bluuebunny/crossref_metadata_2025 huggingface dataset:
Production of Migration Inhibition Factor by Spleen Cells of Normal Rats upon Culture in vitro with Tumor Cells and Cells Expressing Endogenous Virus
Authors: Sharma J. M. , Herberman R. B. , Djeu Julie Y. , Nunn Myrthel E.
Published: December 2022

Spleen cells from normal rats of W/Fu, Fischer, Sprague-Dawley, and BDIX strains produced migration inhibition factor (MIF) upon culture in vitro with puromycin-treated rat cell lines expressing leukemia viruses or endogenous C-type viruses. Brown Norway rats were almost invariably unreactive. The MIF reactivity of most normal rats did not appear to be due to environmental stimuli because germfree rats were also positive. There was no apparent age restriction for expression of MIF activity; spleen cells from 1-week-old rats produced detectable levels of MIF, and this reactivity was also found in the oldest tested rats, at 34 weeks of age. In addition to spleen cells, lymphoid cells from the blood, peritoneal cavity, and lymph nodes, but not thymus, also were able to produce MIF in response to tumor cells. When (C58NT)D tumor cells, which were routinely used as the stimulators of MIF production, were inoculated into W/Fu rats, the characteristics or extent of preexisting reactivity were not altered. However, spleen cells of most rats bearing progressively growing (C58NT)D tumors did not show a MIF response in vitro.

MIF production by spleen cells was dependent upon the presence of macrophages. Spleen cells depleted of phagocytic and adherent cells were unable to produce MIF upon incubation by tumor cells; however, the MIF response could be restored by adding allogeneic or syngeneic macrophages to the depleted spleen cells. The helper role of macrophages in the generation of MIF by spleen cells did not appear to involve antigen presentation because direct contact between macrophages and the responder cells was not necessary, and soluble factor(s) released by macrophages could substitute for the macrophage helper function.

The effector cells of MIF production were nonadherent, lacked detectable Fc receptors for IgG, and were resistant to lysis by anti-T cell serum plus complement. Because the MIF response was present in apparently normal rats, this appears to be an expression of natural immunity that may be analogous to natural killer cell activity.
Analysis of B Cell Activation Requirements with TNP-Conjugated Polyacrylamide Beads
Authors: Mond J. J. , Stein K. E. , Subbarao B. , Paul W. E.
Published: December 2022

TNP-polyacrylamide beads of varying epitope density were used as antigens to stimulate normal adult spleen cells, neonatal spleen cells, and spleen cells from mice with the CBA/N immune defect. Lymphocytes from immune defective (CBA/N × DBA/2)F1 male mice or normal 2- to 3-week-old (C57BL/6 × DBA/2) F1 mice were unable to respond to TNP-polyacrylamide beads of low epitope density but responded quite well to high density TNP-polyacrylamide beads. Furthermore, immune defective F1 male spleen cells were shown to be unresponsive to TNP-Dextran yet mounted significant responses to TNP-Sephadex, an insoluble analog of TNP-Dextran.

Use of an antiserum with anti-Lyb7.1 activity, which recognizes a mature subset of B cells, also pointed to qualitative differences in the stimulatory activities of highly substituted TNP-polyacrylamide beads and lightly substituted TNP-beads. This antiserum was able to inhibit responses to the low epitope TNP-polyacrylamide beads, which presumably activate a mature subset of B cells, but did not block responses to the highly conjugated TNP-beads, which appear to activate both immature B cells and mature B cells.

In the course of these studies it became clear that TNP-polyacrylamide bead is not a truly thymus-independent (TI) antigen. Although it is able to stimulate significant responses in nude spleen cells, it is an ineffective immunogen when cultured with cells extensively depleted of T cells.
The Suppression of Mitogen- and Alloantigen-Stimulated Peripheral Blood Lymphocytes by Cultured Human T Lymphocytes
Authors: Maca Richard D. , Bonnard Guy D. , Herberman Ronald B.
Published: December 2022

Cultured T lymphocytes (CTC), which were grown in medium containing a growth-promoting factor from supernatants of pooled, allogeneic, PHA-stimulated PBL, were examined for their ability to suppress mitogen-induced proliferative responses or MLC reactions. This suppressor activity was assayed by adding the CTC to either Con A- or PHA-treated lymphocytes or to a mixed lymphocyte culture. The incorporation of 3HTdR into nuclear protein was then quantitated at the end of the culture period and compared to the control 3HTdR uptake values. The addition of CTC significantly suppressed thymidine incorporation in both the mitogen- and alloantigen-stimulated cultures. This suppressed thymidine uptake was not due to competitive inhibition by cold thymidine released into the culture medium by the CTC. Likewise, this suppressor activity could not be accounted for by a “crowding” effect of the added CTC or by the depletion of nutrients from the culture medium by metabolizing CTC. CTC autologous to the responding normal lymphocytes were as effective in suppressing the mitogen and alloantigen responses as allogeneic CTC, indicating that the observed suppression by CTC could not be explained by the cytotoxicity by alloantigen-sensitized or polyclonal-activated CTC. Viable and irradiated CTC were equally effective in suppressing the Con A response. However, in the response to allogeneic cells, irradiating the CTC completely abrogated the suppressor effect. This model system provides a good approach for studying suppressor activity of human T lymphocytes that are not contaminated by other types of cells with suppressor potential, such as monocytes. In addition, this system should provide a means to evaluate ways by which this T cell suppressor activity can be modulated.
Gene Complementation in the T Lymphocyte Proliferative Response to Poly (Glu57Lys38Tyr5): Evidence for Effects of Polymer Handling and Gene Dosage
Authors: Schwartz Ronald H. , Merryman Carmen F. , Maurer Paul H.
Published: December 2022

This paper explores the difficulties encountered in demonstrating Ir gene complementation in the immune response to the synthetic terpolymer, poly(Glu57-Lys38Tyr5) [GLT5]. First, the method by which the polymer was put into solution influenced the outcome because some determinants on GLT5 were found to be sensitive to exposure to alkaline conditions. Second, the dose of antigen used for immunization was shown to be critical since mice of the H-2q haplotype failed to respond to low doses of GLT5. Third, prolonged storage of dialyzed and lyophilized GLT5 led to alterations in the solubility properties of the polymer with consequent loss of some of the antigenic determinants. Finally, differences in the genetic make up of the responding strains affected the overall response. Each responder haplotype recognized different determinants on the polymer, or the same determinant in different ways, such that variations in polymer handling appeared to influence selectively the immunogenicity and antigenicity of GLT5 for each strain. Furthermore, the critical (B10 × B10.A)F1 strain, whose response was required to demonstrate conclusively complementation, failed to respond well to certain altered preparations of the polymer in which the determinants being recognized were present in low concentration. This failure, in contrast to the high responsiveness of the genetically similar B10.A(5R) strain, was shown by the low response of [B10.A(4R) × B10.A(5R)] F1 mice to result from a gene dosage effect. However, under optimal conditions of antigen handling, (B10 × B10.A)F1 mice did respond well to GLT5, demonstrating that two complementing MHC-linked Ir genes are involved in the recognition of certain determinants on this polymer.
Secretory Responses of Human Neutrophils: Exocytosis of Specific (Secondary) Granules by Human Neutrophils during Adherence in vitro and during Exudation in vivo
Authors: Wright Daniel G. , Gallin John I.
Published: December 2022

Human neutrophils have been shown to release lysozyme spontaneously when incubated in vitro in the presence of physiologic concentrations of calcium. This spontaneous release of lysozyme, stored in both the azurophil and specific granules of neutrophils, reflects a selective mobilization and exocytosis of the specific granules, for it occurs without release of granule products located strictly in the azurophil granules (i.e., β-glucuronidase). Calcium-dependent lysozyme release without β-glucuronidase release by human neutrophils is significantly enhanced when the cells are exposed to surfaces to which they avidly adhere (nylon fibers and glass). Release of lysozyme, without a comparable release of β-glucuronidase, also occurs when human neutrophils migrate across cellulose nitrate micropore filters in vitro in response to a chemotactic stimulus. These phenomena in vitro appear to mimic events that occur in vivo when human neutrophils leave the circulation to become exudate cells. Specific granule products (lysozyme and B12-binding protein) but not azurophil granule products appear in the extracellular fluid of experimental inflammatory lesions in vivo coincident with the influx of exudate neutrophils. Also, analysis of the granule content of exudate neutrophils, by granule separation techniques and by cytochemistry and electronmicroscopy, demonstrates a loss of specific granules but not azurophil granules from exudate cells when compared with peripheral blood neutrophils studied concurrently. Results of these studies support the concept that neutrophil-specific granules function in part like the storage granules of secretory cells and that mobilization and exocytosis of these granules by neutrophils are intrinsic to normal inflammatory responses. The release of specific granule products may contribute to extracellular killing or inhibition of microorganisms, but may also provide mechanisms by which neutrophils influence other inflammatory blood and tissue cells.
The LT System in Experimental Animals
Authors: Hiserodt John C. , Tiangco Gerry J. , Granger Gale A.
Published: December 2022

The present studies investigate the physicochemical and immunologic properties of cell-lytic molecules released in vitro by nonadherent C57/BL/6 splenocytes or nylon wool-enriched T cell populations activated on monolayers of PHA coated L-929 cells. The findings reveal cell-lytic molecules released by these lymphoid cells are physically heterogeneous. These molecules can be separated by gel filtration into similar m.w. classes previously observed for human LT molecules. Three major classes, termed complex (Cx) (>200,000 d), α heavy (αH) (110 to 140,000 d)α light (αL) (60 to 90,000 d), and two minor classes, β, at 40 to 50,000 d and γ at 10 to 20,000 d were observed. Chromatography of supernatants in high ionic strength buffers dissociates Cx and αH to the smaller m.w. αL form. This evidence suggests that Cx and αH MW classes are physically related to the smaller m.w. αL class. Fractionation of the αH m.w. LT class by DEAE or PAGE resolved these molecules into additional distinct subclasses. Antisera were made against fresh serum-free whole supernatants (anti-WS) or rechromatographed Ultrogel fractions containing αH molecules (anti-αH). Anti-αH and anti-WS react with all m.w. classes of murine LT molecules, indicating these various forms are immunologically related. These antisera do not react with LT molecules obtained from several other animal species or with “nonspecific” intracellular toxins, e.g., lysozomal enzymes, present in normal PMN or phagocytic cells.

These data indicate that materials with cell-lytic activity present in these culture supernatants are LT molecules, because: a) certain m.w. forms observed are similar to those reported previously, and b) these various m.w. forms are all physically and immunologically interrelated. These studies also indicate that murine LT molecules like human LT molecules are heterogeneous, but appear to comprise a system of subunits, in which the large m.w. form may dissociate into the smaller m.w. forms.
Inhibition of in vitro Cytotoxic Responses by BCG-Induced Macrophage-Like Suppressor Cells
Authors: Klimpel Gary R. , Okada Masaji , Henney Christopher S.
Published: December 2022

The ability of adherent, macrophage-like spleen cells from BCG-infected C57BL/6 mice to suppress primary cytotoxic responses to alloantigen in vitro has been studied. Suppression was dependent on a viable suppressor cell, for both heating (56°C 30 min) and glutaraldehyde treatment (0.1%, 2 min) ablated the suppressive action. Membrane preparations were not suppressive. Suppression was reversible and independent of the suppressor cell's Fc receptor sites. Thus, suppressor cell populations pretreated with heat-aggregated IgG or bound to immobilized immune complexes were still capable of suppressing the cytotoxic response. We could find no evidence that the suppressor cells function through the production of a soluble mediator.

The principal mode of action of the suppressor cell seemed to reflect a capacity to modify the production and action of a mediator whose production was dependent upon helper (Lyt 1+) T cells. This alloantigen-induced mediator, which augments the proliferation and/or differentiation of cytotoxic T cells, was not produced in mixed lymphocyte cultures containing spleen cells of BCG-infected animals nor by normal spleen cells in the presence of the BCG-induced suppressor cells. Under appropriate conditions, mediator produced from mixed lymphocyte cultures was able to reverse the suppression of primary cytotoxic responses by BCG-induced suppressor cells.
Re-Expression of Nonglycosylated Surface IgA in Trypsin-Treated MOPC 315 Plasmacytoma Cells
Authors: Hickman Scot , Wong-Yip Y. P.
Published: December 2022

The importance of glycosylation for the re-expression of surface immunoglobulin in trypsin-treated MOPC 315 plasmacytoma cells was examined by using tunicamycin, an antibiotic that prevents glycosylation by inhibiting the formation of N-acetylglucosamine-lipid intermediates. Tunicamycin greatly inhibited the secretion of nonglycosylated MOPC 315 IgA in trypsin-treated cells. Two hours after trypsin treatment, there was an 80% inhibition of secretion as measured by immunoprecipitation assays of biosynthetically labeled immunoglobulin. However, tunicamycin had no effect on the time course of re-expression of surface IgA in these cells as measured by TNP-sheep erythrocyte rosette formation and [125I] TNP-albumin binding to the plasmacytoma cells. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of 125I-labeled cell surface IgA re-expressed in the presence of tunicamycin revealed a protein with an apparent m.w. identical to nonglycosylated MOPC 315 α-chains, further suggesting that nonglycosylated surface IgA was being inserted into the plasma membrane. This protein did not bind to concanavalin A-Sepharose. These data suggest that in MOPC 315 plasmacytoma cells, glycosylation is necessary for immunoglobulin secretion but not for immunoglobulin expression at the cell surface.
Cross-Priming of Murine B Cells with TNP Conjugates of Hemocyanin and Ficoll: Characteristics of Primed B Cells Responding to Both Antigens
Authors: Umetsu D. T. , Chapman-Alexander J. M. , Thorbecke G. J.
Published: December 2022

There is strong evidence that separate subsets of B cells respond to primary challenge with TNP-Ficoll (TI.2) and with TNP-KLH (TD). Although TD antigens can prime for responses to TI antigens, it has been suggested that, at least in vitro, distinct subsets respond to TI.2 and TD antigens even after such priming. The present experiments on adoptive secondary immune responses in LAF1 mice suggest that a single population of primed cells may respond to both antigens. This conclusion is based on the findings that 1) High-affinity 7S antibody-producing cells primed by TNP-KLH can be challenged by TNP-Ficoll even after removal of T cells; 2) Priming with TNP-Ficoll induced 2- to 4-fold increased responsiveness to itself and to TNP-KLH, although the maturation with respect to affinity distribution of these IgG responses is significantly less than after priming with TNP-KLH; 3) TNP-KLH primed-cells never give higher responses upon challenge with both antigens simultaneously than to one of these antigens injected alone; 4) Primary and secondary responses to both antigens are reduced to a similar degree after pretreatment of cells with anti-Ia and C; 5) Receptor blockade of TNP-KLH primed cells by incubation with TNP-SIII reduces responses to TNP-KLH by 77 to 90% and to TNP-Ficoll by 60 to 70%. The findings are interpreted as supporting the view that B cell subpopulations responding to TI.2 and TD antigens represent stages of differentiation of a single B cell lineage that are overlapping rather than distinct subsets in vivo.
Activation of Suppressor T Cells in Human Autologous Mixed Lymphocyte Culture
Authors: Smith J. Bruce , Knowlton Regina P.
Published: December 2022

Co-culture of autologous T and mitomycin-C treated B cells results in increased DNA synthesis in the responding T cells. T cells thus activated in AMLC exerted suppressive effects on both the proliferative and cytotoxic responses of fresh unstimulated T cells to allogeneic cells in MLC. The suppressor cells generated are sensitive to treatment with mitomycin-C. AMLC-activated cells treated with mitomycin-C failed to suppress both cytotoxicity and proliferation in fresh primary MLC. It appears that the AMLC reaction reflects an immunologic homeostatic mechanism. Since this reaction is defective in patients with CLL and SLE and the homologous mouse syngeneic MLC is defective in NZB mice, the failure of this T-B interaction may be related to the pathogenesis of certain lymphoproliferative and autoimmune disorders.
X-Linked B Lymphocyte Defect in CBA/N Mice
Authors: Scher Irwin , Berning Alice K. , Asofsky Richard
Published: December 2022

The primary thymus-dependent IgG anti-sheep red blood cell (SRBC) response of (CBA/N × DBA/2)F1 male mice was <10% of that of immunologically normal (CBA/N × DBA/2)F1 female mice. Adoptive transfers of activated T cells (ATC) and either bone marrow or anti-ϑ-treated spleen cells from F1 mice into lethally irradiated recipients demonstrated that the functional helper-cell activity of ATC from immune-defective and normal F1 mice was equivalent. By contrast, B lymphocytes of (CBA/N × DBA/2)F1 male mice gave very poor or absent IgG anti-SRBC responses in this system, whereas the response of B cells from immunologically normal F1 mice was excellent. The poor response of (CBA/N × DBA/2)F1 male B cells was not improved by providing additional T cell help, suggesting that these cells were refractory to a large proportion of this help. Furthermore, increasing the number of donor B cells from immune-defective mice failed to influence materially the IgG anti-SRBC response. Although the response of (CBA/N × DBA/2)F1 male primed B cells was considerably improved over that of unprimed cells, it remained less than the primary response of immunologically normal F1 mice. The presence of a low but detectable SRBC response in (CBA/N × DBA/2)F1 male mice suggests that the B cells of these mice, and by inference those of normal mice, can be triggered by antigen in the presence of ATC through a mechanism that is independent of Lyb 3+, 5+ B cells. The large increase in the response of (CBA/N × DBA/2)F1 male B cells to SRBC after priming suggests that Lyb 3+, 5+ B cells are not necessary for the generation of memory B cells.

An interesting difference in the environments of (CBA/N × DBA/2)F1 vs (DBA/2 × CBA/N)F1 male mice was demonstrated by reconstituting these mice with mixtures of ATC and B cells after lethal irradiation. In all cases, the IgG responses of the immune-defective F1 mice were significantly less than those of the immunologically normal mice. Mechanisms that could explain these findings are discussed.
Polymorphonuclear Leukocyte-Mediated, Antibody-Dependent, Cellular Cytotoxicity against Tumor Cells: Dependence on Oxygen and the Respiratory Burst
Authors: Hafeman Dean G. , Lucas Zoltan J.
Published: December 2022

Experiments were done to determine 1) whether the respiratory burst of superoxide anion (O2-) production in polymorphonuclear leukocytes (PMN) is triggered during antibody-dependent killing of tumor cells and 2) whether O2- production is essential for cytotoxicity. Three parameters of the respiratory burst (1-14C-glucose oxidation, oxygen consumption, and O2- release) were increased 2.5- to 7.3-fold during killing of antibody-primed tumor cells by human PMN. Added catalase and superoxide dismutase did not inhibit lysis, possibly because these enzymes were unable to diffuse into the inter-plasma-membrane space between killer and target cells. Evidence for an O2- requirement for cytotoxicity was the fact that concentrations of amobarbital or phenylbutazone sufficient to inhibit the cyanide-insensitive respiration of PMN also inhibited cytotoxicity. Also, hypoxic conditions inhibited cytotoxicity from 29 to 73%. The requirement for oxygen was most likely related to O2- generation and not mitochondrial respiration since cyanide and azide, which inhibit mitochon drial respiration, increased cytotoxicity.
Phenotypic and Functional Distinctions between the TH2+ and JRA+ T Cell Subsets in Man
Authors: Reinherz Ellis L. , Strelkauskas Anthony J. , O'Brien Carmeline , Schlossman Stuart F.
Published: December 2022

Prior work has demonstrated the existence of distinct human peripheral blood T cell subsets by utilizing heterologous as well as autoimmune antisera. In the present study, the relationship between the TH2+ and JRA+ T cell subsets was examined. T cells were purified with Sephadex G-200 anti-F(ab)2′ affinity chromatography and E-rosetting technique, and subsequently fractionated into TH2+ and TH2- subsets by utilizing indirect immunofluorescence on FACS. Approximately 40 to 45% of the TH2- subset was shown to be JRA+, whereas less than 5% of the TH2+ subset was JRA+. In reciprocal studies, T cells were fractionated into JRA+ and JRA- subsets and reacted with heterologous antisera with anti-TH2+ specificity and indirect immunofluorescence. FACS analysis demonstrated that the JRA+ population contained no TH2+ T cells. In contrast, the JRA- population contained TH2+ T cells and accounted for the entire TH2+ subset found in the unfractionated T cell population. Functional studies showed that the TH2+ subset, and not the JRA+ subset, contain the effector population for cell-mediated lympholysis. It is concluded that the TH2+ and JRA+ T cell subsets define distinct and different T cell populations in man.
Inhibition of Methylation by Adenosine in Adenosine Deaminase-Inhibited, Phytohemagglutinin-Stimulated Human Lymphocytes
Authors: Johnston Jeffrey M. , Kredich Nicholas M.
Published: December 2022

Adenosine toxicity was studied in phytohemagglutinin-stimulated, normal human, peripheral lymphocytes in order to determine whether inhibition of S-adenosylmethionine (AdoMet)-mediated transmethylation reactions might contribute to the immune defect associated with heritable deficiency of the enzyme adenosine deaminase (ADA). Cells were treated with 20 µM erythro-9-(2-hydroxy-3-nonyl)adenine to inhibit ADA and with 20 µM uridine to prevent the pyrimidine starvation known to be caused by adenosine in this system. The further addition of 25 to 100 µM adenosine inhibited the uptake of radiolabeled thymidine and leucine into acid precipitable material, and also increased intracellular levels of S-adenosylhomocysteine (AdoHcy) from 3- to 8-fold. The formation in these experiments of AdoHcy, a potent inhibitor of AdoMet-mediated transmethylation reactions, presumably occurs through the action of S-adenosylhomocysteine hydrolase. Although the physiologic function of this enzyme is to hydrolyze AdoHcy, the reversibility of the reaction and an equilibrium constant favoring synthesis allow net synthesis of AdoHcy under conditions of adenosine excess. Inclusion of 100 to 200 µM L-homocysteine thiolactone in our experiments enhanced AdoHcy accumulation and also potentiated the inhibitory effects of adenosine on radiolabeled thymidine and leucine uptake. Cytotoxic concentrations of adenosine also caused significant inhibition of in vivo DNA methylation, a process known to be AdoMet-mediated, and a good correlation between this effect and AdoHcy levels was noted. These findings indicate that adenosine toxicity in cultured human lymphocytes is at least in part due to AdoHcy accumulation and resultant inhibition of essential AdoMet-mediated methylation reactions. The relevance of these findings to the pathogenesis of the immune defect in ADA-deficient patients is not established but deserves consideration.
The Regulation of Thymidine Secretion by Macrophages
Authors: Stadecker Miguel J. , Unanue Emil R.
Published: December 2022

The secretion of thymidine by mononuclear phagocytes was correlated with the activity of the enzyme thymidine kinase (TK). Macrophages cultured in regular tissue culture medium released thymidine and did not express TK. However, when macrophages were incubated with medium conditioned by L cells, they expressed TK, incorporated 3H thymidine into trichloroacetic acid precipitable material, and ceased to secrete the nucleoside. Furthermore, replicating P388/D1 cells were induced to secrete thymidine by inhibiting TK with d-glucosamine. These results have demonstrated an inverse relationship between thymidine secretion and the expression of TK. They suggest that thymidine secretion by macrophages may be attributed to their lack of TK activity.
Mononuclear Cells Contain Human Transfer Factor as Assayed Locally on the Skin of Dogs
Authors: Tomar Russell H. , John Patricia A.
Published: December 2022

We prepared transfer factor from mononuclear (M) cells and polymorphonuclear (PMN) cells by sieving cell lysates through a P-2 Biogel column. By assaying column fractions by their ability to induce a local delayed-type reaction to antigen in canine skin, we determined that mononuclear cells most consistently contained skin activity. The elution patterns from P-2 Biogel of PMN or M-enriched preparations were less complex and much different from that seen with whole leukocytes.
Circadian Rhythm in Activity of Lympholytic Natural Killer Cells from Spleens of Fischer Rats
Authors: Fernandes G. , Carandente F. , Halberg E. , Halberg F. , Good R. A.
Published: December 2022

Three and one-half-week-old male inbred Fischer rats housed individually at 24 ± 1°C with Purina Chow and water ad libitum were randomly assigned to six different rhythmometry chambers, each lighted daily for 12 hr and darkened the following 12 hr. The timing of these regimens differed from one chamber to the next by 4 hr (i.e., one chamber was illuminated from 0600 to 1800, the next from 1000 to 2200, etc.). Thus, completely light-dark synchronized animals sampled from the six chambers at any given clock time were intended to represent, in relation to the onset of the daily light span, six different circadian stages, differing from each other by 4 hr. After 2-week standardization in the chambers, rectal temperature was measured and the animal killed, its spleen collected aseptically in RPMI 1640 medium containing 10% fetal calf serum, and the activity of natural killer (N.K.) cells determined in a direct 51Cr release assay. Spleen cells of 6-week-old rats were effector cells and RL♀1 mouse lymphoma cells in tissue culture were target cells. Two experiments carried out, each on 3 successive days in November 1977 and February 1978, using subtotals of 24 rats at each lighting stage; i.e., a total of 144 rats for six intended circadian stages, revealed by cosinor analysis a circadian rhythm in N.K. cell activity and rectal temperature. The circadian recetal temperature acrophase may be a marker with similar timing of high murine N.K. cell activity. In the context of a budding chronoimmunology, these findings extend the scope of a spectrum of bioperiodic neuroendocrine influences on lymphoid activity and body resistance.
The Role of Cyclic AMP in Modulating Cytotoxic T Lymphocytes
Authors: Plaut Marshall
Published: December 2022

Cytotoxic T lymphocytes (CTL) were generated in the spleen by immunizing C57BL/6 mice (H-2b) i.p. with allogeneic P815 cells (H-2d), or by culturing C57BL/6 spleen cells in vitro with mitomycin C-treated P815 cells. Agents that increase cyclic AMP levels, including histamine, prostaglandin E2, dibutyryl cyclic AMP, and theophylline, markedly inhibited the activity of in vivo-generated CTL. In contrast, these agents had little inhibitory effect on the activity of in vitro-generated CTL. The inhibition by cyclic AMP-active agents was not altered by varying lymphocyte:target cell ratios or assay times, or by removing dead cells. The differences in inhibition by histamine and dibutyryl cAMP persisted even in the presence of high concentrations of the cAMP phosphodiesterase inhibitors, theophylline and Ro 20-1724. Inhibition by mixtures of in vivo- and in vitro-generated CTL populations was the weighted average of the inhibition of each separate population. When in vivo-generated CTL were restimulated in vitro with antigen, within 24 hr, the inhibition by cAMP-active agents of the resultant CTL was no greater than inhibition of in vitro-generated, primary CTL. Thus, culture induces nonspecific desensitization of CTL to cyclic AMP-active agents, possibly by altering biochemical events in CTL such that their cytotoxic mechanism is disassociated from the cAMP-mediated regulatory step(s) that are present in in vivo populations.
Cloning of B Cells Positive or Negative for Surface IgD
Authors: Layton Judith E. , Pike Beverley L. , Battye Frank L. , Nossal G. J. V.
Published: December 2022

A direct experimental approach has been used to test two hypotheses for the function of B lymphocyte surface IgD (sIgD). The first hypothesis is that the presence of sIgD distinguishes cells that respond to T-dependent antigens from those that respond to T-independent antigens and the second is that the presence of sIgD is responsible for resistance to tolerance induction. The approach used was to separate sIgD+ cells from sIgD- cells in the fluorescence-activated cell sorter and to use cloning assays in which the precursor activity is linearly related to the number of cells assayed, to test the activity of separated populations. Mice at 2½ weeks of age were used in most experiments to provide spleen cells in which there was an equal proportion of sIgD+ and sIgD- B cells. In this paper, the immune response and tolerance induction were examined in T-independent assay systems. Both sIgD+ and sIgD- cells responded to a T-independent antigen (FLU-POL) and mitogens, with the better response obtained from the sIgD+ population. The same degree of tolerance susceptibility was found in sIgD+ and sIgD- populations from either 2½-week-old or adult mice. The conclusion from these observations is that the T-independent response is not restricted to sIgD- B cells and also that resistance to tolerogenesis in a T-independent assay system is not caused by acquisition of IgD.
Fc Receptors for IgA on Human B, and Human Non-B, Non-T Lymphocytes
Authors: Lum Lawrence G. , Muchmore Andrew V. , O'Connor N. , Strober W. , Blaese R. Michael
Published: December 2022

Recently, receptors for IgA were demonstrated on subpopulations of human T lymphocytes. In this report, TNP-modified ox erythrocytes coated with the IgA myeloma MOPC-315 were used to detect IgA receptor-bearing lymphocytes within the human non T cell lymphocyte population. A mean of 5.3% (range 2.9 to 12.4%) of Erosette negative human lymphocytes bound IgA-coated indicator cells. Blocking studies with soluble IgA, IgG, and IgM demonstrated that the IgA receptors on the non-T cell populations were separate and distinct from the Fc-receptors for IgG and IgM. Fractionation of the non-T lymphocytes on anti-human (Fab)2 columns into sIg+ and sIg- populations or by rosetting with EAC to provide CRL+ and CRL- populations demonstrated that Fc-IgA receptors were present on a subpopulation of sIg+, CRL+ lymphocytes, and also on sIg- (non-T, non-B) lymphocytes.
Interaction between T Cells and Non-T Cells in Suppression of Cytotoxic Lymphocyte Responses
Authors: Truitt Gary A. , Dennison David K. , Rich Robert R. , Rich Susan Solliday
Published: December 2022

Generation of cytotoxic T lymphocytes (CTL) in mixed leukocyte cultures was suppressed by a factor elaborated by alloantigen-activated T cells. This suppressor factor, CTL-TsF, in contrast to a factor that suppresses proliferative responses in mixed leukocyte reactions (MLR-TsF), was effective only when added during the first 24 hr of a 6-day-culture period. Moreover, removal of CTL-TsF 24 hr after culture initiation failed to restore CTL responses. CTL activity could be rescued from suppressed cultures, however, by addition of 2-mercaptoethanol on days 3 or 4. Similarly, transfer of nonadherent cells at 3 or 4 days from cultures treated with CTL-TsF to cultures of adherent cells initiated in control factor restored CTL responses. Mixing experiments with cells pulsed with CTL-TsF for 4 hr at culture initiation identified a target of CTL-TsF as a Thy-1 negative cell that was adherent to plastic and to Sephadex G-10. Suppression was not due to interference with physiologic accessory cell function, but more likely was accomplished via a negative signal from CTL-TsF-pulsed cells. The results thus suggest that CTL-TsF acts early, but reversibly, in the CTL differentiative process via a second suppressor effector cell, possibly a macrophage.
Effects of Concanavalin A-Induced Cells on the Proliferative Response of T Cells
Authors: Mayumi Mitsufumi , Yoshida Takami , Shinomiya Keisuke , Nishikawa Shin-Ichi , Hirata Takeo , Izumi Takateru , Mikawa Haruki
Published: December 2022

Effects of Con A-induced human mononuclear cells on the proliferative response of peripheral T cells were examined by using TNP-modified autologous lymphocytes as stimulator cells. Cells induced by incubation with Con A contained both suppressor cells and amplifier cells. The former were induced from nylon woolnonadherent T cells and these precursor cells were sensitive to mitomycin treatment. On the other hand, amplifier precursor cells were nylon wool-nonadherent T cells and were resistant to mitomycin treatment. Cell proliferation was required for the induction of suppressor cells but not for the induction of amplifier cells. Con A-induced suppressor effector cells were both nylon wool-adherent and nonadherent cells, on the contrary, Con A-induced amplifier effector cells were nonadherent cells. A small number of macrophages enhanced the suppressive activity of nonadherent T cells when added at the induction phase of suppressor T cells.
The Association of Immune Responsiveness, Mixed Lymphocyte Responses, and Ia Antigens in Natural Populations of Norway Rats
Authors: Shonnard John W. , Davis Bridgett K. , Cramer Donald V. , Radka Susan F. , Gill Thomas J.
Published: December 2022

The major histocompatibility complex (MHC) of the rat has been separated into two regions. The A region contains genes that code for the classical histocompatibility (Ag-B/H-1) antigens, and the B region codes for the MLR, Ia antigens, and immune response to synthetic polypeptides. We have examined the relationship between MLR, Ia antigens, and the immune response to poly(Glu52Lys33Tyr15) in inbred rats and in a population of wild rats captured in a single location within the city of Pittsburgh. Antisera that define Ia antigenic specificities association with the RT1.Ba, RT1.Bl, and RT1.Bn MLR phenotypes in inbred rats detected the same MLR phenotype in each of 48 individual wild rats. This close association between Ia specificities and MLR phenotypes in a randomly breeding wild population suggests that the genes controlling the expression of these phenotypic strains are either tightly linked or that Ia antigens represent a stimulating determinant for the MLR.

In the same population of wild rats, distinct high and low immune responses to the synthetic polypeptide poly(Glu52Lys33Tyr15) could be detected. Breeding studies demonstrated that the immune response to poly-(Glu52Lys33Tyr15) segregated with the MHC. There was a close association between high immune responsiveness and the RT1.Ba (Ia.4) MLR phenotype, and low responsiveness was usually associated with either the RT1.B1 (Ia.1) or RT1.Bn (Ia.3) MLR phenotypes. These same MLR and immune response associations are observed in inbred animals.

The close association between alleles at different genetic loci in both the A and B regions of the rat MHC in natural populations is very similar to that reported for specific HLA haplotypes in man, suggesting that the rat may prove to be an important experimental model for studies involving the population dynamics of the genes in the MHC.
Fractionation of Human Lymphocytes with Plant Lectins
Authors: Boldt David H. , Lyons Ruth D.
Published: December 2022

We have fractionated human peripheral blood lymphocytes (PBL) into distinct subclasses based upon differential adherence to the plant lectins wheat germ agglutinin (WGA) and Lens culinaris lectin (lentil-PHA), derivatized to gelatin surfaces in plastic tubes and Petri dishes. Fifteen to forty percent of PBL were adherent to WGA-derivatized gelatin. Binding studies with 125I-WGA showed that, compared to unfractionated PBL which bound 31.9 × 106 WGA molecules/cell, adherent cells bound more (44.1 × 106 molecules/cell), and nonadherent cells bound less (20.4 × 106 molecules/cell) WGA. By contrast, there were no differences among these groups for binding of other plant lectins. When lentil-PHA-derivatized gelatin was used, 25 to 50% of PBL were adherent, and, as previously reported, adherent cells bound approximately 4 times as many lentil-PHA molecules as nonadherent cells, whereas there were no differences in binding of other lectins. Sequential fractionation studies indicated that the WGA- and lentil-PHA-selected subclasses were separate groups. The percentage of T and B lymphocytes and macrophages did not differ among the subclasses. WGA-selected and lentil-PHA-selected lymphocyte subpopulations responded differently to stimulation in vitro by a panel of plant mitogens. Human PBL contain a spectrum of cell subclasses with different lectin-binding capacities. These subclasses are functionally unique.
Immune Response in Subacute Sclerosing Panencephalitis: Reduced Antibody Response to the Matrix Protein of Measles Virus
Authors: Wechsler Steven L. , Weiner Howard L. , Fields Bernard N.
Published: December 2022

Immune precipitation was used to study the humoral immune response of patients with subacute sclerosing panencephalitis (SSPE). Patients with SSPE have a progressive infection of the CNS by measles or a measles variant despite high serum antibody levels to measles virus as measured by standard serologic techniques. However, when the antibody response to individual measles virus proteins was measured, we found a striking reduction in the ability of sera from patients with SSPE to precipitate the matrix (M) protein as compared to the precipitation of the M protein by sera from normal adults who had natural measles infection in childhood, or by convalescent sera obtained 3 to 5 weeks after a naturally occurring measles infection. The decreased antibody response to the M protein in sera from patients with SSPE occurred despite a vigorous antibody response to the other viral proteins, suggesting a selective defect in the production of antibody to a single viral protein. The reduced anti-M antibody in sera from patients with SSPE was demonstrated whether immune precipitation was performed with wild-type measles virus or SSPE virus proteins. These results suggest that in SSPE only small amounts of the M protein are produced. This results may help explain how measles virus persists in the central nervous system of patients with SSPE.
Manifestations of Cellular Immunity in the Rat After Prolonged Asbestos Inhalation
Authors: Miller Klara , Weintraub Zeona , Kagan Elliot
Published: December 2022

A number of recent studies have suggested that the aleolar macrophage plays a crucial role in the development of asbestosis. In the current study the possible alterations of the cell-mediated immunologic mechanisms by asbestos-exposed macrophages were examined. Two inbred rat strains (BD-IX and AGUS) were exposed to UICC crocidolite asbestos by inhalation for extended periods. Alveolar macrophages and nylon column-purified T cell-enriched splenic lymphocytes were co-cultured (in both syngeneic and allogeneic situations) for 1 or 22 hr in vitro. Parallel studies were also performed on cell populations from control (nondusted) rats. After 1-hr culture significantly greater numbers of either syngeneic or allogeneic lymphocytes clustered around macrophages from dusted rats compared with control rats. After 22 hr culture. Sustained lymphocyte binding was observed only with co-cultures containing syngeneic lymphocytes and macrophages from dusted rats. A considerable number of allogeneic lymphocytes or lymphocytes from control rats were still attached to the dusted macrophages after 22 hr, however, and a similar effect could be produced by pretreatment of control macrophages with NaIO4.

Sequential treatment of control macrophages with periodate and L-cysteine reduced the number of bound lymphocytes to those bound to untreated control macrophages. Pretreatment of the dusted macrophages with L-cysteine produced a sharp decline in the number of control lymphocytes or allogeneic dusted lymphocytes bound to the treated macrophage after 22 hr. Sustained lymphocyte-macrophage binding in co-cultures containing syngeneic lymphocytes and macrophages from dusted rats was not abolished by L-cysteine.

Thus, two different mechanisms could explain the prolonged binding of dusted alveolar macrophages with T lymphocytes. One mechanism observed in co-cultures of dusted macrophages with control lymphocytes or allogeneic dusted lymphocytes was analogous to that produced by treatment of control macrophages with NaIO4, and could be abolished by L-cysteine. It is postulated that asbestos exposure may produce an effect on the alveolar macrophage membrane similar to peroxidation. Another mechanism was demonstrated in co-cultures of syngeneic dusted macrophages and dusted lymphocytes where the sustained clustering observed after 22 hr was not abolished by L-cysteine. This finding is consistent with a specific immune recognition of an asbestos-related macrophage-associated antigen. Both these alterations would be due to the inhalation of crocidolite asbestos over a prolonged period.
Murine Cell Surface Immunoglobulin: Two Forms of δ-Heavy Chain
Authors: Mescher Matthew F. , Pollock Roberta R.
Published: December 2022

The heavy chain of isolated murine cell surface IgD is present in two forms, separable by electrophoresis on SDS-polyacrylamide gradient gels. Both forms of δ-heavy chain are present on the surface of intact spleen cells and have apparent m.w. of approximately 70,000 (δ1) and 68,000 (δ2). Treatment of surface IgD with neuraminidase before isolation results in a single IgD heavy chain band on SDS gels having an apparent m.w. of 65,000, indicating that δ1 and δ2 differ in sialic acid content. Delta2 is removed from the cell surface by papain more readily than δ1, suggesting a possible functional significance for the two forms.
Binding Constants of NZB Myeloma Antidextrans for Dextrans and Isomaltose Oligosaccharides Determined by Affinity Electrophoresis
Authors: Sugii Shunji , Takeo Kazusuke , Kabat Elvin A.
Published: December 2022

Association constants of dextrans (Ka) and oligosaccharides (Kia) from NZB myeloma antidextrans (PC3858 and PC3936) were studied by affinity electrophoresis. With linear dextrans or with those with a low degree of branching, Ka ranged from 2.7 × 103 to 5.4 × 104 ml/g for PC3858 and from 1.3 × 104 to 2.6 × 105 ml/g for PC3936. Completely linear α-(1 → 6)-linked dextrans, LD7 and D3, showed relatively high affinities for the two NZB antidextrans. With oligosaccharides, the Kia value increased as the number of α-(1 → 6)-linked glycosyl residues increased. Isomaltoheptaose (IM7) showed the highest Kia (1.9 × 104 M-1 for PC3858 and 1.63 × 104 M-1 for PC3936), whereas isomaltose (IM2) had the lowest Kia (2.36 × 102 M-1 for PC3858 and 1.32 × 102 M-1 for PC3936). Pullulan and glycogen showed very weak affinity for PC3936, but they did not react at all with PC3858. These findings indicate that NZB myeloma antidextrans, PC3858 and PC3936, are specific for internal chains of α-(1 → 6)-linked dextrans. Data on the precision with which Ka and Kia can be determined are presented.
The Pathogenesis of Sindbis Virus Infection in Athymic Nude Mice
Authors: Hirsch Robert L. , Griffin Diane E.
Published: December 2022

The pathogenesis of avirulent Sindbis virus infection was studied in normal and athymic nude mice. Marked differences in the immune response to infection were observed in nude and normal animals. In contrast to the intense mononuclear inflammatory cell response seen in the brains of normal mice 5 to 7 days after intracerebral injection of Sindbis virus, minimal inflammatory changes were observed in nude mouse brains. In addition, only IgM anti-Sindbis virus antibody was detected in nude mice whereas normal animals produced IgG as well. Despite these differences, the clinical outcome of the infection and the clearance of virus from the CNS was the same in both groups of mice. After footpad inoculation of virus, the lymphoproliferative responses of popliteal lymph node cells (LNC) from nude mice to Sindbis virus were meager or absent whereas normal LNC responded well (stimulation index = 38, 6 days after infection). Differences in the kinetics of the neutralizing antibody response to Sindbis virus were also observed. Although Ab production was not different through day 6, normal animals possessed 10-fold more anti-viral antibody 10 days after inoculation. It is concluded from these studies that mature T cells do not play a role in the clearance of Sindbis virus from the CNS of mice and that the early Ab response (IgM) and/or nonspecific defense mechanisms may be of greater importance in recovery from acute viral encephalitis.
20α Hydroxysteroid Dehydrogenase (20αSDH) Activity in New Zealand Mice T Lymphocytes and Bone Marrow Cells: Effect of Age, Sex, and Castration
Authors: Fuks Alejandro S. , Weinstein Yacob
Published: December 2022

The activity of 20α hydroxysteroid dehydrogenase (20αSDH), a T lymphocyte-associated enzyme, was measured in fetal liver, thymus, spleen, and bone marrow cells from NZB, NZW, and (NZB × NZW)F1 (B/W) mice. There was an age-dependent increase of 20αSDH activity in bone marrow cells, and a decrease in thymocytes and splenic T lymphocytes. Treatment with anti-θ and complement did not reduce the 20αSDH activity of bone marrow and fetal liver cells, but reduced the activity of spleen cells. PHA stimulates both 20αSDH activity and thymidine incorporation in splenic, bone marrow, and fetal liver lymphocytes. The results suggest that the enzyme in the bone marrow and fetal liver is located in pre-T lymphocytes. Enzymatic activity in bone marrow cells taken from female B/W mice (older than 7 months) was 40 to 50% lower than in male mice. Orchidectomy, but not ovariectomy, caused a significant decrease in thymocyte 20αSDH activity. Orchidectomy depressed and ovariectomy enhanced 20αSDH activity of bone marrow cells. The 20αSDH activity of fetal liver cells from B/W mice was twice as high as in either parent strain. No 20αSDH activity was found in fetal liver cells taken from BALB/C SJL or C57BL/6 mice.

A model is proposed to explain the age- and sex-related changes in 20αSDH activity of pre-T and T lymphocytes in healthy and pathologic conditions.