- Email: [email protected]
Surveillance for HIV-1 subtypes O and M in Kenya
doses of corticosteroids in treating refractory rejection of renal allograft/ Only after rescue with MMF did our young patient return to normal renal function. She is presently well controlled on cyclosporin (300 mg/m2 daily) and MMF (375 mg/day), and has maintained normal renal function and a growth spurt ever since hospital discharge. At this institution all paediatric renal transplant recipients are now started on antithymocyte globulin, cyclosporin, and MMF. This regimen may eliminate the risk of breakthrough rejection episodes, which can arise with a steroid-free immunosuppressive regimen. This is a definite advantage in view of the risk of steroid toxicity in children.
raises concern that the number of with HIV may increase. We thank Guido
facilities to confirm our findings, and the director of KEMRI for permission to publish these results. This project was funded by KEMRI HIV Diagnostic, account no 4.467.000.020.
EM Songok, D K Libondo, M C Rotich, S A Kenya Medical Research Institute, Nairobi, Kenya 1
2
4
Oogo, P M Tukei
Heyndrickx L, Alary M, Janssens W, Davo N, van der Groen G. HIV1 group O and group M dual infection in Bénin. Lancet 1996; 347: Songok EM, Tukei PM, Mulaa FJ. Serological investigation of HIV-1 subtype strains in transmission in Nairobi. East Afr Med J
variant 3
3
Groen and staff at the Institute of Tropical
Medicine, Antwerp, Belgium, for technical guidance and provision of
Department of Nephrology and Tissue Culture Laboratory, Odense University Hospital, DK-5000 Odense C, Denmark
2
of double infection
902-03.
S A Birkeland
1
van
cases
Birkeland SA, Elbirk A, Rohr N, Jørgensen KA. CyA monotherapy in children-long-term effect. Transplant Proc 1994; 26: 2610-16. European Mycophenolate Mofetil Cooperative Study Group. Placebocontrolled study of mycophenolate mofetil combined with cyclosporin and corticosteroids for prevention of acute rejection. Lancet 1995; 345: 1321-25. Sollinger HW, for the US Renal Transplant Mycophenolate Mofetil Study Group. Transplantation 1995; 60: 225-32. Sollinger HW, Deierhoi MH, Belzer FO, Diethelm AG, Kauffman RS. RS-61443: a phase I clinical trial and pilot rescue study. Transplantation 1992; 53: 428-32.
4
5
1996; 73: 57-59. Peeters M, Nkengasong J, Willems B, et al. Antibodies to V3 loop peptides derived from chimpanzee lentiviruses and the divergent ANT70 isolate in human sera from different geographic regions. AIDS 1994; 8: 1657-61. Janssens W, Fransen K, Loussert Ajaka I, et al. Diagnosis of HIV-1 group O infection by polymerase chain reaction. Lancet 1995; 346: 451-52. Nkengasong JN, Peeters M, van den Haesevelde M, et al. Antigenic evidence of presence of aberrant HIV-1 ANT70 virus in Cameroon and Gabon. AIDS 1993; 7: 1536-38.
No HHV8 in non-Kaposi’s sarcoma mucocutaneous lesions from immunodeficient
Surveillance for HIV-1 subtypes O and M in
HIV-positive patients
Kenya SiR-Heyndrickx and colleagues (March 30, p 902)’ report HIV infection in a Benin woman with a unique dual HIV subtype M and subtype 0 infection. We report here a similar reactivity in an HIV-positive sample collected in Nairobi during ongoing efforts to determine the HIV subtypes in circulation in Kenya and their association with infectivity and pathogenesis. These efforts require collection of HIV-positive serum samples and peripheral blood mononuclear cells from various populations and the subtyping of virus by strainspecific peptide enzyme-linked immunosorbent assay (ELISA).’ Between September, 1995, and February, 1996, we investigated patients for HIV 1 subtypes in Nairobi, Kisumu, Kakamega, Eldoret, and Nakuru towns. We used three types of subtype-O-specific V3 loop peptides, ANT70, VI686, and CA9.3 Serum samples reacting to any of the peptides with an ELISA optical density cutoff value of 0-8 and above were judged positive. To save on cost, only samples with a cutoff value of 2-0 and above were confirmed by a subtype-0specific line immunoassay (LIA, Innogenetics, Belgium),4 and those so reactive were then tested by PCR with primers specific for subtype M and subtype 0/ 22 (7-7%) of these samples were subtype O-positive in at least one peptide (ELISA cutoff OD 0-8). 13 of the 22 were reactive with all peptides. The highest reactivity was recorded with VI686 peptide (20/22). On confirmation by LIA, one of four samples with ELISA ODs of 2-0 and above reacted with group 0 ANT70 V3-loop peptide, which is regarded as confirmation for the presence of group 0.2,5 The
same
LIA-
positive sample gave a positive PCR amplification on use of both subtype M and subtype 0 primers. This sample was collected in Nairobi from an adult Kenyan male who had tested HIV-positive on routine medical checkup. He had no history of travel outside Kenya and, at the time of sample collection, he
was without symptoms. This is the first indication of HIV subtype 0 in Kenya. The finding of a dual subtype M and subtype 0 infection
1700
SiR-HHV8-DNA has been detected in different forms of Kaposi’s sarcoma (KS),’ and it has been suggested that it plays a part in causing this tumour. HHV8 has been also found in body-cavity lymphomas,2 in Castleman’s tumours,3 and in skin lesions of four HIV-negative organ-transplant patients with neither lymphoma nor KS.4 This has been interpreted as indicating a widespread distribution of HHV8 in the general population,which would suggest it may not be the cause of KS. We looked for HHV8-DNA sequences in non-KS mucocutaneous lesions of HIV-infected and non-infected patients. We used conventional and nested PCR to amplify HHV8-DNA fragments from biopsy specimens. The quality of DNA samples and the presence of PCR-inhibitor factors were controlled for by amplifying j3-actin sequences, and only positive samples were considered in this study. The sensitivity of PCR was determined by serial dilution of genomic DNA from KS tissue. Our protocol was sensitive enough to detect HHV8-DNA in 10 ng KS-derived genomic DNA by conventional PCR and in 1 ng by nested PCR.
*Molluscum contagiosum (2), viral warts (4), melanoma (1), granuloma annulare (1), seborrhoeic keratosis (1), necrotic ulcer (1), hairy leukoplakia (2), pilar cyst (1), penanal ulcer (3), arthropod reaction (1), oesophagus ulcer (1), erythema (1), psonasis (2), herpes zoster (2), abscess (1), dermatitis (2), prurigo nodulans (1), herpes simplex virus infection (1). tLelomyoma (3), melanoma (8), basal cell carcinoma (11), pityriasis rosea (4), molluscum contagiosum (6), psoriasis vulgaris (6), viral wart (8), pseudolymphoma
(7). Table : Detection of HHVS-DNA in KS and non-KS skin lesions of HIV-1 seropositive and seronegative patients
raises concern that the number of with HIV may increase. We thank Guido
facilities to confirm our findings, and the director of KEMRI for permission to publish these results. This project was funded by KEMRI HIV Diagnostic, account no 4.467.000.020.
EM Songok, D K Libondo, M C Rotich, S A Kenya Medical Research Institute, Nairobi, Kenya 1
2
4
Oogo, P M Tukei
Heyndrickx L, Alary M, Janssens W, Davo N, van der Groen G. HIV1 group O and group M dual infection in Bénin. Lancet 1996; 347: Songok EM, Tukei PM, Mulaa FJ. Serological investigation of HIV-1 subtype strains in transmission in Nairobi. East Afr Med J
variant 3
3
Groen and staff at the Institute of Tropical
Medicine, Antwerp, Belgium, for technical guidance and provision of
Department of Nephrology and Tissue Culture Laboratory, Odense University Hospital, DK-5000 Odense C, Denmark
2
of double infection
902-03.
S A Birkeland
1
van
cases
Birkeland SA, Elbirk A, Rohr N, Jørgensen KA. CyA monotherapy in children-long-term effect. Transplant Proc 1994; 26: 2610-16. European Mycophenolate Mofetil Cooperative Study Group. Placebocontrolled study of mycophenolate mofetil combined with cyclosporin and corticosteroids for prevention of acute rejection. Lancet 1995; 345: 1321-25. Sollinger HW, for the US Renal Transplant Mycophenolate Mofetil Study Group. Transplantation 1995; 60: 225-32. Sollinger HW, Deierhoi MH, Belzer FO, Diethelm AG, Kauffman RS. RS-61443: a phase I clinical trial and pilot rescue study. Transplantation 1992; 53: 428-32.
4
5
1996; 73: 57-59. Peeters M, Nkengasong J, Willems B, et al. Antibodies to V3 loop peptides derived from chimpanzee lentiviruses and the divergent ANT70 isolate in human sera from different geographic regions. AIDS 1994; 8: 1657-61. Janssens W, Fransen K, Loussert Ajaka I, et al. Diagnosis of HIV-1 group O infection by polymerase chain reaction. Lancet 1995; 346: 451-52. Nkengasong JN, Peeters M, van den Haesevelde M, et al. Antigenic evidence of presence of aberrant HIV-1 ANT70 virus in Cameroon and Gabon. AIDS 1993; 7: 1536-38.
No HHV8 in non-Kaposi’s sarcoma mucocutaneous lesions from immunodeficient
Surveillance for HIV-1 subtypes O and M in
HIV-positive patients
Kenya SiR-Heyndrickx and colleagues (March 30, p 902)’ report HIV infection in a Benin woman with a unique dual HIV subtype M and subtype 0 infection. We report here a similar reactivity in an HIV-positive sample collected in Nairobi during ongoing efforts to determine the HIV subtypes in circulation in Kenya and their association with infectivity and pathogenesis. These efforts require collection of HIV-positive serum samples and peripheral blood mononuclear cells from various populations and the subtyping of virus by strainspecific peptide enzyme-linked immunosorbent assay (ELISA).’ Between September, 1995, and February, 1996, we investigated patients for HIV 1 subtypes in Nairobi, Kisumu, Kakamega, Eldoret, and Nakuru towns. We used three types of subtype-O-specific V3 loop peptides, ANT70, VI686, and CA9.3 Serum samples reacting to any of the peptides with an ELISA optical density cutoff value of 0-8 and above were judged positive. To save on cost, only samples with a cutoff value of 2-0 and above were confirmed by a subtype-0specific line immunoassay (LIA, Innogenetics, Belgium),4 and those so reactive were then tested by PCR with primers specific for subtype M and subtype 0/ 22 (7-7%) of these samples were subtype O-positive in at least one peptide (ELISA cutoff OD 0-8). 13 of the 22 were reactive with all peptides. The highest reactivity was recorded with VI686 peptide (20/22). On confirmation by LIA, one of four samples with ELISA ODs of 2-0 and above reacted with group 0 ANT70 V3-loop peptide, which is regarded as confirmation for the presence of group 0.2,5 The
same
LIA-
positive sample gave a positive PCR amplification on use of both subtype M and subtype 0 primers. This sample was collected in Nairobi from an adult Kenyan male who had tested HIV-positive on routine medical checkup. He had no history of travel outside Kenya and, at the time of sample collection, he
was without symptoms. This is the first indication of HIV subtype 0 in Kenya. The finding of a dual subtype M and subtype 0 infection
1700
SiR-HHV8-DNA has been detected in different forms of Kaposi’s sarcoma (KS),’ and it has been suggested that it plays a part in causing this tumour. HHV8 has been also found in body-cavity lymphomas,2 in Castleman’s tumours,3 and in skin lesions of four HIV-negative organ-transplant patients with neither lymphoma nor KS.4 This has been interpreted as indicating a widespread distribution of HHV8 in the general population,which would suggest it may not be the cause of KS. We looked for HHV8-DNA sequences in non-KS mucocutaneous lesions of HIV-infected and non-infected patients. We used conventional and nested PCR to amplify HHV8-DNA fragments from biopsy specimens. The quality of DNA samples and the presence of PCR-inhibitor factors were controlled for by amplifying j3-actin sequences, and only positive samples were considered in this study. The sensitivity of PCR was determined by serial dilution of genomic DNA from KS tissue. Our protocol was sensitive enough to detect HHV8-DNA in 10 ng KS-derived genomic DNA by conventional PCR and in 1 ng by nested PCR.
*Molluscum contagiosum (2), viral warts (4), melanoma (1), granuloma annulare (1), seborrhoeic keratosis (1), necrotic ulcer (1), hairy leukoplakia (2), pilar cyst (1), penanal ulcer (3), arthropod reaction (1), oesophagus ulcer (1), erythema (1), psonasis (2), herpes zoster (2), abscess (1), dermatitis (2), prurigo nodulans (1), herpes simplex virus infection (1). tLelomyoma (3), melanoma (8), basal cell carcinoma (11), pityriasis rosea (4), molluscum contagiosum (6), psoriasis vulgaris (6), viral wart (8), pseudolymphoma
(7). Table : Detection of HHVS-DNA in KS and non-KS skin lesions of HIV-1 seropositive and seronegative patients