bluuebunny/crossref_metadata_2025 huggingface dataset:
Immune and enzyme histochemical studies of a human hepatocellular carcinoma cell line producing hepatitis B surface antigen.
Authors: Gerber M A , Garfinkel E , Hirschman S Z , Thung S N , Panagiotatos T
Published: December 2022

PLC/PRF/5, a tissue culture cell line derived from a human hepatocellular carcinoma and producing hepatitis B surface antigen (HBsAg), was studied by immune and enzyme histochemical techniques. HBsAg was demonstrated in the cytoplasm and on the surface of tumor cells. The percentage of HBsAg-positive cells in subculture increased with time until almost all cells expressed HBsAg when the monolayer reached confluence. Similar patterns were found for alpha 1-anti-trypsin and carcino-embryonic antigen, whereas alpha-fetoprotein was observed only in small foci of cells. Hepatitis B core antigen and albumin were not detected. gamma-Glutamyl transferase activity was markedly increased in the tumor cells, whereas adenosine triphosphatase and glucose-6-phosphatase activities were not demonstrable. Patterns of antigenic expression and enzyme phenotype of PLC/PRF/5 cells show remarkable resemblance to those observed in vivo in human hepatocellular carcinoma. Therefore, this cell line may be a useful model to study the control and modulation of both oncofetal antigens and HBsAg.
A comparison of the suppressor cells found in the spleens of 89SR-treated mice and in normal murine bone marrow.
Authors: Levy E M , Corvese J S , Bennett M
Published: December 2022

Normal murine bone marrow cells and spleen cells of mice treated with 89Sr both have suppressive activity. These nonspecific suppressor cells inhibit the ability of normal spleen cells to undergo antibody responses in vitro. After being precultured for 24 hr, these cells will also suppress antibody responses in vivo and the responses of normal spleen cells to T and B cell mitogens in vitro. These cells have previously been shown not to be mature T or B lymphocytes or macrophages. Velocity sedimentation and cell-size analysis indicated that both suppressor cells are large (approximately 206 mu3). Mitomycin C treatment eliminated the ability of both suppressor cells to inhibit an in vitro antibody response. In contrast, this treatment did not reduce the ability of the cells to suppress a mitogenic response. Irradiation (1000 R) was also ineffective in eliminating the ability of either cell to suppress a mitogenic response We conclude that the 2 suppressor cells are closely related if not identical, and we speculate that these cells may function in vivo to suppress immune reactivity in areas of intense hematopoiesis.
Membrane protein synthesis in mitogen-stimulated human T lymphocytes.
Authors: Udey M C , Parker C W
Published: December 2022

Proteins were biosynthetically labeled with [35S]-methionine in resting and mitogen-stimulated human peripheral blood lymphocytes. Lymphocytes were disrupted by sonication, the membrane fraction was isolated by differential centrifugation, and radiolabeled membrane proteins were identified after 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and autoradiography. The T cell mitogen concanavalin A (Con A) produced selective increases in the incorporation of [35S]-methionine into a membrane protein (or family of 4 structurally related membrane proteins) with an apparent m.w. of 28,000 and approximate isoelectric points (pI) of 5.5, 5.7, 6.0, and 6.2 (p28). Quantitative microdensitometry was used to evaluate p28 synthesis in a large number of different lymphocyte donors. Consistent and highly significant differences between resting and stimulated cells were demonstrable whether the 4 p28 spots were considered individually (p less than 0.0025) or in sum (p less than 0.0005). The p28 response to Con A was shown to be contained within the T lymphocyte fraction of mononuclear cells, was dose dependent over the mitogenic concentration range for Con A, and was blocked by alpha-methyl-D-mannoside, a sugar inhibitor of Con A binding. Differences between stimulated and resting cells were readily apparent within 2 hr of activation and became progressively larger with time. Protein p28 was prominently represented in plasma membranes isolated on discontinuous sucrose density gradients and was identified as a cell surface glycoprotein based on studies in which intact stimulated cells were exposed to degradative enzymes. This protein was lost from cells treated with pronase, whereas neuraminidase digestion changed the pI of p28 from a range of 5.5 to 6.2 to a range of 6.1 to 6.4 without changing the apparent m.w.
Human T cell colony formation in microculture: analysis of growth requirements and functional activities.
Authors: Gelfand E W , Lee J W , Dosch H M , Price G B
Published: December 2022

A microculture method in methylcellulose has been developed for the study of human T cell colony formation. The technique is simple, reliable, does not require preincubation with lectin and requires small numbers of cells. Colony formation was dependent on the presence of phytohemagglutin-conditioned medium, a T colony precursor cell (TCPC), and a "helper" or accessory T cell. Plating efficiency was increased 10-fold in the presence of irradiated feeder cells. Progenitors of the T colony cells were identified in peripheral blood, tonsil, and spleen but not in thymus or thoracic duct. They were isolated in the E-rosetting, theophylline-resistant, Fc-IgG-negative cell populations. In peripheral blood the frequency of TCPC and accessory cells, the T colony forming unit, was estimated to be 8 X 10(-3). Colony cells proliferated in response to lectins and allogeneic cells. Forty to 80% of the cells were Ia-positive and stimulated both autologous and allogeneic mixed lymphocyte responses. They were incapable of mediating antibody-dependent cytotoxicity. In contrast, they were effective in assays of spontaneous cytotoxicity but only against certain target cells. This method for the analysis of T colony formation should prove valuable in the functional analysis of T cell subsets in immunodeficiency states or the transplant recipient.
Suppression of the mixed leukocyte response and of graft-vs-host disease by spleen cells following total lymphoid irradiation (TLI).
Authors: King D P , Srober S , Kaplan H S
Published: December 2022

Total lymphoid irradiation (TLI) was administered to mice as 17 fractions of 200 rads delivered to the major lymphoid organs. Spleen cells capable of suppressing the in vitro mixed leukocyte response (MLR) and in vivo graft-vs-host disease (GVHD) were found in mice after treatment with TLI. Suppression was not antigen specific and was markedly reduced by treatment of the spleen cells with anti-Thy-1.2 antiserum and complement. Suppressor activity declined with time after irradiation and disappeared within 30 to 40 days. The evidence suggests that the suppressor cells may prevent initial BM rejection and acute GVHD in allogeneic BM transplant recipients prepared with TLI.
Identification of types of cells in human peripheral blood that bind C1q.
Authors: Tenner A J , Cooper N R
Published: December 2022

Earlier studies showed that approximately 26% of the cells present in human mononuclear cell preparations had the ability to bind purified monomeric C1q. The present studies were initiated to identify the cell types comprising the C1q binding population. Double marker fluorescence, rosetting, and morphologic studies on cell preparations depleted of or enriched in various cell types were simultaneously employed to identify those subpopulations that bound C1q. C1q binding was detected by fluorescent techniques (with FI-F(ab')2 anti-C1q). Monocytes in mononuclear cell preparations were detected by the ability to phagocytose carbonyl iron. B cells were identified by reactivity with rhodamine-conjugated F(ab')2 anti-human F(ab')2 and by rosetting with erythrocytes bearing C3b. These studies showed that monocytes and B lymphocytes comprised the majority of C1q-binding cells in mononuclear cell preparations, whereas T lymphocytes lacked this property. In addition, a minor population of nonphagocytic cells in such preparations that lacked B and T cell markers also bound C1q. Finally, a high but variable proportion of polymorphonuclear leukocytes bound C1q. Binding of C1q to PMN was concentration-dependent, saturable and specific and exhibited an equilibrium constant of 0.76 X 10(7) M-1. Thus, PMN also possess a specific receptor for C1q.
Molecular basis for the temperature-dependent insolubility of cryoglobulins. X. The amino acid sequence of the heavy chain variable region of McE.
Authors: Gerber-Jenson B , Kazin A , Kehoe J M , Scheffel C , Erickson B W , Litman G W
Published: December 2022

The amino acid sequence of the VH region of McE, a monoclonal IgM cryoimmunoglobulin, has been determined employing automated sequencing methodology. Digestion of the intact Fab mu component, derived by trypsin cleavage of the parent protein at elevated temperature with CNBr, followed by complete reduction and alkylation, yielded the intact light chain as well as the 2 CNBr fragments that constituted the VH. N-terminal sequencing of the larger unblocked CNBr fragment, along with a component fragment derived by cleavage by BNPS-Skatole, established the structure of the VH from position 88 through the V leads to C switch region. Citraconylation of the smaller, blocked fragment effected sufficient solubilization for enzymatic deblocking and direct sequencing of the N-terminal 20 residues of the VH. Complete trypsin digestion of the N-terminal CNBr fragment yielded 9 peptides that could be isolated by preparative cation exchange chromatography and gel filtration. The complete sequence of these peptides along with 4 chymotryptic peptides completed the primary structure of the VH region. The primary structure of McE appears to resemble that of He, previously identified as belonging to the VH II subgroup. The presence of characteristic CDR and FR regions as well as the identification of a probable site of glycosylation suggest that the cryoimmunoglobulin closely resembles noncryoglobulins in terms of overall structural composition. The cryoglobulin property may arise through alterations in individual residues or unfavorable arrangements of CDR and FR segments.
Human thy-1: unusual localization and possible functional significance in lymphoid tissues.
Authors: McKenzie J L , Fabre J W
Published: December 2022

BALB/c mice were immunized with purified human brain Thy-1 and used in cell fusion experiments to derive an anti-human Thy-1 monoclonal antibody. Specificity was proven by a) showing that the antigen recognized had precisely the tissue distribution expected of human Thy-1 from previous studies, and b) demonstrating that the molecule purified from human brain by monoclonal antibody affinity columns 1) had the same mobility on SDS PAGE as pure rat Thy-1 and 2) could inhibit an assay previously shown to be directed at the human-rat cross-reactive component of Thy-1. Studies with the fluorescence-activated cell sorter confirmed that Thy-1 is absent from human blood lymphocytes and cell suspensions from human lymphoid organs, except that very weak fluorescence could be detected on 7% of thymus cells in suspension. However, fluorescence studies on frozen sections showed bright staining restricted mainly to the periphery of the thymus lobule, the marginal zone and some periarteriolar lymphocytes in the spleen, and the post-capillary venules of lymph node. In some lymph nodes, a halo of Thy-1 positive cells was seen around lymphatic nodules or germinal centers, but this was not a constant finding. This distribution is different from the known distribution of Thy-1 in the lymphoid tissues of the mouse and rat. Our studies suggest that human Thy-1 might be involved in lymphocyte recirculation, and that it is a marker for early T lymphocytes in man. The studies also show that the few Thy-1 positive cells in human lymphoid organs are selectively lost in the preparation of single-cell suspensions.
Age-associated increase in expression of the T cell surface markers Thy-1, Lyt-1, and Lyt-2 in congenitally athymic (nu/nu) mice: analysis by flow microfluorometry.
Authors: MacDonald H R , Lees R K , Sordat B , Zaech P , Maryanski J L , Bron C
Published: December 2022

The expression of T cell-associated surface markers by lymphoid cells from congenitally athymic (nude) mice has been quantitatively investigated using flow microfluorometry. Spleen and lymph nodes from old (greater than 6 mo) nude mice on either a C57BL/6 or BALB/c genetic background were found to contain significant numbers (5 to 13% in spleen and 15 to 24% in lymph nodes) of cells expressing Thy-1 antigen. The proportion of Thy-1 positive cells in nude spleen was dramatically increased (to 22 to 67%) after passage of the cells over nylon wool columns. In contrast to older animals, young (1- to 2-mo-old) nude mice had undetectable levels (less than 1%) of Thy-1 bearing cells in spleen and reduced levels (6%) in lymph nodes. After passage of their spleen cells over nylon wool, some Thy-1 positive cells (10%) were detectable in 2-mo-old nude mice but none were detectable at 1 mo. In addition to Thy-1, we were able to detect the T cell alloantigens Lyt-1 and Lyt-2 on nylon wool-passed spleen cells from older C57BL/6 or BALB/c nu/nu mice. In general, the proportion of Lyt-1 bearing cells in nude lymphoid populations was similar to the proportion of Thy-1 positive cells. A smaller fraction of nude cells (corresponding to 35 to 59% of the total Thy-1 positive cells) were found to express Lyt-2. Analysis of the forward light scatter distributions of nude lymphoid cells bearing either Thy-1, Lyt-1, or Lyt-2 antigens further demonstrated that an overlapping population of relatively large-size cells expressed these surface markers. These data strongly imply that at least 2 subsets of T cells (i.e., Lyt-1+2- and Lyt-1+2+) develop in older nude mice in the apparent absence of thymic influence.
Accessory cells in the in vitro generation of type C viruses specific T killer lymphocytes. I. Role of macrophages in primary anti-FMR reaction.
Authors: Gomard E , Wybier-Franqui J , Levy J P
Published: December 2022

The role of macrophages has been studied in in vitro cytolytic T lymphocytes- (CTL) mediated responses directed against the FMR cell-surface antigens induced by C type viruses. Macrophages, defined as Thy 1.2-negative, Ia-positive, adherent, phagocytic, radioresistant cells present in the spleen and the peritoneal cavity, are required to obtain primary in vitro anti-FMR responses. In most of our experiments they were not necessary in secondary reactions. In primary responses, macrophages and responder T cells must be compatible at least at the I region of the major histocompatibility complex. Anti-Ia antibodies inhibit the response. A rat soluble factor (Interleukin 2) can replace macrophages in primary anti-FMR CTL-mediated reactions. These results suggest that macrophages function during primary anti-FMR response by interacting with a helper T cell rather than with CTL precursors. In agreement with this hypothesis, it appears that the H-2 restriction of F1 hybrid anti-FMR CTL generated in vitro in primary reaction is not related to the H-2 specificities of the parental macrophages used and depends only on the H-2 antigens of the stimulating tumor cells.
Tuftsin: a naturally occurring immunopotentiating factor. I. In vitro enhancement of murine natural cell-mediated cytotoxicity.
Authors: Phillips J H , Babcock G F , Nishioka K
Published: December 2022

Tuftsin is a physiologic tetrapeptide, which has recently been shown to possess immunoadjuvant properties including the stimulation of macrophage and granulocyte phagocytosis, migration, bactericidal, and tumoricidal activities. Tuftsin has also been reported to possess in vivo immunologically mediated anti-tumor potential. To determine the potential role of tuftsin as an antineoplastic immunoadjuvant, the in vitro effects of tuftsin on murine natural cell-mediated cytotoxicity were studied. We observed that in vitro treatment of mouse splenic effector cells with synthetic tuftsin induced a pronounced enhancement of natural killer cell (NKC) cytotoxicity against the T cell lymphoma Yac-1. The magnitude of NKC enhancement was directly dependent upon the concentration of tuftsin employed, with maximum NKC stimulation observed at tuftsin concentrations of 50 to 100 microgram/ml. The tuftsin induced enhancement of NKC activity was not strain specific, since equivalent stimulation was seen in CBA/J, C56BL/10, and DBA/2 mice. Elimination of macrophages, monocytes, T cells, and immunoglobulin-bearing cells had no effect on the dose-dependent tuftsin stimulation of natural cell-mediated cytotoxicity; thus the characteristics of the effector cells activated by tuftsin were consistent with those reported for NKC. We also observed that treatment of splenic effector cells with tuftsin prolonged the cytotoxic capabilities of these cells beyond 18 hr.
Induction of IgG memory responses with polyvinylpyrrolidone (PVP) is antigen dose dependent.
Authors: Lite H S , Braley-Mullen H
Published: December 2022

Irradiated recipients of spleen cells from mice primed with a very low dose (0.0025 microgram) of the thymus-independent (TI) antigen polyvinylpyrrolidone (PVP) produced PVP-specific IgG memory responses after secondary challenge with a T-dependent (TD) form of PVP, PVP-HRBC. The IgG memory responses induced by low doses of PVP were similar in magnitude to those induced by the TD antigen PVP-HRBC. The induction of IgG memory by the TI form of antigen was markedly dependent on the dose of PVP used to prime donor mice. Spleen cells from mice primed with an amount of PVP (0.25 microgram) that induces an optimal primary IgM response did not produce significant IgG antibody after challenge with PVP-HRBC. The inability of higher doses of PVP to induce IgG memory may be due, at least in part, to the fact that such doses of PVP were found to induce tolerance in PVP-specific B cells and could suppress the induction of memory induced by PVP-HRBC. Low doses of PVP did not interfere with the induction of memory by PVP-HRBC. Expression of IgG memory responses in recipients of PVP-HRBC or low-dose PVP-primed cells was found to be T cell dependent. Moreover, only primed T cells could reconstitute the response of recipients of primed B cells, suggesting that the ability of PVP to induce IgG memory may be related to its ability to prime T helper cells. Expression of the IgG memory response in recipient mice also required the use of a TD antigen for secondary challenge, i.e., mice challenged with PVP did not develop IgG. The inability of PVP to elicit IgG antibody was due in part to the fact that 0.25 microgram PVP was tolerogenic, i.e., it could interfere with the expression of memory in mice simultaneously challenged with both PVP and PVP-HRBC.
Identification of H-2-controlled structural variants of the murine Slp protein and demonstration of cis-regulation of its expression.
Authors: Parker K L , Carroll M C , Shreffler D C , Capra J D
Published: December 2022

H-2 haplotype-related structural variation in Slp beta-chains was detected by the limited proteolysis peptide mapping technique. Two distinct peptide patterns were distinguished: Slp.1 was found in strains carrying the S regions of the H-2d and H-2s haplotypes, and Slp.2 was found in strains bearing the S region of H-2w7. These different patterns were expressed codominantly in male (Sd X Sw7)F1 hybrids, whereas only the Slp.2 pattern was expressed in female heterozygotes. The 2 beta-chains are most likely the products of alleles of the Slp structural gene, which must then be located in the murine major histocompatibility complex, very probably in the S region.
Alterations of interferon production in a mouse model of thermal injury.
Authors: Suzuki F , Pollard R B
Published: December 2022

The effect of thermal injury on the response of interferon (IFN) production in vivo and in vitro after stimulation with eight representative inducers was investigated in a mouse model. The response of mice to immune IFN (IFN-gamma) inducers, staphylococcal enterotoxin A, concanavalin A, and a specific antigen for BCG-sensitized lymphocytes (purified protein derivative) was impaired after a 30% total body surface area third-degree burn. Suppression of IFN-gamma production was observed at day 2 and persisted until day 7 after burn. Decreased IFN-gamma production correlated closely with the percentage of total body surface area burned. When virus type IFN (IFN-alpha/beta) inducers, Newcastle disease virus, polyriboinosinic-polyribocytidylic acid, 10-carboxymethyl-9-acridanone, and E. coli endotoxin, were administered to mice, no change in IFN response was observed after thermal injury. Similar results were obtained when spleen cells obtained from thermally injured mice were stimulated with IFN-gamma inducers in vitro. These studies suggest that although the capacity for IFN-alpha/beta production remains intact in thermally injured mice, IFN-gamma production may be selectively decreased in burned animals and in their spleen cells.
Mechanism for the suppression of gamma-interferon responsiveness in mice after thermal injury.
Authors: Suzuki F , Pollard R B
Published: December 2022

As reported previously, gamma-interferon production was decreased after the administration of inducers to thermally injured mice as compared with noninjured controls. Similarly, spleen cells from injured mice had decreased ability to produce interferon in vitro after stimulation with inducers. The present study demonstrated that the decrease in interferon production was associated with the presence of suppressor cells in the spleen of burned mice that were capable of inhibiting interferon production by normal splenic lymphocytes in vitro. Passive transfer of spleen cells containing suppressor cell activity derived from injured mice induced suppression in normal mice, and the time of the appearance of suppressor cell activity in injured mouse spleens closely approximated the time of the appearance of the suppression of interferon production observed in mice after thermal injury. The suppressor cells were characterized as a population of macrophages by the following: they adhered to plastic surface and could be removed from spleen cells by carbonyl-iron treatment; treatment of plastic-adherent cells with anti-Thy-1.2 and anti-mouse immunoglobulin antisera followed by complement failed to abrogate the suppression produced by these cells.
Cell surface alpha 1-protease inhibitor on human peripheral mononuclear cells in culture.
Authors: Boldt D H , Chan S K , Keaton K
Published: December 2022

We have studied expression of alpha 1-protease inhibitor (alpha 1-PI) by human mononuclear cells. alpha 1-PI was detected on 50% of freshly isolated peripheral mononuclear cells. Unless a proliferative stimulus was provided, alpha 1-PI subsequently disappeared from the cell surfaces. Plant mitogens, periodate, neuraminidase-galactose oxidase, or allogeneic cells all were effective stimuli of alpha 1-PI expression. Concanavalin A stimulated de novo synthesis of alpha 1-PI in cell cultures containing both lymphocytes and mononuclear phagocytes, and alpha 1-PI simultaneously appeared on surfaces of activated lymphocytes. Inhibition of protein synthesis by cycloheximide or monocyte depletion abolished de novo alpha 1-PI synthesis, but only monocyte depletion inhibited alpha 1-PI expression. Lymphocytes, but not monocytes, displayed saturable binding of radioiodinated alpha 1-PI. The data are consistent with the interpretation that human mononuclear phagocytes synthesize and secrete alpha 1-PI. When protein synthesis is inhibited, mitogenic stimuli may provoke release of previously synthesized alpha 1-PI from mononuclear phagocytes. Secreted alpha 1-PI then may bind to specific lymphocyte cell surface receptors. This pattern of alpha 1-PI synthesis, secretion, binding, and expression on lymphoid cell surfaces appears to be a common characteristic of many immunologic reactions in vitro.
Analysis of cloned T cell function. I. Dissection of cloned T cell proliferative responses using cyclosporin A.
Authors: Orosz C G , Fidelus R K , Roopenian D C , Widmer M B , Ferguson R M , Bach F H
Published: December 2022

CsA interferes in a specific manner with the expansion of T cell clones in that it inhibits the antigen-driven component of the proliferative responses made by cloned helper T cells, cloned conventional cytolytic T cells, and cloned helper-independent cytolytic T cells. Cloned helper T cells and helper-independent cytolytic T cells, which share the ability to proliferate when cultured with specific alloantigen, fail to proliferate when cultured with specific alloantigen, fail to proliferate in response to this stimulus in the presence of CsA (10 to 100 ng/ml). In contrast, the proliferation observed when these cells are cultured with exogenous growth factors (but not alloantigen) is little influenced by as much as 1000 ng/ml CsA. When cloned helper T cells or helper-independent cytotoxic T cells are cultured with alloantigen plus exogenous growth factor, additive or synergistic proliferation occurs. However, CsA (10 to 1000 ng/ml) blocks only the component of proliferation induced by alloantigen, and leaves the lymphokine-driven component intact. CsA has similar effects on the proliferation of cloned conventional cytolytic T cells. Thus, CsA separates cloned T cell proliferation into two components: one driven by contact with alloantigens, the other driven by contact with mitogenic lymphokines.
In vivo allogeneic effects: shift in the isotype profile of primary TI-2 responses in mice undergoing graft-vs-host reaction.
Authors: Golding H , Rittenberg M B
Published: December 2022

We showed previously that primary responses to T-dependent (TD) and T-independent type 2 (TI-2) antigens were differentially affected by allogeneic effects induced in vivo during a graft-vs-host reaction (GVH). TD responses were greater than or equal to 80% suppressed, whereas the TI-2 responses were greatly enhanced, particularly the IgG component, which normally is very low. We have analyzed the IgG subclass distribution in primary responses of normal and GVH F1 mice in order to determine whether the strong T cell signals that occur during GVH reactions also induce shifts in the isotype profile. The effect of GVH on responses to TI-2 antigens was of particular interest because they are usually dominated by IgM and IgG3 classes in normal mice. We found a threefold to 10-fold increase in the PFC numbers of all four IgG subclasses in the response to TI-2 antigens, with an apparent shift from the usual IgG3 dominance to IgG1 in GVH mice. This IgG1 dominance was not found in serum antibodies where IgG3, IgG1, and IgG2b were equally expressed, although total IgG was increased greater than 20-fold. No isotype shift was found in either the TNP-KLH response, which was greater than or equal to 75% suppressed (IgG1 dominance was retained), or in the TI-1 response to TNP-Ba. The latter response was reduced (25 to 50%) in GVH mice and continued to be dominated by IgG2b/2a and IgG3. Unlike the unique isotype patterns found in primary responses, TNP-KLH primed mice challenged with TD, TI-1, or TI-2 antigens gave memory responses with identical isotype profiles that were dominated by IgG1 PFC. The role of T cells in B cell differentiation and isotype expression is discussed.
Alteration of in vitro immunoglobulin secretion by amosite asbestos.
Authors: Lawrence E C , McClung H W , Wilson R K , Key M M , Dodson R M , Hurst G A
Published: December 2022

We investigated the in vitro effects of amosite asbestos on immunoglobulin (Ig) secretion by human peripheral blood mononuclear leukocytes (MNL). Concentrations of 100 to 300 micrograms/ml of amosite asbestos reduced the number of Ig-secreting cells recovered from 6-day cultures of unstimulated MNL or MNL stimulated with Epstein Barr virus. By contrast, the Ig secretory response to pokeweed mitogen was enhanced by 10 to 100 micrograms/ml concentrations of amosite asbestos; however, amosite asbestos no longer enhanced the response to pokeweed mitogen when MNL were first partially depleted of monocytes (to less than 2%) esterase-positive cells remaining). These results indicate that amosite asbestos has multiple effects on the cells involved in Ig secretion: 1) amosite asbestos inhibits unstimulated B cell function; 2) amosite asbestos inhibits the function of B cells stimulated with the direct B cell activator Epstein Barr virus; and 3) amosite asbestos may alter regulator monocyte function allowing enhanced Ig secretion in the presence of monocyte-dependent B cell triggers such as pokeweed mitogen.
Selective induction of autoantibody secretion in human bone marrow by Epstein Barr virus.
Authors: Fong S , Vaughan J H , Tsoukas C D , Carson D A
Published: December 2022

The bone marrow is an important site for B lymphocyte differentiation and antibody synthesis in animal and man. However, few experiments have examined directly its immunologic functions in humans. In the present experiments, we have induced bone marrow B lymphocytes from human donors with degenerative arthritis of varying ages to secrete two autoantibodies, IgM and anti-IgG (rheumatoid factor) and IgM anti-human thyroglobulin (Tg), by stimulation with the polyclonal B cell activator Epstein Barr virus (EBV). The EBV-stimulated bone marrow cells secreted significantly more IgM anti-IgG (p less than 0.01) and IgG anti-Tg (p less than 0.01) than matched, identically treated peripheral blood cells. Bone marrow cultures from donors over the age of 60 yr, particularly females, produced more rheumatoid factor than cultures from younger donors (p less than 0.01). The EBV-inducible autoantibodies were immunospecific as demonstrated by adsorption studies. A potential pathogenic role in the inflammatory process was suggested by the finding that the EBV-inducible IgM anti-IgG autoantibodies were capable of activating the classical complement pathway as assessed by the cleavage of C4. These results indicate that the human bone marrow is a selective reservoir for EBV-inducible autoantibody precursor B lymphocytes, and that the size of the reservoir increases with age.
Rapid induction of seven proteins in human lymphocytes by interferon; correlation to natural killer cell activity.
Authors: Gustafsson A , Sundström S , Ny T , Lundgren E
Published: December 2022

The early effects of interferon (IFN) on the synthesis of protein in human nylon wool-nonadherent lymphocytes have been stimulated by use of two-dimensional electrophoresis. IFN-alpha or -beta as well as Escherichia coli-produced IFN-alpha 2 induced the rapid formation of seven proteins (Mr 80, 75, 62, 53, 38, 36, and 33 kD). At least five proteins were expressed within 2 hr of incubation with IFN. The synthesis of the seven proteins seemed to require rapid transcription of new RNA, because actinomycin D markedly inhibited their formation only when added less than 30 min after IFN. A good correlation was found between the ability of actinomycin D to inhibit both the formation of new proteins and the augmentation of natural killer (NK) cell activity. Screening of a panel of 10 hematopoietic and two anchorage-dependent cell lines revealed that p62 and p38 were induced in most cell lines, whereas p80 and p33 were preferentially induced in lymphoid cell lines. Three proteins could not be induced by IFN in any of the 12 cell lines, and thus could represent molecules mediating differentiated functions, possibly involved in NK cell function.
Studies on the contribution of putative anti-carbohydrate antibodies to the specificity of immunofluorescence staining of murine T cells with chicken anti-mouse (Fab')2 reagents.
Authors: Deluca D , Warr G W , Marchalonis J J
Published: December 2022

Despite a large body of immunochemical data indicating that chicken antibodies specifically recognize immunoglobulin-like structures on mammalian T and B cells, the possibility has been raised that the fluorescence staining of murine T cells with chicken antibodies directed against the (Fab')2 fragment of pooled mouse IgG might be due to carbohydrate moieties shared by heavily glycosylated proteins such as fetuin and mucin. Therefore, the presence of antibody activity against these glycoproteins in chicken antisera to mouse IgG (Fab')2 fragments and to MOPC-41 (murine kappa chains) was tested using a double antibody radioimmune assay. No detectable binding of fetuin or mucin was found in our chicken anti(Fab')2 reagent, whereas activity against (Fab')2 fragments was readily detected. In spite of the lack of anti-mucin or anti-fetuin activity, these glycoproteins were capable, along with fetal calf serum, whole mouse IgG, and mouse (Fab')2 fragments, of inhibiting the binding of anti-mouse (Fab')2 to T cells assayed by double immunofluorescence. However, the effective inhibition by fetuin only occurred when very large amounts of the glycoprotein were used, and titration experiments indicated that fetuin, is on a molar basis, at least 60-fold less efficient and inhibitor of the fluorescence reaction than (Fab')2 and nearly 3000-fold less efficient on a molar carbohydrate basis. We conclude that chicken anti-(Fab')2 and anti-mouse kappa chains stain T cells specifically via an immunoglobulin-like determinant and not by anti-carbohydrate activity.
The analysis with monoclonal antibodies of the heterogeneity of Ia glycoproteins on chronic lymphocytic leukemia cells.
Authors: Addis J B , Tisch R , Falk J A , Letarte M
Published: December 2022

The accessible Ia molecules on the surface of chronic lymphocytic leukemia (CLL) cells were quantitated in the cellular radioimmunoassay with saturating concentrations of monoclonal antibodies. Monoclonal antibody 21w4, like DA/2 antibody, recognizes monomorphic determinants of human Ia antigens. The amount of 21w4 or DA/2 bound to CLL cells derived from eight patients (varying from 2.6 to 13.9 x 10(5) molecules/cell) appears to be the maximum observed with the antibodies studied. Two other antibodies, 18d5 and 21r5, although also directed at nonpolymorphic Ia determinants, bind differentially to CLL cells, with the ratios of 21r5/21w4 and 18d5/21w4 varying from 0.08 to 0.90. Sequential immmunoprecipitation studies have established that the four epitopes 18d5, 21r5, 21w4, and DA/2 were present on the same molecules. All Ia molecules express 21w4 and DA/2 epitopes, whereas only certain subsets of Ia molecules carry accessible 21r5 or 18d5 epitopes. Competitive binding studies showed that the epitopes recognized by the four monoclonal antibodies were different. Monoclonal antibodies 21r5 and 21w4 did not inhibit each other's binding. Furthermore, binding of 21w4 to CLL cells potentiated the binding of 125I-21r5 IgG to the same cells, suggesting that binding of 21w4 antibody induces a conformational change in the molecule that renders 21r5 epitopes more accessible.
T cell responses to select Ia determinants using the I-A mutant mouse strain B6.C-H-2bm12.
Authors: Skelly R R , Pappas F , Koprak S , Ahmed A , Hansen T H
Published: December 2022

The T cell repertoire of B6.C-H-2bm12 mice (an I-A mutant mouse strain) to wild-type Iab antigens was investigated using both secondary proliferative cultures and cloned T cell lines. Because bm12 mice have a gain-loss mutation of their gene encoding the Ia beta-chain polypeptide, bm12 anti-B6 T cell responses are specific for the select component of Iab specificities that was lost as a result of the mutation. Although stimulator cells bearing Iab antigens elicited the strongest responses, Iaq, d, and s antigens also resulted in reproducible stimulations of these bm12 anti-B6-primed T cells. Cloned T cell lines isolated from bm12 anti-b6 cultures revealed similar findings, with most clones recognizing determinants unique for Iab antigens; however, clones showing cross-reactions with Iad and/or q were also selected. Using F1 hybrid responder T cells (mutant x cross-reactive strain), we further dissected this cross-reactivity into several distinct cross-reactive determinants. Because bm12 mice lack the serologically defined Ia differentiation antigen W39, T cell recognition of this determinant was investigated by using bm12 anti-B6-primed cells. Stimulation by Ia.W39+ cells was appreciably better than by Ia.W39- (Xid-defective) cells, suggesting that bm12 T cells recognize an Xid-regulated, W39-like Ia differentiation antigen.
Cellular sensitivity to collagen in bleomycin-treated rats.
Authors: Schrier D J , Phan S H , Ward P A
Published: December 2022

Previous reports have suggested that immune mechanisms are involved in the induction of certain types of pulmonary fibrosis. In this study endotracheal bleomycin was used to induce pulmonary fibrosis in rats, and they were then examined for cellular sensitivity to homologous interstitial collagens isolated from lung. Results indicate that rats treated with bleomycin develop transient cellular sensitivity to homologous collagen. Immunity appears to be selective to type I collagen with minimal response to type III collagen. The kinetics of the development of autoimmunity paralleled the transient increase in collagen synthesis in response to bleomycin treatment. This response was abolished upon prior treatment of the challenging antigen with purified bacterial collagenase, but was resistant to trypsin digestion. This finding confirms the true collagenous nature of the stimulating antigen and eliminates the possibility of a noncollagenous contaminant as the effective agent in the antigen preparation. The data suggest that cellular sensitivity to homologous collagen in response to bleomycin treatment may play a role in the overall fibrotic response.
Properties of a new complement-dependent bactericidal factor specific for Ra chemotype salmonella in sera of conventional and germ-free mice.
Authors: Kawakami M , Ihara I , Suzuki A , Harada Y
Published: December 2022

A complement-dependent bactericidal factor, Ra-reactive factor, that acts specifically on Ra chemotype strains of Salmonella was found in sera of eighteen strains of nonimmune mice, including a thymusless strain. The specificity of binding of this factor to the cells of Ra chemotype bacteria was evidenced by absorption studies. The factor was inactivated by treatment with 0.02 M 2-mercaptoethanol or heating for 30 min at 55 degrees C. This factor was present in mice reared by feeding with a chemically defined, antigen-free diet under germ-free conditions. Fetal and newborn mice from a conventional or germ-free mother had the factor, although the titer was lower than the adults'. The titer in the newborn mice reached adult level 1 wk after birth. The titer in the adult mice did not increase markedly after immunization with heat-killed bacteria or administration of Freund's complete adjuvant and turpentine. These results indicate that this factor is actively produced by mice that have never been stimulated with exogenous Ra antigen.
Immunologic deficiency during experimental Chagas' disease (Trypanosoma cruzi infection): role of adherent, nonspecific esterase-positive splenic cells.
Authors: Kierszenbaum F
Published: December 2022

The involvement of adherent splenic cells in the production of deficient lymphocyte responses during the acute phase of experimental Chagas' disease was investigated. When cultured together, purified adherent splenocytes from mice acutely infected with Trypanosoma cruzi caused a significant reduction in the responses of normal mouse spleen cells to T and B cell-specific mitogens. Similar observations were made when infected mouse adherent splenocytes were co-cultured with normal mouse nonadherent cells. Exchange of adherent cells in infected mouse spleen cells suspensions for adherent cells from uninfected mice resulted in increased responses to stimulation with the T and B cell mitogens tested. Treatment of infected mouse cell suspensions with indomethacin improved the responsiveness of these cells to the mitogens. These results support the concept that the immunosuppression that is characteristic of experimental acute Chagas' disease is at least in part mediated by an adherent cell population and is dependent on a prostaglandin-mediated mechanism.
Defective natural cytotoxicity in patients with cancer: normal number of effector cells but decreased recycling capacity in patients with advanced disease.
Authors: Steinhauer E H , Doyle A T , Reed J , Kadish A S
Published: December 2022

Patients with advanced cancer exhibited lower natural cytotoxicity against K562 target cells in a standard short-term chromium release assay than did patients with localized malignancy or normal individuals. Although natural killer (NK) activity of advanced cancer patients could be augmented by nylon column depletion of adherent cells and enriched further by fractionation of discontinuous Percoll gradients, the NK level attained remained below that obtained from PBL of normal donors treated by the same procedures. The pattern of NK activity obtained on the Percoll gradients was the same for all individuals studied. The proportion of large granular lymphocytes (LGL) paralleled NK activity and was highest in the peak NK fraction in all individuals tested. Advanced cancer patients with low NK activity showed no decrease in number of LGL from that of other cancer patients or normals. Similarly, there was no decrease in target binding cell number. Using a single-cell assay in agarose, we found that the number of active NK cells was the same for all patients, whether with localized or advanced malignancy, and normal subjects. In a 3-hr time period, there were no differences in the rate of killing in agarose. The maximum killing potential (Vmax) of the NK-deficient advanced cancer patients in a 51Cr-release assay was significantly lower than that of normals or other cancer patients. The observed defect in natural cytotoxicity of these patients thus appeared to be due to a reduced recycling capacity.
Antibody affinity may influence antigenic modulation of the common acute lymphoblastic leukemia antigen in vitro.
Authors: Lebien T W , Boué D R , Bradley J G , Kersey J H
Published: December 2022

We have produced a new monoclonal antibody designated BA-3. An extensive side-by-side serologic comparison of BA-3 with the anti-common acute lymphoblastic leukemia antigen (CALLA) monoclonal antibody J-5 was undertaken. Cells examined included established leukemic cell lines, malignant cells from patients with newly diagnosed leukemia/lymphoma, and normal hematopoietic tissues. In all experiments the cellular distribution of the antigens recognized by BA-3 and J-5 were identical when analyzed by immunofluorescent microscopy and the FACS. Iodination of NALM-6 cells, followed by radioimmunoprecipitation and SDS-PAGE, indicated that BA-3 (like J-5) precipitated a glycoprotein of approximately 100,000 daltons. Competitive binding studies using 125I-labeled BA-3 indicated that BA-3 and J-5 were binding to closely associated (if not identical) epitopes on CALLA. Calculation of the equilibrium constant (K value) for BA-3 and J-5, and the approximate number of CALLA molecules per cell, was graphically determined using Scatchard plots. BA-3 and J-5 were shown to have K values of approximately 2.7 x 10(7) M-1 and 7.2 x 10(7) M-1, respectively, with NALM-6 cells expressing 1 x 10(5) to 2 x 10(5) CALLA molecules per cell. Additional studies with BA-3 failed to demonstrate significant antigenic modulation of CALLA in vitro using fresh leukemic cells and leukemic cell lines. Thus, we suggest that antibody affinity may be a significant factor influencing antigenic modulation of CALLA in vitro.
Further Observations on the Toxins of Hemolytic Streptococci
Authors: Kirkbride Mary B. , Wheeler Mary W.
Published: December 2022

It is obviously important that a definite conclusion be reached as to whether, with present methods, it is possible to distinguish a specific group of scarlet fever streptococci. Although agglutination reactions are undoubtedly of value in bringing out certain variations, any differentiation of practical significance must be based ultimately on the action of the toxin.

In a report presented last year (1) we recorded the results of studies with a hundred and fifty strains of hemolytic streptococci from different sources, tested on goats. The results of these tests indicate no fundamental qualitative or quantitative differences in the toxins of scarlet and non-scarlet fever streptococci.

Similar observations have since been recorded. Dr. Anna Williams (2), Dr. E. C. Rosenow (3) and other investigators find that hemolytic streptococci from infections other than scarlet fever frequently produce toxins similar to those of the scarlet fever streptococci and they also report variable reactions in different individuals.
The Blood-Groups Among the Laps in Sweden
Authors: Rietz Torsten
Published: December 2022

An investigation of the blood-groups among Swedish Laps shows (1) a proportion of 51 per cent group I, 42 per cent group II, 3 per cent group III and 4 per cent group IV. (2) that the proportion between I, II and III differs from hitherto examined peoples, and (3) that the examination of the blood is not in accord with findings in mongolian races.
A Simple Reaction for Detecting the Binding of Agglutinins by Difficultly Agglutinable Suspensions
Authors: Mudd Stuart
Published: December 2022

A simple reaction is described which has proved satisfactory for detection of the binding of agglutinins by certain difficultly agglutinable bacteria. The reaction is ordinarily positive in the agglutination prezone. It has proved useful in some instances for detecting sensitization of cells which undergo spontaneous sedimentation.
Alcohol Soluble Specific Substances of B. Diphtheriae and of Streptothrix
Authors: Freund J.
Published: December 2022

The antigens of the bacterial suspension and the alcoholic extract of the diphtheria strain used in our experiment could not be differentiated in the absorption experiment. The diphtheria toxin is a different antigen from both the alcohol soluble antigen and the antigens demonstrable in our non-toxic diphtheria bacillus emulsion. The alcohol soluble specific substance of the B. diphtheriae is soluble in methyl alcohol, aceton and weak alkali; insoluble in ether, weak acids and water. The N content of two purified preparations was 5.6 and 5.7 per cent respectively. After hydrolysis the biuret reaction was negative, the ninhydrin reaction was positive. Pepsin and trypsin did not increase the potency of a purified preparation for the complement fixation test. The unit for the complement fixation test in the purified preparation was from 0.00035 to 0.00045 mgm. The addition of an alcoholic solution of lecithin to the alcoholic extract of B. diphtheriae increases its potency in the complement fixation test four times. The solubility of this specific substance makes its protein nature improbable.

An alcohol soluble specific substance was demonstrated in a streptothrix strain. The unit for the complement fixation test in a non-purified preparation was 0.0006 mgm. The addition of an alcoholic solution of lecithin to the alcoholic extract of streptothrix increases its potency in the complement fixation test 200 times in a mixture containing 2 per cent alcoholic extract.

No alcohol soluble specific substance could be demonstrated in the case of B. xerosis, of trychophyton, actinomyces and yeast.
Studies in Specific Hypersensitiveness
Authors: Spain W. C. , Cooke Robert A.
Published: December 2022

With the use of a 95 per cent ethyl alcoholic extract of Toxicodendron radicans, (L.), (Rhus radicans), it is possible to reproduce, in susceptible individuals, the vesicular lesion typical of “poison ivy” dermatitis. It is also possible to confer varying degrees of protection to such susceptible individuals by means of hypodermic injections of the same alcoholic extract. A description of this mode of treatment, with its results, is given here, together with a discussion of later refinements in the preparation of the extracts and in the technic of treatment.

The extract, as first employed by us, was prepared in the following manner: fresh, whole leaves of Toxicodendron radicans, with all excess moisture removed, were placed in clean glass jars, and were covered with 95 per cent ethyl alcohol, in a proportion of 1 gram of leaf to 3 cc. of alcohol. The jars were then tightly capped, and placed in the ice box.