bluuebunny/crossref_metadata_2025 huggingface dataset:
T cell responses to the major allergen from the house dust mite Dermatophagoides pteronyssinus, Antigen P1: comparison of patients with asthma, atopic dermatitis, and perennial rhinitis.
Authors: Rawle F C , Mitchell E B , Platts-Mills T A
Published: December 2022

Peripheral blood mononuclear cells (PBMC) from the majority of the allergic patients that we tested who were skin test-sensitive to the house dust mite Dermatophagoides pteronyssinus (D. pt.) (46/67) showed significant proliferation in response to the purified major allergen Antigen P1; PBMC from 14/15 nonsensitive controls showed no significant response. Optimal responses were seen with 10 micrograms Antigen P1/ml, but 21 of 43 patients tested showed significant proliferation at 0.01 microgram Antigen P1/ml. The results of three types of experiments showed that the responding cells were T cells, predominantly of the helper phenotype. First, purified T cells in the presence of irradiated or mitomycin C-treated antigen-presenting cells showed good proliferation to Antigen P1. Secondly, flow cytometry analysis showed a progressive increase in the percentage of viable cells bearing the Leu-3a marker--up to 88% by day 7--and demonstrated that the larger, blast-transformed cells bore this marker. Finally, interleukin 2 production was demonstrated in antigen-stimulated cultures at days 3 to 5, i.e., about 2 days before the peak of proliferation. When patients were grouped according to their disease symptoms, no clear differences were seen between the T cell responses of patients with asthma, eczema, or rhinitis, or with asthma and eczema, although patients with rhinitis alone tended to show weaker responses. Overall, there was a significant correlation between serum IgE antibody to Antigen P1 and T cell proliferation (rs = 0.579, p less than 0.001); however, excluding individuals with no IgE antibody, the quantitative correlation was poor (rs = 0.26, p less than 0.1). These results indicate that most patients showing immediate hypersensitivity to D. pt. have circulating T cells sensitized to Antigen P1. These sensitized T cells probably act as helper cells for antibody production, but may also play a role in the pathogenesis of allergic lesions and in the delayed or chronic symptoms of these allergic diseases.
Murine interstitial nephritis. II. The adoptive transfer of disease with immune T lymphocytes produces a phenotypically complex interstitial lesion.
Authors: Zakheim B , McCafferty E , Phillips S M , Clayman M , Neilson E G
Published: December 2022

The present studies demonstrate that immune Thy-1.2+, Lyt-1.2+ T lymphocytes harvested from SJL mice with anti-tubular basement membrane disease can adoptively transfer interstitial nephritis into naive recipients. The lesions produced after cell transfer do not occur immediately but rather take 4 to 6 wk to fully develop. Interstitial lesions can also be transferred to a lesser degree and over a longer period of time with immune serum containing anti-tubular basement membrane antibodies. The fully formed lesions that developed after the transfer of immune cells or serum were phenotypically characterized by cell-surface antibodies using immunofluorescence. T lymphocytes, natural killer cells, macrophages, and Ig+ cells were all well represented in both lesions. Natural killer cells, however, were slightly more prevalent in the lesions of mice receiving immune serum. These experiments demonstrate a potential role for both immune T lymphocytes and anti-tubular basement membrane antibodies in the development of interstitial nephritis in mice. Unlike guinea pigs and rats, it is only in mice that interstitial lesions can be adoptively transferred with immune T lymphocytes, and as such, this model should prove very useful in the additional dissection of cellular interactions and immunoregulatory events that formulate the final effector mechanisms of disease expression.
Interleukin 1 activity in normal human urine.
Authors: Kimball E S , Pickeral S F , Oppenheim J J , Rossio J L
Published: December 2022

Urine concentrates from normal individuals were shown to contain interleukin 1 (IL 1)-like activity when tested directly on human dermal fibroblasts and on C3H/HeJ mouse thymocytes in the presence of 1 microgram/ml phytohemagglutinin. Seventy-five percent of the urine samples tested, however, demonstrated the presence of a specific inhibitor of IL 1-promoted thymocyte proliferation. This inhibitor did not affect IL 2-promoted proliferation of mouse thymocytes or CT-6 cells or IL 1-promoted proliferation of human dermal fibroblasts. After gel filtration of the urine concentrates, even those samples that were inhibitory yielded fractions containing both thymocyte and fibroblast proliferative activity. The approximate m.w. of these activities were 75,000 and 15,000. In addition, two peaks of low m.w. thymocyte proliferative activity were noted at 4000 and 2000. The 2000 pool, but not the 4000 pool, also contained fibroblast proliferative activity.
Release of discrete interferons by cytotoxic T lymphocytes in response to immune and nonimmune stimuli.
Authors: Pasternack M S , Bevan M J , Klein J R
Published: December 2022

We have examined the potential of murine cytotoxic T lymphocytes (CTL) to synthesize both IFN-gamma and IFN-alpha, beta by using three cloned antigen-reactive CTL lines. These lines, which produce IFN-gamma in an antigen-specific manner or after stimulation with mitogen, were also found to produce IFN-alpha, beta upon nonspecific contact with Newcastle disease virus. The requirement for viral integrity in this process has been examined. Virus infectivity was not necessary to stimulate the release of IFN-alpha, beta from CTL, but acid-inactivation of virus abrogated its IFN-inducing property.
Recognition of H-2Kb mutant target cells by Moloney virus-specific cytotoxic T lymphocytes from bm13 (H-2Db-mutant) mice. II. Relationship of Kbm3 and Kbm11 in restriction specificities and allodeterminants.
Authors: Stukart M J , Boes J , Melief C J
Published: December 2022

We have studied the effect of mutations in H-2Kb on recognition of Moloney virus antigens by Kb-restricted Moloney virus-specific cytotoxic T lymphocytes (CTL) generated from H-2Db-mutant B6.C-H- 2bm13 ( bm13 ) mice. On the basis of the bm13 virus-specific recognition pattern of a series of Kb-mutant virus-infected (V+) cells, these Kb mutants could be divided into two groups that either shared the relevant H-2K restriction specificities with H-2Kb ( bm3 , bm5 , bm6 , and bm11 ) or lacked them completely (bm1) and/or almost completely ( bm8 ). In the study presented in this report, we concentrated on the alloreactive CTL included in the repertoire of bm13 Moloney-specific CTL. These CTL lysed noninfected (V-) allogeneic target cells of many H-2 types, including all Kb-mutant cells tested (bm1, bm3 , bm5 , bm6 , bm8 , and bm11 ). Recognition of V- cells of two Kb mutants, bm3 and bm11 , which share a common amino acid substitution at position 77 of the H-2K molecule with an additional change at position 89 in bm3 , was compared with recognition of the other Kb-mutant V- target cells in cold target inhibition studies. These studies showed that bm3 and bm11 cells share a determinant or determinants on the H-2K molecule recognized by alloreactive CTL among the Moloney virus-specific CTL population not present on the other Kb-mutant cells; lysis of both target cells was inhibited only by bm3 and bm11 V- cells. On the other hand, lysis of bm3 V+ target cells by bm13 virus-specific CTL was inhibited by all Kb-mutant V+ cells ( bm3 , bm5 , bm6 , and bm11 ) that were killed virus specifically by bm13 CTL. Thus, common determinants on the H-2K molecule of these Kb-mutant V+ cells are recognized by Kb-restricted Moloney virus-specific CTL. From the difference in inhibition pattern of the lytic reaction against bm3 V+ vs bm3 V- cells, we conclude that determinants on the same H-2K molecule recognized by Moloney virus-specific CTL and allocross -reactive effector cells included in the Moloney virus-specific population are not identical.
Isolation and characterization of a human T lymphocyte-associated glycoprotein (gp40).
Authors: Sutherland D R , Rudd C E , Greaves M F
Published: December 2022

The biosynthetic and structural characteristics of the human thymocyte/T cell antigen defined by the monoclonal antibody WT1 have been studied. WT1 identified a monomeric cell surface glycoprotein of Mr = 40,000 ( gp40 ). Cross-absorption experiments and two-dimensional gel analyses indicate that WT1 and another monoclonal antibody, 3A1, react with the same structure. This glycoprotein was asymmetrically inserted into the rough endoplasmic reticulum as a transmembrane structure. At this stage, the polypeptide chain possessed two N-linked, "high-mannose" type glycans; these were subsequently processed into endo-H-insensitive, complex oligosaccharides during intracellular transport to the cell surface. Inhibition of N-linked glycosylation with tunicamycin failed to block the processing of the nonglycosylated Mr = 29,000 polypeptide to a glycoprotein of Mr = 33,000. Cleavage of the mature Mr = 40,000 form with endo-F yielded a similar Mr = 33,000 product. The kinetics of synthesis of the Mr = 33,000 intermediate in conjunction with gal-NAc oligosaccharidase digestion indicated the presence of O-linked glycans in the mature cell surface WT1 antigen. The fully processed cell surface form of the polypeptide also contains covalently associated fatty acid, and was labeled by 32P phosphate, the predominantly labeled phosphoamino acid being phosphoserine. We also demonstrate biochemically that the reactivity of WT1 with cells from a few patients with acute myeloid leukemia reflects genuine expression of the gp40 structure on myeloid cells.
Induction of minor alloantigen-specific T cell subsets in vivo: recognition of processed antigen by helper but not by cytotoxic T cell precursors.
Authors: Czitrom A A , Gascoigne N R , Edwards S , Waterfield D J
Published: December 2022

Primary cytotoxic T lymphocyte (CTL) and helper T (Th) cell responses were generated from regional lymph nodes of heterozygous mice sensitized in the footpad with spleen cells of parental H-2 genotype but differing for multiple minor alloantigens of the B10 background. Cytotoxic and helper responses were read after a short in vitro culture period, which did not influence the restriction specificities of in vivo sensitized T cells. We show that under these conditions, Th cell responses appear unrestricted, whereas CTL responses are restricted to the immunizing parental H-2 genotype. Furthermore, we present evidence that these two functions are mediated by distinct T cell subsets and that the in vivo induction of Lyt-2- Th cells has the appearance of being unrestricted whereas that of Lyt-2+ CTL is restricted by H-2. These findings suggest that in a primary immune response, in vivo precursors of Th cells recognize processed antigen on F1 antigen-presenting cells (clones restricted to each parental H-2 genotype are cross-sensitized) under conditions in which those of CTL only respond directly to foreign minor alloantigens on the injected stimulator cells (clones restricted to parental H-2 genotypes are not cross-immunized). This dichotomy is taken to suggest the possibility that processing of cell membrane-bound antigens is controlled at their association step with products of the major histocompatibility complex.
MALA-1: a surface antigen expressed on activated murine T and B lymphocytes.
Authors: Takei F
Published: December 2022

Detergent-solubilized plasma membranes of Con A-activated mouse spleen cells were absorbed with Sepharose-coupled rat antibodies against resting mouse lymphocytes. The unbound fraction was used to immunize a rat, and the immune spleen cells were fused with the rat myeloma Y3 . All seven rat monoclonal antibodies produced in this way strongly reacted with mitogen-activated spleen cells but only weakly or insignificantly with unstimulated spleen cells. One of the antibodies, YE3 /19.1, was studied in detail. The antibody strongly reacted with Con A- or LPS-stimulated spleen cells, but not significantly with normal adult thymocytes, spleen cells, or bone marrow cells. Unlike the transferrin receptor, which is expressed on virtually all dividing cells, the antigen defined by YE3 /19.1 was not detected on erythroblast-enriched populations or some non-T, non-B cell lines. Therefore, the antigen, termed MALA-1, seems to be specific for activated murine lymphocytes of the T and B cell lineages. Over 25% of normal adult lymph node cells were also found to express the antigen, although the antigen densities on lymph node cells were lower than those on mitogen-stimulated spleen cells. Kinetic studies of the expression of MALA-1 and the transferrin receptor on Con A-activated spleen T cells showed that both antigens are detectable within 24 hr of Con A stimulation. Although the density of the transferrin receptor on Con A blasts declined as the cell proliferation ceased, MALA-1 expression persisted. Immunoprecipitation of MALA-1 from surface-iodinated Con A blasts revealed its m.w. to be approximately 14,000 to 18,000.
Antigenic regions defined by monoclonal antibodies correspond to structural domains of avian lysozyme.
Authors: Smith-Gill S J , Lavoie T B , Mainhart C R
Published: December 2022

The epitopes recognized by six new BALB/c hybridomas specific for the protein antigen hen egg-white lysozyme (HEL) were mapped in detail. Although fine specificities of all the antibodies were distinct, many of the epitopes overlap in complex patterns. The antibodies could be grouped into three complementation groups, one of which also included the previously characterized HyHEL -5 which is specific for Arg68 . Complex interactions were observed among the antibodies, both among and within complementation groups, including nonreciprocal competition and enhanced binding. Two of the complementation groups mapped near the catalytic site in a new antigenic region. The antibodies HyHEL -8 and HyHEL -10 had very similar and over-lapping specificities, and may recognize very closely related epitopes. The results suggest that the epitopes may form a continuous antigenic surface, and that antigenic regions correspond to structural domains defined by the tertiary structure of HEL.
Activation of the alternative complement pathway by isolated human glomerular basement membrane.
Authors: Williams J D , Czop J K , Abrahamson D R , Davies M , Austen K F
Published: December 2022

The capacity of isolated human glomerular basement membrane (GBM) to initiate surface activation of the human alternative complement pathway was defined by the deposition of C3b under circumstances in which the classical complement pathway was inoperative. The deposition of C3b from normal or C2-deficient serum was time- and magnesium-dependent, implying a role for the alternative pathway. Normal human serum rendered deficient in D did not sustain C3b deposition until its reconstitution with D, indicating an absolute requirement for a protein unique to the alternative pathway and essential to the cleavage activation of the C3 amplification convertase of that pathway. The capacity of the excess control proteins H and I to prevent C3b deposition onto GBM incubated in C2-deficient serum provided further evidence for the direct activation of the alternative pathway in this system. The use of radiolabeled monoclonal antibody to localize the deposited C3b afforded specificity and quantitation of about 100 ng of C3b/mg of GBM. Immunohistochemical analysis with a monoclonal antibody to detect C3b demonstrated its deposition to be confined to the epithelial surface of the GBM.
Genetic polymorphism of the sixth component (C6) of rat complement.
Authors: Granados J , Cramer D V , Caputo J B , Marcus D , Alper C A
Published: December 2022

The complement protein C6 has been shown to be genetically polymorphic in the rat. Isoelectric focusing of plasma samples from 19 inbred strains demonstrated two electrophoretically distinguishable migration patterns, each consisting of three bands. Breeding studies with the use of the BN and DA strains showed that the C6 patterns were inherited in a manner consistent with the co-dominant autosomal expression of two alleles (C6 A and C6 B). The distribution of the C6 alleles in a backcross mating was compared with eight independently segregating marker genes: RT1.A, RT2, Gdc -1, Igk-1, Hbb, Svp-1, Fh-1, and Es-6. There was no detectable linkage between C6 and any of these eight loci.
Amino acid and nucleotide sequences of variable regions of mouse immunoglobulin light chains of the lambda 3-subtype.
Authors: Reilly E B , Reilly R M , Breyer R M , Sauer R T , Eisen H N
Published: December 2022

To learn about the V lambda gene segments that are expressed in lambda 3 light chains, the most recently identified lambda-chain subtype in inbred mice, we determined partial amino acid sequences of the V regions of two of these chains, L5-8 and Lc49 . The partial sequences were extended by establishing the complete V region sequence of cDNA for the lambda-chain mRNA from the hybridoma (RZ 5-8) and the myeloma ( CBPC -49) that produce these chains. The primer extension method used to sequence the cDNA is described in detail, because the same primer (a synthetic heptadecadeoxynucleotide ) can be used for sequencing cDNA for lambda-chains of all three subtypes of inbred mice and probably for lambda-chains from some other vertebrate species as well. The results confirm earlier preliminary findings that for both chains the V region is encoded by the V lambda 1 and J lambda 3 gene segments. The unmutated germ-line sequences of these gene segments are present in both chains, but the two chains, nevertheless, differ at codon 97, the V lambda-J lambda boundary. A T/G difference in the third position of this codon resulted in a codon for histidine (CAT) in one chain (L5-8) and for glutamine (CAG) in the other chain ( Lc49 ). This difference can be accounted for by variation in the site of V lambda 1-J lambda 3 recombination. Though the V region amino acid sequences of L5-8 and Lc49 differ only by the His/Gln substitution at position 97, the two chains have been shown (manuscript in preparation) to differ in their ability to form an effective combining site for the 2,4-dinitrophenyl group.
Overproduction of immunoglobulin mRNA by a murine myeloma MOPC 315 variant cell line.
Authors: Ponte P A , Dean M , Pepe V H , Sonenshein G E
Published: December 2022

The mouse myeloma MOPC 315 cell line synthesizes and secretes IgA (lambda 2) immunoglobulin. A spontaneously arising variant of the MOPC 315 line, which had been isolated as apparently oversecreting IgA protein, has been characterized. The variant line has been shown to synthesize and secrete increased levels of heavy chain, light chain, and J chain polypeptide compared to the parental wild-type cells from which it was isolated. The steady-state levels of cytoplasmic mRNA for these polypeptides are increased commensurately in the over-producing line. For the heavy chain, enhanced transcription, and possibly increased gene dosage, appear to be involved. The increased levels of the three individual immunoglobulin polypeptide chains suggest that the variant line displays a coordinate regulation of expression of immunoglobulin genes.
Production and characterization of T cell clones specific for mouse hepatitis virus, strain JHM: in vivo and in vitro analysis.
Authors: Woodward J G , Matsushima G , Frelinger J A , Stohlman S A
Published: December 2022

Mouse hepatitis virus, strain JHM, is a coronavirus capable of inducing both acute and chronic central nervous system disease. To characterize the cell-mediated immune response directed toward JHMV antigens, we have generated and analyzed the properties of four T cell clones reactive to JHMV antigens. All four clones are of the Ly-1+2- phenotype and show an antigen-specific, I-Ab-restricted proliferative response that is dependent on antigen-presenting cells (APC). The fine specificity of I-A restriction was revealed by their differential recognition of JHMV antigen in the context of mutant I-A molecules present on B6.C-H-2bm12 APC. Two clones were unable to recognize JHMV antigen presented on bm12 APC, whereas a third clone showed an equivalent response with both bm12 and wild type B6 APC. In the analysis of the viral specificity, three of the clones showed distinct cross-reactions with MHV-2 and very little if any cross-reaction with MHV-A59. In addition, three of three clones tested responded well to sucrose gradient-purified, UV-inactivated JHMV, indicating that their specificity is directed toward virus structural rather than nonstructural proteins. Finally, two of the clones were analyzed for their ability to induce a DTH reaction when injected together with antigen into the footpads of various mouse strains. Consistent with the in vitro data, both clones produced a marked DTH response only in those strains carrying the I-Ab genotype. These data demonstrate, both in vivo and in vitro, that at least one component of the cell-mediated immune response to JHMV is characterized by an Ly-1+2-, I-A-restricted T cell specific for JHMV structural proteins.
Two new monoclonal antibodies (LN-1, LN-2) reactive in B5 formalin-fixed, paraffin-embedded tissues with follicular center and mantle zone human B lymphocytes and derived tumors.
Authors: Epstein A L , Marder R J , Winter J N , Fox R I
Published: December 2022

Two monoclonal antibodies (LN-1, LN-2) reactive with B lymphocytes in B5 formalin-fixed, paraffin-embedded tissue sections have been produced by utilizing cell extracts from pokeweed mitogen-stimulated peripheral blood lymphocytes and diffuse histiocytic lymphoma SU-DHL-4 cells, respectively. Both monoclonal antibodies were initially identified by indirect immunofluorescence screening techniques on paraformaldehyde-acetone-fixed cell preparations. Specificity screens with 36 well-characterized human lymphoma and leukemia cell lines showed that both LN-1 and LN-2 stained cell lines of B cell lineage but were unreactive with those of T cell or, with one exception, myeloid derivation. Null cell acute lymphoblastic leukemia cell lines were found to be LN-2+ but LN-1-. The B cell specificity of these reagents was confirmed on 15 lymphoma and 17 leukemia biopsy specimens by using indirect immunofluorescence techniques. Immunoperoxidase staining of sections from B5-fixed, paraffin-embedded human lymphoid tissues showed that LN-1 bound to the cell membrane and cytoplasm of germinal center cells whereas LN-2 stained the nuclear membrane and cytoplasm of germinal center and mantle zone B lymphocytes as well as interfollicular histiocytes and thymic medullary dendritic cells. Both monoclonal antibodies failed to stain cortical thymocytes, lymph node T cells, and peripheral blood T and myeloid cells. Immunoperoxidase staining of 20 nonlymphoid human organs and tissues revealed that LN-1 reacted positively with red blood cell precursors of the bone marrow, ciliated epithelial cells of the bronchus, distal tubular cells of the kidney, and ductal cells from several organs including the breast and prostate. In contrast, LN-2 was unreactive with all human nonlymphoid organs and tissues including the bone marrow. Indirect immunofluorescence staining of a panel of 26 solid tumor cells lines showed that LN-1 was reactive with the majority of epithelium-derived cell lines, glioblastomas, and astrocytomas but was unreactive with neuroblastomas, small cell carcinoma of the lung, and sarcomas. LN-2 was unreactive with 25 of 26 of the solid tumor cell lines by these techniques. Immunobiochemical studies have shown that LN-1 recognizes a cell surface sialoantigen whereas LN-2 is directed against a 35,000 dalton nuclear membrane protein. Because of their high specificity for B cell tumors and their ability to stain B5-fixed, paraffin-embedded tissues, LN-1 and LN-2 are useful reagents for the diagnosis and classification of the human lymphomas and leukemias.
Description of a murine B lymphoma tumor-specific antigen.
Authors: Koch S , Zalcberg J R , McKenzie I F
Published: December 2022

By using monoclonal antibodies, a tumor-specific antigen (TSA 41.5) was detected on the cell surface of a B lymphoma CH-1 tumor variant, CH-1.1. This antigen is not expressed by normal lymphocytes (spleen cells, lymph node cells, thymocytes, bone marrow cells, or blast cells) of B10.A mice, the host strain of CH-1.1, or by the CH-1 lymphoma. Immunoprecipitation and biochemical characterization of TSA 41.5 with the use of two-dimensional gel electrophoresis showed this antigen to be a surface protein of CH-1.1 cells with an Mr of 80k and pI of 4.6. TSA 41.5 is not related to the murine transferrin receptor, and not to gp70, a viral envelope protein expressed by CH-1.1 cells, shown by comparative peptide map analysis of these three proteins. TSA 41.5 is a surface antigen unique to the CH-1.1 tumor, which is not expressed by the 19 different murine tumor lines that were tested nor by spleen cells of 15 independent mouse strains. In addition, treatment of spleen cells with bacterial lipopolysaccharide did not induce the expression of TSA 41.5. These characteristics of TSA 41.5 make it unlikely to be a product of viruses. Additional evidence against TSA 41.5 being a viral protein was obtained by the observation that antisera against viral proteins could not block the binding of the anti-TSA monoclonal antibody to its antigen. In vitro treatment of CH-1.1 cells with anti-TSA monoclonal antibody specifically inhibited the in vitro growth of the tumor cells in a dose-dependent fashion. The CH-1.1 tumor and monoclonal antibodies could be a useful murine model system for the exploration of the use of monoclonal antibodies for the in vivo treatment of cancer.
Modulation of polyclonally activated human peripheral B cells by aggregated IgG and by IgG-binding factors: differential effect on IgG subclass synthesis.
Authors: Bich-Thuy L T , Revillard J P
Published: December 2022

The regulation of IgG subclass production by polyclonally activated human B cells was investigated by using two systems previously shown to selectively suppress the generation of IgG-containing cells (CC) but not that of IgMCC or IgACC. The first one involved a brief exposure of peripheral blood mononuclear cells (PBMNC) to heat-aggregated human IgG (Agg-IgG) followed by repeated washings and culture with untreated autologous PBMNC. The second one was achieved by addition of human IgG-binding factor(s) (IgGBF) prepared by affinity chromatography from supernatants of unstimulated PBMNC. Pokeweed mitogen (PWM) and Nocardia opaca delipidated cell mitogen (NDCM) were used as polyclonal B cell activators. The latter can induce the terminal differentiation of peripheral B lymphocytes into plasma cells in the absence of helper T cells. After 6 days of culture, the number of cells containing IgM, IgG, or IgA was determined by direct immunofluorescence, and that of cells containing IgG1, IgG2, IgG3, or IgG4 was determined by indirect immunofluorescence with the use of subclass-specific monoclonal antibodies. After stimulation with PWM, exposure of PBMNC to Agg-IgG resulted in a selective diminution of the number of IgG4CC. With NDCM-stimulated cultures the same procedure induced a selective suppression of the generation of IgG2CC and IgG4CC. Conversely, the addition of IgGBF at the third day of culture was found to induce a 30 to 40% decrease in the number of cells containing each of the four IgG subclasses. Because of their differential pattern of IgG subclass suppression, Agg-IgG and IgGBF are likely to trigger distinct regulatory pathways.
Supplemental zinc restores antibody formation in cultures of aged spleen cells.
Authors: Winchurch R A , Thomas D J , Adler W H , Lindsay T J
Published: December 2022

The in vitro production of antibody to T-dependent antigens is depressed as a function of age. The addition of supplemental zinc to cultures of cells from young adult, middle-aged, and immunodepressed-aged mice enhances antibody production. The responses of young mice are enhanced 30 to 40%, but the responses in cultures from aged mice are increased 5- to 12-fold and the maximum response achieved is often equivalent to that of young, fully competent mice. The addition of zinc at varying times during the culture period suggests that zinc affects early events in the activation of antibody-forming cells.
B cell activation. V. Differentiation signaling of B cell membrane depolarization, increased I-A expression, G0 to G1 transition, and thymidine uptake by anti-IgM and anti-IgD antibodies.
Authors: Cambier J C , Monroe J G
Published: December 2022

Although studies have suggested that B cell membrane-associated IgM and IgD may function differently in the generation of humoral immune responses, this difference has proven difficult to define precisely. Toward a better understanding of the function of these antigen receptors, we compared the ability of F(ab')2 fragments of antibodies specific for mouse Fab, IgM, and IgD to induce early changes in B cell physiology that are indicative of cell activation. Specifically, we assessed the ability of membrane IgM (mIgM) or membrane IgD (mIgD) or both in combination to transduce signals that result in membrane depolarization, increased I-A expression, G0 to G1 transition, and thymidine uptake. Results indicate that at low levels of anti-receptor antibody (0.01 to 0.1 microgram/ml) both isotypes transduce signals for the induction of membrane depolarization and increased I-A expression by a significant proportion of B cells, i.e., Ig+ cells. Higher concentrations of ligand (5 to 25 micrograms/ml) are required to induce a significant proportion of cells to undergo G0 to G1 transition and increase thymidine uptake. At optimal concentrations, anti-IgM and anti-IgD induce membrane depolarization and increased I-A expression by 80 to 90% of B cells. Although mIgM also transduces signals sufficient for G0 to G1 transition by 80 to 90% of B cells, mIgD transduces signals sufficient for G0 to G1 by only about 40% of B cells. A similar difference exists for the induction of thymidine uptake. Thus, results suggest that membrane Ig may transduce two different signals depending upon the amount of ligand that they bind. Although in some B cells mIgD can transduce both signals resulting in membrane depolarization and progression into G1, in most cells mIgD transduces only the first signal, resulting in membrane depolarization and increased I-A expression.
Recognition and response to alloantigens in vivo. I. Negative and positive selection of MLR reactivity in murine peripheral blood lymphocytes to major histocompatibility complex and Mls antigens.
Authors: Ryan J J , Ahmed A , Kind P D , Thompson C B , Berning A K , Sell K W
Published: December 2022

Specific mixed lymphocyte reaction (MLR) responsiveness to allogeneic major histocompatibility complex (MHC), or minor lymphocyte-stimulating (Mls) determinants, was depleted in the peripheral blood lymphocytes (PBL) obtained from mice 24 to 48 hr after i.v. injection of 5 to 7.5 X 10(7) MHC or Mlsa-incompatible spleen cells, respectively. Results of cell mixture experiments suggest that the generation of suppressor cells was not the explanation for this specific reduction in MLR proliferation occurring with these PBL responder cells. To gain additional insight into parameters involved in the recognition of allodeterminants in vivo, experimental manipulations of the host environment and donor cell inoculum utilized in the negative selection procedure were employed. For example, removal of the spleen in the recipient animal, an anatomic site in which injected allogeneic cells and corresponding host antigen-reactive cells (ARC) are trapped, still permitted the specific depletion in murine PBL of host ARC for donor foreign MHC antigens. This finding may implicate other sites such as the liver where unprimed host alloreactive clones are trapped. In addition, irradiation of allogeneic donor cells significantly reduced their capacity to trap alloreactive T cell clones in vivo, whereas heat treatment of the donor cells completely eliminated this ability, even though the Ia determinants were still expressed, measured by flow cytometry. After the negative selection period, kinetic analysis of proliferation showed that 3, 4, or 5 days after injection of MHC-incompatible allogeneic spleen cells, the PBL of the recipient showed specific hyperresponsiveness to the MHC-haplotype of the donor cells. Interestingly, these primed PBL responder cells had the volume distribution of small resting cells; thoracic duct lymphocytes (TDL), positively selected by adoptive transfer of T cells to irradiated semiallogeneic recipients, are reported to be mainly blast cells. In contrast to the MLR hyperresponsiveness that results from priming with MHC-incompatible splenocytes, PBL, obtained at these later time points from mice primed with Mlsa-incompatible, H-2-compatible splenocytes, showed complete unresponsiveness in MLR to these Mlsa-bearing stimulator cells, as well as some nonspecific reduction in proliferation to MHC-incompatible stimulator cells regardless of their Mls genotype.(ABSTRACT TRUNCATED AT 400 WORDS)
Establishment of three PPD-reactive helper T cell clones with distinct functions in B cell activation.
Authors: Sano Y , Yamada G , Dobashi K , Mizuochi T , Hamaoka T , Takatsu K
Published: December 2022

Three PPD-reactive long-term cultured helper T cell clones were established from Mycobacterium tuberculosis (Tbc)-primed spleen cells. Clones B11.15 and B12.F were derived from C57BL/6 mice, and clone D-2 was originated from DBA/2Ha mice that have an X-linked recessive inheritance of T cell-replacing factor (TRF) unresponsiveness. Proliferative responses of these cloned T cells were induced by stimulation with PPD in a dose-dependent manner only when I-A-subregion compatible antigen-presenting cells (APC) were present. These three T cell clones have distinct helper functions in B cell activation. Clone B11.15 activated DNP-primed B cells to induce anti-DNP IgM plaque-forming cell (PFC) responses only when high amounts of PPD (5 micrograms) were added to the culture in a major histocompatibility complex (MHC)-unrestricted manner (factor-mediated interaction), whereas stimulation with a low amount of DNP-PPD (0.05 microgram) was ineffective. On the other hand, clone D-2 triggered B cells in the presence of a low amount of DNP-PPD in a MHC-restricted manner (cognate interaction). Significant helper activity of D-2, however, was not observed in the presence of high amounts of PPD. Clone B12.F was able to activate B cells in the presence of either DNP-PPD or PPD. Moreover, both B11.15 and B12.F produced helper factor(s) such as TRF by stimulation with high amounts of PPD in the presence of syngeneic APC, whereas D-2 did not produce measurable helper factor(s) under the same conditions. These results suggest that at least three distinctly functioning PPD-reactive helper T cells can be generated by active immunization with Tbc in vivo. T-B cell interaction between distinctly functioning T cell clones and B cells from (DBA/Ha X C57BL/6) (DB6)F1 male or female mice was then examined. B cells from DB6F1 female mice were triggered by both B11.15 and B12.F in a factor-mediated manner and were also activated with B12.F or D-2 in cognate manner. On the other hand, B cells from DB6F1 male mice, which are TRF low responders, were activated by B12.F or D-2 only through cognate interaction, and they failed to cooperate with B12.F or B11.15 in factor-mediated manner. These findings further suggest that B cells can be triggered by at least two distinct helper T cell subpopulations via respective pathways (cognate interaction and factor-mediated interaction).(ABSTRACT TRUNCATED AT 400 WORDS)
Antigen-reactive cloned helper T cells. II. Exposure of murine cloned helper T cells to IL 2-containing supernatant induces unresponsiveness to antigenic restimulation and inhibits lymphokine production after antigenic stimulation.
Authors: Wilde D B , Prystowsky M B , Ely J M , Vogel S N , Dialynas D P , Fitch F W
Published: December 2022

Cloned murine helper T lymphocytes (HTL) reactive to alloantigen or to ovalbumin (OVA) become unresponsive to antigenic restimulation after exposure to antigen or to culture supernatant fluids (SF) containing multiple lymphokine activities. Unresponsiveness is manifest by a failure of antigen-stimulated cells to incorporate thymidine or to produce lymphokines after antigenic challenge. Antigen-unresponsive HTL, however, will incorporate thymidine when exposed to an exogenous source of interleukin 2 (IL 2). The duration of unresponsiveness to antigen is correlated with the concentration of IL 2 in SF to which the cloned HTL had been exposed. Chromatographic fractionation of IL 2-containing supernatant from EL-4 thymoma cells (EL-4 SF) yielded a pool of SF that was enriched for IL 2 activity. Exposure of HTL to lymphokines contained in this pool induced unresponsiveness to antigen that was comparable to that observed when HTL were exposed to unfractionated EL-4 SF. Unresponsiveness to antigen also developed after cloned HTL were stimulated with concanavalin A (Con A) or with OVA and syngeneic splenic filler cells. We have used monoclonal antibody (mAb) GK1.5 (anti-L3T4) to investigate the role of lymphokine production in the induction of unresponsiveness. This antibody did not inhibit IL 2-induced thymidine incorporation by cloned HTL, and did not inhibit the induction of unresponsiveness after exposure of cloned HTL to EL-4 SF. In the presence of mAb GK1.5, however, HTL that were stimulated with Con A or OVA did not become unresponsive to antigenic restimulation, an effect that was overcome by the addition of EL-4 SF. These results suggest that HTL become unresponsive to antigen after exposure to IL 2-containing SF, and that stimulation by antigen or Con A can induce the unresponsive state by virtue of stimulating lymphokine production.
A shared antigenic specificity of human large granular lymphocytes and precursors of NK-like and allospecific cytotoxic effector cells.
Authors: Nieminen P , Saksela E
Published: December 2022

We describe here a new monoclonal IgG1 antibody, NK-9, which recognizes practically all large granular lymphocytes (LGL), the mediators of human NK activity. This antibody also reacts with an originally noncytotoxic, non-LGL lymphocyte population that contains precursors of the inducible NK-like and allospecific cytotoxicity in the blood. The NK-like effectors are also NK-9+. The results thus suggest a close precursor relationship of endogenous NK cells, NK-like cells, and allospecific CTL in the blood. NK-9 antibody does not crossreact with B cells, monocytes, or granulocytes or with any of a large panel of cultured cell lines tested, including T and B cell as well as null cell lymphoma-leukemia lines. It does not inhibit the functional properties of the positive cells. The antigens for NK-9 antibody on the cell surface have m.w. of 190, 200, and 220 kD, and at least the former two appear to be glycosylated. The non-LGL, noncytotoxic NK-9+ cells contain only the 190 kD antigen, but the other two appear concomitantly with the induced NK-like cytotoxic activity on the cells. The NK-9-detected antigens are retained after formaline fixation, and NK-9 antibody can thus be used to stain normal formaline-fixed, paraffin-embedded histologic sections.
Regulation of natural killer cell activity by macrophages in the rheumatoid joint and peripheral blood.
Authors: Combe B , Pope R , Darnell B , Kincaid W , Talal N
Published: December 2022

Recently, in another study, we observed that indomethacin, a prostaglandin synthetase inhibitor, significantly increased NK activity in both normal and rheumatoid arthritis (RA) peripheral blood (PB) but not in RA synovial fluid (SF). Because macrophages are a major source of prostaglandins, we examined the effect of macrophage-enriched adherent cells (AC) on NK activity as measured by a 3-hr Cr-release assay with K 562 cells. The removal of AC resulted in increased (p less than 0.01) NK activity in both normal and RA PB. In contrast, the removal of AC from RA SF resulted in a significant decrease (p less than 0.001) of NK activity. By using only nonadherent cells (NAC), NK activity in RA SF and synovial tissue (ST) was significantly reduced when compared to autologous RA PB (p less than 0.001). Enhancement of NK activity of SF NAC by both poly I:C and IL 2 was not dependent on AC. Mixing experiments demonstrated that the addition of synovial AC for 16 hr increased NK activity of synovial NAC to a level similar to that of unseparated mononuclear cells, whereas autologous PB AC suppressed NK activity of PB NAC. PB AC, when added to SF NAC, also increased NK activity. Supernatants from synovial mononuclear cells were stimulatory of synovial NAC NK activity, whereas normal PB mononuclear supernatants were suppressive. These observations document 1) a significant reduction of NAC-mediated NK activity in the rheumatoid joint as compared to PB from the same patient, and 2) that AC modulate NK activity differently in the rheumatoid joint as compared to RA or normal PB.
A unique series of lymphomas related to the Ly-1+ lineage of B lymphocyte differentiation.
Authors: Davidson W F , Fredrickson T N , Rudikoff E K , Coffman R L , Hartley J W , Morse H C
Published: December 2022

Six new B lineage lymphomas of NFS mice established in primary tissue culture were examined for a number of phenotypic, functional, virologic, and molecular genetic characteristics. Two of the tumors and their cloned derivatives bore surface markers characteristic of B cells, whereas four tumors resembled pre-B cells. One of the B cell and two of the pre-B cell lymphomas had rearrangements of both heavy and light chain immunoglobulin genes, confirming their designation as B-lineage lymphomas. All the tumors but one were Ly-1+, indicating that Ly-1 may be expressed by some pre-B cells as well as some B cells. In addition, one pre-B cell lymphoma was Mac-1+. MCF murine leukemia viruses obtained from two of the tumors did not accelerate development of B-lineage lymphomas in NFS mice.
Allotypes of murine factor B controlled by a locus within the S region of the H-2 complex.
Authors: Natsuume-Sakai S , Kaidoh T , Sudo K , Takahashi M
Published: December 2022

Bf protein was directly precipitated from a mixture of EDTA-plasma from 13 different, inbred strains and rabbit IgG anti-mouse Bf, and was isolated by SDS-PAGE. The gel pieces containing Bf protein were digested by trypsin after labeling with 125I-Na, and then peptides of Bf protein with each mouse strain were compared by two-dimensional peptide mapping. The results for the peptide patterns with the standard B10 congenic strains (all of which Bf phenotypes had been already designated as Bf.1 because of the identical isoelectric point values) revealed two distinct peptide patterns. The peptide pattern of Bf protein appeared to be identical in each of the inbred strains, but only one of the 36 spots in the fingerprint observed either with B10 or B10.BR (designated tentatively as Bf.1 (b.k.) ) was not detected in that from B10.D2 and B10.S (designated tentatively as Bf.1 (d.s.) ). The results of peptide mapping with intra-H-2 recombinant inbred strains show that this structural variant was mapped to the S region, which is direct evidence that allotypes of murine Bf are encoded by a structural gene within the MHC.
The blockade of Fc receptor-mediated clearance of immune complexes in vivo by a monoclonal antibody (2.4G2) directed against Fc receptors on murine leukocytes.
Authors: Kurlander R J , Ellison D M , Hall J
Published: December 2022

To evaluate the feasibility of using a monoclonal anti-Fc receptor antibody to alter Fc receptor function in vivo, the disappearance of radiolabeled human serum albumin-rabbit anti-human serum albumin (HSA-anti-HSA) complexes was studied in mice before and after the infusion of 2.4G2, a monoclonal antibody (developed by J. Unkeless). 2.4G2 specifically binds to Fc receptors on mouse macrophages. Under standardized conditions, 6.1% of an i.v. administered dose of anti-HSA was sequestered in the liver of B6/D2J mice. When HSA-anti-HSA complexes were administered, 53.4% were sequestered. If 8 micrograms/g body weight of 2.4G2 was infused i.p. 1.5 hr before HSA-anti-HSA, only 13.7% of the infused complexes were sequestered in the liver. The inhibition in Fc receptor-mediated sequestration produced by this dose of antibody persisted for at least 24 hr. A dose of 1 to 2 micrograms/g was sufficient to inhibit sequestration by 50%. Animals receiving daily injections of 2.4G2 cleared immune complexes from their blood much more slowly than untreated animals. Because 2.4G2 was not cytotoxic to peritoneal macrophages in vitro in the presence of serum, and because comparable inhibition of Fc receptor function was observed in vivo in C5-deficient mice, blockade of function does not depend upon complement-mediated lysis of macrophages. The maximal degree of inhibition of Fc receptor function obtained by using intact 2.4G2 was about twice that observed by using Fab fragments of 2.4G2 to block receptors. In addition to its effect on Fc receptor function, 2.4G2 also had a small but significant inhibitory effect upon the clearance of 125I-labeled heat-aggregated HSA by the mononuclear phagocyte system both in intact and C5-deficient mice. We conclude that 2.4G2 is a potent inhibitor of IgG Fc receptor-mediated immune clearance in vivo.
The behavior of the solubilized receptor for immunoglobulin E in polyethylene glycol-detergent solutions: characterization and potential applications.
Authors: Rivera J , Rivnay B , Furuichi K , Isersky C
Published: December 2022

We studied the effect of polyethylene glycol (PEG) on the solubility of the receptor for immunoglobulin E (IgE) in non-ionic detergent extracts of rat basophilic leukemia (RBL) cells. We found that the precipitation patterns of free and IgE-bound receptor were identical but differed from that of unbound IgE. Thus, 85 to 95% of the free receptor and the IgE-receptor complexes precipitated at 13% PEG in the presence of 0.5% Nonidet P-40, whereas 95% of the unbound IgE remained soluble. A similar degree of differentiation between the precipitation of receptor-bound and unbound IgE was found when we used extracts and PEG solutions prepared with several non-ionic and/or neutral detergents. The intact IgE-receptor complex with the full complement of subunits (alpha, beta, gamma) precipitated more efficiently than the IgE-alpha-chain-complex. The presence of phospholipids, which were previously shown to be important for preservation of the association between the receptor subunits, enhanced the efficiency of precipitation of the IgE-receptor complex. The presence of PEG also had an effect on the solubility of cellular phospholipids and some of the detergents, although the effect of PEG on either could not be directly related to its effect on the solubility of the IgE-receptor complex. The radioiodinated receptor for IgE, much like other radioiodinated RBL cell membrane proteins, was soluble (greater than or equal to 95%) at approximately 7% PEG but could be specifically and efficiently precipitated from crude cell extracts, in the presence of 7% PEG upon the addition of anti-receptor immunoglobulins alone. Using mouse anti-dinitrophenyl IgE antibody, we found that unlike unbound antigen (DNP-BGG) or the IgE-receptor complex, the detergent-solubilized DNP-BGG-IgE-receptor complex was insoluble at 7% PEG. Consequently, PEG can be employed in assays to quantitate the soluble receptor, and to immunoprecipitate it specifically and directly. Moreover, the use of PEG can facilitate the distinction between unbound antigen and antigen-IgE-receptor complex as well as between the latter and IgE-receptor complex.