bluuebunny/crossref_metadata_2025 huggingface dataset:
Selective effects of cyclosporin A on functional B cell subsets in the mouse.
Authors: Kunkl A , Klaus G G
Published: December 2022

Cyclosporin A, an immunosuppressive peptide of fungal origin, was believed to selectively affect T lymphocyte functions and to have minimal affects on B lymphocytes. This study shows that, in the mouse, T-dependent B cells and those responding to certain T-independent antigens (so-called TI-1 antigens) are indeed resistant to the drug. However, B cells responsive to other TI antigens (TI-2) are exquisitely sensitive. Thus, doses of the drug that completely abrogate responses to dinitrophenylated (DNP) Ficoll or dextran enhance the response to DNP-lipopolysaccharide and have minimal effects on the response to DNP-Brucella abortus. Virgin T helper cells are sensitive to the drug, whereas primed T cells are not. Cyclosporin A sensitivity therefore represents a novel marker of functional B cell subsets in the mouse and presumably points to fundamental physiologic differences between such subsets.
Radioimmunoassay for detecting antibodies against murine malarial parasite antigens: monoclonal antibodies recognizing Plasmodium yoelii antigens.
Authors: Kim K J , Taylor D W , Evans C B , Asofsky R
Published: December 2022

A solid-phase radioimmunoassay (SPRIA) in microtiter wells was established for detecting antibodies against Plasmodium yoelii Ag. The SPRIA was found 1) to require as little as 5 micrograms of crude parasite Ag per well, 2) to be able to detect 0.5 ng of monoclonal Ab, and 3) to be 10(4) times more sensitive than the indirect fluorescent Ab staining technique. In a modification of the above assay using intact RBC as an Ag, hyperimmune serum showed significant binding to the surface of erythrocytes of mice infected with P. yoelii parasites but not to RBC of normal mice. Hybridomas were prepared by fusing infected mouse spleen cells with P3-NS1/I-Ag4-1 myeloma cells. Using the SPRIA, hybrids secreting Ab against P. yoelii 17XL Ag were detected. All the hybrids detected by SPRIA secreted Ab binding to free parasites but not to the surface membrane Ag on the infected RBC. The number of positive hybrids produced by fusing spleen cells from mice given a double infection (17XNL and 17 XL) was not greater than that produced by freezing spleen cells from mice given a single infection (17 XL).
Human killer cells and natural killer cells: distinct subpopulations of Fc receptor-bearing lymphocytes.
Authors: Neville M E
Published: December 2022

Unstimulated human peripheral blood lymphocytes were depleted of K cells, which mediate antibody-dependent cellular cytotoxicity (ADCC) without removing NK cells, which mediate natural killing (NK). K cell depletion was achieved by buoyant centrifugation removal of lymphocytes that bound to glutaraldehyde-treated P815-AB cells at high lymphocyte-to-target ratios. Likewise, NK cells were removed with glutaraldehyde-treated K562 cells without removing K cells. Furthermore, both cytotoxic cell populations were observed directly in one agarose single-cell cytotoxic assay (ASCA) using P815-AB and K562 cells simultaneously as target cells. Moreover, the percentage of total cytotoxic cells was equal to the sum of the percentage of K and NK cells observed in separate ASCA. Collectively, these results indicate that K cells and NK cells are distinct subsets of FcR-bearing lymphocytes. One subset, K cells, has more avid Fc receptors (fcR) than NK cells and are 'activated' via thier FcR to kill antibody-coated target cells. The second subset, NK cells, have less avid FcR and are not 'activated' through their FcR to kill antibody-coated target cells.
Definition of a new T lymphocyte cell surface antigen, Ly 11.2.
Authors: Meruelo D , Paolino A , Flieger N , Offer M
Published: December 2022

The present communication defines a new cell surface antigen, Ly 11.2, which appears from its strain and tissue distribution to be distinct from all other previously defined normal or virally coded antigens or traits of the mouse. Lymphocytes of the T cell lineage, but not B cells or nonlymphoid cells, bear this marker. Although the locus coding for this antigen appears to be closely linked to the minor histocompatibility locus H-30 and to the locus coding for Ly 6.2, a chromosome assignment has not yet been made for my of these loci.
Characterization of the responding populations for the generation of proliferative response to syngeneic Meth A tumor in BALB/c mice: requirement of T and B cell collaboration.
Authors: Ting C C , Rodrigues D , Hong J X
Published: December 2022

High levels of primary proliferative response to a chemically induced sarcoma Meth A can be induced in syngeneic BALB/c spleen cells. Testing at the peak of proliferative response (2 days after sensitization in the mixed lymphocyte tumor cell culture), we found the responders to be resistant to anti-Thy 1.2 antibody lysis but susceptible to anti-Ia antibody lysis. When responders were subjected to various treatments before sensitization, it was found that removal of macrophages had no effect on the generation of proliferative response; high levels of proliferative response could be induced in enriched B cell preparations and in spleen cells from nude mice, but there was only a negligible amount of response in enriched T cell preparations. These findings indicate that the responders are primarily B lymphocytes. However, it was also found that the enriched B cell preparations usually gave only 50 to 75% of the response of whole spleen cells, whereas these B cells gave a 2- to 3-fold increase in the response to a B cell mitogen, LPS; this result indicate that collaboration from other types of lymphocytes was required for the generation of an optimal proliferative response to Meth A. Addition of 10% of T cells indeed produced a helper effect on this B cell response, and the maximal helper effect was seen for a mixture containing equal parts of T cells and B cells or for a slight T cell excess. These results indicate that the proliferative response to a syngeneic Meth A tumor is a macrophage-independent T-dependent B cell response.
Experimental allergic encephalomyelitis: activation of suppressor T lymphocytes by a modified sequence of the T effector determinant.
Authors: Hashim G A
Published: December 2022

Deletion of certain amino acid residues from the amino acid sequence of the encephalitogenic determinant for guinea pigs, H-Phe-Ser-Trp-Gly-Ala-Glu-Gly-Gln-Lys-OH, destroyed its ability to induce experimental allergic encephalomyelitis (EAE), a cell-mediated autoimmune disease of myelin. The administration of the modified determinant in the form of 4 repeating pentameric sequences, H-(Phe-Ser-Trp-Gln-Lys)4-Gly-OH, activated an antigen-specific T suppressor lymphocyte subset that rendered both presensitized donors and recipients of donor T lymphocytes unresponsive to an encephalitogenic challenge. Treatment of donors or recipients with cyclophosphamide before or after lymphocyte transfer, respectively, obliterated the ability of peptide S42-sensitized T lymphocytes to induce a state of unresponsiveness to an EAE-challenge. The results establish the existence of antigenic determinants for both immunoinduction and immunoregulation of EAE. The immunoregulatory determinant that activates antigen-specific and cyclophosphamide-sensitive suppressor T lymphocyte subset is sequestered within the disease-inducing or T effector determinant.
Macrophage-mediated tumor cell killing: regulation of expression of cytolytic activity by prostaglandin E.
Authors: Taffet S M , Russell S W
Published: December 2022

The expression of nonspecific macrophage-mediated tumoricidal activity in vitro is a transient phenomenon. The work reported here shows that the loss of tumoricidal activity by mouse macrophages is attributable, at least in part, to the effects of prostaglandin E (PGE). The amount of PGE needed to inhibit the expression of cytolytic activity (2 x 10(-9) M) was readily synthesized and secreted by the same population of resident peritoneal macrophages that had been induced to kill tumor cells by exposure to 100 ng/ml bacterial lipopolysaccharide (LPS). Inhibitory concentrations of PGE were present in supernatants within the 1st hr after macrophages were exposed to LPS. In spite of this fact, cytolytic activity still developed, and 12 to 16 hr were required for the PGE to have its full inhibitory effect. Thus, PGE did not act by blocking the development of cytolytic activity. Treatment of LPS-stimulated macrophages with 10(-6) M indomethacin (or other cyclooxygenase inhibitors) prevented both PGE synthesis and the shut-off of cytolytic activity. The latter effect could be reversed by adding PGE2 to the cultures at a concentration of 10(-8) M. Lastly, PGE inhibited cytolytic activity mediated by indomethacin-treated macrophages only when activator was present: pulsing with PGE before the addition of LPS had a negligible inhibitory effect. These findings may help explain why intratumoral macrophages often lack nonspecific cytolytic capabilities.
Antigen-specific human T cell factors. II. T cell suppressor factor: biologic properties.
Authors: Uytdehaag F , Heijnen C J , Pot K H , Ballieux R E
Published: December 2022

The in vitro induction of an ovalbumin-specific human T cell suppressor factor is described (TsF120-OA). The antigen-specific suppressive component can be purified by affinity chromatography from supernatants derived from Marbrook-Diener type cultures of peripheral blood T cells stimulated with a high dose of ovalbumin. TsF120-OA suppresses the antigen-induced PFC formation of human blood B cells in vitro in an antigen-specific way. The target of TsF120-OA activity is shown to be the T helper cell. No genetic restriction in the action of the factor is observed.
I-region-restricted long-term cultured T cells with helper function for thymic cytotoxic precursor cells.
Authors: Guimezanes A , Schmitt-Verhulst A M
Published: December 2022

T cells with specific helper function (TM) for the differentiation of thymic cytotoxic T cell precursors (pCTL) into anti-self-TNP effector cells could be enriched in vitro by multiple stimulations with TNP-syngeneic cells after in vivo sensitization with the same hapten. H-2K/D-restricted cytotoxic effector cells (CTL), initially present in such cultures, were often lost after 4 to 6 stimulations without affecting the maintenance of specific TM. I-region homology between responding and hapten-treated stimulating cells was necessary both to induce strong proliferative responses and to maintain the propagation of cells with TH function. The analysis of the interaction between the lines of TH and pCTL indicated that H-2 homology was required between thymocyte responding cells, helper lines and hapten-treated stimulating cells in order to obtain anti-hapten self CTL, but Ig-1 allotype homology was not required.
Binding of C1q and complement activation by vascular endothelium.
Authors: Linder E
Published: December 2022

Exposure of intermediate filaments of cultured cells to serum leads to antibody-independent binding of complement (C) component C1q, C4, and C3. The apparent C-activating property of cytoskeletal intermediate filaments was examined on a tissue level in The present study by using frozen sections of human kidney and term placentas as targets. The assay used fresh human serum or isolated C1q as sources of C and the bound C components were demonstrated by fluorescent conjugates against C1q, C4, and C3. A consistent finding was that the endothelium of vessel walls had the capacity to bind both C1q and C components C4 and C3. Less intense binding of C to vascular smooth muscle was noted. Binding of C1q was very rapid, taking place within 10 sec. Subsequently, accumulation of C4 and C3 was seen. Subsequently, accumulation of C4 and C3 was seen. Upon prolonged incubation, a decrease in demonstrable C1q and C4 was observed. Unfixed vascular endothelium bound both isolated C1q and C1q of EDTA- or enzyme inhibitor-treated serum. However, if unfixed, the binding structures appeared to be destroyed in the process of C activation. They were also readily destroyed by trypsin. The described observations seem to suggest a possible mechanism whereby C components may become deposited in vessel walls independently of antibodies.
Giardiasis in mice: studies on the characteristics of chronic infection in C3h/He mice.
Authors: Underdown B J , Roberts-Thomson I C , Anders R F , Mitchell G F
Published: December 2022

When C3H/He mice are dosed orally with fecal cysts or trophozoites of the intestinal parasite, Giardia muris, they become chronically infected, whereas BALB/c (or DBA/2) mice rapidly eliminate their infection. The ability to eliminate a primary infection or resist a challenge infection was not related to levels of serum IgA antibody directed against a crude extract of trophozoites. The possibility that the development of host-protective immunity may either be delayed or absent in C3H/He mice was investigated by 2 types of experiment: 1) BALB/c and C3H/He mice were drug-cured at various times after a primary infection and assessed for their ability to resist reinfection, and 2) the ability of infected female mice to protect their suckling young was assessed. Under these experimental conditions, no significant difference in the kinetics of development of resistance to a challenge infection was noted. Thus, mice of both strains were capable of resisting a challenge infection after drug-cured and infected mothers were able to protect their suckling young from a challenge infection. Drug-cured long-term infected mice were resistant to a challenge of either cysts or trophozoites obtained from either 1-wk infected nude mice or long-term infected C2H/He mice. Thus, it is unlikely that the explanation for chronic infection oif C3H/He mice is that variants or a modified population of G. muris trophozoites arise to which C3H/He mice have a defective response. The possibility that G. muris is able to suppress host-protective responses in chronically-infected C3H/He mice but not in BALB/c mice is discussed.
IgD-secreting murine plasmacytomas: identification and partial characterization of two IgD myeloma proteins.
Authors: Finkelman F D , Kessler S W , Mushinski J F , Potter M
Published: December 2022

We have used an immunofluorescence inhibition assay to identify 2 BALB/c plasmacytomas, TEPC-1017 and TEPC-1033, that secrete large quantitites of IgD. Both TEPC-1017 and TEPC-1033 myeloma proteins bound to anti-kappa as well as hybridoma and heterologous anti-delta antibodies, but not to anti-mu, gamma, alpha, or lambda antibodies. Both myeloma proteins were purified by (NH4)2SO4 precipitation, ion exchange chromatography, gel filtration, and Staphylococcus aureus Protein A absorption. These IgD kappa myeloma proteins were used to prepare affinity purified rabbit antibodies to delta-chain and the TEPC-1017 and TEPC-1033 idiotypes. Native TEPC-1017 and TEPC-1033 both had mobilities between those of mouse IgA kappa dimers and trimers when analyzed by polyacrylamide gradient gel electrophoresis. Both IgD myeloma proteins broke down under mild reducing conditions into subunits with electrophoretic mobilities slightly slower than those of an IgA kappa monomer. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of reduced TEPC-1017 and TEPC-1033 demonstrated kappa-chains and heavy chains that co-migrated with alpha chain. These data suggested that secreted IgD contains 2 delta 2 kappa 2 subunits that are linked by an easily reducible disulfide bond. The kappa-chains of IgD secreted by TEPC-1017 and TEPC-1033 have apparent m.w. of approximately 63,000 daltons, whereas the apparent m.w. of intracytoplasmic delta-chain, intracytoplasmic delta-chain synthesized in the presence of tunicamycin, and the cellfree translation product of TEPC-1017 delta-chain mRNA are 54,000, 43,000, and 44,000 daltons, respectively. This is compatible with the interpretation that the delta-chain peptide has a leader sequence and is N-glycosylated during or shortly after peptide synthesis and is glycosylated further shortly before IgD secretion.
Evidence for direct control of an in vitro plaque-forming cell response by quantitative properties of intact, fluid, haptenated liposomes: a potential model system for antigen presentation by macrophages.
Authors: Humphries G M
Published: December 2022

Stimulation of the primary in vitro plaque-forming cell (PFC)2 response to fluid, haptenated liposomes by spleen cells depleted of a Sephadex-adherent population has been made possible by the addition of an uncharacterized cell-derived soluble factor(s). In its absence, PFC responses by such cells are greatly reduced or absent. The factor(s) is present in the supernatant from Concanavalin A-stimulated spleen cells. This phenomenon has permitted a comparison of the behavior of cultures with and without adherent cells, with a view to determining the relative likelihood of liposomes or adherent cells functioning as antigen presenters in this system. Two modes of control by quantitative properties of liposomes have been studied. PFC stimulation is controlled, in a biphasic manner, by varying either liposomal epitope density at constant liposome concentration or liposome concentration at fixed epitope density. Overall hapten concentration in the cultures is only of significance as a consequence of epitope density and liposome concentration; it does not, itself, control the response. Whole spleen cell cultures and cultures depleted of Sephadex-adherent cells but supplemented with soluble factor(s) exhibit the same biphasic response profiles when these quantitative properties of liposomes are varied. The results argue against the role of Sephadex-adherent cells as antigen presenters in whole spleen cell cultures. A model in which antigen presentation is accomplished by intact liposomes, in both whole spleen and adherent cell-depleted cultures, is consistent with the data.
Reactions between polyribonucleotides and low density lipoproteins in sera of patients with progressive systemic sclerosis.
Authors: Garcia I , Wolbers R , Tan E M
Published: December 2022

A high percentage (58%) of sera from patients with progressive systemic sclerosis reacted with synthetic polyribonucleotides to give precipitin lines. This was demonstrated by counterimmunoelectrophoresis. The synthetic polyribonucleotides included polyuridylic acid (poly U), polyadenylic acid (poly A), and co-polymers of poly A-poly U. The reactive factor in serum was not immunoglobulin but was associated with low density lipoprotein.
Helper cells in murine thymus for the generation of primary cytolytic T lymphocyte (CTL) responses in vitro.
Authors: Wu S , Bach F H
Published: December 2022

Thymocytes used as responding cells in a mixed leukocyte culture with gamma-irradiated splenic stimulating cells generated highly significant proliferative and cytolytic responses when responding and stimulating cells differed by the entire H-2 complex or at H-2K plus H-21 regions. On the other hand, when the difference was only an H-2K region, very little, if any, proliferative response was detectable and no cytolytic response was found. Because it has been claimed by others that thymocytes do not include cells needed to provide the help required for the generation of a cytolytic response and yet we have found a highly significant response in thymocytes, we have analyzed, in 2 systems, the possible role of back-response by stimulating spleen cells in the generation of cytolytic activity by responding thymocytes. In the first system, spleen cells were depleted of T lymphocytes by treatment with anti-Thy-1.2 antiserum plus complement. In the second system, F1 hybrid mouse spleen cells were used to stimulate parental responding thymocytes. In both cases, no back-response as measured by cell proliferation in mixed leukocyte culture was detected, while both types of spleen cells still stimulate significant cell proliferation and cytolytic activity from responding thymocytes, as well as from responding spleen cells. Furthermore, it was found that in the presence of stimulating spleen cells (either T lymphocyte-depleted or not) that differ from the responding thymocytes at the entire H-2 complex, thymocytes were capable of generating a significant and specific cytolytic response to H-2K region different stimulating spleen cells (either T lymphocyte-depleted or not). These results are compatible with the suggestion that there are sufficient numbers of helper cells in the unfractionated thymocyte population for the generation of primary cytolytic T lymphocyte responses in vitro.
Sex-related differences in the rapid production of cytotoxic spleen cells active against uninfected myofibers during Coxsackievirus B-3 infection.
Authors: Huber S A , Job L P , Auld K R , Woodruff J F
Published: December 2022

Spleen cells from BALB/c mice immunized with a cardiotropic strain of Coxsackie B-3 virus were more cytolytic to uninfected myofibers than were spleen cells from mice immune to a noncardiotropic strain of the virus. Spleen cells immune to either virus were equally cytolytic to endothelial cells. Cytotoxicity was greater in female mice than in males. Analysis of individual reactivities showed that the male response was heterogeneous with only half of the animals developing cytolytic activity. All females responded. Early (day 3) cytolytic cells in both male and female mice appear to be natural killer (NK) cells, since they are not sensitive to anti-thy 1.2 or anti-Ig serum and complement, and lyse allogeneic (CBA) as well as syngeneic (BALB/c) targets. Later (days 4 to 6), the cytolytic cells in males become sensitive to anti-thy 1.2 serum and are restricted to lysis of syngeneic targets, while the cytolytic cells in females maintain the characteristics of NK cells.
Alloreactive cloned T cell lines. III. Accessory cell requirements for the growth of cloned cytolytic T lymphocytes.
Authors: Lutz C T , Glasebrook A L , Fitch F W
Published: December 2022

We examined the role of accessory cells that cooperated with the cloned cytolytic T cell line, L3. Accessory cells and T cell growth factors (TCGF) acted synergistically to stimulate cloned L3 CTL to grow and produce cytolytic activity. Accessory cells were non-T, radioresistant, plastic adherent M phi-like cells that interacted with cloned L3 CTL via soluble factors. Peritoneal M phi, at low density, also cooperated with L3 cells but at high density, inhibited L3 cells. Both accessory and inhibitory M phi acted upon L3 CTL without the intervention of other T lymphocytes or nonadherent cells. Other published reports indicate that M phi may promote cytolytic activity indirectly, amplifying TCGF production by helper T lymphocytes. This report indicates that M phi may also promote cytolytic activity directly, amplifying the response of CTL to TCGF.
Elicitation of anti-H-2 cytotoxic T lymphocytes with antigen-modified H-2 negative stimulator cells.
Authors: Hale A H , Ruebush M J , Harris D T , McGee M P
Published: December 2022

By a newly developed technique, we have modified H-2k-negative cells in such a way as to render them capable of elicitation of anti-H-2 CTL. Purified H-2Kk and/or partially purified Iak glycoproteins were incorporated into lipid vesicles that contained the hemagglutinin-neuraminidase (HN) glycoprotein and the fusion (F) glycoprotein of Sendai virus. These lipid vesicles were incubated with H-2-negative tumor cells (R1-) in a manner that resulted in rendering these cells capable of eliciting anti-H-2Kk CTL. The presence of Iak on the surface of the modified stimulator cell had a significant effect on the proportion of anti-H-2Kk CTL possessing a particular Lyt phenotype. Primary effector cells elicited by an H-2Kk-Iak difference were primarily Lyt 1-, 2+, whereas primary anti-H-2Kk CTL elicited by an H-2Kk difference alone were predominantly Lyt 1+, 2+.
Distribution of t lymphocyte subsets in the human bone marrow and thymus: an analysis with monoclonal antibodies.
Authors: Janossy G , Tidman N , Papageorgiou E S , Kung P C , Goldstein G
Published: December 2022

A panel of reagents (OKT1, 3, 4, 6, 8, and 11) detects differentiation antigens expressed exclusively on HuTLA+ T lymphoid cells but absent on bone marrow precursors, such as terminal deoxynucleotidyl transferase- (TdT) positive cells, immature myeloblasts, and other myeloid/erythroid cell types. In the bone marrow no transitional forms could be detected between TdT+ and T cells. The BM T cells showed mostly the suppressor/cytotoxic phenotype (OKT8+) with only a few T cells of inducer type (OKT4+). Thymocyte heterogeneity was also analyzed directly in tissue sections and in double labeling assays in combination with TdT staining, a marker for cortical thymocytes. Many large thymic blasts (in fetal and infant thymus) showed reactivity with OKT11--a pan-T reagent, but had only weak or negligible activity with the other antibodies. Cortical thymocytes reacted strongly with OKT11, 6, 4, and 8, whereas medullary cells reacted with OKT11, 3, 4 (majority) and 8 (minority). Thus, the reactivity with these antibodies is generated in the thymus at various stages of differentiation. In contrast, OKT10 (an anti-"precursor cell" reagent) reacted not only with thymocytes but also with TdT+ BM precursors, myeloblasts, and BM B lymphocytes although it was unreactive with mature peripheral lymphoid and maturing myelo/erythroid cells.
Functional and partial chemical characterization of the carbohydrate moieties of the IgE receptor on rat basophilic leukemia cells and rat mast cells.
Authors: Pecoud A R , Ruddy S , Conrad D H
Published: December 2022

The role of the carbohydrate portion of the receptor for IgE in the interaction with IgE was investigated. Membrane carbohydrates of rat basophilic leukemia (RBL) cells and rat mast cells (RMC) were labeled by treating the cells with galactose oxidase followed by [3H]-NaBH4. IgE receptors were separated from detergent solubilized membranes and examined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Pretreatment with neuraminidase markedly increased the incorporation of 3H into both the total membrane extract and into the IgE receptors. SDS-PAGE analysis demonstrated the presence of galactose in all IgE-binding components of 2 RBL cell lines and the presence of sialic acid on the major IgE-binding component. Prior saturation of the cells with IgE did not prevent the carbohydrate labeling of the receptor, although it did block the labeling of its protein part, indicating that carbohydrates are not located in the binding site. Removal of terminal sialic acid residues with neuraminidase increased the affinity of the receptor for IgE without appreciably affecting the number of receptors per cell. In order to more drastically modify the receptor carbohydrates, RBL cells were grown in the presence of Tunicamycin (TN). TN was shown to markedly inhibit the incorporation of [3H]-glucosamine into the receptor. RBL cells grown in the presence of TN expressed fewer receptors at the cell surface, as judged both by ligand binding studies and external labeling procedures. These data cumulatively suggest that the carbohydrate moieties of the receptor are not directly located in the binding site of the IgE receptor; however, the TN studies suggest that receptor carbohydrate may play a role in transport of the receptor to the plasma membrane or in its orientation thereafter.
Immunochemical evidence in two haplotypes for at least three D region-encoded molecules, D, L, and R.
Authors: Hansen T H , Ozato K , Melino M R , Coligan J E , Kindt T J , Jandinski J J , Sachs D H
Published: December 2022

Sequential immunoprecipitation analyses have defined a new transplantation antigen, designated R, which in addition to the 2 previously isolated D and L molecules is encoded in the D region. All 3 of these gene products are 45,000 m.w. glycoprotein, and each bears a unique combination of specificities as recognized by monoclonal and/or conventional anti-H-2 sera. Three D region products, D, L, and R, have now been isolated from soluble antigens of both the H-2d and H-2q haplotypes. The resulted reported here also indicate that the loss-mutant BALB/c-H-2dm2 fails to express both Ld and Rd antigens. Further chemical comparisons of the primary structure of R molecules with D and L molecules will now be necessary to determine whether R antigens are the products of a unique gene, as opposed to a glycosylation or conformational variant of D and/or L molecules. In either case, the discovery of 3 D region-encoded gene products in certain haplotypes raises new questions about the evolution and regulation of expression of H-2 loci.
The histocompatibility-2 system in wild mice. XII. An immunochemical analysis of the anti-Ia antibodies in antisera produced against 13 B10.W lines.
Authors: Wakeland E K , Klein J
Published: December 2022

The I-region products of 13 congenic B10.W lines carrying wild-derived H-2 haplotypes were analyzed with anti-B10.W alloantisera. The alloantisera were tested for immunoprecipitating activity with radiolabeled glycoproteins of the immunizing donors. Eleven of the 13 alloantisera immunoprecipitated glycoproteins with SDS-PAGE mobilities consistent with m.w. of 45,000 35,000, and 26,000. The 35,000 and 26,000 products were identified as the Ia antigens of the B10.W lines. The specificity of the anti-Ia antibodies in each anti-B10.W alloantiserum was assessed by panel testing and absorption analysis with a panel of 26 glycoprotein fractions that were prepared from 11 inbred lines and 15 B10.W lines. Four new Ia antigens that are only present in the B10.W lines were detected in this analysis. The I-region products of 8 of the B10.W lines were serologically indistinguishable from those found among laboratory inbred strains. The implications of these findings for I-region polymorphisms are discussed.
A fluorescent assay for complement activation.
Authors: Caporale L H , Gaber S S , Kell W , Götze O
Published: December 2022

We report here a rapid assay for the complement enzymes CVFBb, C4b2a and Cls. This assay involves the use of a peptide substrate that releases a fluorescent coumarin derivative (AMC) upon cleavage by the convertase. The substrate, BocLeuGlyArgAMC, was chosen because its sequence is similar to the carboxyl terminus of C3a, and identical to that of C5a. The Km of this substrate are about 125 microM for the C3/5 convertase CVFBb, 169 microM for C4b2a, and 140 microM for C1s.
T cell lines active in the delayed-type hypersensitivity reaction (DTH).
Authors: Weiss S , Dennert G
Published: December 2022

Two allospecific uncloned T cell lines, C.C3.11.75 (H-2d anti-H-2k) and B6-C.7.76 (H-2b anti-H-2d), were assayed for their ability to elicit a DTH reaction in vivo. It was shown that both cell lines, C.C3.11.75 being cytolytic and B6-C.7.76 being noncytolytic, evoke a strong DTH reaction. The reaction was in tempo and histology typical for a DTH reaction and could not be distinguished from a DTH reaction elicited by in vivo generated T cells active in a GVH reaction. Both cell lines recognize a private specificity encoded in the H-2 IA subregion. Cell proliferation of the responder cells did not appear to be critical in the reaction nor are T cells from the host recruited during the reaction. The DTH reaction can be assayed in allogeneic hosts demonstrating a complete lack of H-2 restriction in the reaction even when presumably antigen presenting macrophages are removed from the stimulator cell population. Furthermore, tumor cells expressing IA antigens could be employed to cause the reaction in both syngeneic and allogeneic recipients. Cloned sublines of one of the lines are also active in the reaction.
The human LT serum. X. The initial form released by T-enriched lymphocytes is 150,000 m.w., associated with small nonlytic components, and can dissociate into the smaller alpha, beta, and gamma m.w. classes.
Authors: Harris P C , Yamamoto R S , Crane J , Granger G A
Published: December 2022

The present studies examine the various lymphotoxin (LT) forms released in vitro by phytohemagglutinin- (PHA) activated T-enriched (Te) human peripheral blood lymphocytes. It is clear that Te cells rapidly released (24 to 48 hr) these molecules in vitro. The 1st cell-lytic form detected in these supernatants is a 140-160,000 m.w. molecule(s) termed precursor alpha heavy (P alpha H). This form does not express alpha-LT antigenic determinants but is neutralized by antisera from animals injected with serum-free PHA-activated unseparated lymphocyte supernatants (anti-WS). The P alpha H is converted into alpha H, which expresses alpha determinants, by passage through molecular sieving columns or by treatment with low levels of Nonidet P-40 or urea. These treatments dissociate a small nontoxic 10-20,000 m.w. molecule(s), termed precursor factor (Pf), which masks the alpha-LT determinant on the P alpha H molecule. The dissociation of Pf is reversible, since alpha H from the molecular sieving columns will reassociate with the Pf. The alpha H LT class can further dissociate into the smaller alpha, beta, and gamma LT forms upon chromatography on a molecular sieving column, and a certain small percentage of the alpha H forms appear capable of associating to form the high m.w. complex (Cx) LT class. These findings suggest P alpha H may represent an intermediate that requires additional processing in order to proceed down 1 of 2 pathways: a) formation of complexes that are highly cell-lytic, or b) degradation by dissociation into the smaller weakly cell-lytic molecules identified as LT forms.
Immunologic control of the ascites form of murine adenocarcinoma 755. III. Efficacy of serum therapy is controlled by a single genetic locus.
Authors: Roloson G J , Haagensen D E , Chambers C A , Bolognesi D P
Published: December 2022

We have previously described a passive serum therapy system in which potent protection against challenge of syngeneic mice with large doses (10(4) X LD100) of AD755a tumor cells can be accomplished by administration of as little as 2 to 5 microliters/mouse of hyperimmune anti-tumor cell serum. The present results demonstrate that the efficacy of the serum protection effect is strain-dependent and is under the control of a single genetic locus, denoted ADP-1. Through a series of crosses between highly protected C57BL/6J and nonprotected BALB/cJ mice, the ADP-locus has been mapped to chromosome 2 [linkage group V], approximately 23 centimorgans toward te centromere from the locus controlling agouti coat color. Thus, the protection locus is not linked to either the H-2 region or to previously described loci regulating the replication and disease induction of type-C viruses closely related to the virus associated with the AD775a tumor. The possible functions of the ADP-1 locus are discussed and further aspects of this experimental model are described in the accompanying papers.
Cross-reactions between mouse Ia and human HLA-D/DR antigens analyzed with monoclonal alloantibodies.
Authors: Pierres M , Rebouah J P , Kourilsky F M , Dosseto M , Mercier P , Mawas C , Malissen B
Published: December 2022

Two mouse monoclonal anti-I-E/Ck alloantibodies (H7-8.26 and H10-81.10) directed against 2 distinct determinants of the specificity Ia-7 and 1 anti-I-Ak alloantibody (H8-15.9) directed against a public determinant common to the I-A subregion products of the H-2k, H-2b, H-2d, H-2q, and H-2ja haplotypes identified cross-reactive determinants on lymphoid cells from various mammalian species, including rat, dog, pig, cow, hamster, and guinea pig. In man, these antibodies detected nonpolymorphic determinants of DR antigens on B cell-enriched peripheral blood lymphocytes from 50 unrelated individuals. These cross-reactive DR determinants were also detected on lymphoblastoid B cell lines, on PHA-activated peripheral T lymphocytes, and on allospecific cytolytic T cell clones, but not on various DR-negative human T leukemia cell lines. Two chains of 29,000 and 35,000 daltons m.w., corresponding to DR antigens, were precipitated by H7-8.26 and H8-15.9 antibodies from radiolabeled membrane extracts of Raji cells. Competitive binding experiments indicated that the 3 mouse anti-Iak antibodies identified 3 distinct cross-reactive determinants on human cells. The results indicate that: a) The cross-reactivity described between mouse I-E/C gene products (Ia-7) and human DR antigen(s) involves, in fact, several distinct and topologically distant determinants. b) At least 1 determinant cross-reacting with DR can be identified on I-Ak gene products. c) The intriguing genetic problem of mouse MHC allotypic determinant(s) being nonpolymorphic in man cannot be simply explained by the deletion of an I-E alpha chain in some strains of mice.
Activation of autologous reactive helper T lymphocytes for differentiation of human B lymphocytes.
Authors: Shin H S , Wang C Y , Choi Y S
Published: December 2022

We present experimental evidence that autologous reactive T cells are triggered to proliferate by HLA-D or Ia-like antigens on pokeweed mitogen-pulsed non-T cells and that a subpopulation of these proliferating cells in turn helps PWM-primed B cells to differentiate into immunoglobulin-secreting cells. These helper T cells displayed a proliferation kinetics different from that of other autologous reactive T cells in peaking on the second day of culture and were relatively resistant to irradiation or treatment with mitomycin C. Since heterologous anti-Ia antibody (F(ab')2 reagent) inhibits proliferation of helper T cells and the generation of plaque-forming cells, Ia antigen is an essential component in PWM-stimulated B cell differentiation. Anti-Ia reagent was shown to interfere with B cell differentiation in 2 ways. First, it abrogates the ability of PWM-primed non-T cells to activate helper T cells. Second, it directly affects a differentiation of B cells into Ig-secreting cells even in the presence of the activated helper T cells.
Experimental allergic encephalomyelitis in Lewis rats: inhibition by bacterial lipopolysaccharides and acquired resistance to reinduction by challenge with myelin basic protein.
Authors: Raziuddin S , Kibler R F , Morrison D C
Published: December 2022

In 2-mo-old Lewis rats immunized with bacterial lipopolysaccharides (LPS) precomplexed to guinea pig myelin basic protein (BP), the clinical and histologic manifestations of experimental allergic encephalomyelitis (EAE) were diminished compared with BP-treated controls. Similarly, in animals immunized with BP and challenged with BP-LPS at the same time or as long as 5 days after, the immunization with BP also inhibited the disease. That this capacity to reduce the incidence of BP-induced EAE is a unique property of LPS was suggested by the fact that other negatively charged molecules, such as DNA, RNA, and dextran sulphate, were not effective in inhibiting the clinical signs of EAE. After recovery from EAE induced by BP, some animals develop a recurrence of the disease if challenged with BP at appropriate intervals. However, after recovery from mild EAE induced by BP-LPS and after challenges with EAE-initiating BP antigens, secondary EAE was inhibited significantly.
Genetics of a non-H-2 antigen that stimulates "unrestricted" helper T cells and MLR.
Authors: Coutinho A , Augustin A A , Forni L , Fischer-Lindahl K
Published: December 2022

The inheritance of antigens expressed by C3H/Tif B cells that stimulate MHC-unrestricted helper T cells from C3H/HeJ was investigated. F1 hybrids between C3H/HeJ and C3H/Tif and 39 C3H/HeJ X F1 backcross mice were characterized as to the ability of their spleen cells to stimulate a proliferative C3H/HeJ T helper cell response and to respond to helper cell activity by the development of polyclonal plaque-forming cell responses. Backcross progeny wee also typed for the following markers segregating in this cross: 1) Responsiveness to the B cell mitogen lipopolysaccharide (LPS); 2) LyM-1 allotype; 3) antigen(s) stimulating a primary non-H-2 MLR between these strains, previously ascribed to Mls locus differences, 4) expression of target antigens for cytotoxic T cells raised in the same strain combination. The antigen(s) recognized by helper cells and those stimulating primary MLR are controlled by autosomal gene(s) and segregate as a single trait. These antigens, however, are not encoded in genes linked to either the Lps or the Mls loci, and are not recognized by cytotoxic T cells raised in the same strain combination.
Antigen-specific helper factor reacts with antibodies to human beta 2 microglobulin.
Authors: Lamb J R , Zanders E D , Sanderson A R , Ward P J , Feldmann M , Kontiainen S , Lehner T , Woody J N
Published: December 2022

Antigen-specific helper factor was induced in vitro from lymphoid cells of monkeys and mice by using an antigen derived from Streptococcus mutans. Helper activity was removed from supernatants of monkey cells by affinity chromatography on Sepharose 4B insolubilized antibodies specific for human beta 2-microglobulin (H beta 2M) prepared in chicken, rabbit and rat, and an insolubilized monoclonal mouse anti-H beta 2M antibody-bound monkey helper factor activity. However, guinea pig antibody to human beta 2M was inactive. In parallel studies, the pattern of absorption of mouse helper factor (HF) was different from that of the monkey in that insolubilized guinea pig anti-H beta 2M bound helper factor, whereas rabbit and monoclonal anti-H beta 2M failed to do so. Although these findings were not compatible with an intact beta 2M chain being present in helper factor, they may imply a cross-reactivity of beta 2M with a "constant region" of helper factor that may share common sequences with beta 2M. This may suggest that factor genes have evolved from the same ancestral genes as beta 2M.
Cellular interactions affecting the maturation of murine B lymphocyte precursors in vitro.
Authors: Kincade P W , Lee G , Paige C J , Scheid M P
Published: December 2022

Clonable B cells arose in liquid cultures of slg+ cell-depleted bone marrow suspensions, and a majority of these derived from rapidly sedimenting (presumably large) precursors. Such immature (pre-B) cells were not demonstrable in adult lymph nodes. The development of functional B cells in vitro depended on adherent cells present in adult bone marrow or peritoneal exudates and in the presence of optimal numbers of these accessory cells; not FCS or 2-ME or LPS were essential for B cell maturation. In contrast, the ability of fetal liver cell suspensions to generate B cells in culture was not affected by the removal of adherent cells or addition of macrophages, and LPS consistently reduced numbers of colony-forming B cells recovered. A type of nonadherent cell present in the bone marrow of adult CBA/N mice dramatically enhanced B cell maturation in normal CBA/H-T6T6 fetal liver cell cultures. These results indicate that 2 types of accessory cells may augment transition of pre-B cells to functional B cells in culture. One is adherent and can be replaced by macrophages, whereas the other is nonadherent and is present in limiting numbers in embryos. When fetal liver cultures were incubated with optimal numbers of adult accessory cells, the rate of emergence of B cells was dependent on the gestational age of the donor embryo. This suggests that B cell formation in utero may be limited by the availability of B cell precursors as well as nonadherent accessory cells. The possibility exists that fetal pre-B cells typify intermediates in the B lineage which are infrequent in adult marrow. Alternatively, B cells may be produced in adult life through processes that do not precisely recapitulate embryonic events.
Differential anti-inflammatory effects of LPS in susceptible and resistant mouse strains.
Authors: Verghese M W , Snyderman R
Published: December 2022

Most mouse strains are highly susceptible to endotoxin (LPS) lethality and are responsive to LPS stimulation in vitro (e.g., B cell mitogenesis, macrophage activation). They are, however, capable of mounting only a small inflammatory response to LPS when it is injected i.p. The present study demonstrates that LPS is in fact a potent, anti-inflammatory agent in all of the five normally LPS susceptible strains tested. LPS was also anti-inflammatory in F1 hybrid mice from susceptible (C3HeB/FeJ) x-resistant (C3H/HeJ) parents. Anti-inflammatory effects of LPS in susceptible strains were achieved by either i.v. or i.p. treatment and were observed toward a variety of phlogistic stimuli including mitogens, C-activating substances, and nonspecific irritants. The most dramatic inhibitory effect of LPS was directed toward the accumulation of inflammatory macrophages. Kinetic studies indicated that the anti-inflammatory effect of a single dose of LPS persisted for at least 72 hr but was maximal when LPS was given simultaneously with the inflammatory stimulus. In contrast to normal mice, two mutant, LPS resistant strains (C3H/HeJ and C57BL10/ScCR) responded to increasing doses of LPS i.p. with a progressively increasing influx of inflammatory cells. In addition, in resistant strains, LPS often enhanced and never depressed the inflammatory response to other phlogistic agents. These studies demonstrate that the genetic regulation of the inflammatory responses to LPS also controls the anti-inflammatory effects of LPS. These responses may be another relevant parameter in determining strain susceptibility to LPS lethality.
Analysis of Ir gene function using monoclonal antibodies: independent regulation of GAT and GLPhe T cell responses by I-A and I-E subregion products on a single accessory cell population.
Authors: Nepom J T , Benacerraf B , Germain R N
Published: December 2022

T cell proliferative responses to the synthetic polypeptides GAT and GLPhe are under Ir gene control. GAT responses are regulated by gene(s) in the I-A subregion, and GLPhe responses are controlled by a pair of complementing genes mapping to the I-A and I-E subregions. We demonstrate that monoclonal antibody to the I-A gene product inhibits GAT proliferation but not the GLPhe response, whereas a monoclonal antibody to the I-E associated Ia-7 determinant inhibits GLPhe but not GAT proliferation, which indicates independent involvement of each Ia determinant in antigen presentation for the T cell response to these antigens. Use of the same subregion-specific monoclonal antibodies in complement-dependent lysis demonstrates that the antigen-presenting cells for GAT and GLPhe express both I-A and I-E products. The possibility that an Ia subregion-specific "self-receptor" functions on the reactive T cells as a regulatory element is discussed.
Ultraviolet light induces tumors with both unique and host-associated antigenic specificities.
Authors: DeWitt C W
Published: December 2022

Murine tumors induced by ultraviolet light (UV) are immunogenic in syngeneic and semi-syngeneic hosts, evoking antibody of several different specificities. Cytotoxic antibody specific for the immunizing syngeneic tumor (tumor-specific antigen) comprises the early response and a minor portion of the later response of C3H and C3H.SW mice. It is the primary specificity to which C57BL/6 and C57BL/10 hosts respond. The major portion of the antibody produced by C3H and C3H.SW against syngeneic tumors cross-reacts strongly with other tumors, both UV and chemically induced, arising in C3H and C3H.SW but not in B6.H2k, C57BL/6, C57BL/10, or (C3H X B6)F1. Normal adult cells or embryonic fibroblasts do not cross-react with the antisera. These results are interpreted as evidence for the involvement of a host component (non-MHC) in this tumor-associated antigen (TAA). (C3H X B6)F1 and (C3H X BALB/c)F1 hosts respond to C3H tumors with antibody with cross-reactive specificities identical to those of the C3H and C3H.SW hosts. thus detecting the TAA(C3H) specificity. (C3H X B6)F1 hosts respond to syngeneic F1 tumors, however, with a totally cross-reactive antibody that is interpreted as evidence for the existence of a common antigen in addition to the evident immune response control. An undetected TAA (B6) specificity in the (C3H X B6)F1 tumors is speculatively proposed.