Molecular mass analysis of murine immunosuppressive immunoglobulin G-binding factors (IgG-BFs) produced by T-cell hybrids

Induced and constitutive murine IgG-binding factors (IgG-BFs) have been purified by affinity chromatography from supernatants of T-cells preincubated with or without murine monoclonal IgG1 and IgG2b, respectively. IgG-BF Mr values have been studied by SDS-polyacrylamide gel electrophoresis (PAGE) after treatment with SDS under conditions which do not noticeably alter their immunosuppressive activities on the secondary in vitro IgG antibody response. Suppression was recovered at Mr values of 80000, 40000 and 20000. When induced IgG-BF was tested, the isotype-specific suppressive activity was found only at 40 kDa. The 20-kDa moiety appeared to derive from the 40-kDa component and the material found at 80 kDa exerted non-specific immunosuppressive effects. We conclude therefore that isotype-specific IgG-BF has an apparent Mr of 40000.     

Decreased UDP-GlcNAc:Glycopeptideβ-2-N-Acetylglucosaminyltransferase II activity in a ricin-resistant mutant of baby hamster kidney (BHK) cells

The ricin-resistant mutant baby hamster kidney (BHK) cell line RIC^R21 is unable to make the sialylated bi- or triantennary complexN-glycans found in wild type cells and accumulates instead non-bisected hybrid structures containing three Man residues and one or two sialylated antennae (Hugheset al 1983, Carbohydr Res 120215-34). Specific assays forN-acetylglucosaminyltransferases I, II, III and IV were applied to Triton X-100 extracts of wild type BHK, RIC^R14 and RIC^R21 cells. It was shown that RIC^R21 cell extracts had a decreasedN-acetylglucosaminyltransferase II specific activity (17 to 27% of wild type values). It is suggested that in wild type cellsN-acetylglucosaminyltransferase II action proceeds quickly, leading to complexN-glycan synthesis, while in RIC^R21 cells potential substrates forN-acetylglucosaminyltransferase II move into the trans-Golgi compartment before the transferase can act, thereby leading to hybrid structures.     

Cytogenetic study of an interspecific cross of Nicotiana debneyi X N. umbratica

Summary Interspecific F₁ hybrids of Nicotiana debneyi Domin (2n=48) and N. umbratica Burbidge (2n=46), both belonging to the section Suaveolentes, showed a high degree of meiotic chromosome pairing. Two of the five F₂ plants obtained exhibited chromosome mosaicism. The first colchiploid generation (C₁) had the expected chromosome number of 2n=94 while C₂ showed 2n=88, a loss of three pairs of chromosomes. This same chromosome number continued in further colchiploid generations, followed up to C₅, except for a few plants in C₃ which showed chromosome mosaicism. The F₁ phenotype was stable through C₁ to C₅ and fertility was normal in colchiploids through all generations in spite of the loss of three pairs of chromosomes and chromosome mosaicism. This stability and fertility apparently reflect the tolerance of the genomes to the genetic adjustment of chromosome complements which is believed to be associated with the originally polyploid nature of the parental species and the chromosome doubling brought about in the amphidiploids.     

In vitro construction of a recombinant adenovirus Ad2:Ad5

A hybrid virus containing the left half of the Ad5 genome and the right half of the Ad2 genome has been constructed by ligating together in vitro the BamHI.-A fragment of Ad5 (map co-ordinates 0–59.5) to the-SawHI-A fragment of Ad2 (map coordinates 59.5–100), and using this DNA to transfect susceptible cells. Viable progeny virus has been obtained which grows as well as the parental virus without any requirement for helper virus, and probably contains a hybrid hexon polypeptide consisting of the major part of the Ad5 hexon with an Ad2 carboxy terminus.     

Murine leukemia cell hybrids: The quantity of TL antigens expressed by parental and hybrid cells fails to correlate with their sensitivity to TL antibody and complement

The quantity of thymus-leukemia (TL) antigens expressed by murine leukemia cells is significantly greater than that expressed by somatic hybrids of such cells. Based upon the results of ¹²⁵I-lactoperoxidase labeling and antibody absorption procedures, and corrected for size differences between the two cell types, the quantity of TL antigens expressed by RADA-1 cells, a radiation-induced murine leukemia cell line of strain A/J mice, is approximately 5.0 times greater than that of somatic hybrids of RADA-1 and LM(TK)^? cells. LM(TK)^? cells are a thymidine kinase-deficient TL(-) mouse fibroblast cell line. The quantity of TL antigens expressed is related only in part to their susceptibility to lysis by TL antibodies and guinea pig complement (GPC). RADA-1 cells resist lysis. The quantity of TL antigens expressed by RADA-1 cells is analogous to that formed by nonneoplastic thymocytes obtained from F₁ hybrids of two strains of TL(+) and TL(-) mice; cells from both strains are sensitive to TL antiserum and GPC. ASL-1 cells, a spontaneously occurring leukemia cell line of A/J mice, express TL antigens in significantly higher quantities than any of the cell types examined. Exposed to TL antisera, the quantity of TL antigens of ASL-1 cells, but not that of hybrid cells, gradually diminishes. ASL-1 cells convert over a 6-h period of exposure to antibody and guinea pig complement (GPC) resistance; hybrid cells remain sensitive. However, ASL-1 cells converted to TL antibody and GPC resistance continue for a time to express TL antigens in quantities similar to that of sensitive F₁ thymocytes and resistant RADA-1 cells. RADA-1 X LM(TK)^? hybrid cells, which are sensitive to TL antibodies and GPC, express the lowest quantities of TL antigens of any of the cell types examined. It is likely that differences in the quantities of TL antigens expressed by different cell lines reflect genetic mechanisms controlling TL antigen expression. The failure of TL antisera to affect the quantities of TL antigens expressed by hybrid cells is taken as an indication that genetic controls governing antigen expression may be distinguished from those involved in regulating responsiveness to specific antiserum.     

Gene fusion vectors based on the gene for staphylococcal protein A

Two plasmid vectors, containing the gene coding for staphylococcal protein A and adapted for gene fusion, have been constructed. These vectors will allow fusion of any gene to the protein A gene, thus giving hybrid proteins which can be purified, in a one-step procedure, by IgG affinity chromatography. As an example of the practical use of such vectors, the protein A gene has been fused to the lacZ gene of Escherichia coli. E. coli strains containing such plasmids produce hybrid proteins with both IgG binding and β-galactosidase activities. The hybrid protein(s) can be immobilized on IgG-Sepharose by its protein A moiety with high efficiency without losing its enzymatic activity and they can be eluted from the column by competitive elution with pure protein A. The fused protein(s) also binds to IgG-coated microtiter wells which means that the in vivo product can be used as an enzyme conjugate in ELISA tests.     

红金麻——木槿属一个新杂种

<正> 红金麻 新杂种 Hibiscus×hongjinma Y. T. Chang, hybr. nov. H. cannabinus L. ♀× H. radiatus Cav.♂ Hybrida hortensis ( 2n=30+6sat) θ Hibisco cannabino L. ( 2n=32+4sat) et H. ranndiato Cav. (2n=70+2sat) exorta, Habitu illo simil, characteribus ad hoc magis accedit; ab illo differt floribus purpureis, folio prope basin 3—5     

Rat immunoglobulin delta heavy chain gene: nucleotide sequence derived from cloned cDNA

Rat immunoglobulin δ heavy-chain mRNA has been isolated. RNA blot analysis revealed that this mRNA with a length of 1.8 kb encodes for the secreted form of IgD. The corresponding cDNA was cloned in plasmid pBR322 and its sequence was determined. The hybrid plasmid contains a 775-bp insert comprising a partial Cδ₁ sequence and complete CδH, Cδ3, CδDC and 3' untranslated sequences. Rat and mouse IgD amino acid sequences show striking homology in Cδ3 and CδDC regions.     

Toxic ligand conjugates as tools in the study of receptor-ligand interactions

We have constructed hybrid proteins in which the toxic A chains of ricin or diptheria toxin have been linked to either asialofetuin, fetuin, or epidermal growth factor (EGF). Both ASF-RTA and ASF-DTA are potent toxins on cultured rat hepatocytes, cells that display the asialoglycoprotein receptor. Toxicity of these two compounds is restricted to hepatocytes and can be blocked by asialoglycoproteins but not the native glycoproteins or asialoagalactoglycoprotein derivatives, indicating that the toxicity of the conjugates is mediated by the hepatic asialoglycoprotein receptor. The EGF-RTA conjugate is an extremely potent toxin on cells that can bind the hormone, but is only poorly effective on cells that are unable to bind EGF. The EGF-DTA conjugate, in contrast, is unable to kill 3T3 cells and is at least two orders of magnitude less effective than EGF-RTA on A431 cells, a cell line with 1-2 X 10(6) EGF receptors per cell. However, when EGF-RTA and EGF-DTA were tested on primary liver hepatocyte cultures, which were susceptible to both ASF-RTA and ASF-DTA, both EGF conjugates were potent toxins. Sensitivity of the hepatocyte cultures to ricin toxicity increases slightly during a 52-hr culture period. In contrast, sensitivity to EGF-RTA and ASF-RTA decline dramatically during this period. Receptors for both ligands remain plentiful on the cell surface during this time.