Molecular Profile of an Antibody Response to HIV-1 as Probed by Combinatorial Libraries

Barbas, Carlos F.; Collet, Thomas A.; Amberg, Willi; Roben, Paul; Binley, James Μ.; Hoekstra, Denise; Cababa, Doug; Jones, Teresa M.; Williamson, R. Anthony; Pilkington, Glenn; Haigwood, Nancy L.; Cabezas, Edelmira; Satterthwait, Arnold C.; Sanz, Iñaki; Burton, Dennis R.

doi:10.1006/jmbi.1993.1203

Cited by 221 publications

(140 citation statements)

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“…Previous work with IgG phage display libraries generated from bone marrow of patients has shown that high affinity Abs with a given specificity are only isolated from relatively small immune libraries similar to that generated here, if the donor has reasonable numbers of specific plasma/B cells and corresponding serum Abs with the given specificity. Similar Abs were not isolated from seronegative donor libraries, demonstrating the importance of the immunization process (24,46,47). In addition, competition between the selected phage display Abs and the serum of the donor verified cloning of the major specificities of the serum response (48).…”

Section: Discussionmentioning

confidence: 92%

Translocation of an Intracellular Antigen to the Surface of Medullary Breast Cancer Cells Early in Apoptosis Allows for an Antigen-Driven Antibody Response Elicited by Tumor-Infiltrating B Cells

Hansen

Nielsen

Ditzel

2002

The Journal of Immunology

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Tumor-infiltrating lymphoplasmacytic cells are a key feature of medullary carcinoma of the breast (MCB), a distinct subtype of human breast cancer that, despite cytologically anaplastic characteristics, has a more favorable prognosis than other types of breast cancer. Since it has been proposed that the improved clinical outcome is due at least in part to the presence of a prominent lymphoplasmacytic cell infiltrate in the tumor stroma, we recently examined the tumor-infiltrating B cell response in MCB and showed that it is oligoclonal and directed against an intracellular protein translocated to the cell surface upon MCB cell apoptosis. Human Abs cloned from MCB lymphoplasmacytic infiltrate-derived phage display libraries and reflecting the dominant part of the response were used to identify the target Ag as actin. Here, we have characterized in detail the cloned human IgG Abs and the translocation process of actin to the cell surface of apoptotic MCB cells. Our analysis shows that the cloned Abs bind specifically and with high affinity to actin, as determined by ELISA and surface plasmon resonance. Sequence analysis revealed that the Abs are highly somatically mutated, with high replacement to silent ratios, indicative of an Ag-driven, affinity-matured response. Interestingly, the tumor-infiltrating B cells in half the MCB patients mainly exhibited an IgG2 response, while IgG1 dominated in the others. To gain insight to the molecular events that may elicit such an Ab response, we examined the translocation of actin to the cell surface of apoptotic MCB cells using flow cytometry and laser scanning cytometry. Our results show that actin becomes exposed on the cell surface of a large proportion of apoptotic MCB cells as an early apoptotic event. We propose that the Ab response against actin produced by tumor-infiltrating B lymphoplasmacytic cells is Ag-driven, affinity-matured, and elicited due to the increased rate of apoptosis occurring within the MCB tumor that facilitates the translocation and proteolytic fragmentation of intracellular proteins.

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Section: Discussionmentioning

confidence: 92%

Translocation of an Intracellular Antigen to the Surface of Medullary Breast Cancer Cells Early in Apoptosis Allows for an Antigen-Driven Antibody Response Elicited by Tumor-Infiltrating B Cells

Hansen

Nielsen

Ditzel

2002

The Journal of Immunology

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“…Two studies in mouse models have attempted to directly evaluate this relationship, but the results led to different conclusions regarding whether the library approach enabled recovery of Abs with the same V H and V L genes sampled by the hybridoma approach (39,40). While direct side-by-side comparisons of hybridomas and combinatorial library studies have not been feasible in humans, studies have documented that repertoire cloning approaches can isolate human Abs with Ag-binding properties identical to those of the donor's serum Abs to infectious pathogens (41,42) and autoAgs (43,44). In the current report, anti-dsDNA Abs were obtained from libraries from two lupus patients, and our sequence analyses and idiotype surveys indicate that these recovered autoAbs can use V regions with predominant H-L combinations that reiterate features of the in vivo response (discussed further below).…”

Section: Discussionmentioning

confidence: 99%

Repertoire cloning of lupus anti-DNA autoantibodies.

Roben¹,

Barbas²,

Sandoval³

et al. 1996

J. Clin. Invest.

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“…Analyses of somatic mutations in mAbs derived from individuals exposed to viruses suggest that the processes of somatic mutation and Ag-affinity selection can play a major role in shaping the Ab response to viruses, including HIV-1 (58,59,(71)(72)(73). The present study directly demonstrates that GCs are a major site for somatic mutation and selection of antiviral Abs in primates.…”

Section: Discussionmentioning

confidence: 99%

Germinal Center Function in the Spleen during Simian HIV Infection in Rhesus Monkeys

Margolin

Saunders

Bronfin

et al. 2006

The Journal of Immunology

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Infection with HIV-1, SIV, or simian HIV is associated with abnormalities in the number, size, and structure of germinal centers (GCs). To determine whether these histopathologic abnormalities are associated with abnormalities in Ab development, we analyzed nucleotide sequences of Igs from splenic GCs of simian HIV-infected macaques. Virus-specific GCs were identified in frozen splenic tissue sections by inverse immunohistochemistry using rHIV-1 gp120 as a probe. B cells from envelope-specific GCs were isolated from these sections using laser capture microdissection. Their Igs were amplified from cDNA using nested PCR, then cloned and sequenced. Nucleotide sequences were recovered from nine multimember clonal lineages. Within each lineage, sequences had similar V-D-J or V-J junctions but differed by somatic mutations distributed throughout the variable domain. The clones were highly mutated, similar to that previously reported for HIV-1-specific human IgG Abs. The average clone had 37 mutations in the V region, for a frequency of 0.11 mutations/base. The mutational pattern was strikingly nonrandom, with somatic mutations occurring preferentially at RGYW/WRCY hotspots. Transition mutations were favored over transversions, with C→T and G→A replacements together accounting for almost one-third of all mutations. Analysis of replacement and silent mutations in the framework and CDRs suggests that the Igs were subjected to affinity selection. These data demonstrate that the process of Ab maturation is not seriously disrupted in GCs during the early stages of immunodeficiency virus infection, and that Env-specific Igs developing in GCs are subject to extensive somatic mutation and profound selection pressures.

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Molecular Profile of an Antibody Response to HIV-1 as Probed by Combinatorial Libraries

Cited by 221 publications

References 0 publications

Translocation of an Intracellular Antigen to the Surface of Medullary Breast Cancer Cells Early in Apoptosis Allows for an Antigen-Driven Antibody Response Elicited by Tumor-Infiltrating B Cells

Translocation of an Intracellular Antigen to the Surface of Medullary Breast Cancer Cells Early in Apoptosis Allows for an Antigen-Driven Antibody Response Elicited by Tumor-Infiltrating B Cells

Repertoire cloning of lupus anti-DNA autoantibodies.

Germinal Center Function in the Spleen during Simian HIV Infection in Rhesus Monkeys

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