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Biomedical subjects

F Barin

Publications and source records attributed to F Barin.

At least 91 records · Page 5Linked to original sources

Site-directed serology using synthetic oligopeptides representing the C-terminus of the external glycoproteins of HIV-1, HIV-2, or SIVmac may distinguish subtypes among primate lentiviruses.

In this study the presence of a highly immunogenic domain located at the C-terminus of the external glycoproteins (EGP) of human and simian immunodeficiency viruses (HIV-1, HIV-2, and SIVMAC) is shown using synthetic oligopeptides as antigens in enzyme immunoassays. This epitope is probably located within the last 13 and 15 residues of the EGP of HIV-1 and HIV-2, respectively. The C terminal epitope of the EGP of SIVMAC may involve residues located more upstream. Among the HIV-2/SIV serotype, we observed that the reactivity to the C-terminal epitope of gp120 was dependent of both species and geographical origin of the samples tested. It seems that this gp 120 C-terminal epitope could distinguish subtypes among the HIV-2/SIV serotype. Further studies, using site-directed enzyme immunoassays with synthetic peptides representing the C-terminus of the EGP derived from a wide variety of HIV2/SIV strains must be performed to confirm this observation. These kinds of assays may constitute important tools for use in seroepidemiological studies and broaden our understanding of the distribution and phylogenetic relationship of primate lentiviruses.

Amino Acid Sequence↗

Characterization of human antibody-binding sites on the external envelope of human immunodeficiency virus type 2.

Antibody-reactive peptide scanning (Pepscan) using overlapping nonapeptides and human sera as probes allows the identification of amino acids contributing significantly to antigen-antibody interaction. Five-hundred and two overlapping nonapeptides derived from the human immunodeficiency virus type 2 strain Rod (HIV-2Rod) external envelope glycoprotein gp125 were synthesized to serve as probes for reactivity with eight sera of HIV-2-infected individuals. Fifteen antibody-binding regions were identified, among which two amino-terminal regions [E3, amino acids (aa) 118 to 132; E4, aa 125 to 141] and four carboxy-terminal regions (E11, aa 303 to 324; E12, aa 340 to 358; E14, aa 436 to 452; E15, aa 486 to 507) were the most antigenic. The amino acids in binding sites E3 and E4 were highly variable among simian immunodeficiency virus (SIV), HIV-2 and HIV-1. The antibody-binding domains E14 and E15 were highly conserved among HIV-2 strains (94% and 86% identity of HIV-2Rod to HIV-2Isy and HIV-2NIHZ, respectively). Both domains had more amino acids in common with SIV (88% for E14, 64% for E15) than with HIV-1 (41% for E14, 45% for E15). Epidemiological studies revealed that the sera of African HIV-2-infected individuals bound the E11 and E15 peptides best (31%, 8/26). The sera of African HIV-1-infected individuals showed significant levels of cross-reactivity to the HIV-2 peptides, especially to the E15 peptide, whereas the sera of European HIV-1-infected individuals showed only moderate levels of cross-reactivity. If peptides covering the E15 epitope were used, African HIV-2-positive sera showed only a low level of cross-reactivity (4%) to E15 of HIV-1 and SIV. African HIV-1-positive sera bound the HIV-1 E15 peptide best (81%, 52/64), but showed high levels of cross-reactivity to SIV E15 (17%, 11/64) and HIV-2 E15 (25%, 16/64). European HIV-1-positive sera showed a high level of reactivity of HIV-1 E15 (91%, 50/55) and a low level of cross-reactivity to the HIV-2 (2%, 1/55), and none to the SIV E15 (0/55) peptide. These results indicate that the immunodominant regions of the HIV-2 external envelope (E11 and E15) align with the most immunodominant regions of HIV-1.

Acquired Immunodeficiency Syndrome↗

Immune response to a major epitope of p24 during infection with human immunodeficiency virus type 1 and implications for diagnosis and prognosis.

A sequential inhibition enzyme-linked immunoassay (SIEIA) using a peroxidase-conjugated monoclonal antibody reacting to the sequence AAEWDRVHP of p24HIV-1 (amino acids 209 to 217 of p55) was developed in order to detect and determine the titer of antibody to this epitope in various populations of human immunodeficiency virus type 1 (HIV-1)-positive patients. There was a good correlation between SIEIA and a commercially available competition assay that uses recombinant p24 protein and polyclonal antibody to HIV-1 antigen, demonstrating the importance of the described epitope. Analysis of sera from French patients showed a decline of antibody to the AAEWDRVHP sequence associated with the progression of AIDS. No decrease was observed with serum samples from African patients. An immune response to the epitope was detected by SIEIA early in the course of seroconversion. Although our SIEIA uses a single p24 epitope, these data are in accordance with previously published studies in which antibodies to the whole p24 were analyzed. Sera reacting to p24 only (indeterminate profiles by Western blot [immunoblot]) did not bind to AAEWDRVHP. This epitope, which is conserved between HIV-1 and HIV-2/simian immunodeficiency virus, appears to be a major antigenic domain of p24. The area containing the sequence AAEWDRVHP and the corresponding monoclonal antibody may serve as a convenient alternative to whole purified p24 and polyclonal antibody in diagnostic and prognostic assays.

Amino Acid Sequence↗

Fine serotyping of human immunodeficiency virus serotype 1 (HIV-1) and HIV-2 infections by using synthetic oligopeptides representing an immunodominant domain of HIV-1 and HIV-2/simian immunodeficiency virus.

In this study, enzyme immunoassays for detection of type-specific antibodies to human immunodeficiency viruses (HIV) were developed by using short peptides corresponding to sequences located within the immunodominant domain of the transmembrane glycoproteins of both HIV-1 and HIV-2-simian immunodeficiency virus (SIV). The assays were highly sensitive with currently available sera from various geographical areas. Furthermore, they appeared to be more specific in HIV serotyping than the Western blot (immunoblot) assay, since all of the sera were clearly discriminated as one or the other type. It was also shown that in contrast to HIV-1, the C-terminal cysteine residue (amino acid 620, SIV from captive macaques, Mm142 strain) of the HIV-2-SIV peptide is not necessary for recognition of the peptide by antibody to HIV-2.

Amino Acid Sequence↗

[Virologic diagnosis of HIV infection in the pregnant woman and newborn infant].

The virological diagnosis of HIV infection in pregnant women mainly consists of detecting antibodies to structural viral antigens. The difficulty lies more in the interpretation of the confirmatory assay than in technical problems. The result is critical since the continuation or interruption of pregnancy is dependent upon it. The diagnosis of HIV infection in newborns has been hampered by the presence of passively acquired maternal antibodies. The diagnosis at birth therefore must include culture of peripheral blood mononuclear cells for virus isolation and detection of serum antigen. However, due to the difficulty or lack of sensitivity of these tests new diagnostic procedures, such as the polymerase chain reaction, must be developed to improve the accuracy of the early diagnosis. Such new approaches must be evaluated very carefully before being introduced in routine diagnosis.

AIDS Serodiagnosis↗

Competitive enzyme-immunoassays using native viral antigens to discriminate between HIV-1 and HIV-2 infections.

Human immunodeficiency virus (HIV) strains can be separated into two serotypes: HIV-1 and HIV-2. In the study reported herein, we developed and evaluated competitive enzyme-immunoassays (CEIA-1 for the detection of antibody to HIV-1, CEIA-2 for the detection of antibody to HIV-2) to discriminate serologically between HIV-1 and HIV-2 infections. In most of the cases, the serotyping of known reactive serum samples was done easily with the CEIAs, showing a similar specificity to the Western blot. Such competitive assays could represent alternative procedures for serotyping HIV infection and would limit the need for expensive Western blots, especially in economically poor countries.

Antibodies, Viral↗

Confirmatory tests for human T-cell leukemia virus type 1 (HTLV-1): western blot compared with RIPA on African sera.

Confirmation of human T-Cell leukemia virus type 1 (HTLV-1) seropositivity calls for reactivity against at least 2 proteins encoded by 2 different genes, revealed by Western blot (WB) and/or radioimmuno-precipitation assay (RIPA). To evaluate the use of WB as a basis for applying these criteria, we conducted a study of two types of WB and compared them with RIPA patterns. The first part of the work, performed with 40 African sera, used Dupont de Nemours commercialized WB and an 'in-house' WB. Both WB detected antibody to proteins encoded by 2 different genes: antibody to gag products were revealed equally by both WB, but commercialized WB detected antibody to tax protein whereas the 'in-house' WB detected antibody to env protein (gp46) more efficiently. The second part of the work, conducted with 158 African sera, compared results of an 'in-house' virus lysate WB and RIPA. Our data show a perfect concordance between the two procedures when sera were clearly positive by WB (gag + env reaction). Sera reacting to p19 and p24 (both gag) by WB were confirmed positive by RIPA in 75% of the cases. The majority of the indeterminate WB profiles not confirmed by RIPA presented isolated gag reactivity (p15 or p19 or p24).

Blotting, Western↗

Linear B-cell epitopes of the major core protein of human immunodeficiency virus types 1 and 2.

Nine murine monoclonal antibodies directed to the major core protein p24 of human immunodeficiency virus type 1 (HIV-1) were obtained and then tested by using an epitope mapping system (Pepscan) covering the whole p24HIV1 protein to characterize antigenic domains. Four different linear epitopes were identified. Monoclonal antibodies recognizing three of these epitopes also reacted to p26HIV2 in Western blotting (immunoblotting). A monoclonal antibody specific for the fourth epitope, located at position 179 to 188 of the gag polyprotein p55HIV1 (human T-cell lymphotropic virus type 3B strain), did not react with HIV type 2 (HIV-2) core proteins. The corresponding sequence is constant in all known HIV-2 and simian immunodeficiency virus (SIV) isolates, including a very divergent SIV strain from African green monkeys (SIVagm/tyo). This observation may be relevant to the phylogeny of primate lentiviruses. Two of the conserved epitopes might be immunogenic during natural infection and could therefore be used for diagnosis and prognosis purposes. These two epitopes are AAEWDRVHP and EIYKRWII, starting at positions 209 and 260 of the polyprotein p55HIV1, respectively.

Amino Acid Sequence↗

HIV 2 infection among blood donors and other subjects in France. The "Retrovirus" Study Group of the French Society of Blood Transfusion.

During a 3 year period, from August 1985 to August 1988, 18 HIV 2-infected blood donors were detected in France as a result of systematic HIV 1 screening. These sera were characterized as HIV 2 by specific Western blot and synthetic peptides. Within the same period, 40 other HIV 2 infected subjects were identified by our study group, independently of blood donations. Thirty of these 58 subjects living in France originate from West Africa (8 blood donors), 3 (I blood donor) are Portuguese men who had lived in West Africa and 25 (9 blood donors) are of French origin; among these, 16 have had a known close contact with West Africa. When HIV 2-infected subjects were asymptomatic, cross reactivity between antibodies to HIV 2 and HIV 1 proteins was generally observed with the two-step ELISA assays using total HIV 1 proteins; it was poor with the competitive assays and variable with the assays using recombinant proteins. When the subjects had signs of immunodeficiency, cross reactivity decreased. The data confirm that HIV 2 is not widespread in France and that most of HIV 2-infected but asymptomatic subjects are recognized by several HIV 1 ELISA assays. Accordingly, systematic screening for HIV 2 with an additional test cannot be recommended at the present time, but a combined HIV 1 and HIV 2 test will be useful when available.

Acquired Immunodeficiency Syndrome↗

Serum capacity to inhibit reverse transcriptase in vitro distinguishes HIV-1 infection from HIV-2 or SIV infection.

The inhibition of HIV-1 and SIV reverse transcriptase by human and rhesus macaque serum positive for HIV-1 or HIV-2/SIV antibodies was studied. The domain to which reverse transcriptase-inhibiting antibodies were elicited appeared to be highly antigenic. A total of 67% (48 of 72) of individuals had HIV-1 reverse transcriptase-inhibiting (RTI) antibodies 1 year after seroconversion for HIV-1, 90% (9 of 10) of HIV-2 antibody positive persons had SIV RTI antibodies, and all four experimentally SIV-infected rhesus macaques developed SIV RTI antibodies. Low cross-reactivity between HIV-1 and HIV-2/SIV RTI antibodies was observed. Of 10 HIV-1 RTI sera, 2 reduced SIV RT activity by more than 50% (mean reduction 85 versus 24%). Only 1 of 9 SIV RTI human sera reduced HIV-1 RT (mean reduction 74 versus 25%). This serum, however, showed a shared reactivity against both HIV-1 and HIV-2. These results indicate that the HIV-1 domain inducing RTI antibodies is antigenically different from the HIV-2/SIV domain.

Acquired Immunodeficiency Syndrome↗

Prevalence of antibody to human T cell leukemia virus type 1 (HTLV-1) in populations of Ivory Coast, West Africa.

A large cross-sectional serologic survey for human T cell leukemia virus type 1 (HTLV-1) antibody was conducted in 3,177 Ivory Coast residents to evaluate the prevalence of HTLV-1 and to determine possible risk factors and correlates of HTLV-1 infection. Of the 3,177 serum samples, 110 (3.5%) were positive for antibody to HTLV-1 by indirect immunofluorescence assay and Western blot. The prevalence of HTLV-1 antibody in the general adult population was 1.8% and increased significantly with age. No difference between males (1.5%) and females (2%) was found. The highest prevalences were observed in female prostitutes (7.4%), patients with neurologic syndromes (5.8%), and lepers (13.7%). The high prevalence of HTLV-1 infection in prostitutes suggests that heterosexual contact is involved in the transmission of HTLV-1 and that prostitutes could play an important role in the spread of the virus in Africa. The high prevalence of HTLV-1 in patients with neurologic syndromes confirms the association between HTLV-1 and some type of neuropathies, as has been observed in the West Indies and Japan. The high prevalence observed in lepers deserves further investigation to find the cause of the association. Twenty-five individuals, including prostitutes, were coinfected with HTLV-1 and human immunodeficiency virus (HIV). Prospective studies are necessary to evaluate the exact role of HTLV-1 alone or in combination with HIV in inducing specific diseases.

Child↗