Mubarak Osei-Kwasi

Isolation and characterization of HIV-2 from an AIDS patient in Ghana.

This report describes the isolation and characterization of a retrovirus of the HIV-2 group from a Ghanaian AIDS patient which has different restriction patterns from previously reported HIV-2 viruses. The virus was morphologically very similar to HIV-1 and HIV-2, and had Mg2+-dependent reverse transcriptase. Like previous HIV isolates, it induced severe cytopathic effects in CD4-positive human lymphoid cell lines. Its major proteins were shown to be gp110, p66, p55, p41, gp32, p30 and p26 by Western blot analysis. In dot-blot hybridization experiments, the virus hybridized with a HIV-2 DNA probe, but not with HIV-1 and SIVagm probes in stringent conditions. These data indicate that this Ghanaian virus is a HIV-2 group virus. However, in a Southern blot hybridization experiment, the restriction patterns of this virus, designated HIV-2 [GH-1], were quite different from those of previously reported HIV-2 viruses from West Africa isolated at the Pasteur Institute.

SEROLOGICAL SURVEY OF HIV-1, HIV-2 AND HUMAN T-CELL LEUKEMIA VIRUS TYPE 1 FOR SUSPECTED AIDS CASES IN GHANA

Objective:To determine seroprevalence among suspected AIDS patients in Ghana in relation to clinical manifestations. Materials and methods:Blood samples and medical records were collected from 290 Ghanaian patients with suspected AIDS in 1990 and 1992. Seroprevalence of HIV-1, HIV-2 and human T-cell leukemia virus (HTLV-1) were investigated by the particle agglutination method, indirect immunofluorescence assay, the monoepitope enzyme-linked immunosorbent assay and Western blot. Results:The specimens were classified into five serologic categories: 78 were HIV-1-positive (26.9%), 25 were HIV-2-positive (8.6%), 17 dual-positive (5.9%), 16 indeterminate (5.5%) and 154 seronegative (53.1%). No significant difference was found between the clinical symptoms of patients with HIV-1 and HIV-2 infection. Of the patients, 14 (4.8%) were HTLV-1-seropositive, of whom 11 were also HIV-positive, indicating a significant correlation between the two groups of viral infections (P<0.01). However, there was no evidence of an increase in severity of symptoms in cases of dual infection with HTLV-1 and HIV. Conclusions:HIV-1 infection is now dominant in Ghana in contrast to our previous survey in 1986 which showed the dominance of HIV-2. The change in seroprevalence suggests that an HIV-1 epidemic has been developing in recent years in this country, where HIV-2 was originally endemic. A relatively high prevalence of dual-reactive specimens implies the existence of highly cross-reactive strains of HIV or frequent coinfection with HIV-1 and HIV-2 in the region. The large number of seronegative patients with clinically diagnosed AIDS raises the question of the inadequacy of AIDS definitions based on clinical manifestations only.

SURFACE MARKER PATTERNS OF T CELLS AND EXPRESSION OF INTERLEUKIN-2 RECEPTOR IN MEASLES INFECTION

The surface marker patterns of T cells of Ghanaian children during measles infection were studied and an attempt was made to demonstrate T cell activation and viability in vitro after activation in vivo by measles virus. The frequencies of CD4+ and CD8+ naive T cells in measles patients were high while their memory T cells were remarkably reduced with no sign of proliferation even at the acute phase of the illness. The reduction of memory T cells was prolonged during the convalescent phase (2 months after onset). The anti‐CD3 monoclonal antibody‐induced expression of interleukin‐2 receptor α chain (IL‐2R/CD25) was significantly suppressed; however, the addition of phorbol 12‐myristate 13‐acetate or ionomycin caused a remarkable recovery of CD25 expression. On simple culture, an appreciable proportion of T cells from measles patients died rapidly in contrast with only a few T cells from healthy controls doing so. The suppression of CD25 expression was still demonstrated during the convalescent phase of the disease. Taken together these results suggest unresponsiveness and activation‐induced cell death of T cells during severe measles infection in Ghanaian children. Furthermore the prolonged abnormalities of T cells (i.e. decreased memory T cells and inhibition of CD25 expression during the convalescent phase) might be related to post‐measles infection immunosuppressive status.

Antibody response to 17D yellow fever vaccine in Ghanaian infants

OBJECTIVES To assess the seroresponses to yellow fever vaccination at 6 and 9 months of age; assess any possible adverse effects of immunization with the 17D yellow fever vaccine in infants, particularly at 6 months of age. METHODS Four hundred and twenty infants who had completed BCG, OPV and DPT immunizations were randomized to receive yellow fever immunization at either 6 or 9 months. A single dose of 0.5 ml of the reconstituted vaccine was administered to each infant by subcutaneous injection. To determine the yellow fever antibody levels of the infants, each donated 1 ml whole blood prior to immunization and 3 months post-immunization. Each serum sample was titred on Vero cells against the vaccine virus. FINDINGS The most common adverse reactions reported were fever, cough, diarrhoea and mild reactions at the inoculation site. The incidences of adverse reactions were not statistically different in both groups. None of the pre-immunization sera in both age groups had detectable yellow fever antibodies. Infants immunized at 6 months recorded seroconversion of 98.6% and those immunized at 9 months recorded 98% seroconversion. The GMT of their antibodies were 158.5 and 129.8, respectively. CONCLUSIONS The results indicate that seroresponses to yellow fever immunization at 6 and 9 months as determined by seroconversion and GMTs of antibodies are similar. The findings of good seroresponses at 6 months without significant adverse effects would suggest that the 17D yellow fever vaccine could be recommended for use in children at 6 months in outbreak situations or in high risk endemic areas.

Hospital-Based Surveillance for Viral Hemorrhagic Fevers and Hepatitides in Ghana

Background Viral hemorrhagic fevers (VHF) are acute diseases associated with bleeding, organ failure, and shock. VHF may hardly be distinguished clinically from other diseases in the African hospital, including viral hepatitis. This study was conducted to determine if VHF and viral hepatitis contribute to hospital morbidity in the Central and Northern parts of Ghana. Methodology/Principal Findings From 2009 to 2011, blood samples of 258 patients with VHF symptoms were collected at 18 hospitals in Ashanti, Brong-Ahafo, Northern, Upper West, and Upper East regions. Patients were tested by PCR for Lassa, Rift Valley, Crimean-Congo, Ebola/Marburg, and yellow fever viruses; hepatitis A (HAV), B (HBV), C (HCV), and E (HEV) viruses; and by ELISA for serological hepatitis markers. None of the patients tested positive for VHF. However, 21 (8.1%) showed anti-HBc IgM plus HBV DNA and/or HBsAg; 37 (14%) showed HBsAg and HBV DNA without anti-HBc IgM; 26 (10%) showed anti-HAV IgM and/or HAV RNA; and 20 (7.8%) were HCV RNA-positive. None was positive for HEV RNA or anti-HEV IgM plus IgG. Viral genotypes were determined as HAV-IB, HBV-A and E, and HCV-1, 2, and 4. Conclusions/Significance VHFs do not cause significant hospital morbidity in the study area. However, the incidence of acute hepatitis A and B, and hepatitis B and C with active virus replication is high. These infections may mimic VHF and need to be considered if VHF is suspected. The data may help decision makers to allocate resources and focus surveillance systems on the diseases of relevance in Ghana.

Immunological reactivities of Ghanaian sera with HIV-1 HIV-2 and simian immunodeficiency virus SIVagm.

To learn more about the relative prevalence of viruses from the human immunodeficiency virus (HIV)/simian immunodeficiency virus (SIV) groups in Ghana, serum samples were collected in 1986 from 47 men and women with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC), 57 apparently healthy individuals, and 11 AIDS-free hospital inpatients. Western blot analysis revealed a total of 46 reactive sera. 43 of the 47 serum samples from those with AIDS or ARC were positive; 6 were seropositive for HIV-1, 18 for HIV-2, 17 for both HIV-1 and HIV-2, and the remaining 2 were not reactive with glycoproteins. Of the 2/10 individuals with mild symptoms of HIV infection who proved to be seropositive, 1 was positive for HIV-2 and 1 for HIV-1 and HIV-2. There were no seropositive reactions among the AIDS-free hospital patients, and only 1 such reaction (seropositive for HIV-2) among the healthy individuals. These findings indicate that both HIV-1 (6 cases in this series) and HIV-2 (20 cases) are responsible for the development of AIDS in Ghana, and that there is a high prevalence (18 cases) of cross-reactivity between the 2 viruses. There was no evidence of SIV infection. Further research is needed to determine whether these findings are a result of cross-reactivity between envelope proteins of HIV-1 and HIV-2, double infection of AIDS patients, or infection with a new variant strain. Since prostitutes comprised 25 of the 47 AIDS/ARC patients and 6 of the 10 with mild symptoms of HIV infection, they are an important target for preventive efforts.

Clinically diagnosed AIDS cases without evident association with HIV type 1 and 2 infections in Ghana

Case studies increasingly point to the potential of HIV-negative individuals with clinically diagnosed AIDS. The results of hematologic tests on blood samples collected from 227 Ghanaian people with AIDS suggest the potential existence of many such cases. Blood samples were subjected to ELISA particle agglutination indirect fluorescent assay western blot and mono-epitope ELISA tests. 48 individuals tested positive for only HIV-1 17 for only HIV-2 11 were dual positive 16 indeterminate and 135 seronegative. All of those testing seronegative for HIV nonetheless suffered weight loss prolonged diarrhea and chronic fever. Many also exhibited lymphadenopathy tuberculosis dermatological diseases and neurological disorders. While many of these individuals may have been misdiagnosed as having AIDS the existence of unknown agents which are antigenically different from any know type of HIV should be seriously considered.

The Middle to 3′ End of the Hiv-1 Vif Gene Sequence is Important for Vif Biological Activity and Could be Used for Antisense Oligonucleotide Targets

The human immunodeficiency virus type-1 (HIV-1)-encoded Vif protein is essential for viral replication, virion production, and pathogenicity. HIV-1 Vif interacts with the endogenous human APOBEC3G protein (an mRNA editor) in target cells to prevent its encapsidation into virions. Some studies have established targets within the HIV-1 vif gene that are important for its biologic function; however, it is important to determine effective therapeutic targets in vif because of its critical role in HIV-1 infectivity and pathogenicity. The present study demonstrates that virions generated in transfected HeLa-CD4+ cells, especially from HIV-1 vif frame-shift mutant (3′-Δvif; 5561-5849), were affected in splicing and had low infectivity in MT-4 cells. In addition, HIV-1 vif antisense RNA fragments constructed within the same region, notably the region spanning nucleic acid positions 5561-5705 (M-3′-AS), which corresponds to amino acid residues 96–144, significantly inhibited HIV-1 replication in MT-4 and reduced the HIV-1 vif mRNA transcripts and reporter gene (EGFP) expression. The generated virions showed low secondary infection in H9 cells. These data therefore suggest that the middle to the 3′ end of vif is important for its biological activity in the target cells.

REACTIVITIES OF ANTIBODIES TO HIV AND SIV IN HUMAN SERA IN KENYA, GABON, AND GHANA

The isolation of simian immunodeficiency virus (SIV) from African non-human primates and 2 new human retroviruses from people in West Africa (HIV-2 and HTLV-IV) which are antigenically closely related to SIV indicate the importance of viral ecological studies on HIV/SIV especially in Africa. This is a report on reactivity with the antigens of SIV derived from African green monkey and HIV-1 of antibodies in 29 anti-HIV-positive human sera from East and West Africa. SIV shares antigenicity with the gag gene products of HIV-1 but not with its envelope gene protein products. Of the 29 sera 12 were from patients with AIDS or AIDS-related complex (ARC) in Kenya 5 from apparently healthy people in Gabon and 12 from prostitutes manifesting ARC in Ghana. From the serological findings it is conceivable that the antibody-positive people in Kenya are infected with a virus that is antigernically similar to prototype HIV (HIV-1) while those in Gabon and Ghana are infected with a virus that is closely related to SIV such as GTLV-IV or or HIV-2. To test this idea isolates from these seropositive persons were obtained in Kenya and Ghana. Analysis of viral proteins and genomes revealed that the Kenyan isolates were indistinguishable from the prototype HIV-1 group whereas the Ghanaian isolate belongs to the HIV-2 virus group.