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Specific Killing of HIV-infected Lymphocytes by a Recombinant Immunotoxin Directed against the HIV-1 Envelope Glycoprotein

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Abstract

Background

3B3 is a high-affinity anti-gp120 antibody that neutralizes a wide range of primary and laboratory isolates of HIV-1. The parental antibody was isolated from a combinatorial phage display library constructed from bone marrow RNA of an HIV-infected individual. We have generated a highly active immunotoxin using the 3B3 single-chain Fv (scFv) which can specifically kill lymphocytes infected by HIV-1.

Materials and Methods

We used recombinant DNA technology to clone the Fv fragment of 3B3 and produce a single-chain Fv (scFv). 3B3 scFv was then fused to a truncated version of Pseudomonas exotoxin A (PE38), giving rise to a recombinant immunotoxin 3B3(Fv)-PE38 that was expressed in E. coli and purified to near homogeneity.

Results

3B3(Fv)-PE38 binds with the same affinity as the parental Fab antibody to the MN strain of gp120. The immunotoxin specifically kills a gp120-expressing transfected cell line and a chronically HIV-infected lymphocytic cell line. The immunotoxin is very stable at 37°C, retaining 80% of its original activity after 24 hr.

Conclusions

Potent immunotoxins such as 3B3(Fv)-PE38 could be utilized in combination with multidrug cocktails that limit viral replication to help reduce viral reservoirs in patients with AIDS.

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Acknowledgments

We thank R. Beers for performing the BioCore studies, members of Dr. Pastan’s laboratory for critical reading of the manuscript, V. Fogg for cell culture assistance, and R. Mann and J. Evans for editorial assistance. C. F. B. was supported by NIH grant no. AI37470.

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Authors and Affiliations

  1. Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Building 37, Room 4E16, 37 Convent Drive, MSC 4255, Bethesda, Maryland, 20892-4255, USA

    Tapan K. Bera &  Ira Pastan

  2. Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Paul E. Kennedy & Edward A. Berger

  3. The Skaggs Institute for Chemical Biology, Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, USA

    Carlos F. Barbas III

Authors
  1. Tapan K. Bera
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  2. Paul E. Kennedy
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  3. Edward A. Berger
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  4. Carlos F. Barbas III
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  5. Ira Pastan
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Communicated by I. Pastan.

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Bera, T.K., Kennedy, P.E., Berger, E.A. et al. Specific Killing of HIV-infected Lymphocytes by a Recombinant Immunotoxin Directed against the HIV-1 Envelope Glycoprotein. Mol Med 4, 384–391 (1998). https://doi.org/10.1007/BF03401745

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  • DOI: https://doi.org/10.1007/BF03401745

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