Scand J Infect Dis 24: 419-421, 1992
HIV-2 Infection in Denmark BIRGIT BAK KVINESDAL’, ANNE MARIE WORM2, BJARNE BRSKOV LINDHARDT3, BIRGITTE L0KKE JENSEN’, CARSTEN MBLLER NIELSEN’ and ANNE-GRETHE POULSEN’
Scand J Infect Dis Downloaded from informahealthcare.com by UB Kiel on 11/06/14 For personal use only.
From the Departments of ’Virology, Statens Seruminstitut, and 2Dermato-Venereology, Bispebjerg Hospital. and ’Laboratory of Tumor Virology, The Fibiger Institute, Copenhagen, Denmark
A collection of 3019 selected serum samples (ss), comprising 329 ss from intravenous drug abusers, 558 ss from homosexual men, 682 samples from persons attending a STD clinic, 100 ss from individuals of African origin, 300 ss from sexual contacts to Africans,QSOss from Danish blood donors who resided in Africa >2 years prior to donating the ss, and 400 ss with equivocal antibody reactions in an HIV-1 Western blot was tested for antibodies against HIV-2 by in-house HIV-2 ELISA and Western blot. Four ss were positive for antibodies against HIV-2. Three of the ss originated from West African men, the fourth belonged to the spouse of one of these men. Three of the samples presented with an uncharacteristic reaction in a HIV-1 Western blot. The study indicates that HIV-2 infection is not yet widespread in Denmark and that it remains closely related to West Africa. B . B . Kvinesdal, MD, Kikhanebakken 63, DK-2840 Holte, Denmark
INTRODUCTION Since the identification of the second “AIDS-causing virus” in 1985 (l),several serological studies from West Africa have revealed HIV-2 to be endemic in several countries ( 2 4 ) . During the last 5 years HIV-2 infection has also been reported from several European countries and from the Americas (5-11). Despite the lack of close historical relation to West Africa as compared to France and Portugal, tourist visits of Danes to West African countries are not uncommon, and further immigration from these countries occurs. Thus we wanted to investigate whether HIV-2 has been introduced into Denmark. MATERIALS AND METHODS Serum samples (ss) A total of 3019 ss from 3019 individuals were tested. Eight different categories were chosen. 329 ss drawn from intravenous drug abusers (IVDA) from an outpatient clinic in Copenhagen, of which 72 were HIV-1 antibody positive; none had AIDS. 200 ss from healthy HIV-1 antibody negative homosexual men from Copenhagen. 358 ss from HIV-1 antibody positive homosexual men from Copenhagen (CDC class I1 and 111). 682 ss from patients attending an outpatient clinic for sexual transmitted diseases (STD) in Copenhagen; 15 were tested HIV-1 antibody positive, none had AIDS. 100 ss from healthy persons originating from Africa. 300 ss from healthy HIV-1 antibody negative Danes who had had sexual contact with an African. 650 ss from healthy Danish blood donors who had been in Africa more than 2 years prior to donating the ss; all of them being HIV-1 antibody negative. 400 ss with inconclusive HIV-1 WB patterns (reactivity against HIV-1 core and pol antigens without reactivity against envelope antigens). Luborutory methods All sera were initially tested for antibodies against HIV-1 using an in-house HIV-1 ELISA (12). Confirmation of the ELISA reactive samples was performed by an in-house HIV-1 WB employing HIV-1 antigen from Dupont (13). Testing of HIV-2 antibodies was done by an in-house HIV-2 ELISA
420 B. B. Kvinesdal et al.
Scand J Infect Dis 24
Table 1. Number and origin of serum samples Individualdpersons
No.
Intravenous drug abusers (IVDA) HIV-1 neg homosexuals HIV-I pos homosexuals STD patients Originating from Africa Having had sex with Africans Danish blood donors having been
329
in Africa
Inconclusive HIV-1 WB
HIV-2 POS
200
358 682 100
300
1 1
650 400
2a
Scand J Infect Dis Downloaded from informahealthcare.com by UB Kiel on 11/06/14 For personal use only.
~
"The samples later turned out to belong to 2 West African men. and WB (14). The HIV-2 WB was considered positive if antibodies against the core protein (p 24) and at least one of the envelope proteins (gp 36 or gp 105/140)were identified.
RESULTS Four of the 3019 ss were found positive for antibodies against HIV-2 (Table I). Three of the positive ss originated from 3 West African men from the Ivory Coast, Guinea Bissau, and Senegal respectively. The fourth ss was from a woman who was married to one of the men. Three of the ss were positive in both the HIV-1 and the HIV-2 ELISA, whereas the fourth ss from the woman was only reactive in the HIV-2 ELISA. For 2 of the 3 ELISA HIV-1 reactive ss, inconclusive WB HIV-1 patterns (reactivity against p24, p55, p68) led to the suspicion of HIV-2 infection. The third ss presented with full reactivity in both HIV-1 WB (reactivity against p24, p31, gp41, p55, p68, gpll0) and HIV-2 WB (reactivity against p24, gp36, p55, p68, gp 105/140). Further analysis using RIPA and immunofluorescence (IFA) revealed that the person was HIV-2 infected (details given in ref. 15). The fourth ss from the woman, presented with p24 reactivity in HIV-1 WB and reactivity against p17, p24, gp36, p55, and gp105/140 in HIV-2 WB. DISCUSSION The overall prevalence of HIV-2 infection in Denmark was expected to be very low. Investigations in other European countries and the USA have revealed that HIV-2 infection is primarily found among immigrants from West Africa and people with sexual relations to West Africans (5,7,11). A few cases have been identified among IVDA's and homosexual men ( 6 9 ) . Further, HIV-2 antibodies have been found in ss presenting with an atypical reactivity in HIV-I WB (5). Our study did not reveal HIV-2 infection among homosexuals, IVDA's, and persons attending a STD clinic. Three of the 4 HIV-2 antibody positive individuals have immigrated from West Africa and the fourth was a spouse of one of the men. In 1988 the prevalence of HIV-1 antibodies among Danish blood donors were found to be 1.3: 100000, a decrease from 2.2: 100000 in 1986 (16). Aproximately 400000 donations are given per year, 40000 as first time donations. Persons with risk behaviour including sexual contact with an African and persons who have been in Africa during the last 2 years are requested to obstain from donating blood. However, we investigated 650 donors who had been in Africa within the last 10 years prior to introducing the exclusion criterias but none of them were HIV-2 antibody positive. The study also points to the difficulties of interpreting WB results as 3 of the 4 HIV-2 antibody positive specimens presented with inconclusive HIV-1 WB patterns. One of the ss presented with reactivities against the core and env proteins in both the HIV-1 WB and the
Scand J Infect Dis 24
HIV-2 infection in Denmark 421
Scand J Infect Dis Downloaded from informahealthcare.com by UB Kiel on 11/06/14 For personal use only.
HIV-2 WB. Only further analysis using other assays identified this sample to be HIV-2 positive (15). Other studies (10) have shown that ss from asymtomatic HIV-2 antibody positive persons would usually react in an HIV-1 ELISA whereas ss from HIV-2 antibody positive AIDS patients usually do not react due to the lack of core antibodies. In this study, a sample originating from a healthy Danish woman married to one of the West African men was not recognized in an HIV-1 ELISA although a strong reactivity against the core protein could be demonstrated in the HIV-2 WB. This indicates that despite core reactivity all HIV-2 antibody positive ss are not positive in an HIV-1 ELISA using whole virus antigen. Thus as the recombinant or peptide-based HIV-1 and HIV-2 ELISA become more specific, the need for performing both assays becomes greater. In conclusion, this study of selected individuals indicates, that HIV-2 infection is not yet widespread in Denmark and that it remains closely related to West Africa. Information of relation to West Africa and inconclusive HIV-1 WB patterns should lead to testing for HIV-2 antibodies,and serological surveillance for HIV-2 infection should be instituted at regular intervals. ACKNOWLEDGEMENT This work was supported by grants from the Danish Medical Research Council (project AIDS 12-7937).
REFERENCES 1. Clavel F, Guetard D, Brun-Vezinet F, Chamaret S, Rey MA, Santos-Ferreia MO, Laurent AG, Daguet C, Katlama C, Rouziux C , Klatzmann D, Champaimaud JL, Montagnier L. Isolation of a new human retrovirus from West African patients with AIDS. Science 233: 343-346, 1986. 2. Denis F, Barin F, Gershy-Damet G, Rey L, Lhuiliier M, Mounier M, Leonard G , Sangare A, Geudeau A, M'Boup S, Essex M, Kanki JP. Prevalence of human T-lymphotropic retrovirus I11 (HIV) and type IV in Ivory Coast. Lancet 1: 40W11. 1987. 3. Poulsen AG, Kvinesdal 8, Aaby P, Mdbak K, Frederiksen K, Dias F, Lauritzen E. Prevalence of and mortality from immunodeficiency virus type 2 in Bissau, West Africa. Lancet 1: 827-830,1989. 4. Odehuri K, De Cock KM, Krebs JW, Moreau J, Rayfield M, McCormick JB, Schochetman G , Bretton R, Bretton G, Ouattara D. HIV-I and HIV-2 infection associated with AIDS in Abidjan, Cote d'lvoire. AIDS 3: 509-512, 1989. 5. Biberfeld G, Bottiger B, Bredberg-Riden U, Putkonen PO, Ericson L, Berglund 0, Starup C, H5kansson C. Findings in four HTLV-IVseropositive women from West Africa. Lancet 2 :1330,1986. 6. Briicker G, Brun-Vezinet F, Rosenheim M . HIV-2 infection in two homosexual men in France. Lancet I: 223. 1987. 7. Courouce AM. HIV-2 in blood donors and in different risk groups in France. Lancet 1: 910,1987. 8. Werner A, Stazewski S, Helms EB,Stille W, Werber K, Kurth R. HIV-2 (West Germany 1984). Lancet 1: 868, 1987. 9. Vanderborght B, De Leys RJ, Nijs P, Van-der-Groen G, Merregaert J, Prinsen H, Van-Heuverswyn H. Isolation of human immunodeficiency virus type 2 from a homosexual man in Belgium. Eur J Clin Microbiol Infect Dis 7: 816-817, 1988. 10. Courouce AM. A prospective study of HIV-2 prevalence in France. AIDS 2: 261-265, 1988. 11. Update: HIV-2 infection, United States 1989. MMWR 38: 572-580, 1990. 12. Nielsen CM. Kvinesdal B, Vestergaard BE Antigen-antibody reaction in solution in capture competition immunoassay for human immunodeficiency virus antibodies. J Clin Microbiol 27: 1609-1612, 1989. 13. Lauritzen E, Lindhardt B 0 . Antibodies against human immunodeficiency virus (HIV) detected by immunoblotting. In: Bjerrum 0, Heegaard NHH, eds. Handbook of immunoblotting of proteins. New York: CRC Press, 119-127, 1988. 14. Kvinesdal BB, Nielsen CM, Poulsen AG, Worm AM, Lindhardt BO, Jensen BL. Performance of human immunodeficiency virus type 2 (HIV-2) Western blot and ELISA: determination of cut-off and cross-reactivity with HIV-1. In preparation. 15. Kvinesdal B, Nielsen CM, Poulsen AG, Hejlyng N. lmmunofluorescence assay for detection of antibodies to human immunodeficieny virus type 2. J Clin Microbiol 27: 2502-2504, 1989. 16. Schmidt K, Dickmeiss E. Screening of .donor blood for anti-HIV. Experience from blood donor screening in 19861988. Ugeskr Lzger 36: 2552-2556, 1990. (In Danish, abstract in English.)
HIV-2 Infection in Denmark BIRGIT BAK KVINESDAL’, ANNE MARIE WORM2, BJARNE BRSKOV LINDHARDT3, BIRGITTE L0KKE JENSEN’, CARSTEN MBLLER NIELSEN’ and ANNE-GRETHE POULSEN’
Scand J Infect Dis Downloaded from informahealthcare.com by UB Kiel on 11/06/14 For personal use only.
From the Departments of ’Virology, Statens Seruminstitut, and 2Dermato-Venereology, Bispebjerg Hospital. and ’Laboratory of Tumor Virology, The Fibiger Institute, Copenhagen, Denmark
A collection of 3019 selected serum samples (ss), comprising 329 ss from intravenous drug abusers, 558 ss from homosexual men, 682 samples from persons attending a STD clinic, 100 ss from individuals of African origin, 300 ss from sexual contacts to Africans,QSOss from Danish blood donors who resided in Africa >2 years prior to donating the ss, and 400 ss with equivocal antibody reactions in an HIV-1 Western blot was tested for antibodies against HIV-2 by in-house HIV-2 ELISA and Western blot. Four ss were positive for antibodies against HIV-2. Three of the ss originated from West African men, the fourth belonged to the spouse of one of these men. Three of the samples presented with an uncharacteristic reaction in a HIV-1 Western blot. The study indicates that HIV-2 infection is not yet widespread in Denmark and that it remains closely related to West Africa. B . B . Kvinesdal, MD, Kikhanebakken 63, DK-2840 Holte, Denmark
INTRODUCTION Since the identification of the second “AIDS-causing virus” in 1985 (l),several serological studies from West Africa have revealed HIV-2 to be endemic in several countries ( 2 4 ) . During the last 5 years HIV-2 infection has also been reported from several European countries and from the Americas (5-11). Despite the lack of close historical relation to West Africa as compared to France and Portugal, tourist visits of Danes to West African countries are not uncommon, and further immigration from these countries occurs. Thus we wanted to investigate whether HIV-2 has been introduced into Denmark. MATERIALS AND METHODS Serum samples (ss) A total of 3019 ss from 3019 individuals were tested. Eight different categories were chosen. 329 ss drawn from intravenous drug abusers (IVDA) from an outpatient clinic in Copenhagen, of which 72 were HIV-1 antibody positive; none had AIDS. 200 ss from healthy HIV-1 antibody negative homosexual men from Copenhagen. 358 ss from HIV-1 antibody positive homosexual men from Copenhagen (CDC class I1 and 111). 682 ss from patients attending an outpatient clinic for sexual transmitted diseases (STD) in Copenhagen; 15 were tested HIV-1 antibody positive, none had AIDS. 100 ss from healthy persons originating from Africa. 300 ss from healthy HIV-1 antibody negative Danes who had had sexual contact with an African. 650 ss from healthy Danish blood donors who had been in Africa more than 2 years prior to donating the ss; all of them being HIV-1 antibody negative. 400 ss with inconclusive HIV-1 WB patterns (reactivity against HIV-1 core and pol antigens without reactivity against envelope antigens). Luborutory methods All sera were initially tested for antibodies against HIV-1 using an in-house HIV-1 ELISA (12). Confirmation of the ELISA reactive samples was performed by an in-house HIV-1 WB employing HIV-1 antigen from Dupont (13). Testing of HIV-2 antibodies was done by an in-house HIV-2 ELISA
420 B. B. Kvinesdal et al.
Scand J Infect Dis 24
Table 1. Number and origin of serum samples Individualdpersons
No.
Intravenous drug abusers (IVDA) HIV-1 neg homosexuals HIV-I pos homosexuals STD patients Originating from Africa Having had sex with Africans Danish blood donors having been
329
in Africa
Inconclusive HIV-1 WB
HIV-2 POS
200
358 682 100
300
1 1
650 400
2a
Scand J Infect Dis Downloaded from informahealthcare.com by UB Kiel on 11/06/14 For personal use only.
~
"The samples later turned out to belong to 2 West African men. and WB (14). The HIV-2 WB was considered positive if antibodies against the core protein (p 24) and at least one of the envelope proteins (gp 36 or gp 105/140)were identified.
RESULTS Four of the 3019 ss were found positive for antibodies against HIV-2 (Table I). Three of the positive ss originated from 3 West African men from the Ivory Coast, Guinea Bissau, and Senegal respectively. The fourth ss was from a woman who was married to one of the men. Three of the ss were positive in both the HIV-1 and the HIV-2 ELISA, whereas the fourth ss from the woman was only reactive in the HIV-2 ELISA. For 2 of the 3 ELISA HIV-1 reactive ss, inconclusive WB HIV-1 patterns (reactivity against p24, p55, p68) led to the suspicion of HIV-2 infection. The third ss presented with full reactivity in both HIV-1 WB (reactivity against p24, p31, gp41, p55, p68, gpll0) and HIV-2 WB (reactivity against p24, gp36, p55, p68, gp 105/140). Further analysis using RIPA and immunofluorescence (IFA) revealed that the person was HIV-2 infected (details given in ref. 15). The fourth ss from the woman, presented with p24 reactivity in HIV-1 WB and reactivity against p17, p24, gp36, p55, and gp105/140 in HIV-2 WB. DISCUSSION The overall prevalence of HIV-2 infection in Denmark was expected to be very low. Investigations in other European countries and the USA have revealed that HIV-2 infection is primarily found among immigrants from West Africa and people with sexual relations to West Africans (5,7,11). A few cases have been identified among IVDA's and homosexual men ( 6 9 ) . Further, HIV-2 antibodies have been found in ss presenting with an atypical reactivity in HIV-I WB (5). Our study did not reveal HIV-2 infection among homosexuals, IVDA's, and persons attending a STD clinic. Three of the 4 HIV-2 antibody positive individuals have immigrated from West Africa and the fourth was a spouse of one of the men. In 1988 the prevalence of HIV-1 antibodies among Danish blood donors were found to be 1.3: 100000, a decrease from 2.2: 100000 in 1986 (16). Aproximately 400000 donations are given per year, 40000 as first time donations. Persons with risk behaviour including sexual contact with an African and persons who have been in Africa during the last 2 years are requested to obstain from donating blood. However, we investigated 650 donors who had been in Africa within the last 10 years prior to introducing the exclusion criterias but none of them were HIV-2 antibody positive. The study also points to the difficulties of interpreting WB results as 3 of the 4 HIV-2 antibody positive specimens presented with inconclusive HIV-1 WB patterns. One of the ss presented with reactivities against the core and env proteins in both the HIV-1 WB and the
Scand J Infect Dis 24
HIV-2 infection in Denmark 421
Scand J Infect Dis Downloaded from informahealthcare.com by UB Kiel on 11/06/14 For personal use only.
HIV-2 WB. Only further analysis using other assays identified this sample to be HIV-2 positive (15). Other studies (10) have shown that ss from asymtomatic HIV-2 antibody positive persons would usually react in an HIV-1 ELISA whereas ss from HIV-2 antibody positive AIDS patients usually do not react due to the lack of core antibodies. In this study, a sample originating from a healthy Danish woman married to one of the West African men was not recognized in an HIV-1 ELISA although a strong reactivity against the core protein could be demonstrated in the HIV-2 WB. This indicates that despite core reactivity all HIV-2 antibody positive ss are not positive in an HIV-1 ELISA using whole virus antigen. Thus as the recombinant or peptide-based HIV-1 and HIV-2 ELISA become more specific, the need for performing both assays becomes greater. In conclusion, this study of selected individuals indicates, that HIV-2 infection is not yet widespread in Denmark and that it remains closely related to West Africa. Information of relation to West Africa and inconclusive HIV-1 WB patterns should lead to testing for HIV-2 antibodies,and serological surveillance for HIV-2 infection should be instituted at regular intervals. ACKNOWLEDGEMENT This work was supported by grants from the Danish Medical Research Council (project AIDS 12-7937).
REFERENCES 1. Clavel F, Guetard D, Brun-Vezinet F, Chamaret S, Rey MA, Santos-Ferreia MO, Laurent AG, Daguet C, Katlama C, Rouziux C , Klatzmann D, Champaimaud JL, Montagnier L. Isolation of a new human retrovirus from West African patients with AIDS. Science 233: 343-346, 1986. 2. Denis F, Barin F, Gershy-Damet G, Rey L, Lhuiliier M, Mounier M, Leonard G , Sangare A, Geudeau A, M'Boup S, Essex M, Kanki JP. Prevalence of human T-lymphotropic retrovirus I11 (HIV) and type IV in Ivory Coast. Lancet 1: 40W11. 1987. 3. Poulsen AG, Kvinesdal 8, Aaby P, Mdbak K, Frederiksen K, Dias F, Lauritzen E. Prevalence of and mortality from immunodeficiency virus type 2 in Bissau, West Africa. Lancet 1: 827-830,1989. 4. Odehuri K, De Cock KM, Krebs JW, Moreau J, Rayfield M, McCormick JB, Schochetman G , Bretton R, Bretton G, Ouattara D. HIV-I and HIV-2 infection associated with AIDS in Abidjan, Cote d'lvoire. AIDS 3: 509-512, 1989. 5. Biberfeld G, Bottiger B, Bredberg-Riden U, Putkonen PO, Ericson L, Berglund 0, Starup C, H5kansson C. Findings in four HTLV-IVseropositive women from West Africa. Lancet 2 :1330,1986. 6. Briicker G, Brun-Vezinet F, Rosenheim M . HIV-2 infection in two homosexual men in France. Lancet I: 223. 1987. 7. Courouce AM. HIV-2 in blood donors and in different risk groups in France. Lancet 1: 910,1987. 8. Werner A, Stazewski S, Helms EB,Stille W, Werber K, Kurth R. HIV-2 (West Germany 1984). Lancet 1: 868, 1987. 9. Vanderborght B, De Leys RJ, Nijs P, Van-der-Groen G, Merregaert J, Prinsen H, Van-Heuverswyn H. Isolation of human immunodeficiency virus type 2 from a homosexual man in Belgium. Eur J Clin Microbiol Infect Dis 7: 816-817, 1988. 10. Courouce AM. A prospective study of HIV-2 prevalence in France. AIDS 2: 261-265, 1988. 11. Update: HIV-2 infection, United States 1989. MMWR 38: 572-580, 1990. 12. Nielsen CM. Kvinesdal B, Vestergaard BE Antigen-antibody reaction in solution in capture competition immunoassay for human immunodeficiency virus antibodies. J Clin Microbiol 27: 1609-1612, 1989. 13. Lauritzen E, Lindhardt B 0 . Antibodies against human immunodeficiency virus (HIV) detected by immunoblotting. In: Bjerrum 0, Heegaard NHH, eds. Handbook of immunoblotting of proteins. New York: CRC Press, 119-127, 1988. 14. Kvinesdal BB, Nielsen CM, Poulsen AG, Worm AM, Lindhardt BO, Jensen BL. Performance of human immunodeficiency virus type 2 (HIV-2) Western blot and ELISA: determination of cut-off and cross-reactivity with HIV-1. In preparation. 15. Kvinesdal B, Nielsen CM, Poulsen AG, Hejlyng N. lmmunofluorescence assay for detection of antibodies to human immunodeficieny virus type 2. J Clin Microbiol 27: 2502-2504, 1989. 16. Schmidt K, Dickmeiss E. Screening of .donor blood for anti-HIV. Experience from blood donor screening in 19861988. Ugeskr Lzger 36: 2552-2556, 1990. (In Danish, abstract in English.)
