Abstract
To increase the inherently weak immunogenicity of synthetic peptide vaccines, we used recombinant DNA techniques to generate chimeras between immunogenic determinants of human immunodeficiency virus type 1 (HIV-1) gp120 and antibody Fab fragments reactive with surface structures displayed specifically on human antigen-presenting cells (APCs), including surface immunoglobulin D (sIgD) and class II major histocompatibility complex (MHC) molecules. Hybridomas producing anti-human MHC class II (HLA-DR) or surface immunoglobulin D monoclonal antibodies (MAbs) that recognize nonpolymorphic determinants were used to clone chimeric Fab gene fragments by employing an established procedure to generate antigen-binding Fab libraries in phagemid vector pComb3. Molecular and immunochemical analysis indicated that the expected chimeric Fab fragments expressing the HIV-1 epitopes were correctly cloned and expressed in Escherichia coli and retained the binding specificity of the native (hybridoma-derived) MAb. The chimeric Fab fragments targeted the linked HIV-1-derived antigenic determinants to the surface of human APCs in vitro, as evidenced by fluorescence-activated cell sorter analysis. Furthermore, such recombinant immunotargeted HIV-1 peptide antigens demonstrated improved immunogenicity over equivalent nonimmunotargeted control antigens, as shown by their ability to stimulate interleukin-2 production by CD4+ T-helper cells from human donors exposed to HIV-1 antigens. These data suggest that immunotargeting of recombinant peptide antigens via the attached Fab fragments facilitates uptake by human APCs with subsequent access to the MHC class II processing pathway, thereby validating the immunotargeting concept for such recombinant subunit vaccines in an in vitro human system.
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- Adelman N. E., Watling D. L., McDevitt H. O. Treatment of (NZB x NZW)F1 disease with anti-I-A monoclonal antibodies. J Exp Med. 1983 Oct 1;158(4):1350–1355. doi: 10.1084/jem.158.4.1350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Agy M. B., Frumkin L. R., Corey L., Coombs R. W., Wolinsky S. M., Koehler J., Morton W. R., Katze M. G. Infection of Macaca nemestrina by human immunodeficiency virus type-1. Science. 1992 Jul 3;257(5066):103–106. doi: 10.1126/science.1621083. [DOI] [PubMed] [Google Scholar]
- Altman A., Dixon F. J. Immunomodifiers in vaccines. Adv Vet Sci Comp Med. 1989;33:301–343. doi: 10.1016/b978-0-12-039233-9.50013-5. [DOI] [PubMed] [Google Scholar]
- Arnon R., Horwitz R. J. Synthetic peptides as vaccines. Curr Opin Immunol. 1992 Aug;4(4):449–453. doi: 10.1016/s0952-7915(06)80037-3. [DOI] [PubMed] [Google Scholar]
- Barbas C. F., 3rd, Kang A. S., Lerner R. A., Benkovic S. J. Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7978–7982. doi: 10.1073/pnas.88.18.7978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berman P. W., Matthews T. J., Riddle L., Champe M., Hobbs M. R., Nakamura G. R., Mercer J., Eastman D. J., Lucas C., Langlois A. J. Neutralization of multiple laboratory and clinical isolates of human immunodeficiency virus type 1 (HIV-1) by antisera raised against gp120 from the MN isolate of HIV-1. J Virol. 1992 Jul;66(7):4464–4469. doi: 10.1128/jvi.66.7.4464-4469.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berzofsky J. A., Bensussan A., Cease K. B., Bourge J. F., Cheynier R., Lurhuma Z., Salaün J. J., Gallo R. C., Shearer G. M., Zagury D. Antigenic peptides recognized by T lymphocytes from AIDS viral envelope-immune humans. Nature. 1988 Aug 25;334(6184):706–708. doi: 10.1038/334706a0. [DOI] [PubMed] [Google Scholar]
- Berzofsky J. A. Development of artificial vaccines against HIV using defined epitopes. FASEB J. 1991 Jul;5(10):2412–2418. doi: 10.1096/fasebj.5.10.1712327. [DOI] [PubMed] [Google Scholar]
- Bolognesi D. P. AIDS. Do antibodies enhance the infection of cells by HIV? Nature. 1989 Aug 10;340(6233):431–432. doi: 10.1038/340431a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bolognesi D. P. Progress in vaccines against AIDS. Science. 1989 Dec 8;246(4935):1233–1234. doi: 10.1126/science.2555922. [DOI] [PubMed] [Google Scholar]
- Carayanniotis G., Barber B. H. Adjuvant-free IgG responses induced with antigen coupled to antibodies against class II MHC. Nature. 1987 May 7;327(6117):59–61. doi: 10.1038/327059a0. [DOI] [PubMed] [Google Scholar]
- Carayanniotis G., Skea D. L., Luscher M. A., Barber B. H. Adjuvant-independent immunization by immunotargeting antigens to MHC and non-MHC determinants in vivo. Mol Immunol. 1991 Mar;28(3):261–267. doi: 10.1016/0161-5890(91)90072-r. [DOI] [PubMed] [Google Scholar]
- Casten L. A., Kaumaya P., Pierce S. K. Enhanced T cell responses to antigenic peptides targeted to B cell surface Ig, Ia, or class I molecules. J Exp Med. 1988 Jul 1;168(1):171–180. doi: 10.1084/jem.168.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Casten L. A., Pierce S. K. Receptor-mediated B cell antigen processing. Increased antigenicity of a globular protein covalently coupled to antibodies specific for B cell surface structures. J Immunol. 1988 Jan 15;140(2):404–410. [PubMed] [Google Scholar]
- Cease K. B., Margalit H., Cornette J. L., Putney S. D., Robey W. G., Ouyang C., Streicher H. Z., Fischinger P. J., Gallo R. C., DeLisi C. Helper T-cell antigenic site identification in the acquired immunodeficiency syndrome virus gp120 envelope protein and induction of immunity in mice to the native protein using a 16-residue synthetic peptide. Proc Natl Acad Sci U S A. 1987 Jun;84(12):4249–4253. doi: 10.1073/pnas.84.12.4249. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cease K. B. Peptide component vaccine engineering: targeting the AIDS virus. Int Rev Immunol. 1990;7(1):85–107. doi: 10.3109/08830189009061767. [DOI] [PubMed] [Google Scholar]
- Chesebro B., Wehrly K. Development of a sensitive quantitative focal assay for human immunodeficiency virus infectivity. J Virol. 1988 Oct;62(10):3779–3788. doi: 10.1128/jvi.62.10.3779-3788.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chesnut R. W., Grey H. M. Studies on the capacity of B cells to serve as antigen-presenting cells. J Immunol. 1981 Mar;126(3):1075–1079. [PubMed] [Google Scholar]
- Clerici M., Berzofsky J. A., Shearer G. M., Tacket C. O. Exposure to human immunodeficiency virus (HIV) type I indicated by HIV-specific T helper cell responses before detection of infection by polymerase chain reaction and serum antibodies [corrected]. J Infect Dis. 1991 Jul;164(1):178–182. doi: 10.1093/infdis/164.1.178. [DOI] [PubMed] [Google Scholar]
- Clerici M., Giorgi J. V., Chou C. C., Gudeman V. K., Zack J. A., Gupta P., Ho H. N., Nishanian P. G., Berzofsky J. A., Shearer G. M. Cell-mediated immune response to human immunodeficiency virus (HIV) type 1 in seronegative homosexual men with recent sexual exposure to HIV-1. J Infect Dis. 1992 Jun;165(6):1012–1019. doi: 10.1093/infdis/165.6.1012. [DOI] [PubMed] [Google Scholar]
- Clerici M., Shearer G. M. A TH1-->TH2 switch is a critical step in the etiology of HIV infection. Immunol Today. 1993 Mar;14(3):107–111. doi: 10.1016/0167-5699(93)90208-3. [DOI] [PubMed] [Google Scholar]
- Clerici M., Stocks N. I., Zajac R. A., Boswell R. N., Bernstein D. C., Mann D. L., Shearer G. M., Berzofsky J. A. Interleukin-2 production used to detect antigenic peptide recognition by T-helper lymphocytes from asymptomatic HIV-seropositive individuals. Nature. 1989 Jun 1;339(6223):383–385. doi: 10.1038/339383a0. [DOI] [PubMed] [Google Scholar]
- Clerici M., Tacket C. O., Via C. S., Lucey D. R., Muluk S. C., Zajac R. A., Boswell R. N., Berzofsky J. A., Shearer G. M. Immunization with subunit human immunodeficiency virus vaccine generates stronger T helper cell immunity than natural infection. Eur J Immunol. 1991 Jun;21(6):1345–1349. doi: 10.1002/eji.1830210603. [DOI] [PubMed] [Google Scholar]
- Cohen J. AIDS research shifts to immunity. Science. 1992 Jul 10;257(5067):152–154. doi: 10.1126/science.1631539. [DOI] [PubMed] [Google Scholar]
- Daar E. S., Moudgil T., Meyer R. D., Ho D. D. Transient high levels of viremia in patients with primary human immunodeficiency virus type 1 infection. N Engl J Med. 1991 Apr 4;324(14):961–964. doi: 10.1056/NEJM199104043241405. [DOI] [PubMed] [Google Scholar]
- Day M. J., Tse A. G., Puklavec M., Simmonds S. J., Mason D. W. Targeting autoantigen to B cells prevents the induction of a cell-mediated autoimmune disease in rats. J Exp Med. 1992 Mar 1;175(3):655–659. doi: 10.1084/jem.175.3.655. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Demotz S., Grey H. M., Sette A. The minimal number of class II MHC-antigen complexes needed for T cell activation. Science. 1990 Aug 31;249(4972):1028–1030. doi: 10.1126/science.2118680. [DOI] [PubMed] [Google Scholar]
- Desrosiers R. C. Inching towards a vaccine for AIDS. Curr Biol. 1992 Mar;2(3):162–163. doi: 10.1016/0960-9822(92)90278-i. [DOI] [PubMed] [Google Scholar]
- Dreher M. L., Gherardi E., Skerra A., Milstein C. Colony assays for antibody fragments expressed in bacteria. J Immunol Methods. 1991 Jun 3;139(2):197–205. doi: 10.1016/0022-1759(91)90189-m. [DOI] [PubMed] [Google Scholar]
- Eynon E. E., Parker D. C. Small B cells as antigen-presenting cells in the induction of tolerance to soluble protein antigens. J Exp Med. 1992 Jan 1;175(1):131–138. doi: 10.1084/jem.175.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Finkelman F. D., Snapper C. M., Mountz J. D., Katona I. M. Polyclonal activation of the murine immune system by a goat antibody to mouse IgD. IX. Induction of a polyclonal IgE response. J Immunol. 1987 May 1;138(9):2826–2830. [PubMed] [Google Scholar]
- Golding H., Robey F. A., Gates F. T., 3rd, Linder W., Beining P. R., Hoffman T., Golding B. Identification of homologous regions in human immunodeficiency virus I gp41 and human MHC class II beta 1 domain. I. Monoclonal antibodies against the gp41-derived peptide and patients' sera react with native HLA class II antigens, suggesting a role for autoimmunity in the pathogenesis of acquired immune deficiency syndrome. J Exp Med. 1988 Mar 1;167(3):914–923. doi: 10.1084/jem.167.3.914. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goroff D. K., Stall A., Mond J. J., Finkelman F. D. In vitro and in vivo B lymphocyte-activating properties of monoclonal anti-delta antibodies. I. Determinants of B lymphocyte-activating properties. J Immunol. 1986 Apr 1;136(7):2382–2392. [PubMed] [Google Scholar]
- Hosmalin A., Nara P. L., Zweig M., Lerche N. W., Cease K. B., Gard E. A., Markham P. D., Putney S. D., Daniel M. D., Desrosiers R. C. Priming with T helper cell epitope peptides enhances the antibody response to the envelope glycoprotein of HIV-1 in primates. J Immunol. 1991 Mar 1;146(5):1667–1673. [PubMed] [Google Scholar]
- Javaherian K., Langlois A. J., McDanal C., Ross K. L., Eckler L. I., Jellis C. L., Profy A. T., Rusche J. R., Bolognesi D. P., Putney S. D. Principal neutralizing domain of the human immunodeficiency virus type 1 envelope protein. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6768–6772. doi: 10.1073/pnas.86.17.6768. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kawamura H., Berzofsky J. A. Enhancement of antigenic potency in vitro and immunogenicity in vivo by coupling the antigen to anti-immunoglobulin. J Immunol. 1986 Jan;136(1):58–65. [PubMed] [Google Scholar]
- Koff W. C., Glass M. J. Future directions in HIV vaccine development. AIDS Res Hum Retroviruses. 1992 Aug;8(8):1313–1315. doi: 10.1089/aid.1992.8.1313. [DOI] [PubMed] [Google Scholar]
- LaRosa G. J., Davide J. P., Weinhold K., Waterbury J. A., Profy A. T., Lewis J. A., Langlois A. J., Dreesman G. R., Boswell R. N., Shadduck P. Conserved sequence and structural elements in the HIV-1 principal neutralizing determinant. Science. 1990 Aug 24;249(4971):932–935. doi: 10.1126/science.2392685. [DOI] [PubMed] [Google Scholar]
- Lanzavecchia A. Receptor-mediated antigen uptake and its effect on antigen presentation to class II-restricted T lymphocytes. Annu Rev Immunol. 1990;8:773–793. doi: 10.1146/annurev.iy.08.040190.004013. [DOI] [PubMed] [Google Scholar]
- Lasarte J. J., Sarobe P., Gullón A., Prieto J., Borrás-Cuesta F. Induction of cytotoxic T lymphocytes in mice against the principal neutralizing domain of HIV-1 by immunization with an engineered T-cytotoxic-T-helper synthetic peptide construct. Cell Immunol. 1992 Apr 15;141(1):211–218. doi: 10.1016/0008-8749(92)90140-k. [DOI] [PubMed] [Google Scholar]
- Lees A., Morris S. C., Thyphronitis G., Holmes J. M., Inman J. K., Finkelman F. D. Rapid stimulation of large specific antibody responses with conjugates of antigen and anti-IgD antibody. J Immunol. 1990 Dec 1;145(11):3594–3600. [PubMed] [Google Scholar]
- Myers C. D. Role of B cell antigen processing and presentation in the humoral immune response. FASEB J. 1991 Aug;5(11):2547–2553. doi: 10.1096/fasebj.5.11.1907935. [DOI] [PubMed] [Google Scholar]
- Palker T. J., Matthews T. J., Langlois A., Tanner M. E., Martin M. E., Scearce R. M., Kim J. E., Berzofsky J. A., Bolognesi D. P., Haynes B. F. Polyvalent human immunodeficiency virus synthetic immunogen comprised of envelope gp120 T helper cell sites and B cell neutralization epitopes. J Immunol. 1989 May 15;142(10):3612–3619. [PubMed] [Google Scholar]
- Parker D. C., Eynon E. E. Antigen presentation in acquired immunological tolerance. FASEB J. 1991 Oct;5(13):2777–2784. doi: 10.1096/fasebj.5.13.1916102. [DOI] [PubMed] [Google Scholar]
- Pierce S. K., Morris J. F., Grusby M. J., Kaumaya P., van Buskirk A., Srinivasan M., Crump B., Smolenski L. A. Antigen-presenting function of B lymphocytes. Immunol Rev. 1988 Dec;106:149–180. doi: 10.1111/j.1600-065x.1988.tb00778.x. [DOI] [PubMed] [Google Scholar]
- Redfield R. R., Birx D. L., Ketter N., Tramont E., Polonis V., Davis C., Brundage J. F., Smith G., Johnson S., Fowler A. A phase I evaluation of the safety and immunogenicity of vaccination with recombinant gp160 in patients with early human immunodeficiency virus infection. Military Medical Consortium for Applied Retroviral Research. N Engl J Med. 1991 Jun 13;324(24):1677–1684. doi: 10.1056/NEJM199106133242401. [DOI] [PubMed] [Google Scholar]
- Rock K. L., Benacerraf B., Abbas A. K. Antigen presentation by hapten-specific B lymphocytes. I. Role of surface immunoglobulin receptors. J Exp Med. 1984 Oct 1;160(4):1102–1113. doi: 10.1084/jem.160.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Snider D. P., Segal D. M. Efficiency of antigen presentation after antigen targeting to surface IgD, IgM, MHC, Fc gamma RII, and B220 molecules on murine splenic B cells. J Immunol. 1989 Jul 1;143(1):59–65. [PubMed] [Google Scholar]
- Sternberg S. HIV comes in five family groups. Science. 1992 May 15;256(5059):966–966. doi: 10.1126/science.1589779. [DOI] [PubMed] [Google Scholar]
- Stevens V. C. Future perspectives for vaccine development. Scand J Immunol Suppl. 1992;11:137–143. doi: 10.1111/j.1365-3083.1992.tb01637.x. [DOI] [PubMed] [Google Scholar]
- Takahashi H., Cohen J., Hosmalin A., Cease K. B., Houghten R., Cornette J. L., DeLisi C., Moss B., Germain R. N., Berzofsky J. A. An immunodominant epitope of the human immunodeficiency virus envelope glycoprotein gp160 recognized by class I major histocompatibility complex molecule-restricted murine cytotoxic T lymphocytes. Proc Natl Acad Sci U S A. 1988 May;85(9):3105–3109. doi: 10.1073/pnas.85.9.3105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Takahashi H., Germain R. N., Moss B., Berzofsky J. A. An immunodominant class I-restricted cytotoxic T lymphocyte determinant of human immunodeficiency virus type 1 induces CD4 class II-restricted help for itself. J Exp Med. 1990 Feb 1;171(2):571–576. doi: 10.1084/jem.171.2.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tony H. P., Parker D. C. Major histocompatibility complex-restricted, polyclonal B cell responses resulting from helper T cell recognition of antiimmunoglobulin presented by small B lymphocytes. J Exp Med. 1985 Jan 1;161(1):223–241. doi: 10.1084/jem.161.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wigzell H. Prospects for an HIV-vaccine. FASEB J. 1991 Jul;5(10):2406–2411. doi: 10.1096/fasebj.5.10.2065889. [DOI] [PubMed] [Google Scholar]
- Wigzell H. Prospects for an HIV-vaccine. FASEB J. 1991 Jul;5(10):2406–2411. doi: 10.1096/fasebj.5.10.2065889. [DOI] [PubMed] [Google Scholar]
- Zaghouani H., Kuzo Y., Kuzo H., Mann N., Daian C., Bona C. Engineered immunoglobulin molecules as vehicles for T cell epitopes. Int Rev Immunol. 1993;10(2-3):265–278. doi: 10.3109/08830189309061701. [DOI] [PubMed] [Google Scholar]