bluuebunny/crossref_metadata_2025_split huggingface dataset:
Association of various T cell-surface molecules with the cytoskeleton. Effect of cross-linking and activation.
Authors: Geppert T D , Lipsky P E 
Published: December 2022

Abstract
The association of various surface molecules with the cytoskeleton in resting peripheral blood T cells was examined by assaying the capacity of detergent to solubilize them. Cytoskeletal association was assessed by staining T cells with a fluorescein-conjugated mAb, resuspending the cells in buffer with or without the nonionic detergent, NP-40, and determining the capacity of the detergent to remove the mAb from the cell surface by using flow microfluorimetry. MAb to CD3, the TCR, and CD45 were completely removed from the cell surface by detergent. In contrast, 7 to 50% of mAb to CD2, CD4, CD8, CD11a/CD18, CD44, and class I MHC molecules were resistant to detergent solubilization, demonstrating that a fraction of these molecules was constitutively associated with the cytoskeleton. The effect of cross-linking these molecules with a mAb and a secondary goat anti-mouse Ig was also examined. Cross-linking CD3 or the TCR induced cytoskeletal association of these molecules. In addition, cross-linking increased the fraction of CD2, CD4, CD8, CD11a/CD18, CD44, and class I MHC molecules that was associated with the cytoskeleton. In contrast, cross-linking CD45 did not induce an association with the cytoskeleton. The effect of T cell activation on the cytoskeletal association of these molecules was also examined. Stimulation of T cells with ionomycin and PMA greatly increased the expression of CD2 and CD44 without increasing the number of molecules associated with the cytoskeleton. Stimulation with PMA alone had no effect on the expression of CD2 or CD44, but was found to decrease the percentage of these molecules associated with the cytoskeleton. Stimulation with ionomycin and PMA increased both the expression of class I MHC molecules and the number of molecules associated with the cytoskeleton proportionally. Finally, stimulation with ionomycin and PMA decreased CD3 expression, but increased the number of CD3 molecules associated with the cytoskeleton. The data establish a pattern of cytoskeletal association of T cell-surface molecules that is a characteristic of each individual molecule and can be altered by cross-linking. Moreover, the results indicate that the association of various T cell surface molecules with the cytoskeleton is a dynamic process that varies with the state of activation and or differentiation of the cells.
Regulation of IgM and IgD expression in human B-lineage cells.
Authors: Kerr W G , Hendershot L M , Burrows P D 
Published: December 2022

Abstract
IgD is thought to function primarily as an Ag receptor that is expressed, together with IgM, only on mature B lymphocytes. This differentiation stage-specific expression of IgD has been well characterized in mice, where delta mRNA is detected only in mature IgM/IgD B cells. Humans, in contrast to mice, have significant levels of serum IgD, suggesting that the regulation of this isotype might differ between the two species. Therefore, we examined the regulation of both IgM and IgD expression in cell lines encompassing the spectrum of human B lineage development. Surprisingly, two species of delta mRNA could be found at all differentiation stages -from mu+ pre-B cell to IgM-secreting plasmablast. These mRNA are translated to yield the membrane and secretory forms of delta. The membrane delta-chain: secretory delta-chain ratio did not necessarily reflect the membrane mu-chain:secretory mu-chain ratio in the same cell line, implying that different mechanisms are involved in the selection of membrane vs secretory mu- and delta-chains. The delta-chains synthesized in pre-B cells were degraded, but in more mature cell types IgD could be stably expressed and secreted. Exceptions to this panlineage synthesis of delta-chains were, however, observed in two of the B cell lymphomas, where delta expression was prevented by transcriptional and posttranscriptional mechanisms. The presence of delta-chain in pre-B cells and the secretion of IgD by more mature cells suggest that IgD may have immunoregulatory roles throughout B cell differentiation. These studies also indicated that the bias toward secretory mu-chain production that occurs in human IgM secreting cells results from posttranscriptional regulation. In addition, we have identified a B cell line that synthesizes both normal-sized mu-chains and those with smaller apparent m.w. translation products of truncated mu mRNA.
Endocrine, neural, and immune control of secretory component output by lacrimal gland acinar cells.
Authors: Kelleher R S , Hann L E , Edwards J A , Sullivan D A 
Published: December 2022

Abstract
Androgens regulate the synthesis and secretion of secretory component (SC), the IgA antibody receptor, by acinar cells from the lacrimal gland. However, this hormone action may be susceptible to significant modification by other agents from the endocrine, nervous, or immune systems. To investigate the nature of this neuroimmunoendocrine interaction, the present study examined the impact of hormones, neurotransmitters, and lymphokines on basal and androgen-induced SC production by lacrimal gland acinar cells in vitro. Our results demonstrated that vasoactive intestinal peptide, the beta-adrenergic agonist, isoproterenol, PGE2, IL-1 alpha, IL-1 beta, and TNF-alpha significantly increased media SC levels in control or androgen-containing cell cultures. In contrast, the cholinergic agonist, carbachol, significantly decreased cellular SC output. These effects may be mediated through the agents' known capacity to alter intracellular cAMP levels. In support of this hypothesis, exposure of acinar cells to stimulators or analogues of cAMP resulted in a significant enhancement of SC production. Thus, these findings indicate that SC output in lacrimal tissue may be modulated by interactions between the endocrine, nervous and immune systems.
Amino acid residues in the T cell receptor CDR3 determine the antigenic reactivity patterns of insulin-reactive hybridomas.
Authors: Wither J , Pawling J , Phillips L , Delovitch T , Hozumi N 
Published: December 2022

Abstract
We examined TCR gene usage in a panel of beef insulin/I-Ad-restricted T cell hybrids obtained from BALB/c mice. These hybrids demonstrated several distinct patterns of reactivity defined by their ability to respond to species variants of insulin. Correlation of TCR-alpha and -beta-gene usage with these patterns of reactivity demonstrated that TCR gene usage was restricted within Ag reactivity groups. In particular, V-J junctional regions (CDR3 equivalent) were restricted with conserved junctional amino acid motifs present in both TCR-alpha- and -beta-chains. Comparison of TCR gene usage in hybrids expressing identical V alpha and V beta gene segments but demonstrating different patterns of reactivity revealed that changes in either J alpha and/or J beta gene segment usage could alter antigenic reactivity. Indeed, single or limited amino acid differences within the CDR3 region were sufficient to markedly alter fine specificity. These data demonstrate the critical role for CDR3 in determining antigenic reactivity in beef insulin-reactive hybrids and are compatible with the current model of TCR/peptide/MHC interaction.
Antiviral activity and dose optimum of recombinant macrophage colony-stimulating factor on herpes simplex genitalis in guinea pigs.
Authors: Ho R J , Chong K T , Merigan T C 
Published: December 2022

Abstract
The antiviral activity of recombinant human macrophage CSF (M-CSF) against genital herpes simplex virus type-2 (HSV-2) infection in guinea pigs was investigated. M-CSF stimulates proliferation of human and guinea pig peripheral blood monocytes, specifically the plastic adherent esterase-positive mononuclear cells. When anti-HSV-2 activity of M-CSF was evaluated in guinea pigs by 6 daily injection (s.c.) of M-CSF at various doses (5 x 10(5) to 7 x 10(7) U/kg), we found 2 x 10(6) U/kg to be the optimum dose for protective efficacy against primary HSV-2 infection. Either at a lethal, 5 x 10(5) pfu, or sublethal 5 x 10(4) pfu of virus challenge, animals treated with the optimum regimen of M-CSF exhibited lower herpetic lesion scores (p less than 0.005), and lower mortality (p less than 0.025) than animals in placebo group. M-CSF treatment increased the HSV-infected cell killing activities of plastic-adherent mononuclear cells, indicating that in vivo administration of M-CSF may activate the antiviral effects of guinea pig macrophages that may play a role in protection against severity and mortality of herpetic disease.
Murine IgG isotype responses to the Plasmodium cynomolgi circumsporozoite peptide (NAGG)5. I. Effects of carrier, copolymer adjuvants, and lipopolysaccharide on isotype selection.
Authors: Kalish M L , Check I J , Hunter R L 
Published: December 2022

Abstract
The tandem repeat peptide of the circumsporozoite protein of Plasmodium cynomolgi, (NAGG)5, conjugated to BSA or Salmonella flagella, was injected into mice with block copolymer and other adjuvants. The flagella carrier preferentially stimulated IgG2a antibodies to (NAGG)5 that constituted as much as 85% of the total IgG antibody whereas the BSA carrier stimulated as much as 98% IgG1. The distribution of isotypes of antibody to (NAGG)5 was also modified by using the copolymer adjuvants L121 or L141, either alone, or especially in combination with a nontoxic LPS. L121 or L141 increased the proportion of IgG2a and IgG2b antibodies to (NAGG)5 after immunization with (NAGG)5-BSA whereas LPS stimulated further increases in IgG2 antibodies (up to 69% of the total IgG). The hapten density, physical form of emulsion, and route of immunization further affected the isotypes produced in this study.
Role of listeriolysin-O (LLO) in the T lymphocyte response to infection with Listeria monocytogenes. Identification of T cell epitopes of LLO.
Authors: Safley S A , Cluff C W , Marshall N E , Ziegler H K 
Published: December 2022

Abstract
Using a murine model, we investigated the role of the bacterial exotoxin listeriolysin O (LLO) in cellular immunity to Listeria monocytogenes. A correlation between LLO production by infecting bacteria and generation of protective immunity to virulent LLO-producing bacteria was noted. Using isogeneic hemolysin (Hly+ or Hly-) strains of L. monocytogenes, we demonstrated that LLO production by infecting bacteria is required to elicit T cells reactive both to bacteria-associated Ag and to the secreted LLO molecule as measured by IL-2 production in vitro. Distinct sets of T cells specific for largely nonoverlapping pools of antigenic determinants represented by LLO and cell-associated Ag (heat-killed L. monocytogenes) are generated after infection. We have used models for prediction of T cell epitopes based on primary structure of LLO, and synthetic amphipathic LLO peptides were evaluated as Ag in vitro or as immunogenes in vivo. Infection of several strains of mice (H-2k and H-2d) with LLO-producing L. monocytogenes resulted in the generation of T cells that could respond consistently to two peptides, LLO 215-234 and LLO 354-371. Mouse strains lacking expression of I-E molecules (e.g., B10.A(4R) and C57BL/6) responded to LLO but not to the peptides tested. With C3HeB/FeJ mice, antibodies to I-Ek blocked the presentation of LLO 215-234. The importance of the N-terminal portion of LLO 215-234 was evidenced by the drastic reduction in antigenic activity of truncated peptides (e.g., LLO 221-234 and LLO 224-234). LLO 215-234, the strongest and most consistent activator of T cells from L. monocytogenes-immune mice, fit well some models for antigenic peptides in several ways. It could be predicted to form an amphipathic alpha-helix, it contained multiple "Rothbard motifs" (charged residue or glycine, two or three hydrophobic amino acids and then a glycine or polar residue), it had a net charge of +2, and it contained the correct spacing of amino acids (five to six residues between a hydrophobic and basic amino acid) that is characteristic of I-Ek-binding peptides. Immunization with 8 of 10 synthetic LLO peptides generated T cells that recognized the immunizing peptide in vitro, but such T cells were only poorly reactive with LLO. Our results indicate that LLO is an important target Ag for stimulation of CD4+ L. monocytogenes-specific T cells, and that LLO 215-234 is antigenically dominant in C3HeB/FeJ mice.
Polymorphism of the human Ig VH4 gene family.
Authors: van Dijk K W , Sasso E H , Milner E C 
Published: December 2022

Abstract
In this study, we analyze the human VH4 gene family and find it to exhibit a level of polymorphism similar to that of the much larger VH3 family. A cloned VH4 probe detected an average of 10 hybridizing BgIII restriction fragments in genomic DNA derived from 75 unrelated individuals and a total of 15 distinct bands. Of these 15 restriction fragments, 12 were polymorphic, as demonstrated by band absence in some individuals. Oligonucleotide probes specific to CDR1 and CDR2 sequences of known VH4 genes detected limited numbers of bands and revealed sequence polymorphisms that correlated with several of the RFLP detected by the cloned probe. The prevalence of the individual polymorphic restriction fragments was highly variable, ranging from 1% to 97%, with a mean prevalence of 51%. These values resemble those previously observed among VH3 elements. Analysis of linkage disequilibrium suggests that most VH4 gene segments are in genetic equilibrium. These results indicate that the VH4 loci, like those of VH3, are dominated by relatively few, perhaps two to four, alleles/locus and further suggest that the haplotype organization of the human VH locus is very complex.
Specific release of granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-alpha, and IFN-gamma by human tumor-infiltrating lymphocytes after autologous tumor stimulation.
Authors: Schwartzentruber D J , Topalian S L , Mancini M , Rosenberg S A 
Published: December 2022

Abstract
Tumor-infiltrating lymphocytes (TIL) have been cultured from a variety of human tumors, and some melanoma TIL have demonstrated specific, MHC-restricted recognition of autologous tumor in short term lysis assays. The current study investigates cytokine release by TIL as an indicator of specific tumor recognition. We have identified two of four melanoma and one of seven breast carcinoma TIL cultures that specifically release granulocyte-macrophage-CSF, TNF-alpha, and IFN-gamma after autologous tumor stimulation. The other cultures either do not secrete cytokine or secrete cytokine in a nonspecific fashion. The amount of specific cytokine released is directly related to the number of TIL and stimulating tumor cells. Studies of TIL, from two melanoma patients, separated into CD4+ and CD8+ populations revealed that CD8+ cells were responsible for virtually all of the specific cytokine secretion, although both populations released cytokines when activated by immobilized anti-CD3 antibody. Specific cytokine release by CD8+ TIL was inhibited by anti-MHC class I mAb. Specific cytokine release was also detected from a CD4+ breast cancer TIL culture, and this was inhibited by anti-MHC class II mAb. The clinical significance of this specific mode of immune antitumor reactivity is currently under investigation.
Universal vaccine carrier. Liposomes that provide T-dependent help to weak antigens.
Authors: Garcon N M , Six H R 
Published: December 2022

Abstract
The design of a thymus-dependent synthetic vaccine that will provide a universal T cell epitope for B cell epitopes is described in this study. Simultaneous incorporation into liposomes of both a peptide recognized by Th lymphocytes and a lipophilic hapten and the IgG antibody responses to this hapten were assessed in outbred mice. DNP-aminocaproyl phosphatidylethanolamine (DNP-CapPE) is a well characterized T-independent hapten Ag. HA2 peptide derived from the hemagglutinin protein of influenza virus contains amino acid sequences recognized by Th and T cytotoxic lymphocytes. In addition, HA2 contains a sequence of hydrophobic amino acids near the carboxyl terminus, allowing its incorporation into liposomes. Results of immunization show that (i), when DNP-CapPE is carried by liposomes without the HA2 peptide, an IgM antibody response is induced, (ii) liposomes carrying both HA2 and DNP-CapPE elicit an IgG antibody response to DNP in a dose-dependent fashion for both HA2 and DNP, (iii) the liposomes must be processed intracellularly in order to elicit a response, (iv) the system leads to a memory response for DNP, and (v) all of the IgG subclasses are elicited. These data suggest that liposomes containing the HA2 peptide exhibit a T-dependent carrier effect for a T-independent Ag. The significance of these findings is discussed in conjunction with the characteristics of the liposome model used.
CD2 antigen mediated activation of the guanine nucleotide binding proteins p21ras in human T lymphocytes.
Authors: Graves J D , Downward J , Rayter S , Warne P , Tutt A L , Glennie M , Cantrell D A 
Published: December 2022

Abstract
T cell stimulation via the TCR complex (TCR/CD3 complex) results in activation of the guanine nucleotide binding proteins encoded by the ras protooncogenes (p21ras). In the present study we show that the activation state of p21ras in T lymphocytes can also be controlled by triggering of the CD2 Ag. The activation state of p21ras is controlled by GTP levels on p21ras. In T cells stimulation of protein kinase C is able to induce an accumulation of "active" p21ras-GTP complexes due to an inhibitory effect of protein kinase C stimulation on the intrinsic GTPase activity of p21ras. The regulatory effect of protein kinase C on p21ras GTPase activity appears to be mediated via regulation of GAP, the GTPase activating protein of p21ras. In the present report, we demonstrate that the TCR/CD3 complex and the CD2 Ag control the accumulation of p21ras-GTP complexes via a regulatory effect on p21ras GTPase activity. The TCR/CD3 complex and CD2 Ag are also able to control the cellular activity of GAP. These data demonstrate that p21ras is part of the signal transduction responses controlled by the CD2 Ag, and reveal that the TCR/CD3 complex and CD2 Ag control the activation state of p21ras via a similar mechanism.
Monoclonal antibody 4H12 recognizes subsets of adherent-lymphokine activated killer cells and splenic natural killer cells from pregnant and neonatal mice.
Authors: Linnemeyer P A , Pollack S B 
Published: December 2022

Abstract
Using a new mAb, 4H12, that recognizes a plasma membrane-associated Ag on granulated metrial gland cells, we identified subsets of murine NK cells in spleen cell-derived adherent lymphokine activated killer cells and in the spleens of neonatal and pregnant mice. In spleen cell adherent-lymphokine-activated killer cultures, 4H12 Ag was detected on a small subset of cells after 7 days culture and expression increased with time to 70% of the cells after 21 days culture. 4H12+ cells were large (up to 70 microns) and granular. The Ag was also detected in the cytoplasmic granules of some, but not all 4H12 surface positive cells. Coexpression studies indicated 4H12+ cells were predominantly positive for the NK1.1 and ASGM1 Ag, negative for the MAC-1 and F4/80 Ag, and +/- for the LGL-1 and CD3 Ag. Subsets of 4H12+/-, LGL-1+/- exhibited morphologic characteristics restricted to specific phenotypic subsets. The 4H12-/LGL-1+ subset was shown to contain the smallest, least granular cells, whereas the 4H12+/LGL-1+/- subsets contained the largest and most granular cells. Although 4H12 expression was negligible in the spleens of normal adult mice, spleen cells of neonatal and pregnant mice contained subsets of NK1.1+ cells that coexpressed 4H12. The 4H12+/NK1.1+ and 4H12-/NK1.1+ subsets displayed differential levels of NK1.1 expression. 4H12+ NK cells were NK1.1 low to high, whereas 4H12- NK cells were NK1.1 high only. The functional significance of subsets of NK cells in IL-2 culture and in the spleens of neonatal and pregnant mice remains to be elucidated.
CD2 can mediate TCR/CD3-independent T cell activation.
Authors: Ohno H , Ushiyama C , Taniguchi M , Germain R N , Saito T 
Published: December 2022

Abstract
T lymphocytes can be activated clonotypically through TCR/CD3 complex or polyclonally via the CD2 molecule. Whether CD2-mediated activation is dependent on TCR/CD3 expression or signaling is controversial. We have re-explored this issue by using a series of CD2-transfected, TCR/CD3 surface membrane-negative human and mouse T cells. Our results clearly show that such T cells can be triggered for IL-2 secretion and increases in intracellular Ca2+ through the CD2 molecule in the absence of surface expression of TCR/CD3 complexes. These responses are only observed when cells express high levels of CD2 and there is a critical threshold of CD2 expression necessary for such activation in the absence of CD3. Concomitant expression of TCR/CD3 complex markedly lowers the level of CD2 required for activation via the latter pathway. These results provide a clear resolution of the controversy concerning the requirement for surface CD3 expression in T cell activation through CD2 and further suggest a possible role for CD2 in activation of TCR/CD3-negative cells.
IL-4 increases IL-2 production by T cells in response to accessory cell-independent stimuli.
Authors: Tanaka T , Ben-Sasson S Z , Paul W E 
Published: December 2022

Abstract
Freshly prepared, highly purified T cells from naive mice failed to produce IL-2 in response to soluble anti-CD3 antibody or to Con A and produced only small amounts of IL-2 in response to anti-CD3 coated on the surface of microwells. IL-2 production in response to soluble anti-CD3 or to Con A required the addition of accessory cells. By contrast, the addition of IL-4 strikingly enhanced the production of IL-2 by plate-bound anti-CD3-stimulated T cells in the absence or the presence of added accessory cells. Furthermore, anti-IL-4 mAb inhibited IL-2 production by anti-CD3-stimulated T cells, which indicates that endogenously produced IL-4 was important in IL-2 production by T cells to plate-bound anti-CD3. The capacity of IL-4 to enhance and of anti-IL-4 to inhibit IL-2 production in response to plate-bound anti-CD3 was also observed with both unstimulated T cells and with T cells that had been previously stimulated with anti-CD3 antibody. When activated T cells were restimulated with anti-CD3, the effect of IL-4 in enhancing IL-2 production was detectable within 6 to 8 h after restimulation. The effect of IL-4 on IL-2 production was not due to prolongation of survival or to enhanced proliferation of T cells. Northern blot analysis showed that T cells treated with anti-CD3 plus IL-4 had more than 10-fold more IL-2 mRNA than did T cells treated with anti-CD3 plus anti-IL-4; this was observed within 6 h of stimulation under certain circumstances. The increased level of IL-2 mRNA by IL-4 was achieved without any change in message half-life, suggesting that IL-4 enhances transcriptional activation of the IL-2 gene in such cells. These results lead to the conclusion that IL-4 has a critical role in IL-2 production in response to accessory cell-independent stimuli (plate-bound anti-CD3 antibody), although it is not essential to IL-2 production in response to accessory cell-dependent stimuli (soluble anti-CD3 and Con A).