A human monoclonal antibody directed to blood group i antigen: heterohybridoma between human lymphocytes from regional lymph nodes of a lung cancer patient and mouse myeloma

A fusion of human lymphocytes released from regional lymph nodes of papillary adenocarcinoma of lung cancer with mouse myeloma P3-X63-Ag8-U1 cells resulted in a stable hybridoma-secreting human IgM antibody (NCC-1004) that reacts with a large proportion of squamous cell carcinomas of lung and esophagus as well as carcinoma of thyroid glands. However, the antibody also reacts with normal red blood cells, B lymphocytes, and a few other limited loci in normal tissues such as the basal cells of bronchial epithelium and the basal cell layer of stratified squamous epithelium, as well as endothelium and alveolar lining epithelium. The antigen defined by NCC-1004 has been characterized as blood group i antigen on the basis of the following results. The antibody preferentially agglutinates cord erythrocytes in contrast to adult erythrocytes. The agglutination was obvious at 4 degrees C, but diminished greatly at 37 degrees C, and was enhanced after sialidase treatment. The antibody specifically reacts with lacto-norhexaosylceramide (nLc6) and sialosyllacto-norhexaosylceramide (IV3NeuAcnLc6), but does not react with lacto-neotetraosylceramide (nLc4), sialosyllacto-neotetraosylceramide (IV3NeuAcnLc4), lacto-isooctaosylceramide (IV6Gal beta 1----4GlcNAcnLc6; I antigen), and other standard glycolipids so far tested. The properties of the antibody and its antigen are identical to those previously described for the i blood group system. Inasmuch as the hybridoma was established by hybridization of lymphocytes derived from regional lymph nodes of lung cancer, and the antigen was found in the patient's lung cancer tissue, the i antigen in lung cancer is probably recognized as a tumor-associated antigen by the host's immune cell system.     

Activated c-raf gene in a rat hepatocellular carcinoma induced by 2-amino-3-methylimidazo[4,5-f]quinoline

A rat hepatocellular carcinoma, IQ7, induced by 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) gave two transformants of NIH 3T3 cells on DNA mediated gene transfer. One of these transformants was examined further and secondary and tertiary transformants were obtained. The secondary transformant was tumorigenic in nude mice. The activated oncogene in this primary transformant was identified as rat c-raf by Southern blot analysis.     

Thermostable tryptophan synthase ofBacillus stearothermophilus expressed inEscherichia coli

Summary The tryptophan synthase genes,trpA andtrpB, from a moderate thermophile,Bacillus stearothermophilus IFO13737, were expressed efficiently inEscherichia coli. The recombinant tryptophan synthase amounted to 22% of the soluble cellular protein, and was purified to homogeneity by three steps. The enzyme is more thermostable thanE.coli tryptophan synthase, especially the subunit. The enzyme is also more resistant to sodium dodecylsulfate and methanol thanE.coli enzyme.     

Assignment of the CD45-AP Gene to the Centromeric End of Mouse Chromosome 19 and Human Chromosome 11q13.1–q13.3

CD45-AP is a recently identified phosphorylated protein that specifically associates with the leukocyte-specific transmembrane glycoprotein CD45. The gene for CD45-AP,Ptprcap(protein tyrosine phosphatase, receptor type c polypeptide associated protein), was mapped in mouse by typing the progeny of two multilocus crosses using the mouse CD45-AP cDNA as a Southern hybridization probe. The CD45-AP gene mapped to the centromeric region of Chr 19 proximal to the genesFth, Cd5,andPcna-rs.The gene for the human CD45-AP homologue,PTPRCAP,was localized to chromosome band 11q13.1–q13.3 by fluorescencein situhybridization using human genomic CD45-AP DNA as a hybridization probe. The genetic mapping of thePtprcap/PTPRCAPgenes extends the previously defined synteny conservation of various genes that have been assigned to these regions of the mouse and the human chromosomes.     

Two new subspecies of Bacillus thuringiensis isolated in Japan: Bacillus thuringiensis subsp. kumamotoensis (serotype 18) and Bacillus thuringiensis subsp. tochigiensis (serotype 19)

Bacteriological and serological characteristics of three Bacillus thuringiensis isolates obtained in Japan were investigated. They formed typical rhomboidal parasporal inclusions but flagellar (H) antigens of these isolates were different from those of the known 17 H serotypes of B. thuringiensis. The three isolates were divided into two new serotypes (serotypes 18 and 19). The serotype 18 isolate (3–71) produced thermostable exotoxin and the inclusions of this isolate were toxic to larvae of the silkworm, Bombyx mori, but nontoxic to larvae of the mosquito, Aedes aegypti. The other isolate (119-72) belonging to serotype 18 produced inclusions nontoxic to larvae of B. mori and A. aegypti and did not produce thermostable exotoxin. However, other bacteriological properties of the isolate 119-72 were similar to those of the isolate 3–71. The serotype 19 isolate (117-72) produced inclusions nontoxic to larvae of B. mori and A. aegypti and did not produce thermostable exotoxin. Acid production from saccharose and the production of brownish purple pigment were observed in the two serotype 18 isolates, while neither of them was observed in the serotype 19 isolate. In other 29 biochemical properties tested, there was no difference among the three isolates. Based on these characteristics, the following two subspecies names are proposed: Bacillus thuringiensis subsp. kumamotoensis (serotype 18) for the type strain 3–71 and Bacillus thuringiensis subsp. tochigiensis (serotype 19) for the type strain 117-72.     

Synthesis of D-manno-3-heptulose,D-ido-3-heptulose,and some aldoheptoses via isopropylidene derivatives of heptitols

D-manno-3-Heptulose (5) was synthesized by dimethyl sulfoxide-phosphorus pentaoxide oxidation of 1,2:3,4:6,7-tri-O-isopropylidene-D-glycero-D-manno-heptitol (3, prepared from volemitol), followed by hydrolysis. D-ido-3-Heptulose (8) was synthesized similarly by oxidation of 1,2:4,5:6,7-tri-O-isopropylidene-D-glycero-l-galacto-heptitol (7, prepared from D-glycero-l-galacto-heptitol, 6). Another tri-O-isopropylidene derivative (11), having a free primary hydroxyl group, was produced in larger amount than 7, and 11 yielded D-glycero-l-galacto-heptose (14). Compound 8 was also synthesized by way of 1,2:4,5.6,7-tri-O-isopropylidene-D-glycero-l-gulo-heptitol (15). The production of 15 from D-glycero-l-gulo-heptitol (13) was accompanied by a larger amount of 2,3:4,5:6,7-tri-O-isopropylidene-D-glycero-D-ido-heptitol (17) which, upon oxidation followed by hydrolysis, yielded D-glycero-D-ido-heptose (18). One of the two tri-O-isopropylidene derivatives obtained by acetonation of perseitol, 2,3:4,5:6,7-tri-O-isopropylidene-D-glycero-D-galacto-heptitol (19), yielded D-glycero-D-galacto-heptose (20).     

Substance P radioimmunoassay using Nalpha-tyrosyl-substance P and demonstration of the presence of substance P-like immunoreactivities in human blood and porcine tissue extracts.

Sensitive and specific radioimmunoassay for substance P was developed using synthetic substance P and 125I-Nalpha-tyrosyl-substance P. Substance P-human alpha-globulin conjugate was used for production of anti-substance P antisera in rabbits. Synthetic substance P was used as a standard and the dextran-coated charcoal method was employed to separate the free peptide from that bound to antibodies. No cross-reactions by physalaemin and eledoisin observed in this system proved its high specificity to substance P. Nalpha-Tyrosyl-substance P and [Tyr1]-substance P showed the displacement curves indistinguishable from that of the standard substance P. Neither substance P5-11 nor substance P6-11 competed with the tracer at the concentration used. The minimum measurable dose of substance P by the assay system was 2.5-5 pg/incubate. Utilizing the system, human plasma samples from 42 healthy volunteers of both sexes were shown to contain immunoreactive substance P in amounts that averaged 298 pg/ml in male and 251 pg/ml in female. Substance P-like immunoreactivity was also demonstrated in hot-water extracts of porcine duodenum, jejunum, ileum, cecum, middle colon, rectum, pancreas, stomach and pituitary. The highest concentration (379 ng/g wet weight of organ) was found in the pituitary, and the ileum (7.9 ng/g wet weight of organ) and jejunum (1.9 ng/g wet weight of organ) were rich in the contents.