論文 - 丸山 治彦
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Anti-idiotype cancer vaccines: Past and future
Herlyn D., Somasundaram R., Li W., Maruyama H.
Cancer Immunology Immunotherapy 43 ( 2 ) 65 - 76 1996年11月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Cancer Immunology Immunotherapy
Anti-idiotypic antibodies (Ab2) binding to the antigen-combining site of antitumor antibodies (Ab1) can induce anti-anti-idiotypic antibodies (Ab3) that specifically bind to the tumor antigen recognized by Ab1. Furthermore, Ab2, mimicking tumor antigens, have been shown to induce anti-anti-idiotypic proliferative T lymphocytes of the helper and suppressor type, as well as cytotoxic lymphocytes. The immunomodulatory activities of Ab2 have been demonstrated both In animals and in patients. The demonstration of tumor growth inhibition by anti-idiotypes in preclinical and phase I clinical studies emphasizes that randomized control trials should be performed to demonstrate clinical efficacy of Ab2 vaccines.
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Owhashi M., Maruyama H., Nawa Y.
International Journal for Parasitology 26 ( 7 ) 705 - 711 1996年7月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：International Journal for Parasitology
Kinetic changes of eosinophil chemotactic factor (ECF) production from granulomas, splenic T-cells or mast cells were examined with reference to granuloma formation around newly deposited single eggs in Schistosoma japonicum-infected mice. The peri-ovular granulomas began to appear at around 5 weeks post-infection (p.i.). Their size reached a peak at 6 weeks and then decreased gradually. Up to 8 weeks p.i., eosinophils were the predominant cell type in the granulomas. ECF-release from isolated granulomas paralleled the size of granulomas. Circulating ECF-A, which was assumed to be derived from mast cells, was also detected 6 weeks afterwards in parallel with the level of specific IgE antibody level against egg antigens in the serum. The circulating ECF-A peaked at 8 Necks and decreased after 10 weeks. Spleen cells began to produce ECF specific to bone-marrow eosinophils began at 5 weeks p.i., reached a peak at 6 weeks and then decreased rapidly. On the other hand, the production of ECF specific to eosinophils obtained from the peritoneal cavity began at 6 weeks and decreased rapidly thereafter. These results suggest that various kinds of host-derived ECFs seem to contribute, in one way or an other, to tile accumulation of eosinophils in and around granulomatous lesions. The possible role of these ECFs in eosinophil mobilization from the site of production to the inflamed site is discussed.
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Emerging Problems of Parasitic Diseases in Southern Kyusyu, Japan
丸山 治彦, 野田 伸一, 名和 行文
寄生虫学雑誌 = Japanese journal of parasitology 45 ( 3 ) 192 - 200 1996年6月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：日本寄生虫学会
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Fascioliasis Cases Recently Found in the Southern Part of Kyushu District, Japan
丸山 治彦, 野田 伸一, 三森 龍之, 名和 行文
寄生虫学雑誌 = Japanese journal of parasitology 45 ( 3 ) 247 - 254 1996年6月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：日本寄生虫学会
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Itoh H., Murakumo Y., Tomita M., Ide H., Kobayashi T., Maruyama H., Horii Y., Nawa Y.
Biochemical Journal 314 ( 3 ) 923 - 929 1996年3月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Biochemical Journal
By using the combination of reverse-transcription PCR and rapid amplification of cDNA ends methods, two distinct cDNAs encoding mast-cell proteases (chymases; MCPs), designated as gMCP-1 and -2, were successfully cloned and sequenced from the jejunum of Mongolian gerbil, Meriones unguiculatus, infected with Nippostrongylus brasiliensis. On the basis of a comparison of the deduced amino acid sequences with those of known rodent mast-cell chymases, gMCP-1 was found to be highly similar to mouse mast-cell protease (mMCP)-4 and rat mast-cell protease (rMCP)-1, while gMCP-2 was similar to mMCP-5 and rMCP-3: Although mMCP-4 and -5 and rMCP-1 and -3 were restrictedly or mainly expressed in connective-tissue mast cells and serosal mast cells, the gMCP-1 and -2 genes were mainly transcribed in the jejunal mucosa and to a lesser extent in the skin and tongue. Moreover, kinetic study after infection revealed that the amounts of the gMCP-1 and -2 mRNAs in jejunum paralleled well the degree of intestinal mastocytosis. The expression of gMCP-1 and -2 in mucosal mast cells of gerbil jejunum was also confirmed by in situ hybridization. Since a tryptase, another type of MCP, was also expressed in mucosal mast cells of gerbils but not in those of mice and rats, the expression of MCPs in mucosal mast cells of gerbils is different from those of mice and rats. The Mongolian gerbil would be a useful model with which to investigate the physiopathological role of MCPs.
DOI： 10.1042/bj3140923
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Maruyama H., Benden A., Weiping L., Zaloudik J., Koido T., Taupin J., Acres B., Somasundaram R., Prewett M., Herlyn D.
International Journal of Cancer 65 ( 4 ) 547 - 553 1996年2月
担当区分：筆頭著者 記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：International Journal of Cancer
Anti-idiotypic antibodies (Ab2) that bind to the antigen-combining region of anti-tumor antibodies (Ab1) may functionally, and even structurally, mimic tumor antigen. We have previously demonstrated that polyclonal goat Ab2 directed against anti-human gastrointestinal carcinoma Ab1 GA733 induces anti-anti-idiotypic antibodies (Ab3) in animals that are Ab1-like in their binding specificity and idiotope expression. To obtain more defined Ab2 vaccines with potentially increased specificity and efficacy, a monoclonal Ab2 (FG1) was produced against Ab1 GA733 in rats. The monoclonal Ab2 FG1, similar to the polyclonal Ab2 described previously, induced Ab3 in rabbits that were Ab1-like in their idiotope expression and binding specificity to tumor cells and antigen. Antigen-specific Ab3 induced by Ab2 FG1 were easily detected in unprocessed rabbit sera, whereas the demonstration of such Ab3 after polyclonal Ab2 immunization required purification of the Ab3 from the rabbit sera. In addition, Ab2 FG1 induced antigen specific humoral and cellular immunity in mice. Murine Ab3 bound specifically to antigen-positive tumor cells. Ab2-immunized mice showed antigen-specific delayed-type hypersensitivity (DTH) reaction, and cultured splenocytes from the immune mice demonstrated specific proliferation and cytokine (interferon-γ and interleukin-4) secretion upon stimulation with GA733 antigen. However, immune mice were not protected against a challenge with syngeneic GA733 antigen-expressing colon carcinoma cells.
DOI： 10.1002/(SICI)1097-0215(19960208)65:4<547::AID-IJC25>3.0.CO;2-5
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Eosinophil peroxidase deficiency in new zealand white mice
Ohmori J., Tokunaga H., Ezaki T., Maruyama H., Nawa Y.
International Archives of Allergy and Immunology 111 ( 1 ) 30 - 35 1996年1月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：International Archives of Allergy and Immunology
Eosinophil peroxidase (EPO) is one of the granule enzymes in the eosinophil-specific granules and is distinct from myeloperoxidase. Here we report that peroxidase activity was absent in eosinophils of New Zealand White (NZW) mice. When NZW, New Zealand Black and their F, mice were treated with cyclophosphamide followed by Toxocara canis infection, the kinetic changes in the number of eosinophils in peripheral blood, determined by counting in Hinkelman’s diluting fluid, were almost comparable among the three strains. However, when their blood films were stained for peroxidase reaction, eosinophils of NZW mice, but not of the other strains, lacked EPO activity, though their specific granules were stained by eosin Y Sudan black staining for phospholipid was also negative in eosinophils of NZW mice. EPO deficiency in NZW eosinophils was further confirmed by electron-microscopic observations and by measuring EPO activity in the extracts of eosinophil-rich cell suspensions. These results indicate that NZW eosinophils share most of the features with human EPO-deficient eosinophils, suggesting that the NZW mouse is a murine counterpart of human EPO deficiency. © 1996 S. Karger AG, Basel.
DOI： 10.1159/000237341
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Expression of inter-α-trypsin inhibitor light chain (bikunin) in human pancreas
Itoh H., Tomita M., Kobayashi T., Uchino H., Maruyama H., Nawa Y.
Journal of Biochemistry 120 ( 2 ) 271 - 275 1996年
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Biochemistry
Expression of inter-α-trypsin inhibitor light chain (ITI-LC, also known as bikunin or urinary trypsin inhibitor) was examined in various human tissues. By reverse-transcription polymerase chain reaction, the mRNA was detected not only in the liver, a known site of ITI-LC production, but also in the kidney, heart, lung, and pancreas. By RNA blot analysis, the mRNA was also detected in the pancreas and liver, but not in the kidney, heart, or lung. The ITI-LC protein was immunohistochemically detected along the surface of pancreatic acinar cells. These results indicate the apparent expression of the gene for ITI-LC in the pancreas. ITI-LC protein on the surface of pancreatic acinar cells may play an important role in preventing autodigestion by exocrine enzymes such as trypsinogen and chymotrypsinogen.
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Cdna sequencing and expression of rat mast cell tryptase
Ide H., Itoh H., Tomita M., Murakumo Y., Kobayashi T., Maruyama H., Osada Y., Nawa Y.
Journal of Biochemistry 118 ( 1 ) 210 - 215 1995年7月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Biochemistry
A cDNA encoding rat mast cell tryptase (rMCT) was successfully cloned, and sequenced, from peritoneal cells of Lewis rats infected with Nippostrongylus brasiliensis by the reverse transcription-polymerase chain reaction and repid amplification of cDNA ends methods. The cDNA was 1, 097 base-pairs long, and included 822 base-pairs of an open reading frame. As judged from the deduced amino acid sequence, rMCT is highly homologous to mouse mast cell protease-6, and is considered to be translated as a prepro-enzyme with a 19 ami no acid signal peptide, a 10-ami no acid activation peptide, and a 245-amino acid mature enzyme. The rMCT mRNA was not detected in peritoneal cells of mast cell-deficient Ws/Ws rats, though it was strongly detected in oncs of littermate +/+ and Lewis rats. In addition to in peritoneal mast cells, the rMCT mRNA was detected in the tongue. However, mRNA signals were not detected in the small intestine regardless of N. brasiliensis infection. Nor were mRNA signals detected in RBL2H3 rat basophilic leukemia cells. In the lung, the rMCT mRNA was strongly detected after infection with N. brasiliensis though it was only faintly detected before infection. These results suggest that the rMCT is basically specific for connective tissue mast cells, but not for mucosal mest cells and that it is up-regulated in the lung during the inflummatory process of a parasitic infection. © 1995 by the Journal of Biochemistry.
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SOMASUNDARAM R., JACOB L., ADACHI K., CLASS R., SCHECK S., MARUYAMA H., HERLYN D.
Scandinavian Journal of Immunology 41 ( 4 ) 384 - 390 1995年4月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Scandinavian Journal of Immunology
Mice lacking functional T and B lymphocytes offer an in vivo animal model for the study of human immune functions. We have attempted to optimize the reconstitution of severe combined immunodeficiency (SCID) mice with human peripheral blood lymphocytes (PBL) using radiation, anti‐asialo GM 1 antibody or cyclophosphamide (Cy) treatment of the mice and in vitro stimulation of human PBL with interleukin (IL)‐2 prior to their transfer to the mice. Total human IgG and tetanus‐toxoid (TT)‐specific human IgG responses of the mice were used as parameters of successful reconstitution. Treatment of the mice with anti‐asialo GM 1 antibody significantly enhanced total human IgG levels, but not TT‐specific antibody responses, whereas irradiation or Cy treatment of the mice had no effect on human antibody production. In vitro treatment of human PBL with IL‐2 prior to engraftment significantly decreased total human IgG responses of human PBL‐grafted SCID mice. The immune responses of individual mice within a group were highly variable, which constitutes a major disadvantage of this model. Copyright © 1995, Wiley Blackwell. All rights reserved
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Murakumo Y., Ide H., Itoh H., Tomita M., Kobayashi T., Maruyama H., Horii Y., Nawa Y.
Biochemical Journal 309 ( 3 ) 921 - 926 1995年
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Biochemical Journal
By using the combination of reverse-transcription PCR and rapid amplification of cDNA ends methods, a cDNA encoding mast cell tryptase was successfully cloned from the small intestine of Mongolian gerbil, Meriones unguiculatus, infected with Nippostrongylus brasiliensis. The cDNA was 1219 bp long including 810 bp of an open reading frame, Based on the deduced amino acid sequences of known mast cell tryptases of other species, the gerbil mast cell tryptase (gMCT) was highly similar to mouse mast cell protease (mMCP)-7, and seems to be translated as a prepro-enzyme with 25 amino acids of signal and activation peptides and 245 amino acids of mature enzyme. The gMCT mRNA was preferentially transcribed in the intestinal mucosa and to a far lesser extent in the connective tissue such as skin and tongue. Moreover, kinetic study after infection revealed that the amount of gMCT mRNA in the small intestine correlated well with the degree of intestinal mastocytosis. Throughout the course of infection, enzyme-histochemically detectable tryptase activity was limited to mucosal mast cells. Since mucosal mast cells of other rodents, including mice and rats, do not express tryptases, this is the first report of rodent mast cell tryptase expressed in the intestinal mucosa.
DOI： 10.1042/bj3090921
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Ide H., Itoh H., Tomita M., Murakumo Y., Kobayashi T., Maruyama H., Osada Y., Nawa Y.
Biochemical Journal 311 ( 2 ) 675 - 680 1995年
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Biochemical Journal
A cDNA encoding a novel rat mast-cell proteinase (MCP) named rMCP-3 was successfully cloned and sequenced from the peritoneal cells of Lewis rats infected with the intestinal nematode Nippostrongylus brasiliensis by using the combination of reverse transcription-PCR and rapid-amplification-of-cDNA-ends ('RACE') methods. The cDNA was 979 bp long and included a 741 bp open reading frame. When the deduced amino acid sequence was compared with those of other known mast-cell proteinases, rMCP-3 was considered to be translated as a preproenzyme with a 19-amino-acid signal peptide, a two-amino-acid activation peptide and a 226-amino-acid mature enzyme. The amino acid identity in the mature enzyme was 52.9% and 55.1% with rMCP-1 and rMCP-2 respectively. The rMCP-3 mRNA was not detected in the peritoneal cells of mast-cell-deficient Ws/Ws rats, though it was strongly detected in those of littermate +/+ and Lewis rats, indicating the mast-cell origin of rMCP-3. In addition to being present in peritoneal mast cells, the rMCP-3 mRNA was strongly detected in the skin, tongue, and RBL2H3 rat basophilic leukaemia cells and weakly in the jejunum of N. brasiliensis-infected rats by RNA blot analysis using a rMCP-3 gene-specific probe. By reverse transcription-PCR, the rMCP-3 mRNA was also detected in the lung. While the expression of rMCP-1 and rMCP-2 are clearly restricted in connective-tissue mast cells and mucosal mast cells respectively, rMCP-3 was widely expressed in both types of mast cells with a predominance in connective-tissue mast cells.
DOI： 10.1042/bj3110675
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Seguchi K., Matsuno M., Kataoka H., Kobayashi T., Maruyama H., Itoh H., Koono M., Nawa Y.
American Journal of Tropical Medicine and Hygiene 53 ( 3 ) 263 - 266 1995年
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：American Journal of Tropical Medicine and Hygiene
Gnathostomiasis is primarily a disease of the skin characterized as creeping eruption or mobile erythema. However, larval Gnathostoma sometimes migrate into an unexpected site to elicit serious illness. Here we describe a case of colonic ileus caused by Gnathostoma doloresi. The patient was a 57- year-old man living in Miyazaki Prefecture, Japan, which is known as an area endemic for this parasite. One week after having eaten a few slices of the flesh of a snake (Agkistrodon halys), he developed severe abdominal pain. An abdominal radiograph revealed multiple gas-fluid levels with a distended bowel of an inverted U shape. A barium enema revealed a tumor in the ascending colon near the hepatic flexure that was surgically removed by simple colonic resection. An oblique section of a parasite surrounded by massive infiltration of eosinophils was found by postoperative histopathologic examination. The entire body of the advanced third-stage larva of G. doloresi was dissected from a specimen-embedded paraffin block.
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Molecular cloning of mouse intestinal trefoil factor and its expression during goblet cell changes
Tomita M., Itoh H., Ishikawa N., Higa A., Ide H., Murakumo Y., Maruyama H., Koga Y., Nawa Y.
Biochemical Journal 311 ( 1 ) 293 - 297 1995年
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Biochemical Journal
A cDNA encoding mouse intestinal trefoil factor (mITF) was successfully cloned and sequenced from the small intestine of C57BL/6 mouse by using the combination of reverse transcription-PCR and rapid amplification of cDNA ends methods. The gene was, similar to rat and human ITFs, mainly expressed in the small and large intestine. The mITF expression was up-regulated during the recovery phase after depletion of goblet cells in acetic acid-induced colitis. On the other hand, the expression in the jejunum was not altered, while goblet cell hyperplasia was induced by Nippostrongylus brasiliensis infection. These results suggest that the mITF expression did not simply correlate with the number of goblet cells. The mITF may play an important role in the maintenance and repair of mucosal function of the rectum. Additionally, the mITF in the jejunum may play a role in alteration of the physicochemical nature of goblet cell mucins, thereby affecting the establishment of intestinal helminths.
DOI： 10.1042/bj3110293
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Herlyn D., Harris D., Zaloudik J., Sperlagh M., Maruyama H., Jacob L., Kieny M., Scheck S., Somasundaram R., Hart E., Ertl H., Mastrangelo M.
Journal of Immunotherapy 15 ( 4 ) 303 - 311 1994年5月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Immunotherapy
Monoclonal anti-idiotypic antibody (Ab2) VF2 was derived from rats immunized with anti-colorectal carcinoma (anti-CRC) monoclonal antibody (Abl) CO17–1A. In rabbits the Ab2 induced anti anti-idiotypic antibodies (Ab3) that shared idiotopes with the Abl, bound to the same epitope on CRC cells as Abl, and bound to the isolated CO17–1A antigen. Monoclonal Ab2 VF2 was superior to the previously described polyclonal goat Ab2 against Abl CO17–1A in its capacity to elicit humoral immunity in animals. Ab2 VF2 also induced a specific delayed-type hypersensitivity (DTH) response to challenge with irradiated CO17–1A antigen-positive human CRC cells in mice. Of nine CRC patients immunized with aluminum hydroxide-precipitated Ab2 VF2, six developed antibodies that bound to Ab2, but only three patients developed Ab3 that bound to idiotypic determinants on Ab2. However, the Ab3 did not bind to CO17–1A antigen-positive CRC cells. In contrast, in a previously described trial with polyclonal goat Ab2 to Abl CO17–1A, most of the patients developed anti-CRC antibodies. Four of the nine patients immunized with Ab2 VF2 developed DTH responses to intradermal challenge with the Ab2, and in one patient DTH was both Ab2− and antigen-specific. Peripheral blood mononu-clear cells of the four DTH-reactive patients did not proliferate in response to in vitro stimulation with either Ab2 or antigen. These studies demonstrate that the immunomodulatory activity of monoclonal Ab2 VF2 in animals is only in part predictive of its activity in patients. © 1984 Journal of Immunotherapy. All rights reserved.
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Polyclonal antiidiotypic antibodies mimicking gp 120 of HIV-1
Sperlagh M., Hoxie J., Maruyama H., Stefano K., Gonzales-Scarano F., Prewett M., Liang S., Matsushita S., Herlyn D.
Viral Immunology 7 ( 2 ) 61 - 69 1994年
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Viral Immunology
Rabbit antiidiotypic antibodies (Ab2) were produced against anti-HIV-1 antibody 0.5β (Ab1), which binds to gp120 of HIV-1 and shows virus-neutralizing activity. The Ab2 bound specifically to the Ab1 and their binding to Ab2 was inhibited by a recombinant fragment of gp120 (PB1) or a peptide (residues 301-324 of gp120), both expressing the Ab1-defined epitope. The Ab2 induced in rats antiantiidiotypic antibodies (Ab3) that were Ab1-like in their binding reactivities to PB1, native gp120 or peptide and shared idiotopes with the Ab1. However, the Ab2 did not induce virus-neutralizing Ab3, probably a reflection of the low avidity of the Ab3 as compared to the Ab1.
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Sperlagh M., Stefano K., Gonzalez-Scarano F., Liang S., Hoxie J., Maruyama H., Prewett M., Matsushita S., Herlyn D.
AIDS 7 ( 12 ) 1553 - 1559 1993年12月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：AIDS
Objective: To develop effective, specific and safe anti-idiotypic antibody (Ab2) vaccines against HIV-1. Design: Murine monoclonal Ab2 were generated against anti-HIV-1 antibody 0.5β (Ab1), which binds to gp120, neutralizes HIV-1 and inhibits virus-induced syncytia formation. Methods: Mice were immunized with Ab1, and Ab2 were produced from immunized mice by the hybridoma technique. The Ab2 were characterized in vitro, injected into rabbits, and the anti-anti-idiotypes (Ab3) induced in the rabbits were analyzed for binding and antiviral reactivities by enzyme-linked immunosorbent assay, p24gag release and syncytia formation assays. Results: Seven Ab2 bound to the antigen-combining site of Ab1, one of which (UD7) induced Ab3 in rabbits that were Ab1-like in their binding reactivities to PB1 (recombinant gp120 fragment) or peptides of gp120, and shared idiotypes with the Ab1. Crude Ab3-containing sera specifically and effectively neutralized the virus. Conclusion: Monoclonal Ab2 UD7 has potential as a vaccine against HIV-1.
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Makoto O., Shiroh F., Kouki K., Yoichiro H., Haruhiko M., Hiromi H., Yukifumi N.
Molecular Immunology 30 ( 14 ) 1315 - 1320 1993年10月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Molecular Immunology
To compare the molecular structure of a parasite-derived neutrophil chemotactic factor (NCF) with host-derived NCFs or other NCFs, molecular cloning of cDNA encoding NCF derived from Dirofilaria immitis adult worm (DiNCF) was performed. A D. immitis cDNA library was screened with an antibody to DiNCF, and one DiNCF cDNA clone (pD-4) was isolated. A fusion protein of pD-4 and gene 10 protein showed significant neutrophil chemotactic activity whereas gene 10 protein itself showed marginal neutrophil chemotactic activity. The total nucleotide sequence analysis revealed that pD-4 was 994 bp long with a 432 bp open leading frame encoding a 143 residue protein. The NH 2 -terminal amino acid sequence of the natural DiNCF and the deduced amino acid sequence from the cDNA showed that the mature functional protein was comprised of 112 amino acids. Although the deduced amino acid sequence of this protein did not show overall homology to host-derived NCFs or other known proteins, it contained a similar sequence (Met-Phe-Lys) to the known chemotactic peptides. The possibility of the functional epitope of DiNCF is discussed. © 1993.
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Levi M., Sällberg M., Rudén U., Herlyn D., Maruyama H., Wigzell H., Marks J., Wahren B.
Proceedings of the National Academy of Sciences of the United States of America 90 ( 10 ) 4374 - 4378 1993年5月
記述言語：英語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Proceedings of the National Academy of Sciences of the United States of America
A complementarity-determining region (CDR) of the mouse monoclonal antibody (mAb) F58 was constructed with specificity to a neutralization-inducing region of human immunodeficiency virus type 1 (HIV-1). The mAb has its major reactivity to the amino acid sequence I - GPGRA in the V3 viral envelope region. All CDRs including several framework amino acids were synthesized from the sequence deduced by cloning and sequencing mAb F58 heavy- and light-chain variable domains. Peptides derived from the third heavy-chain domain (CDR-H3) alone or in combination with the other CDR sequences competed with F58 mAb for the V3 region. The CDR-H3 peptide was chemically modified by cyclization and then inhibited HIV-1 replication as well as syncytium formation by infected cells. Both the homologous IIIB viral strain to which the F58 mAb was induced and the heterologous SF2 strain were inhibited. This synthetic peptide had unexpectedly potent antiviral activity and may be a potential tool for treatment of HIV-infected persons.
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Molecular cloning of murine monoclonal anti-idiotypic Fab
Kasai Y., Herlyn D., Sperlagh M., Maruyama H., Matsushita S., Linnenbach A.
Journal of Immunological Methods 155 ( 1 ) 77 - 89 1992年10月
記述言語：日本語 掲載種別：研究論文（学術雑誌） 出版者・発行元：Journal of Immunological Methods
Anti-idiotypic antibodies (Ab2) binding to idiotopes on antibodies with various antigen binding specificities (Ab1) are potential regulators of immunity in a variety of diseases, such as autoimmunity, cancer, and viral, bacterial, or parasitic infections. Furthermore, Ab2 are useful probes for the characterization of receptor/ligand interactions. Thus far, Ab2 production has been limited to the isolation of polyclonal Ab2 from immune sera or monoclonal Ab2 from hybridoma supernatants. However, both approaches have produced a limited number of Ab2. As an alternative approach, we demonstrate here the production of Ab2-Fab by using repertoire cloning. Using HIV-1 as a model system, the Ab2-Fab were generated from the spleens of mice immunized with the virus-neutralizing and syncytia-inhibiting anti-HIV-1 monoclonal antibody 0.5β. A bacteriophage γ vector system was used to express a combinatorial library in Escherichia coli. Iodinated 0.5β was used to identify 17 Ab2-Fab clones. DNA sequence analysis of five clones revealed three similar κ and Fd combinations. The Ab2-Fab bound with high affinity (3.5-6.5 × 109 liters/mol) specifically to the Ab1 and not to isotype-matched antibodies with unrelated specificities. The three Ab2-Fab probably bind to the same idiotope on the Ab1 as demonstrated in cross-competition binding studies. The Ab2-Fab inhibited binding of the Ab1 to antigen, and therefore, may functionally mimic the epitope defined by the Ab1. Repertoire cloning of Ab2-Fab may facilitate the generation of Ab2 that have potential as modulators of immune responses against various antigens. © 1992.