Immunocytochemical determination of antigen and epitope specificity of HIV-1-specific B cells in lymph-node biopsies from HIV-1-infected individuals

Laman, Jon D.; Rácz, Paul; Tenner-Rácz, Klara; Klasmeier, Maren; Fasbender, Marianne; Neelen, Conny; Zegers, N.D.; Dietrich, M.; Boersma, W.J.A.; Claassen, Eric

doi:10.1097/00002030-199103000-00002

Cited by 8 publications

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“…Earlier studies suggested that inverse immunohistochemistry might prove useful in localizing HIV-1-specific B cells in histologic specimens (25,26). However, initial attempts to apply this technique to the study of lymph nodes of chronically SHIV-infected rhesus monkeys met with little success.…”

Section: Resultsmentioning

confidence: 99%

“…A number of methodological choices that were made in pursuing this study probably account for the dramatically higher frequency of virus-specific B cells seen in these AIDS virus-infected monkeys than have previously been reported by others (26). First, the evaluated lymph nodes were obtained from a cohort of animals infected with molecularly cloned SHIVs, viruses with defined envelope sequences.…”

Section: Discussionmentioning

confidence: 99%

“…Consequently, recombinant proteins homologous to the infecting virus could be employed as antigen probes. Heterologous probes, as were employed in studies of HIV-infected human lymph nodes (26), detected a substantially smaller fraction of responding B cells. Second, the use of catalyzed signal amplification significantly increased the sensitivity of inverse immunohistochemical staining, allowing detection of Env-binding B cells that would otherwise be overlooked.…”

Section: Discussionmentioning

confidence: 99%

“…This follicular hyperplasia persists throughout an AIDS virus infection until its end stage and is manifested clinically as lymphadenopathy. The lymphadenopathy and follicular hyperplasia are so extreme that a state of nonspecific B-lymphocyte activation has been hypothesized to account for them (26). The present study examines the alternative hypothesis that the B-cell expansion results from a vigorous virus-specific immune response.…”

mentioning

confidence: 94%

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High Frequency of Virus-Specific B Lymphocytes in Germinal Centers of Simian-Human Immunodeficiency Virus-Infected Rhesus Monkeys

Margolin

Saunders

Bronfin

et al. 2002

J Virol

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The etiology of the lymphadenopathy and follicular hyperplasia associated with human immunodeficiency virus type 1 (HIV-1) infection has remained unclear. To determine whether the B-lymphocyte expansions characteristic of this syndrome represent polyclonal and virus-specific processes, the antigen specificity of B cells in lymphoid tissues of monkeys infected with simian-human immunodeficiency virus (SHIV) chimeras was assessed using an inverse immunohistochemical assay with biotinylated HIV-1 envelope gp120 (Env) as an antigen probe. Env-binding B cells were found aggregated in lymph node and splenic germinal centers (GCs). Most Env-binding GCs also contained an unstained population of B cells, suggesting the GCs were formed by a polyclonal (oligoclonal) process. By day 42 following infection, Env-binding B cells were present in 19% of all lymph node GCs. Env-binding cells were present in 25% of GCs even during chronic infection. This extraordinarily high frequency of Env-specific B lymphocytes suggests that the expansion of virus-specific B cells may largely account for the follicular hyperplasia in AIDS virus-infected individuals.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 94%

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High Frequency of Virus-Specific B Lymphocytes in Germinal Centers of Simian-Human Immunodeficiency Virus-Infected Rhesus Monkeys

Margolin

Saunders

Bronfin

et al. 2002

J Virol

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“…They are also of use in immunocytochemical approaches to elucidate the mechanisms of HIV-1-induced immunopathologic developments in situ. We recently described methods to study HIV-1-specific B cells in tissue sections of lymph node biopsy specimens by using antigen-enzyme conjugates (22)(23)(24)(25). In combination with these techniques, HIV-1 variant-specific antibodies applicable in immunocytochemistry would allow study of specific antibody formation in relation to protein expression and antigen localization of the variant under investigation.…”

mentioning

confidence: 99%

Variant-specific monoclonal and group-specific polyclonal human immunodeficiency virus type 1 neutralizing antibodies raised with synthetic peptides from the gp120 third variable domain

Laman¹,

Schellekens²,

Abacıoğlu³

et al. 1992

J Virol

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The third variable (V3) domain of the human immunodeficiency virus type 1 (HIV-1) external membrane glycoprotein gpl20 is of crucial importance in eliciting neutralizing antibodies in infected persons. Polyclonal (PAb) and monoclonal (MAb) antibodies directed against selected epitopes in the V3 domain are valuable tools for analysis of the involvement of such sequences in neutralization and for definition of the relation between amino acid variability and immunological cross-reactions. The aim of this study was to obtain such site-specific antibodies. By using synthetic peptides derived from the V3 domain, a group-specific neutralizing PAb, two high-affinity HIV-1 IIIB neutralizing MAb, and two nonneutralizing MAb were raised. A 15-amino-acid peptide overlapping the tip of the V3 domain of H1V-1 MN was used to produce a rabbit PAb (W0/07). This PAb inhibited syncytium formation induced by HIV-1 IIIB and four field isolates. A similar IllB-derived peptide was used to generate two murine immunoglobulin GI (IgGl) MAb (IIIB-V3-13 and IIIB-V3-34). Pepscan analysis mapped the binding site of IIIB-V3-34 to the sequence IRIQRGPGR. The Kds of IIIB-V3-13 and IIIB-V3-34 for gpl20 were 6.8 x 10-11 and 1.6 x 10-10 M, respectively. These MAb neutralized IIIB but not MN and inhibited syncytium formation induced by IIIB. They are applicable in enzyme-linked immunosorbent assays, immunocytochemistry, and flow cytometry. A peptide covering the left base of the V3 domain was used to generate two murine IgGl MAb (IIIB-V3-21 and IIIB-V3-26). The binding site of HIB-V3-21 was mapped to the sequence INCTRPN. These MAb did not neutralize H1V-1 and did not inhibit syncytium formation. This study supports the notion that HIV-1 neutralizing antibodies suitable for multiassay performance can be obtained with synthetic peptides and that high-affinity MAb can be generated. Such site-specific antibodies are useful reagents in the analysis of HI-1 neutralization. In addition, the crossneutralization of different viral strains by PAb generated through single-peptide immunization is directly relevant to vaccine development.

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Enhanced follicular dendritic cell function in lymph nodes of simian immunodeficiency virus‐infected macaques: Consequences for pathogenesis

Rosenberg¹,

Lewis

Greenhouse

et al. 1997

Eur J Immunol

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HIV and simian immunodeficiency virus (SIV) infections are characterized by several abnormalities in B cell function. Pathogenesis is also associated with marked changes within germinal centers (GC) including hypertrophy and degeneration of follicular dendritic cells (FDC) and accumulation of both viral antigen and activated CD45RO+ CD8+ cells. Since FDC are critical to the generation of antibody-forming cells and specific B cell memory, the simplest assumption is that such B cell defects directly result from virus-induced changes in the GC environment. The present study examined FDC-enriched mesenteric lymph node lymphocyte preparations from early and late stage SIV-infected and uninfected macaques for their ability to support GC reactions in vitro. The results indicate that FDC function as measured by cluster formation, B cell proliferation and SIV-specific antibody production is enhanced in SIV-infected macaques suggesting that, despite FDC atrophy, virus accumulation induces increased FDC-B cell interactions resulting in B cell hyperactivity. The activation and proliferation of CD8+ cells in FDC-enriched cultures further suggest that the infiltrating CD8+ population observed in situ in GC of late-stage SIV/HIV-infected individuals may also benefit from FDC-derived growth signals. Thus, in addition to enhanced B cell proliferation and antibody production, hyperactivity of FDC may potentially promote their own self destruction via the infiltrating CD8+ cells. The increased B cell responsiveness may further exacerbate the disease process due to an overall decrease in the affinity of anti-HIV/SIV antibody, a loss of crucial protective antibodies to other infectious agents and the creation of an environment in which increased trapping of virions facilitates more extensive infection of CD4+ T cells.

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Immunocytochemical determination of antigen and epitope specificity of HIV-1-specific B cells in lymph-node biopsies from HIV-1-infected individuals

Cited by 8 publications

References 0 publications

High Frequency of Virus-Specific B Lymphocytes in Germinal Centers of Simian-Human Immunodeficiency Virus-Infected Rhesus Monkeys

High Frequency of Virus-Specific B Lymphocytes in Germinal Centers of Simian-Human Immunodeficiency Virus-Infected Rhesus Monkeys

Variant-specific monoclonal and group-specific polyclonal human immunodeficiency virus type 1 neutralizing antibodies raised with synthetic peptides from the gp120 third variable domain

Enhanced follicular dendritic cell function in lymph nodes of simian immunodeficiency virus‐infected macaques: Consequences for pathogenesis

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