bluuebunny/crossref_metadata_2025 huggingface dataset:
Purification of a major membrane protein of Toxoplasma gondii by immunoabsorption with a monoclonal antibody.
Authors: Kasper L H , Crabb J H , Pfefferkorn E R
Published: December 2022

The principal iodinatable surface protein (P30) of our cloned RH strain of Toxoplasma gondii has an apparent molecular weight of 30,000, as measured by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate under reducing conditions. Monoclonal antibody B specifically immunoprecipitated protein P30 from a detergent extract of surface radioiodinated T. gondii. Monoclonal antibody B in the presence of complement was also parasiticidal for T. gondii, and this parasiticidal effect could be blocked by protein P30. Monoclonal antibody B was purified from mouse ascitic fluid and linked to cyanogen bromide-activated Sepharose. The resulting immunoabsorbent was used to purify 1.7 mg of protein P30 from a large number of parasites. The efficiency of recovery of protein P30 was measured by assays of radioactivity and of parasiticidal blocking activity. Protein P30 represented 3 to 5% of the total protein. It is also present in a recently isolated strain of T. gondii. A convalescent human antitoxoplasma serum immunoprecipitated radiolabeled protein P30. Three convalescent antisera when quantitated by an ELISA test had a high anti-protein P30 titer. Charge shift electrophoresis showed that protein P30 has an extensive hydrophobic region and thus is probably an integral membrane protein. Electrophoresis under nonreducing conditions showed no evidence that protein P30 exists as a disulfide linked homo- or heterodimer, although it probably has intramolecular disulfide bonds.
Analysis of a defect in the H-2 genes of SV40 transformed C3H fibroblasts that do not express H-2Kk.
Authors: Rogers M J , Gooding L R , Margulies D H , Evans G A
Published: December 2022

C3H fibroblasts transformed in vitro with SV40 were adapted to in vivo growth. Several clones were isolated from a single, highly oncogenic tumor and those that displayed oncogenic potential also no longer expressed the H-2Kk molecule. Using the technique of Southern blot hybridization, the H-2 genes and integrated SV40 sequences present in the genomic DNA of several of these clones have been examined and compared with both the parent line and normal liver genomic DNA from C3H mice. All H-2Kk negative clones had altered H-2 genes that appeared as a gain and, depending on the restriction endonuclease, loss of hybridizing fragments compared to normal C3H DNA. A 5.5-kb fragment missing from the Sstl digests of the H-2Kk negative variants was mapped to the H-2Kk region of the major histocompatability complex with the use of congenic mice. This provided direct evidence that a mutation had occurred in the H-2Kk region. The integrated SV40 sequences were similar to those already seen in other SV40 transformed cells and not closely linked to any of the H-2 genes. There was no indication that the H-2 mutation was caused by integration of SV40.
A common VH marker relating BALB/c alpha 1-3 dextran-binding and A/J P-azophenylarsonate-binding antibody families.
Authors: Stanislawski M , Pene J
Published: December 2022

Antibodies that define the idiotypic marker 104E-MPC11 were purified from rabbit anti-M104E sera. The expression of this marker was searched for in a panel of representative myeloma and hybridoma proteins differing in antigen-binding specificities and idiotype expression. By radioimmune competition assays, the marker was distinguished from the IdX Dex idiotype of BALB/c alpha 1-3 dextran-binding proteins (alpha 1-3 Dex-BP) in that it was expressed on both IdX Dex-positive and IdX Dex-negative proteins on the one hand, and on A/J p-azophenylarsonate-binding proteins (Ar-BP) on the other. Among the later, six of eight of the proteins also expressed the major cross-reactive idiotype (CRI). The marker was expressed weakly or not at all on A/J Ar-BP, which did not express the CRI, as well as on a number myeloma and hybridoma proteins of different antigen-binding specificities, and normal IgG. The marker was localized in VH, but its full expression required the light chain, and was influenced by quaternary structure.
Blocking of CTL-mediated killing by monoclonal antibodies to LFA-1 and LYT-2,3. I. Increased susceptibility to blocking after papain treatment of target cells.
Authors: Gromkowski S H , Heagy W , Sanchez-Madrid F , Springer T A , Martz E
Published: December 2022

It is now established that monoclonal antibodies (MAb) against LFA-1 and Lyt-2,3 antigens on cytolytic T lymphocytes (CTL) block killing function in the absence of C. It has been suggested that the blocking is inversely related to CTL-target affinity. In this report, we studied the effect of papain pretreatment of target cells, because papain is known to remove H-2 and to render target cells more resistant to allospecific CTL. CTL-target conjugate formation was weaker with papain-treated target cells (based on reduced post-dispersion lysis in dextran-containing medium). The concentration of MAb required to produce 40 to 60% inhibition of 51Cr release (2-hr assay) was reduced four to 29-fold for alpha LFA-1 and 64 to 114-fold for alpha Lyt-2,3. Papain, however, did not induce blocking by MAb to other CTL antigens such as Thy-1, H-2, and T200. Flow cytometric analysis confirmed that papain selectively removed more than 95% of H-2. In kinetic studies of removal and recovery, H-2 density and conjugate formation correlated well with each other. Sensitivity to blocking was not as well correlated, raising the possibility that an unidentified papain-sensitive target cell molecule other than H-2 plays an important role in CTL-target interaction.
Cell cycle-dependent expression of antigen-binding and I-J-bearing molecules on suppressor T cell hybridomas.
Authors: Takabayashi K , Suzuki N , Kanno M , Imai K , Tokuhisa T , Tomioka H , Taniguchi M
Published: December 2022

The I-J and antigen-binding chains with constant region determinant (Ct) that compose an antigen-specific suppressor T cell factor were found on the surface of suppressor T cell hybridomas, serologically and morphologically demonstrated by a fluorescence-activated cell sorter (FACS) and immunoelectron microscopic analyses. Moreover, the surface expression of the I-J and Ct-bearing chains fluctuating with the same kinetics depended entirely upon the cell cycle. The maximum expression of these two chains was observed in the early stage of the M phase, and the minimum in the S phase. Similarly, the magnitude of the suppressor activity was maximal in the late stage of the M phase, and was minimal in the S phase. The results therefore demonstrated that there exists good correlation between the cell surface expression of the I-J and Ct-bearing chains and the magnitude of the suppressor activity produced. The antigen recognition units on suppressor T cell hybridomas have serologically and morphologically been characterized by using radiolabeled antigens or monoclonal antibodies against the I-J or Ct on the antigen-binding molecule. Cell-binding assay and radioautographic analysis demonstrated that the suppressor T cell hybridoma possesses the capacity to bind native antigen in an antigen-specific fashion as does the hybridoma-derived, antigen-specific suppressor factor composed of the I-J and the Ct-bearing chains, indicating that the recognition unit on the cell surface is composed of a structure similar to the factor.
Immunohistologic and immunoelectron microscopic characterization of the mucosal lymphocytes of human small intestine by the use of monoclonal antibodies.
Authors: Cerf-Bensussan N , Schneeberger E E , Bhan A K
Published: December 2022

Monoclonal antibodies reactive with T cells, T cell subsets, B cells, monocytes, and natural killer cells were used to characterize the nature of mucosal lymphocytes in the human small intestine by application of the immunoperoxidase technique to tissue sections for light and electron microscopic examination. In addition, for comparison, peripheral blood mononuclear cells (PBL) were studied by immunoelectron microscopy. Most of the intraepithelial lymphocytes (IEL) were T cells (Leu-1+, T3+) and expressed the phenotype associated with cytotoxic/suppressor T cells (Leu-2a+, T8+). In contrast, a majority of T lymphocytes in the lamina propria expressed the phenotype associated with helper/inducer T cells (Leu-3a+, T4+). These observations confirm and extend the findings previously reported. In addition, a small number of cells in the lamina propria with the ultrastructural features of macrophages were found to react with anti-Leu-3a and anti-T4 antibodies. Although many IEL contained cytoplasmic granules and had ultrastructural features similar to those of circulating granular lymphocytes, none of these cells reacted with anti-Leu-7 (HNK-1), anti-T10, or anti-M1 antibodies. This suggests that IEL may not be related to circulating large granular lymphocytes, which are Leu-7+, T10+, M1+ and are associated with natural killer activity. Not only Leu-7+ PBL, but T8+, T4+, or T3+ mucosal lymphocytes or PBL also may contain cytoplasmic granules. Therefore, the cytoplasmic granules are not restricted to one cell type, in particular, to Leu-7+ cells.
Immunologic effects of interleukin 2 in primary immunodeficiency diseases.
Authors: Flomenberg N , Welte K , Mertelsmann R , Kernan N , Ciobanu N , Venuta S , Feldman S , Kruger G , Kirkpatrick D , Dupont B , O'Reilly R
Published: December 2022

Five children with primary deficiencies of T cell function were studied to assess the effects of highly purified exogenous Interleukin 2 (IL 2) on their in vitro T cell responses. The lymphocytes from one child with Nezelof's T cell deficiency demonstrated absence of endogenous IL 2 production and improved proliferative responses to mitogen or alloantigen in the presence of exogenous IL 2. Moreover, during in vitro mixed lymphocyte culture in the presence of exogenous IL 2, his lymphocytes were able to develop into cytotoxic effector cells. A second child with Nezelof's syndrome demonstrated a different type of defect. The lymphocytes from this child had less impairment of endogenous IL 2 production. Although IL 2 increased the proliferation of his cells in response to PHA, similar augmentation was not seen after stimulation with OKT3 or alloantigen. In cell-mediated cytotoxicity assays, after mixed lymphocyte culture, natural killer-like activity was strongly boosted in the cultures that contained IL 2, but T cell-mediated cytotoxicity was not. The lymphocytes from three patients with severe combined immunodeficiency did not show improved proliferative responses in the presence of IL 2. Thus, only one of the five patients demonstrated the combination of defective endogenous IL 2 production, but preservation of the ability to respond appropriately to exogenous IL 2. This child may therefore have suffered from a T cell defect pathophysiologically similar to that seen in nude or aged mice.
Attenuation of an adjuvant arthritis by type II collagen.
Authors: Trentham D E , Dynesius-Trentham R A
Published: December 2022

Subcutaneous injection of the nonimmunogenic synthetic alkyldiamine, N,N-diotadecyl-N',N'-bis(2-hydroxyethyl) propanediamine (CP-20961), which possesses potent adjuvant properties for cellular sensitization, can induce an inflammatory arthritis in rats. To study whether a host reaction to native type II collagen plays a role in the pathogenesis of the arthritis in this model, CP-20961-injected Lewis rats received i.v. on four occasions native type II collagen coupled to syngeneic spleen cells with ethylcarbodiimide (CDI), native type II collagen added to spleen cells without CDI, or native type II collagen coupled to rat red blood cells (RBC) with glutaraldehyde. There was a significant suppression of the severity of arthritis in all three groups compared with a control group injected with CP-20961 but not receiving cells. In addition, the prevalence of arthritis was decreased in the group receiving native type II collagen-coupled RBC. Injection of cells coupled to denatured type II collagen, native type I collagen, and ovalbumin did not affect significantly the morphologic aspects of this disease. These data provide evidence that material possessing the quaternary epitope(s) of type II collagen functions in an as yet unidentified effector pathway in this adjuvant arthritis.
Production of a human T lymphocyte chemotactic factor by T cell subpopulations.
Authors: Van Epps D E , Potter J W , Durant D A
Published: December 2022

Concanavalin A-stimulated human peripheral blood mononuclear cells release a lymphocyte chemotactic factor. This lymphocyte chemotactic factor is produced optimally after 24 to 48 hr of culture and is not found before 3 hr of culture, which suggests that the factor is synthesized de novo and is not preformed and secreted after Con A stimulation. This is further supported by experiments showing that the protein synthesis inhibitors cycloheximide and puromycin totally prevent the production of the chemotactic factor. Experiments using cultured and uncultured T lymphocytes as responding cells show that cultured T cells respond more efficiently than uncultured T cells to this factor. Furthermore, the lymphocyte chemotactic factor preferentially stimulates T lymphocyte locomotion as compared to peripheral blood non-T lymphocyte migration. Fractionation of mononuclear cells into glass nonadherent lymphocytes, monocyte-enriched preparations, T lymphocytes, and non-T lymphocytes shows that lymphocyte chemotactic factor is produced by Con A-stimulated, glass nonadherent lymphocytes and T cells but not by monocytes or non-T lymphocytes. Further fractionation of T lymphocytes into Leu-2 and Leu-3 T cell subpopulations shows that the production of T lymphocyte chemotactic factor can be attributed to the Leu-2 suppressor/cytotoxic T cell subset. The generation of a T lymphocyte chemotactic factor by Leu-2 T cells may represent a means of recruiting other T cells to the site of its release.
Generation of low m.w., C3-bearing immunoglobulin in human serum.
Authors: Jacobs R J , Reichlin M
Published: December 2022

The generation of low m.w. C3-bearing immunoglobulin (lg) in normal human serum by an immune complex (IC) model was investigated in vitro by using discontinuous sucrose density gradient centrifugation (DGC) and an assay that measures C3-bearing Ig. In this method developed to measure circulating IC, all C3 and C3-bearing material is precipitated from serum by using anti-C3 sera in C3d antibody excess, and immune precipitated, C3-bearing Ig is quantitated by the uptake of 125I-5S-anti-IgG. When plasma from patients with clinically active systemic lupus erythematosus was assayed after DGC, most of the reactive material was low m.w. (7S), rather than greater than or equal to 19S as expected for IC, in agreement with a previous report. Low m.w., C3-bearing Ig was found in normal EDTA plasma after extended storage at -29 degrees C but not after storage at -70 degrees C. Such material was also generated in normal human serum during incubation at 37 degrees C and its generation was stimulated by the addition of an IC model, high m.w., heat-aggregated IgG (HMW-HAIgG). In experiments in which the participation of serum IgG was monitored by the addition of 125I-7S-IgG and 131I-HMW-HAIgG was used as an IC model, low m.w., C3-bearing Ig was generated exclusively from serum IgG and the amount generated was proportional to the concentration of 131I-HMW-HAIgG. No significant decrease in sedimentation of 131I-HMW-HAIgG was observed, but the ability of anti-C3 sera to precipitate 131I-HMW-HAIgG decreased 66% 4 hr after initial C activation. These results indicate that generation of nascent C3b in serum results in its interaction with monomeric serum IgG, producing low m.w., C3-bearing IgG. In addition, the data indicate that circulating IC that activate C have a brief time span during which they can be detected by methods that depend upon the binding of C3.
Multiple splenic lymphoid cell subpopulations regulate H-2 antigen expression on teratocarcinoma cells in vivo.
Authors: Ostrand-Rosenberg S , Cohn A L , Sandoz J W
Published: December 2022

Undifferentiated murine 402AX teratocarcinoma cells do not express MHC antigens when passaged in vitro or in vivo in genetically susceptible host mice. When passaged in vivo in genetically resistant mice, however, the tumor cells become H-2b antigen positive regardless of the H-2 haplotype of the resistant host mouse. The present studies use monoclonal anti-H-2b antibodies to corroborate these earlier findings, which were performed with conventional antisera. Previous studies have established that host bone marrow plus lymphoid cells from resistant primed donors regulate tumor cell H-2b antigen expression. Using bone marrow and mature lymphoid cell reconstitution techniques, the present studies indicate that splenic Ig- cells from genetically resistant host mice are the most efficient lymphoid cell subpopulation in tumor cell H-2b antigen induction. Ig+ spleen cells also reconstitute the capacity to induce teratocarcinoma cell H-2 antigens but are less effective than Ig- spleen cells. Tumor cell H-2 antigen induction in C57BL/6 beige mice is impaired compared to C57BL/6 hosts, which suggests that host NK cells may also be involved in tumor cell H-2 antigen induction. Reconstitution of lethally irradiated resistant hosts for teratocarcinoma cell H-2 antigen expression requires bone marrow plus resistant primed lymphoid cell subpopulations; bone marrow alone is insufficient. These results indicate that multiple splenic lymphoid cell subpopulations requiring a radiosensitive host environment and/or factor for differentiation regulate teratocarcinoma 402AX H-2b antigen expression in vivo in genetically resistant mice.
Immunoperoxidase localization of large granular lymphocytes in normal tissues and lesions of athymic nude rats.
Authors: Ward J M , Argilan F , Reynolds C W
Published: December 2022

The immunocytochemical localization of cells reacting with the OX-8 monoclonal antibody was studied in athymic nude rats by the avidin-biotin-peroxidase complex (ABC) immunoperoxidase technique. By a number of criteria, including morphology and tissue distribution of OX-8-positive cells, it is postulated that the majority of lymphocytes reacting with OX-8 antibody in nude rats are LGL. The cell surface of large lymphocytes reacted with the OX-8 antibody in fixed tissue sections. The tissues with the greatest density and percentages of these reactive cells included the paracortex of lymph nodes in association with interdigitating cell (IDC) hyperplasia, bronchial-associated lymphoid tissue (BALT), the medullary cords and sinuses of lymph nodes, and intestinal epithelium. Ultrastructure of the paracortical areas of IDC hyperplasia revealed granular lymphocytes in close association with IDC. Tissues with the fewest positive cells included bone marrow, B cell areas of lymphoid tissues, and parenchymal epithelial organs. In healthy and cachectic nude rats, various incidental and pathologic lesions contained a significant number of OX-8-positive cells. Large numbers of positive lymphocytes were seen in suppurative pneumonic lesions, enteritis, focal ulcerative epithelial lesions, and papovaviral sialoadenitis. The observation of a large number of OX-8-positive lymphocytes in nonlymphoid organs supports the hypothesis that these cells play a major role in the first line of defense against not only tumors but also infectious agents.
Alteration of T cell subsets and induction of suppressor T cell activity in normal subjects after exposure to sunlight.
Authors: Hersey P , Haran G , Hasic E , Edwards A
Published: December 2022

The effects of exposure to natural sunlight on the immune system were studied in 15 normal human subjects. Exposure was for 1 hr each day for 12 days over 2 wk and tests were carried out before, on completion, and 2 wk after completion. In comparison to concurrent studies on 13 age- and sex-matched controls, sun-exposed subjects had a significant increase in their circulation of T cells recognized by OKT8 monoclonal antibodies and a decrease in OKT4 positive T cells. Suppressor T cell activity measured in pokeweed mitogen-stimulated cultures of T and B cells was significantly increased against IgG and IgM production. These changes were still evident in many of the subjects 2 weeks after completion of the sun exposure. A trend for depression of natural killer cell activity against a melanoma target cell was noted in the present study, but this did not appear as marked as that noted previously in subjects exposed to radiation in solariums. The differences between the effect of radiation from solariums and natural sunlight on the immune system may result from the higher dosage of UV-A in radiation from solariums. The results suggest that exposure to sunlight may favor the induction of suppressor pathways in response to antigenic stimuli and that this may limit immune responses against tumor cells such as melanoma. They support the idea from animal studies that systemic changes in the immune system may be an important factor in the association of UV radiation with malignancy.
Decreased Fc receptor avidity and degradative function of monocytes from patients with systemic lupus erythematosus.
Authors: Katayama S , Chia D , Knutson D W , Barnett E V
Published: December 2022

We studied the binding and degradation of stable, soluble heat aggregates of 125I-IgG (A-IgG) by monocytes from 30 patients with systemic lupus erythematosus (SLE) and 30 normals. Relative avidities (KE) for Fc receptor (FcR) binding of A-IgG and maximal binding of A-IgG by monocytes were determined from Scatchard plots of binding data obtained at 4 degrees C. Rates of degradation (Vmax) of A-IgG at 37 degrees C were calculated from Lineweaver-Burke plots of the Michaelis-Menton equation. KE were decreased in SLE monocytes (15.5 X 10(-9) L/M) as compared with normals (20.1 X 10(-9) L/M, p less than 0.005) and Vmax were decreased for SLE (0.89 ng/hr) as compared with normals (1.11 ng/hr, p less than 0.005). The maximal FcR binding by SLE monocytes was not statistically different in SLE patients and normals, but monocytes from SLE patients with active disease showed a lower maximal binding capacity for A-IgG (4.9 ng/10(5) cells) than normals (5.4 ng/10(5) cells, p less than 0.05). KE and Vmax in SLE were also lower for patients with active disease than for normal subjects. KE in patients whose anti-ssDNA binding was greater than 20% were lower than for those with DNA binding of less than 20% (p less than 0.005). These data suggest that patients with active SLE have diminished numbers of available FcR on their circulating monocytes, possibly due to interiorization of FcR during endocytosis of endogenous circulating immune complexes.
Autoimmunity and immune complex disease after neonatal induction of transplantation tolerance in mice.
Authors: Goldman M , Feng H M , Engers H , Hochman A , Louis J , Lambert P H
Published: December 2022

Mice made neonatally tolerant to alloantigens were found to develop an immunologic disease resembling systemic lupus erythematosus. In BALB/c mice neonatally injected with C57BL/6 X BALB/c F1 hybrid spleen cells, features of autoimmunity were observed first. After 5-24 wk, antinuclear, anti-SS DNA, thymocytotoxic, and rheumatoid factor-like antibodies were detected in association with hypergammaglobulinemia and with the occurrence of circulating immune complexes and cryoglobulins. Some of the antinuclear antibodies were found to be produced by F1 donor B cells persisting in the host. Second, immunopathologic changes were detected in tolerant mice. In the kidneys, an immune complex glomerulonephritis of the membranous type was observed. Immunoglobulin deposits were also found in the choroid plexus and at the dermoepidermal junction. In addition, thrombocytopenia was a common finding, and a positive direct Coomb's test occasionally was detected. Features of autoimmune disease were closely associated with the effective induction of transplantation tolerance, as revealed by the inability of spleen cells to generate in vitro cytolytic responses against C57BL/6 alloantigens. It is suggested that, although transplantation tolerance is associated with a lack of cytolytic reaction of the host against F1 hybrid donor alloantigens, other types of allogeneic interactions could lead in this model to the development of autoimmunity and immunopathology.
Immunologic and biochemical properties of several retinal proteins bound by antibodies in sera from animals with experimental autoimmune uveitis and uveitis patients.
Authors: Gregerson D S , Abrahams I W
Published: December 2022

Sera from guinea pigs and rabbits with and without experimental autoimmune uveitis (EAU) induced by immunization with retina, choroid, optic nerve, retinal rod outer segments (ROS) and purified bovine S-antigen were tested for the ability to immunoprecipitate 125I-labeled, detergent-solubilized bovine retinal proteins. The results demonstrate that three major protein antigens with m.w. of 50,000 (p50), 35,000 (p35) and 27,000 (p27) and several minor activities between 30,000 and 60,000 m.w. are recognized by antibodies from these animals. The p50 component was immunoprecipitated by sera from animals immunized with whole retina homogenate, the high speed supernatant of whole retina homogenate, ROS, and S-antigen, and has been identified as S-antigen in competition experiments. The p35 band appeared when sera were used that were raised against antigen preparations containing membrane-bound retinal protein, i.e., whole retina homogenate, ROS, and washed ROS, and thus appears to be an ROS membrane protein. The p27 band was found when sera raised against ROS, washed ROS, optic nerve and whole retina homogenate were used, suggesting it is a membrane-bound antigen common to ROS and optic nerve. Serum from animals immunized with homologous choroid did not immunoprecipitate a detectable product. S-antigen and p35 were also precipitated by some uveitis patient sera. Because S-antigen is also an ROS protein as is rhodopsin, a putative uveitogenic retinal antigen, ROS appear to be an unusually rich source of autoantigenic proteins. S-antigen was also shown to be synthesized in the retina, and the primary translation product was indistinguishable from purified S-antigen by SDS-PAGE, thus eliminating the possibility that it is derived from or is cross-reactive with the 67,000 m.w. rhodopsin kinase.
The cellular basis for immune interferon production in autoimmune MRL-Ipr/Ipr mice.
Authors: Santoro T J , Benjamin W R , Oppenheim J J , Steinberg A D
Published: December 2022

Disturbances in immune interferon (IFN gamma) activity have been implicated in the development of human systemic lupus erythematosus (SLE) and the spontaneous disease sustained by autoimmune-prone mice. We therefore investigated the cellular basis for IFN gamma production in MRL-Ipr/Ipr mice and examined the relationship between synthesis of interleukin 2 (IL 2) and IFN gamma. In vitro IL 2 and IFN gamma production in 3 to 6-mo-old, autoimmune MRL-Ipr/Ipr and MRL-+/+ mice was compared with that seen in age- and sex-matched, immunologically normal CBA/J mice. 5 X 10(6) spleen cells were pulsed with 5 micrograms of concanavalin A (Con A), and the cellfree supernatant was assayed for IL 2 and IFN gamma activity at various times up to 72 hr. We found that peak levels of IL 2 in MRL mice were less than 10% of those in the CBA/J. Yet, production of IFN gamma by cells from the autoimmune and normal strains was quite comparable. The addition of murine IL 2 to optimally Con A-stimulated cells from the MRL-Ipr/Ipr or normal mice did not affect the subsequent peak production of IFN gamma. Although the primary producers of IFN gamma in cultures of normal mice bear the Lyt-2+ phenotype, the Lyt-1+2- T-cell subset was found to be the principal source of IFN gamma in the aged MRL-Ipr/Ipr. These data suggest that Lyt-1+ cells from MRL-Ipr/Ipr mice may be differentially responsive to the signal delivered by the same mitogenic lectin with respect to lymphokine production and may indicate a distorted commitment of such cells toward production of IFN gamma and repression of IL 2 synthesis. The relationship between hypoproduction of IL 2, this usual source of IFN gamma, and the autoimmune disease sustained by MRL-Ipr/Ipr mice remains unclear.
Interleukin 2 induces gamma-interferon production: participation of macrophages and NK-like cells.
Authors: Kawase I , Brooks C G , Kuribayashi K , Olabuenaga S , Newman W , Gillis S , Henney C S
Published: December 2022

Interleukin 2 (IL 2) has been shown to be a potent stimulator of natural killer (NK) cells. In the present studies, partially purified mouse and human IL 2 preparations were also found to induce interferon (IFN) from mouse spleen cells. By the criteria of sensitivity to treatment at pH 2 and failure to be neutralized by a potent anti-alpha, beta IFN serum, the species of IFN produced was of type gamma. Cooperation between two types of cell, a macrophage and an NK-like cell, was required for IFN production by murine spleen cells treated with IL 2. The requirement for macrophages could be replaced with supernatant obtained by incubating macrophages for 24 hr with lymphokine preparations containing IL 2. Interestingly, mature T cells apparently played no role in the process. Furthermore, the beige (bg/bg) mutation, which severely impairs NK cell lytic activity, had no effect on the ability of NK-like cells to participate in IFN production. Cell fractionation experiments revealed no dissociation between the requirements for augmentation of NK cytotoxic activity and for IFN production, and it is concluded that at least a portion of the NK boosting induced by IL 2-containing preparations is mediated through gamma-IFN.
Effect of vesicular stomatitis virus (VSV) infection on the development and regulation of T cell-mediated immune responses.
Authors: Sy M S , Tsurufuji M , Finberg R , Benacerraf B
Published: December 2022

Infection of mice with vesicular stomatitis virus (VSV) at the time of immunization failed to enhance T cell-mediated immune response to azobenzenearsonate-(ABA) conjugated spleen cells as measured by delayed-type hypersensitivity and by in vitro proliferation and in vitro generation of ABA-specific cytotoxic T cells. However, mice infected with VSV are incapable of responding to signals from suppressor T cells or their soluble factors. Further analysis revealed that VSV infection does not interfere with the induction of Ts-1 or Ts-2 cells. Because infection of Ts-1 or Ts-2 donors had no effect on the subsequent response seen in the recipients of antigen and suppressor T cells, the most likely candidate for the target of VSV infection is therefore the Ts-3 cell or another T cell interacting with Ts-3. This is supported by our observation that it is possible to bypass the VSV effect by providing the recipients of VSV with normal Lyt-2+-bearing T cells.
Isolation of intact Sm/RNP antigens from rabbit thymus.
Authors: Billings P B , Hoch S O
Published: December 2022

A comparison of the immunologically reactive components of the highly conserved Sm and RNP autoantigens from various mammalian tissue sources suggested the complete absence of a major 26K to 27K Sm-specific polypeptide in rabbit thymus extracts prepared by conventional procedures. A simple modification of the solubilization protocol, achieved by sonicating a suspension of commercial rabbit thymus acetone powder in 0.35 M NaCI, gave an extract containing the full complement of immunologically reactive Sm and RNP proteins detectable in other mammalian species. Without further manipulation, extracts were immediately passed through an immunoaffinity column constructed from human SLE IgG with both anti-Sm and anti-RNP reactivities. The proteins of the purified Sm/RNP were recovered in sufficient quantities for direct analysis by protein staining or immunoblot assays. The antigenic polypeptides were recovered intact and consisted of a single 73K RNP-specific species together with Sm-specific proteins of 26K to 27K (a doublet) and 13K. These proteins were easily visible by protein stain as were nonantigenic components of 35K, 32K, 11K, and less than 10K. The same polypeptides were present in affinity-purified Sm/RNP from HeLa cells, although the RNP protein was slightly smaller. The resolution and integrity of the complexes isolated by this simple two-step procedure, requiring less than 4 hr for completion, is remarkable, and the protein composition of the product compares quite favorably with antigens isolated from other sources by considerably more lengthy and laborious procedures.
Human monocytes, B lymphocytes, and non-B lymphocytes each have structurally unique Fc gamma receptors.
Authors: Cohen L , Sharp S , Kulczycki A
Published: December 2022

Human Fc gamma-binding macromolecules were isolated from subpopulations of mononuclear cells by repetitive affinity chromatography. Mononuclear cells, nylon wool-filtered cells, plastic-nonadherent cells, and plastic-adherent cells from normal donors were radiolabeled by using 125I and lactoperoxidase. Washed cells were solubilized in 1% NP-40 buffer containing proteinase inhibitors at 0 degrees C. Fc gamma receptors were purified on human IgG-Sepharose columns by use of the repetitive affinity chromatography procedure. Analysis by SDS-polyacrylamide gel electrophoresis demonstrated only a 52,000 to 58,000 Mr Fc gamma receptor from nonadherent cell populations. Both rosetting and nonrosetting subpopulations of non-B lymphocytes expressed the 52,000 to 58,000 Mr receptor. The predominant Fc gamma receptor isolated from plastic-adherent cells was a 60,000 to 68,000 Mr macromolecule. Cell preparations enriched in B lymphocytes yielded prominent 43,000 Mr Fc gamma receptors. Thus human monocytes, B lymphocytes, and non-B lymphocytes each appear to have structurally distinct and unique Fc gamma receptors.
Generation of biologically active, complement-(C5) derived peptides by cathepsin H.
Authors: Perez H D , Ohtani O , Banda D , Ong R , Fukuyama K , Goldstein I M
Published: December 2022

The thiol proteinase cathepsin H, isolated and purified from rat liver lysosomes, provokes acute inflammation characterized by the accumulation of polymorphonuclear leukocytes (PMN) when injected intracutaneously into newborn rats. We have examined the possibility that the accumulation of PMN at skin sites injected with cathepsin H is due, in part, to generation locally of C-derived chemotactic factors. We have found that cathepsin H acts in a concentration- and time-dependent fashion in whole human (and rat) EDTA-plasma to generate C5-derived peptides with chemotactic activity for PMN. Chemotactic activity was not generated in EDTA-plasma by either heat-inactivated cathepsin H or by a combination of active enzyme and a thiol proteinase inhibitor isolated from rat epidermis. Cathepsin H also acted in a concentration- and time-dependent fashion on isolated (functionally pure) human C5 to yield chemotactic activity for PMN as well as PMN lysosomal enzyme-releasing activity. Whereas 10 ng/ml cathepsin H generated significant chemotactic activity from isolated C5 (1000 CH50 U/ml), 7 to 10 micrograms/ml were required to generate chemotactic activity in whole EDTA-plasma. Cathepsin H not only was capable of generating biologically active, C5-derived peptides, but also was capable of degrading these peptides. Incubation of either whole EDTA-plasma or isolated C5 with high concentrations of cathepsin H (e.g., 25 micrograms/ml and 100 ng/ml, respectively) caused the rapid appearance of chemotactic activity followed by an equally rapid disappearance. PMN accumulated more rapidly in the skin of newborn rats injected with cathepsin H-treated C5 than in the skin of animals injected with cathepsin H alone. These data suggest that generation by cathepsin H of C-derived chemotactic activity contributes to the ability of this enzyme to induce dermal inflammation.
Erythromyeloid tumor cells (K562) induce PGE synthesis in human peripheral blood monocytes.
Authors: Leung K H , Fischer D G , Koren H S
Published: December 2022

Human peripheral blood monocytes were found to spontaneously produce prostaglandin of the E series (PGE) in culture medium (0.5 ng to 3.0 ng/7.5 X 10(5) cells), and the addition of K562 tumor cells enhanced the production by five- to 15-fold after 18 hr of incubation. PGE2 (10(-6) M) inhibited the cytolytic activity of freshly isolated peripheral blood monocytes against K562 target cells by 50%. The PGE production was inhibited by inhibitors of cyclo-oxygenase (indomethacin, aspirin, and ETYA) when present during the incubation. However, pretreatment of monocytes with these cyclo-oxygenase inhibitors was ineffective in preventing PGE production. Kinetic experiments showed that appreciable stimulation of PGE production occurred only after 6 hr of co-culture. Other human tumor cell lines (HSB, SB, and CEM) enhanced PGE production upon co-culture with monocytes but to a lesser extent (twofold to threefold). Monocytes treated with 0.4% formaldehyde or heat (56 degrees C) were not capable of producing PGE when cultured alone or with K526 tumor cells. In contrast, formaldehyde-treated, but not heat-treated, K562 tumor cells were able to induce monocytes to produce PGE. By using a single cell conjugation assay, K562 tumor cells were found to bind equally well to treated or untreated monocytes. In contrast, the lytic activity of treated monocytes against K562 target cells was abolished. The presence of protein synthesis inhibitor, cycloheximide, was found to inhibit PGE production by monocytes cultured alone or with K562 tumor cells. Supernatants from K562 tumor cell cultures were also capable of inducing monocytes to produce PGE, and their effect on PGE production from monocytes was suppressed by cycloheximide. In addition, pretreatment of either K562 tumor cells or monocytes with an irreversible protein synthesis inhibitor, emetine, also suppressed the production of PGE upon co-culture with the untreated counterpart. The production of PGE by monocytes in response to exposure to tumor cells may represent a mechanism whereby tumor cells subvert host immune defense against them.
Augmentation of NK cell activity by soluble and particulate fractions of Toxoplasma gondii.
Authors: Hauser W E , Sharma S D , Remington J S
Published: December 2022

The ability of sonicates of Toxoplasma gondii and of subcellular fractions of the organism to enhance natural killer (NK) cell activity in mice was examined. Treatment of BALB/c mice with the sonicate preparations enhanced peritoneal and splenic NK activity to the level obtained with intact tachyzoites. Similar augmentation of NK activity in A/J, C57BL/6, and C3H/HeJ mice was also observed. Administration of Toxoplasma sonicate intraperitoneally, intravenously, or subcutaneously enhanced both peritoneal and splenic NK activity. Treatment of mice with a sonicate of L929 fibroblast failed to enhance their NK activity. Concentrations of sonicates ranging from 400 to 4 micrograms of protein enhanced both peritoneal and splenic NK activity. Both the soluble and particulate fractions of the sonicate obtained by differential centrifugation were capable of enhancing peritoneal NK activity, whereas splenic NK activity was enhanced only by the particulate fraction. Heat, protease, and sodium periodate treatment of the Toxoplasma sonicate completely abolished its capacity to augment NK activity. Gel filtration studies revealed that the NK augmenting activity of the Toxoplasma sonicate is distributed over a wide molecular weight range.
Monoclonal antibodies that distinguish non-small cell from small cell lung cancer.
Authors: Mulshine J L , Cuttitta F , Bibro M , Fedorko J , Fargion S , Little C , Carney D N , Gazdar A F , Minna J D
Published: December 2022

Two murine IgG2Ak monoclonal antibodies (703D4, 704A 1) were produced and characterized after immunization with a human large cell lung cancer line (NCI-H 157). These antibodies detect different epitopes on 31 kilodalton [35S]methionine incorporating protein(s). Radiobinding and immunohistochemical studies show these antibodies bind to most (11/13) human non-small cell lung cancer (adenocarcinoma, epidermoid, and large cell), but not to small cell lung cancer (0/11) tumors tested. The epitopes these antibodies recognized are also expressed on human melanomas (7/8), two other tumors (osteogenic sarcoma, renal cell carcinoma), but not on many other human tumors (breast, colon, neuroblastoma, lymphoid), and not on a panel of normal adult human tissues. Because the antigen(s) are preserved after fixation and because of their ability to distinguish lung cancer types from each other and normal tissues, they should be of clinical, as well as of biologic interest.
Depression of natural killer cytotoxicity after in vivo administration of recombinant leukocyte interferon.
Authors: Maluish A E , Ortaldo J R , Conlon J C , Sherwin S A , Leavitt R , Strong D M , Weirnik P , Oldham R K , Herberman R B
Published: December 2022

One hundred thirty-four patients with a variety of malignancies were treated in two phase I clinical trials of recombinant leukocyte A interferon (IFL-rA) produced by recombinant DNA methodology. IFL-rA was given by intra-muscular injection either twice daily or three times weekly for 28 days. Extensive monitoring of natural killer (NK) cell cytotoxicity was done on these patients with rigorously standardized assays and determination of the inherent variability of NK function for each individual. Interferon, as used in these two treatment regimens, failed to produce an appreciable increase in NK activity; in the majority of patients, there was no significant change in NK activity from the pretreatment baseline levels. An unexpected finding was the depression of NK activity in 30% of the patients. The data have been analyzed in terms of their possible relationship to dose and schedules of IFL-rA administration and to antitumor response.
In vivo role of natural killer cells: involvement of large granular lymphocytes in the clearance of tumor cells in anti-asialo GM1-treated rats.
Authors: Barlozzari T , Reynolds C W , Herberman R B
Published: December 2022

The present study was performed to further evaluate the possible in vivo involvement of natural killer (NK) cells in host resistance against tumors. Selective depression of NK activity in Wistar Furth rats was induced by i.p. or i.v. injection of rabbit anti-asialo GM1. This antiserum has previously been shown to produce a decrease in NK activity and a parallel increase in tumor growth in mice. In the present study, rats treated with this antibody showed a parallel decrease in NK activity and in the frequency of large granular lymphocytes (LGL) in the spleen and peripheral blood, indicating that the antiserum-induced depression of NK activity in these sites was probably caused by an elimination of most effector cells. To further determine the possible role of rat LGL in tumor rejection in vivo, we studied LGL involvement in the rapid clearance of radiolabeled tumor cells from the lungs, an assay previously shown to correlate well with in vitro NK activity. Animals treated with anti-asialo GM1 antiserum were found to have a substantial decrease in the in vivo rate of clearance of tumor cells from the lungs. Furthermore, the adoptive transfer of a highly enriched population of LGL into NK-depressed animals 2 hr before tumor challenge, partially restored their cytotoxic activity against established cell lines in vitro and their ability to eliminate radiolabeled cells from the lungs. These results provide direct support for the hypothesis that NK cells are involved in in vivo resistance to tumors, particularly in the elimination of potentially metastatic tumor cells from the circulation and capillary beds.
Two classes of bystander B cell response: activation requirements reflect those of B cells in general.
Authors: Ratcliffe M J , Julius M H
Published: December 2022

Bystander anti-sheep red blood cell (SRBC) antibody responses were induced in cultures containing unprimed B cells, SRBC, particulate or soluble eliciting antigen, and helper T (Th) cells primed to the eliciting antigen. The use of long-term cultured Th cells, depleted of alloreactive cells, allowed direct assessment of the requirements for major histocompatibility complex- (MHC) restricted interactions in the cellular events leading to bystander B cell activation. H-2 restricted Th cell activation limited all bystander B cell responses; however, the requirement for H-2 restricted Th-B cell interactions varied with the state of activation of the responding B cell population before assay. Thus, we defined two classes of bystander B cell response. Activation of small (i.e., resting) SRBC-specific bystander B cells, purified by density gradient centrifugation, was completely restricted by B cell MHC. The requirement for high concentrations of eliciting antigen suggests that nonspecific binding of soluble or particulate antigen to the responding B cell surface plays a mandatory role leading to the activation of resting bystander B cells. The concomitant requirement for a restricted Th-B cell interaction suggests that this role is one of focusing antigen-specific, H-2 restricted T cell help to the resting B cell. In contrast, bystander responses, unrestricted by B cell MHC, were generated exclusively from large (i.e., blasted) B cells limited only by Th cell activation. This class of SRBC-specific bystander response correlated with an SRBC-dependent increase in the number of cells secreting antibodies that recognized degraded mouse RBC. Thus, we propose that the H-2 unrestricted anti-SRBC response is derived from the amplification of B cells activated in vivo to degraded homologous RBC. Both classes of anti-SRBC response were enhanced by the presence of the bystander antigen. This phenomenom is discussed in terms of a mitotic advantage provided by the interaction of antigen with surface immunoglobulin on specific bystander B cells.
Three different signals are required for the induction of cytolytic T lymphocytes from resting precursors.
Authors: Kanagawa O
Published: December 2022

The requirement for the signals in induction of cytolytic T lymphocytes (CTL) has been investigated. C57BL/6 X CBA/T6 F1 spleen cells stimulated with the lectin leukoagglutinin (L-A) failed to show CTL activity in a PHA-facilitated assay, although L-A-activated splenic T cells were able to respond to T cell growth factor (TCGF). Concanavalin A (Con A) on the other hand was able to induce cytolytic activity from CTL-P, as well as to render splenic T cells responsive to TCGF. Furthermore, L-A-activated splenic T cells could generate cytolytic activity upon subsequent culture in secondary mixed leukocyte culture supernatant (2 degrees MLC SN). In contrast, EL-4-derived SN (EL-4 SN) was unable to induce cytolytic activity from L-A-activated spleen cells. In addition, proliferation of L-A-activated spleen cells cultured in EL-4 SN was similar to those cultured in 2 degrees MLC SN. Nonactivated spleen cells were totally unresponsive to both SN in proliferation and generation of CTL. Analysis of T cell clones for the production of a factor necessary for induction of cytolytic activity revealed that both cytolytic and noncytolytic T cell clones were able to produce a factor(s) for the generation of cytolytic activity from L-A-activated T cells. On the other hand, SN from certain antigen-stimulated T cell clones produced factors capable of inducing cytocytic activity by L-A-activated T cells only in the presence of EL-4 SN. Neither EL-4 SN nor cloned T cell SN alone had such a capacity. The nature of the necessary lymphokines in the SN from the clone cells or from the EL-4 is unknown. In the case of the EL-4 SN, it is not known whether the presence of TCGF plays a role or whether that role is perhaps more differentiative than proliferative. This study provides evidence that the induction of CTL from CTL-P can be dissociated into activation, which is required to render T cells responsive to second signals, and differentiation, which is mediated by two different factors.