Akimitsu Tomonaga

Unexpected Tissue Distribution of Liposomes Coated with Amylopectin Derivatives and Successful Use in the Treatment of Experimental Legionnaires’ Diseases

Bacterial and plant cell membranes are covered by cell walls of polysaccharide derivatives. The cell walls do not only serve to maintain the shape and stiffness of cells and to protect the cell membranes against external stimuli but also play an important role in various biological recognition processes, for instance antigen-antibody interaction, toxin recognition, and cell-cell adhesion,

Sandwich enzyme immunoassay of tumor-associated antigen sialosylated Lewisx using β-d-galactosidase coupled to a monoclonal antibody of IgM isotype

Enzyme conjugates with antibody of IgG type have been used extensively in immunohistochemistry, but conjugates with antibody of IgM type have not been reported. This paper describes the beta-D-galactosidase (Gal) labeling of a monoclonal IgM antibody designated CSLEX1 (for cytotoxic sialosylated Lewisx), which is directed against a tumor-associated antigen sialosylated Lewisx (S-Lex). The antibody was first acylated with a heterobifunctional agent N-(gamma-maleimidobutyryloxy)succinimide (GMBS) to introduce the maleimide groups into the molecule; excess reagent was removed by gel filtration and then the activated antibodies were crosslinked to the thiol groups of Gal. The conjugates were partially purified of free Gal by DEAE-Toyopearl column chromatography with an increasing linear concentration of NaCl. The conjugates thus prepared retained almost full enzyme activity and were demonstrated to be free of CSLEX1 by affinity chromatography using anti-galactosidase antibody bound to Sepharose 4B. The conjugates were used as a label in a sandwich enzyme immunoassay (SEIA) to detect the antigen at concentrations as low as 0.2 U/well. The SEIA was used to measure serum S-Lex levels in both healthy subjects and lung cancer patients and mean concentrations of 70 U/ml and 198.6 U/ml were detected respectively.

Effect of Membrane Fluidizers on the Number and Affinity of Chemotactic Factor Receptors on Human Polymorphonuclear Leukocytes

Chemotaxis by leukocytes appears to be initiated by the binding of chemoattractants to specific cell surface receptors. In other biological systems, the affinity and functional activity of membrane receptors are regulated by the local microviscosity. The present studies were undertaken to determine if the number and/or affinity of chemotactic factor receptors expressed on human polymorphonuclear leukocytes were similarly affected. Aliphatic alcohols and cis‐vaccenic acid, agents known to decrease membrane microviscosity, were studied for their effects on the binding of the radiolabeled chemoattractant f‐Met‐Leu‐[3H]Phe to human polymorphonuclear leukocytes. Butanol and propanol increased the number of f‐Met‐Leu‐[3H]Phe binding sites approximately 1.5 fold. More dramatically, these same agents enhanced the affinity of the receptor by ten‐fold, without affecting the specificity of the receptor. Similarly, cis‐vaccenic acid enhanced both the number and affinity of this chemotactic factor receptor on human polymorphonuclear leukocytes contain cryptic receptors for the N‐formylated peptide chemotactic factors, but more importantly that the affinity of these receptors can exist in more than one state and can be modulated by membrane microviscosity. Alterations of membrane fluidity in leukocytes during chemotaxis may be an important mechanism for regulating their sensitivity to chemoattractants.

Targeting Cancer therapy in Mice by Use of Newly Developed Immunoliposomes Bearing Adriamycin

AbstractPolysaccharide-coated liposomes have been developed to improve the stability of conventional liposomes against biochemical and physicochemical stimuli. Pullulan (MW 5 × 104) was used as the polysaccharide. the mouse IgM monoclonal antibody (CSLEX 1) recognizes a sialosylated Lex, which is a tumor-specific antigen in athymic mice. the IgM antibody was reduced with cysteine to obtain the subunit (IgMs) that remained biologically active. the IgMs was accumulated in an antigen-positive tumor in vivo. Subsequently, it was conjugated with the pullulan-coated liposome to form an immunoliposome.Tissue distribution studies demonstrated that immunoliposomes were more efficiently targeted to an implanted tumor than to the polysaccharide-coated liposomes. This is accompanied by a drastic decrease in liver uptake of the immunoliposomes. Furthermore, adriamycin-encapsulated immunoliposomes in-hibited the growth of the implanted tumor more effectively than did the simple pulluian-coated liposomes.

Production of monoclonal antibodies against Legionella pneumophila serogroup 1

Four monoclonal antibodies to Legionella pneumophila Philadelphia 1 were produced by the fusion of immunized BALB/c lymphocytes to a murine myeloma cell line. Two (Lpl‐1 and Lpl‐3) of the four monoclonal antibodies reacted with 14 L. pneumophila serogroup 1 strains, and the other (Lpl‐2 and Lpl‐4) reacted with only three out of 20 strains tested. These four monoclonal antibodies did not bind to any strains of L. pneumophila serogroup 2–7 and other Legionella species. In addition, it has been shown that these monoclonal antibodies may be useful not only for subserotyping of L. pneumophila but also for retrospective diagnosis using immunopathological methods.

Superoxide production of polymorphonuclear leukocytes in surgical patients with gastric cancer

SummarySuperoxide production (SOP) by polymorphonuclear leukocytes (PMNs) in 65 gastric cancer patients, who were in preoperative state and had received no medical therapy, was assayed in order to evaluate the bactericidal activity of PMNs in cancer patients, as well as to determine the correlation of SOP by PMNs with postoperative prognoses and several factors by which extent of disease and the clinical or histological character of gastric carcinoma were defined. Patients with stage II disease had a tendency to have an increased SOP by PMNs, and furthermore, as the disease progressed, the SOP by PMNs decreased with significant depression being noted in stage III and IV cases compared to healthy controls. Significantly reduced SOP by PMNs was observed in n2 and n3 cases and with se, si, sei and/or ps( + ) pathological invasion. SOP by PMNs in patients with Borrmann II type and/or poorly differentiated adenocarcinoma was significantly depressed. Patients who suffered from septic complications showed a significant depression in SOP by PMNs compared with the controls and no complication group.These results suggest that advanced gastric cancer patients may have defective oxidative PMN metabolism, and that a decrease of SOP is a contributory cause of high susceptibility to postoperative infection in cancer patients.

A case of benign asbestos pleurisy as pleural bloody effusion

50才男性でアスベストにより血性胸水を生じた1例である.症例は30年間のアスベスト暴露歴を有し,昭和58年12月胸部写真で胸水貯留を無症候性に認め,昭和59年2月胸水が増加して入院精査した.胸水は血性滲出液で細胞診や細菌培養は陰性.喀痰中や気管支肺胞洗浄液中に多数のアスベスト小体が証明され,悪性腫瘍や結核などを否定し,無治療で経過観察をした. 6カ月後に胸水が減少し,胸膜肥厚や肺の線維化が認められ, 2年6カ月後の現在も悪性腫瘍の発生はなく,アスベストによる良性胸水貯留と考えられた.アスベストによる良性胸水貯留と診断しえた症例の報告は,本邦において少なく,その診断に気管支肺胞洗浄が有用と思われた.

A case of primary lung cancer with diffuse sarcoid granuloma in the resected lung.

症例は74才の女性. 7年前よりブドウ膜炎および緑内障にて治療を受け, 胸部X線にて右上肺野に腫瘤影を指摘されていたが放置していたところ, 同陰影が次第に増大の傾向にあるため精査し, 肺腺癌の診断にて右上葉切除を受けた. 切除肺には癌部および非癌部ともにサルコイド肉芽腫をびまん性に認め, 術後の精査にて血清ACE値や, 対側肺の気管支肺胞洗浄液などから, サルコイドーシスの合併が疑われた症例を経験したので報告した.

Clinical evaluation of neutrophil polarization in surgical patients with gastric cancer.

胃癌患者60例の末梢血好中球polarizationを測定し, 胃癌の病態因子別に検討を加えた.胃癌患者の好中球polarizationは対照群に比べ有意に低下していた.stage IV, n3, 深達度ssで示される進行胃癌患者で有意な低下を認めた.術後感染症合併例では, 他群に比較して差を認めなかった.死亡例では有意な低下をみた.C領域癌, Borrmann 3型癌, por (低分化腺癌) 癌で有意な低下を示した.これらの成績から進行胃癌患者では, 一連の好中球機能の過程で最も初期に位置するpolarizationという機能においてその低下が示され, 担癌患者での好中球機能不全が示唆された.