bluuebunny/crossref_metadata_2025 huggingface dataset:
Antibody Responses of Tumor-Bearing Mice to Their Own Tumors Captured and Perpetuated As Hybridomas
Authors: Simrell Charles R. , Klein Paul A.
Published: December 2022

Experiments were performed to determine whether antibody-producing hybridomas could be derived by fusion of BALB/c P3X63Ag8 myeloma cells and spleen cells from C57BL/6 mice either immune to or bearing a syngeneic methylcholanthrene (MCA)-induced fibrosarcoma (M3). Mice were immunized by tumor amputation followed by injections of tumor cells, purified tumor cell membranes, or murine leukemia virus isolated from M3 cells (M3-MuLV). Tumor-bearer spleens were obtained at weekly intervals over a 6-week course of tumor bearing. Hybridoma cultures showing the best growth in selective HAT medium were screened by radioimmunoassay for antibody production against a) viable and glutaraldehyde-fixed cultured M3 cells, b) glutaraldehyde-fixed in vivo-passaged M3 cells, c) purified M3 cell membranes, and d) M3-MuLV. Stable antibody-producing cloned hybridomas were obtained only from fusions involving tumor-bearer spleen cells. One clone (2C2-A1) secretes an IgM antibody that binds well to purified M3-MuLV and M3 membranes, but only weakly to intact M3 cells. Another cloned line (3A5-L1) secretes an IgG1 antibody that binds to M3 cells but not to M3-MuLV. Further analysis of specificity by binding and absorption studies involving more than 30 other normal and transformed murine cell types indicates that 3A5-L1 antibody binds to some, but not all, MCA-induced murine fibrosarcomas. 3A5-L1 antibody demonstrates little, if any, binding to murine tumors of other histologic types or to normal tissues. The results demonstrate that it is possible to capture and perpetuate part of the tumor-bearing host's antibody repertoire in the form of hybridomas secreting monoclonal antibodies specific for tumor-associated antigens.
Mechanism of Action of Human B Cell-Activating Factor
Authors: Wood David D.
Published: December 2022

Human monocytes have been reported to release factors that can elicit distinct responses from a number of different target cells. Three of these activities are compared in this report. Supernatants of endotoxin-stimulated monocytes have been shown to stimulate the direct plaque response of T cell deficient splenocytes in vitro (BAF assay), to stimulate the thymidine incorporation by thymocytes (mitogen assay), and to synergize with phytohemagglutinin in the stimulation of thymocytes (PHA-enhancing assay). By gel filtration, plaque stimulating activity was recovered as a single peak with a m.w. of approximately 15,000 daltons when tested on nude splenocytes. When assayed on normal C57BL/10CR splenocytes, a second peak of plaque-stimulating activity was observed near 35,000 daltons. Mitogenic activity was observed both within the albumin peak and coincident with the 15,000 dalton plaque stimulator. Finally, PHA-enhancing activity was found to be broadly distributed between 8000 and 66,000 daltons, and included molecules that co-eluted with the 35,000 and 15,000 dalton plaque stimulators. Upon further purification of the 15,000 dalton material by isoelectric focusing, the plaque-enhancing, mitogenic, and PHA-enhancing activities co-eluted. This observation raised the possibility that all three activities of the 15,000 dalton material were due to the effects of a single factor on T cells. Some investigators favor this possibility based upon the study of the supernatants of human mixed leukocyte cultures (XRF). Here it is shown that spleen cell populations can be prepared that responded to BAF but not XRF, and vice versa. Therefore, studies on XRF cannot be used as evidence for the mechanism of the action of BAF. These results raise the possibility that this single factor may effect more than one target cell and may be responsible for a number of the sequellae of the stimulation of the reticuloendothelial system by bacterial products.
Purification and Characterization of a Single Chain Precursor C3-Protein (PRO-C3) from Normal Human Plasma
Authors: Minta Joe O. , Ngan Bo-Yee , Pang Anthony S. D.
Published: December 2022

Fresh human plasma was treated with proteinase inhibitors and passed through an immunoadsorbent column of Sepharose anti-C3 globulin. The insolubilized C3 was eluted with 5 M guanidine and, after extensive dialysis, was reduced with 0.2 M 2-mercaptoethanol and electrophoresed on SDS-polyacrylamide gels. The bulk of the eluted C3 dissociated into two major protein bands, m.w. 125,000 and 75,000 corresponding to the α-and β-chains of C3. In addition, a nonreducible single polypeptide chain (SPC) with a m.w. value of 197,000 ± 2,000 similar to the apparent m.w. of unreduced C3 was consistently present. SPC has been purified by elution from SDS (SPC*) and found to remain a single polypeptide chain upon re-electrophoresis on SDS gels in the presence of 0.2 M 2-Mercaptoethanol. The purified SPC reacted with antisera to denatured C3, C3α, and C3β chains. Additionally, antisera to SPC*, also reacted with denatured C3, C3α-, and C3β-chains, revealed a reaction of identity between SPC and C3, and detected partial identity between SPC and C3α- as well as C3β-chains. This suggested that SPC and C3 are antigenically related. The amino acid compositions and tryptic peptide maps of SPC and C3 were similar. Based on these findings, it is suggested that SPC must represent a single-chain precursor C3 (pro-C3) in plasma that escaped post-synthetic proteolytic cleavage into a two-subunit chain C3 molecule.
Synthesis of a Lipopolysaccharide Designed to Conjugate Haptens and Proteins to Erythrocytes for Use as Target Cells in Passive Hemagglutination and Hemolytic Assays
Authors: Bankert Richard B. , Mayers George L. , Pressman David
Published: December 2022

The synthesis of a reagent that is capable of conjugating a wide variety of haptens and proteins to cell membranes is presented. A very large chemically reactive glucose polymer (i.e., oxidized dextran), to which fatty acid side chains of myristic acid have been added, covalently binds to amino groups of haptens or proteins forming a Schiff base. The protein or hapten myristoyl dextran compounds attach directly to cell membranes when incubated with erythrocytes. The attachment of the ligands to the cell presumably occurs via the intercalation of the lipid portion of the myristoyl dextran compounds into the hydrophobic lipid bilayer of the plasma membrane. The conjugation of haptens and proteins to plasma membranes via this synthetic lipopolysaccharide reagent has several significant advantages over other coupling procedures: 1) conjugation of ligands occurs rapidly at conditions that are physiologic with respect to the cell, 2) both haptens and proteins can be conjugated via the same procedure, 3) the conjugating reagent, i.e., hapten-myristoyl dextran or protein-myristoyl dextran, is chemically stable and can be stored for extended periods of time at 4°C, 4) target cells prepared via myristoyl dextran detect picogram levels of antibody in hemolytic assays without any detectable nonspecific lysis, and 5) a very small amount of the hapten or protein myristoyl dextran reagent is required to produce target cells (i.e., as little as 1 to 10 µg of hapten).

The following compounds have been successfully conjugated to cell membranes by using myristoyl dextrans as the linking reagent; N-ε-DNP-lysine, 3-aminopyridine, 4-aminopyridine, 4-aminobenzenearsonate, 4-aminobenzoate, 4-aminophthalate, 5-aminoisophthalate, Bence Jones protein, human γ-globulin, bovine γ-globulin, rabbit anti-human Fab antibody, and mouse anti-phthalate antibody. The effective use of myristoyl dextran-conjugated erythrocytes as target cells in the passive hemagglutination assay, the hemolytic spot test, the Jerne plaque assay, and an antigen-mediated hemolytic assay is established and the sensitivity of the target cells is quantitatively compared in these assays to target cells prepared by other commonly used procedures.
Induction of Lysosomal Enzyme Secretion by Alveolar Macrophages in Response to the Purified Complement Fragments C5a and C5a des-arg
Authors: McCarthy Kathleen , Henson Peter M.
Published: December 2022

Purified C5a and its “inactive” form, C5a des-arg, were shown to induce secretion of acid hydrolases from rabbit alveolar macrophages (AM) in a concentration-dependent manner. Secretion increased with time to 5 times above controls by 72 hr. Concentrations of these enzymes in the cell lysates did not decrease during the incubation, suggesting that synthesis of new enzyme was occurring.

The lysosomal enzyme secretion was accompanied by increased pinocytosis and release of proteolytic enzymes from the macrophages. At no time was significant lactic dehydrogenase liberated, indicating that secretion was selective and not due to cell death.

Data presented also suggest that C5a des-arg induced secretion from the macrophages of a chemotactic factor for neutrophils. It was concluded that C5a and C5a desarg may play a role in lung injury by interactions with AM, inducing the secretion of acid hydrolases and proteolytic enzymes that can cause tissue damage, and by regulating the influx of other inflammatory cells into the interstitium and air spaces.
Macrophages in Resistance to Rickettsial Infection: Macrophage Activation in Vitro for Killing of Rickettsia Tsutsugamushi
Authors: Nacy Carol A. , Meltzer Monte S.
Published: December 2022

Rickettsia tsutsugamushi, strain Gilliam, replicates in cultures of resident peritoneal macrophages from BALB/c mice. Macrophage cultures treated with culture supernatants of spleen cells from rickettsial-infected mice stimulated with heat-killed rickettsiae markedly suppressed macrophage infection by rickettsiae. Rickettsiacidal activity of activated macrophages was dependent upon both lymphokine concentration and time of incubation in lymphokines. Treatment of macrophage cultures with lymphokines before exposure to viable rickettsiae resulted in an immediate decrease in percent macrophages infected and numbers of viable intracellular rickettsiae. In these cultures, enhanced intracellular killing was also apparent with further incubation (24 hr). The immediate effect of lymphokine-pretreated macrophages was dissociated from intracellular killing by infecting macrophage cultures first and adding lymphokines after infection. In these cultures, both percent macrophages infected and titers of viable intracellular rickettsiae were dramatically reduced as well.
Sex-Related Immunocompetence of BALB/c Mice
Authors: Krzych Urszula , Thurman Gary B. , Goldstein Allan L. , Bressler Joseph P. , Strausser Helen R.
Published: December 2022

The responses of lymphoid cells from the thymus, lymph nodes, and spleen of male and female BALB/c mice were evaluated to determine if sex-related variations in immune expression could be found. Immunologic assays used included blastogenic responses to mitogens, mixed lymphocyte responses, and direct and indirect measurement of plaque-forming cells against soluble and particulate antigens. The results indicated that responses of spleen cells from young adult female mice were higher than those of males in all comparative tests. Little or no differences between the sexes were observed in the mitogenesis of lymph nodes and thymuses. Newborn mice did not demonstrate the sex-associated immune differences. Among the weanling mice slight differences between male and female spleen cells responsiveness to mitogenic agents were observed.
Prevention of Transplantable Tumors by Adoptive Transfer of Spleen Cells from Immunized Rats
Authors: Fish Donald C. , Djurickovic Draginja B. , Huebner Robert J.
Published: December 2022

Fischer 344 rats were specifically hyperimmunized with allogeneic, nonvirus-producing [Kirsten murine sarcoma virus (KiMSV)] or syngeneic, virus-producing [KiMSV (Rasheed)] rat tumors. Spleen cells taken from these rats adoptively transferred protection against a 100 to 1,000 X rat tumor dose50 cell challenge with several different transplantable rat tumors. Protection was obtained with spleen cells after removal of adherent cells and macrophages but not peritoneal cells. The spleen cells were not directly cytotoxic but required more than 3 days residence in the recipient before protecting the recipient against challenge. No protection against tumor cell challenge was observed when spleen cells were lethally x-ray irradiated before injection into nontreated rats. Spleen cells taken from rats immunized with normal histocompatibility antigens did not protect in this test system.
Imbalances of T Cell Subpopulations in Patients with Atopic Diseases and Effect of Specific Immunotherapy
Authors: Canonica Giorgio Walter , Mingari Maria Cristina , Melioli Giovanni , Colombatti Marco , Moretta Lorenzo
Published: December 2022

Reduced numbers of T cells with Fc receptors for IgG (TG cells) are present in blood samples of patients with respiratory allergic disease, mainly those with severe symptoms, TG cells have been previously shown to be suppressor in the pokeweed mitogen- (PWM) dependent B cell differentiation. T cells with Fc receptor for IgM (TM cells), which help immunoglobulin production, are in a normal range. After specific hyposensitization, resulting in a sharp improvement of clinical symptoms, TG cell subset reached normal values.
Ligand-Activated T Cell Growth Factor-Induced Proliferation: Absorption of T Cell Growth Factor by Activated T Cells
Authors: Bonnard G. D. , Yasaka K. , Jacobson D.
Published: December 2022

The fate in culture of the T cell growth factor (TCGF), which is required for continued growth of human cultured T cells (CTC) in vitro, was studied. TCGF activity was stable for 7 days at 37°C. However, it was no longer detectable after incubation with actively growing CTC at 37°C for 3 days. This loss of TCGF activity also occurred quite rapidly and was detectable within 1 hr of incubation of 0.3 ml supernatant with 2 to 5 × 107 CTC at 23°C. 2 × 108 mononuclear peripheral blood leukocytes were not effective in removing TCGF activity, and incubation with similar numbers of cells from B and T cell lines had no effect. Three-day-old concanavalin A and phytohemagglutinin blasts were very reactive with TCGF, so that 107 or 2 × 107 cells consistently removed TCGF activity. These experiments suggested specific absorption of TCGF by activated T cells, and led us to develop a model of ligand-activated TCGF-induced proliferation of T cells: Ligands induce production of TCGF by T-producer cells and deliver a first signal to the T-responder cells. This causes a receptor for TCGF to appear on T-responder cells. Only then does TCGF deliver the obligatory second signal that is needed to drive the T-responder cells into proliferation.
Regulation of the Murine IgE Antibody Response
Authors: Schwenk Robert J. , Kudo Koichiro , Miyamoto Motoko , Sehon Alec H.
Published: December 2022

Anti-ovalbumin (OA) IgE antibody responses were measured in B6D2F1 mice as a function of time and antigen dose. One hundred to 200 µg of OA in Al(OH)3 elicited transient responses, whereas 1 to 10 µg of OA in Al(OH)3 elicited persistent anti-OA IgE responses of high titer. T cells isolated from the spleens of mice mounting either a persistent or a transient response strongly suppressed primary anti-DNP IgE responses in unirradiated recipient mice that were immunized with DNP-OA in Al(OH)3; it was, therefore, concluded that suppressor T cells (Ts cells) were activated during both the persistent and transient IgE responses. Nevertheless, in the present study it was not possible to completely rule out the contention that IgG antibodies may also have been suppressing the IgE response. With a modified adoptive transfer system, it was shown that these Ts cells were sensitive to low doses (250 R) of x-irradiation. The suppressive activity of long-term OA primed cells was also shown to be markedly enhanced when cultured for 24 hr with soluble OA; this finding was interpreted to indicate the presence of memory suppressor cells.
Defective Fc Receptor-Mediated Phagocytosis in C3H/HeJ Macrophages
Authors: Vogel Stefanie N. , Rosenstreich David L.
Published: December 2022

Macrophages derived from LPS-unresponsive C3H/HeJ mice lose their ability to phagocytose opsonized SRBC (EA) after 24 hr of in vitro culture, whereas macrophages from normal mice exhibit a marked increase in phagocytosis of EA under the same culture conditions. Defective phagocytosis is not due to a decrease in macrophage number or viability and appears to be specific for Fc receptor-mediated phagocytosis, since phagocytosis of latex particles by C3H/HeJ macrophages is normal. The phagocytic defect appears to be due to a decrease in the binding ability or number of Fc receptors, since C3H/HeJ macrophages exhibit a loss of Fc binding ability that parallels the loss in their phagocytic ability. Cultured macrophages from LPS responsive (C3H/HeN) mice maintain their ability to bind EA after 48 hr culture.

This Fc receptor defect appears to be secondary to a failure of C3H/HeJ macrophages to maintain a differentiated state in vitro, since induction of differentiation of C3H/HeJ macrophages with the supernatant derived from Con A-treated spleen cells (CS) results in an almost complete restoration of both their ability to bind and phagocytose EA. The activity of CS appears to be due to a lymphokine, since it is not present in the culture supernatants of unstimulated spleen cells and is not produced by Con A alone.

These findings emphasize the close association between the induction of macrophage differentiation and the sensitivity of these cells to LPS. Furthermore, they suggest that one function of the Lps gene may be to regulate the ability of cells to respond to physiologic differentiation signals in addition to or instead of lipid A.
Further Characterization of the Human Inducer T Cell Subset Defined by Monoclonal Antibody
Authors: Reinherz Ellis L. , Kung Patrick C. , Goldstein Gideon , Schlossman Stuart F.
Published: December 2022

Evidence is presented that the OKT4+ T cell subset in man, defined by a monoclonal hybridoma antibody, provides help for B lymphocyte differentiation in a PWM driven system. Both B cell proliferation and intracytoplasmic immunoglobulin synthesis are facilitated by OKT4+ and not by OKT4- T cells. Given earlier studies demonstrating that OKT4+ T cells were necessary for generation of T cytotoxic cells and the present study that OKT4+ T cells are necessary for the differentiation of B cells, it would appear that the OKT4+ population is the major human T helper (inducer) subset.
The Qa-2 Antigen on Lymphocyte Subpopulations. Mixed Lymphocyte Culture and Cell-Mediated Lympholysis
Authors: Sullivan Karen A. , Flaherty Lorraine
Published: December 2022

Treatments of spleen cells from Qa-2+ strains with Qa-2 antiserum plus complement (C) have revealed that the Qa-2 antigen is present on restricted functional lymphocyte subpopulations. Anti-Qa-2 plus C reduced the mixed lymphocyte culture response and inhibited the generation of cytolytic effector cells. This treatment, however, did not affect cytolytic effector cells once they were generated.
A new lymphocyte surface protein present in normal urine. II. Cellular distribution and biologic properties.
Authors: Velosa J A , Calvanico N J , Anderson W L , Tomasi T B
Published: December 2022

A protein component present in normal human urine has been found on the surface of epidermal cells and lymphocytes. This protein, called urinary acidic antigen (UA), can not be detected in concentrated fractions of normal human serum by double immunodiffusion, suggesting that it is quickly cleared from the circulation. It is readily detected, however, in sera of patients with renal failure. Although it can be eliminated from the cell surface by repeated washings with PBS, it was shown to cap with anti-UA-specific antiserum. Anti-UA suppresses PWM-induced proliferation, but not the lymphocyte response to PHA, Con A, or allogeneic cells. Thus UA appears to have a specific relationship to the pokeweed response. Whether it is a structural component of the PWM receptor is uncertain.
Regulation of myeloma growth in vitro by idiotype-specific T lymphocytes.
Authors: Flood P M , Philipps C , Taupier M A , Schreiber H
Published: December 2022

Spleen cells of BALB/c mice immunized with purified BALB/c myeloma protein (MP) caused idiotype-specific suppression of DNA synthesis and colony formation by myeloma cells in vitro. Lymphocytes from mice immunized with 315-MP inhibited only MOPC-315 plasmacytoma cells; conversely, lymphocytes from mice immunized with 167-MP inhibited only MOPC-167 plasmacytoma cells. Antisera from those mice that showed cell-mediated inhibition of myeloma growth had no significant effect. The suppressive activity of the spleen cells was markedly reduced by treatment with anti-Thy 1.2 and complement, and the immune cells responsible for the suppression did not adhere to nylon wool. The suppressor cells adsorbed to myeloma protein-coated plates, and after elution, inhibited myeloma cells specifically at a 1:1 effector to target cell ratio. Suppression of myeloma growth was due to cytostatic rather than cytolytic effects. These findings suggest that mice immunized with purified myeloma protein have idiotype-specific T cells that can regulate myeloma cell growth. This mechanism may provide an explanation for the transplantation resistance to myelomas induced by immunization with myeloma protein. Similar idiotype-specific T cells may also play a role in the idiotype-specific regulation of immune responses by acting late in differentiation to block the proliferation of antibody-secreting normal B cells.
Analysis of cytotoxic effector cell function in patients with leukemia or aplastic anemia before and after marrow transplantation.
Authors: Livnat S , Seigneuret M , Storb R , Prentice R L
Published: December 2022

Cytotoxic effector cells mediating natural killing (NK), antibody-dependent cellular cytotoxicity (ADCC), and lectin-dependent cellular cytotoxicity (LDCC) were studied in patients with leukemia or aplastic anemia before and/or after marrow transplantation. Before transplantation, about one-third to one-half of the patients were deficient in cytotoxic activity. In patients with leukemia, this was most likely due to large numbers of circulating blast cells diluting or replacing the effector cells. In patients with aplastic anemia there was an apparent absence of the effector cells in a proportion of the patients. After marrow transplantation, cytotoxic activity in all three systems returned to normal rapidly, by 30 days, and remained so through 100 days. However, about 20% of patients studied beyond 1 year were deficient in these functions. There were no significant associations between cytotoxic activity and important clinical parameters including infections, graft-vs-host disease, and recurrence of leukemia. Our findings do not support an immunosurveillance role for NK against leukemia after marrow transplantation. Furthermore, they point out the need for new in vitro approaches for meaningful monitoring of marrow transplant patients. Finally, our results showed a significant discordance between NK, ADCC, and LDCC activities in these immunologically perturbed individuals, indicating that either different cell populations or different cellular mechanisms are involved in these cytotoxic functions.
Inhibition of the immune response by Fc receptor-bound specific antibody on T lymphocytes.
Authors: Setcavage T M , Kim Y B
Published: December 2022

Removal of naturally occurring membrane-bound antibody from the surface of nonadherent spleen cells by pronase enhances the in vitro plaque-forming cell response to sheep erythrocytes, and this enhancement can be abrogated by preincubation of the enzyme-treated cells in anti-sheep red blood cell antibody. This inhibitory effect is specific in that prior absorption with sheep erythrocytes but not with noncross-reacting red blood cells removes the activity from antibody-containing preparations. Whole IgG molecules are necessary to bind to Fc receptor; F(ab')2 fragments do not inhibit. The functionally significant Fc receptor-bound antibody in this system is found on the T lymphocyte.
Lipopolysaccharide-induced immunomodulation of the generation of cell-mediated cytotoxicity. I. Suppression of the development of cytotoxic lymphocytes.
Authors: Vallera D A , Pflugfelder U , Schmidtke J R
Published: December 2022

Bacterial lipopolysaccharide from a variety of Gram-negative organisms suppresses the development of cytotoxic killer cells in the murine MLC. Cytotoxic T lymphocytes were generated in vitro by incubating BALB/c responder spleen cells with irradiated C57BL/6 stimulator cells for 5 days in mixed lymphocyte culture (MLC). The addition of LPS at the initiation of MLC suppressed killing of 51Cr-labeled target cells in a dose-dependent manner. LPS was active only during the afferent phase of CMC, since it did not interfere with the efferent phase of the assay. Furthermore, timed addition and timed removal studies suggested that the presence of LPS during the first 48 hr of MLC was critical for maximal suppression of CMC. Lipid A extracted from LPS, which had been shown to be highly suppressive when added to the sensitization phase of the CMC assay, was also inhibitory. Moreover, when LPS was added to MLC in the presence of tritiated thymidine, the proliferative activity of the responder cells increased markedly after 72 hr of culture. These data suggest that LPS, a known B cell mitogen, can modulate the complex sequence of cellular interactions that leads to the generation of cell-mediated cytotoxicity.
Macrophage-mediated suppression. I. Evidence for participation of both hdyrogen peroxide and prostaglandins in suppression of murine lymphocyte proliferation.
Authors: Metzger Z , Hoffeld J T , Oppenheim J J
Published: December 2022

Macrophage-mediated suppression of Con A induced proliferation of murine splenic lymphocytes was studied in vitro. Either Corynebacterium parvrum-induced peritoneal exudate cells (PEC) or thioglycollate-induced PEC could totally suppress lymphocyte proliferation at a PEC:lymphocyte ratio of 2:10, whereas a ratio of 1 to 1.5: 10 caused a partial (60 to 68%) suppression. Exogenous PGE1 and PGE2 at concentrations of 10(-8) to 10(-6) M could not totally suppress lymphocyte proliferation. Conversely, indomethacin reversed the partial suppression by macrophages but only partially protected the totally suppressed lymphocyte cultures. Macrophage-mediated cytotoxicity and cytostasis have been proposed to be mediated by hydrogen peroxide. Therefore, hydrogen peroxide was investigated as a possible additional cause for macrophage-mediated suppression, by testing the anti-inhibitory effects of catalase. Partially suppressed cultures were effectively protected from suppression by catalase. In totally suppressed cultures, catalase alone was only minimally effective, but a synergistic effect of catalase and indomethacin was obtained, which provided complete protection from maximal macrophage-mediated suppression. Catalase presumably contributes to the reversal of macrophage suppressive effects both by reducing the direct toxic effect of H2O2 and by preventing the H2O2 from generating additional prostaglandins.
Specific clonal regulation in the response to phosphocholine. I. Genetic analysis of the response of a distinct idiotype (M511 Id).
Authors: Ruppert V J , Williams K , Claflin J L
Published: December 2022

The genetic basis of the BALB/c anti-PC response pattern was studied in allotype and H-2 congenic mouse strains. In both a T-dependent (PC-KLH) and a T-independent (R36A) system, one of the major anti-PC idiotypes, the M511 IdI, is under specific clonal regulation. The expression of the two other major anti-PC idiotypes, T15 and M603X, is independent of such control. The expression of the M511 IdI is controlled by more than one gene, with at least one of the genes unlinked to either the Igh or the H-2 locus. Low responder mice, such as BALB/c, possesses the relevant genetic information coding for the H and L chains of the M511 Idi-positive antibodies, indicating that the expression of this idiotype is dependent on a specific regulatory mechanism.
Nonspecific inhibition of encephalomyocarditis virus replication by a type II interferon released from unstimulated cells of Mycobacterium tuberculosis-sensitized mice.
Authors: Pusateri A M , Ewalt L C , Lodmell D L
Published: December 2022

Unstimulated peritoneal cells (PC) from mice sensitized with nonviable Mycobacterium tuberculosis in an oil-droplet emulsion inhibit encephalomyocarditis virus (EMCV) replication in mouse embryo fibroblast monolayers. Concentrations of mycobacteria ranging from 50 to 500 microgram elicit PC that inhibit EMCV replication greater than 99%. PC collected 2 to 6 weeks post-inoculation of mycobacteria are most effective (greater than 99% inhibition), although cells harvested from mice 7 through 10 weeks inhibit viral replication greater than 90%. Inhibition of replication is not detected unless PC are in contact with infected monolayers for a minimum of 8 to 10 hr; nonviable PC are not effective. Optimal inhibition occurs in cultures infected with a low multiplicity of EMCV that are incubated at 37 degrees C. Inhibition of replication is not due to pH changes or depletion of nutrients in cultures, adsorption and/or inactivation of EMCV by macrophages, or killing of monolayers by the PC. Inhibition of viral replication by the unstimulated PC appears to be due to an interferon that is similar but not identical to classical mouse Type II interferon.
The ninth component of human complement: purification and physicochemical characterization.
Authors: Biesecker G , Müller-Eberhard H J
Published: December 2022

A procedure for the isolation of the human complement (C) protein C9 is described. The procedure allowin. The purified protein has the electrophoretic mobility of an alpha-globulin, and is a single polypeptide chain with a m.w. of 71,000. No impurities were detected either on gel electrophoretic or immunochemical examination. C9 is a glycoprotein containing 7.8% carbohydrate, and in terms of residues per mole, 3.0 glucosamine, 17.6 neutral hexose, and 7.4 sialic acid. Its amino acid composition is typical of a globular serum protein. Upon automated Edman degradation of reduced and alkylated C9, no amino acid residues were released, suggesting a blocked N-terminus. The concentration of C9 in normal human serum is 58 +/- 8 microgram/ml. A high titer rabbit antiserum was produced and employed to immunochemically deplete serum of C9. The CH50 of the C9-depleted serum was identical to that of whole human serum; however, membrane fragments of erythrocytes lysed by C9-depleted serum lacked the typical ultrastructural C lesions, which constitute the dimeric membrane attack complex.
Binding of eosinophil peroxidase to mast cell granules with retention of peroxidatic activity.
Authors: Henderson W R , Jong E C , Klebanoff S J
Published: December 2022

Eosinophil peroxidase (EPO) and to a lesser degree neutrophil peroxidase (myeloperoxidase, MPO) bound tightly to mast cell granules (MCG), particularly when the latter were depleted of histamine by suspension in physiologic salt solutions. The bound EPO was localized on the surface of the granule, and its dissociation required salt concentrations of high enough ionic strength (greater than 0.75 M) to solubilize the MCG matrix. Elution of MPO from the complex occurred at a lower salt concentration. The MCG/EPO complex retained the capacity of the isolated EPO to catalyze the iodination reaction when supplemented with iodide, H2O2, and a protein acceptor and to kill microorganisms when supplemented with H2O2 and a halide (iodide, chloride). Indeed, the MCG/EPO complex had significantly greater iodinating and bactericidal activity than the free enzyme when standardized to equal guaiacol units of peroxidase activity. Thus, in areas of inflammation where mast cells and phagocytic leukocytes coexist, there is the potential for the formation of active complexes extracellularly between mast cell granules and molecules such as EPO (or MPO) that can affect the inflammatory response.
Regulation of macrophage populations. I. Preferential induction of Ia-rich peritoneal exudates by immunologic stimuli.
Authors: Beller D I , Kiely J M , Unanue E R
Published: December 2022

The amounts of Ia-positive and -negative macrophages were studied in peritoneal exudates of normal mice or of mice injected with various inflammatory materials, infected with Listeria monocytogenes, or injected with hemocyanin. Ia-negative macrophages predominated in exudates from normal mice or from mice given mineral oil, peptone, thioglycollate, culture media, or endotoxin. Infection with Listeria caused a very marked increase in Ia-positive macrophages. The induction of Ia-positive macrophages by Listeria inoculation resulted in great part from an immune process. The Ia-positive exudates were more readily generated in immune mice given a secondary challenge with heat-killed organisms. Furthermore, immune T cells transplanted together with heat-killed organisms into normal mice resulted in Ia-rich exudates. Injection of hemocyanin also induced Ia-rich exudates involving an immune process. We conclude that an immune reaction involving T cells regulates the Ia phenotype of the exudate macrophage population. The Ia-positive macrophages were Fc and C3 receptor positive and phagocytized latex particles.
TNP-coupled membranes stimulate T cell proliferation via the macrophage.
Authors: Heber-Katz E , Shevach E M
Published: December 2022

TNP-coupled membranes derived from guinea pig m phi were capable of inducing both primary and secondary antigen-specific T cell proliferative responses; however, effective stimulation by hapten-coupled membranes required the presence of unmodified m phi. A genetic analysis demonstrated that the Ia antigens of the unmodified m phi rather than the Ia antigens borne on the TNP-coupled membranes determined the histocompatibility restrictions on m phi T cell interaction.
A 49,000-dalton polypeptide bearing all antigenic determinants and full immunogenicity of 22-nm hepatitis B surface antigen particles.
Authors: Mishiro S , Imai M , Takahashi K , Machida A , Gotanda T , Miyakawa Y , Mayumi M
Published: December 2022

Spherical 22-nm hepatitis B surface antigen (HBsAg) particles with a subtype adr were purified from plasma of asymptomatic carriers of hepatitis B virus. When purified HBsAg preparation was treated with sodium dodecylsulfate in the absence of reducing agents, it yielded spherical particles with a diameter smaller than 22 nm, and in addition, a polypeptide with a molecular size of 49,000 daltons, which seemed to constitute the outer coat of HBsAg particles. The recovery of the polypeptide on the basis of optical density at 280 nm was 2%, starting from 22-nm HBsAg particles. The 49,000-dalton polypeptide apparently represented a structural unit of the surface of HBsAg particles, since it bore all common (a, Re) and subtypic (d, r) determinants with essentially the same antigenic titers as intact HBsAg particles. Furthermore, this polypeptide was equally immunogenic as 22-nm HBsAg particles in raising corresponding antibodies in mice. When the 49,000-dalton polypeptide was reduced in the presence of 2-mercaptoethanol, it cleaved into 22,000- and 27,000-dalton polypeptides with a drastic decrease in both antigenicity and immunogenicity. These results indicate the different molecular arrangements between outer coat and inner portion of HBsAg particles, and a potential application of the 49,000-dalton polypeptide as a component vaccine, owing to its strong antigenicity both in vitro and in vivo.
Murine cell surface immunoglobulin: two native IgD structures.
Authors: Pollock R R , Mescher M F
Published: December 2022

Murine spleen cell surface IgD is found in two forms upon SDS-polyacrylamide gel electrophoresis under nonreducing conditions. Both IgDI and IgDII are present on the surface of intact cells and appear to be native structures. Neither form could be accounted for by proteolytic degradation or disulfide bond rearrangement. IgDII has an apparent m.w. of 96,000, suggesting an HL structure. IgDI has an apparent m.w. of 150,000, approximately 33,000 daltons lower than that expected for an H2L2 structure. Significant variation in the relative amounts of IgDI and IgDII was found when mice of different strains were examined.
Differential natural killer cell reactivity against T cell lymphomas by cells from normal or stimulated mice.
Authors: Tai A , Burton R C , Warner N L
Published: December 2022

One of the major problems confronting the study of NK cell populations concerns their potential recognition patterns and specificity. In this study three T cell lymphomas that differ markedly in their susceptibility to NK-mediated lysis from normal mice are shown to bear quantitatively but not qualitatively different degrees of NK target structures. The ability of these tumors to be lysed by NK cells can be drastically changed by augmentation of NK activity by LCM virus, but in a manner that differs relatively for the different targets. Comparison of the growth of these tumors in vivo strongly suggests an in vivo action for NK cells.
Cellular requirements for antigen presentation in the induction of a thymus-independent antibody response in vitro.
Authors: Kirkland T N , Sieckmann D G , Longo D L , Mosier D E
Published: December 2022

The ability of various cell populations to bind and present the thymus-independent antigen TNP-Ficoll to a responding cell population was assessed. The in vitro antibody response to TNP-Ficoll depends upon the presence of B lymphocytes and plastic-adherent accessory cells, but does not require T lymphocytes. Purified B cells were the most effective population in binding and presenting TNP-Ficoll, and adherent cells did not perform this function. Antigen binding and presentation was antigen specific and could be blocked with anti-mu antibody, but not by antibodies directed against other immunoglobulin classes. Spleen cells from mice genetically unresponsive to TNP-Ficoll (CBA/N X BALB/c F1 males) were equally effective as normal spleen cells in antigen binding and presentation. We conclude that the initial events in the induction of the antibody response involves antigen binding by B cells, and that subsequent activation of the subset of B cells that can respond to TNP-Ficoll proceeds either via B cell-B cell interaction or B cell-dependent transfer of antigen to macrophage-like cells.
Structural studies on the murine fourth component of complement (C4). IV. Demonstration that C4 and Slp are encoded by separate loci.
Authors: Carroll M C , Passmore H C , Capra J D
Published: December 2022

The S region of the murine major histocompatibility complex (MHC) encodes the structural gene for the fourth complement component (C4) and controls C4 serum levels and expression of the sex-limited protein (Slp). Although Slp was first throught to be an allele of C4, more recent studies have suggested that Slp is either a modified product of the C4 locus or is encoded at a separate locus. By using a sensitive peptide analysis technique, each of the three constitutive polypeptide chains of C4 and Slp was compared, and multiple distinct peptides as well as several shared peptides were detected. These results demonstrate that although C4 and Slp undoubtedly share a common genetic origin, they are encoded by separate structural genes.
Ontogeny of T cell function. I. Acquisition of helper cell activity by the thymus.
Authors: Haines K A , Siskind G W
Published: December 2022

The ontogeny of the ability of thymus cells to "help" the response to a T-dependent antigen was studied in a cell transfer system. Lethally irradiated, thymectomized mice were reconstituted with thymus cells from donors of various ages together with rabbit anti-mouse brain antiserum and complement- (C) treated bone marrow cells. Mice were immunized with TNP-BGG and the magnitude and affinity distributions of their splenic plaque-forming cell (PFC) responses were assayed 3 weeks later. The results indicate that neonatal thymic cells are capable of helping a direct PFC response but cannot mediate the shift to indirect plaque formation. The ability to mediate the switch to an indirect PFC response is a separate maturation event that occurs between birth and 2 to 4 days of age. The thymus cell population from 2-day-old donors is already capable of mediating selection of high affinity PFC. Neonatal cells residing in an irradiated, thymectomized or nonthymectomized adult recipient for 7 days, even in the presence of adult rabbit anti-mouse brain and C-treated syngeneic spleen cells, did not mature to be capable of mediating an indirect PFC response.
T cell requirements for generation of helper factor(s) in man: analysis of the subsets involved.
Authors: Reinherz E L , Kung P C , Breard J M , Goldstein G , Schlossman S F
Published: December 2022

Lymphocyte mitogenic factor functions as a nonspecific helper molecule generated from antigen stimulation of T lymphocytes. As such, it induces proliferation of all major lymphocyte subclasses (T, B, and Null cells) and B cell immunoglobulin synthesis. In the present study, the specific T cell subset requirements for LMF production were determined in man. By utilizing the OKT4 monoclonal antibody directed at the human inducer T cell subset, T lymphocytes were separated into OKT4+ and OKT4- subpopulations. The antigen stimulated OKT4+ subset generated LMF in a fashion comparable to the unfractionated T cell population and was itself capable of directly inducing B cell proliferation and antibody production. In contrast, LMF could not be generated from the OKT4-T cell subset although this population accounted for 40% of the unfractionated T cell population. In addition, the OKT4- subset was unable to facilitate B cell differentiation in the presence of soluble antigen. These studies provide the first report of T cell subset restriction for generation of helper factors in man and further stress the importance of the OKT4+ inducer population in regulation of the human immune response.
Expression of both I-A and I-E/C subregion antigens on accessory cells required for in vitro generation of cytotoxic T lymphocytes against alloantigens or TNBS-modified syngeneic cells.
Authors: Pettinelli C B , Ahmann G B , Shearer G M
Published: December 2022

We have previously reported that radioresistant, Thy 1-negative accessory cells (SAC) are required for the in vitro generation of cytotoxic T-effector cells to allogeneic or trinitrophenyl-modified syngeneic cells. These SAC were found to provide accessory functions irrespective of whether they were syngeneic, semi-syngeneic, or allogeneic to the responding cells. To further characterize the accessory cells active in CML, the expression of Ia antigens on this functional population was assessed by pretreated SAC with anti-Ia reagents and complement and by testing the accessory cell function of these treated populations. The results of these studies demonstrated that the relevant accessory cells for allogeneic and TNP-self CTL express Ia determinants encoded by genes mapping in the I-A and I-E/C subregions. For the TNP-self CTL the accessory function of both SAC syngeneic or allogeneic to the responding and stimulating cells was specifically abrogated by treatment with anti-Ia reagents and complement.
A monoclonal antibody reactive with human peripheral blood monocytes.
Authors: Breard J , Reinherz E L , Kung P C , Goldstein G , Schlossman S F
Published: December 2022

A monoclonal antibody directed at a determinant on human peripheral blood monocytes was produced and characterized. This hybridoma antibody, termed OKM1, was reactive by indirect immunofluorescence and complement- (C) mediated lysis with adherent mononuclear cells. OKM1 was unreactive with lymphocytes, thymocytes, lymphoblastoid cell lines, and tumor cells of the T or B cell lineage. In contrast, acute myelomonocytic leukemia cells and granulocytes were reactive with the antibody. Pretreatment of peripheral blood mononuclear cells with OKM1 and C before culture with soluble antigens totally abolished their antigen-induced proliferative response. This function was restored by addition of 1% adherent cells. These findings provided additional support for the notion that OKM1 was reactive with monocytes. In addition, OKM1 appeared to define two distinct populations of monocytes; an adherent population of large cells bearing surface Ia determinants and a nonadherent population of small, Ia-negative cells. These OKM1+ Ia- cells were found to be a contaminant of most fractionated mononuclear cell subsets including the E-SIg-Null cell population.
Diffusion control of the binding of carcinoembryonic antigen (CEA) with insoluble anti-CEA antibody.
Authors: Frackelton A R , Weltman J K
Published: December 2022

Analysis of the kinetics of CEA binding to rabbit anti-CEA antibody insolubilized on filter paper discs provided six lines of evidence indicating diffusion control of this reaction: 1) The observed forward rate constant, k1obs, decreased when solvent viscosity was increased with sucrose or Ficoll. 2) k1obs per receptor site increased from 1.2 x 10(4) M-1 sec-1 to 2.6 x 10(4) M-1 sec-1 as receptor site density on immunosorbent discs decreased from 5.2 x 10(-13) to 2.1 x 10(-13) moles/disc. 3) Stirring the reacting mixture of CEA and insoluble anti-CEA increased k1obs from 1.2 x 10(4) M-1 sec-1 to 3.4 x 10(4) M-1 sec-1. 4) The activation energy for CEA-insoluble anti-CEA binding was indistinguishable (p greater than 0.1 by t-test) from 4.1 Kcal/mole expected for diffusion controlled reactions. 5) Bimolecular reaction theory predicted a diffusion limited forward rate constant, k1max, for CEA binding to insoluble anti-CEA, which was consistent with our observed k1 of 1.2 x 10(4) M-1 sec-1. 6) k1obs for CEA binding to soluble rabbit anti-CEA antibody was 6.4 x 10(4) M-1 sec-1, 5 times faster than the k1obs of the heterogeneous phase reaction for receptor site density of 5.2 x 10(-13) moles/disc. Diffusion control of ligand binding to insoluble receptors, as exemplified by the CEA-insoluble anti-CEA model system, may be a very general biologic phenomenon.
The formation and release of slow reacting substance of anaphylaxis (SRS-A) by rat and mouse peritoneal mononuclear cells induced by ionophore A23187.
Authors: Orange R P , Moore E G , Gelfand E W
Published: December 2022

The capacity of the calcium ionophore A 23187 to induce the formation of SRS-A was used in an attempt to identify the nature of the cells that are responsible for the formation of this material. In agreement with previous observations, rat and mouse peritoneal mast cells were found to contribute very little to the total amount of SRS-A that was made, and in fact their removal from the cell suspensions resulted in an augmentation of the amount of SRS-A that was found. Discontinuous Ficoll and bovine serum albumin gradients were employed to further characterize these SRS-A-releasing cells from induced peritoneal cells. SRS-A was produced by nonspecific esterase-positive and latex-phagocytizing mononuclear cells. By electron microscopy, these cells had macrophage-like characteristics. In contrast, rat alveolar macrophages and a macrophage-like cell line of mouse origin both failed to produce large amounts of SRS-A. The results suggest that SRS-A formation may be the property of an as yet imperfectly characterized subclass of macrophages.