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NEW ISOLATES OF HFRS VIRUS IN SICHUAN, CHINA AND CHARACTERISATION OF ANTIGENIC DIFFERENCES BY MONOCLONAL ANTIBODIES
1328 her first months of life, and pipamperone was prescribed (40 mg daily). Her adoptive parents then observed a significant improvement in her skin, with less severe and smaller bullae. During 1985, many trials showed that the disease was dosedependent, with good control of the disease at a dose of 70 mg daily and partial control at 40 mg. Two attempts at drug withdrawal were followed by major bullous eruptions, which led the parents to reintroduce pipamperone. We noted the appearance of a palmoplantar keratoderma under the treatment. Epidermolytic types of epidermolysis bullosa are treated and no drug controls these diseases symptomatically satisfactorily.22 In this child pipamperone controlled epidermolysis bullosa herpetiformis and may be able to cure the condition, as indicated by the appearance of palmoplantar keratoderma. This fact and the dose response observed suggest a causal relation between the drug and the structural defect of the basal cells. Pipamperone is a member of the butyrophenone family and is a neuroleptic agent; since large doses are necessary to control the bullous disorder patients will need to be assessed carefully because of the risk of the malignant neuroleptic
SPECIFIC REACTION PATTERNS ON CHINESE HFRS VIRUS STRAINS IDENTIFIED BY IFA USING MONOCLONAL ANTIBODIES AGAINST HANTAAN VIRUS
syndrome. Departments of Dermatology and Paediatrics, Chu Dupuytren, 87042 Limoges, France 1.
J. M. BONNETBLANC J. J. BOUQUIER
Anton-Lamprecht I, Schnyder HW. Epidermolysis bullosa herpetiformis Dowling Meara: Report of a case and pathomorphogenesis. Dermatologica
1982; 164: 211-35. 2. Haber RM, Hanna W, Ramsay CA, Boxall LBH. Hereditary epidermolysis bullosa. J Am Acad Dermatol 1985; 13: 252-78.
NEW ISOLATES OF HFRS VIRUS IN SICHUAN, CHINA AND CHARACTERISATION OF ANTIGENIC DIFFERENCES BY MONOCLONAL
ANTIBODIES
SIR,-In China notifications of haemorrhagic fever with renal syndrome (HFRS), a disease first described in[ Heilongjiang Province in 1913, have lately been increasing, from 85 293 cases in 1983 to 90 936 in 1984 and 103 778 in 1985.i
In Sichuan Province, where the first case was described in 1960,L than 3000 cases have been reported every year for the past,
more
six years. 4506 lung
specimens from thirteen species of small mammal captured in Sichuan have been studied by the indirect immunofluorescent antibody technique (IFA),1 with a rabbit antibody against strain 76-118 of Korean haemorrhagic fever virus and a sheep anti-rabbit immunoglobulin conjugated to fluorescein isothiocyanate (FITC). HFRS antigen was detected in six rodent species (Apodemus agrarius, Mus musculus, Rattus norvegicus, R flavipectus, R nitidus, and R fulvescins) and two insectivorous species (Anourosorex squamipes and Crocidura russula). A agrarius and R norvegicus the main reservoir hosts. Since 1981, using Vero E6 cells, twenty strains of HFRS virus have been isolated from positive lung specimens of the above mammals (nine from A agrarius, two from R norvegicus, two from R flavipectus, one from A squamipes, one from C russula and five from patients with HFRS in the acute state). The antigenic differences between Chinese HFRS virus strains have not been fully investigated. Some of them were tested only by cross-neutralisation and cross-blocking. Lately, monoclonal antibodies (MoAb) have been used to identify antigenic differences between HFRS virus isolates. 3-6 In our laboratory in Chengdu, a panel of seven MoAbs against Hantaan virus developed by J. B. McC.’s group has been used in the IFA test (Centers for Disease Control standard protocol). The’ Vero E6 cells were infected separately with 76-118 prototype and Chinese strains of HFRS viruses. After passages, the infected cells were applied onto multispot slides, inactivated by ultraviolet rays, and fixed in acetone. The seven MoAbs used were tested at 1:10 working dilution, and Miles FITC anti-mouse IgG at 1:40 diluted in phosphate-buffered saline, pH 72, containing Evans blue at a final concentration of
were
*The
geographical sources of these twenty strains were, in descending order: South
Korea; the provinces of Anhui, Jiangsu, Henan, and Shaanxi (two strains); Linshui (four strains) and Daxian (two strains) counties of Sichuan Province; Chengdu suburb; and Nanchong (five strains) and Linshui (two strains), counties of Sichuan Province.
t..-not tested. 1=FD03-AAll; 2=BDOI-BB08; 3=EC02-BDOl; 4=KDOlAD12 ; 5 = HC02-BD05; 6=HDOl-AD02; 7==JD05-AE04.
0-05%. We used cross-reacting MoAb 25-1 (with a dilution of 1:10) against HFRS A9 strain (Institute of Virology, Chinese Academy of Medical Sciences) as a positive control, and we obtained
positive results on all HFRS virus strains tested. No specific fluorescence was observed when the MoAbs and Miles FITC anti-mouse IgG were added to the Vero E6 cells without infection. In the absence of MoAB, the conjugate did not bind E6 cells infected with HFRS virus strains. The results
to
are
shown in the table. The results suggest that there may be different serotypes of HFRS virus in China as well as in Sichuan Province. This may assist in the selection of reference strains for producing HFRS vaccine. Health and Anti-epidemic Centre of Sichuan Province,
DONG YOU YAN
Chengdu, Sichuan 610031, People’s Republic of China
CHUAN AN ZHANG
YUN JU XIE
Special Pathogens Branch, Division of Viral Diseases, Center for Infectious Diseases, Centers for Disease Control,
Atlanta, Georgia, USA Health and Anti-epidemic Station of Daxiun Prefecture, Sichuan
J. B. MCCORMICK A. SANCHEZ H. M. ENGELMAN SHANG ZHI CHEN
Health and Anti-epidemic Station of Guang-an County, Sichuan
XIAN SHI GU
Health and Anti-epidemic Station of Da County, Sichuan
WEN TING TANG JIAN ZHANG
PW, Johnson KM. Isolation of the etiologic agent of Korean hemorrhagic fever. J Infect Dis 1978; 137: 298-308. Song G, Hang CS, Liao HX, et al. Antigenjc difference between viral strains causing classical and mild types of epidemic hemorrhagic fever with renal syndrome in China. J Inf Dis 1984; 150: 889-94. Franko MC, Gibbs CJ Jr, Lee PW, Gajdusek DC. Monoclonal antibodies specific for Hantaan virus. Proc Natl Acad Sci USA 1983; 80: 4149-53. van der Groen G, Tkachenko EA, Lvanov AP, Lukashevich I, Provost C, McCormick JB. Antigenic differences between Hantaan and USSR strains of HFRS virus using monoclonal antibodies. In: XIth International Congress for Tropical Medicine and Malaria (Calgary, Canada, 1984): abstr 159.
1. Lee HW, Lee 2.
3.
4.
5. Kitamura T. Present status of HFRS studies in the National Institute of Health Tokyo, Japan. Sendai, Japan: WHO Regional Office for the Western Pacific
WRP/CDS/WG/84.11. 1984: 28-31. 6. Chen BQ, Fu JL, Liao HX, et al. The antigenic analysis of EHF viruses by 35 McAbs. Chin J Microbiol Immunol 1984; 5: 136-39 (English abstract).
SPECIFIC REACTION PATTERNS ON CHINESE HFRS VIRUS STRAINS IDENTIFIED BY IFA USING MONOCLONAL ANTIBODIES AGAINST HANTAAN VIRUS
syndrome. Departments of Dermatology and Paediatrics, Chu Dupuytren, 87042 Limoges, France 1.
J. M. BONNETBLANC J. J. BOUQUIER
Anton-Lamprecht I, Schnyder HW. Epidermolysis bullosa herpetiformis Dowling Meara: Report of a case and pathomorphogenesis. Dermatologica
1982; 164: 211-35. 2. Haber RM, Hanna W, Ramsay CA, Boxall LBH. Hereditary epidermolysis bullosa. J Am Acad Dermatol 1985; 13: 252-78.
NEW ISOLATES OF HFRS VIRUS IN SICHUAN, CHINA AND CHARACTERISATION OF ANTIGENIC DIFFERENCES BY MONOCLONAL
ANTIBODIES
SIR,-In China notifications of haemorrhagic fever with renal syndrome (HFRS), a disease first described in[ Heilongjiang Province in 1913, have lately been increasing, from 85 293 cases in 1983 to 90 936 in 1984 and 103 778 in 1985.i
In Sichuan Province, where the first case was described in 1960,L than 3000 cases have been reported every year for the past,
more
six years. 4506 lung
specimens from thirteen species of small mammal captured in Sichuan have been studied by the indirect immunofluorescent antibody technique (IFA),1 with a rabbit antibody against strain 76-118 of Korean haemorrhagic fever virus and a sheep anti-rabbit immunoglobulin conjugated to fluorescein isothiocyanate (FITC). HFRS antigen was detected in six rodent species (Apodemus agrarius, Mus musculus, Rattus norvegicus, R flavipectus, R nitidus, and R fulvescins) and two insectivorous species (Anourosorex squamipes and Crocidura russula). A agrarius and R norvegicus the main reservoir hosts. Since 1981, using Vero E6 cells, twenty strains of HFRS virus have been isolated from positive lung specimens of the above mammals (nine from A agrarius, two from R norvegicus, two from R flavipectus, one from A squamipes, one from C russula and five from patients with HFRS in the acute state). The antigenic differences between Chinese HFRS virus strains have not been fully investigated. Some of them were tested only by cross-neutralisation and cross-blocking. Lately, monoclonal antibodies (MoAb) have been used to identify antigenic differences between HFRS virus isolates. 3-6 In our laboratory in Chengdu, a panel of seven MoAbs against Hantaan virus developed by J. B. McC.’s group has been used in the IFA test (Centers for Disease Control standard protocol). The’ Vero E6 cells were infected separately with 76-118 prototype and Chinese strains of HFRS viruses. After passages, the infected cells were applied onto multispot slides, inactivated by ultraviolet rays, and fixed in acetone. The seven MoAbs used were tested at 1:10 working dilution, and Miles FITC anti-mouse IgG at 1:40 diluted in phosphate-buffered saline, pH 72, containing Evans blue at a final concentration of
were
*The
geographical sources of these twenty strains were, in descending order: South
Korea; the provinces of Anhui, Jiangsu, Henan, and Shaanxi (two strains); Linshui (four strains) and Daxian (two strains) counties of Sichuan Province; Chengdu suburb; and Nanchong (five strains) and Linshui (two strains), counties of Sichuan Province.
t..-not tested. 1=FD03-AAll; 2=BDOI-BB08; 3=EC02-BDOl; 4=KDOlAD12 ; 5 = HC02-BD05; 6=HDOl-AD02; 7==JD05-AE04.
0-05%. We used cross-reacting MoAb 25-1 (with a dilution of 1:10) against HFRS A9 strain (Institute of Virology, Chinese Academy of Medical Sciences) as a positive control, and we obtained
positive results on all HFRS virus strains tested. No specific fluorescence was observed when the MoAbs and Miles FITC anti-mouse IgG were added to the Vero E6 cells without infection. In the absence of MoAB, the conjugate did not bind E6 cells infected with HFRS virus strains. The results
to
are
shown in the table. The results suggest that there may be different serotypes of HFRS virus in China as well as in Sichuan Province. This may assist in the selection of reference strains for producing HFRS vaccine. Health and Anti-epidemic Centre of Sichuan Province,
DONG YOU YAN
Chengdu, Sichuan 610031, People’s Republic of China
CHUAN AN ZHANG
YUN JU XIE
Special Pathogens Branch, Division of Viral Diseases, Center for Infectious Diseases, Centers for Disease Control,
Atlanta, Georgia, USA Health and Anti-epidemic Station of Daxiun Prefecture, Sichuan
J. B. MCCORMICK A. SANCHEZ H. M. ENGELMAN SHANG ZHI CHEN
Health and Anti-epidemic Station of Guang-an County, Sichuan
XIAN SHI GU
Health and Anti-epidemic Station of Da County, Sichuan
WEN TING TANG JIAN ZHANG
PW, Johnson KM. Isolation of the etiologic agent of Korean hemorrhagic fever. J Infect Dis 1978; 137: 298-308. Song G, Hang CS, Liao HX, et al. Antigenjc difference between viral strains causing classical and mild types of epidemic hemorrhagic fever with renal syndrome in China. J Inf Dis 1984; 150: 889-94. Franko MC, Gibbs CJ Jr, Lee PW, Gajdusek DC. Monoclonal antibodies specific for Hantaan virus. Proc Natl Acad Sci USA 1983; 80: 4149-53. van der Groen G, Tkachenko EA, Lvanov AP, Lukashevich I, Provost C, McCormick JB. Antigenic differences between Hantaan and USSR strains of HFRS virus using monoclonal antibodies. In: XIth International Congress for Tropical Medicine and Malaria (Calgary, Canada, 1984): abstr 159.
1. Lee HW, Lee 2.
3.
4.
5. Kitamura T. Present status of HFRS studies in the National Institute of Health Tokyo, Japan. Sendai, Japan: WHO Regional Office for the Western Pacific
WRP/CDS/WG/84.11. 1984: 28-31. 6. Chen BQ, Fu JL, Liao HX, et al. The antigenic analysis of EHF viruses by 35 McAbs. Chin J Microbiol Immunol 1984; 5: 136-39 (English abstract).