Chimeric HIV-1 Envelope Glycoproteins with Potent Intrinsic Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) Activity*

Isik, Gözde; Montfort, Thijs van; Boot, Maikel; Jiménez, Viviana Cobos; Kootstra, Neeltje A.; Sanders, Rogier W.

doi:10.1371/journal.pone.0060126

Cited by 7 publications

(16 citation statements)

References 63 publications

(22 reference statements)

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“…Yan, K. Hu, and Q. Hu, unpublished observations). In addition to the linker, the construction order of the fusion proteins may affect protein activity (34,54). We demonstrated that fusion constructs with CCL19 at the N-or C-terminal end had comparable abilities to enhance immune responses.…”

Section: Discussionmentioning

confidence: 99%

“…It is generally accepted that Ags fused to ligands of target cell surface molecules can facilitate Ag uptake and presentation and, consequently, induce the activation of responsive immunocytes (33)(34)(35)(36)(37). Fusion of HIV-1 Env to several cytokines, including APRIL, BAFF, CD40L, IL-12, and GM-CSF, demonstrated improved immunogenicity of the DNA vaccines (33,35,36).…”

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confidence: 99%

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Immunization with HSV-2 gB-CCL19 Fusion Constructs Protects Mice against Lethal Vaginal Challenge

Yan

Deng

et al. 2015

The Journal of Immunology

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There is a lack of an HSV-2 vaccine, in part as the result of various factors that limit robust and long-term memory immune responses at the mucosal portals of viral entry. We previously demonstrated that chemokine CCL19 augmented mucosal and systemic immune responses to HIV-1 envelope glycoprotein. Whether such enhanced immunity can protect animals against virus infection remains to be addressed. We hypothesized that using CCL19 in a fusion form to direct an immunogen to responsive immunocytes might have an advantage over CCL19 being used in combination with an immunogen. We designed two fusion constructs, plasmid (p)gBIZCCL19 and pCCL19IZgB, by fusing CCL19 to the C- or N-terminal end of the extracellular HSV-2 glycoprotein B (gB) with a linker containing two (Gly4Ser)2 repeats and a GCN4-based isoleucine zipper motif for self-oligomerization. Following immunization in mice, pgBIZCCL19 and pCCL19IZgB induced strong gB-specific IgG and IgA in sera and vaginal fluids. The enhanced systemic and mucosal Abs showed increased neutralizing activity against HSV-2 in vitro. Measurement of gB-specific cytokines demonstrated that gB-CCL19 fusion constructs induced balanced Th1 and Th2 cellular immune responses. Moreover, mice vaccinated with fusion constructs were well protected from intravaginal lethal challenge with HSV-2. Compared with pgB and pCCL19 coimmunization, fusion constructs increased mucosal surface IgA+ cells, as well as CCL19-responsive immunocytes in spleen and mesenteric lymph nodes. Our findings indicate that enhanced humoral and cellular immune responses can be achieved by immunization with an immunogen fused to a chemokine, providing information for the design of vaccines against mucosal infection by HSV-2 and other sexually transmitted viruses.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Immunization with HSV-2 gB-CCL19 Fusion Constructs Protects Mice against Lethal Vaginal Challenge

Yan

Deng

et al. 2015

The Journal of Immunology

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“…Reactivity of Env proteins with MAbs or receptor mimics were measured by anti-trimer ELISA as described previously using Ni-NTA HisSorb 96-well plates (Qiagen, Venlo, The Netherlands)(10,34–36,47). Equal input levels of proteins were verified by SDS-PAGE followed by western blot analysis.…”

Section: Methodsmentioning

confidence: 99%

“…The anti-cytokine ELISA was performed as previously described for the anti-trimer ELISA (10,34–36,47). The wells were coated with supernatants containing His-tagged cytokines (Supplemental Table I) or mock media (or Env wt for comparative purposes) and the plates were washed twice with Tris-buffered saline (TBS).…”

Section: Methodsmentioning

confidence: 99%

“…We have shown that fusing Env trimers to co-stimulatory molecules such as CD40L and APRIL that facilitate B cell help can be beneficial to Env immunogenicity (5,6). Furthermore, replacing Env variable loops 1 and 2 (V1V2) with GM-CSF increased both anti-gp120 binding Ab titers and T-cell responses in mice (34,35), while replacing the same loops with IL-21 enhanced Ig secretion and the formation of human plasmablast-like B cells in vitro (36). Both GM-CSF and IL-21 cytokines have a good safety profile in clinical studies.…”

Section: Introductionmentioning

confidence: 99%

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Autoantibodies Induced by Chimeric Cytokine-HIV Envelope Glycoprotein Immunogens

Isik

Montfort

Chung

et al. 2014

The Journal of Immunology

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Cytokines are often used as adjuvants to increase the immunogenicity of vaccines as they can improve the immune response and/or direct it into a desired direction. As an alternative to co-delivering antigens and cytokines separately they can be fused into a composite protein, with the advantage that both moieties act on the same immune cells. The HIV-1 envelope glycoprotein (Env) spike, located on the outside of virus particles and the only relevant protein for the induction of neutralizing antibodies (NAbs), is poorly immunogenic. The induction of anti-Env Abs can be improved by coupling Env proteins to co-stimulatory molecules such as a proliferation inducing ligand (APRIL). Here, we evaluated the immunogenicity of chimeric molecules containing uncleaved Env gp140 fused to the species-matched cytokines IL-21 or GM-CSF in rabbits and mice. Each cytokine was either fused to the C-terminus of Env or embedded within Env at the position of the variable loops 1 and 2 (V1V2). The cytokine components of the chimeric Env-GM-CSF and Env-IL-21 molecules were functional in vitro, but none of the Env-cytokine fusion proteins resulted in improved Ab responses in vivo. Both the Env-GM-CSF and the Env-IL-21 molecules induced strong anti-cytokine Ab responses, in both test species. These autoimmune responses were independent of the location of the cytokine in the chimeric Env molecules; in that they were induced by cytokines inserted within the V1V2 of Env or fused to its Ct. The induction of undesired autoimmune responses should be considered when using cytokines as co-stimulatory molecules in fusion proteins.

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Development of broadly neutralizing antibodies from autologous neutralizing antibody responses in HIV infection

Derdeyn

Moore

Morris

2014

Current Opinion in HIV and AIDS

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Purpose of Review Detailed genetic and structural characterization has revealed that broadly neutralizing antibodies (bnAbs) against HIV-1 have unusually high levels of somatic hypermutation, long CDRH3 domains, and the ability to target one of four sites of vulnerability on the HIV-1 envelope (Env) glycoproteins. A current priority is to understand how bnAbs are generated during natural infection, and translate this information into immunogens that can elicit bnAb following vaccination. Recent Findings Strain-specific neutralizing antibodies (nAb) can acquire broad neutralizing capacity when the transmitted/founder Env or a specific Env variant is recognized by an unmutated rearranged germline that has the capacity to develop bnAb like features. This could be a relatively infrequent event, as only certain germlines appear to possess inherent features needed for bnAb activity. Furthermore, the glycosylation pattern and diversity of circulating HIV-1 Envs, as well as the state of the B cell compartment, may influence the activation and maturation of certain antibody lineages. Summary Collectively, studies over the last year suggest that the development of HIV-1 Env immunogens that bind and activate bnAb-like germlines is feasible. However, more information about the features of Env variants and the host factors that lead to breadth during natural infection is needed to elicit bnAbs through immunization.

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Chimeric HIV-1 Envelope Glycoproteins with Potent Intrinsic Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) Activity*

Cited by 7 publications

References 63 publications

Immunization with HSV-2 gB-CCL19 Fusion Constructs Protects Mice against Lethal Vaginal Challenge

Immunization with HSV-2 gB-CCL19 Fusion Constructs Protects Mice against Lethal Vaginal Challenge

Autoantibodies Induced by Chimeric Cytokine-HIV Envelope Glycoprotein Immunogens

Development of broadly neutralizing antibodies from autologous neutralizing antibody responses in HIV infection

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