bluuebunny/crossref_metadata_2025 huggingface dataset:
Human Transfer Factor: Fractionation and Biologic Activity
Authors: Burger Denis R. , Vandenbark Arthur A. , Daves Doyle , Anderson William A. , Mark Vetto R. , Finke Patricia
Published: December 2022

Human transfer factor (TF) was fractionated by exclusion chromatography and the fractions were tested for biologic activity in vivo and in vitro. Specific TF activity in vivo was found to reside in the major UV-absorbing peak (Fraction III). Fraction III eluted at 2.7 × V0 and transferred tuberculin, candida, or KLH-reactivity to previously negative recipients. Fraction III from nonreactive donors was ineffective.

When the fractions were tested in vitro, we found that both the mitogenic activity of whole TF and the suppressive activity to mitogen activation when present in TF was found in Fraction I. Fraction III contained components responsible for augmentation of PHA and PWM responses. In addition, Fraction III contained the component responsible for antigen-dependent augmentation of lymphocyte transformation. Fraction IV was suppressive to antigen-induced lymphocyte transformation. These data suggest that TF preparations contain components which can affect immune reactions in both specific and nonspecific ways.
Tolerance and Contact Sensitivity to DNFB in Mice
Authors: Moorhead John W.
Published: December 2022

Tolerance in contact sensitivity to DNFB can be adoptively transferred to normal mice with lymph node cells from tolerant donors. This tolerance is antigen specific and is mediated by T cells, i.e., “suppressor” T cells. Experiments were carried out to investigate the mechanism(s) by which the suppressor T cells induce tolerance to DNFB contact sensitivity. The suppressor cells were effective only if they were present during the early stages of the afferent limb of sensitization. As measured by DNA synthesis, cell proliferation in the draining lymph nodes of recipients of suppressor cells was found to be significantly less than in control animals indicating that the suppressor cells acted, at least in part, by limiting or inhibiting DNFB-induced cell proliferation. This inhibition was shown to be antigen specific since the DNFB suppressor cells did not inhibit cell proliferation induced by oxazolone, an unrelated contact sensitizer. The ability to DNFB tolerant cells to block afferent sensitization pathways differs from the mechanism of tolerance to picryl chloride, reported by others, where efferent pathways are blocked.
Different Mechanisms for the Modulation of TL Antigens on Murine Lymphoid Cells
Authors: Esmon Naomi L. , Russell Little J.
Published: December 2022

An indirect radioimmunoassay has been used to study the antibody-induced changes in expression of TL antigens on mouse thymus and leukemia cells. The results obtained indicate that the incubation of TL+ cells in relatively high concentrations of anti-TL antisera results in a detectable, but not major, loss in the quantity of antigen detected in the plasma membranes. This decrease is not inhibited by azide. Although the antigen loss seems to coincide with the appearance of patching of membrane TL antigens on leukemia cells observed in indirect immunofluorescence microscopy, this is not the case when thymocytes are used as targets. Fab fragments prepared from anti-TL antibodies can also induce a quantitative decrease in antigen expression. The major mechanism involved in membrane antigen loss seems to be endocytosis, although some shedding may also occur. Resistance to immune cytolysis (antigenic modulation) occurs during incubation in anti-TL antibodies before either endocytosis or antigen redistribution has progressed enough to be directly responsible. Since antigenic modulation does not represent a depletion in TL antigens being expressed on the cell surfaces, another mechanism must account for the failure of guinea pig complement to achieve cell lysis.
Site of Action of a Soluble Immune Response Suppressor (SIRS) Produced by Concanavalin A-Activated Spleen Cells
Authors: Tadakuma Takushi , Pierce Carl W.
Published: December 2022

The cellular site of action of SIRS, a soluble immune response suppressor released by Con A-activated spleen cells which suppresses antibody responses to heterologous erythrocytes by murine spleen cells in vitro, was investigated. Exposure of spleen cells to SIRS for 2 hr at 37°C or 1 hr at 4°C was sufficient to suppress 5-day antibody responses in vitro. Similar exposure of splenic or peritoneal exudate macrophages to SIRS also suppressed antibody responses by untreated splenic lymphoid cells; exposure of splenic lymphoid cells to SIRS was without effect. SIRS did not act via T cells which might have contaminated the macrophage preparations. SIRS-mediated suppression could be partially overcome by an excess of normal peritoneal exudate macrophages, but not by an excess of T or B cells. These data indicate that the target cell of SIRS activity is the macrophage. The results are discussed in the context of macrophage functions that could be affected by SIRS.
Studies of Idiotypic Antibodies: Reactions of Isologous and Autologous Anti-Idiotypic Antibodies with the Same Antibody Preparations
Authors: Scott Rodkey L.
Published: December 2022

Outbred rabbits were immunized with keyhole limpet hemocyanin diazotized to p-aminophenyl-N-trimethylammonium chloride (TMA). Anti-TMA antibodies were specifically purified and the F(ab′)2 fragments were isolated after pepsin digestion. Glutaraldehyde-polymerized anti-TMA F(ab′)2 fragments were reinjected back into the same animal that synthesized the antibodies to elicit an autologous anti-idiotypic (AAI) antiserum and were also injected into an allotypically matched recipient to elicit isologous anti-idiotypic (IAI) antiserum.

AAI and IAI antisera reactive with the same F(ab′)2 preparation were compared by assaying the populations of anti-TMA molecules reactive with each antiserum. The assays showed that the AAI antiserum reacted with smaller populations of molecules than did the IAI antiserum. The population of molecules recognized by the AAI antiserum was contained within the larger population recognized by the IAI antiserum. Hapten inhibition studies showed that the differences in the populations of molecules recognized by each antiserum were not related to the hapten-binding properties of the idiotypic antibodies: in one case both AAI and IAI reactions were equally inhibited by hapten; in another, the AAI reaction was inhibited more than was the IAI; in another, the IAI reaction was inhibited more.
Characterization of T and B Antigen-Binding Cells for β-Galactosidase
Authors: Swain Susan , Coons Albert H.
Published: December 2022

Antigen-binding cells to β-galactosidase were enumerated in thymus and spleen of mice of different ages and found to remain at a constant frequency throughout life. Thymic β-galactosidase-binding cells (ZBCs),6 representing T-binding cells, showed no appreciable fluctuation, whereas splenic ZBCs, a mixture of T- and B-binding cells, were slightly depressed at birth but found at normal frequencies by 1 week of age. In addition, germfree mice, both within the 1st week after birth and after maturity, had nearly as many binding cells as did conventionally reared age-matched mice. These results considered together suggest that the ability of cells of both T and B origin to recognize antigen, as revealed by their ability to bind β-galactosidase, arise independently of specific or nonspecific antigenic stimulation, as part of the normal ontogenic sequence of steps of differentiation. This is consistent with the theory of clonal precommitment of T and B cells.
Metabolic Studies of Guinea Pig Basophilic Leukocytes in Short-Term Tissue Culture
Authors: Galli Stephen J. , Galli Anne S. , Dvorak Ann M. , Dvorak Harold F.
Published: December 2022

Mature circulating guinea pig basophils, purified to comprise 25% or more of leukocytes, have been successfully maintained in short-term tissue culture for up to 72 hr. These cells were found to retain the ability to synthesize histamine, as assayed by a new isotopic-thin layer chromatographic assay which can reliably detect as little as 0.5 pg of 3H-histamine.

Cell-associated, newly synthesized histamine was detectable as early as 1 hr of culture, was substantially increased at 6 hr, and reached maximal levels at 24 hr, when it accounted for approximately 6.5% of total cell histamine. Newly synthesized histamine was still detectable at 48 and 72 hr of culture. Histamine synthesis was decreased by lowering the concentration of histidine in the culture medium, and was markedly reduced by all of the specific histidine decarboxylase (HDC)3 inhibitors tested, but not by α-methyl-DOPA, pyrilamine maleate, or metiamide. Increasing the concentration of pyridoxal phosphate, the HDC coenzyme, above that normally present in culture medium resulted in only an equivocal increase in the amount of newly synthesized histamine, whereas aminoguanidine, an inhibitor of histaminase, had no detectable effect. Uptake of exogenous histamine by cultured basophils was trivial compared to histamine synthesis.

Both newly synthesized and previously manufactured, nonisotopic, histamine seemed to be stored in the same pool, as the same proportion of both was released by concanavalin A (Con A). Cellular histamine was largely conserved, with little or no spontaneous release into the medium of detectable isotopic or nonisotopic histamine.

These techniques provide a model for studying granulocyte metabolic processes in vitro, and should assist in the direct investigation of a variety of their physiologic functions.
Distinct Target Determinants on Two Lymphoblastoid Lines Derived from the Same Individual
Authors: Koide Yukio , Takasugi Mitsuo
Published: December 2022

Specifically cytotoxic cells were generated in unidirectional mixed leukocyte culture against both T and B lymphoblastoid cell lines. The cytotoxicity observed included both specific and less selective components which were differentiated by the interaction analysis based on the two-way analysis of variance. The results indicate that a T cell line and a B cell line derived from the same person possessed distinct target specificities. The antigens detected by direct cytotoxicity were confirmed by inhibition studies with competitor cells.
Functional Heterogeneity Among the T-Derived Lymphocytes of the Mouse
Authors: Araneo Barbara A. , Marrack Philippa C. , Kappler John W.
Published: December 2022

The kinetics of the proliferative response and the appearance of effectors of helper activity after stimulation by antigen were examined in T cell subpopulations. As defined in previous papers of this series, one population, T1, is short-lived after adult thymectomy (ATx), and relatively resistant to elimination by anti-thymocyte serum (ATS). Another population, T2, is long-lived after ATx, but highly sensitive to elimination by small doses of ATS.

From precursors within the T2 population, effectors of specific helper activity, after priming with antigen, appeared within 1 to 2 days and reached a maximum on day 4. The responding cells reached their peak proliferative response within 24 hr after stimulation by antigen. In contrast, helper activity arising from T1 precursors first appeared on day 3 and peaked on day 5. These cells did not reach their maximal proliferative response until 60 hr after priming. These findings indicate additional useful markers for distinguishing the T1 and T2 subpopulations and are consistent with models for T cell development in which T1 cells are virgin cells and T2 cells are memory cells.
Comparison of the Pyrogenicity, Limulus Activity, Mitogenicity and Complement Reactivity of Several Bacterial Endotoxins and Related Compounds
Authors: Elin Ronald J. , Sandberg Ann L. , Rosenstreich David L.
Published: December 2022

The correlation of mitogenicity, complement activation, pyrogenicity and Limulus amebocyte lysate (LAL) gelation induced by 17 bacterial endotoxins (LPS), modified endotoxins, related compounds, and other B cell mitogens was examined. With the use of these compounds, a significant correlation was found between pyrogenicity and mitogenicity (r = 0.85, p < 0.0005), pyrogenicity and LAL reactivity (r = 0.84, p < 0.0005), and mitogenicity and LAL reactivity (r = 0.72, p < 0.0025). In contrast, complement activation by these compounds did not correlate with mitogenicity, pyrogenicity or LAL reactivity (r = 0.09, p τ 0.4; r = -0.17, p τ 0.3; r = 0.17, p τ 0.3, respectively). These results suggest that a specific chemical configuration may induce mitogenicity, pyrogenicity, and LAL reactivity which differs from that responsible for complement activation.
Primary in Vitro Sensitization of Murine Lymphocytes against Isogeneic and Allogeneic Cells Transformed by Simian Virus 40
Authors: Maki Takashi , Howe Michael L.
Published: December 2022

Primary in vitro sensitization of murine lymphocytes to isogeneic and allogeneic cells transformed by simian virus 40 (SV40) is described. The results of specificity studies utilizing cytotoxic effector lymphocytes obtained by in vitro immunization indicate that SV40 transformation results in the expression of tumor-specific antigens which are recognized by cytotoxic effector cells. Moreover, the studies demonstrate that expression of tumor-specific antigens on transformed cells is associated with a reduction in the functional expression of normal histocompatibility antigens.
Genetic Control of Ligand-Induced Events in B Lymphocytes
Authors: Fram Robert J. , Sidman Charles L. , Unanue Emil R.
Published: December 2022

The early cycle of events which follows the binding of anti-Ig ligands to the surface Ig molecules of B lymphocytes was compared in several inbred strains of mice. Kinetic differences were found in the rate of surface Ig capping and absolute differences in extent were found for anti-Ig-induced B cell motility. These strain differences were not restricted to the B cell surface Ig molecules, however, since capping by Con A of its receptor sites on B and T cells and random T cell motility were also affected. A genetic analysis of A and CBA mice revealed that each of the traits studied is probably affected by two gene differences between the two strains analyzed.

The possible roles of microtubules and cyclic nucleotides in these genetic differences were also explored. Colchicine, which depolymerizes microtubules, raised the low capping and motile responses of CBA spleen cells to the high responses characteristic of A strain cells. This implies an excess of microtubular function in CBA cells. Raising the level of cyclic AMP also raised the low CBA capping response to the high A level, but did not differentially effect the motile responses of cells of the two strains. The possible significance and locus of action of these effects is discussed.
Cell Membrane Antigen Isolation with the Staphylococcal Protein A-Antibody Adsorbent
Authors: Kessler Steven W.
Published: December 2022

Procedures are detailed for the rapid isolation of representative cell membrane antigens with protein A-bearing staphylococci as an adsorbent for IgG antibodies complexed with the antigens. Cell surface membrane proteins were radioiodinated and solubilized in nonionic detergent. Specific antisera were subsequently added and the immune complexes precipitated by addition of the staphylococcal adsorbent and low speed centrifugation. The antigens isolated included surface immunoglobulins from mouse and human lymphocytes, human β2-microglobulin and HL-A alloantigens, mouse H-2 alloantigens, and the murine leukemia virus glycoprotein gp 70. Rabbit, sheep, goat, and mouse antisera were all effective for the specific phase of the precipitation reaction. The surface membrane immunoglobulins of mouse splenic lymphocytes and human peripheral blood lymphocytes differed with respect to class composition and protein A reactivity. Mouse lymphocyte surface immunoglobulins were nonreactive with protein A, whereas a high proportion of human lymphocyte surface immunoglobulins of different classes bound directly to the staphylococci. In sequential immunoprecipitation studies the prior isolation of one antigen had no appreciable effect on the subsequent recovery of another antigen. Adsorption of antigen-antibody complexes is quantitative when protein A sites are provided in excess over antiserum IgG sites, and this obviates the need for equivalence point titrations for optimal precipitation necessary with alternative double antibody techniques.
Characterization of a Constitutive Variant of the Murine Serum Protein Allotype, Slp
Authors: Hansen Ted H. , Shreffler Donald C.
Published: December 2022

Properties of the Ss serum protein (putatively the murine homologue of the fourth component of complement) and its allotypic form, Slp, were studied in a wild-derived variant, Ssw7. The Ssw7 allele is unique in that it confers non-sexlimited expression of the Slp allotype. During examination of the H-2 haplotypes of two congenic lines which derived their Ssw7 allele from a common origin, it was found that intra-H-2 recombination had occurred during the establishment of one of these lines. The sex-limited expression of Slp as determined by the Ssd allele was constrasted in several ways to the constitutive expression of Slp as determined by the Ssw7 allele. Slp expression in mice carrying the Ssd allele was found to be subject to control by the testicular feminization mutant, Tfm, while in mice carrying Ssw7 the Slp expression was independent of Tfm control. This implies that Slp is subject to a rather fundamental hormonal control mechanism in mice carrying Ssd and presumably all other Slp-positive Ss alleles, with the exception of Ssw7. It is proposed that Ssw7 may represent an “operator-constitutive” mutation of the Jacob-Monod type.

Including the Ssw7 allele, there are now at least seven distinguishable Ss variants. Each of these Ss alleles is characterized by a unique level of Ss and/or Slp antigenic expression. The sex-limitation difference and quantitative differences in Ss and Slp expression associated with the Ssw7 allele were found to be codominant or intermediate when Ssw7 was in heterozygous combinations with other Ss alleles. This indicates a lack of trans effects and argues against genetic differences in regulatory proteins as the basis for these polymorphisms.
Mapping of the Genetic Control of Murine Response to Low Doses of the Dinitrophenyl Conjugates of Ovomucoid and Bovine γ-Globulin
Authors: Freed John H. , Deak Beverly D. , McDevitt Hugh O.
Published: December 2022

The antibody responses in mice to low doses of the DNP-conjugates of ovomucoid (OM) and bovine γ-globulin (BGG) were measured for a number of inbred strains carrying independent and recombinant H-2 haplotypes. This permitted mapping the gene(s) controlling the response to low doses of OM (Ir-l-OM) within the I-A subregion of the H-2 complex. Similarly, it was possible to map the primary gene(s) controlling the response to low doses of BGG (Ir-l-BGG) within the I-A and/or I-B subregions. Further localization of the Ir-l-BGG gene(s) to the I-A or I-B subregion of the H-2 complex was not possible due to the ambiguous response of the B10.A(4R) strain of mice.
The Induction of Hapten-Specific T Cell Tolerance by Using Hapten-Modified Lymphoid Cells
Authors: Miller Stephen D. , Claman Henry N.
Published: December 2022

BALB/c mice were made tolerant to the T cell-dependent phenomenon of contact sensitivity to DNFB by i.v. injection of syngeneic lymphoid cells which had been previously modified with DNFB in vitro. The highly efficient unresponsiveness, as measured by ear challenge and in vitro antigen-induced cell proliferation, was shown to follow dose-response kinetics both in vivo and in vitro and to be exquisitely specific for the DNP moiety. The kinetics of tolerance induction were shown to be very rapid and had been previously shown to be long lasting. Unresponsiveness was more efficient when the hapten-modified cells were introduced by the i.v. route and tolerance could be increased by repeated injection of tolerogen. The tolerance could be transferred to normal syngeneic recipients by spleen and/or lymph node cells from tolerant donors. A wide variety of hapten-modified lymphoid cells, including mixed cell populations and enriched populations of T cells, B cells, and macrophages, were capable of inducing tolerance. The unresponsiveness was dependent merely on the association of DNP to the lymphoid membrane proteins and not upon the viability of the hapten-modified cells.

These experiments support the hypothesis that in hapten-specific T cell sensitivity, as exemplified by contact sensitivity to DNFB, specific T cell tolerance in actively induced by hapten on self-membrane. In other hapten-specific antibody-forming systems, tolerance appears to be most readily induced by hapten on soluble self protein or on a nonimmunogenic carrier.
The Induction and Regulation of Guinea Pig B-Lymphocyte Proliferation in Vitro
Authors: Lipsky Peter E. , Rosenthal Alan S.
Published: December 2022

Conditions for the in vitro induction of proliferation have been examined in a population of guinea pig lymph node lymphocytes enriched for B lymphocytes. Known B cell mitogens such as bacterial lipopolysaccharide and tuberculin-purified protein derivative induced DNA synthetic responses in this B lymphocyte population as did a number of dinitrophenyl hapten-protein conjugates, all in the apparent absence of T cell participation. B cell proliferation occurred most efficiently in serum-free medium with both heterologous and homologous sera inhibiting maximum responses. In vivo immunization with hapten-protein conjugates failed to enhance subsequent in vitro B cell responsiveness to these materials beyond that owing to recruitment by contaminating antigen-primed T cells. Macrophage-associated antigen, which efficiently triggered T cell proliferation, was much less effective at initiating B cell DNA synthesis than was soluble antigen not bound to macrophages. Furthermore, although accessory cells were required for the development of T cell DNA synthetic responses, macrophages or factors produced by them inhibited spontaneous or induced B cell 3H-thymidine incorporation.
Immunogenicity of Fab Fragment of Protein-315 for BALB/c Mice
Authors: Tungkanak Rachaneepas , Sirisinha Stitaya
Published: December 2022

Immunogenicity of Protein-315 and its Fab fragment (Fab-315) for isologous and heterologous strains of mice was investigated by comparing the characteristics of the anti-idiotypic response produced in BALB/c and A/J mice. The ability of these anti-idiotypic antisera to compete with DNP-lys for the ligand-binding site of Protein-315 were analyzed by comparing their hapten inhibition curves.

The anti-idiotypic responses of BALB/c mice to Protein-315 and to Fab-315 were similar to one another with regard to antibody specificity and sensitivity to inhibition by hapten, suggesting that both forms were equally immunogenic in inbred BALB/c mice. This observation indicates that the Fc portion of Protein-315 is not essential for the induction of anti-idiotypic response. Both BALB/c and A/J mice recognized the same antigenic determinants on Fab-315 since the anti-idiotypic antibodies produced by these animals were indistinguishable with regard to their interaction with Protein-315, Fv-315, and sensitivity to hapten inhibition.

On the other hand, the response of A/J mice to Protein-315 was remarkably different from that of BALB/c mice since it was found that Protein-315 seemed to be more immunogenic in A/J mice than in BALB/c mice. The data also indicated that Protein-315 was more immunogenic than Fab-315 in stimulating the anti-idiotypic response in A/J mice. A/J antiserum to Protein-315 was found to have antibodies specific for the ligand-binding site as well as those specific for other antigenic determinations on the Fc portion. These findings suggest that the Fc portion of Protein-315 may either act as a haptenic determinant to provoke directly the antibody response or as a carrier determinant to enhance the anti-Fab response in A/J mice.
Complement Receptor Lymphocytes in the Rabbit
Authors: Green William R. , Fanger Michael W.
Published: December 2022

Complement receptor lymphocytes (CRL) were detected in various rabbit lymphoid tissues by the ability of these CRL to form rosettes with sheep red blood cells coated sequentially with rabbit antiserum directed against sheep red blood cell stroma and horse serum as a nonhemolytic source of complement (EAC). The rosette assay was shown to be specific for complement receptor (CR) activity and the EAC capable of detecting both C3b and C3d specific receptors. With lymphocyte preparations containing less than 5% phagocytic cells, the average per cent CRL in the various tissues studied was as follows: thymus 1%, popliteal lymph node 18%, spleen 30%, appendix 35%, and peripheral blood 45%, Double assays in which the lymphocytes were prestained with an FITC-labeled Fab′ fragment of a goat anti-rabbit Fab antibody before rosetting indicated that CRL were a subpopulation of surface immunoglobulin (SIg)-bearing lymphocytes in popliteal lymph node, spleen, and peripheral blood. In the appendix, however, in addition to finding SIg+ CR+ and SIg+ CR- populations, an SIg- CR+ population was consistently found. Double assays employing FITC-labeled goat antibodies specific for µ, α, and γ determinants were also performed to determine if there was any relationship between the class of Ig displayed and presence of CR. It appeared that an approximately equivalent percentage of both IgM- and IgG-bearing cells also displayed CR. Experiments in which appendix cells were treated with Pronase to remove SIg and CR and the cells cultured in vitro to allow regeneration of surface markers confirmed the existence of SIg+ CR+, SIg+ CR-, and SIg- CR+ lymphocyte subpopulations. Whether the SIg- CR+, population represents a developing B cell population which will eventually also express SIg or whether it belongs to the T or “null” cell populations is unclear at present.
Role of T Lymphocytes in Adjuvant Arthritis
Authors: Kayashima Kohji , Koga Toshitaka , Onoue Kaoru
Published: December 2022

Rats of Wistar King Aptekman (WKA) were thymectomized at 4 weeks of age and injected with Mycobacterium tuberculosis wax D 6 weeks after the operation to induce adjuvant arthritis. The development of this disease was strikingly enhanced by this treatment. Further experiments showed that a 4- to 6-week interval between thymectomy and wax D injection was necessary to show the enhancing effect. Such enhancement by thymectomy was also shown in rats of another strain, Sprague-Dawley (SD). The enhancing effect of thymectomy was abolished when thymocytes of normal syngeneic rats were transferred to thymectomized rats 7 days before the wax D inoculation. Furthermore, severe arthritis was also produced in WKA rats that were pretreated with low dose (200 R) whole body irradiation, but not in those treated with higher doses (400 to 700 R). These results seem to indicate that the enhancing effect is brought about by selective depletion of a certain population of T lymphocytes. The population depleted may be thymus dependent, short-lived and radiosensitive, the properties of which agree with those known for suppressor T lymphocytes. Thus, it appears that thymus-derived cells could normally exert a regulatory effect on the development of adjuvant arthritis which might render these rat strains relatively less susceptible to this disease.
Prearmed Effector Cells and the Target Cell Specificity of Lectin-Induced Cellular Cytotoxicity
Authors: MacDermott Richard P. , Nash Geoffrey S. , Boldt David
Published: December 2022

We have investigated mechanisms by which plant lectins induce human peripheral blood mononuclear cells to kill red blood cells (RBC) from different species selectively. Cytotoxicity was induced by both mitogenic components of phytohemagglutinin-P (PHA), erythroagglutinating (E-PHA) and leukoagglutinating (L-PHA), and the non-mitogenic lectin wheat germ agglutinin (WGA). The target cells used in an overnight chromium release assay included human autologous RBC, human allogeneic RBC, and xenogeneic RBC from sheep or chickens. Although E-PHA induced cytotoxicity for all cell types, L-PHA caused human mononuclear cells to kill only xenogeneic RBC and, conversely, WGA induced killing of only human RBC. These differences allowed further investigation of possible control mechanisms.

The target cell specificity associated with lectin-induced cellular cytotoxicity did not correlate with lectin binding to, or agglutination of, the different red blood cell types. Furthermore, preincubation of RBC with lectins followed by washing did not result in cytotoxicity. However, when the mononuclear cells were preincubated with the lectins, the same cytotoxic specificity was observed as when lectins were present during the entire assay. These experiments suggest that the target cell specificity observed with lectin-induced cellular cytotoxicity is related to a prearmed lymphocyte which seeks out and kills the appropriate target cell.
Response of Human Lymphocytes to PHA and Con A, Dependent on and Regulated by THF, A Thymic Hormone
Authors: Rotter Varda , Schlesinger Menachem , Kalderon Rina , Trainin Nathan
Published: December 2022

The response of human lymphocytes to T lectins was shown to be dependent on and regulated by THF (thymus humoral factor). When human lymphocytes were stimulated with T lectins in the presence of dialyzed human plasma (DHP) which probably contains a low thymic hormone concentration, we observed a reduced response to PHA and Con A, compared to that observed in the presence of whole human plasma (WHP). This reduced reactivity to T lectins in the presence of DHP was restored by the addition of plasma dialyzates or THF. On the contrary, addition of THF to cultures in the presence of WHP caused a significant reduction in reactivity to T lectins, suggesting that enhancement or reduction of the response of human lymphocytes to T lectins by THF depends on the endogenous content of the thymic hormone in the plasma. From the present data it is suggested that the response of human cells to PHA behaves differently from that to Con A, as reflected in their dependence on THF. Although the response of human cells to Con A is increased by THF only in the presence of DHP, the response to PHA is also enhanced by THF in the presence of low WHP concentrations or even in the absence of WHP.
Specific Lysis of Human Colon Tumor Cells by Antibodies to CEA and Isoantigen A: Dependence on Rabbit Serum or Neuraminidase
Authors: Tompkins Wayne A. F. , Seth Pradeep B. , Yip Dick-Man , Palmer Jerry L. , Gee Sydney R. , Rawls William E.
Published: December 2022

The 51Cr cytotoxicity test was used to measure specific antibody reactions against carcinoembryonic antigen (CEA) and isoantigen A on the surface of human colon tumor cells. When human serum or guinea pig serum was used as a source of complement, no anti-CEA or anti-isoantigen A cytotoxicity was demonstrable despite binding of specific antibodies and activation of complement at least through the C3 component on the cell surface. In contrast, specific anti-CEA and anti-isoantigen A cytotoxicity was demonstrated when rabbit serum was used as a source of complement. Specific antibody-mediated cell lysis was also achieved with guinea pig complement if the cells were treated with neuraminidase before testing. These results support the concept that certain tumor cells have surface properties that render them resistant to immune lysis.
Mutant Lines of Guinea Pig L2C Leukemia
Authors: Konen T. G. , Green I. , Shevach E. M. , Kask A. M. , Schwartz B. D.
Published: December 2022

An alloantiserum was prepared in a strain 13 guinea pig against the GH line of the strain 2 guinea pig L2C leukemia. This serum contained antibodies to both IgM and Ia molecules. After absorption with normal spleen cells from a strain 2 guinea pig, this antiserum no longer reacted with strain 2 cells, but detected idiotypes on the IgM molecules of the L2C leukemia. These idiotypes were on the same IgM molecules detected by a xenogeneic sheep anti-L2C Fab µ antiserum. As assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the idiotype-bearing IgM molecules were synthesized by the cell, composed of normal sized µ and light chains, appeared on the cell surface as monomeric IgM, and were the only immunoglobulin molecules present on the cell. Although the alloantiserum potentially contained antibodies to unique Ia idiotypic determinants, none were found. Furthermore, the anti-IgM idiotype antisera did not react with any Ia-like molecules.
Induction of Antibody to the “y” Determinant of HBsAg in a Chimpanzee Carrier of HBsAg Subtype “adw”
Authors: Tabor E. , Gerety R. J. , Smallwood L. A. , Barker L. F.
Published: December 2022

Antibody to the y determinant of hepatitis B surface antigen (HBsAg) was induced in a chimpanzee chronically infected with hepatitis B virus and circulating HBsAg subtype adw. The chimpanzee was immunized with purified preparations of HBsAg subtypes adw and ayw. Six weeks after immunization, antibody to HBsAg (anti-HBs), specific for the y determinant, appeared. No change occurred in titers of HBsAg or antibody to hepatitis B core antigen (anti-HBc), and “e” antigen remained detectable. The circulating HBsAg subtype adw remained present despite the persistence of anti-y for greater than 8 months.
Genetic Control of B Cell Activation by Bacterial Lipopolysaccharide Is Mediated by Multiple Distinct Genes or Alleles
Authors: Glode L. Michael , Rosenstreich David L.
Published: December 2022

Eleven closely related C3H mouse strains were examined for differences in their DNA synthetic response to a highly discriminatory endotoxin, LPS Escherichia coli K-235. Strains of the C3H line were found to be either high, intermediate, or low responders to endotoxin. Three stains, C3H/St, C3H/Bi, and C3H/CR were high responders. Six strains, C3H/He, C3HfB/He, C3H/Hen, C3Heb/Fej, C3H/DiSn, and C3H/Avy were intermediate responders, suggesting that a mutation producing decreased LPS responsiveness occurred in the C3H/He(C3H/An) strain between 1931 and 1945. Two strains, C3H/HeJ and C3H/Bts were unresponsive due to a mutation that occurred between 1960 and 1968. Breeding experiments among high, intermediate, and low responding strains documented probable codominant genetic control by the genes or alleles in the C3H/HeN and C3H/HeJ strains. In the C3H/HeN × C3H/St cross, dominance of the C3H/St gene over the C3H/He gene was documented, whereas the C3H/HeJ × C3H/St cross indicated codominance of these two genes. These findings may represent either three alleles or three distinct genes in C3H mice. A second X-linked gene locus was documented in the CBA/N strain which also causes impaired B cell responsiveness to LPS E. coli K235 in serum-free conditions. The abnormal gene products in the CBA/N and C3H/HeJ strains exhibit complementarity since F1 female animals from the cross between these two unresponsive strains are responsive to LPS. We conclude that there is at least one distinct X-linked gene locus and either three additional autosomal genes or three possible alleles at one or more autosomal loci which determine the LPS sensitivity of murine B cells.
Mechanism of Mastocytoma-Mediated Suppression of Lymphocyte Reactivity
Authors: DeLustro Frank , Argyris Bertie F.
Published: December 2022

Previously we have shown that mitomycin-treated P-815 (H-2d) cells (P-815m) inhibit the in vitro response of C57BL/6 spleen cells to mitogens and DBA/2 alloantigens. The present data indicate that P-815m cells also inhibit the response of C57BL/6 spleen cells to AKR (H-2k)-stimulating cells and that the inhibition does not appear to be the result of crowding. Subpopulations of spleen cells obtained by density gradient centrifugation or after removal of glass-adherent cells are all sensitive to the inhibitory effects of P-815m. We also show that P-815m can inhibit the in vivo generation of graft-versus-host reactivity. Thus the inhibition due to P-815m cells appears to operate both in vitro and in vivo.

The experiments characterize the suppression of MLC reactivity by cell-free preparations of sonicated P-815 mastocytoma cells. The results suggest that the inhibition is not removed by ultracentrifugation, ultra-violet irradiation, or dialysis of the sonicate. P-815 sonicates subjected to chloroform extraction, heating at 56°C, 0.1 µ filtration, or treatment with anti-minute virus of mice serum are still inhibitory. This inhibition is resistant to RNAase treatment but destroyed by pronase. P-815 sonicate suppresses the response of C57BL/6 spleen cells to both DBA/2 and AKR-stimulating spleen. Inhibition of MLC reactivity is only seen if P-815 sonicate is added within the first 48 hr after the initiation of culture. These results indicate that the inhibition of MLC by sonicates of P-815 cells is due to a nondialyzable protein whose effects are not H-2 restricted and are acting on the early phase of sensitization.
Functional Heterogeneity Among the T-Derived Lymphocytes of the Mouse
Authors: Araneo Barbara A. , Marrack Philippa C. , Kappler John W.
Published: December 2022

Memory T cells were demonstrated in mice which were the precursors for cells causing sheep red blood cell-(SRBC) specific helper activity in the in vitro response of spleen cells to trinitrophenyl (TNP)-SRBC. These memory cells were distinguished from the effectors of helper activity generated during the primary response and were shown to belong to the T2 subpopulation of peripheral T cells on the basis of 1) their rapid response to a challenge with SRBC, 2) their long lifetime, and 3) their sensitivity to small in vivo doses of anti-thymocyte serum.
Structural and Genetic Studies on Chicken 7S Immunoglobulin Allotypes
Authors: Wakeland Edward K. , Benedict Albert A.
Published: December 2022

A method was developed for the detection of allotypes on intact, radiolabeled 7S immunoglobulin (Ig) which does not involve insolubilized alloantisera. Binding of dinitrophenylated anti-allotype (DAA) antiserum with radiolabeled alloantigen was demonstrated by the addition of rabbit anti-dinitrophenyl antiserum followed by precipitation of complexes with sheep anti-rabbit γ-globulin antiserum. Dinitrophenylation of alloantiserum up to 8.5 hr had no effect on antibody titer or avidity. Binding inhibition assays with the DAA method detected allotypes on 2 to 5 ng of 7S Ig. Four specificities were detected on 7S Ig heavy (H) chains by binding inhibition with the DAA assay. Specificities CS-1.1 and CS-1.2, located in the Fab fragment, were detected at 3- to 5-fold lower concentrations of reassociated H and light (L) chains than on the isolated H chains. On the basis of regression coefficients calculated from binding inhibition curves, the avidity of isolated H chains was significantly increased following reassociation with L chains, indicating that the conformation of CS-1.1 and CS-1.2 determinants depends in part on H and L chain interactions. In contrast, reassociation of H and L chains did not significantly increase H chain inhibitory activity or avidity for specificities CS-1.3 and CS-1.4.
Autoimmunity and Aging: The Age-Related Response of Mice of a Long-Lived Strain to Trinitrophenylated Syngeneic Mouse Red Blood Cells
Authors: Naor David , Bonavida Ben , Walford Roy L.
Published: December 2022

Mice of 1.5, 9, 22, and 31 to 32 months of age were injected with the thymus-dependent antigen, TNP-SRC, or the thymus-independent antigen, TNP-MRC. The anti-SRC and TNP immune responses to TNP-SRC were markedly reduced in older mice, whereas the anti-TNP response to the TNP-MRC showed no substantial decline. Young mice produced higher anti-TNP plaque-forming cell responses after injection of TNP-SRC than after TNP-MRC, whereas in older mice the reverse obtained. Old mice but not young mice displayed a high anti-SRC cross-reactive response after injection of TNP-MRC. The avidity of anti-TNP antibody of young mice immunized with TNP-SRC was higher than that following immunization with TNP-MRC, whereas the avidities of anti TNP antibodies from old mice injected with these two reagents were the same. Those individual mice which showed a poorly regulated immune response also displayed an autologous anti-MRC plaque-forming cell response after injection of either TNP-SRC or TNP-MRC. It is suggested that mechanisms mediated by suppressor T cells may be responsible for regulating the autoimmune response to modified self antigens, and that these are severely impaired in aged individuals.
Maintenance of Hyporesponsiveness to Antigen by a Distinct Subclass of T Lymphocytes
Authors: Weitzman S. , Shen F. W. , Cantor H.
Published: December 2022

Immunization with increasing doses of SRBC, in excess of 108, results in a progressive decline in the anti-SRBC PFC response. This hyporesponsive state is antigen specific and is reflected in a decrease of both T helper and B antibody-forming activity. We asked whether the apparent defect of T helper activity reflected a) an absence of αSRBC helper T cell activity, or b) the presence of SRBC-specific suppressor T cells within the hyporesponsive population. Our results indicate that at least a portion of hyporesponsiveness noted after antigen exposure to large doses of antigen can be ascribed to specific suppressor T cell activation.

Fractionation of the suppressive T cell population using Ly antiserum showed that specific suppressive activity was mediated by a subclass of T cells (Ly2+), distinct from that committed to express helper function (Ly1).
Cell-Mediated Immune Response in Vitro
Authors: Hirano Toshio , Nordin Albert A.
Published: December 2022

Spleen cells harvested from mixed leukocyte cultures (MLC) on day 2 or 3 suppress the development of CL from a fresh MLC across a cell-impermeable membrane, but day 4 MLC cells which have the maximum level of CL showed only a limited effect. Inhibition was observed only when suppressor cells were restimulated with the same H-2 type cells used during induction. However, the suppressive effect was not strain specific; that is, CBA-induced C57BL/6 spleen cells effectively inhibited the development of CL from DBA/2-induced C57BL/6 cells. In addition, DBA/2-induced C57BL/6 spleen cells effectively inhibited the development of CL from CBA cells. B10 spleen cells stimulated by B10.D2 cells gave rise to a suppressor cell population, indicating that H-2 differences alone can induce the response. The suppressive effect seemed to be exerted on an early phase of the response since no detectable inhibition was observed when suppressor cells were added 48 hr after culture initiation. The suppressive effect is not exerted on the accessory cell function but seems to inhibit DNA synthesis of the reacting cells in the MLC.
Co-Occurrence of Hepatitis B Surface Antigen of a Particular Subtype and Antibody to a Heterologous Subtypic Specificity in the Same Serum
Authors: Sasaki Takemichi , Ohkubo Yasuhito , Yamashita Yutaka , Imai Mitsunobu , Miyakawa Yuzo , Mayumi Makoto
Published: December 2022

In the routine screening of blood units for hepatitis B surface antigen (HB8Ag)2 and homologous antibody (anti-HB8) by counterimmunoelectrophoresis, we encountered two serum samples which showed precipitin lines both against anti-HB8 and HB8Ag. The HB8Ag in these serum samples were subtyped, and the specificity of co-occurring anti-HB8 was determined, in terms of the d, y, w, and r subspecificities found and described by Le Bouvier (1) and Bancroft et al. (2). HB8Ag/adr plus anti-w were found in one, and HB8Ag/adw plus anti-r in the other.

Both serum samples were obtained from apparently healthy individuals who voluntarily donated a blood unit at regional blood centers in Tochigi-ken and Osaka. They are hereafter referred to as Tog and Osk, respectively. Counterimmunoelectrophoresis was performed in 0.8% J-agar (Sankyo Junyaku Co., Tokyo) dissolved in Veronal buffer (pH 8.6, 0.05 µ). Three wells of 3 mm in diameter were cut in a line 5 mm apart (edge to edge) in an agar plate of 1.2-mm thickness.
Hereditary C5 Deficiency in Man
Authors: Breckenridge R. T. , Rosenfeld S. I. , Graff K. S. , Leddy J. P.
Published: December 2022

Platelet-rich-plasma from two hemostatically normal individuals, genetically lacking the fifth component of complement (C5), failed to exhibit normal platelet aggregation, or serotonin release, in the presence of zymosan. This abnormality was found to reside in the C5D plasma rather than in the platelets as demonstrated by the inability of the deficient plasma to activate zymosan for the aggregation of washed normal platelets. The defect could be corrected by the addition of normal plasma, normal serum, or highly purified human C5.

A plasma abnormality similar to that found in the C5D individuals was also noted in plasmas deficient in C3, C6, and C7; whereas C8 plasma D behaved normally. These data suggest that this platelet reaction requires late acting C components, perhaps as the complex, bound to the zymosan particles.
Ionophorous Activity and Murine B Lymphocyte Mitogens
Authors: Rosenstreich David L. , Blumenthal Robert
Published: December 2022

The relationship between ionophorous and B cell mitogen activity has been investigated. Most known ionophores were nonmitogenic for mouse spleen cells. In addition, when tested in a bilayer lipid membrane (BLM) apparatus, most types of B cell mitogens were nonionophorous.

However, excitability-inducing material (EIM), a high m.w. polymeric protein, which is a channel-forming ionophore, was a potent mitogen for mouse B lymphocytes. Similarly, keyhole limpet hemocyanin (KLH), a high m.w. polymeric protein, which is a B cell mitogen, is a channel-forming ionophore. The mitogenic activities of these two compounds were not due to contamination with endotoxin since they produced weak or absent responses in the limulus lysate clotting and rabbit pyrogenicity assays, and were also mitogenic for spleen cells of endotoxin-low responder C3H/HeJ mice.

Both the mitogenic and ionophorous activities of EIM and KLH were dependent on their polymeric structure since dissociation of these compounds into monomeric subunits markedly decreased both activities. However, heat denaturation destroyed their ionophorous ability but preserved their mitogenicity, thereby demonstrating that ionophorous activity was not essential for B cell activation. These data suggest that B cell mitogens do not necessarily act as primary ionophores. However, we propose that these molecules intercalate into the lipid portion of the cell membrane, and that this interaction initiates the process of B cell activation.
Immunosuppression of T Lymphocyte Function by Fractionated Serum from Tumor-Bearing Mice
Authors: McMaster Robert , Buhler Karen , Whitney Richard , Levy Julia G.
Published: December 2022

Sera from mice with transplanted 3-methylcholantrene-induced tumors have been shown previously to inhibit the function of normal lymphoid cells. When chromatographed on Sephadex G-150, the fraction eluting with immunoglobulin has been shown to inhibit the proliferative response of normal spleen cells to concanavalin A and to inhibit the in vitro antibody response to a T-dependent antigen, but has a lesser effect on the antibody response to a T-independent antigen. This paper deals with studies on the mode of action of the serum factor. The immunoglobulin containing fraction of serum from tumor-bearing mice inhibited the in vitro generation of both allogeneic and syngeneic cytotoxic lymphocytes. Time course studies demonstrate that the serum fraction inhibits the generation of antibody-producing and cytotoxic lymphocytes if added during the first 2 days of a 5-day culture. Serum fractions added after day 2 had no effect on the in vitro response. The serum factor appears to inhibit the generation of specific T cell function during the proliferative stage of development but has no effect on the differentiation stage which leads to either antibody-producing cells or cytotoxic lymphocytes.