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Vol. 48, No. 2-3, 2005
Issue release date: March–June 2005
Intervirology 2005;48:84–88
(DOI:10.1159/000081733)

Removal of Hepatitis C Virus by G-1 Beads in Sera from Patients with Chronic Hepatitis C

Moriyama M.a · Kaneko M.a · Matsumura H.a · Fukushima A.a · Aoki H.a · Tanaka N.a · Kuroda K.b · Shimizu K.b · Hiraishi K.c · Arakawa Y.a
aThird Department of Internal Medicine, and bDepartment of Microbiology and Immunology, Nihon University School of Medicine, Itabashiku, Tokyo, and cJapan Immunoresearch Laboratories, Takasaki, Gunma, Japan
email Corresponding Author

Abstract

Recently, a new method of extracorporeal granulocyte depletion apheresis has been developed to treat inflammatory systemic diseases using an Adacolumn (Japan Immunoresearch Laboratories, Takasaki, Japan) that is filled with acetate cellulose beads (G-1 beads) to adsorb the granulocytes. We examined whether hepatitis C virus (HCV) is adsorbed after incubation of the Adacolumn with the sera from patients with HCV-RNA-positive chronic hepatitis C. Patients and Methods: A total of 10 patients with chronic hepatitis C, whose levels of HCV RNA were greater than 800 kIU/ml were examined. The serum was incubated with 500 G-1 beads in a syringe at 37°C for 1 h. After removal of the serum, the beads were washed with RNase-free water. The G-1 beads were removed from the syringe after centrifugation. RNA was extracted from 200 µl of the wash waste and from 10, 50, 100 and 200 beads, respectively, using TRIZol regent. Detection of HCV RNA was performed using the nested PCR method. Results: HCV RNA was detected from as few as 10 G-1 beads. HCV RNA was not detected from waste fluid collected after the last wash from any of the patients. Further, HCV RNA was detected in the initial waste fluid after the 37°C incubation with serum in all of the patients. Since HCV RNA was detected on the G-1 beads, but not from the last washing solution in the current examination, these results suggest that the G-1 beads adsorbed HCV RNA. Conclusions: Our in vitro study confirmed that G-1 beads adsorbed HCV; therefore, apheresis using a column filled with G-1 beads may reduce the HCV RNA load in the blood of patients with chronic hepatitis C.


 goto top of outline Key Words

  • G-1 beads
  • Adacolumn
  • Hepatitis C virus RNA
  • Chronic hepatitis C
  • Eradication of hepatitis C virus
  • Extracorporeal granulocyte apheresis

 goto top of outline Abstract

Recently, a new method of extracorporeal granulocyte depletion apheresis has been developed to treat inflammatory systemic diseases using an Adacolumn (Japan Immunoresearch Laboratories, Takasaki, Japan) that is filled with acetate cellulose beads (G-1 beads) to adsorb the granulocytes. We examined whether hepatitis C virus (HCV) is adsorbed after incubation of the Adacolumn with the sera from patients with HCV-RNA-positive chronic hepatitis C. Patients and Methods: A total of 10 patients with chronic hepatitis C, whose levels of HCV RNA were greater than 800 kIU/ml were examined. The serum was incubated with 500 G-1 beads in a syringe at 37°C for 1 h. After removal of the serum, the beads were washed with RNase-free water. The G-1 beads were removed from the syringe after centrifugation. RNA was extracted from 200 μl of the wash waste and from 10, 50, 100 and 200 beads, respectively, using TRIZol regent. Detection of HCV RNA was performed using the nested PCR method. Results: HCV RNA was detected from as few as 10 G-1 beads. HCV RNA was not detected from waste fluid collected after the last wash from any of the patients. Further, HCV RNA was detected in the initial waste fluid after the 37°C incubation with serum in all of the patients. Since HCV RNA was detected on the G-1 beads, but not from the last washing solution in the current examination, these results suggest that the G-1 beads adsorbed HCV RNA. Conclusions: Our in vitro study confirmed that G-1 beads adsorbed HCV; therefore, apheresis using a column filled with G-1 beads may reduce the HCV RNA load in the blood of patients with chronic hepatitis C.

Copyright © 2005 S. Karger AG, Basel


 goto top of outline References
  1. Kyogoku M, Kasukawa R: Clinical and basic studies on the G-1 column, a new extracorporeal therapeutic device effective in controlling rheumatoid arthritis. Inflamm Res 1998;47(suppl 3):S166–S176.

    External Resources

  2. Ishida H, Ohara M, Watanabe S, Takase Y, Kasukawa R: Treatment of rheumatoid arthritis by granulo-monocyteaphresis. Jpn J Rheumatol 1996;6:113–124.
  3. Nagashima M, Yoshino S, Tanaka H, Yoshida N, Kashiwagi N, Saniabadi AR: Granulocyte and monocyte apheresis suppresses symptoms of rheumatoid arthritis: A pilot study. Rheumatol Int 1998;18:113–118.
  4. Tanaka S, Kashiwagi N, Adachi M, Asao T, Nagamachi Y: Efficacy of extracorporeal granulotrap column (G-1 column) in rabbit endotoxin shock models. Kitakanto Med J 1997;47:393–398.

    External Resources

  5. Fujimori J, Yoshino S, Koiwa M, Hirai H, Shiga H, Hayama N, Iino Y: Improvement in rheumatoid arthritis following application of an extracorporeal granulotrap column, G-1. Rheumatol Int 1996;15:175–180.
  6. Kashiwagi N, Sugimura K, Koiwai H, Yamamoto H, Yoshikawa T, Saniabadi AR, Adachi M, Shimoyama T: Immunomodulatory effects of granulocyte and monocyte adsorption apheresis as a treatment for patients with ulcerative colitis. Dig Dis Sci 2002;47:1334–1341.
  7. Shimoyama T, Sawada K, Hiwatashi N, Sawada T, Matsueda K, Munakata A, Asakura H, Tanaka T, Kasukawa R, Kimura K, Suzuki Y, Nagamachi Y, Muto T, Nagawa H, Iizuka B, Baba S, Nasu M, Kataoka T, Kashiwagi N, Saniabadi AR: Safety and efficacy of granulocyte and monocyte adsorption apheresis in patients with active ulcerative colitis: A multicenter study. J Clin Apheresis 2001;16:1–9.
  8. Beretta A, Clerici M, Hasson H, Fumagalli L, Trabattoni D, Lillo F, Ferrante P, Saniabadi AR, Adachi M, Lazzarin A: Ex-vivo purging of circulating monocytes results in immunovirologic improvement in partially HAART responder HIV-infected patients. J Biol Regul Homeost Agents 2000;14:27–31.
  9. Nirei K, Kaneko M, Moriyama M, Arakawa Y: The clinical feature of chronic hepatitis C are not affected by the coexistence of hepatitis B virus DNA in patients negative for hepatitis B virus surface antigen. Intervirology 2000;43:95–101.
  10. Shimizu T, Moriyama M, Arakawa Y: Detection of HGV RNA in digestive organs. Intervirology 2001;44:14–20.
  11. Sioda A, Moriyama M, Matsumura H, Kaneko M, Tanaka N, Arakawa Y: Clinicopathological features of serum TTV DNA-positive non-A to G liver diseases in Japan. Hepatol Res 2001;21:169–180.
  12. Mikuni M, Moriyama M, Tanaka N, Abe K, Arakawa Y: SEN virus infection does not affect the progression of non-A to -E liver disease. J Med Virol 2002;67:624–629.
  13. Inchauspe G, Abe K, Zebedee S, Nasoff M, Prince AM: Use of conserved sequences from hepatitis C virus for the detection of viral RNA in infected sera by polymerase chain reaction. Hepatology 1991;14:595–600.
  14. Matsumura H, Moriyama M, Tanaka N, Ohkubo H, Arakawa Y: The serum hepatitis C virus RNA at two weeks after the administration of interferon: Its predictability for the efficacy of interferon treatment for chronic hepatitis C. Nihon Univ J Med 1998;40:61–69.
  15. Okamoto H, Sugiyama Y, Okada S, Kurai K, Akahane Y, Sugai Y, Tanaka T, Tanaka T, Sato K, Tsuda F, Miyakawa Y, Mayumi M: Typing of hepatitis C virus by PCR with type-specific primers: Application to clinical surveys and tracing infectious sources. J Gen Virol 1992;73:673–679.
  16. Simmonds P, Alberti A, Alter HJ, Bonino F, Bradley DW, Brechot C, Brouwer JT, Chan SW, Chayama K, Chen DS, Choo QL, Colombo M, Cuypers HTM, Date T, Dusheiko GM, Esteban JI, Fay O, Hadziyannis SJ, Han J, Hatzakis A, Holmes EC, Hotta H, Houghton M, Irvine B, Kohara M, Kolberg JA, Kuo G, Lau JYN, Lelie PN, Maertens G, McOmish F, Miyamura T, Mizokami M, Nomoto A, Prince AM, Reesink HW, Rice C, Roggendorf M, Schalm SW, Shikata T, Shimotohno K, Stuyver L, Trepo C, Weiner A, Yap PL, Urdea MS: A proposed system for the nomenclature of hepatitis C viral genotypes. Hepatology 1994;19:1321–1324.
  17. Enomoto N, Sakuma I, Asahina Y, Kurosaki M, Murakami T, Yamamoto C, Ogura Y, Izumi N, Marumo F, Sato C: Mutations in the nonstructural protein 5A gene and response to interferon in patients with chronic hepatitis C virus 1b infection. N Engl J Med 1996;334:77–81.

 goto top of outline Author Contacts

Mitsuhiko Moriyama, MD, PhD
Third Department of Internal Medicine, Nihon University School of Medicine
30-1 Oyaguchi Kamimachi, Itabashiku, Tokyo 173-8610 (Japan)
Tel. +81 3 3972 8111, ext. 2426, Fax +81 3 3956 8496
E-Mail moriyama@med.nihon-u.ac.jp


 goto top of outline Article Information

Received: October 10, 2003
Accepted after revision: March 2, 2004
Number of Print Pages : 5
Number of Figures : 3, Number of Tables : 0, Number of References : 17


 goto top of outline Publication Details

Intervirology (International Journal of Basic and Medical Virology)

Vol. 48, No. 2-3, Year 2005 (Cover Date: March-June 2005)

Journal Editor: U.G. Liebert, Leipzig
ISSN: 0300–5526 (print), 1423–0100 (Online)

For additional information: http://www.karger.com/int


Copyright / Drug Dosage / Disclaimer

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

Abstract

Recently, a new method of extracorporeal granulocyte depletion apheresis has been developed to treat inflammatory systemic diseases using an Adacolumn (Japan Immunoresearch Laboratories, Takasaki, Japan) that is filled with acetate cellulose beads (G-1 beads) to adsorb the granulocytes. We examined whether hepatitis C virus (HCV) is adsorbed after incubation of the Adacolumn with the sera from patients with HCV-RNA-positive chronic hepatitis C. Patients and Methods: A total of 10 patients with chronic hepatitis C, whose levels of HCV RNA were greater than 800 kIU/ml were examined. The serum was incubated with 500 G-1 beads in a syringe at 37°C for 1 h. After removal of the serum, the beads were washed with RNase-free water. The G-1 beads were removed from the syringe after centrifugation. RNA was extracted from 200 µl of the wash waste and from 10, 50, 100 and 200 beads, respectively, using TRIZol regent. Detection of HCV RNA was performed using the nested PCR method. Results: HCV RNA was detected from as few as 10 G-1 beads. HCV RNA was not detected from waste fluid collected after the last wash from any of the patients. Further, HCV RNA was detected in the initial waste fluid after the 37°C incubation with serum in all of the patients. Since HCV RNA was detected on the G-1 beads, but not from the last washing solution in the current examination, these results suggest that the G-1 beads adsorbed HCV RNA. Conclusions: Our in vitro study confirmed that G-1 beads adsorbed HCV; therefore, apheresis using a column filled with G-1 beads may reduce the HCV RNA load in the blood of patients with chronic hepatitis C.



 goto top of outline Author Contacts

Mitsuhiko Moriyama, MD, PhD
Third Department of Internal Medicine, Nihon University School of Medicine
30-1 Oyaguchi Kamimachi, Itabashiku, Tokyo 173-8610 (Japan)
Tel. +81 3 3972 8111, ext. 2426, Fax +81 3 3956 8496
E-Mail moriyama@med.nihon-u.ac.jp


 goto top of outline Article Information

Received: October 10, 2003
Accepted after revision: March 2, 2004
Number of Print Pages : 5
Number of Figures : 3, Number of Tables : 0, Number of References : 17


 goto top of outline Publication Details

Intervirology (International Journal of Basic and Medical Virology)

Vol. 48, No. 2-3, Year 2005 (Cover Date: March-June 2005)

Journal Editor: U.G. Liebert, Leipzig
ISSN: 0300–5526 (print), 1423–0100 (Online)

For additional information: http://www.karger.com/int


Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

References

  1. Kyogoku M, Kasukawa R: Clinical and basic studies on the G-1 column, a new extracorporeal therapeutic device effective in controlling rheumatoid arthritis. Inflamm Res 1998;47(suppl 3):S166–S176.

    External Resources

  2. Ishida H, Ohara M, Watanabe S, Takase Y, Kasukawa R: Treatment of rheumatoid arthritis by granulo-monocyteaphresis. Jpn J Rheumatol 1996;6:113–124.
  3. Nagashima M, Yoshino S, Tanaka H, Yoshida N, Kashiwagi N, Saniabadi AR: Granulocyte and monocyte apheresis suppresses symptoms of rheumatoid arthritis: A pilot study. Rheumatol Int 1998;18:113–118.
  4. Tanaka S, Kashiwagi N, Adachi M, Asao T, Nagamachi Y: Efficacy of extracorporeal granulotrap column (G-1 column) in rabbit endotoxin shock models. Kitakanto Med J 1997;47:393–398.

    External Resources

  5. Fujimori J, Yoshino S, Koiwa M, Hirai H, Shiga H, Hayama N, Iino Y: Improvement in rheumatoid arthritis following application of an extracorporeal granulotrap column, G-1. Rheumatol Int 1996;15:175–180.
  6. Kashiwagi N, Sugimura K, Koiwai H, Yamamoto H, Yoshikawa T, Saniabadi AR, Adachi M, Shimoyama T: Immunomodulatory effects of granulocyte and monocyte adsorption apheresis as a treatment for patients with ulcerative colitis. Dig Dis Sci 2002;47:1334–1341.
  7. Shimoyama T, Sawada K, Hiwatashi N, Sawada T, Matsueda K, Munakata A, Asakura H, Tanaka T, Kasukawa R, Kimura K, Suzuki Y, Nagamachi Y, Muto T, Nagawa H, Iizuka B, Baba S, Nasu M, Kataoka T, Kashiwagi N, Saniabadi AR: Safety and efficacy of granulocyte and monocyte adsorption apheresis in patients with active ulcerative colitis: A multicenter study. J Clin Apheresis 2001;16:1–9.
  8. Beretta A, Clerici M, Hasson H, Fumagalli L, Trabattoni D, Lillo F, Ferrante P, Saniabadi AR, Adachi M, Lazzarin A: Ex-vivo purging of circulating monocytes results in immunovirologic improvement in partially HAART responder HIV-infected patients. J Biol Regul Homeost Agents 2000;14:27–31.
  9. Nirei K, Kaneko M, Moriyama M, Arakawa Y: The clinical feature of chronic hepatitis C are not affected by the coexistence of hepatitis B virus DNA in patients negative for hepatitis B virus surface antigen. Intervirology 2000;43:95–101.
  10. Shimizu T, Moriyama M, Arakawa Y: Detection of HGV RNA in digestive organs. Intervirology 2001;44:14–20.
  11. Sioda A, Moriyama M, Matsumura H, Kaneko M, Tanaka N, Arakawa Y: Clinicopathological features of serum TTV DNA-positive non-A to G liver diseases in Japan. Hepatol Res 2001;21:169–180.
  12. Mikuni M, Moriyama M, Tanaka N, Abe K, Arakawa Y: SEN virus infection does not affect the progression of non-A to -E liver disease. J Med Virol 2002;67:624–629.
  13. Inchauspe G, Abe K, Zebedee S, Nasoff M, Prince AM: Use of conserved sequences from hepatitis C virus for the detection of viral RNA in infected sera by polymerase chain reaction. Hepatology 1991;14:595–600.
  14. Matsumura H, Moriyama M, Tanaka N, Ohkubo H, Arakawa Y: The serum hepatitis C virus RNA at two weeks after the administration of interferon: Its predictability for the efficacy of interferon treatment for chronic hepatitis C. Nihon Univ J Med 1998;40:61–69.
  15. Okamoto H, Sugiyama Y, Okada S, Kurai K, Akahane Y, Sugai Y, Tanaka T, Tanaka T, Sato K, Tsuda F, Miyakawa Y, Mayumi M: Typing of hepatitis C virus by PCR with type-specific primers: Application to clinical surveys and tracing infectious sources. J Gen Virol 1992;73:673–679.
  16. Simmonds P, Alberti A, Alter HJ, Bonino F, Bradley DW, Brechot C, Brouwer JT, Chan SW, Chayama K, Chen DS, Choo QL, Colombo M, Cuypers HTM, Date T, Dusheiko GM, Esteban JI, Fay O, Hadziyannis SJ, Han J, Hatzakis A, Holmes EC, Hotta H, Houghton M, Irvine B, Kohara M, Kolberg JA, Kuo G, Lau JYN, Lelie PN, Maertens G, McOmish F, Miyamura T, Mizokami M, Nomoto A, Prince AM, Reesink HW, Rice C, Roggendorf M, Schalm SW, Shikata T, Shimotohno K, Stuyver L, Trepo C, Weiner A, Yap PL, Urdea MS: A proposed system for the nomenclature of hepatitis C viral genotypes. Hepatology 1994;19:1321–1324.
  17. Enomoto N, Sakuma I, Asahina Y, Kurosaki M, Murakami T, Yamamoto C, Ogura Y, Izumi N, Marumo F, Sato C: Mutations in the nonstructural protein 5A gene and response to interferon in patients with chronic hepatitis C virus 1b infection. N Engl J Med 1996;334:77–81.