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1.
Article in English | MEDLINE | ID: mdl-9377122

ABSTRACT

The purpose of this study was to document which genetic subtypes of HIV-2 are present in Guinea-Bissau and to investigate whether asymptomatic HIV-2 carriers and AIDS patients carry distinct genetic variants. A secondary aim was to correlate proviral DNA load to clinical and immunologic status of the patients. Thirty-eight asymptomatic HIV-2 carriers and 11 AIDS patients from Bissau, Guinea-Bissau were included in a cross-sectional study in which HIV-2 env V3 sequences, HIV-2 DNA load, and CD4-positive (CD4+) lymphocyte counts were determined. Phylogenetic analyses showed that all investigated subjects carried subtype A HIV-2 variants and that the sequences from AIDS patients and asymptomatic carriers did not form distinct subclusters in the tree. As expected, patients with AIDS had significantly higher median HIV-2 DNA load than did asymptomatic carriers (4.6 vs. 2.0 log10 HIV-2 DNA copies/10(6) CD4+ lymphocytes). Our study indicates that the HIV-2 epidemic in Guinea-Bissau is almost exclusively caused by subtype A HIV-2 variants and that the HIV-2 infections among the asymptomatic carriers and AIDS cases included in the study do not have distinct epidemiologic histories.


PIP: HIV-2 is associated with AIDS but is less pathogenic than HIV-1. HIV-2 is endemic in West Africa, with the highest prevalence in Guinea-Bissau; epidemiologic studies in the country have found HIV-2 seroprevalence in the general population to be 8-10%. HIV-2 seroprevalence increases with age and peaks near age 50-59 years, although the mean age of HIV-2-associated AIDS cases is near 40 years. Only scattered cases of HIV-2 have been reported outside of West Africa, with some concentration in Portugal, France, and India. 38 asymptomatic HIV-2 carriers and 11 AIDS patients from Bissau were included in a cross-sectional study in which HIV-2 env V3 sequences, HIV-2 DNA load, and CD4-positive lymphocyte counts were determined. Phylogenetic analyses showed that all investigated subjects carried subtype A HIV-2 variants and that the sequences from AIDS patients and asymptomatic carriers did not form distinct subclusters in the tree. Subjects with AIDS had significantly higher median HIV-2 DNA load than did asymptomatic carriers.


Subject(s)
Acquired Immunodeficiency Syndrome/virology , DNA, Viral/analysis , Genetic Variation/genetics , HIV-2/genetics , Viral Load , Acquired Immunodeficiency Syndrome/epidemiology , Acquired Immunodeficiency Syndrome/immunology , Adolescent , Adult , Aged , Amino Acid Sequence , CD4 Lymphocyte Count , DNA Primers/chemistry , Enzyme-Linked Immunosorbent Assay , Female , Guinea-Bissau/epidemiology , HIV Envelope Protein gp160/genetics , Humans , Male , Middle Aged , Molecular Sequence Data , Polymerase Chain Reaction
2.
Proc Natl Acad Sci U S A ; 93(20): 10864-9, 1996 Oct 01.
Article in English | MEDLINE | ID: mdl-8855273

ABSTRACT

Phylogenetic analyses are increasingly used in attempts to clarify transmission patterns of human immunodeficiency virus type 1 (HIV-1), but there is a continuing discussion about their validity because convergent evolution and transmission of minor HIV variants may obscure epidemiological patterns. Here we have studied a unique HIV-1 transmission cluster consisting of nine infected individuals, for whom the time and direction of each virus transmission was exactly known. Most of the transmissions occurred between 1981 and 1983, and a total of 13 blood samples were obtained approximately 2-12 years later. The p17 gag and env V3 regions of the HIV-1 genome were directly sequenced from uncultured lymphocytes. A true phylogenetic tree was constructed based on the knowledge about when the transmissions had occurred and when the samples were obtained. This complex, known HIV-1 transmission history was compared with reconstructed molecular trees, which were calculated from the DNA sequences by several commonly used phylogenetic inference methods [Fitch-Margoliash, neighbor-joining, minimum-evolution, maximum-likelihood, maximum-parsimony, unweighted pair group method using arithmetic averages (UPGMA), and a Fitch-Margoliash method assuming a molecular clock (KITSCH)]. A majority of the reconstructed trees were good estimates of the true phylogeny; 12 of 13 taxa were correctly positioned in the most accurate trees. The choice of gene fragment was found to be more important than the choice of phylogenetic method and substitution model. However, methods that are sensitive to unequal rates of change performed more poorly (such as UPGMA and KITSCH, which assume a constant molecular clock). The rapidly evolving V3 fragment gave better reconstructions than p17, but a combined data set of both p17 and V3 performed best. The accuracy of the phylogenetic methods justifies their use in HIV-1 research and argues against convergent evolution and selective transmission of certain virus variants.


Subject(s)
Genes, env , Genes, gag , HIV Infections/transmission , HIV-1/genetics , Phylogeny , RNA, Viral/genetics , Amino Acid Sequence , Base Sequence , Female , Humans , Male , Methods , Molecular Sequence Data
3.
Rev Med Chil ; 123(10): 1189-98, 1995 Oct.
Article in Spanish | MEDLINE | ID: mdl-8733309

ABSTRACT

We studied the evolution of HIV-1 infection and immune response during six years in two twins born from an infected mother. The children had a continuous progression of the infection, proved by CD4+ cell count, serum anti-HIV antibodies, cultivable virus and proviral load. Now, both children are on antiviral treatment. The analysis of serum antibodies showed a different immune response in both children. One of them developed higher levels of antibodies directed against viral proteins and synthetic peptides derived from their aminoacid sequence. In this child, the amount of cultivable virus increased less than in his twin. Nucleotide sequencing of a part of viral genoma, showed that the virus belonged to the B subtype, prevalent in America and Europe. The observed differences in viral sequences suggest a different selective pressure in both twins. This phenomenon could be related to the observed differences in immune response.


Subject(s)
Acquired Immunodeficiency Syndrome/immunology , Diseases in Twins/etiology , HIV Antibodies/blood , Acquired Immunodeficiency Syndrome/blood , Acquired Immunodeficiency Syndrome/therapy , Acquired Immunodeficiency Syndrome/transmission , Adult , Amino Acid Sequence , Base Sequence , CD4 Lymphocyte Count , Child, Preschool , Diseases in Twins/genetics , Female , Follow-Up Studies , Genome, Viral , HIV Envelope Protein gp120/physiology , HIV Seropositivity/immunology , HIV-1/genetics , HIV-1/isolation & purification , Humans , Infectious Disease Transmission, Vertical , Male , Molecular Sequence Data , Time Factors
4.
Virology ; 209(1): 136-46, 1995 May 10.
Article in English | MEDLINE | ID: mdl-7747463

ABSTRACT

HIV-1 can be subdivided into at least nine genetic subtypes (A through H and O), but in Europe and the United States there is an almost complete dominance of subtype B. In this study three Swedish HIV-1 transmission chains of subtypes other than subtype B have been biologically and molecularly characterized. The three index cases were African men. The p17 gag and env V3 regions of the HIV-1 genome were directly sequenced from uncultured lymphocytes. Phylogenetic analyses showed that the HIV-1 variants with each transmission group were genetically closely related, supporting the epidemiological information. The individuals in transmission groups I (n = 3) and II (n = 2) carried subtype G and D virus, respectively. Interestingly, all four individuals in transmission group III displayed a recombinant genotype with subtype D p17 gag sequence and subtype A V3 sequence. The biological phenotype of virus isolates (rapid/high, syncytium-inducing; or slow/low, non-syncytium-inducing) correlated with the clinical stage of the infected individual. The study also suggested that the correlation between biological phenotype and V3 genotype that has been established for subtype B HIV-1 variants may be valid also for other subtypes. This study demonstrates that HIV-1 variants of subtypes other than B, including a subtype A/D recombinant, are being transmitted in Europe.


Subject(s)
HIV Infections/transmission , HIV Infections/virology , HIV-1/classification , HIV-1/genetics , Viral Proteins , Adult , Amino Acid Sequence , Female , Gene Products, gag/genetics , Gene Products, pol/genetics , Genes, env , Genes, gag , Genes, pol , Genetic Variation , Genotype , HIV Antigens/genetics , HIV Envelope Protein gp120/genetics , HIV Infections/epidemiology , Humans , Infant , Male , Molecular Sequence Data , Peptide Fragments/genetics , Phenotype , Pregnancy , Pregnancy Complications, Infectious/virology , Recombination, Genetic , Sequence Homology, Amino Acid , Sweden/epidemiology , gag Gene Products, Human Immunodeficiency Virus
5.
J Virol ; 68(9): 5918-24, 1994 Sep.
Article in English | MEDLINE | ID: mdl-7520096

ABSTRACT

Transmission of human immunodeficiency virus type 1 (HIV-1) from a male accused of rape and deliberate transmission of HIV-1 was investigated by sequencing of the HIV-1 pol and gag genes from virus obtained from the male and from the female victim. Parts of the reverse transcriptase and p17gag genes were amplified and directly sequenced from uncultured peripheral blood mononuclear cells. The sequences were compared with sequences from 21 unrelated HIV-1-infected controls from the same geographic area (Stockholm, Sweden). Bootstrap analysis of phylogenetic trees demonstrated that the sequences from the female were significantly more closely related to the sequences from the male than to sequences from the controls. Furthermore, we found that the male and female shared two distinct genetic variants of HIV-1. In p17gag the major variant had an unusual, out-of-frame deletion of 3 nucleotides which the minor variant lacked. These results indicated that the male had transmitted more than one infectious unit to the female. From this study we concluded that it was highly likely that the HIV-1 strains carried by the male and female were closely epidemiologically linked.


Subject(s)
Genes, gag , Genes, pol , HIV-1/genetics , Rape , Base Sequence , DNA Primers/chemistry , Drug Resistance, Microbial , Female , HIV Reverse Transcriptase , Humans , Male , Molecular Sequence Data , Phylogeny , RNA-Directed DNA Polymerase/chemistry , Sequence Alignment , Sequence Homology, Amino Acid , Zidovudine/pharmacology
7.
J Clin Microbiol ; 30(4): 801-5, 1992 Apr.
Article in English | MEDLINE | ID: mdl-1572966

ABSTRACT

Serum samples with indeterminate Western blot (WB) tests from 61 individuals whose sera were positive by enzyme-linked immunosorbent assay (ELISA) were studied in order to characterize their putative reactions with the human immunodeficiency virus (HIV) proteins and to resolve the HIV infection status of these individuals. The reaction observed by WB could not be confirmed either by radioimmunoprecipitation assay and subsequent electrophoresis (RIPA) or by use of LiaTek (Organon Teknika, Turnbout, The Netherlands) in 28% of the samples. Of the 86 samples that were indeterminate by WB, 66 reacted with p24 by WB; this reaction was confirmed by RIPA in only 21 (32%) and by LiaTek in 49 (74%) of the 66 samples. On the other hand, none of the indeterminate samples that reacted with HIV envelope proteins by WB did so by LiaTek, while 50% precipitated at least some of these proteins in the RIPA. The sensitivities of the three methods for detecting the antibody reaction with the different HIV proteins, which were studied with serial dilutions of positive serum samples, were similar. Thus, a lower sensitivity of RIPA or LiaTek does not seem to be the cause for the lack of reaction of the WB-indeterminate samples by these two methods. Sequential samples from individuals whose serum samples reacted by the three methods gave reproducible results, but all showed low antibody titers. Peripheral blood mononuclear cells obtained from three of the four individuals with sequential samples that reacted with HIV env proteins by WB and RIPA were negative for HIV provirus DNA after amplification by the polymerase chain reaction.


Subject(s)
Blotting, Western , HIV Antibodies/blood , HIV Infections/diagnosis , HIV-1/immunology , DNA, Viral/genetics , HIV Infections/immunology , HIV Infections/microbiology , HIV Seropositivity/diagnosis , HIV Seropositivity/immunology , HIV Seropositivity/microbiology , HIV-1/genetics , HIV-1/isolation & purification , Humans , Immunoassay , Leukocytes, Mononuclear/microbiology , Polymerase Chain Reaction , Radioimmunoprecipitation Assay
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