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The objective of this study was to investigate the presence of non-subtype B HIV-1 in Denmark. The C2-V3-C3 region of the env gene from proviral DNA obtained from patients suspected of being infected with non-subtype B virus was PCR-amplified and directly sequenced. The DNA sequences were aligned with full-length HIV-1 reference strains from each subtype and analysed using the phylogenetic package PHYLIP 3.1. The neighbour-joining method was used with 100 bootstraps. Of the 144 patients included in this study C2-V3-C3 sequences were obtained from 129 patients (90%). The phylogenetic analyses showed that virus from 49 patients (38%) was subtype A, 39 (30%) subtype C, 9 (7%) subtype D, 14 (11%) subtype CRF01_AE, 16 (12%) subtype B, 1 (1%) subtype F and 1 (1%) subtype J. This study demonstrates that almost all subtypes can be detected in Denmark; all non-subtype B infections could be traced to countries with a high prevalence of non-subtype B virus.

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BACKGROUND: commercial HIV-1 qualitative DNA PCR tests have the potential to detect virus in patients in whom antibody tests may be ineffective, such as patients with primary HIV infection and infants born to HIV seropositive mothers. However, the genetic diversity of HIV-1 raises concern about the ability of the PCR tests to detect all current subtypes. OBJECTIVES: to asses the sensitivity of the Amplicor HIV-1 test on 126 whole-blood samples representing seven different subtypes and to investigate the sensitivity when the standard assay was modified by including the primer pair SK145 and SKCC1B. RESULTS: of the 126 HIV-1 infected persons, 113 were tested positive and 13 were DNA PCR negative. On the basis of these results, the standard Amplicor HIV-1 test had a sensitivity of 90% in our cohort. In addition, 9% of the positive samples showed a low reactivity but above the cut-off of the assay. The standard assay yielded sensitivities of 100% for subtype B (n=16), D (n=9) and G (n=1), but only 83% for subtype A (n=41), 98% for subtype C (n=43), 79% for subtype E (n=14) and 0% for subtype F (n=2). All samples with low reactivity were non-B subtype. Eight of the DNA PCR negative samples, four subtype A, one C and three E were amplified with the modified Amplicor HIV-1 test with addition of SK145/SKCC1B primers. Using this modified protocol, six samples out of eight became positive. However, two samples (one A and one C) remained DNA PCR negative. CONCLUSION: this study confirms that the Amplicor HIV-1 test does not detect all subtypes with equivalent sensitivity and 10% of the samples, tested negative. Thus, it is preferable to add the SK145/SKCC1B primers to the standard test, where infection with non-B subtype is suspected.

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A case of fatal infection with Strongyloides stercoralis in a previously healthy young African with no known immunosuppression is reported. The patient suffered severe gastrointestinal bleeding and despite intensive treatment died of multiorgan failure. Postmortem, signs of ulcerating T-cell lymphoma were found in a jejunal specimen, and the patient was found to be infected with HTLV-1. Gastrointestinal bleeding in relation to infection with Strongyloides stercoralis is discussed as well as possible relations between the severity of infection, lymphoma and HTLV-1.

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The study describes the use of postexposure antiretroviral prophylaxis (PEP) after occupational exposure to HIV in Denmark in the period 1/1 1997-31/12 1998. Forty-seven accidentally exposed persons received PEP in this period, of whom 14 were nurses, 13 physicians and 11 were not employed in the health care system. In 23 cases side effects to PEP were described. In 18 cases the index person was i.v. drug abuser and in 11 cases homosexual/bisexual men. In 18 cases the HIV status of the index person was unknown at the time of exposure. Ten of these index persons were subsequently tested and all found HIV-negative. It is concluded, that the frequency of occupational exposure to HIV is unacceptably high. Further it is emphasised that in cases where the HIV status of the index person is unknown, PEP should only be instituted if the index person is at risk of being HIV infected and has signs of HIV infection.

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Pneumococcal infections are frequently observed in patients with human immunodeficiency virus (HIV) infection and active immunization has been recommended as prophylaxis in this patient group. We studied 103 out-patients with asymptomatic or mildly symptomatic HIV infection with respect to specific IgG and IgG2 pneumococcal antibodies before and after vaccination with a 23-valent pneumococcal polysaccharide vaccine. A significant increase ( > 2-fold) in IgG and IgG2 antibody levels was observed after 1 month in 69/103 patients (67%) with no correlation with the CD4 cell count at the time of vaccination. The response rate was not influenced by concurrent treatment with anti-retroviral monotherapy, or by age or gender. After immunization a strong correlation between IgG and IgG2 anti-pneumococcal antibodies was demonstrated. Nevertheless, 12 months after vaccination the specific antibody titres were not significantly different from pre-vaccination values. In conclusion, antibodies induced by pneumococcal vaccination in patients with HIV infection have a short duration. This raises the question as to whether vaccination will have any impact on clinical end-point in this group of patients.

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In 1990-91, 237 female sex workers from Copenhagen were enrolled in a larger study performed in 9 European countries. None of 206 women accepting serological testing was HIV-infected despite the fact that 36 (17.5%) were injecting drug users (IDU). Whereas 95% of the women always used condoms with clients over the last 6 months, this proportion was 25% and 9% respectively for casual and regular non-paying partners (P < 0.0001 for both comparisons). Those reporting at least one STD over the last year had more non-paying sexual partners than the others (P < 0.01) and the frequency of STD was lower in women who always used condoms with non-paying partners (7% vs 31%, P = 0.01). Women working on the street were more often IDU than others (78% vs 7%, P < 0.001). Independently of drug use, street prostitutes also tended to have more clients (P = 0.007) and more STD (P = 0.05). The striking differences in condom use with clients as compared to non-paying partners and the association between STD and sexual behaviours with such partners but not with clients show that specific interventions should be designed to promote safer sex with non-paying partners.

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In a community study, the HIV-1 and HIV-2 antibody status of the inhabitants of 100 randomly chosen houses in Bissau, West Africa, were followed from 1987 to 1989. There was no HIV-1 infection alone, while the HIV-2 seroprevalence in adults was 8.9% (58 of 652) in 1987 and 10.1% (61 of 603) in 1989. HIV-2 seroprevalence in 15- to 39-year-olds was 6.1% in 1987 and 11.3% in newcomers in 1989 [the Mantel-Haentzel weighted relative risk (RRMH) = 1.86; 95% confidence interval (CI): 1.07-3.24]. Three hundred thirty adults who were HIV-2 seronegative in 1987 were reexamined in 1989; seven had seroconverted. Follow-up time was 700 person years, giving an incidence of HIV-2 infection of 1 per 100 person years. With a history of sexually transmitted disease (STD), the RR of seroconverting was 9.95 (2.31-42.80). Blood transfusions received since 1987 did not result in seroconversions. No case of vertical transmission of HIV-2 was seen. There was an excess mortality in those who were HIV-2 seropositive; however, it was statistically significant only for children (RR = 22.27; 95% CI: 6.92-71.70; p < 0.0001), not for adults (RR = 2.16; 95% CI: 0.81-5.76; p = 0.23), when considering death from disease only.

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In order to evaluate the role of sexual transmission and parenteral transmission of hepatitis C virus (HCV) in homosexual men and intravenous drug users (IVDU) serum samples from 147 homosexual men and 126 IVDU were tested for anti-HCV. Anti-HCV was found in two (1.4%) of the homosexual men and in 123 (98%) of IVDU. The presence of anti-HCV could not be correlated to the presence of HBV markers or HIV-antibodies. HCV is widespread among Danish IVDU. Risk of sexual transmission seems low even though sexual contact is a much more prevalent risk factor than needle sharing.

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Markers of hepatitis B virus (HBV) infection were measured in 465 non-drug-abusing heterosexually transmitted disease (STD) patients. HBV markers were found in altogether 70 persons, of whom 7 were HBsAg carriers. Those chronically infected were all born in HBV intermediate/high endemic areas. Gonorrhoea was the only STD that was correlated to an increased risk of HBV markers. Number of sexual partners, sex and age was not correlated with HBV infection, irrespective of country of origin. The risk of having HBV markers in an STD clientele in Copenhagen was highly dependent on the country of birth, as the prevalences were 7% (21/307) in persons born in Denmark, 19% (9/47) in those born in other low endemic areas and 36% (40/111) in those born in intermediate/high endemic areas. Falling incidence of gonorrhoea and other STD may render it difficult to point out risk factors indicative of HBV immunization in heterosexual STD patients. In low-risk countries, screening for HBV markers should however be offered to all immigrants and refugees as a part of an HBV immunization program.

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A total of 40 multiple sclerosis (MS) patients from Denmark and 10 from the Faroes were examined for antibodies with affinity to human T cell leukemia/lymphoma virus type 1 (HTLV-I) and human immunodeficiency virus type 1 and 2 (HIV-1 and HIV-2). Using ELISA, MS patients and a group of healthy controls did not differ significantly in their reactivities to HTLV-I. However, elevated reactivities were recorded with 5 MS sera, whereas only 2 of the sera from the controls produced highly values. Ten patients with other neurological diseases all seemed to exhibit low reactivity in HTLV-I ELISA. The reactivities of 2 MS sera decreased considerably by absorption with an HTLV-I lysate. In immunofluorescence assay, two other MS sera reacted with HTLV-I transformed cell lines as well as with non-infected cells. Examined by Western blotting (WB), a single MS serum produced a distinct HTLV-I p19 band. With ELISA for detection of HIV-1 and HIV-2 antibodies, 2 MS sera exhibited borderline reactions. Further examination of these two sera by WB revealed weak reactivities against p24 and p53 of HIV-1. One the whole, the present observations do not suggest that a putative MS retrovirus would be closely related with HTLV-I, HIV-1 or HIV-2.

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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, 650 ss from Danish blood donors who resided in Africa greater than 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.

PIP: Clinicians drew blood samples from 3019 people living in Denmark to determine whether HIV-2 had reached that country. The groups tested included drug users from Copenhagen, healthy HIV-1 positive and negative homosexual men from Copenhagen, patients at a clinic for sexually transmitted diseases (STDs), healthy Africans, Danes who had sexual intercourse with an African, Danish blood donors who went to Africa more than 2 years before they gave a blood sample, and people who had inconclusive HIV-1 Western Blot (WB) patterns. Laboratory personnel used an in-house HIV-1 ELISA and an in-house HIV-2 ELISA to test all samples and an in-house HIV-2 test. 4 (.13%) samples tested positive for HIV-2. 3 of the serum samples were from men from the Ivory Coast, Guinea Bissau and Senegal. The 4th sample belonged to the wife of one of these men. She was positive only for HIV-2 while the 3 men also tested positive for HIV-1. The serum of 2 of the 3 people who tested ELISA HIV-1 reactive had inconclusive HIV-1 WB patterns who tested ELISA HIV-1 reactive had inconclusive HIV-1 WB patterns which made the researchers suspect HIV-2 infection. The woman's serum reacted to the core and env proteins in both the HIV-1 Wb and HIV-2 WB, the HIV-1 ELISA was negative. RIPA and immunofluorescence tests confirmed HIV-2 infection. Her case demonstrates the need to do both HIV-1 and HIV-2 ELISA tests. None of the 650 blood donors who had been in Africa within the last 10 years tested positive for HIV-2. These findings indicated that HIV-2 was not prevalent in Denmark and was limited to West Africa. Health workers whose patients have ties with West Africa and have an inconclusive HIV-1 WB pattern should request testing for HIV-2. The researchers suggested that serological surveillance for HIV-2 should be done at regular intervals.

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Twenty-nine human immunodeficiency virus type 2 (HIV-2) seropositive women identified in a cross-sectional study in Bissau in 1987 participated in a follow-up study in 1988, where each was matched for age and marital status with two HIV-2 seronegative women. Detailed information about all pregnancies was obtained. The HIV-2 seropositive women and their controls had similar mean numbers of pregnancies, live children, children who died, and abortions. The HIV-2 seropositive women did not have a greater risk of having had an abortion or a child who died than did the HIV-2 seronegative women. No difference in survival was seen between children born to HIV-2 seropositive and HIV-2 seronegative women. The H/S-ratios and CD4 numbers were lower in the seropositive group, but none had values lower than 0.4 and 0.4 x 10(9)/L, respectively. Seven prospectively observed children born to HIV-2 seropositive mothers showed no sign of infection. The lack of evidence of transmission of HIV-2 from mother to child is suggested to be due to the absence of marked immunodeficiency in this random sample of the general population.

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A novel competition ELISA for detection of antibodies against HIV-1 was developed. The assay is based on competition at the single epitope level and utilises a human monoclonal antibody and an E. coli-produced fragment of the transmembrane glycoprotein gp41. The sensitivity of the assay was 100% in tests on 247 serum samples obtained from 219 individuals previously shown to be HIV-1 antibody positive by both conventional indirect ELISA and the immunoblotting test. The patients represented various clinical and immunological stages of HIV-1 infection. Likewise, the specificity of the assay was 100% in tests on 105 serum samples from normal individuals previously tested negative by indirect ELISA. Further, among 105 serum samples selected due to consistent false positive reactions in the indirect ELISA only 2 samples (1.9%) demonstrated false positive reactions in the competition ELISA, i.e. 98.1% specificity. Finally, only 2 of 57 (3.5%) serum samples from HIV-2 infected individuals showed positive reactions in the assay, while 54 (94.7%) had absorbance values similar to the negative controls. These results demonstrate that human monoclonal antibodies may form the basis for highly sensitive and specific assays for detection of antibodies to HIV-1.

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In a community study in Guinea-Bissau, West Africa, 47 HIV-2-seropositive cases and 87 matched controls were evaluated immunologically using immuno-alkaline phosphatase linked to avidin-biotin complex for the assessment of CD4 and CD8 status. HIV-2-seropositive individuals had significantly lower total numbers of CD4 cells and CD4/CD8 ratios, 38% having a total number of CD4 cells less than or equal to 0.5 x 10(9)/l and 36% having a CD4/CD8 ratio less than or equal to 0.8. Total numbers of CD4 cells less than or equal to 0.5 x 10(9)/l or CD4/CD8 ratio less than or equal to 0.8 were found in 53% of the HIV-2 seropositives compared with 11% among controls [odds ratio (OR) = 7.3; 95% confidence interval (CI): 3.1-17.1]. Lymphadenopathy was significantly more frequent among HIV-2 seropositives than among controls (OR = 3.4; 95% Cl: 1.5-7.6). HIV-2 seropositives with lymphadenopathy had significantly fewer lymphocytes (P = 0.008) and lower total CD4 (P = 0.029) and total CD8 number (P = 0.011) than HIV-2 seropositives without lymphadenopathy. This study indicates that HIV-2 has a significant immunosuppressive effect.

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A total of 115 women were found to be HIV-antibody positive during the period January 1985 to September 1988 in the three large hospitals in Copenhagen. Thirty-five women (31%) had been infected heterosexually, 63 (55%) were intravenous drug addicts, 12 (10%) had been infected by transfusion and in five (4%) the mode of transmission was unknown. In 1988, 73% of the women who were found to be HIV-positive were infected heterosexually while only 14% were infected in this manner in 1985. Out of the heterosexually HIV-infected women, 25 (71%) had probably been infected by a man from a risk group while nine further women were infected by a known HIV-positive man with-out connection to any risk group. Eleven women (31%) stated that they practised "safe sex" prior to the HIV test while 22 (74%) did so after the positive HIV-test, which represents a considerable improvement. Out of the drug addicted women, six (10%) stated that they had always practiced "safe sex" prior to the test while 13 (28%) did so after the HIV-test. Approximately 1/3 of these women periodically shared syringes and needles with others. It is concluded that heterosexual contact among HIV-infected women in Copenhagen is a route of transmission of significance. Women infected heterosexually appear to alter their sexual behavior when they are made aware of the positive HIV-test.

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In order to describe the risk pattern including sexual behaviour among HIV-infected women in Copenhagen we studied the charts of all women tested seropositive between January 1985 and August 1988 in the three main hospitals handling HIV/AIDS. One hundred and fifteen women were positive for HIV antibodies: 35 (31%) were infected by heterosexual contact and 63 (55%) were intravenous drug users. Among the heterosexually transmitted cases 25 (71%) had intercourse with a man from a high risk group and nine women had intercourse with a known HIV antibody positive man without known risk factors. Use of condoms was claimed by 31% of the heterosexually-transmitted women before the HIV test, and by 74% (P = 0.002) after the test. In 1988 73% of the women tested HIV positive were infected by heterosexual contact, in 1985 only 14%. We conclude that among HIV infected women in Copenhagen heterosexual transmission is a problem. These women seem to change sexual behaviour when informed about HIV positivity.

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The prevalence of HIV-infection was studied in a district hospital in Botswana, in southern Africa. Sera from 466 patients were analysed. The sexually transmitted diseases (STD) group consisted of 175 women and 178 men, who consulted the STD-clinic for complaints which could be attributed to STD or infertility and the antenatal clinic (ANC) group consisted of 113 pregnant women, who attended routine ANC. Sera were analysed with HIV-ELISA, and 3% were positive. All sera were negative in Western blot and were considered false-positive in HIV-ELISA. In the STD group, 42% of the women and 39% of the men, were seropositive for syphilis, while 41% were positive in the ANC group. The figures for clinically demonstrated genital lesions were 33%, 60% and 13%, respectively. It is concluded that HIV-infection is a new disease in Botswana, and that the prevalence is still low in the rural population examined in the present survey.

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