bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of macrophage C3b-mediated ingestion by syphilitic hamster T cell-enriched fractions.
Authors: Tabor D R , Azadegan A A , Schell R F , Lefrock J L
Published: December 2022

Macrophages are important for host defense against syphilitic infection. Our results show that C3b-mediated ingestion (C3bMI), a characteristic of activated macrophages, was inhibited in vitro by nonadherent cells from hamsters infected with Treponema pallidum subspecies endemicum. When macrophage target cells from normal, syphilitic, or lipopolysaccharide-treated animals were co-cultured with nonadherent cells derived from normal or syphilitic hamsters, noticeable differences were detected. Nonadherent syphilitic cells significantly suppressed macrophage C3bMI, whereas normal nonadherent cells displayed little or no suppressive activity. In general, macrophage C3bMI was reduced by nonadherent cells obtained throughout the course of syphilitic infection, although it eventually began to recover. The syphilitic nonadherent cells with maximum suppressive effect were those obtained from hamsters infected for 3 to 6 wk. The addition of treponemal antigens additionally inhibited C3bMI. The inhibitory effect of syphilitic nonadherent cells on either lipopolysaccharide-treated or syphilitic macrophages was sustained even when the nonadherent cells were removed from the cultures, and the effect continued for at least 72 hr thereafter. Fractionation of syphilitic nonadherent cell populations by two independent methods produced T cell-enriched preparations with significantly more suppressive activity than non-T cell-enriched preparations. These observations may account for the chronicity of syphilitic infection.
Human cytotoxic T cell clones directed against herpes simplex virus-infected cells. II. Bifunctional clones with cytotoxic and virus-induced proliferative activities exhibit herpes simplex virus type 1 and 2 specific or type common reactivities.
Authors: Yasukawa M , Zarling J M
Published: December 2022

Human cytotoxic T lymphocyte (CTL) clones directed against herpes simplex virus (HSV)-infected cells were generated after stimulation of peripheral blood lymphocytes (PBL) with HSV type 1 (HSV-1) and HSV type 2 (HSV-2). These CTL clones were studied with regard to HSV type specificity and with regard to whether they also express helper cell activity. Some clones, generated after stimulation with HSV-1, were cytotoxic for autologous cells infected with either HSV-1 or HSV-2 ("HSV type common clones"), whereas other clones lysed HSV-1-infected cells only ("type-specific clones"). Similarly, after HSV-2 stimulation, both HSV-2 specific and HSV type common clones were obtained, indicating the heterogeneity of human cytotoxic T cells to HSV. All CTL clones tested were found to be bifunctional in that they also proliferated in response to stimulation with HSV. The HSV type specificity of the proliferative response was identical to that of the cytotoxic activity of the clones. An HSV type common clone, when stimulated with either HSV-1 or HSV-2, and an HSV-1 specific clone, when stimulated with HSV-1 but not with HSV-2, produced a factor, presumably interleukin 2 (IL 2), which induced proliferation of CTLL, an IL 2-dependent T cell line, providing evidence that our HSV-directed CTL clones also express helper cell activity. CTL clones that we previously reported were restricted in cytotoxic activity by HLA class II DR-1 or MB-1 antigens were found, in this study, to be restricted in proliferative response to HSV by these same HLA antigens. These results suggest that our bifunctional T cell clones directed against HSV may recognize the same viral antigenic determinants and the same HLA antigens for both cytotoxic and virus-induced proliferative activities. This is the first demonstration of human HSV type specific and HSV type common T cell clones and HSV specific T cell clones with both cytotoxic and helper cell activities.
T cell responses to cleaved rabies virus glycoprotein and to synthetic peptides.
Authors: Macfarlan R I , Dietzschold B , Wiktor T J , Kiel M , Houghten R , Lerner R A , Sutcliffe J G , Koprowski H
Published: December 2022

The antigenic structure of the rabies virus glycoprotein has been studied. A limited number of fragments were obtained by cyanogen bromide (CNBr) cleavage of viral glycoprotein, and eight large peptides were isolated by using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. These were tested for their capacity to stimulate the proliferation of nylon wool-purified T cells obtained from spleens of rabies-immune A/J mice. Three peptides (Cr1, Cr2 plus Cr2A, and Cr3) stimulated antigen-specific proliferation, indicating that at least three T cell determinants of the native molecule are sequential or continuous in nature. Stimulation was also obtained with 27-residue and 13-residue synthetic peptides (designated R21 and R20, respectively) that included sequences towards the carboxy terminal end of Cr1, but not with synthetic peptides that included sequences of Cr2 and Cr3 (which are both glycosylated in virus-derived material). The intact viral glycoprotein and synthetic peptide R21 stimulated T lymphocytes with surface characteristics of helper cells, and induced the production of interleukin 2 by these lymphocytes. Synthetic peptides R20 and R21 also stimulated a minor population of Lyt-2-positive cells, which were not yet identified as either suppressor or cytotoxic T lymphocytes.
Sex and H-2 haplotype control the resistance of CBA-BALB hybrids to the induction of T cell lymphoma by Moloney leukemia virus.
Authors: Storch T G , Chused T M
Published: December 2022

CBA/N and CBA/CaHN have a significantly longer latent period than other inbred mouse strains between infection with Moloney murine leukemia virus and the appearance of T cell lymphoma. The genetic characteristics of this resistance have been analyzed in the F1 hybrids of CBA/N and CBA/CaHN with BALB H-2 congenic strains. Sexual phenotype and H-2 haplotype significantly influenced survival in the F1 hybrids of CBA/CaHN with BALB. In the F1 with BALB/cJ and BALB/cAnN (both H-2d), the males survived significantly longer than the females; but in the F1 with BALB.K (H-2k) and BALB.B (H-2b), the survival of males and females was the same. Survival was not prolonged by the recessive X-linked immunodeficiency gene xid or other genes on the CBA/N X-chromosome, because the (CBA/N X BALB/c)F1 male and the reciprocal (BALB/c X CBA/N)F1 male, which does not carry the CBA X-chromosome, were equally resistant. H-2 haplotype did not influence survival among the BALB H-2 congenics, and sex had little effect on the resistance of the CBA and BALB parents. These results demonstrate that a sex-dependent gene linked to H-2 significantly influences the expression of CBA genes for lymphoma resistance in the F1 hybrid with BALB.
Effects of ultraviolet radiation on human epidermal cell alloantigen presentation: initial depression of Langerhans cell-dependent function is followed by the appearance of T6- Dr+ cells that enhance epidermal alloantigen presentation.
Authors: Cooper K D , Fox P , Neises G , Katz S I
Published: December 2022

The effects of ultraviolet radiation (UV) on the immune parameters of human epidermis were studied. We determined the effects of both in vitro and in vivo UV on human epidermal cell surface markers and on epidermal immune function in the allogeneic epidermal cell-lymphocyte reaction (ELR). Epidermal cells obtained immediately after in vitro and in vivo UV exposure exhibited a dose-dependent decrease in alloantigen-presenting function in the ELR. This was not the result of a decrease in the number of T6+ Dr+ Langerhans cells but was due to their being less efficient at alloantigen presentation than equivalent numbers of Langerhans cells from unirradiated skin. The reduced stimulation in the ELR immediately after UV was not reversible by the addition of exogenous IL 1 or indomethacin and thus appeared to be due to a direct effect of UV on the alloantigen-presenting function of Langerhans cells. In contrast to this suppression of the epidermal immune function when epidermal cells were obtained immediately after UV, epidermal cells harvested 24 hr or later after in vivo UV exhibited a dose-dependent enhancement of allostimulatory capacity in the ELR that peaked 3 days after UV. The time course of the enhancement of allostimulation in the ELR after in vivo UV coincided with a decrease in the percentage of Langerhans cells and the appearance within the epidermis of T6- Dr+ cells, which are derived from the bone marrow, as evidenced by their expression of the bone marrow derivation markers HLe 1 and T200. Removal of Dr+ cells but not of T6+ cells from epidermal cell suspensions harvested 3 days after in vivo UV abrogated allostimulation in the ELR, demonstrating that the T6- Dr+ cells were responsible for the observed UV-induced enhancement of alloantigen presentation. Taken together, the results indicate that the timing and dosage of UV exposure are critical factors determining whether suppression or enhancement of epidermal immune function follows UV.
Defective IFN-gamma production in the human neonate. II. Role of increased sensitivity to the suppressive effects of prostaglandin E.
Authors: Wakasugi N , Virelizier J L , Arenzana-Seisdedos F , Rothhut B , Huerta J M , Russo-Marie F , Fiers W
Published: December 2022

Analysis of endogenous production and effects of exogenous addition of interleukin 2 (IL 2), leukotrienes (LT), and prostaglandin E (PGE) has been used to investigate the dysregulation responsible for impaired PHA-induced IFN-gamma secretion by cord blood leukocytes (CBL). The addition of LT or IL 2 could not reverse the IFN defect of CBL. The production of these two mediators was found to be normal in CBL cultures. CBL and control leukocytes from adult donors produced comparable amounts of PGE2. In contrast, sensitivity to the suppressive effects of PGE2 on IFN-gamma secretion was much higher with CBL than with control leukocytes. Treatment with indomethacin reversed the IFN-gamma defect with most CBL tested, and the addition of physiologic amounts of PGE2 to indomethacin-treated cultures resulted in a profound impairment of IFN-gamma production similar to that of untreated CBL cultures. Preincubation of CBL for 24 hr before PHA stimulation resulted in restoration of a normal sensitivity to exogenous PGE2, in parallel with correction of the IFN-gamma defect. Our observations suggest that the impairment of IFN-gamma secretion in neonates is not due to deficient amplification circuits, but is the consequence of an exaggerated cellular sensitivity to the suppressive effects of PGE produced endogenously in normal amounts.
Potential Z-DNA-forming elements in serum DNA from human systemic lupus erythematosus.
Authors: Van Helden P D
Published: December 2022

DNA fragments were isolated from serum of a patient with systemic lupus erythematosus. The majority of the DNA was between 150 and 250 base pairs in length. The DNA was cloned into phage M13, and 10 recombinants were sequenced. The average GC content of the DNA was higher than total human DNA (43% against 38%), with some fragments as high as 63%. This DNA is rich in alternating purine-pyrimidine segments that are potentially Z-DNA-forming regions.
Characterization of T lymphocyte subpopulations responsible for deficient interleukin 2 activity in patients with systemic lupus erythematosus.
Authors: Murakawa Y , Takada S , Ueda Y , Suzuki N , Hoshino T , Sakane T
Published: December 2022

Normal immunoregulation depends on a complex set of cellular interactions in which interleukin 2 (IL 2) appears to play an important role. We have examined the IL 2 activity in patients with systemic lupus erythematosus (SLE). IL 2 production by phytohemagglutinin (PHA)-stimulated T cells for 48 hr was measured by the ability of their culture fluid to induce proliferation of normal human T cells that had been activated for more than 20 days by PHA plus IL 2. To measure IL 2 responsiveness, T cells were blasted by preincubation with concanavalin A for 96 hr and stimulated for another 72 hr with lectin-free standard IL 2. SLE T cells failed to produce normal levels of IL 2 in vitro compared with normal control T cells. This failure resided in both OKT4+ and OKT8+ cells. Furthermore, the abnormality was due neither to soluble inhibitory factors produced by SLE T cells nor to active suppressor cells that might be induced by PHA-stimulation. Responsiveness to IL 2 of T cells from some, but not all, SLE patients was decreased significantly from that of normal controls. Absorption studies as well as studies with anti-Tac antibody demonstrated that the impaired responsiveness of T cells in the specific patients with SLE was due to inadequate expression of IL 2 receptors on the T cells upon activation. This defect was exclusively ascribed to the dysfunction of OKT4+, but not OKT8+, cells. The above defects in production of and responsiveness to IL 2 observed in patients with SLE were present at all times regardless of the disease activity or of corticosteroid therapy. Thus, the deficient IL 2 activity may be intrinsic to SLE lymphocytes and may contribute to impaired immunoregulation and to the development of SLE.
Subsets of Lyt-2+ cells defined by differential expression of the 9F3 antigen: alterations in mice of the lpr/lpr genotype.
Authors: Dumont F J , Habbersett R C , Coker L Z
Published: December 2022

Recently, we found that a novel murine cell surface antigen recognized by the 9F3 monoclonal antibody demonstrates T cell heterogeneity in normal mice and in congenic strains homozygous for the lymphoproliferation- and autoimmunity-inducing lpr gene. The objective of the present work was to further characterize this heterogeneity by studying the distribution of the 9F3 marker among Lyt-2+ T cells. By using dual parameter flow cytofluorometry analysis, at least two subsets of peripheral Lyt-2+ cells differing in 9F3 expression could be distinguished. In MRL/Mp-++, C57BL/6, and C3H/HeJ mice, 9F3- and 9F3+ cells accounted, respectively, for 65 to 80% and 20 to 35% of the whole Lyt-2+ population. Interestingly, in lpr/lpr-bearing congenic strains, a three- to fivefold selective increase in the frequency and absolute number of 9F3+ Lyt-2+ cells was found to take place during aging. Functional analysis of Lyt-2+ cells isolated by panning revealed that in +/+ mice, these cells respond better to phytohemagglutinin and concanavalin A than in lpr/lpr mice, indicating that phenotypic changes of Lyt-2+ cells correlate with functional changes. Further evidence for the functional relevance of 9F3-defined Lyt-2+ cell heterogeneity was provided by the study of sorted cells from +/+ mice, showing that 9F3- cells exhibit higher mitogenic reactivities than 9F3+ cells. This finding suggests that 9F3+ Lyt-2+ cells in +/+ and lpr/lpr mice are functionally homologous. Together, these data indicate that, in addition to the known expansion of a population of abnormal Lyt-2-T cells, mice of the lpr/lpr genotype have an alteration of their Lyt-2+ cell population, characterized by an imbalance of 9F3-defined subsets and decreased mitogenic reactivities.
Inhibition of human cytotoxic T lymphocyte activity in vitro by autologous peripheral blood granulocytes.
Authors: Petrie H T , Klassen L W , Kay H D
Published: December 2022

Peripheral blood granulocytes from normal healthy donors were found to reproducibly inhibit the cytolytic effector function of specifically sensitized cytotoxic T lymphocytes in vitro when co-incubated with these effector cells and target cells in 8 hr 51Cr release assays. Inhibition required intact granulocytes, was proportional to the number of granulocytes present, and was independent of granulocyte adherence, phagocytic function, and viability. Equivalent numbers of enriched normal or leukemic peripheral T lymphocytes did not cause inhibition of 51Cr release, and preincubation of granulocytes with effectors did not significantly alter viability or cytotoxic function. Because granulocytes can inhibit natural killer cell function in vitro, these data indicate that granulocytes can regulate diverse antigen-specific and spontaneous cytotoxic functions in vitro, suggesting that circulating granulocytes may have the potential for in vivo regulation of these cytotoxic effectors.
Human alloreactive CTL clones: loss and reacquisition of specific cytolytic activity can be regulated by "recombinant" interleukin 2.
Authors: Wee S L , Ochoa A C , Bach F H
Published: December 2022

In this communication, we demonstrate that human alloreactive CTL at the clonal level, can undergo reversible processes vis-a-vis lytic function: from a highly lytic phase to a nonlytic quiescent phase and back to a lytic phase. "Reversion" and reacquisition of function can be controlled by removing or adding back a chromatographically purified product of a human interleukin 2 gene (rIL 2) that appears to serve, either directly or indirectly, as a lysis-inducing signal for these CTL.
T cell-derived B cell growth factor(s) can induce stimulation of both resting and activated B cells.
Authors: Thompson C B , Schaefer M E , Finkelman F D , Scher I , Farrar J , Mond J J
Published: December 2022

The effects of a preparation containing partially purified, EL4-derived B cell growth factor(s) (BCGF) on B cell growth and proliferation have been examined by using B lymphocyte subpopulations separated on the basis of size. BCGF was found to maintain and enhance proliferation of a significant proportion of large activated B cells. In contrast, small resting B cells required the presence of BCGF and a second stimulus such as anti-IgM antibody (anti-mu) to be induced to proliferate. This disparity was not due to a lack of an effect of BCGF on small resting B cells. A factor contained within the partially purified EL4 supernatant produced time-dependent increases in cell size and RNA content in all subpopulations. These effects were independent of possible effects due to contaminating lymphokines such as interleukin 2 (IL 2), concanavalin A (Con A), and phorbol myristate acetate (PMA). Nonmitogenic doses of lipopolysaccharide (LPS) failed to show similar effects. Our data suggest that B cells at all levels of in vivo activation are responsive to stimulation by a growth factor present in EL4 supernatant, as manifested by cell growth and RNA synthesis. This activity has not previously been described for BCGF preparations. However, because the partially purified, EL4-derived supernatant used as BCGF in these studies has not been purified to homogeneity, we cannot conclude whether the factors that induce resting B cells to increase in size are the same as the growth factors that synergize with anti-mu to induce B cell proliferation or that maintain the proliferation of activated B cells.
Murine anti-swine T4 and T8 monoclonal antibodies: distribution and effects on proliferative and cytotoxic T cells.
Authors: Pescovitz M D , Lunney J K , Sachs D H
Published: December 2022

Two murine monoclonal antibodies reactive with porcine nylon wool-nonadherent T cells were examined by flow microfluorometry analysis for cellular distribution, and by functional analysis for their effect on MLR and CTL assays. The two determinants recognized, called PT4 and PT8, were shown to be expressed on almost mutually exclusive cell populations, with only 6 to 15% of T cells expressing both antigens. Among the 15% of nylon wool-nonadherent cells that did not express either PT4 or PT8, 5% were surface Ig+. In functional assays, cells that proliferated to PHA, hen egg lysozyme, and alloantigens were blocked by PT4 and were eliminated by PT4 and complement. PT4 and complement had no effect on CTL effectors or precursors. Reciprocally, PT8 blocked and PT8 and complement eliminated CTL effectors and precursors, but had no effect on proliferative responses. These results indicate that PT4 and PT8 define the porcine helper and cytotoxic T cell subsets, respectively.
The murine plasma cell antigen PC-1: purification and partial amino acid sequence.
Authors: Stearne P A , van Driel I R , Grego B , Simpson R J , Goding J W
Published: December 2022

The murine plasma cell alloantigen PC-1 is selectively expressed on B lymphocytes in their terminal phase of differentiation into antibody-secreting cells. Previous work on an analytical scale has shown that PC-1 consists of two apparently identical disulfide-bonded polypeptides, each of Mr 115,000. In this paper, we describe the generation of a monoclonal antibody to PC-1 and its use in the preparative isolation of PC-1 by affinity chromatography. Final purification to apparent homogeneity was achieved by preparative polyacrylamide gel electrophoresis. It was estimated that NS-1 myeloma cells possess 1 to 4 X 10(5) PC-1 monomers per cell on their surface. The yield of PC-1 after purification was approximately 10(5) monomers per cell. Purified PC-1 was digested with trypsin, and the resulting peptides were separated by reversed-phase high-performance liquid chromatography. Purified peptides were sequenced with a gas-phase sequencer.
Suppression of in vitro CML generation by alloactivated lymphocytes: analysis of antigen-nonspecific suppressive mechanisms.
Authors: Orosz C G , Ferguson R M
Published: December 2022

We have investigated the in vitro phenomena associated with antigen-nonspecific suppression of mixed lymphocyte culture (MLC) responses by allocativated lymphocytes. Using an experimental system that we described in a previous communication, we observed that a) the degree of suppressive activity generated by allocativation correlates directly with the intensity of proliferation observed during induction of suppressive activity, b) suppressive activity segregates exclusively with proliferating (lymphoblast) subpopulations of alloactivated lymphocytes, c) when suppressive cells are included in MLC, subsequent [3H]thymidine incorporation is enhanced and accelerated, rather than impaired, and d) a considerable proportion of the cells recovered from suppressed MLC appear to be the progeny of the suppressive population, and not the progeny of the MLC responder population. These data suggest that antigen-nonspecific suppression mediated by alloactivated lymphocytes has two related components: 1) cytokine preemption by suppressive (alloactivated) lymphocytes, and 2) MLC responder cell dilution by the progeny of suppressive lymphocytes. These data are consistent with the hypothesis that the antigen-nonspecific suppressive activity of alloactivated lymphocytes can reflect the coincidental ability of activated T cells to recognize and respond to mitogenic lymphokines in vitro. The data further explain why antigen-nonspecific suppression is difficult to reverse by addition of exogenous lymphokines to suppressed MLC.
Quantitative measurements of the specificity and kinetics of conjugate formation between cloned cytotoxic T lymphocytes and splenic target cells by dual parameter flow cytometry.
Authors: Perez P , Bluestone J A , Stephany D A , Segal D M
Published: December 2022

The formation of conjugates between cloned anti-H-2Kb and Dd cytotoxic T lymphocytes (CTL) and splenic target cells has been studied by dual parameter flow cytometry. By varying effector-target combinations and by blocking with anti-MHC class I monoclonal antibodies, we found that the specificity of conjugate formation, in general, paralleled that expected from cytotoxicity studies; however, a significant number of "nonspecific" conjugates was always observed. As expected from previous studies, conjugate formation did not occur below 10 degrees C and was inhibited by cytochalasin B, EDTA, and anti-Lyt-2 antibodies. Conjugate formation followed first-order kinetics. The rate of formation of conjugates increased with temperature from 24 degrees to 37 degrees C; at 37 degrees C, the half-time was 1.4 min. After a 6-min lag period, lysis of target cells could be detected at 37 degrees C but not at 30 degrees C or below. Because target cell lysis proceeded during a period of time when the number of conjugates remained constant, considerable effector cell recycling must have occurred. Comparisons of the fluorescence emissions from conjugated effector or target cells with those from unconjugated cells demonstrated that nearly all conjugates contained one effector cell and one target cell, independent of the ratio of the two cell types in the original mix. Once formed, anti-H-2Kb conjugates were stable when diluted into medium alone, but rapidly disaggregated in medium containing either anti-Lyt-2 or anti-Kb monoclonal antibodies, both of which blocked conjugate formation. This finding suggests that conjugates are normally stabilized by intercellular bonds that are constantly breaking and reforming at the cell:cell interface, and that the antibodies disrupt the conjugates by preventing the reformation of broken bonds.
Monoclonal antibodies against human mast cell tryptase demonstrate shared antigenic sites on subunits of tryptase and selective localization of the enzyme to mast cells.
Authors: Schwartz L B
Published: December 2022

Two murine monoclonal antibodies were prepared against tryptase, the major neutral protease and protein component of human mast cells. The antibodies were termed G5 (IgG2B-kappa) and H4 (IgG1-kappa). They were specific for tryptase by an enzyme-linked immunosorbent assay and an immunotransblot technique. The latter procedure showed that H4 and G5 each bind to the 35,000 and 37,000 m.w. subunits of tryptase, indicating immunologic cross-reactivity between the subunits. The monoclonal antibodies reacted only with tryptase subunits in an extract of dispersed lung cells. By immunofluorescence microscopy, tryptase was further identified to be present only in cytoplasmic granules of Alcian Blue-stained mast cells in dispersed pulmonary cell preparations. No evidence for a mast cell subtype lacking tryptase was detected. In addition, a procedure for the purification of tryptase to homogeneity from dispersed pulmonary cells containing less than 10% mast cells was developed; this procedure involved high salt extraction, ammonium sulfate precipitation, and sequential chromatography with decyl-agarose, DEAE-agarose, and heparin-agarose. The procedure resulted in a higher yield even with less pure starting material than reported previously. Tryptase is a selective marker for mast cells in dispersed pulmonary cells, and can be detected with specific anti-tryptase antibodies.
Comparison of MHC genes among distantly related members of the genus Mus.
Authors: Rogers M J , Germain R N , Hare J , Long E , Singer D S
Published: December 2022

The genomic content of class I and class II MHC DNA sequences in a variety of wild mice has been analyzed. The panel of mice includes members of three subgenera of the genus Mus. By genomic hybridization with the use of a variety of DNA probes, both class I and class II DNA sequences appear to be conserved in all of the species examined. However, the number of class I DNA sequences differs among the species. Furthermore, this variation appears to result from differential increases within subsets of class I genes. These data suggest that the class I multigene family is dynamic and changing over short periods of evolutionary time. In contrast, none of the class II genes appears to vary in copy number. More extensive polymorphism was noted amongst the class II beta genes than the alpha genes. Interestingly, the genomic sequence corresponding to E beta 2 is highly conserved, leading to the prediction that it is a genetically functional sequence.
Localization of the human MHC-linked complement genes between HLA-B and HLA-DR by using HLA mutant cell lines.
Authors: Whitehead A S , Colten H R , Chang C C , Demars R
Published: December 2022

A C4 cDNA clone has been used to localize the C4 genes within the human major histocompatibility complex (MHC). The clone, pC4AL1, detects a DNA polymorphism of the C4 genes in Southern blot hybridization experiments and established the genotype of a lymphoblastoid cell line, LCL 721, as heterozygous for the C4 DNA patterns. Subclones of LCL 721 that had gamma-ray-induced lesions of the MHC were analyzed with pC4AL1. Loss or retention of chromosome-specific C4 DNA patterns relative to the loss or retention of other MHC genes on the same chromosome was assessed. This permitted the mapping of the C4 genes, and the other MHC-linked complement genes, between the HLA-DR and HLA-B loci.
T cells with FC receptors in myeloma; suppression of growth and secretion of MOPC-315 by T alpha cells.
Authors: Müller S , Hoover R G
Published: December 2022

We have previously demonstrated that 1) BALB/c mice and patients with IgA myeloma developed a marked expansion of T cells with surface IgA-Fc receptors (T alpha cells), 2) the FcR alpha were induced by direct interaction of soluble myeloma IgA with T cells, and 3) the T alpha cells induced in IgA myeloma were Lyt-1-2+, IgA isotype-specific suppressor cells in normal immune responses. These findings established that the host with IgA myeloma responds to the large amounts of Ig produced by the tumor by activating an otherwise normal IgA isotype-specific suppressor circuit. In the present studies, we extend our previous observations and show that T alpha cells can suppress both the growth and secretion of MOPC-315 myeloma tumor cells. Thus, the isotype-specific immunoregulatory circuit activated in myeloma is capable of suppressing tumor cells as well as normal cells.
Blocking of CTL-mediated killing by monoclonal antibodies to LFA-1 and Lyt-2, 3. II. Evidence that trypsin pretreatment of target cells removes a non-H-2 molecule important in killing.
Authors: Gromkowski S H , Heagy W , Martz E
Published: December 2022

We sought additional evidence for an inverse relationship between functional CTL-target cell affinity on the one hand, and susceptibility of the CTL-mediated killing to inhibition by alpha LFA-1 and alpha Lyt-2,3 monoclonal antibodies on the other hand. Previously, we experimentally reduced affinity by pretreating the target cells with papain. This removed most of the class I H-2 antigens, had little effect on the ability of allospecific CTL to recognize and kill these targets, but dramatically reduced the initial strength of CTL-target cell adhesion, and increased by more than 10-fold the susceptibility of the killing to inhibition by alpha Lyt-2,3 and alpha LFA-1 MAb. In the present report, we find that pretreating the target cells with trypsin, like papain, does not significantly change the susceptibility of the target cells to killing by allospecific CTL in a 2-hr assay, and increases by about 10-fold susceptibility of the killing to inhibition by alpha LFA-1. Unlike papain, however, trypsin does not consistently increase blocking by alpha Lyt-2,3, does not remove class I H-2 antigens from the target cell, and does not substantially reduce the strength of initial CTL-target adhesion formation (estimated by post dispersion lysis after a 5-min conjugate-forming incubation). These results show a functional difference between LFA-1 and Lyt-2,3. Both papain and trypsin produced similar 10-fold increases in susceptibility to blocking by alpha LFA-1. In contrast, susceptibility to inhibition by alpha Lyt-2,3 was increased nearly 100-fold by papain, but was not consistently affected by trypsin. Thus, the above-mentioned inverse relationship holds for alpha Lyt-2,3 but not for alpha LFA-1. Our results are consistent with the hypothesis that Lyt-2,3 but not LFA-1 participates in recognition of class I H-2 antigens. Possibly LFA-1 participates in an adhesion-strengthening process that follows T cell recognition, and which may also be used by other LFA-1 expressing leucocytes in intercellular interactions. Finally, our results suggest (for the first time in the mouse system) that an unidentified non-H-2 "trypsin-sensitive counter blocking" molecule on the target cell plays an important role in CTL-target cell interaction.
Molecular species of platelet-activating factor generated by human neutrophils challenged with ionophore A23187.
Authors: Oda M , Satouchi K , Yasunaga K , Saito K
Published: December 2022

Two species of platelet-activating factor (PAF), 1-hexadecyl- and 1-octadecyl-2-acetyl-sn-glycero-3-phosphocholine (C16 = 0 AGEPC and C18 = 0 AGEPC) were detected in ionophore A23187-stimulated human neutrophils. The amount of AGEPC in 1 x 10(7) neutrophil cells was 80 +/- 26 pmol (mean +/- standard error) with a range of 14 to 223 pmol (n = 8), and it consisted of 80% of the C16 = 0 species and 20% of the C18 = 0 species. Most of the AGEPC derived from ionophore-treated neutrophils remained cell associated rather than being secreted into the medium, even when the medium contained ample albumin protein, which can trap AGEPC. These results were obtained by a technique of gas chromatography-mass spectrometry coupled with selected ion monitoring.
Effect of lipoxygenase metabolites of arachidonic acid on proliferation of human T cells and T cell subsets.
Authors: Gualde N , Atluru D , Goodwin J S
Published: December 2022

The lipoxygenase products LTB4 and 15 HPETE have been reported to stimulate T suppressor cell function and also to inhibit [3H]thymidine incorporation into mitogen-stimulated T cells. This present report documents that although these compounds do indeed inhibit [3H]thymidine incorporation into unfractionated T cells, they significantly enhance [3H]thymidine incorporation into T cell preparation enriched for cells bearing the cytotoxic suppressor cell phenotype identified by the OKT8 monoclonal antibody. The mitogen response of T cells enriched for OKT4+ helper-inducer cells is inhibited in manner similar to the response of unfractionated T cells. Thus, LTB4 and 15 HPETE stimulate both the function and the proliferation of the cytotoxic-suppressor T cell subset.
Stimulation of human neutrophilic granulocyte chemotaxis by monoclonal antibodies.
Authors: Laskin D L , Rovera G
Published: December 2022

Two mouse monoclonal antibodies, L12.2 and S5.22, were developed that are specific for human neutrophilic granulocytes and produce a twofold to threefold stimulation of n-formyl-methionine-leucyl-phenylalanine (FMLP)-induced chemotaxis. Stimulation of chemotaxis by the antibodies is specific for FMLP and is concentration dependent. L12.2 appears to be more potent in stimulating chemotaxis and is isotypically distinct from S5.22. In addition, although L12.2 reacts only with mature peripheral blood granulocytes, S5.22 reacts with leukemic cells of both myeloid and monocytic origin and with immature granulocyte precursor cells. This suggests that L12.2 interacts with an antigen that appears late in the differentiation pathway, whereas S5.22 binds to an antigen that is present throughout the myeloid lineage. By means of the under-agarose and Boyden chamber techniques, L12.2, but not S5.22, by itself was also found to be a potent granulocyte chemoattractant. Cells in a gradient of L12.2 display polarized and oriented morphology. L12.2 alone, but not S5.22, also stimulates granulocyte phagocytosis and induces superoxide anion production. Neither L12.2 nor S5.22 affected the release of myeloperoxidase or lysozyme from granulocytes either alone or in combination with FMLP, C5a, or the tumor promoter, 12-O-tetradecanoyl-phorbol-13-acetate (TPA). These results suggest that L12.2 interacts with a single antigenic determinant on granulocytes that is involved in chemotaxis, phagocytosis, and superoxide anion release.
Rejection of ticks from guinea pigs by anti-hapten-antibody-mediated degranulation of basophils at cutaneous basophil hypersensitivity sites: role of mediators other than histamine.
Authors: Brown S J , Askenase P W
Published: December 2022

Previous studies have established that recruitment of basophils to sites of tick feeding in guinea pigs is required to effect immune resistance. In the current study, actively sensitized guinea pigs treated three times daily with H-1 (mepyramine) and H-2 (cimetidine) histamine receptor antagonists, during the challenge tick infestation period, expressed normal resistance to Amblyomma americanum larvae. Similarly, naive guinea pigs treated with anti-histamines four times daily, beginning 7 days before transfer of immune serum and tick challenge and continuing through the tick infestation period, also expressed normal antibody-mediated resistance to A. americanum. These results indicated that histamine was not an important basophil mediator of the resistance response. Ticks allowed to feed on tissue rich in basophils that were induced by sensitization and subsequent local challenge with non-tick protein antigen, keyhole limpet hemocyanin (KLH), expressed normal yield. Ticks that fed on similar tissue rich in basophils induced by sensitization and challenge with KLH, in which the basophils expressed anti-picryl specificity due to systemic passive transfer of anti-picryl antibodies, were rejected when basophils were induced to degranulate by i.v. challenge with picryl antigen at 6 hr (29% rejection), 12 hr (18% rejection), 24 hr (22% rejection), and 48 hr (37% rejection) post-tick attachment. However, basophil degranulation at 18, 72 and 96 hr post-tick attachment had no adverse effect on tick feeding. These hosts were protected from systemic anaphylaxis by treatment with the anti-histamine mepyramine. Release of histamine occurred at tick feeding sites, but vasoactive effects were blocked by mepyramine treatment as evidenced by a lack of increased vascular permeability (bluing) at these sites compared with non-tick-infested tissues, or to cutaneous basophil hypersensitivity (CBH) sites of animals not protected with mepyramine. These results indicate that local recruitment and subsequent degranulation of basophils via immune mechanisms dependent on non-tick antigens can lead to tick rejection, and that basophil-derived mediators other than histamine are involved in this immune resistance response to A. americanum ticks. The identity of the crucial basophil mediator(s) is not known. The significant susceptibility of ticks to basophil-mediator release at 6 to 12 hr and 24 to 48 hr post-attachment coincides with the tick attaching and fast-feeding phases, respectively, suggesting that these phases of tick parasitism are particularly susceptible to the effect of basophil mediators other than histamine.
Filarial parasite-specific T cell lines: induction of IgE synthesis.
Authors: Nutman T B , Volkman D J , Hussain R , Fauci A S , Ottesen E A
Published: December 2022

The development of T lymphocyte lines and clones of defined specificity has provided an important method for investigating T cell recognition of foreign antigens as well as T cell influence on B cell activity. We described previously a parasite-specific T cell line (TCL) derived from a patient with a naturally acquired filarial infection and elevated levels of serum IgE. The TCL is composed of Leu-3+ helper cells and is maintained independent of exogenous growth factors. In the present study, we used these T cells to investigate their immunoregulatory function on the in vitro IgE response. These parasite-specific T cells can provide isotype-specific help for antigen-induced IgE production by B cells in vitro. Autologous T cells profoundly suppress IgE production in a concentration-dependent manner. Furthermore, soluble factors generated from these filarial-specific TCL after antigen stimulation are able to induce the production of IgE in normal human cells not already synthesizing measurable amounts of IgE in vitro. Partial physicochemical characterization of this factor has shown that it is heat labile, has an m.w. between 10,000 and 30,000 M(r), and is a mannose-rich glycoprotein.
Induction of neutralizing antibody in mice against poliovirus type II with monoclonal anti-idiotypic antibody.
Authors: Uytdehaag F G , Osterhaus A D
Published: December 2022

Syngeneic monoclonal anti-idiotope antibody Ab2,2-17C3SCC was raised against an idiotope on a protective monoclonal antibody with specificity for poliovirus type II. Ab2,2-17C3SCC detects a paratope-related interspecies IdX. Ab2,2-17C3SCC purified from supernatant fluids of hybridoma cells by protein A-Sepharose was injected into 4- to 6-wk-old BALB/c mice. The sera of the mice were screened for the expression of antibodies bearing the corresponding idiotope. Immunization of mice with Ab2,2-17C3SCC induced antibodies of complementary specificity. Furthermore, micro VN tests suggest that Ab2,2-17C3SCC can substitute for antigen in the induction of anti-polio neutralizing antibodies, and hence can function as a monoclonal anti-idiotypic vaccine.
Identification of several cell surface proteins of non-T, non-B acute lymphoblastic leukemia by using monoclonal antibodies.
Authors: Quackenbush E J , Letarte M
Published: December 2022

Monoclonal antibodies (MAb) were produced by immunization of BALB/c mice with cells from a non-T, non-B acute lymphoblastic leukemia (ALL) cell line. Nine distinct antigens (groups I to IX) were defined by these monoclonal antibodies, some of which appear to be associated with specific stages of cellular differentiation. The number of molecules of each MAb reactive with the ALL cell line, measured in a quantitative cellular radioimmunoassay, varied from 0.6 X 10(5) to 11 X 10(5) molecules/cell, indicating that the antigens identified represent major constituents of the cell surface. The biochemical nature of the antigens was examined on the ALL cell line by antibody affinity chromatography and/or immunoprecipitation and SDS-polyacrylamide gel electrophoresis. Groups I through III are composed of previously described antigens: HLA class I, HLA class II molecules, and CALLA, the common ALL antigen. The other MAb define antigens previously undescribed on non-T, non-B ALL cells. Group IV antigen is a polypeptide of apparent m.w. 95,000 distinct from CALLA. It is expressed on some ALL samples and on the vascular endothelial cells of several tissues. Group V antigen is a single polypeptide chain of m.w. 94,000, also distinct from CALLA and expressed by lymphocytes, thymocytes, acute myelogenous leukemia (AML) cells, and ALL cells. Group VI is a molecular complex composed of two noncovalently associated polypeptides of apparent m.w. 125,000 and 87,000 and appears to be restricted to ALL, AML, macrophages, and hematopoietic precursor cells. Group VII is a glycoprotein of apparent m.w. 85,000, which, within the thymus, is primarily restricted to the medullary area. It is also present on AML, bone marrow cells, and mature T and B lymphocytes. Group VIII is a disulfide-linked complex of apparent m.w. greater than 120,000 under nonreducing conditions. It is resolved into three major polypeptides of apparent m.w. 57,000, 47,000, and 41,000 under reducing conditions. This complex is found in greatest amounts on the non-T, non-B ALL cell line but is also present on AML, ALL, and on subpopulations of normal bone marrow and tonsil cells. Group IX antigen is a single polypeptide chain of apparent m.w. 51,000 on the ALL cell line. This antigen is expressed strongly on ALL and AML samples and on normal bone marrow; much lower antigenic density is found on thymus and tonsil cells. The antigens described here with a series of MAb produced in a single fusion represent a unique array of cell surface molecules of non-T, non-B ALL cells.(ABSTRACT TRUNCATED AT 400 WORDS)
Specific effect of anti-transferrin antibodies on natural killer cells directed against tumor cells. Evidence for the transferrin receptor being one of the target structures recognized by NK cells.
Authors: Alarcón B , Fresno M
Published: December 2022

Treatment of PBL or Percoll-isolated LGL with anti-transferrin antibodies plus complement reduced their natural killing activity against K-562 cells between 30 and 70%. The same antibodies inhibited natural cytotoxicity when added directly to the assay. Similar depletion or inhibition of NK cytotoxicity was observed when using HeLa cells as targets. The decrease or inhibition by transferrin antibodies was less marked when IFN-treated PBL or LGL as effector cells were used. The inhibition of anti-transferrin antibodies seems to be located at the level of the effector cell population. When PBL but not target K-562 cells were pretreated with anti-transferrin antibodies and were washed before use in the assay, cytotoxicity was decreased by 50%. In addition, about 80% of the LGL positively selected on anti-transferrin plates stained with Leu-11. Furthermore, no reduction by anti-transferrin antibodies plus complement treatment of PBL or LGL, or inhibition by antibodies alone, was observed when the cells were tested against HSV-1-infected cells. Membrane extracts from LGL inhibited NK cytotoxicity against K-562 or HeLa cells. Moreover, the inhibitory component of this extract was removed by anti-transferrin IgG but not by control IgG. These results are in agreement with the recent hypothesis that NK cells recognize the transferrin receptor in tumor target cells, because both the transferrin receptor and anti-transferrin antibodies may share a similar structure that interacts with the NK cells.
Functional heterogeneity in allospecific cytotoxic T lymphocyte clones. II. Development of syngeneic cytotoxicity in the absence of specific antigenic stimulation.
Authors: Claesson M H , Miller R G
Published: December 2022

Two out of four long-term murine allospecific cytotoxic T lymphocyte (CTL) clones tested could develop high levels of cytotoxicity against syngeneic target cells when cultured under appropriate conditions. All CTL clones maintained strict allospecificity so long as they were cultured with both appropriate allogeneic stimulator cells and growth factor (supernatant from secondary mixed lymphocyte cultures). In two of the clones, syngeneic reactivity rapidly developed when the allogeneic stimulator cells were replaced with syngeneic or third party stimulator cells, and when the supernatant from EL4 thymoma cells stimulated with phorbol ester was used as growth factor. In addition to killing the appropriate allogeneic target, clones with syngeneic reactivity could kill both syngeneic C57BL/6 targets and H-2-congenic BALB.B targets but not third party unrelated targets, suggesting that the self structure recognized was coded for within the major histocompatibility complex. Such clones did not kill the natural killer (NK) target YAC. The results obtained from cold target inhibition and from subcloning at limiting dilution of clones with syngeneic reactivity suggested that both allogeneic and syngeneic reactivity could be expressed by the same individual cell in the CTL clone. The specificity for syngeneic H-2 as opposed to third party H-2 and NK-sensitive target cells, and the observation that both allospecific and syngeneic killing could be partially blocked by anti-Lyt-2 antibody treatment of the CTL, strongly suggested that different recognition structures are involved in CTL-mediated syngeneic cytotoxicity and NK cytotoxicity.
Induction of proliferation in vitro of resting human natural killer cells: expression of surface activation antigens.
Authors: London L , Perussia B , Trinchieri G
Published: December 2022

In this report, we show that resting human peripheral blood NK cells, positively enriched with antibody B73.1, can be induced to proliferate in vitro in short-term cultures, and newly express membrane activation antigens. B73.1+ NK cells, cultured for 6 days in the presence of conditioned medium containing IL 2 and irradiated B lymphoblastoid cells, show significant [3H]TdR incorporation beginning on day 4. At that time, a large proportion of the cells express HLA-DR and 4F2 antigens, and transferrin and IL 2 receptors, all detectable at high density by indirect immunofluorescence with the use of monoclonal antibodies. These cells maintain their ability to lyse target cells spontaneously or in the presence of antibodies. By two-color immunofluorescence and cell cycle analysis combined with indirect immunofluorescence, we demonstrate directly that the activation antigens are expressed on all NK (B73.1+) cells in the S/G2/M phases of the cell cycle, and on only a proportion of those in the G0/G1 phases.