Jpn. J. Infect. Dis., 52, 1999
Laboratory and Edpidemiology Communications
Prevalence of Drug Resistance-Related Mutations among HIV-1s in Japan
Wataru Sugiura*, Masakazu Matsuda, Hanae Abumi, Kaneo Yamada 1, Masashi Taki 2, Masaaki Ishikawa 3,Takuma Miura 4, Katsuyuki Fukutake 5, Kengo Gouchi 6, Atsushi Ajisawa 7, Aikichi Iwamoto 8,Hideji Hanabusa 9, Junichi Mimaya 10, Junki Takamatsu 11, Noboru Takada 12, Eizo Kakishita 13,Akira Yoshioka 14, Seizaburou Kashiwagi 15, Akira Shirahata 16 and Yoshiyuki Nagai
AIDS Research Center, National Institute of Infectious Diseases, 4-7-1, Gakuen, Musashimurayama-shi, Tokyo,1Institute of Medical Science, St. Marianna University School of Medicine, 2-6-1, Sugao, Miyamae-ku, Kawasaki-shi, Kanagawa, 2Dept. of Pediatrics, St. Marianna University, School of Medicine, 2-6-1, Sugao, Miyamae-ku, Kawasaki-shi, Kanagawa, 3Third Department of Internal Medicine, Tohoku University, School of Medicine, 1-1, Seiryo-cho, Aoba-ku, Sendai-shi, Miyagi, 4Department of Pediatrics, Haga Red Cross Hospital, 2461, Dai-machi, Mooka-shi, Tochigi, 5Department of Clinical Pathology, Tokyo Medical University, 6-7-1, Nishishinjuku, Shinjuku-ku, Tokyo, 6Department of Internal Medicine, Teikyo University, School of Medicine, 2-11-1, Kaga, Itabashi-ku, Tokyo, 7Department of Infectious Diseases, Tokyo Metropolitan Komagome Hospital, 3-18-22, Honkomagome, Bunkyo-ku, Tokyo, 8Department of Infectious Diseases, Institute of Medical Science, The University of Tokyo, 4-6-1, Shiroganedai, Minato-ku, Tokyo, 9Department of Hematology, Ogikubo Hospital, 3-1-24, Imagawa, Suginami-ku, Tokyo, 10Division of Hematology- Oncology, Children's Hospital of Shizuoka Prefecture, 860, Urushiyama, Shizuoka-shi, Shizuoka, 11Department of Blood Transfusion, Nagoya University Hospital, 65, Tsurumai, Syowa-ku, Nagoya-shi, Aichi, 12Division of Blood Transfusion Services, HiroshimaUniversity Medical Hospital, 1-2-3, Kasumi, Minami-ku, Hiroshima-shi, Hiroshima, 13 Second Department of Internal Medicine, Hyogo College of Medicine, 1-1, Mukogawa-cho, Nishinomiya-shi, Hyogo, 14Department of Pediatrics, Nara Medical College, 840, Shijo-cho, Kashihara-shi, Nara, 15 Department of General of Medicine, Kyushu University Hospital, 3-1-1, Maidashi, Higashi-ku, Fukuoka, 16 Department of Pediatrics, University of Occupational and Environmental Health, 1-1, Iseigaoka, Yahatanishi-ku, Kitakyushu-shi, Fukuoka.
Communicated by Hiroshi Yoshikura
(Accepted February 23, 1999)
Nucleoside analogue reverse transcriptase inhibitors (NRTI) and protease inhibitors (PI) began being used in Japan for treatment of HIV-1 infected patients shortly after they were found to be effective (1). Highly Active Anti-Retroviral Therapy (HAART) (2) is now recommended. One of the problems in using these anti-HIV-1 drugs is the appearance of drug-resistant mutant viruses. From November 1996 to December 1998, we examined 1,250 clinical specimens from 329 patients for drug resistance-related mutations. Seventeen hospitals in various parts of Japan participated in this project. The 329 clinical cases consisted of 210 hemophiliacs infected through contaminated blood products (64%), 55 homosexu- ally infected patients (17%), 52 heterosexually infected patients (16%), 2 patients infected through blood transfusion (1%), and 10 patients whose infection route was unknown. 166 cases received three drugs (50%), 106 cases one or two drugs (32%), and 21 cases no drugs; the clinical histories of 36 cases were not known. Among the anti-HIV-1 drug-treated patients, 245 patients received AZT (Zidovudine), 134 patients ddI (Didanosine), 101 patients ddC (Zalcitabine), 125 patients d4T (Stavudine), 179 patients 3TC (Lamivudine), 81 patients IDV (Indinavir), 38 patients SQV (Saquinavir), 26 patients RTV (Ritonavir), and 89 patients NFV (Nelfinavir). AZT, ddI, d4T, ddC, and 3TC are NRTIs, and IDV, RTV, SQV, and NFV are PIs. For determining drug resistance-related mutations, we used the Los Alamos HIV-1 Drug Resistant Database (3). The data presented below are based on the last data obtained for each patient. Viral RNAs were extracted from 200ƒÊl plasma by using an RNA purification kit (Boehringer Mannheim, Indianapolis, IN), and the reverse transcriptase (RT) and protease (PR) regions were RT/PCR-amplified in the manner reported previously (4). The amplified RT region was 380bp long and the RT region 821 bp long. Sequencing by the dye-terminator method (5) was conducted by using ABI-377 (Applied Biosystems, Foster City, CA) and Sequence Navigator software (Applied Biosystems). Figs. 1A and B show the incidence of mutants found in the patients treated with anti-HIV-1 drugs. Among viruses derived from AZT-treated patients, AZT-specific mutation T215Y/E was the most frequent (near 40%), followed by AZT-specific mutations M41L, D67N, and K70R. Among viruses derived from 3TC-treated patients, 53% had 3TC-specific mutation M184V. Among viruses derived from PI-treated patients, mutation L90M for SQV resistance was the most frequent (32%), followed by mutations D30N for NFV resistance, G48V for SQV resistance, M46I/L for IDV resistance, and V82A/F/T for IDV and/or RTV resistance. The mutations occurred only in patients treated with the respective drugs and never in those treated with other drugs (Fig. 1A). Drug resistance to AZT or 3TC was found in 50% of the patients treated with the respective drugs. Viruses with mutations related to resistance to ddI or d4T were far less frequent (4-6%) (Fig. 1B). Fig. 2 shows the incidence of the drug resistant viruses in all specimens including those from untreated patients. The resistance to AZT and that to 3TC occurred in 38% and 28% of the specimens, respectively. Among the viruses derived from patients who had not been treated with anti-HIV-drugs, the primary mutations were few. There were no NRTI resistance-related mutations and a very few PI resistance-related mutations (V82A/F/T for IDV and G48V for SQV in 1%). PI resistance-related secondary mutations were more frequent; mutation L10I/R/V was found in 2%, K20M/R in 4%, M36I in 9.2%, L63P in 42%, and A71V/T in 9.2%. Fig. 3 shows the incidence of viruses classified according to number of drugs to which the viruses were considered resistant. About half of the viruses had mutation(s) for resistance against at least one drug, and one-third had mutations for resistance against two or more drugs. One case showed resistance even to seven drugs (Fig. 3A). Half of the viruses had mutations for resistance against at least one NRTI, and a quarter had mutations for resistance against two or more NRTIs (Fig. 3B). The incidence of mutations related to PI resistance was lower; 9% of the viruses had mutations for resistance against two or more PIs (Fig. 3C). The data as a whole suggested a high prevalence of anti-HIV drug resistant mutants among HIV-1s in Japanese patients. This study was supported by the Organization for Pharmaceutical Safety and Research (OPSR) of Japan. REFERENCES 1. Hirsh, M.S. (1997): Current antiretrovirals-a review. Antiviral Therapy, 2(suppl. 4), 19-40. 2. Palella, F.J.Jr., Delaney, K.M., Moorman, A.C., Loveless, M.O., Fuhrer, J., Satten, G.A., Aschman, D.J., Holmberg, S.D. and HIV Outpatient Study Investigators (1998): Declining morbidity and mortality among patients with advanced human immuno-deficiency virus infection. N. Engl. J. Med., 338, 853-860. 3. Jennifer, H., Brendan, A.L., Raymond, F.S. and John, W.M.(1997): Mutations in Retroviral Genes Associated with Drug Resistance. Human Retroviruses and AIDS, III-207-249. 4. Kitsutani,P., Naganawa, S., Shiino,T., Matsuda, M., Honda, M., Yamada, K., Taki, M. and Sugiura,W. (1998): HIV-1 Type 1 Subtypes of Nonhemophiliac Patients in Japan. AIDS Res. Hum. Retroviruses, 14, 1099-1103. 5. Sanger, F., Nicklen, S., and Coulson, A.R. (1977): DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. U.S.A., 74, 5463-5467.
* Corresponding author: E-mail:wsugiura@nih.go.jp, Fax:+81-42-561-7746 Go to JJID Homepage Go to JJID 52(1)