bluuebunny/crossref_metadata_2025 huggingface dataset:
Generation of antigen-specific suppressor cells in vitro in man.
Authors: Morimoto C , Reinherz E L , Todd R F , Distaso J A , Schlossman S F
Published: December 2022

The generation of human antigen-specific suppressor cells in vitro was examined in a primary anti-DNP antibody-forming system. It was found that specific T8 suppressor cells could be induced in vitro with high doses of antigen. The suppression obtained was macrophage dependent and carrier specific. KLH-specific T8 suppressor cells suppressed the anti-DNP antibody response induced by DNP-KLH but not by DNP-FGG. Similarly, T8 suppressor cells induced with FGG specifically suppress the anti-DNP response stimulated by DNP-FGG. These T8+ suppressor cells bear Ia molecules arising as a consequence of activation. This in vitro antigen-specific system should be useful for the further dissection of human immunoregulatory circuits.
Induction of antigen-specific antibody response in human peripheral blood lymphocytes in vitro by a dog kidney cell vaccine against rabies virus (DKCV).
Authors: Uytdehaag F G , Osterhaus A D , Loggen H G , Bakker R H , van Asten J A , Kreeftenberg J G , van der Marel P , van Steenis B
Published: December 2022

In the present report an in vitro method for obtaining a secondary human antibody response to a dog kidney cell vaccine against rabies virus (DKCV) is described. Cultures of peripheral blood mononuclear cells from normal rabies-immune and nonimmune donors were stimulated in vitro by DKCV. The production of virus-specific antibody in supernatant fluids was monitored by ELISA. Antibody was produced by lymphocytes from rabies-immune individuals, whereas those of nonimmune subjects consistently failed to produce anti-rabies antibodies after in vitro stimulation with DKCV. The generation of the anti-rabies virus antibody response of lymphocytes stimulated with DKCV was shown to be an antigen-dependent, as well as an antigen-specific process. Optimal antigen-specific responses were observed at relatively low concentrations of antigen (10(-1) to 10(-2) micrograms/culture). At increasing concentrations of antigen in culture (greater than 1 microgram/culture), the anti-rabies virus response was suppressed. Antibody produced upon stimulation was capable of neutralizing rabies virus. The response to rabies virus requires T cell help because lymphocytes depleted of SE rosetting cells did not respond to an antigenic stimulus. Studies in which the same individuals were followed over time showed a sequential development of circulating B cell subsets. The system may provide a model for the study of human B cell differentiation in vivo and in vitro and may be valuable for testing the potency of rabies vaccines in vitro.
Anti-idiotype as a probe in the analysis of autoimmune tubulointerstitial nephritis in the Brown Norway rat.
Authors: Zanetti M , Mampaso F , Wilson C B
Published: December 2022

Rabbit antibodies specific for the idiotype (Id) of autoantibodies to tubular basement membrane (TBM) eluted from kidneys of Brown Norway rats with tubulointerstitial nephritis (TIN) were used to analyze the immune response to TBM antigens at the humoral and cellular levels. These antibodies appeared to recognize Id determinants associated with the antigen combining site on the anti-TBM Id as well as on splenic lymphocytes. However, efforts to detect Id-positive cells in the interstitial infiltrates of kidneys with TIN failed. In contrast, in vivo injection of anti-Id serum before immunization with TBM resulted in a) significant selective suppression of antibodies to the autologous collagenase-solubilized TBM moiety but not to antigenic determinants of the intact TBM nor to those of heterologous TBM, and b) a corresponding decrease of TIN. These results suggest that anti-Id antibodies of this type can be useful as a probe for further dissecting the pathogenetic mechanisms underlying these complex autoimmune responses.
Comparative analysis of the influences of human gamma, alpha and beta interferons on human multipotential (CFU-GEMM), erythroid (BFU-E) and granulocyte-macrophage (CFU-GM) progenitor cells.
Authors: Broxmeyer H E , Lu L , Platzer E , Feit C , Juliano L , Rubin B Y
Published: December 2022

Preparations of human interferon (HuIFN) immune (gamma) (2 X 10(7) units/mg protein), HuIFN leukocyte (alpha) (1.4 X 10(8) units/mg protein) and HuIFN fibroblasts (beta) (10(6) U/mg protein) were assessed for their influence on colony formation of human hematopoietic progenitor cells: colony forming unit-granulocyte, erythroid, macrophage, megakaryocyte (CFU-GEMM), burst forming unit-erythroid (BFU-E), day 7 colony forming unit granulocyte-macrophage (CFU-GM) and day 14 CFU-GM. Colony formation by CFU-GEMM and BFU-E was suppressed equally by the three preparations of HuIFN, but colony formation by CFU-GM was suppressed differentially. CFU-GM were, on the whole, more responsive to HuIFN gamma than HuIFN alpha, and HuIFN beta was least effective. HuIFN alpha, but not HuIFN gamma or HuIFN beta, suppressed colony formation from CFU-GM without also suppressing the total number of colonies plus clusters. This was due to an increase in the numbers of clusters formed in the presence of HuIFN alpha. The suppressive influence on colonies from CFU-GM by the preparations of HuIFN and the enhancement of clusters by HuIFN alpha was apparently equal for colonies and clusters of neutrophils, eosinophils, macrophages and neutrophils plus macrophages. The suppressive effects of HuIFN gamma were inactivated by a monoclonal antibody to HuIFN gamma and the suppressive and enhancing effects of HuIFN alpha were inactivated with a heteroantiserum to HuIFN alpha. Depletion of monocytes, T lymphocytes and B lymphocytes from the target bone marrow cells had no influence on the effects of the preparations of HuIFN. These results demonstrate that the effects of HuIFN gamma and HuIFN alpha are due to the HuIFN themselves and that these actions on the hematopoietic progenitor cells are probably not mediated through monocytes and/or lymphocytes.
Human B cell differentiation. IV. Effect of monoclonal anti-immunoglobulin M and D antibodies on B cell proliferation and differentiation induced by T cell factors.
Authors: Kuritani T , Cooper M D
Published: December 2022

Monoclonal anti-mu and anti-delta antibodies and mixed lymphocyte reaction-derived T cell factors (MLR-TF) were examined alone and in combination for their effects on proliferation and differentiation of human B cells. MLR-TF induced proliferation and subsequent plasma cell differentiation of blood B cells without additional stimulation. The monoclonal anti-mu and anti-delta antibodies alone did not induce proliferation, but each was capable of augmenting B cell proliferation induced by MLR-TF. In contrast, the anti-mu antibody inhibited the MLR-TF induction of IgM plasma cell differentiation but did not affect the differentiation of IgG and IgA plasma cells. The anti-delta antibody had no effect on MLR-TF-induced plasma cell differentiation. Studies using density gradient separation of B cell subpopulations suggest that MLR-TF induce low-density B cells to proliferate and differentiate into plasma cells but that by themselves MLR-TF have little effect on B cells of relatively high density. The latter subpopulation of small resting B cells responded with proliferation to MLR-TF when combined with either the anti-mu or the anti-delta antibody, but these stimuli were insufficient for induction of terminal plasma cell differentiation.
Identification and characterization of gp TFA-1, a guinea pig T cell surface antigen associated with T cell function.
Authors: Burger R , Reske K , Mauer U , von Steldern D , Husmann M
Published: December 2022

Monoclonal antibodies (Ab) were produced that specifically recognized guinea pig T cells. FACS analysis revealed that Ab 188 bound to the majority of peripheral T lymphocytes of strain 2 and strain 13 guinea pigs and to a minor population of thymocytes. It failed to react with the Ia-bearing guinea pig B cell leukemia line EN-L2C, with macrophages, bone marrow cells, erythrocytes, or thrombocytes. Treatment of T cells with Ab 188 and complement prevented T cell activation. Culturing primed T cells with antigen- or mitogen-pulsed syngeneic or with allogeneic macrophages in the continuous presence of Ab 188 produced a marked, dose-dependent inhibition of T cell proliferation. The antigen defined by Ab 188 was therefore designated guinea pig T lymphocyte function-associated antigen-1, gp TFA-1. The magnitude of inhibition by Ab 188 varied between 65 and 85% whereas three other antibodies to guinea pig T cells had no inhibitory effect on T cell proliferation. Time course experiments revealed that gp TFA-1 is critically involved in an early phase of T cell activation. Maximal inhibition was achieved only if the antibody was present from the beginning of the cell culture; the addition of antibody after 24 hr of culture no longer had an inhibitory effect. Ab 188 did not induce T cell mitogenesis. Two-dimensional analysis (one-dimensional, IEF; two-dimensional, SDS-PAGE) of immunoprecipitates obtained from NP40 lysates of [35S]methionine-labeled T cell blasts indicated that a molecule was specifically precipitated that consisted of two noncovalently associated polypeptide chains with apparent m.w. of 43,000 and 38,000. Both subunits displayed extensive charge heterogeneity focusing at an average isoelectric point of 5.0 and 6.5, respectively. The gp TFA-1 molecule exhibits striking similarities in its functional and structural properties to recently described clonotypically expressed T cell glycoproteins, which were shown to be involved in antigen recognition by T cells in the murine and human systems.
Lymphocyte recognition of lymph node high endothelium. V. Isolation of adhesion molecules from lysates of rat lymphocytes.
Authors: Chin Y H , Carey G D , Woodruff J J
Published: December 2022

Lymphocytes migrate from blood into lymph nodes (LN) of rats specifically at segments of venules lined by high endothelium (HEV). Investigation of lymphocyte surface molecules mediating this interaction has been carried out using an in vitro assay in which lymphocytes adhere selectively to HEV when overlaid onto sections of peripheral LN. Using this assay we have previously isolated a thoracic duct lymph component termed High Endothelial Binding Factor (HEBF), which is detected by its ability to block HEV sites where lymphocytes attach. In the current study, we present evidence that anti-HEBF antibody recognizes surface molecules of lymphocytes involved in adhesion to high endothelium. Rabbit antibody was produced against HEBF isolated from lymph by sequential ion exchange [diethylaminoethyl (DEAE)-cellulose and carboxymethyl (CM)-Sepharose] and Sephacryl S-200 gel filtration. Anti-HEBF F(ab')2 bound to 60 to 70% thoracic duct lymphocytes (TDL), spleen, and LN cells but to only 2 to 9% thymus and bone marrow cells (indirect immunofluorescence). In addition, biologically active HEBF was isolated from lymph and detergent lysates of lymphocytes by antibody affinity chromatography. Comparable amounts of the factor were isolated from lysates of TDL, LN, and spleen cells whereas little or no HEBF was detected in lysates of thymus or bone marrow cells or in serum. HEBF obtained from TDL appeared to be a glycoprotein because it was trypsin sensitive, bound to lentil lectin, and was eluted with alpha-methyl-D-mannoside. The finding that HEBF was not recovered from lysates of trypsinized TDL indicates that the activity was mediated by components expressed on cell surface membranes. Immunoprecipitation studies revealed that anti-HEBF antibody recognized radioiodinated surface membrane proteins of TDL and TDL-derived T cells and B cells that resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) autoradiography into three major bands with m.v. of 230,000, 210,000, and 92,000 (nonreduced) and one major band with m.w. of 70,000 and two minor bands with m.w. of 92,000 and 45,000 (reduced). These observations indicate that HEBF is a surface membrane component that is not class-specific. It is suggested that lymphocyte surface HEBF is composed of high endothelial adhesion molecules that mediate cell-cell interactions involved in entry of lymphocytes from blood into peripheral lymph nodes.
Selective inhibition of platelet activation by the amyloid P-component of serum.
Authors: Fiedel B A , Ku C S , Izzi J M , Gewurz H
Published: December 2022

One component of amyloid, protein AP, has a characteristic pentameric structure and is identical with a 9.5s serum alpha 1-globulin designated serum amyloid P-component or SAP. Another pentameric molecule, the acute-phase reactant C-reactive protein (CRP), shares major amino acid sequence homology with SAP although, in man, SAP is not an acute-phase reactant. Recently, we demonstrated that heat-aggregated CRP (H-CRP), like heat-aggregated IgG, activates platelets to reactions of aggregation, secretion, and generation of thromboxane A2. We report here that physiologic concentrations of SAP inhibit platelet aggregation stimulated by H-CRP. SAP must be present before platelet challenge with H-CRP to be effective. Native (unaggregated) CRP does not inhibit platelet activation induced by H-CRP, and the platelet inhibitory effect of SAP is restricted because platelet responses to each heat-aggregated IgG, acid-soluble collagen, DNA, ADP, and thrombin remain unaltered in the presence of SAP. Thus, human SAP seems to selectively modulate platelet reactivity to modified CRP, and as such to down-regulate at least one aspect of the biologic capacity of its acute-phase homologue.
Degradation of the heparin matrix of mast cell granules by cultured fibroblasts.
Authors: Atkins F M , Metcalfe D D
Published: December 2022

The ability of cultured rat fibroblasts to phagocytose rat peritoneal mast cell granules has been previously demonstrated by light and electron microscopy. To determine if the heparin matrix of ingested granules could be degraded by fibroblasts after phagocytosis, the heparin within peritoneal mast cells was labeled with [35S]sulfate in vivo. The 35S-labeled rat peritoneal mast cells were purified and their granules were isolated and shown to contain [35S]heparin proteoglycan. Incubation of [35S]heparin proteoglycan-containing granules with cultured rat fibroblasts revealed internalization of radioactivity by the fibroblasts over the first 24 hr consistent with phagocytosis of the granules by these fibroblasts. The [35S]heparin proteoglycan internalized by the fibroblasts was shown to decrease in size over 72 hr indicating that the fibroblasts were capable of degrading the heparin within the ingested granules. Degradation of [35S]heparin proteoglycan within the fibroblast was accompanied by the appearance of free [35S]sulfate in the extracellular compartment. Similar findings were obtained using cultured human fibroblasts. These data demonstrate for the first time that both rat and human fibroblasts are not only capable of ingesting mast cell granules but also of degrading mast cell granule heparin proteoglycan. This ingestion and degradation of mast cell granules by fibroblasts may represent an important mechanism in the regulation of the biologic expression of heparin and other granule-associated mediators in immediate hypersensitivity reactions.
Studies on the induction and expression of T cell-mediated immunity. XIV. Antigen-nonspecific oxidation-dependent cellular cytotoxicity (ODCC) mediated by sodium periodate oxidation of cytotoxic T lymphocytes.
Authors: Fan J , Bonavida B
Published: December 2022

Alloimmune murine thymus-derived cytotoxic lymphocytes (CTL) generated in vivo or in vitro are shown to lyse antigen-nonspecific target cells (tumor cells, Con A, and LPS blasts) following treatment of CTL with an oxidizing agent, sodium periodate (NaIO4). It has been shown that NaIO4 oxidizes terminal sialic acid residues of cell surface macromolecules. The presence of reactive aldehyde groups, generated by NaIO4 modification, is required for the expression of antigen-nonspecific cytotoxicity because treatment of modified cells with a reducing agent such as potassium borohydride (KBH4) resulted in the abrogation of cytotoxicity. However, KBH4 treatment of unmodified or NaIO4-modified CTL has no effect on antigen-specific cytotoxicity. The modification of CTL by NaIO4 is sufficient to lead to the formation of lymphocyte-target cell conjugates and lysis of bound targets. Monoclonal antibodies directed against the Lyt-2 antigens of CTL, but not Lyt-1 antigens, in the absence of complement inhibited the nonspecific cytotoxicity resulting from NaIO4 modification of effector lymphocytes. These findings suggest that the mere interaction with or perturbation of appropriate cell surface molecule(s) of effector lymphocytes such as Lyt antigens by receptor-ligand interaction in SCMC or by NaIO4 modification in ODCC may lead to the expression of cytotoxicity. The present studies demonstrate a functional role of surface carbohydrates on CTL in cell-to-cell recognition and interactions. Furthermore, the results suggest that target cell modification is not a requisite for recognition and lysis in an antigen-nonspecific cytotoxic system such as ODCC. However, partial blocking of ODCC by alloantibodies directed against the H-2 of unmodified target cells suggests that NaIO4-modified CTL recognize unrelated target H-2 antigens. The implication of these findings on the molecular mechanism of cell-mediated cytotoxicity is discussed.
Mechanisms of protective immunity against Schistosoma mansoni infection in mice vaccinated with irradiated cercariae. IV. Analysis of the role of IgE antibodies and mast cells.
Authors: Sher A , Correa-Oliveira R , Hieny S , Hussain R
Published: December 2022

Mice resistant to challenge infection with Schistosoma mansoni by vaccination with highly irradiated cercariae were examined for the presence of circulating IgE antibodies and peritoneal mast cells sensitized against schistosome antigens. Significant levels of SWAP- or CAP-specific IgE antibodies could not be detected by solid phase radioimmunoassay in the sera of C57BL/6 mice during the first 6 wk after vaccination. Similarly, heatlabile antibodies capable of passively sensitizing normal mast cells for degranulation in response to SWAP could not be identified in the same sera. In contrast, peritoneal mast cells harvested from C57BL/6 mice 2 wk or later after vaccination gave strong degranulation responses when challenged with SWAP or CAP. Thus, vaccination with irradiated cercariae induces an unusual form of immediate-type hypersensitivity in which mast cells become sensitized in the absence of detectable circulating IgE antibodies. Mice deficient in mast cells (W/Wv mutant strain) were observed to develop the same resistance to challenge infection after vaccination with irradiated cercariae as nondeficient littermates. Similarly, vaccinated SJL/J mice were found to mount an extremely weak IgE response as measured by mast cell degranulation yet displayed the same level of resistance to challenge infection as other inbred mice developing potent mast cell responses. These findings argue that IgE antibodies and mast cells are not essential components in the effector mechanism of irradiated vaccine-induced immunity against schistosome infection.
Interleukin 2 deficiency in murine Leishmaniasis donovani and its relationship to depressed spleen cell responses to phytohemagglutinin.
Authors: Reiner N E , Finke J H
Published: December 2022

This study examined the kinetics and mechanisms of depressed spleen cell responses to phytohemagglutinin (PHA) that occur during Leishmania donovani infection of BALB/c mice. In co-culture experiments, neither spleen cells from infected animals nor parasite-infected macrophages suppressed PHA responses of normal spleen cells. In addition, parasite-mediated suppression of PHA-stimulated spleen cell proliferation could not be demonstrated. Mice with 2 wk of infection did manifest an impairment in spleen cell production of interleukin 2 (IL 2) and by 8 wk IL 2 activity in supernatants from these cells was reduced by approximately 95%. This finding was not explained by an alteration in the kinetics of IL 2 production. Furthermore, diminished IL 2 activity in supernatants of PHA-activated spleen cells from infected animals was not caused by suppressive factors in these fluids as shown by their inability to suppress IL 2 stimulation of IL 2-dependent T cells. When spleen cells from mice with 8 wk of infection were cultured with PHA and supplemented with exogenous IL 2, there was an approximately 48% increase in mitogenesis. These data indicate that abnormal PHA-induced spleen cell activation in BALB/c mice with L. donovani infection is associated with impaired production of IL 2. In addition, the observation that supplementation of spleen cells from infected mice with IL 2 resulted in partial reconstitution of the PHA response is consistent with a defect in IL 2 responsiveness.
Lack of general immunosuppression during visceral Leishmania tropica infection in BALB/c mice: augmented antibody response to thymus-independent antigens and polyclonal activation.
Authors: Colle J H , Truffa-Bachi P , Chedid L , Modabber F
Published: December 2022

Leishmania tropica causes a lethal visceral disease in highly susceptible BALB/c mice, with many immunopathologic features resembling those in human kala-azar. The responses to thymus-independent antigens of Type 1 and 2 (TI-1, TI-2) were compared in infected mice of susceptible BALB/c and resistant C57BL/6 strains at various times after infection. The infected BALB/c mice had an augmented response to both types of antigens at 45 days after infection. Later (day 76), the response to trinitrophenylated lipopolysaccharide (TNP-LPS, a TI-1 antigen) was diminished but that to dinitrophenylated Ficoll (DNP-Ficoll, a TI-2 antigen) remained statistically above the response of uninfected mice. The response of the resistant strain to either antigen was not modified as a result of the infection. Both strains showed significant polyclonal activation, which was considerably greater in the BALB/c than in the C57BL/6 mice. The observations presented here are in contrast to the widely held belief that a generalized nonspecific immunosuppression occurs in L. tropica infected BALB/c mice.
Effects of neonatal spleen cell products on the multiplication of toxoplasma in rat peritoneal macrophages.
Authors: Sakurai H , Igarashi I , Omata Y , Saito A , Suzuki N
Published: December 2022

We studied the resistance of newborn rats to infection with T. gondii. Neonates of chronically infected rats were significantly more resistant to infection with Toxoplasma than were the newborn of normal, uninfected rats. Neonatal athymic rats (nu/nu) from immune mothers were susceptible to infection, whereas normal littermates (nu/+) were very resistant. Rat peritoneal macrophage monolayers did not exhibit enhanced toxoplasmacidal activity after the addition of immune serum to the culture medium. However, inhibition of Toxoplasma multiplication was observed when the parasites were exposed to immune serum just before the infection of macrophages. The effects of spleen cell products from neonates of chronically infected mothers on the multiplication of Toxoplasma in rat peritoneal macrophages were examined. A significant inhibition of Toxoplasma multiplication occurred in macrophages incubated with the supernatants of spleen cell cultures obtained from immune newborn rats, with added lysate antigen. The ability to be induced to make an inhibitory supernatant disappeared by the third week after birth. Also examined were supernatants of spleen cells sensitized in vitro by incubation with a Toxoplasma lysate antigen; when these were added to macrophage monolayers, inhibition of Toxoplasma multiplication was observed only if the antigen was accompanied by supernatant of immune spleen cells.
Natural killer cell depletion enhances virus synthesis and virus-induced hepatitis in vivo.
Authors: Bukowski J F , Woda B A , Habu S , Okumura K , Welsh R M
Published: December 2022

The role of natural killer (NK) cells in the natural resistance of mice to infections by several viruses was examined. Mice were specifically depleted of NK cells by i.v. injection of rabbit antiserum to asialo GM1, a neutral glycosphingolipid present at high concentrations on the surface of NK cells. Control mice were left untreated or were injected with normal rabbit serum. Four to 6 hr later, these mice were infected with lymphocytic choriomeningitis virus (LCMV), mouse hepatitis virus (MHV), murine cytomegalovirus (MCMV), or vaccinia virus. The mice were sacrificed 3 days post-infection and assayed for virus in liver and spleen, spleen NK cell activity, and plasma interferon (IFN). All mice treated with anti-asialo GM1 antibody had drastically reduced NK cell-mediated lysis. Correlating with NK cell depletion, these mice had significantly higher (up to 500-fold) titers of MCMV, MHV, or vaccinia virus in their livers and spleens as compared to control mice. NK cell-depleted MCMV and MHV-infected mice had higher levels of plasma IFN than controls, correlating with the higher virus titers. These NK cell-depleted, virus-infected mice had more extensive hepatitis, assayed by the number of inflammatory foci in their livers, as compared to control virus-infected mice; these foci were also larger and contained more degenerating liver cells than those in control mice. In contrast to the results obtained with MHV, MCMV, and vaccinia virus, NK cell depletion had no effect on virus titers in the early stages of acute LCMV infection or during persistent LCMV infection. Mice depleted of NK cells had similar amounts of LCMV in their spleens and similar plasma IFN levels. Because this antibody to asialo GM1 does not impair other detectable immunologic mechanisms, these data support the hypothesis that NK cells act as a natural resistance mechanism to a number of virus infections, but suggest that their relative importance may vary from virus to virus.
Increase in NK cell number and turnover rate during acute viral infection.
Authors: Biron C A , Turgiss L R , Welsh R M
Published: December 2022

NK cell turnover and division during infection of mice with lymphocytic choriomeningitis virus (LCMV) was examined. Treatments with hydroxyurea (HU), a drug specific for cells synthesizing DNA, had no effect on control spleen NK cells, but significantly reduced NK cell-mediated lysis in LCMV-infected mice. When HU was administered at 9 and 2 hr before spleen cell harvest, the augmented lysis mediated by NK cells isolated from LCMV-infected mice was only 40% of that mediated by NK cells isolated from LCMV-infected untreated mice. Spleen cells that had been pulse-labeled with [3H]thymidine were used in a single cell assay with autoradiography to detect lysis mediated by blast cells. HU eliminated blast-NK cells; 34% of activated NK cell killing was mediated by blast cells, whereas less than 12% of the reduced NK cell activity after HU treatments was blast cell-mediated. The NK cells activated in vivo had lower densities than control NK cells. Although the total cell yields from control spleens and spleens on day 3 post-infection with LCMV were similar, low density Percoll gradient fractions from the day 3 spleen cell preparations contained three times as many cells as equivalent fractions from control populations. The same low density fractions were enriched in NK cell activity and in blast-NK cells. Activated spleen cell populations also contained increased numbers of large granular lymphocytes. These data indicate that NK cells have increased turnover rates during viral infection, that this increase in turnover is accompanied by NK cell division, and that this results in increases in the numbers of NK cells.
Demonstration of Aleutian disease virus-specific lymphocyte response in mink with progressive Aleutian disease: comparison of sapphire and pastel mink infected with different virus strains.
Authors: Race R E , Bloom M E , Coe J E
Published: December 2022

Lymphocyte blastogenesis was used to study the antiviral lymphocyte response of sapphire (Aleutian) and pastel (nonAleutian) mink inoculated with Pullman or Utah 1 Aleutian disease virus (ADV). Both mink genotypes developed a virus-specific response when inoculated with Utah 1 ADV. In contrast, after inoculation of Pullman ADV, sapphire mink had a positive virus-specific response, whereas pastel mink did not. Response occurred late after infection (8 wk) and correlated with the development of progressive Aleutian disease (AD). The response to keyhole limpet hemocyanin (KLH) and concanavalin A (Con A) was also determined. Most mink of either genotype, inoculated with either virus strain, maintained an anti-KLH response during disease. Most mink also responded to Con A, although some exhibited suppressed Con A response late in the disease course. These results indicated that mink develop an anti-ADV lymphocyte response during progressive AD and are not immunosuppressed with regard to other antigens or mitogens.
An allotype linked gene that is associated with a negative or very low anti-phosphorylcholine response (PC) phenotype in wild mice (CNV).
Authors: Lieberman R , D'Hoostelaere L A , Humphrey W , Nishinarita S , Potter M
Published: December 2022

In contrast to most inbred and wild mice, a population of wild mice recently isolated from a farm in Centreville, MD, and designated CNV produced no anti-phosphorylcholine (PC) antibodies (less than 1 microgram/ml) in response to immunization with the PC antigen Streptococcus pneumoniae (R36A) and gave 9 to 36 micrograms/ml anti-PC response to PC-KLH at 14 days after immunization. When another carbohydrate antigen, namely, bacterial levan, was used, CNV mice all gave high antibody titers. When CNV (PC-) mice were bred to inbred C.B20 (PC+) mice, 82% of the F1 and 76% of the F2 hybrids were surprisingly non-responders (PC-), which suggested that PC- gene(s) of CNV origin dominated the response to these antigens. The 18% PC+ phenotype in the F1 hybrids indicated possible heterozygosity of the PC genes controlling the PC- response in the CNV mice. Genetic studies on CNV mouse No. 378 supported this possibility. Analysis of the F2 data strongly suggest that two genes determined the PC- response, one of which was closely linked to the Igh-C allotype locus (chromosome 12). Hypothetically, we propose that CNV mice have two genes that cooperate but that sometimes act independently to express the PC- phenotype. Surprisingly, when F1 mice giving PC- phenotypes were back-crossed to C.B20, very few mice (18%) were PC-. This indicated that the PC- determining genes of CNV origin were not able to dominate immune responses in the presence of a larger number of C.B20 genes. This kind of expression may be regulated by other factors, such as clonotype competition and clonal dominance.
Xenoantisera against a murine T cell tumor product cross-react with immunoglobulin Fab determinants.
Authors: Mackel A M , Craddock G R , Warr G W , DeLuca D , Marchalonis J J
Published: December 2022

Some murine monoclonal T lymphoma cells express a surface component that reacts with chicken antisera produced against the Fab fragment of normal mouse IgG. In the present study, we use a solid phase immunoadsorbent consisting of affinity-purified chicken anti-Fab coupled to Sepharose to isolate a product produced by the in vitro T cell line, WEHI-7.1. The affinity-purified T cell surface molecule (IgT) migrated on SDS-PAGE as a single band of approximately 65,000 daltons. The object of these studies was to produce xenoantisera against the purified T cell product cross-reactive with Ig determinants and to characterize the antisera. Rabbits immunized with this purified molecule produced antibodies that reacted with Fab fragments of polyclonal mouse IgG and with the myeloma proteins MOPC-104E and MOPC-41, as detected by enzyme-linked immunosorbent assay (ELISA). This binding was eliminated by adsorption of the antisera with normal polyclonal IgG; however, adsorption with fetuin did not significantly affect the reactivity of the antisera. Radioimmune precipitation assays revealed that the rabbit anti-IgT bound to normal murine spleen and thymus cells; this reactivity was abrogated by adsorption with insolubilized polyclonal IgG. Competition radioimmunoassays demonstrated that detergent extracts of the thymus and the spleen contained material that inhibited the precipitation of MOPC-41; nonlymphoid cells lacked such material. The rabbit anti-IgT serum blocked the binding of antigen by normal T cells; adsorption of the antiserum with polyclonal IgG-Sepharose abrogated this blocking capacity. A solid phase immunoadsorbent prepared from the IgG fraction of the rabbit anti-IgT isolated a single component from formic acid-solubilized mouse thymus. This molecule had an approximate mass of 65,000 to 70,000 daltons. The anti-IgT serum isolated surface IgM and IgD from lactoperoxidase-catalyzed radioiodinated B cells. The anti-IgT serum detected IgM and IgG in mouse serum with the use of immunoelectrophoresis. The anti-IgT immunoadsorbent isolated several components from normal mouse serum, that, when analyzed by SDS-PAGE under reducing conditions, revealed bands corresponding to mu-, gamma-, and light chains as well as components that migrated between mu- and gamma-chains, and another component with an approximate mass of 45,000 daltons. Our results with antibodies to a purified T cell product indicate that a surface component of normal T cells and certain monoclonal T cell tumor lines is serologically related to the Fab fragment of serum Ig and is implicated in the binding of antigen.
Immunotherapy of a murine B cell tumor with antibodies and F(ab')2 fragments against idiotypic determinants of its cell surface IgM.
Authors: Perek Y , Hurwitz E , Burowski D , Haimovich J
Published: December 2022

Anti-idiotype antibodies against the IgM of the 38C-13 murine B lymphocyte cell line and F(ab')2 fragments derived from them were checked for their therapeutic effect against the killing of mice by 38C-13 cells. Both intact molecules and the F(ab')2 fragments were effective in delaying or preventing the killing of mice by 38C-13 tumors when used in large amounts and by the appropriate routes.
Induction of T cell responses in nonresponder mice by abolishing suppression with monoclonal antibodies recognizing A region-controlled, T cell-specific determinants.
Authors: Ikezawa Z , Nonaka M , Abe R , Tada T , Nagy Z A , Klein J
Published: December 2022

Mouse strains carrying the kappa allele at loci A beta, A alpha, E beta, and E alpha are nonresponders to lactate dehydrogenase B (LDHB) and to allotypic determinants of IgG2a myeloma proteins (for example, UPC10 used in this study). The nonresponsiveness to these antigens is caused by T suppressor (Ts) cells that prevent antigen-primed T helper (Th) cells from proliferating. We demonstrate here that monoclonal antibodies specific for an A region-controlled molecule selectively expressed on T cells (A-T) are capable of inducing anti-LDHB and anti-UPC10 responses of primed T cells from nonresponder strains. A monoclonal anti-J antibody that cross-reacts with the A-T molecule also induces responsiveness, whereas another J-specific antibody that lacks this cross-reactivity fails to do so. The mechanism of response induction is blocking of the interaction between the Ts cell or its factor (TsF) and the target of suppression, the antigen-specific Lyt-1+2- (Th) cell. The blocking occurs at the level of the Ts cell and the TsF. The data indicate that Ts cells and TsF carry a unique, A region-controlled molecule that is not only functionally analogous but also serologically similar to the J molecule.
Self recognition of accessory cell Ia determinants is required for the in vitro generation of hapten-specific cytotoxic T lymphocyte responses.
Authors: Kruisbeek A M , Andrysiak P M , Singer A
Published: December 2022

The present study has addressed the involvement of Ia determinants in the in vitro generation of antigen-specific cytotoxic T lymphocyte (CTL) responses. It demonstrated that the in vitro generation of TNP-specific CTL responses strictly requires responder T cell recognition of self-Ia determinants expressed by accessory cells, and that this recognition could be specifically inhibited by monoclonal anti-Ia-antibodies. The generation of TNP-specific CTL responses was unaffected by the presence of anti-I-A antibodies or the absence of accessory cells when cultures were performed in the presence of an exogenous source of T helper cell factors, Con A SN. Thus, these results indicate that T helper cell recognition of self-Ia determinants expressed by accessory cells is required for the generation of TNP-specific CTL responses. These results preclude the possibility that accessory cells perform only immunologically nonspecific roles in the generation of hapten-specific CTL, but instead demonstrate that accessory cells function in such responses as Ia-bearing antigen-presenting cells for the activation of self-Ia-specific T cells.
The cellular mechanism of maintenance of neonatally induced tolerance to H-2 class I antigens.
Authors: McCarthy S A , Bach F H
Published: December 2022

We used limiting dilution analysis protocols to investigate the mechanism by which in vitro cytotoxic T lymphocyte (CTL) hyporeactivity is maintained in adult mice that had been neonatally tolerized to major histocompatibility complex-encoded antigens. Class I molecules, presented on donor cells having an H-2 K or D region haplotype difference from recipients, readily induce tolerogen-specific CTL hyporeactivity. All attempts to identify any in vitro effects of active suppressive cells operative in the maintenance of this hyporeactivity have been unsuccessful. We conclude that this cytotoxic deficiency is the consequence of in vivo mediated clonal inactivation of the precursors of tolerogen-specific CTL. A presentation and evaluation of the assumptions inherent in this conclusion are made. In contrast to class I molecules, class II molecules, presented on donor cells having an H-2 I region haplotype difference from recipients, are unable to induce tolerogen-specific CTL hyporeactivity, even when injected neonatally at high doses. This inability of class II molecules to induce CTL tolerance parallels the considerable difficulty of inducing helper T lymphocyte tolerance to class II molecules.
Allergic contact dermatitis induced by intratracheal administration of hapten.
Authors: Stein-Streilein J
Published: December 2022

It is known that the skin is the natural route for induction of allergic contact dermatitis (ACD). Because the lung is another potential portal of entry for sensitizing chemicals, studies were performed to evaluate immune responses to haptens deposited in the lung. As a result, a new method for inducing ACD was developed. Intratracheal (IT) inoculation of 2,4,6-trinitrobenzene sulfonic acid (TNBS) led to maximal ear swelling 24 hr after challenge on the ear with 2,4,6-trinitrochlorobenzene (TNCB) in carrier. This response was specific for immunizing hapten. Furthermore, it was equally possible to induce ACD by intratracheal inoculation of trinitrophenyl- (TNP) modified bronchoalveolar cells (BAC) or haptenated spleen cells. Adoptive transfer studies demonstrated that the hypersensitivity that resulted from IT inoculation of TNBS or TNP BAC could be transferred with cells. With a monoclonal anti-hamster Ia reagent, it was demonstrated that, like Langerhans cells, 80 to 90% of cells in the bronchial lavage fluid expressed Ia determinants on their membrane. These cells were morphologically indistinguishable from macrophages. Because an individual is capable of being sensitized when hapten is introduced by the pulmonary or epicutaneous routes, the possibility is raised that the alveolar macrophages in the lung possess a similar capability for antigen presentation of hapten in the induction of ACD as does the Langerhans cell.
Generation and characterization of monoclonal antibodies reactive with human B lymphocytes.
Authors: Mittler R S , Talle M A , Carpenter K , Rao P E , Goldstein G
Published: December 2022

Four novel monoclonal antibodies directed at determinants on human B lymphocytes were produced and characterized by flow cytometry using indirect immunofluorescence analysis. The antibodies, OKB1, OKB2, OKB4, and OKB7, define antigens not previously described on human B lymphocytes using B cell reactive monoclonal antibodies or standard B cell phenotypic markers. OKB4 [immunoglobulin M (IgM)] and OKB7 (IgG2a) were reactive with greater than 95% of all surface membrane immunoglobulin positive (Smlg+) lymphocytes and were restricted to the B cell lineage. OKB2 (IgG1) likewise reacted with greater than 95% of all Smlg+ lymphocytes; however, the antigen was also expressed on all granulocytes. In contrast to OKB2, OKB4, and OKB7, OKB1 reacted with a variable percentage of normal peripheral B cells (approximately 70 to 95%). Functional studies employing polyclonally activated B cells in a pokeweed mitogen (PWM) driven system and the reverse hemolytic plaque assay for the quantitation of Ig secreting plasma cells demonstrated that OKB1 and OKB2 inhibited the generation of plaque-forming cells (PFC) when added to cultures at day 0. OKB4 had a marginal inhibitory effect, whereas OKB7 consistently enhanced the PFC response. Immunoprecipitation studies with OKB1, OKB4, and OKB7 demonstrated that these antibodies precipitated antigens of approximately 168,000, 87,000, and 175,000 m.w., respectively. The OKB2 antigen is presently under study.
Selective inhibition of anti-nucleoside-specific antibody production by nucleoside-ricin A conjugate.
Authors: Morimoto C , Masuho Y , Borel Y , Steinberg A D , Schlossman S F
Published: December 2022

The effect of the preincubation of peripheral blood lymphocytes, from SLE patients, with nucleoside-ricin A conjugates on spontaneous A,G,C,T antibody production was examined. Enhanced spontaneous anti-nucleoside-specific antibody (anti-A,G,C,T antibody) production by SLE B cells was selectively inhibited by pretreatment in vitro with nucleosides conjugated to the ricin A chain. The selective suppression was demonstrated by the lack of suppression of the anti-DNP response or of polyclonal IgG production by pretreatment that did suppress anti-A,G,C,T production by the lymphocytes of eight patients with SLE. Furthermore, pretreatment of B cells, but not of T cells, with nucleoside-ricin A conjugates inhibited the A,G,C,T antibody response by these B cells. Thus, (A,G,C,T)-BGG-ricin A conjugates bind directly to the nucleoside-specific B cells via their antigen receptors. This demonstration of the selective elimination of B cells might have therapeutic applications in SLE.
Human bone marrow allograft recipients: production of, and responsiveness to, interleukin 2.
Authors: Warren H S , Atkinson K , Pembrey R G , Biggs J C
Published: December 2022

The production and utilization of interleukin 2 (IL 2) by peripheral blood mononuclear cells (PBMC) from 14 bone marrow allograft recipients was examined. PBMC from all patients were impaired in their ability to produce IL 2 when stimulated by phytohemagglutinin (PHA). On the average, the IL 2 activity produced by patients' PBMC was 8% of that from normal donors' PBMC. To examine the basis for this impaired T cell function in marrow recipients, the ability of a resting T lymphocyte population isolated from PBMC to respond to PHA and exogenously supplied IL 2 was analyzed. The proliferative response of resting T cells to PHA and IL 2, although low at early times post-transplant, reached near normal levels by 8 mo. Only two of 11 patients had normal numbers of precursor T cells that could respond. For all other patients the average number of precursor T cells was 10-fold lower than the average determined for normal donors. The impaired production of IL 2 by patients' PBMC may be due to this low precursor frequency. For some patients the rate and/or extent of clonal expansion of activated T cells appears to be greater than that of normal donors. The data suggest that the therapeutic application of IL 2 to such patients is unlikely to be successful in overcoming defects of T cell function before 8 mo post-marrow transplant.
Purified human monocyte subsets as effector cells in antibody-dependent cellular cytotoxicity (ADCC).
Authors: McCarley D L , Shah V O , Weiner R S
Published: December 2022

Human peripheral blood monocytes are heterogeneous with respect to their size and function. Two monocyte subsets were isolated by countercurrent centrifugal elutriation and were studied with respect to their ability to effect antibody-dependent cellular cytotoxicity (ADCC) and for the presence of Fc receptors on their surface. Both monocyte subsets display Fc surface receptors and are effectors of ADCC against sensitized human erythrocyte target cells. The demonstration of ADCC by monocyte effectors is dependent on their concentration in the incubation mixture. Dilution of monocytes below 10% by unlabeled and unsensitized erythrocytes or lymphocytes significantly suppresses ADCC, presumably by steric inhibition of effector and target contact.
The simultaneous antagonistic effects of a T cell hybridoma product on the growth and the maturation of activated lymphocytes.
Authors: Primi D
Published: December 2022

Lymphocyte maturation and growth are two antagonistic events; therefore, successful antibody synthesis accompanied by clonal expansion, as seen in most humoral immune responses, must be the result of a delicate quantitative balance between maturation and growth factors. On the basis of this hypothesis, it should be feasible to search for a T cell product that alters this equilibrium and favors either B cell growth or maturation. In an attempt to isolate T cell hybridomas producing these activities, we fused BW5147 thymoma cells with Con A-activated spleen cells. One of the hybridomas obtained constitutively produces a product (or products) that selectively inhibits mitogen-induced replication but not maturation of normal but not transformed B lymphocytes. More importantly, the same product(s) supports Ig synthesis but not growth of LPS blasts, which suggests that we are dealing with a B cell maturation factor. The effect of this supernatant can be completely abrogated by the B cell mitogen DxS. In addition, the proliferative response of B cells to this ligand is unaffected by the hybridoma product. The implications of our results for understanding the mechanism of B cell triggering are discussed.