Mapping of the major histocompatibility complex and viral antigens on the plasma membrane of a measles virus-infected cell

The two measles virus glycoproteins, the hemagglutinin and fusion protein, are expressed on the surfaces of infected cells. Although the two molecules are chemically distinct, they associate on the cell surface, judging from their ability to comigrate (co-cap). However, neither is directly complexed with the major histocompatibility (MHC) gene products, HLA-A, -B, -C or -D, on the plasma membrane, based on results from three distinct assays. First, in tests of capping, these viral glycoproteins failed to comigrate with any HLA determinant. Second, electron microscopy showed that the viral glycoproteins occupied domains on the plasma membrane distinct from MHC gene products; 125I labeling of cell surface determinants and subsequent analysis by immune precipitation and PAGE confirmed this result. Third, incubation of measles virus-infected cells in the presence of monoclonal or polyclonal antibodies to measles virus glycoproteins removed the viral glycoproteins from the cells' surfaces but did not cause a corresponding decrease in amounts of HLA molecules. These results indicate that the hemagglutinin and fusion polypeptides of measles virus lie in close association on the plasma membrane; however, neither is linked with MHC gene products.     

The effect of histocompatibility antigens on lymphocyte migration in the mouse

Migration patterns of ⁵¹Cr-labelled murine lymph node cells were studied in syngeneic, allogeneic, semi-allogeneic and congenic strain combinations. In certain allogeneic and semi-allogeneic combinations with strong H-2 histoincompatibility, reduced migration of a donor population representing recirculating lymphocytes into recipient lymph nodes was observed. This phenomen could be reproduced in congenic strains differing only at the H-2 locus, and was not seen in congenic strains differing at the Ly-A, Ly-B, RZ, M, or θ loci.

The kinetics of reduced lymphocyte homing to allogeneic lymph nodes and the role of host and donor recognition were investigated. These studies are discussed in relation to possible mechanisms of action, and relevance to factors regulating lymphocyte recirculation.

     

Xenogeneic recognition of tumour specific plasma membrane antigens derived from mouse lymphoma cells

Rabbit antisera were prepared to two mouse ascitic lymphomas, one of them being highly immunogenic, the other one weakly immunogenic in syngeneic hosts. A comparison was made between antisera raised with intact cells and antisera raised with cell-free fractions composed of purified large plasma membranes. The antisera were tested in vitro for their lytic capacity by the ⁵¹Cr-release technique, and in vivo for their capacity to protect syngeneic mice against a tumour challenge. The antisera were used unabsorbed, absorbed with syngeneic mouse tissue, and also after passage through syngeneic animals. The antisera with the highest specificity for the respective tumour in mice were those raised by plasma membranes derived from the strongly immunogenic tumour. Lysis in the ⁵¹Cr test corresponded roughly with these findings. Xenogeneic recognition of tumour distinct antigens as well as the therapeutic value of xenogeneic antisera against established lymphomas growing in the ascitic form are discussed.     

Interactions of interferons in the induction of histocompatibility antigens in mouse fibroblasts and glial cells

The interactive effects of interferons (IFN) in the induction of class II major histocompatibility complex (MHC) antigens in a murine fibroblast line and murine glial cells (primary astrocytes and an oligodendroglioma line) were examined. It was found that IFN-alpha and -beta were additive with IFN-gamma in the induction of class I antigens but that both IFN-alpha and -beta down-regulated the induction of class II antigens by IFN-gamma. This was most clear cut when the IFN-alpha or -beta was present in large excess in terms of anti-viral activity. Recombinant IFN-alpha 2 was found weakly to induce class II antigens, unlike natural IFN-alpha. The results imply that IFN-alpha and -beta may be important in the control of class II antigen expression in vivo although not by themselves inducing class II antigens.     

Thyrogastric autoimmune disease. Studies on the cell-mediated immune system and histocompatibility antigens.

Cell-mediated immune responses were studied in autoimmune diseases of thyrogastric type, Hashimoto's thyroiditis and autoimmune pernicious anaemia-type gastritis. Specific cell-mediated immunity was investigated by the leucocyte migration inhibition procedure, and general cell-mediated immunity (T-cell performance) was studied by standard in vivo and in vitro tests. In thyrogastric autoimmune diseases inhibition of migration of leucocytes was induced by thyroglobulin and gastric parietal cell microsomes; under conditions of presumably low cellular sensitization, stimulation of migration was observed. There was no depression of general cell-mediated immunity, in contrast to what occurs in systemic lupus erythematosus and related autoimmune diseases. A weak association of autoimmune gastritis with HL-A3 and HL-A7 (P LESS THAN 0.05) lost significance when an appropriate correction was applied; this weakness with HL-A clearly does not explain the strong genetic component in thyroid and gastric autoimmunity.     

Action of vasopressin on ATPase activity of microsomal fractions of rabbit heart and liver

Intravenous injection of vasopressin in a dose of 5 pressor units/kg body weight led after 1 h to changes in the ATPase activity of rabbit heart and liver microsomes. These changes differed in direction: Mg- or Ca-activated ATPase activity of the cardiac microsomes was very slightly increased, whereas ATPase activity of the hepatic microsomes was reduced.Laboratory of Molecular Pathology and Biochemistry, Institute of General Pathology and Pathological Physiology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. M. Chernukh.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 12, pp. 1433–1434, December, 1976.     

Differences in the expression of histocompatibility antigens on mouse lymphocytes and tumor cells: immunochemical studies.

Immunochemical studies have shown that labeled, detergent-solubilized extracts of SL2 (H-2d) lymphoma cells contain components reactive with several anti-H-2 alloantisera of restricted specificity. Anti-H-2k and anti-H-2ja as well as anti-H-2d sera precipitated labeled polypeptides of a molecular weight similar to that of H-2 heavy chains. In addition, all antisera tested precipitated a component of 70000 daltons molecular weight, which is antigenically related to gp 69/71 of Friend murine leukemia virus. Reactions with antisera directed against haplotypes other than H-2d could be blocked by addition of unlabeled, detergent-solubilized extracts of H-2d lymphocytes, or by H-2 antigens against which the antiserum was directed. Sequential immunoprecipitations initially using antisera against the K, D, or L region gene products to remove individual known H-2d antigens have made possible the identification of some molecules responsible for these reactions. The results show that antisera against haplotypes other than H-2d which react with SL2 cells, cross-react with normal H-2d antigens. Quantitative absortion of these antisera with intact or solubilized cells has shown that lymphocytes and tumor cells differ in their expression of some H-2 determinants. The antibodies bind only weakly to intact H-2d lymphocytes, but strongly to the corresponding detergent-solubilized antigens. These results do not, therefore, support the derepression hypothesis put forward earlier.     

The B-L (Ia-like) antigens of the chicken. Lymphocyte plasma membrane distribution and tissue localization

Specific antisera reacting with B-L (Ia-like) antigens were prepared by reciprocal immunization of animals from the congeneic lines CB and CC. The resulting antisera were tested either in direct or indirect immunofluorescence tests and stained 10-16% of peripheral blood cells (PBL). Of the B-L+ cells, 90% were B cells and 8% were T cells. After in vitro stimulation of PBL with ConA, 58% were B-L+ and 91% of these were T cells. B and T cells were defined by means of rabbit antisera raised against bursa and thymus cells made specific by absorption with the relevant tissues. Antigens determined by anti-B-L antisera, rabbit anti-bursa (ABS) and rabbit anti-thymus (ATS) sera showed an independent distribution on the membrane of PBL. The tissue distribution of B-L+ cells, defined by means of allo-antisera and monoclonal antibodies, was studied by direct and indirect immunofluorescence on sections of skin, liver, kidney and brain. In all organs, in addition to B cells and a small number of, presumably activated, T cells, macrophages and dendritic cells were positive. Notably, glia cells in the brain were also shown to express B-L antigen.     

Immunochemical comparison of synaptic plasma membrane and synaptic vesicle membrane antigens

Summary A synaptic vesicle fraction and a synaptic plasma membrane fraction obtained after subfractionation of synaptosomes from chick forebrain have been used to produce antisera in rabbits.Immunofluorescence histology with the two antisera revealed that they reacted strongly with synaptic terminal regions present in the chick forebrain, cerebellum and spinal cord. In addition, the synaptic plasma membrane antiserum (but not the synaptic vesicle antiserum) reacted with preterminal axons in the cerebellum and spinal cord.Comparison of the two antisera by two-dimensional immunoelectrophoresis, revealed the presence of common antigens in the synaptosomal vesicle and plasma membrane fractions.Incubation of synaptosomesin vitro with the synaptosomal vesicle antiserum and complement produced a dose-dependent inhibition of synaptosome swelling up to a maximum of 55% of that obtained with the synaptosomal plasma membrane antiserum. The results of this test are consistent with the hypothesis that some synaptosomal vesicle antigens may be present also in the synaptosomal plasma membrane and imply that they face the external surface of the synaptosomes.The fate of vesicle membrane components in synaptosomal plasma membranes is not known. The possibility is discussed that they may be recycled locally by a mechanism similar to that proposed by Heuser and Reese (1973) for re-use of synaptic vesicle membranes at the neuromuscular junction.     

Tumorigenicity, major histocompatibility antigens, and karyotypes of interspecific hybrids between mouse neuroblastoma and rat glioma or liver cells

Five interspecific hybrids of mouse neuroblastoma with rat glioma (NG108-15, 140-3, and 141-B) or with nontransformed rat liver cells (NBr-10A and NBr-20A) were examined for major histocompatibility (MHC) antigens and tumorigenicity in comparison with their karyotypes. Both mouse and rat MHC antigens were present in each hybrid population, as determined by a simple cytotoxicity test. All five hybrid cell lines produced tumors in athymic nude mice with varied take incidences. Four hybrid cells, NG108-15, 140-3, NBr-10A, and NBr-20A, were highly tumorigenic. Their karyotypes were characterized by a higher modal chromosome numbers than would be expected from the fusion of parent cells in which at least one parent contained an increased number of chromosomes. In contrast, 141-B cells, with massive loss of chromosomes from both malignant parents, were weakly tumorigenic. The results suggest that the retention of marker chromosomes as well as double minutes (DMs) or microchromosomes of neuroblastoma origin may be required for expression of malignancy in these hybrid cells. The survival time of tumor-bearing mice also varied within the five cell lines, but it was significantly short in NG108-15, which yielded lung metastases in the host animals.     

Enhancement of the expression of histocompatibility antigens of mouse lymphoid cells by interferon in vitro

The expression of surface antigens of thymocytes and splenic lymphocytes was determined by quantitative absorption of alloantibodies. Mouse interferon preparations enhanced the expression of histocompatibility antigens of thymocytes from mice of different ages, but of splenic lymphocytes only from young mice. Interferon did not affect the expression of the theta antigen of thymocytes or splenic lymphocytes.     

Autoantibodies,histocompatibility antigens and testosterone in males with alcoholic liver cirrhosis.

Titres and immunoglobulin classes of autoantibodies were examined in 69 male patients with alcoholic liver cirrhosis and the findings were related to particular human leucocyte antigens and serum concentration of testosterone. Both anti-nuclear antibodies (ANA) and smooth muscle antibodies (SMA) were significantly more prevalent in patients with cirrhosis than in sex- and age-matched controls. Antimitochondrial antibodies and liver cell membrane antibody were found in 4% of the patients, and in none of the controls, but this difference was not significant. Patients with HLA-B8 and/or HLA-B12 had higher titres of ANA (n.s.) and SMA (P less than 0.05) than patients without these HLA antigens. Serum concentrations of testosterone were significantly lower in ANA-positive patients than in those negative (P less than 0.05), and a similar tendency was found in SMA-positive patients. With increasing titres of ANA the concentration of testosterone fell. Serum concentration of testosterone correlated inversely (P less than 0.05) with plasma immunoglobulin G and A. It is concluded that both genetic and hormonal factors may influence the humoral immune response in these patients.     

Age-associated oxidative damage in microsomal and plasma membrane lipids of rat hepatocytes

Phosphatidylcholine hydroperoxide (PC-OOH) and phosphatidylethanolamine hydroperoxide (PE-OOH) concentrations were determined in microsomes and plasma membranes prepared from 2- and 17-month-old male Sprague-Dawley rat hepatocytes, to verify the dissimilarity of age dependency of lipid peroxidation in organelle membranes. The hydroperoxides were directly measured by chemiluminescence detection–high-performance liquid chromatography (CL–HPLC), and 1-palmitoyl-2-(13-hydroperoxy-cis-9, trans-11-octadecadienoyl) phosphatidylcholine (PLPC-OOH) and 1-palmitoyl-2-(13-hydroperoxy-cis-9, trans-11-octadecadienoyl) phosphatidylethanolamine (PLPE-OOH) were enzymatically synthesized and utilized as standards for the calibration. Baseline concentrations of hydroperoxides (PC-OOH+PE-OOH) of the 17-month-old rats were 46 pmol per mg protein in microsomes (2.7 times higher than the 2-month-old rats) and 306 pmol per mg protein in plasma membranes (9.9 times higher than the 2-month-old rats). Both microsomal and plasma membrane lipids were severely peroxidized and converted to phospholipid hydroperoxides by NADPH-dependent lipid peroxidation in vitro, but the age-dependency was only observed in the plasma membranes. These results demonstrate that substantial oxidative damage to membrane phospholipids occurs with ageing both in microsomes and plasma membranes, but is more prevalent in plasma membranes in rat hepatocytes.