Relationships of the soluble human A, B, and H antigens of blood group A1B individuals

Ouchterlony double diffusion reactions with precipitating antibodies and lectins provide visual evidence that in saliva of A1B secretor individuals the A and B antigens are predominantly situated on the same molecule, while the H antigen is on a different molecule.     

Prianicin A and B,nor-sesterterpenoid peroxide antibiotics from red sea sponges

Summary The antibiotic properties of 2 acidic C₂₄H₄₀O₄ nor-sesterterpenoid peroxides, prianicin A (1) and B (2), against gram-positive and gram-negative bacteria, and against fungi are herein described. They are 4–10 times more effective than tetracycline againstbeta hemolytic Streptococcus, but significantly non-effective against a variety of gram-negative bacteria.Acknowledgment. The authors are grateful to Prof. A. Kjaer of the Organic Chemistry Department of the Technical University of Denmark, for his help and fruitful discussions.     

Cannflavin A and B,prenylated flavones fromCannabis sativa L.

Summary Two novel prenylat flavones, termed Cannflavin A and B, were isolated from the cannabinoid free ethanolic extract ofCannabis sativa L. Both compounds inhibited prostaglandin E₂ production by human rheumatoid synovial cells in culture.This communication is dedicated to the memory of the late Professor J. W. Fairbairn and the late Dr. J. T. Pickens. We are grateful to the Medical Research Council for a project grant and to Ms J. Elliot of King's College London for NMR spectra.     

Vidalols A and B,new anti-inflammatory bromophenols from the Caribbean marine red algaVidalia obtusaloba

Chemical studies of the Caribbean red algaVidalia obtusaloba have resulted in the isolation of two new bromophenolic metabolites, vidalols A and B (1, 2). The new compounds were discovered as part of an organized effort to isolate new naturally-occurring anti-inflammatory agents with a focus upon those which may function through the inhibition of phospholipase A₂.     

Isolation of the two paragonial substances deposited into the spermatophores ofAcanthoscelides obtectus (Coleoptera,Bruchidae)

Summary Aqueous extracts from spermatophores ofAcanthoscelides obtectus (Coleoptera, Bruchidae) have been chromatographed on Sephadex G 25 and G 15 columns, the active fractions being further purified by preparative SiO₂ TLC. The isolation and properties of 2 biologically active compounds are reported. The paragonial substance A has a favourable action upon oogenesis, is a low molecular weight molecule (500–1000) and gives 1 single amino acid on hydrolysis, corresponding to glycin on TLC. The paragonial substance B is toxic, or an inhibitor of oogenesis at lower concentration, thus showing an antagonistic effect.     

Oxidation of synephrine by type A and type B monoamine oxidase

Summary Synephrine (SP) was found to be a substrate for monoamine oxidase (MAO) in rat brain mitochondria, showing the K_m and V_max values of 250 M and 32.6 nmoles/mg of protein/30 min respectively. The inhibition studies showed that the SP oxidation was carried out by both type A and type B MAO and a major part of the activity was due to type A MAO.     

Large-scale purification of hepatitis B surface antigen using affinity chromatography

Summary Large-scale purification of hepatitis B surface antigen, applicable to the preparation of potential vaccines for prevention of hepatitis B, is described. The method involves the following steps: precipitation of serum with polyethylene glycol 6000, affinity chromatography on concanavalin A-Sepharose and on -aminononyl-Sepharose, and isopycnic centrifugation.This study was supported by grant 09011 from the National Heart, Lung and Blood Institute. We thank Dr. B. Hollinger for testing HB_sAg samples by a complement fixation test.     

Augmentation of natural antiganglioside IgM antibodies in lower motor neuron disease (LMND) and role of CD5+ B cells

IgM antibodies directed against neuronal gangliosides GM₁ , GM₂ , GD_1a , GD_1b and GT_1b occur in normal individuals and their level significantly decreases with age. Patients with lower motor neuron disease (LMND) produce high levels of these autoantibodies. AntiGM₁ IgM is selectively augmented. In these patients, the CD5+ (B1) and CD5− (B2) subsets of B cells are not distinct entities but range from those without detectable CD5 marker to those with high CD5+ expression. B1 B cells were sorted to homogeneity, but B2 B cell cannot be isolated to homogeneity because of the presence of B1 cells with low CD5 expression. In short term cultures both the subsets produced IgM antibodies, but the antibodies reacted better with desialylated GM₁ than with GM₁ . Cycloheximide (Cx) (0.35 mM) largely blocked IgM synthesis of the B1 B cells but inhibition of the B2 B cells was incomplete, possibly due to shedding of cytophilic antibodies as well as to the presence of B1 phenotype with loss of CD5 expression. CD5+ B cells may be involved in the production of antiglycolipid IgM. Received 9 June 1997; received after revision 21 July 1997; accepted 28 July 1997     

Adsorption of hepatitis B surface antigen to matrix-bound long chain hydrocarbon structures

Summary The binding of hepatitis B surface antigen (HB_sAg) to various matrix bound long-chain hydrocarbon structures has been studied. It was found that HB_sAg was strongly bound to straight hydrocarbon chains with more than seven carbon atoms. The adsorbents can probably be used for removal and /or detection of hepatitis B infectious material.     

Cloning of the hepatitis B virus genome in Escherichia coli]

The whole genome of the hepatitis B virus (Dane particles) was inserted in vitro in the genome of the bacteriophage lambda gtWES . LAMBDA B. The recombinant DNA molecule was cloned in E. coli. Amplification of the hybrid bacteriophage enables the preparation of large amounts of hepatitis B virus DNA. The possibilities offered by the utilization of this recombinant bacteriophage are discussed.     

Localization of human A,B and H isoantigens in Cynomolgus monkey tissues

Summary The presence and distribution of human A, B and H isoantigens were demonstrated in Cynomolgus monkey (Macaca fascicularis) by means of red cell adherence test. Although no human antigens were found on primate erythrocytes, various epithelial tissues revealed the presence of A, B or H antigenic substance. The distribution and localization was similar to that found in human tissues. Majority of specimens from each individual animal possessed only 1 human type isoantigen with the exception of the salivary and sweat glands, where all animals showed the presence of H antigen in addition to other specificity, and of Brunner's gland, where all sections reacted positively also for A antigen.     

Aufnahme von Ferrioxamin B durch Tomatenpflanzen

Summary Ferrioxamin B (FOB), an iron chelating cyclopeptide, is shown to be absorbed and translocated in tomato plants from nutrient solution. Chelated iron (FOB) is transported to the upper parts of tomato plants more rapidly than ionic iron (FeCl₃).     

Presence of the H blood group antigen on a carcinoembryonic antigen (CEA). Enzymatic transformation of H to A and B specificities]

The carcinoembryonic antigen (CEA-M) was purified from a hepatic metastasis obtained from a blood group O patient with a cancer of the rectum. Using 125I-labelled-CEA and blood group antisera, H specificity was found on the CEA-M; the addition of anti-H to anti-CEA does not modify the binding of labelled-CEA-M to its antibodies (86%), this result leads us to conclude that H and CEA determinants are carried by the same molecule. However the low percentage of binding (30% with 1/10 anti-H) suggests that only a few CEA-M molecules do carry the H antigenic determinant. Finally, glycosyltransferases were used to modify the H specificity into blood group A and B specificities.     

The isolation and structure of trichosiderin B

Riassunto Un nuovo pigmento feomelanico, denominato tricosiderina B, è stato isolato dalle piume di pollo di razza New Hampshire. Sulla base delle proprietà chimiche e spettrali a tale pigmento, C₂₃H₂₀N₄O₉S₂, è stata assegnata la struttura ^2,2-bibenzotiazinica IV.

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This work was supported by a grant from Laboratorio per la Chimica e Fisica di Molecole di Interesse Biologico del C.N.R., Napoli.     

The isolation and structure of trichosiderin B

Riassunto Un nuovo pigmento feomelanico, denominato tricosiderina B, è stato isolato dalle piume di pollo di razza New Hampshire. Sulla base delle proprietà chimiche e spettrali a tale pigmento, C₂₃H₂₀N₄O₉S₂, è stata assegnata la struttura ^2,2-bibenzotiazinica IV.

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This work was supported by a grant from Laboratorio per la Chimica e Fisica di Molecole di Interesse Biologico del C.N.R., Napoli.     

Semisynthesis and biological properties of the [B24-leucine]-, [B25-leucine]- and [B24-leucine,B25-leucine]-analogues of human insulin

Summary Trypsin-catalyzed coupling of porcine desoctapeptide-insulin with synthetic octapeptides produced the [Leu^B24]-(I), [Leu^B25]- (II) and [Leu^B24, Leu^B25]- (III)analogues of human insulin. I, II and III displayed respectively 20–30%, 1–2% and 0.5% of the receptor binding activity of the normal hormone. Biological activities of these analogues seemed to be proportional to their binding potencies when assayed in vitro, while in an in vivo assay analogue I was fully active and II exhibited 10–20% of normal activity. III was less active than II in all assays tested.     

Efficacy of tumor cell vaccine after incorporating monophosphoryl lipid A (MPL) in tumor cell membranes containing tumor-associated ganglioside

Murine B16 melanoma expresses the ganglioside. GM₃. GM₃ shed from tumor cells is immunosuppressive and promotes tumor growth¹. Reduction or elimination of the shed GM₃ could be therapeutic, and the anti-GM₃ antibodies may reduce and clear the shed ganglioside. To test this hypothesis, mice were challenged with tumor cells, with or without inducing anti-GM₃ antibody response. Since gangliosides are poor immunogens and T-cell independent antigens, an adjuvant (monophosphoryl lipid A (MPL), a non-toxic lipid A ofSalmonella), directed against B-cells, was employed. MPL was incorporated onto liposomes and into the surface membrane of B16 mouse melanoma cells; both are rich in GM₃. C57BL/6J mice immunized with MPL-liposomes or MPL-B16 cells responded with elevated levels of anti-GM₃ IgM. Non-immunized mice or mice immunized with B16 cells alone or ganglioside GM₃ alone (without MPL) elicited poor anti-GM₃ IgM response, confirming the GM₃'s immunologic crypticity and MPL's immunopotentiating effect. MPL's immunopotentiating effect was improved by coupling it to melanoma cell membranes C57BL/6J mice were immunized with irradiated B16 alone or MPL alone or MPL-conjugated irradiated B16. After three weekly immunizations, each mouse received a challenge dose of viable syngeneic B16. Neither MPL alone nor B16 alone had a significant effect on tumor growth or host survival; however, administration of MPL-conjugated B16 cells significantly prevented tumor growth and prolonged survival. Our results indicate that MPL-incorporated B16 cells augment the anti-GM₃ IgM response, which may reverse GM₃-induced immunosuppression by eliminating tumor-derived GM₃, and restore immunocompetence.     

Multimeric structures of HLA-G isoforms function through differential binding to LILRB receptors

The non-classical Human leukocyte antigen G (HLA-G) differs from classical HLA class I molecules by its low genetic diversity, a tissue-restricted expression, the existence of seven isoforms, and immuno-inhibitory functions. Most of the known functions of HLA-G concern the membrane-bound HLA-G1 and soluble HLA-G5 isoforms, which present the typical structure of classical HLA class I molecule: a heavy chain of three globular domains α₁–α₂–α₃ non-covalently bound to β-2-microglobulin (B2M) and a peptide. Very little is known of the structural features and functions of other HLA-G isoforms or structural conformations other than B2M-associated HLA-G1 and HLA-G5. In the present work, we studied the capability of all isoforms to form homomultimers, and investigated whether they could bind to, and function through, the known HLA-G receptors LILRB1 and LILRB2. We report that all HLA-G isoforms may form homodimers, demonstrating for the first time the existence of HLA-G4 dimers. We also report that the HLA-G α₁–α₃ structure, which constitutes the extracellular part of HLA-G2 and HLA-G6, binds the LILRB2 receptor but not LILRB1. This is the first report of a receptor for a truncated HLA-G isoform. Following up on this finding, we show that the α₁–α₃-Fc structure coated on agarose beads is tolerogenic and capable of prolonging the survival of skin allografts in B6-mice and in a LILRB2-transgenic mouse model. This study is the first proof of concept that truncated HLA-G isoforms could be used as therapeutic agents.     

New antibiotics,trichopolyns A and B: Isolation and biological activity

Summary Polypeptide antibiotics, trichopolyns A and B, were isolated from the culture broth ofTrichoderma polysporum (Link ex Pers) Rifai (TMI 60146). Assessment of biological activity of the antibiotics against microorganisms was made.Acknowledgment. We are most grateful to Central Research Division, Takeda Chemical Industry Co. Limited, for determination of antifungal and antibacterial spectra.     

Lysosomal chymotrypsin B potentiates apoptosis via cleavage of Bid

We have reported that chymotrypsin B (CtrB) is not just a digestive enzyme but is also stored in lysosomes. Herein, we demonstrated a broad distribution of CtrB and explored the involvement of CtrB in apoptosis. Exposure of RH-35 cells to H₂O₂ or palmitate induced the redistribution of lysosomal CtrB into the cytoplasm as a result of lysosomal membrane permeabilization (LMP). Suppression of CtrB significantly blocked apoptosis, while overexpression of CtrB sensitized apoptosis markedly. CtrB could cleave Bid under neutral conditions. In RH-35 cells with Bid silenced, apoptosis induced by CtrB protein was attenuated, suggesting that CtrB mediates apoptosis of RH-35 cells mainly through processing Bid. Our data also suggest that LMP occurs earlier than mitochondrial outer membrane permeabilization; Bid activation initiated by caspase-8 might be reinforced by CtrB in consequence of LMP, which causes a positive feedback loop leading to the accumulation of tBid, and results in lysosome- and mitochondrion-dependent apoptosis.