated with glutaraldehyde lose their stimulating ability in the MLR. If the spleen cells are first converted to a blastogenic state by lipopolysaccharide and subsequently fixed with glutaraldehyde, their stimulating capacity is maintained.
Partial Separation of Human Blood Leukocytes by Density Gradient Electrophoresis: Different Mobilities of Lymphocytes with IgG, Those with IgM and IgD, T Lymphocytes, and Monocytes
Authors: Ault Kenneth A. , Griffith Ann L. , Platsoucas Christos D. , Catsimpoolas Nicholas
Published: December 2022

Previous studies on the electrophoretic separability of human T and B peripheral lymphocytes have produced conflicting results. Vassar et al. (1) using the microscope chamber technique were unable to show any correlation between electrophoretic mobility and degree of enrichment in either T or B lymphocytes. In contrast to the above results, other laboratories (2–6) have shown charge differences between human T and B cells of such magnitude that suggest the possibility of complete separability of these two populations. However, the microscope chamber method does not provide actual physical separation of the electrophoresed cells. Recent attempts by Stein (7) and Häyry et al. (8) to separate physically T and B human lymphocytes by free flow electrophoresis resulted only in partial enrichment of these cells in certain fractions. In consideration of the above findings and in view of the importance of separating human lymphocyte subpopulations by physical methods, we have applied the recently developed technique of density gradient electrophoresis (9, 10), to examine the separability of these cells.
Presidential Address to the American Association of Immunologists, Delivered in Anaheim, California, April 13, 1976
Authors: Good Robert A.
Published: December 2022

I faced this Presidential Address to the American Association of Immunologists with both humility and enthusiasm. Humility toward the responsibility, and enthusiasm for being able to speak without immediate rebuttal. I realized I would speak in the Bicentennial Year … the Bicentennial of our country, which can be viewed also as a Bicentennial of Immunology. It seemed appropriate to present an overview of our surging discipline and to analyze its origin and future. I considered and discarded several titles. “Milestones of Progress in Immunology”, for example, seemed both trite (because this year everyone is speaking of milestones), and also overwhelming. We have come too far too fast to permit comprehensive coverage. I decided to try to present an overview from my personal perspective and to inscribe a series of “runestones” to mark the progress of the exciting travels I see as being particularly important in the development of our discourse. Figure 2A and B are photographs of Runestones.
Interaction of K Lymphocytes with Myeloma Proteins of Different IgG Subclasses
Authors: Spiegelberg Hans L. , Perlmann Hedvig , Perlmann Peter
Published: December 2022

Human myeloma proteins of the four IgG subclasses and their Fc, F(ab)2, and Fab fragments were tested for their ability to inhibit antibody-dependent human K lymphocytemediated cytotoxicity to chicken erythrocytes (CRBC) sensitized with specific rabbit antibodies. In addition, the adsorption of K cells onto glass bead columns coated with myeloma proteins was investigated. Myeloma proteins and their Fc fragments of all four subclasses inhibited K cell activity. However, there were wide variations within a given subclass and IgG2 and IgG4 proteins usually inhibited less than IgG1 and IgG3 proteins. Aggregation of the weakly inhibitory proteins with bis-diazotized benzidine increased their inhibitory effect. An IgG1 half-molecule with a deletion in the Cγ3 domain was weakly inhibitory. Passage of lymphocytes through glass bead columns coated with IgG1 and IgG3 proteins removed K cell activity. In contrast, columns coated with IgG2 and IgG4 proteins, even when aggregated with BDB, failed to absorb K cells but removed significant numbers of SIg positive B lymphocytes. An enhancement of the antibody-dependent cytotoxicity was observed in 34% of the inhibition experiments in the presence of low concentrations of the weakly inhibitory proteins, usually IgG2 and IgG4. This enhancement occurred more frequently (53% of the experiments) with Fc fragments independent of the subclass. Moreover, addition of IgG2 and IgG4 but not IgG1 and IgG3 fragments induced a dose-dependent cytotoxicity to CRBC in the absence of anti-CRBC antibodies. These data indicate that IgG2 and IgG4 proteins have a lower affinity to K cells than IgG1 and IgG3 proteins and are compatible with an earlier hypothesis that proposes that more than one site on the Fc fragment can react with Fc receptors. The present results suggest in addition that there may be functionally different sites, one having a triggering function in K lymphocyte lysis that may be localized on the second constant domain and one being responsible for high affinity binding of IgG to cell receptors that is probably localized on the third constant domain.
Identification of Multiple Cytolytic Components Associated with the β-LT Class of Lymphotoxins Released by Mitogen-Activated Human Lymphocytes in Vitro
Authors: Hiserodt John C. , Fair Daryl S. , Granger G. A.
Published: December 2022

Cytotoxic activity(lymphotoxin (LT) associated with the 50,000 dalton β-LT class of human lymphotoxins has been resolved into multiple components by chromatography on DEAE-cellulose or electrophoresis on polyacrylamide gels. The two components, termed β-LT1 and β-LT2, are separable from the 90,000 m.w. α-LT, since neither component is neutralized by incubation with heterologous rabbit anti-α-LT antisera. However, the two components differ in their overall charge and relative heat stability with the βLT1 component being more positively charged than the β-LT2 molecule and extremely heat labile.
Anti-Immunoglobulin Stimulation of Murine Lymphocytes
Authors: Weiner Howard L. , Moorhead John W. , Yamaga Karen , Kubo Ralph T.
Published: December 2022

Splenic B cells from normal mice, 7 months of age or older, can be stimulated to proliferate in vitro with soluble anti-immunoglobulin (Ig) reagents. To investigate whether the integrity of the anti-Ig molecule is necessary for stimulation and to determine whether cross-linkage of cell surface Ig is required, experiments were done by using F (ab′)2 fragments and Fab monomers prepared from anti-MIgM serum. To determine whether antibodies directed against heavy chains would induce cell proliferation, class-specific antisera were prepared and tested. The results showed that cell proliferation was induced by F (ab′)2 fragments but not by Fab monomers. In addition, cell proliferation was obtained with monospecific antiserum directed against µ heavy chains but not with antisera directed against α- or γ- chains. Thus cross-linkage of µ heavy chains on the B cell surface is required for soluble anti-Ig-induced proliferation. Further experiments were done to investigate the nature of the age-associated response by comparing membrane immunoglobulin density and class on spleen cells from old and young (2 to 3 months) mice; no differences in surface immunoglobulins were found which would explain the age-associated response to anti-Ig reagents.
The T Cell Dependence of B Cell Differentiation Induced by Pokeweed Mitogen
Authors: Keightley Richard G. , Cooper Max D. , Lawton Alexander R.
Published: December 2022

The in vitro plasma cell response of human peripheral blood B lymphocytes to PWM was found to be dependent on the presence of T lymphocytes. B lymphocytes cultured alone with PWM did not respond with proliferation or differentiation into plasma cells. When T lymphocytes were added back to B lymphocytes before culture, the number of plasma cells of the IgM, IgG, and IgA classes formed increased in proportion to the number of T lymphocytes returned. Allogeneic T lymphocytes from either peripheral blood or adult thymus could effectively substitute for autologous T lymphocytes. By comparing responses of B lymphocytes cultured with equal numbers of autologous or allogeneic T cells, it was found that the considerable variation in the magnitude of the plasma cell response from different normal donors was related to a quality of their T cells; T cells from high responders substantially enhanced plasma cell differentiation of B cells from low responders and vice versa. This T cell “helper function” was irradiation-resistant and was a property of thymocytes as well as peripheral T cells.
B Cell Differentiation Induced by Lipopolysaccharide
Authors: Kearney John F. , Cooper Max D. , Lawton Alexander R.
Published: December 2022

Direct immunofluorescence was used to determine the classes of immunoglobulins expressed on the surface membrane and in the cytoplasm of newborn and adult B lymphocytes differentiating in response to LPS in vitro.

In both newborn and adult spleen, a small proportion of IgM-bearing B lymphocytes also stained for IgG2; adult spleen contained an additional population of lymphocytes bearing IgG2 alone. Combined surface and cytoplasmic staining at intervals after culture initiation demonstrated both IgM and IgG2 on the membranes of the earliest cells synthesizing cytoplasmic IgG2. At later stages the proportion of IgM-IgG2 surface doubles and of cells synthesizing cytoplasmic IgG2 which had surface IgM fell significantly. Detection of surface IgM on IgG2 precursors correlated with susceptibility of the IgG2 precursors to anti-µ suppression over the first 3 days in cultures of newborn spleen cells. After 3 days when these cells no longer expressed surface IgM, IgG2 responses were not suppressed although the IgM response was still inhibited. These results suggest that virgin IgG2 precursors may be B lymphocytes expressing both IgM and IgG2, and that “switching” involves the loss of IgM receptors as these cells proliferate and mature.
Relationship of Functional Levels of Early Components of Complement to the H-2 Complex of Mice
Authors: Goldman Margaret B. , Goldman John N.
Published: December 2022

The relationship between the H-2 complex and the complement system was studied by performing functional assays for whole complement and individual complement components on sera from congenic mice of B10 and C3H backgrounds. Levels of whole complement activity were seen to be influenced by S region genes, confirming the work of others. Levels of C3 through C9, however, were independent of the H-2 genotypes suggesting that the differences in whole complement activities were due to variation in levels of C1, C4 or C2. Titrations of these three earliest acting classical complement components were performed on sera from mice of nine strains of the B10 background and four strains of the C3H background. It was shown that functional levels of C1, C4, and C2 were all under control of genes within or very closely linked to the S region.
Relationship between the C5 Peptides Chemotactic for Leukocytes and Tumor Cells
Authors: Romualdez Alberto G. , Ward Peter A. , Torikata Takaatsu
Published: December 2022

A reciprocal relationship has been demonstrated between the generation of the leukotactic and the tumor cell chemotactic peptides from C5. Trypsin-produced C5 leukotactic peptides can be converted into tumor cell chemotactic factors by treatment with tumor cell extracts or by incubation with the α-globulin chemotactic factor inactivator (CFI) isolated from human serum. A similar CFI isolated from rat serum can, like the α-globulin CFI from human serum, generate the tumor cell chemotactic factor from the whole human serum. These results suggest that the tumor cell chemotactic factor derives from the same portion of the C5 molecule as the leukotactic peptide. Furthermore, it appears that the critical change may be cleavage of the amino-terminal portion of the leukotactic peptide. In addition, it has been shown that most normal tissues contain an enzyme-like material that is able to generate in normal human serum a factor chemotactic for tumor cells. The implications of these findings are discussed.
Stimulation of Immune Mechanisms against Mammary Tumors by Incomplete T Cell Depletion
Authors: Roubinian Jirayr R. , Lane Mary-Ann , Slomich Maxell , Blair Phyllis B.
Published: December 2022

When BALB/cfC3H females are subjected to continuous suction thymectomy, a procedure that results in retention of thymic remnants, the latent period before tumor development is significantly prolonged. However, when BALB/cfC3H females are thymectomized by controlled suction, a procedure which removes thymic lobes completely, there is no effect on mammary tumorigenesis. Our results show that incomplete T cell depletion causes premature onset of non-T cell cytotoxicity, an augmentation of T cell cytotoxicity, and an alteration in a serum-blocking activity to mammary tumor target cells as tested in microcytotoxicity assay. On the other hand, complete neonatal thymectomy effectively and completely abrogates immune response to mammary tumor target cells. The inability of completely thymectomized mice to generate an immune response to MTV suggests (but does not prove) that MTV-induced mammary tumor target cell surface antigens are thymus-dependent.
Antibodies to Guinea Pig Lymphokines
Authors: Geczy Carolyn L. , Geczy Andrew F. , de Weck Alain L.
Published: December 2022

An antiserum, (anti-lymphokine globulin, ALyG) directed against highly purified products of activated lymphocytes, inhibits the proliferation of responding cells in guinea pig mixed leukocyte cultures (MLC). This serum recognizes three newly synthesized lymphocyte products (one of them being migration inhibition factor (MIF) which are involved in the mediation of delayed hypersensitivity reactions in vivo. Since ALyG does not appear to contain cytotoxic antibodies against guinea pig lymphocyte antigens and its inhibitory activity cannot be removed by absorption with lymphoid cells it seems likely that the inhibition of MLC reactivity is not mediated by the lysis of stimulator or responder cells and its target is not the lymphocyte per se but possibly some factor elaborated during MLC response. By contrast, antisera with specificity for histocompatibility (H) antigens can inhibit the MLC when the appropriate H antigens are present on the responding and/or stimulating cell population. However, this inhibitory activity of the antisera can be effectively absorbed with lymphoid cells bearing the appropriate H antigens.

The addition of ALyG even 48 hr after the initiation of culture results in a marked inhibition of MLC reactivity. This finding is consistent with the elaboration of a mitogenic factor or signal during the first 48 hr of culture and the delivery of this signal to the responding cell population. Thus, ALyG does not appear to interfere with the synthesis of this factor but must be present after its release in order to block responder cell proliferation. Furthermore, the stimulatory effect of this MLC mitogenic factor, but not of PPD-induced mitogenic factor on “third party” cells can be completely inhibited when the cells are cultured in the presence of ALyG.

These findings suggest that the MLC-mitogenic factor in this system is identical to or cross-reactive with one or both of the newly synthesized molecules recognized by the ALyG.
Analysis of Murine Oncofetal Antigens as Tumor-Associated Transplantation Antigens
Authors: Chism Stanley E. , Wallis Sally , Burton Robert C. , Warner Noel L.
Published: December 2022

In previous studies with in vitro activated cytotoxic T lymphocytes, we have demonstrated the presence of oncofetal antigens (OFA) on a range of murine tumor cells. The present studies with the same tumor lines attempt to determine whether these antigens are also capable of activating lymphocyte responses in vivo. Several experimental designs were followed, each being performed many times (1). Preimmunization of mice with irradiated fetal liver cells and followed by challenge with viable tumor cells did not consistently produce a state of tumor resistance. However, injection of live fetal cells frequently led to enhanced tumor growth (2). The growth of tumors in multiparous mice can be inhibited, in contrast to controls, but this effect was relatively short lived after parturition (3). Preimmunization with fetal cells in vivo did not result in augmented secondary cytotoxic T cell responses in vitro (4). Immunization of mice with irradiated tumor cells frequently led to a state of resistance to the clonal growth of hemopoietic fetal cells, although again the level of resistance was usually relatively weak. From the overall results, we conclude that OFA are relatively poor immunogens on tumor cells or fetal cells in vivo, and in contrast to in vitro responses, do not act as potent tumor transplantation antigens.
Association of Histocompatibility Haplotype HLA-A3-B7 with Multiple Sclerosis
Authors: Bertrams Hans J. , Kuwert Ernst K.
Published: December 2022

Tissue typing was performed on 1000 unrelated multiple sclerosis (MS) patients and 1000 unrelated healthy controls. In addition, segregation analysis of HLA antigens and haplotypes in 36 MS families was done. The following results were observed: HLA-A3 and HLA-B7 were found significantly more often in MS individuals; haplotype HLA-A3-B7 was observed significantly more often in MS patients than in healthy individuals of the same population; the association between HLA-A3 and HLA-B7 was significantly stronger in MS patients than in controls; the haplotype HLA-A3-B7 shows a common segregation with the disease multiple sclerosis (p = 0.02); and the hypothetical “MS-disease-susceptibility-gene” may be located between the second locus of the HLA region and the MLC (mixed lymphocyte culture)-histocompatibility-determinant-region on chromosome No. 6.
Anti-Lymphokine Antibody
Authors: Kuratsuji Tadatoshi , Yoshida Takeshi , Cohen Stanley
Published: December 2022

The present study was designed to explore the range of biologic activity of an anti-lymphokine antiserum (ALKS). This preparation, which was produced by a two-stage immunization procedure, was used to prepare Sepharose bead immunoadsorbent columns. Columns made with ALKS, but not with anti-control antiserum, could specifically remove MIF activity, but not LT, MF, or NCF activity from activated lymphocyte culture supernatants. MIF, but not LT, MF, or NCF could be recovered from the beads by acid extraction. These findings, taken in conjunction with the previously described capacity of ALKS to remove MCF and SRF activity from supernatants, demonstrate that ALKS, although prepared against a lymphokine preparation with multiple activities, has restricted specificity. Thus far, its targets appear to be those mediators that affect macrophages either in vitro or in vivo. The capacity of ALKS to suppress delayed hypersensitivity reactions in the guinea pig is consistent with these observations, since the macrophage is the predominant infiltrating cell in those reactions.
Possible Evidence for Antibody-Dependent Macrophage-Mediated Cytotoxicity Directed against Murine Adenocarcinoma Cells in Vivo
Authors: Haskill J. Stephen , Fett James W.
Published: December 2022

A macrophage-mediated antibody-dependent cellular cytotoxic reaction has been described in a syngeneic murine mammary adenocarcinoma model. The antibody appears to be present in two forms, one apparently complexed with antigen and the other identified as 7S immunoglobulin on Sephadex G-200 and as the IgG2a subclass by blocking of Fc receptors on the macrophages. This antibody is first detected in mice with regressing tumors. Tumor cells from immune mice appear to be coated with this antibody, as they are sensitive to the effects of normal macrophages in vitro, whereas tumor cells from immunologically compromised mice are not. Thus, a macrophage-dependent reaction probably is relevant in the in situ immune defenses against this tumor.
T and B Lymphocytes in the Regulation of Delayed Hypersensitivity
Authors: Neta Ruth , Salvin S. B.
Published: December 2022

A correlation was demonstrated between the transient nature of a) delayed intradermal responses of guinea pigs sensitized to hen egg albumin in incomplete Freund's adjuvant, and b) the proliferative response of sensitized lymphocytes to the specific antigen. Spleen cells from sensitized animals suppressed the proliferative response of lymph node cells to specific antigen. This suppression was dependent on the dose of spleen cells and the time of their removal after sensitization. Thymus cells were suppressive to a lesser extent, and their activity was not correlated with the time of removal after sensitization. Separation of spleen and thymus cells into T and B populations indicated that the B cell was the major suppressor cell in the spleen, while the T cell in the thymus had a similar but less pronounced action.
Relationship Between Homogeneous IgG Fractions and Measles Virus Antibody Activities in Subacute Sclerosing Panencephalitis Brain
Authors: Mehta P. D. , Kane A. , Thormar H.
Published: December 2022

Cellulose acetate and agarose gel electrophoresis of pH 7.4 brain extracts from three patients with subacute sclerosing panencephalitis (SSPE) revealed two to three homogeneous bands in the γ-globulin region. The IgG concentration in the brain extracts ranged from 6 to 8.5% of the total protein. The IgG was isolated by passing each brain extract twice through gel filtration on Sephadex G-200 column. The Sephadex-isolated IgG preparations, at a concentration of 1 mg/ml, had hemagglutination inhibition (HI) titers ranging from 512 to 800, complement fixation (CF) titers of 64 to 100, and neutralizing titers of 400 to 1024 when tested against envelope, nucleocapsids, and whole measles virus, respectively. These IgG preparations were further separated into several fractions by isoelectric focusing (IEF) technique and the reactivities of these eluted fractions were examined against different specific measles virus components and against whole virus. The results showed that both slow and fast moving homogeneous IgG had high HI and neutralizing titers. However, CF antibodies were present only in specific IEF-eluted fractions and their profiles differed somewhat from one brain to another. The IgG fractions reacted with both κ-and λ-chain antisera, but the reactivity varied from one brain to another. When the polyacrylamide gel electrophoretic (PAGE) patterns of heavy (H) and light (L) chains of SSPE IgG were compared to those of IgG from controls, the H chain patterns of both groups were identical whereas the SSPE brain IgG demonstrated two distinct bands in L chain as compared to a single diffuse band observed in controls. These latter results, in addition to homogeneous bands and selective CF antibody activity in IEF eluted IgG fractions from SSPE brain, fulfill some of the criteria of monoclonality.
Activation of Human B Lymphocytes
Authors: Fauci Anthony S. , Pratt Karen R. K. , Whalen Gail
Published: December 2022

A recently described system for the induction and assay of plaque-forming cells (PFC) following polyclonal stimulation of human B lymphocytes was employed to delineate the cellular requirements for pokeweed mitogen (PWM)-induced activation of human tonsillar and peripheral blood B lymphocytes. It was found that the process was not dependent on the presence of monocytes, but was dependent on the presence of T cells. T cell depleted suspensions of tonsillar as well as peripheral blood lymphocytes had markedly diminished if not absent PFC responses following PWM stimulation. The PFC responses of T cell depleted cultures of tonsil and blood could be reconstituted by adding back T cells. T cell supernatants could also reconstitute the PFC response of T cell depleted tonsillar lymphocytes but not of T cell depleted blood lymphocytes, thus suggesting a more stringent T cell requirement of the PWM-induced PFC response of blood lymphocytes. Enrichment for T cells on the other hand enhanced PFC responses in both organs, particularly in those cultures which had low responses in unfractionated suspensions demonstrating the requirement of optimal T cell help for maximal PFC responses. This study demonstrates the feasibility and effectiveness of this PFC system in elucidating some of the complex regulatory processes associated with the activation of human B lymphocytes.
Idiotypes of Inulin-Binding Antibodies and Myeloma Proteins Controlled by Genes Linked to the Allotype Locus of the Mouse
Authors: Lieberman R. , Potter M. , Humphrey W. , Chien C. C.
Published: December 2022

The serum from non-immunized mice of strains BALB/c, C58, A/He, and RIII contained hemagglutinins for stearoyl inulin-coated SRBC. Immunization with bacterial levan slightly elevated these titers. These same sera also carried cross-specific idiotypic determinants (IdX) that are associated only with inulin-binding myeloma proteins (INBMP) of BALB/c mice. Three InuIdX specificities, A, B, and G, were identified. The InuIdX phenotypes of strains BALB/c, C58, and A/He were InuIdXA+B+G+; strain RIII was InuIdX A+B-G+; strain C57BL/6, C57BL/10, DBA/2, AKR and NH were A-B-G-. Strains CBA, C3H, PL, and C57L could not be typed because of low and inconsistent levels of InuIdX and anti-inulin hemagglutinins.

The InuIdXA+B+G+ phenotype was used as a genetic marker in immunoglobulin congenic strains CB-20, BAB-14, and BC-8 and in Bailey RI strains which are derived from crosses of BALB/c (InuIdXA+B+G+) and C57BL/ka or C57BL/6, respectively (InuldXA-B-G-). Linkage of the IdXA+B+G+ to the BALB/c a1 allotype locus was demonstrated. In addition, the InuldXA+B+G+ marker was used as a phenotype in an analysis of 168 first generation backcross progeny (C57BL × (C57BL × BALB/c) F1). Linkage of the marker to the BALB/c allotype was found again. Two proven recombinant mice having the C57BL a2 allotype and the InuIdxA+B+G+ markers were identified and progeny tested. Four other potential crossover types are still being progeny-tested.
Modification of H-2 Antigenic Sites by Enzymatic Treatment Influences Virus-Specific Target Cell Lysis
Authors: Ertl Hildegund C. J. , Koszinowski Ulrich H.
Published: December 2022

Vaccinia virus-infected cells were treated enzymatically to remove H-2 antigenic sites. The effect of this procedure on virus-specific cell-mediated cytolysis (CMC) and virus-specific antibody-mediated cytolysis (AMC) was tested. Due to the inhibition of cellular proteinsynthesis by the vaccinia virus infection, H-2 antigenic sites were not resynthesized while there was a continuous production of viral surface antigens.

These cells with a high concentration of viral surface antigens and decreased H-2 determinants could be used as targets in the virus specific AMC. But they were not lysed in the virus specific CMC which emphasizes the significance of H-2 antigens during recognition of virus-specific determinants by T cells.
Mouse B and T Lymphocyte Responses to Purified Timothy Pollen Antigens in Vitro
Authors: Fairchild Sally S. , Malley Arthur
Published: December 2022

Purified populations of splenic B and T lymphocytes from LAF1 mice immunized with a crude extract of timothy pollen (WST) responded specifically to pollen antigens in an in vitro lymphocyte transformation system. The peak lymphocyte transformation response occurred 5 days after a secondary immunization and was the result of T-B cell cooperation in vitro. With two purified pollen antigens as in vitro stimulants we were able to define at least two antigen-specific populations of B cells and one population of T cells. These results were confirmed by inhibition studies with a monovalent hapten from WST, Antigen D.
Experimental Autoimmune Thyroiditis in the Guinea Pig: Characterization of Infiltrating Lymphocyte Populations
Authors: Paget Stephen A. , McMaster Philip R. B. , van Boxel John A.
Published: December 2022

Experimental autoimmune thyroiditis was induced in outbred guinea pigs by isoimmunization with thyroid extract in complete Freund's adjuvant. A digestion procedure using collagenase and deoxyribonuclease was used to make viable single-cell suspensions of pooled thyroid glands from groups of animals with advanced degrees of thyroiditis. Thymus-derived or T lymphocytes, identified by their capacity to form E rosettes with rabbit erythrocytes, were found to be the predominant (75%) infiltrating lymphocyte; bone marrow-derived or B cells consitituted most of the remainder. The infiltrates of inbred animals with thyroiditis were demonstrated to contain cells capable of mediating antibody-dependent lymphoid cellmediated cytotoxicity.
Isolation and Characterization of Guinea Pig Properdin
Authors: Nicholson Anne , Austen K. Frank
Published: December 2022

Guinea pig properdin was purified to homogeneity by employing as an assay during isolation its capacity to augment the hemolytic activity of a heterologous human C3b-dependent C3 convertase, C. The purified protein elicited a monospecific antibody response in a rabbit. The antiserum, by immunodiffusion, gave a reaction of identity between a protein in whole guinea pig serum and the immunogen. A solid phase immunoadsorbent prepared with the antiserum removed properdin function from the purified protein. The purified guinea pig protein exhibited the classical properdin function of reconstituting a human RP for zymosan-induced C3 inactivation. The guinea pig properdin also agglutinated red cell intermediates bearing either guinea pig or human C3b and retarded the decay of homologous C3 convertase, C. These activities are the same as those observed for purified human properdin and validate the amplification function of properdin on terminal component activation in a second species.
Immunologic Reactivity of the Lung
Authors: Hunninghake Gary W. , Fauci Anthony S.
Published: December 2022

The effects of various in vitro and in vivo regimens of corticosteroid administration on guinea pig alveolar macrophages were studied. Corticosteroid-induced immunosuppression was assessed by the effect of drug administration on the total numbers and functional capabilities of alveolar macrophages as measured by the PHA-induced and antibody-dependent cellular cytotoxicity assays against sheep red blood cell targets. In vivo administration of either hydrocortisone sodium succinate (100 mg//kg/one dose) or cortisone acetate (100 mg/subcutaneously for 7 days) caused a marked increase in the numbers of alveolar macrophages recovered from the teased lung cell suspensions at 4 and 24 hr, respectively, after the last injection. Both regimens of corticosteroid administration cause similar levels of peripheral blood lymphocytopenia and monocytopenia, 4 and 24 hr, respectively, after the final injection. Neither in vitro hydrocortisone (0, 1, and 10 µg/ml), nor hydrocortisone (100 mg/kg), in vivo had any effect on either the PHA-induced or antibody-dependent cellular cytotoxicity of alveolar macrophages. In marked contrast, cortisone acetate, depo-preparation which gives sustained elevations of plasma cortisol levels similar to those found for a brief period after i.v. injection of hydrocortisone caused a marked decrease in cytotoxic effector function of alveolar macrophage suspensions. In a separate experiment, the suppressed killer cell function of the alveolar macrophages from steroid-treated animals was found not to be related to an intrinsic defect in killing of bound target cells since the defect in killing could be overcome by increasing the density of antibody and PHA on the target cells.
Interaction of DNP-Antigens with the First Component of Complement (C1̄)
Authors: Loos Michael , König Wolfgang
Published: December 2022

Evidence is presented that DNP-substituted human serum albumin (DNP-HSA) interacts with the first component of complement (C1̄). Fluid phase C1̄ was 60 times more sensitive to DNP-HSA than C1̄ bound to EAC4 (EAC1̄4). The inhibition of C1̄ was strongly dependent on the DNP substitution with optimal activity at approximately 60 DNP groups per HSA molecule. With higher numbers of DNP groups (64 to 68) no further increase of the inhibitory effect on C1̄ was evident. DNP-HSA had no effect on the esterolytic activity of C1̄s the consumption of C4 was not affected by highly substituted DNP-HSA. DNP-HSA, however, was able to precipitate C1q, indicating a direct interaction of DNP-HSA with C1q. The degree of C1q precipitation was strongly dependent on the DNP-HSA concentration. This behavior of DNP-HSA is similar to the interaction of LPS with C1̄ and differs from the C1̄-interaction of polyanions (i.e., dextran sulfate, polyvinyl sulfate, polyanethol sulfonate, etc.) where in addition to binding of C1q, the consumption of C4 by C1̄s is inhibited.
Conformational Integrity of Myoglobin after Immunization with Freund's Adjuvant
Authors: Smith John A. , Hurrell John G. R. , Leach Sydney J.
Published: December 2022

The use of Freund's complete adjuvant for immunizing goats with myoglobin produces mainly antibodies directed against antigenic determinants present in the native protein. Only about 9% of the total antibodies produced are directed toward determinants not expressed in the native molecule. This shows that neither emulsification nor the subsequent in vivo events leading up to the immune response appreciably perturb the conformation of the protein surface.